Nerve and tumor homing agents and methods of use thereof

Abstract

Disclosed herein a method of visualizing and treating a cancer in a subject in need thereof, the method comprising: administering to the subject a tumor homing agent and a nerve homing agent. The technology described herein provides direct visualization of tissue, cells, and margins and direct visualization of nerves simultaneously in surgical settings and pathology samples in order to improve surgical outcomes in cancer surgeries and reduce comorbidities for patients.

Description

NERVE AND TUMOR HOMING AGENTS AND METHODS OF USE THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit of priority under 35 U. S. C. § 119(e) of U. S. Provisional Application No. 63 / 718,543, filed Nov. 8, 2024. The disclosure of the prior applications is considered part of and is herein incorporated by reference in the disclosure of this application in its entirety.SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in extensible Markup Language (XML) format and is hereby incorporated by reference in its entirety. Said XML copy, created on November 4, 2025, is named “116772-1528696-726WOI SL.xml” and is 553.620 bytes in size.BACKGROUND

[0003] For many cancers and solid tumors, surgery is a primary treatment option, during which surgeons face two main challenges or goals during a surgery to ensure post-surgical quality of life: 1) complete cancer resection and 2) preservation of normal tissue structures. Unfortunately, these goals are not necessarily synergistic or complementary using today’s standard of care surgical methodologies in many cancers. Thus, there is a need in the art for improved methods for cancer identification and treatment while identifying and preserving normal tissues.SUMMARY

[0004] In various aspects, the present disclosure provides a method of detecting a tissue or cell and a nerve tissue or cell in a subject in need thereof, the method comprising administering to a subject a tumor homing agent and a nerve homing agent.

[0005] In some aspects, the detecting comprises fluorescence imaging. In some aspects, the tumor homing agent is fluorescent, the nerve homing agent is fluorescent, or both.

[0006] In some aspects, the nerve homing agent and the tumor homing agent have a minimum spectral gap in emission wavelength of at least 5 nm to at least 800 nm. In some aspects, the minimum spectral gap in emission wavelength is at least 5 nm, at least 10 nm, at least 20 nm, at least 30 nm, at least 40 nm, at least 50 nm, at least 60 nm, at least 70 nm, at least 80 nm, at least 90 nm, at least 100 nm, at least 110 nm, at least 120 nm, at least 130 nm, at least 140 nm, at least 150 nm, at least 160 nm. at least 170 nm, at least 180 nm. at least 190 nm, or at least 200 nm. In some aspects, an emission wavelength spectrum of the nerve homing agent overlaps an excitation wavelength spectrum of the tumor homing agent by at least 0.5%, at least 1%, at least2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, or at least 40%.

[0007] In some aspects, the nerve homing agent and tumor homing agent have a spectral gap in peak excitation wavelength of at least 5 nm to at least 800 nm. In some aspects, the spectral gap in peak excitation wavelength of the nerve homing agent relative to the tumor homing agent is at least 5 nm, at least 10 nm, at least 20 nm, at least 30 nm, at least 40 nm, at least 50 nm, at least 60 nm, at least 70 nm, at least 80 nm, at least 90 nm, at least 100 nm, at least 110 nm, at least 120 nm, at least 130 nm, at least 140 nm, at least 150 nm, at least 160 nm, at least 170 nm, at least 180 nm, at least 190 nm, or at least 200 nm.

[0008] In some aspects, an excitation wavelength spectrum of the nerve homing agent does not overlap with an emission wavelength spectrum of the tumor homing agent. In some aspects, an excitation wavelength spectrum of the nerve homing agent completely or partially overlaps an emission wavelength spectrum of the tumor homing agent. In some aspects, an emission wavelength spectrum of the nerve homing agent does not overlap an excitation wavelength spectrum of the tumor homing agent. In some aspects, an emission wavelength spectrum of the nerve homing agent completely or partially overlaps an excitation wavelength spectrum of the tumor homing agent.

[0009] In some aspects, the tumor homing agent has a peak emission between 600 to 900 nm. In some aspects, the tumor homing agent is excited, imaged, detected, or any combination thereof, at 715 to 800 nm excitation. In some aspects, the tumor homing agent is detected at 815 to 875 nm emission. In some aspects, the tumor homing agent detects a physiologic structure or cell between at least 10 pm and 25 pm in diameter. In some aspects, the nerve homing agent has a peak emission between 600-775 nm inclusive.

[0010] In some aspects, the method further comprises performing a fluorescence guided surgery (FGS) procedure on the subject in need thereof. In some aspects, the fluorescence guided surgery (FGS) is performed with the assistance of an imaging system. In some aspects, the imaging system is selected from the group consisting of surgical microscope, confocal microscope, fluorescence scope, exoscope, endoscope, ophthalmoscope, retinal camera system, optical coherence tomography (OCT) system, surgical robot, and Quest Spectrum imagingsystem, Fluobeam devices, Lab Flare imaging system, or Solaris imaging system, or other imaging system equipped with two-color imaging capabilities.

[0011] In some aspects, the nerve homing agent is retained by, or accumulates in and / or binds to a nerve tissue or nerve cell. In some aspects, the method further comprises detecting the presence or absence of the nerve homing agent in a tissue or cell, wherein the presence of the nerve homing agent in the tissue or cell indicates the presence of nerve tissue or a nerve cell. In some aspects, the tumor homing agent is retained by, or accumulates in and / or binds to cancerous tissue or a cancer cell, or a vascular lesion. In some aspects, the method further comprises detecting the presence or absence of the tumor homing agent in a tissue or cell, wherein the presence of the tumor homing agent in the tissue or cell indicates the presence of a cancerous tissue or a cancer cell, or a vascular lesion.

[0012] In some aspects, the method further comprises treating a cancer or a vascular lesion in the subject in need thereof. In some aspects, the treating further comprises a surgery for surgically removing a cancerous tumor, a cancerous tissue, or a cancerous cell, or vascular lesion, from the subject in need thereof by performing a surgery. In some aspects, surgically removing the cancerous tumor, the cancerous tissue, or the cancerous cell, or vascular lesion, from the subject in need thereof spares the nerve tissue or nerve cell, avoids damaging the nerve tissue or nerve cell, or does not damage a nerve tissue or nerve cell in the subject in need thereof. In some aspects, surgically removing the cancerous tumor, the cancerous tissue, or the cancerous cell, or vascular lesion, from the subject in need thereof shows sparing the nerve tissue or nerve cell, avoiding damaging the nerve tissue or nerve cell, or not damaging a nerve tissue or nerve cell ex vivo in pathology samples obtained from the subject in need thereof or in situ in the patient in need thereof. In some aspects, the surgery is fluorescence guided surgery (FGS).

[0013] In some aspects, the cancer is glioma, astrocytoma, medulloblastoma, choroids plexus carcinoma, ependymoma, neuroblastoma, vestibular schwannoma, carcinoma, meningioma, head and neck cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, breast cancer, intestinal cancer, pancreatic cancer, liver cancer, kidney cancer, sarcoma, osteosarcoma, rhabdomyosarcoma, Ewing’s sarcoma, gastrointestinal stromal tumors, melanoma, ovarian cancer, cervical cancer, lymphoma, thyroid cancer, anal cancer, colorectal cancer, endometrial cancer, laryngeal cancer, multiple myeloma, prostate cancer, retinoblastoma, gastric cancer, testicular cancer, Wilm’s tumor, or any combination thereof, or where the vascular lesion is a cavernous angioma, cavernous hemangioma, or cerebral cavernous malformation (CCM), or any combination thereof. In some aspects, the cancer is a head and neck cancer, a brain cancer, abrain tumor, breast cancer, melanoma, sarcoma, basal cell carcinoma, squamous cell carcinoma, lung cancer, colorectal cancer, prostate cancer, bladder cancer, or any combination thereof. In some aspects, the head and neck cancer is a head and neck squamous cell carcinoma. In some aspects, the breast cancer is invasive ductal carcinoma, ductal carcinoma in situ breast cancer, invasive lobular carcinoma, lobular carcinoma in situ breast cancer, triple negative breast cancer, or any combination thereof.

[0014] In some aspects, the tumor homing agent, the nerve homing agent, or both, are formulated for intravenous administration, intravenous bolus administration, intravenous infusion administration, intramuscular administration, subcutaneous administration, intraperitoneal administration, oral administration, sublingual administration, topical administration, transdermal administration using electroporation, intralesional administration, or a combination thereof. In some aspects, the tumor homing agent and the nerve homing agent are co-administered or are sequentially administered. In some aspects, the tumor homing agent is administered prior to the nerve homing agent, or wherein the nerve homing agent is administered prior to the tumor homing agent, or wherein the tumor homing agent and the nerve homing agent are administered at the same time. In some aspects, the tumor homing agent is administered 1 to 24 hours before or 1 to 3 days before a surgery and the nerve homing agent is administered 10 to 120 minutes before or 2 to 48 hours before a surgery.

[0015] In some aspects, the tumor homing agent comprises a polypeptide comprising: (a) a sequence of any one of SEQ ID NO: 482 - SEQ ID NO: 485, or a fragment thereof, (b) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512, or a fragment thereof, or (c) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 9, or a fragment thereof. In some aspects, the polypeptide comprises SEQ ID NO: 9, or a fragment thereof.

[0016] In some aspects, the fragment of the polypeptide has a length of at least 25 residues. In some aspects, each amino acid of the polypeptide is independently selected as an L- or D-enantiomer. In some aspects, the polypeptide contains no lysine residues. In some aspects, the polypeptide contains a single lysine residue. In some aspects, the single lysine residue is located at a position corresponding to K-27 of native chlorotoxin, K-23 of native chlorotoxin, or K-15 of native chlorotoxin. In some aspects, one, two, or three methionine residues of the polypeptide are replaced with other amino acids. In some aspects, the N-terminus of the polypeptide is blocked by acetylation or cyclization. In some aspects, the polypeptide comprises at least 4, at least 5, or at least 6 disulfide bonds. In some aspects, the polypeptide has a length of no less than21 amino acid residues, no less than 22 amino acid residues, no less than 23 amino acid residues, no less than 24 amino acid residues, no less than 25 amino acid residues, no less than 26 amino acid residues, no less than 27 amino acid residues, no less than 28 amino acid residues, no less than 29 amino acid residues, no less than 30 amino acid residues, no less than 31 amino acid residues, no less than 32 amino acid residues, no less than 33 amino acid residues, no less than 34 amino acid residues, no less than 5 amino acid residues, no less than 36 amino acid residues, no less than 37 amino acid residues, no less than 38 amino acid residues, no less than 39 amino acid residues, or no less than 40 amino acid residues. In some aspects, the polypeptide comprises an isoelectric point at least 8.0, at least 8.5, or at least 9.0.

[0017] In some aspects, the polypeptide is conjugated to an agent. In some aspects, the agent is a detectable agent or a first therapeutic agent. In some aspects, the polypeptide is conjugated to the agent via a cleavable linker or a stable linker. In some aspects, the polypeptide comprises: (a) a single lysine residue and the agent is conjugated to the polypeptide at the single lysine residue, or (b) no lysine residues and the agent is conjugated to the polypeptide at the N-terminus of the polypeptide.

[0018] In some aspects, the tumor homing agent comprises the structure of Formula (I), or a pharmaceutically acceptable salt thereof:R1, R2, R', R4, R5, R6, R7, R8, R1’, and Rlbare each independently selected from hydrogen, Ci-Ce alkyl, Ci-Ce alkylene-COOH, sulfonate, Ci-Ce alkylene-sulfonate, - COOH, -SO2-NH2. or Ci-Ce alkoxy;R9is hydrogen, sulfonate, amine, or -COOH;L1is C3-C6alkylene;L2is C1-C10 alkylene;L3is a bond, –O–, –NR10–, –NR10–C1-C6alkylene–, –O-NR10–, –NR10–C1-C6alkylene–(O-C1-C6alkylene)n–, -NR10-L4-, -NR10-CJ-C6 alkylene-NR11- (C (= O) -Ci-Ce alkylene-O-)m-. or –NR10–C1-C6alkylene—NR10–C1-C6alkylene—NR10–C1-C6alkylene–;L4is a bond, -heterocyclyl-, or -heterocyclyl-Ci-Cg alkylene-;R10is hydrogen or C1-C6alkyl;R11is hydrogen or C1-C6alkyl;R12and R13are independently selected from hydrogen, Ci-Ce alkyl, or R12and R13are joined together along with the other atoms to which they are attached to form a 5- membered or 6-membered carbocyclic or heterocyclic ring;R14is hydrogen or C1-C6alkylene, –(L5)–aryl, –(L5)–aryl–R21, –(L5)–heteroaryl, –(L5)–heteroaryl–R21, –NR17R18, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;L5is a bond, Ci-Cio alkylene, -O-, -NR10-;R17and R18are each independently hydrogen or aryl;R19and R20are independently selected from hydrogen, Ci-Ce alkyl, R14and R19are joined together along with the other atoms to which they are attached to form a 5- membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5 -membered or 6-membered carbocyclic or heterocyclic ring;R21is hydrogen, sulfonate, or -COOH;n is 0, 1, 2, or 3;m is 0, 1, 2, or 3;p is 0, 1, 2, or 3;q is 0, 1, 2, or 3; andA4is a polypeptide.

[0019] In some aspects, R9is sulfonate.

[0020] In some aspects, A4has at least 90% sequence identity with: (a) a sequence of any one of SEQ ID NO: 482 - SEQ ID NO: 485, or a fragment thereof, (b) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512, or a fragment thereof, or (c) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity’ to SEQ ID NO: 9, or a fragment thereof. In some aspects, A4is SEQ ID NO: 9 or a fragment thereof. In some aspects, A4comprises at least 25 amino acid residues.

[0021] In some aspects, R3, R4, R5, R6of the Formula (I), are each independently methyl; R1, R2, R7, R8, R15, and R16are each independently hydrogen; R10is hydrogen; R12, R13, R14, R19, and R20are each independently hydrogen; L1is butylene; and L2is pentylene. In some aspects, L3is attached to A4at a lysine amino acid residue of the polypeptide.

[0022] In some aspects, Formula (I) is conjugated to polyethylene glycol (PEG), hydroxy ethyl starch, polyvinyl alcohol, a water soluble polymer, a zwitterionic water soluble polymer, a water soluble poly(amino acid), an albumin, or a fatty acid.

[0023] In some aspects, Formula (II) further comprises a second therapeutic agent attached to A4, wherein the second therapeutic agent is selected from a radioisotope, toxin, cytotoxic agent, enzyme, sensitizing drug, nucleic acid, interfering RNA, antibody, anti-angiogenic agent, cisplatin, anti-metabolite, mitotic inhibitor, growth factor inhibitor, paclitaxel, temozolomide, topotecan, fluorouracil, vincristine, vinblastine, procarbazine, dacarbazine, altretamine, methotrexate, mercaptopurine, thioguanine, fludarabine phosphate, cladribine, pentostatin, cytarabine, azacitidine, etoposide, teniposide, irinotecan, docetaxel, doxorubicin, daunorubicin, dactinomycin, idarubicin, plicamycin, mitomycin, bleomycin, tamoxifen, flutamide, leuprolide, goserelin, aminogluthimide, anastrozole, amsacrine, asparaginase, mitoxantrone, mitotane, amifostine or a combination thereof.

[0024] In some aspects, the tumor homing agent comprises the structure of any one of Formulas (IX), (X), (XI), (XII), (XIII), (XIV), (XV), or (XVI), wherein A4is the polypeptide:

[0025] In some aspects, the detectable agent comprises a dye, a fluorophore, a fluorescent biotin compound, a luminescent compound, a chemiluminescent compound, a radioisotope, nanoparticle, a paramagnetic metal ion, a tumor homing agent, a nerve homing agent, and a combination thereof. In some aspects, the detectable agent has a peak absorbance of 200 nm to 900 nm. In some aspects, the detectable agent has a peak emission of 300 nm to 1000 nm. In some aspects, the detectable agent has a peak emission of 400 nm to 600 nm, 450 nm to 600 nm, 500 nm to 600 nm, 550 nm to 600 nm, 400 nm to 550 nm, 400 nm to 500 nm, 400 nm to 450 nm, or 450 nm to 550 nm. In some aspects, the detectable agent has a peak emission between the 600 to 900 nm or 600 to 775 nm wavelength.

[0026] In some aspects, the dye comprises DyLight-680, DyLight-750, VivoTag-750, DyLight-800, IRDye-800, VivoTag-680, Cy5, Cy5.5, Cy7, Cy7.5, an indocyanine green (ICG), near infrared dyes, acridine orange or yellow, 7-actinomycin D, 8-anilinonaphthalene-l -sulfonic acid, ATTO dye and any derivative thereof, auramine-rhodamine stain and any derivative thereof, benzanthrone, bimane, 9-10-bis(phenylethynyl)anthracene, 5,12 — bis(phenylethynyl)naththacene, bisbenzimide, brainbow, calcein, carboxyfluorescein and any derivative thereof, 1 -chi oro-9, 10-bis(phenylethynyl)anthracene and any derivative thereof,DAPI, DiOC6, DyLight Fluors and any derivative thereof, epicocconone, ethidium bromide, FlAsH-EDT2, Fluo dye and any derivative thereof, FluoProbe and any derivative thereof, Fluorescein and any derivative thereof, Fura and any derivative thereof, GelGreen and any derivative thereof, GelRed and any derivative thereof, fluorescent proteins and any derivative thereof, m isoform proteins and any derivative thereof, heptamethine dye and any derivative thereof, hoeschst stain, iminocoumarin, indiarythrone, indo-1 and any derivative thereof, laurdan, lucifer yellow and any derivative thereof, luciferin and any derivative thereof, luciferase and any derivative thereof, mercocyanine and any derivative thereof, methylene blue and any derivative thereof, nile dyes and any derivative thereof, OS680, OS750, perylene, phloxine, phyco dye and any derivative thereof, propidium iodide, pyranine, rhodamine and any derivative thereof, ribogreen, RoGFP, rubrene, stilbene and any derivative thereof, sulforhodamine and any derivative thereof, SYBR and any derivative thereof, synapto-pHluorin, tetraphenyl butadiene, tetrasodium tris, Titan Yellow, topotecan, TSQ, umbelliferone, violanthrone, yellow fluroescent protein, YOYO-1 and ZW800, fluorescein and fluorescein dyes, carbocyanine, merocyanme, styryl dyes, oxonol dyes, phycoerythrin, erythrythrosinein. rhodamine dyes, coumarin and coumarin dyes, Oregon Green Dyes, Texas Red, Texas Red-X, SPECTRUM RED, SPECTRUM GREEN, cyanine dyes, ALEXA FLUOR dyes and any derivative thereof, BODIPY dyes, IRDyes, or any combination thereof; optionally wherein the m isoform proteins and any derivative thereof comprises mCherry; optionally wherein the fluorescein and fluorescein dyes comprise fluorescein isothiocyanate or FITC, naphthofluorescein, 4', 5' -dichloro-2',7' -dimethoxyfluorescein, or 6-carboxyfluorescein or FAM; optionally wherein the rhodamine dyes comprise carboxytetramethyl-rhodamine or TAMRA. carboxyrhodamine 6G, carboxy-X-rhodamine (ROX), lissamine rhodamine B, rhodamine 6G, rhodamine Green, rhodamine Red, or tetramethylrhodamine (TMR; optionally wherein the coumarin and coumarin dyes comprise methoxy coumarin, di alky I aminocoumarin, hydroxy coumarin, or aminomethylcoumarin (AMCA); optionally wherein the Oregon Green Dyes comprise Oregon Green 488, Oregon Green 500. or Oregon Green 514; optionally wherein the SPECTRUM GREEN comprises a cyanine dye comprising CY-3, Cy-5, CY-3.5, or CY-5.5; optionally wherein the ALEXA FLUOR dyes comprise ALEXA FLUOR 350, ALEXA FLUOR 488, ALEXA FLUOR 532, ALEXA FLUOR 546, ALEXA FLUOR 568, ALEXA FLUOR 594, ALEXA FLUOR 633, ALEXA FLUOR 660, or ALEXA FLUOR 680; optionally wherein the BODIPY dyes comprise BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650, or BODIPY 650 / 665; optionally wherein the IR Dyes comprise IRD40, IRD 700, or IRD 800;optionally wherein the radioisotope comprises iodine-131, iodine-125, bismuth-212, bismuth-213, lutetium-177, rhenium-186, rhenium-188, yttrium-90, astatine-211, phosphorus-32 and / or samarium- 153, or an isotope having one or more atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature, or any combination thereof.

[0027] In some aspects, the near infrared dye comprises a cyanine dyes. In some aspects, the dye is selected from indocyanine green (ICG), Cy5, Cy5.5, Cy7, Cy7.5, and IRDye800cw. In some aspects, the indocyanine green (ICG) has a peak absorbance of 750 nm to 770 nm, 755 nm to 775 nm, or 768 nm. In some aspects, the indocyanine green (ICG) has a peak emission of 790 nm to 820 nm, 805 nm to 815 nm, or 807 nm.

[0028] In some aspects, the isotope having one or more atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature comprises hydrogen, carbon, fluorine, phosphorous, copper, gallium, yttrium, technetium, indium, iodine, rhenium, thallium, bismuth, astatine, samarium, and lutetium, or any combination thereof. In some aspects, the lutetium comprises3H,2H,13C,14C,18F,32P,35S,64Cu,67Ga,90Y,99mTc,111In,125I,123I,131I,135I,186Re,187Re,201Tl,212Bi,211At,153Sm, or177Lu.

[0029] In some aspects, the tumor homing agent comprises a therapeutic agent selected from a radioisotope, nanoparticle, toxin, enzyme, sensitizing drug, radiosensitizer, photosensitizer, nucleic acid, interfering RNA, antibody, antibody fragment, aptamer, anti -angiogenic agent, anti-metabolite, mitotic inhibitor, grow th factor inhibitor, or any combination thereof.

[0030] In some aspects, the nerve homing agent is a molecule of Formula (DI):wherein:R1and R2are each independently selected from the group of straight or branched Ci- C6alkyl, -(CH2)m-SO3-, -(CH2)n1-N+(CH3)3, -CH2-O-X1, -CH2-CH2-O-[CH2-CH2-O]n2-X1, -CH2-CH2-CH2-O-X1, and -CH2-CH2-CH2-O-[CH2-CH2-CH2-O]n3- X1; or a moiety selected from the group of:Rs is hydrogen or R2 and Rs together form a fused ring, creating a core of FormulaR4 and Rs, together with the nitrogen atom to which they are bound, form a ring selected from the group of:or, when the compound is of Formula (DII), R4 and R5 may be independently selected from Ci-Cs alkyl, with the proviso that, when R4 is ethyl, Rs is not ethyl; Re is hydrogen;or, when R2 and R3 together form a fused ring to create a core of Formula (Dill), Rs and Re may also, together with the nitrogen atom to which Rs is bound, form afused ring, creating a core of Formula (Dill):with the proviso that, when R4 and Rs, together with the nitrogen atom to which theyare bound, form the ring, and Rs is H, Ri and R2, together with the nitrogen atom to which they are bound, may form a pyrrolidinyl ring;Xi in each instance is independently selected from C1-C6 straight or branched alkyl, Ci-Ce straight or branched alkenyl, Ci-Ce straight or branched alkynyl, and - Si(Ci-C4alkyl)3;n is an integer selected from the group of 1 and 2;nl is an integer independently selected in each instance from the group of 1, 2, 3, and 4;n2 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9. and 10;n3 is an integer independently selected in each instance from the group of 1, 2, 3. 4, 5, 6, 7, 8, 9, and 10;n4 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; andwith the proviso that the sum of n2 + n2 is not greater than 10;with the proviso that the sum of n2 + n3 is not greater than 10;with the proviso that the sum of n2 + n4 is not greater than 10;with the proviso that the sum of n3 + n4 is not greater than 10;with the proviso that, when R4 and Rs, together with the nitrogen atom to which theyare bound, form the ring. Ri and R2 are not both methyl, Ri and R2 are not both ethyl, Ri and R2 are not both n-propyl, Ri and R2 are not both n-butyl, and Ri and R2 are not both n-pentyl;with the proviso that, when the compound is of Formula (DI), when Ri is methyl, R4 is not methyl; andwith the proviso that, when the compound is of Formula (DI), when Ri is ethyl. R4 is not ethyl.

[0031] In some aspects, the nerve homing agent is a molecule of Formula (DIV):wherein:R1is C1-6 alkyl.

[0032] In some aspects, the nerve homing agent homing agent is selected from:

[0033] In some aspects, the nerve homing agent homing agent is detected at a 500-725 nm excitation. In some aspects, the nerve homing agent homing agent is detected at a 600-800 nm emission.

[0034] In some aspects, the nerve homing agent homing agent is:

[0035] In some aspects, the nerve homing agent homing agent is detected at a 655 nm peak excitation. In some aspects, the nerve homing agent homing agent is detected at a 678 nm peak emission.

[0036] In some aspects, the nerve homing agent is a molecule of Formula (El):Xi in each instance is independently selected from the group of Ci-Ce straight or branched alkyl, C2-C6straight or branched alkenyl, C1-C6straight or branched alkynyl, and -Si(C1-C4alkyl)3;nl is an integer independently selected in each instance from the group of 1, 2, 3, and 4;n2 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;n3 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9. and 10;n4 is an integer independently selected in each instance from the group of 1, 2, 3. 4, 5, 6, 7, 8, 9, and 10; andn5 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9. and 10;with the proviso that the sum of n2 + n4 is not greater than 10;with the proviso that the sum of n3 + n5 is not greater than 10;with the proviso that the sum of n2 + n5 is not greater than 10; andwith the proviso that the sum of n3 + n4 is not greater than 10.

[0037] In some aspects, the nerve homing agent homing agent is selected from:

[0038] In some aspects, the nerve homing agent homing agent is detected at a 450 to 700 nm excitation. In some aspects, the nerve homing agent homing agent is detected at a 600 to 800 nm emission. In some aspects, the nerve homing agent homing agent is detected at a 625 to 640 nm excitation. In some aspects, the nerve homing agent homing agent is detected at a 655 to 661 nm emission.

[0039] In some aspects, the nerve homing agent is a molecule of Formula (FI):wherein:one of R1and R2is hydrogen;the other of R1and R2is selected from the group of:R3and R4are independently selected from the group of H, -CN. -O-Ci-Cs alkyl, — NH2, NH(CI-C6alkyl), — N(CI-C6alkyl)2, — CH=O, and — SO2(Ci-C6alkyl); with the proviso that not more than one of R3 and R4 is hydrogen.

[0040] In some aspects, the nerve homing agent homing agent is selected from:

[0041] In some aspects, the nerve homing agent is a molecule of Formula (CI):wherein:X is selected from the group of a single bond and a double bond;Y is selected from the group of a single bond and a double bond;with the proviso that at least one of X and Y is a single bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together form a heterocyclic ring selected from the group of azetidinyl, pyrrolidinyl, and piperidinyl, wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle of the formula:n3 in each instance is an integer independently selected from the group of 0, 1, 2. and 3; n4 in each instance is an integer independently selected from the group of 0, 1, 2. and 3; with the proviso that the sum of n3 and n4 is not less than 2 and not greater than 4; n5 in each instance is an integer independently selected from the group of 0, 1, 2, and 3; n6 in each instance is an integer independently selected from the group of 0, 1, 2, and 3; with the proviso that the sum of n5 and n6 is not less than 2 and not greater than 4; R3 is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R4 is Ci-Ce alkyl when R3 is Ci-Ce alkyl; and R4 is -(CH2)ni-O-(CH2)n2-CH3 when R3 is -(CH2)nl-O-(CH2)n2-CH3;with the proviso that the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH3 moiety may be the same or different from the integers of nl and n2 in the R3 -(CH2)ni-O-(CH2)n2-CH3moiety;Rs is H; or Rs is an oxygen atom that joins with R4 to form a 2,3,4,5a,6a,lla-hexahydro-[l,4]oxazino[2,3-b]phenoxazine compound of Formula (CII):Re is a ring heteroatom selected from the group of O and S;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0042] In some aspects, the nerve homing agent homing agent is selected from:pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0043] In some aspects, the nerve homing agent is a molecule of Formula (GI):R1and R2are each independently selected from hydrogen and C1-C4 alkyl; R3aand R3bare independently selected from hydrogen and C1-C4 alkyl, with the proviso that at least one of R3aand R3bis not hydrogen; or R3aand R3btogether with the nitrogen atom to which they are bound form a 5- or 6-membered nitrogen-containing ring;or R3bis selected from hydrogen and C1-C4 alkyl, and R3atogether with the nitrogen to which it is bound and R4forms a fused 5-membered or 6-membered ring, the 5-membered or 6-membered ring having one nitrogen heteroatom, two nitrogen heteroatoms, or one nitrogen heteroatom and one oxygen heteroatom, wherein each nitrogen heteroatom is optionally substituted by a C1-C4 alkyl substituent; or R3a, the nitrogen to which it is bound, and R4form a fused 6-membered, nitrogencontaining ring and R3b, the nitrogen to which it is bound, and R2together form a fused 6-membered, nitrogen-containing ring;R4and R7are each independently selected from hydrogen, halogen, or C1-C4 alkyl; R3and R6are each independently selected from hydrogen or C1-C4 alkyl;R8aand R8bare independently selected from hydrogen and C1-C4 alkyl, with the proviso that at least one of R8aand R8bis not hydrogen; or R8aand R8btogether with the nitrogen to which they are bound form a 5- or 6-membered nitrogencontaining ring optionally substituted by 1, 2, 3, or 4 C1-C4 alkyl substituents; or R8bis selected from hydrogen and C1-C4 alkyl, and R8atogether with the nitrogen to which it is bound and R7forms a fused 6-membered ring having one nitrogen heteroatom, two nitrogen heteroatoms, or one nitrogen heteroatom and one oxygen heteroatom, wherein each nitrogen heteroatom is optionally substituted by 1, 2, 3, or 4 C1-C4 alkyl substituents;or R8a, the nitrogen to which it is bound, and R7form a fused 6-membered, nitrogencontaining ring and R8b, the nitrogen to which it is bound, and R1together form a fused 6-membered, nitrogen-containing ring;with the proviso that the compounds of Formula I do not include N3, N3, N7, N7- tetraethyl-10H-phenoxazine-3.7-diamine; N3, N7-diethyl-10H-phenoxazine-3,7- diamine; N3, N3, N7, N7-tetramethyl-10H-phenoxazine-3,7-diamine; and N7, N7- diethyl-N3, N3,2-trimethyl-10H-phenoxazine-3,7-diamine.

[0044] In some aspects, the nerve homing agent homing agent is selected from:

[0045] In some aspects, the nerve homing agent is a molecule of Formula (HI):wherein:R is a straight or branched alkyl chain of from 2 to 12 carbon atoms;R1is selected from the group of methyl, ethyl, n-propyl, isopropyl, — (CH2)ni — SO3, — (CH2)m— N+(CH3)3, — CH2— CH2— O— XI, — CH2— CH2— O—[CH2— CH2— O]n2— Xi, — CH2— CH2— CH2— O— Xi, and —CH2—CH2— CH2— O— [CH2— CH2— CH2— O]n3— Xi,Xi in each instance is independently selected from C1-C6 straight or branched alkyl.C2-C6 straight or branched alkenyl, Ci-Ce straight or branched alkynyl, and — Si(Ci-C4alkyl)3;nl is an integer independently selected in each instance from the group of 1, 2, 3, and 4;n2 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9. and 10;n3 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9. and 10;n4 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5. 6, 7, 8, 9, and 10; andn5 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5. 6, 7, 8. 9, and 10;with the proviso that the sum of n2+n4 is not greater than 10;with the proviso that the sum of n3+n5 is not greater than 10;with the proviso that the sum of n2+n5 is not greater than 10; andwith the proviso that the sum of n3+n4 is not greater than 10.

[0046] In some aspects, the nerve homing agent homing agent is selected from:

[0047] In some aspects, the nerve homing agent is a molecule of Formula (JI):wherein:R1is selected from hydrogen and C1-C3 alkyl;R2is selected from hydrogen and C1-C3 alkyl;R3is selected from the group of hydrogen, halogen, and C1-C3 alkyl:or, when R3is bound to the 9-position carbon of the 10H-benzo[c]phenoxazine core, R2and R3, together with the carbon atoms to which they are bonded, form a fivemembered or six-membered fused ring, the five-membered or six-membered fused ring optionally containing a ring oxygen heteroatom, and the fivemembered or six-membered fused ring being substituted by 0. 1, 2, or 3 Ci- C3 alkyl substituents; andR4and R5are each independently selected from hydrogen and C1-C3 alkyl.

[0048] In some aspects, the nerve homing agent homing agent is selected from:

[0049] In some aspects, the nerve homing agent homing agent is detected at a 450-800 nm excitation. In some aspects, the nene homing agent homing agent is detected at a 550-850 nm emission. In some aspects, the nerve homing agent homing agent is detected at a 540-760 nm excitation. In some aspects, the nerve homing agent homing agent is detected at a 605-785 nm emission.

[0050] In some aspects, the tumor homing agent and the nerve homing agent each detect a physiologic structure or cell between at least 10 pm and 25 pm in diameter. In some aspects, the nerve homing agent detects a physiologic structure or cell between at least 10 pm and 25 pm in diameter.

[0051] In some aspects, the method further comprises causing remission in, reducing, ameliorating, or ablating the cancer or vascular lesion, include remission, increasing quality of life, reducing the extent of resection, extending progression free survival, or number of surgeries required, reducing pain, or exhibiting other clinical or quality of life benefit to a patient.

[0052] Some embodiments are directed towards a method of detecting a tissue or cell and a nerve tissue or cell in a subject in need thereof, the method comprising administering to a subject a tumor homing agent and a nerve homing agent.

[0053] In some embodiments, the detecting comprises fluorescence imaging.

[0054] In some embodiments, the tumor homing agent is fluorescent, the nerve homing agent is fluorescent, or both.

[0055] In some embodiments, the method comprises performing a fluorescence guided surgery (FGS) procedure on the subject in need thereof.

[0056] In some embodiments, the minimum spectral gap in emission wavelength of the nerve homing agent relative to the tumor homing agent is at least about 0 nm to at least about 800 nm.

[0057] In some embodiments, the minimum spectral gap in emission wavelength of the nerve homing agent relative to the tumor homing agent is at least about 0 nm, at least about 10 nm, at least about 20 nm, at least about 30 nm, at least about 40 nm, at least about 50 nm, at least about 60 nm, at least about 70 nm, at least about 80 nm, at least about 90 nm. at least about 100 nm, at least about 110 nm, at least about 120 nm, at least about 130 nm, at least about 140 nm, at least about 150 nm, at least about 160 nm, at least about 170 nm, at least about 180 nm, at least about 190 nm, at least about 200 nm.

[0058] In some embodiments, the emission wavelength spectrum of the nerve homing agent overlaps the excitation wavelength spectrum of the tumor homing agent by at least about 0%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%. at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%. at least about 18%. at least about 19%, at least about 20%, at least about 21%, at least about 22%, at least about 23%, at least about 24%, at least about 25%, at least about 26%, at least about 27%, at least about 28%, at least about 29%, at least about 30%, at least about 31%, at least about 32%, at least about 33%, at least about 34%, atleast about 35%, at least about 36%, at least about 37%, at least about 38%, at least about 39%, at least about 40%.

[0059] In some embodiments, a spectral gap in peak excitation wavelength of the nerve homing agent and tumor homing agent is at least about 400 nm to at least about 800 nm.

[0060] In some embodiments, the minimum spectral gap in emission wavelength of the nerve homing agent relative to the tumor homing agent is at least about 0 nm, at least about 10 nm, at least about 20 nm, at least about 30 nm, at least about 40 nm, at least about 50 nm, at least about 60 nm, at least about 70 nm, at least about 80 nm, at least about 90 nm, at least about 100 nm, at least about 110 nm, at least about 120 nm. at least about 130 nm, at least about 140 nm, at least about 150 nm, at least about 160 nm, at least about 170 nm, at least about 180 nm, at least about 190 nm, at least about 200 nm.

[0061] In some embodiments, the excitation spectrum of the nerve homing agent does not overlap the spectrum emission wavelength of the tumor homing agent.

[0062] In some embodiments, the excitation spectrum of the nerve homing agent completely overlaps the spectrum emission wavelength of the tumor homing agent.

[0063] In some embodiments, the emission spectrum of the nerve homing agent does not overlap the spectrum excitation wavelength of the tumor homing agent.

[0064] In some embodiments, the emission spectrum of the nerve homing agent completely overlaps the spectrum excitation wavelength of the tumor homing agent.

[0065] In some embodiments, the fluorescence guided surgery (FGS) is performed with the assistance of an imaging system.

[0066] In some embodiments, the imaging system is selected from the group consisting of: surgical microscope, confocal microscope, fluorescence scope, exoscope, endoscope, ophthalmoscope, retinal camera system, optical coherence tomography (OCT) system, surgical robot, and Quest Spectrum imaging system, Fluobeam devices, Lab Flare imaging sy stem, or Solaris imaging system, or other imaging system equipped with two-color imaging capabilities.

[0067] In some embodiments, the nerve homing agent is retained by. or accumulates in and / or binds to a nerve tissue or nerve cell.

[0068] Some embodiments further comprise detecting the presence or absence of the nerve homing agent in a tissue or cell, wherein the presence of the nerve homing agent in the tissue or cell indicates the presence of nerve tissue or a nerve cell.

[0069] In some embodiments, the tumor homing agent is retained by, or accumulates in and / or binds to cancerous tissue or a cancer cell, or a vascular lesion.

[0070] Some embodiments further comprise detecting the presence or absence of the tumor homing agent in a tissue or cell, wherein the presence of the tumor homing agent in the tissue or cell indicates the presence of a cancerous tissue or a cancer cell, or a vascular lesion.

[0071] Some embodiments further comprise treating the cancer or vascular lesion in the subject in need thereof.

[0072] In some embodiments, the treating further comprises surgically removing a cancerous tumor, a cancerous tissue, or a cancerous cell, or vascular lesion, from the subject in need thereof by performing a surgery.

[0073] In some embodiments, the surgery is fluorescence guided surgery (FGS).

[0074] In some embodiments, surgically removing the cancerous tumor, the cancerous tissue, or the cancerous cell, or vascular lesion, from the subject in need thereof spares the nerve tissue or nerve cell, avoids damaging the nerve tissue or nerve cell, or does not damage a nerve tissue or nerve cell in the subject in need thereof.

[0075] In some embodiments, the surgically removed the cancerous tumor, the cancerous tissue, or the cancerous cell, or vascular lesion, from the subject in need thereof shows sparing the nerve tissue or nerve cell, avoiding damaging the nerve tissue or nerve cell, or not damaging a nerve tissue or nerve cell ex vivo in pathology samples obtained from the subject in need thereof or in situ in the patient in need thereof.

[0076] In some embodiments, the cancer is glioma, astrocytoma, medulloblastoma, choroids plexus carcinoma, ependymoma, neuroblastoma, vestibular schwannoma, carcinoma, meningioma, head and neck cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, breast cancer, intestinal cancer, pancreatic cancer, liver cancer, kidney cancer, sarcoma, osteosarcoma, rhabdomyosarcoma, Ewing’s sarcoma, gastrointestinal stromal tumors, melanoma, ovarian cancer, cervical cancer, lymphoma, thyroid cancer, anal cancer, colorectal cancer, endometrial cancer, laryngeal cancer, multiple myeloma, prostate cancer, retinoblastoma, gastric cancer, testicular cancer. Wilm’s tumor, or any combination thereof, or where the vascular lesion is a cavernous angioma, cavernous hemangioma, or cerebral cavernous malformation (CCM), or any combination thereof.

[0077] In some embodiments, the cancer is head and neck cancer, a brain cancer, a brain tumor, breast cancer, melanoma, sarcoma, basal cell carcinoma, squamous cell carcinoma, lung cancer, colorectal cancer, prostate cancer, bladder cancer, or any combination thereof.

[0078] In some embodiments, the head and neck cancer is head and neck squamous cell carcinoma.

[0079] In some embodiments, the breast cancer is invasive ductal carcinoma, ductal carcinoma in situ breast cancer, invasive lobular carcinoma, lobular carcinoma in situ breast cancer, triple negative breast cancer, or any combination thereof.

[0080] In some embodiments, the tumor homing agent is formulated for intravenous administration, intravenous bolus administration, intravenous infusion administration, intramuscular administration, subcutaneous administration, intraperitoneal administration, oral administration, sublingual administration, topical administration, transdermal administration using electroporation, intralesional administration, or a combination thereof.

[0081] In some embodiments, the nerve homing agent is formulated for intravenous administration, intravenous bolus administration, intravenous infusion administration, intramuscular administration, subcutaneous administration, intraperitoneal administration, oral administration, sublingual administration, topical administration, transdermal administration using electroporation, intralesional administration, or a combination thereof.

[0082] In some embodiments, the tumor homing agent and the nerve homing agent are coadministered.

[0083] In some embodiments, the tumor homing agent and the nerve homing agent are sequentially administered.

[0084] In some embodiments, the tumor homing agent is administered 1 to 24 hours before, to 1-3 days before a surgery and the nerve homing agent is administered 10-120 minutes before, to 2 to 48 hours before the same surgery.

[0085] In some embodiments, the tumor homing agent is administered prior to the nerve homing agent, or wherein the nerve homing agent is administered prior to the tumor homing agent, or wherein the tumor homing agent and the nerve homing agent are administered at the same time.

[0086] In some embodiments, the tumor homing agent comprises a polypeptide comprising: (a) a sequence of any one of SEQ ID NO: 482 - SEQ ID NO: 485, or a fragment thereof, (b) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512, or a fragment thereof, or (c) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 9, or a fragment thereof.

[0087] In some embodiments, the polypeptide comprises SEQ ID NO: 9, or a fragment thereof.

[0088] In some embodiments, the fragment of the polypeptide has a length of at least 25 residues.

[0089] In some embodiments, each amino acid of the polypeptide is independently selected as an L- or D-enantiomer.

[0090] In some embodiments, the polypeptide contains no lysine residues.

[0091] In some embodiments, the polypeptide contains a single lysine residue.

[0092] In some embodiments, the single lysine residue is located at a position corresponding to K-27 of native chlorotoxin, K-23 of native chlorotoxin, or K-15 of native chlorotoxin.

[0093] In some embodiments, one, two, or three methionine residues of the polypeptide are replaced with other amino acids.

[0094] In some embodiments, the N-terminus of the polypeptide is blocked by acetylation or cyclization.

[0095] In some embodiments, the polypeptide comprises at least 4. at least 5, or at least 6 disulfide bonds.

[0096] In some embodiments, the polypeptide has a length of no less than 21 amino acid residues, no less than 22 amino acid residues, no less than 23 amino acid residues, no less than 24 amino acid residues, no less than 25 amino acid residues, no less than 26 amino acid residues, no less than 27 amino acid residues, no less than 28 amino acid residues, no less than 29 amino acid residues, no less than 30 amino acid residues, no less than 31 amino acid residues, no less than 32 amino acid residues, no less than 33 amino acid residues, no less than 34 amino acid residues, no less than 5 amino acid residues, no less than 36 amino acid residues, no less than 37 amino acid residues, no less than 38 amino acid residues, no less than 39 amino acid residues, or no less than 40 amino acid residues.

[0097] In some embodiments, the polypeptide comprises an isoelectric point at least 8.0, at least 8 5, or at least 9.0,

[0098] In some embodiments, the polypeptide is conjugated to an agent.

[0099] In some embodiments, the agent is a detectable agent or a first therapeutic agent.

[0100] In some embodiments, the polypeptide is conjugated to the agent via a cleavable linker or a stable linker

[0101] In some embodiments, the polypeptide comprises: (a) a single lysine residue and the agent is conjugated to the polypeptide at the single lysine residue, or (b) no lysine residues and the agent is conjugated to the polypeptide at the N-terminus of the polypeptide.

[0102] In some embodiments, the polypeptide and agent comprises the structure of Formula (I), or a pharmaceutically acceptable salt thereof:R1, R2, R3, R4, R5, R6, R7, R8, R15, and R16are each independently selected from hydrogen. Ci-Ce alkyl. Ci-Cs alkylene-COOH, sulfonate, Ci-Ce alkylene-sulfonate, -COOH, -SO2-NH2, or Ci-Ce alkoxy;R9is hydrogen, sulfonate, amine, or COOH;L1is C3-C6 alkylene;L² is C1-C10 alkylene;L3is a bond, -O-, -NR10-, -NR10-Ci-C6alkylene- -O-NR10-, - R10-CI-C6alkylene- (O-Ci-Ce alkylene)n-, -NR10-L4-, - R10-CI-C6 alkylene-NR11- (C (= O) -Ci-Ce alkylene-O-) -, or -NR10-CI-C6 alky lene— NR10-Ci-Ce. alkylene— NR!0-CI-C6alkylene-;L4is a bond, -heterocyclyl ---. or -heterocyclyl-Ci-Cs alkylene-;R10is hydrogen or Ci-Ce alkyl;R11is hydrogen or Ci-Cs alkyl;R12and R13are independently selected from hydrogen, Ci-Ce alkyl, or R12and R13are joined together along with the other atoms to which they’ are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R14is hydrogen or C1-C.5 alkylene, -(L’j-aryl, -(L )-aryl-R21, -(L5)-heteroaiyl, -(L5)-heteroaryl-R2!, -NR17R18, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;L5is a bond, C1-C10 alkylene, O, -NR10-;R17and R18are each independently hydrogen or aryl;R19and R20are independently' selected from hydrogen, Ci-Ce alkyl. R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5 -membered or 6-membered carbocyclic or heterocyclic ring;R21is hydrogen, sulfonate, or -COOH;n is 0, 1, 2, or 3;m is 0, 1, 2, or 3;p is 0, 1, 2, or 3;q is 0. 1, 2, or 3; andA4is the polypeptide.

[0103] In some embodiments, R9is sulfonate.

[0104] In some embodiments, A4has at least 90% sequence identity with: (a) a sequence of any one of SEQ ID NO: 482 - SEQ ID NO: 485, or a fragment thereof, (b) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512, or a fragment thereof, or (c) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 9, or a fragment thereof.

[0105] In some embodiments, A4is SEQ ID NO: 9 or a fragment thereof.

[0106] In some embodiments. A4comprises at least 25 amino acid residues.

[0107] In some embodiments, R3, R4, R5, R6of the Formula (I), are each independently methyl; R1, R2, R7, R8, R15, and R16are each independently hydrogen; R10is hydrogen; R12, R13, R14, R19, and R20are each independently hydrogen; L1is butylene; and L2is pentylene.

[0108] In some embodiments, L3is attached to A4at a lysine amino acid residue of the polypeptide.

[0109] In some embodiments, Formula (I) is conjugated to polyethylene glycol (PEG), hydroxy ethyl starch, polyvinyl alcohol, a water soluble polymer, a zwitterionic water soluble polymer, a water soluble poly(amino acid), an albumin, or a fatty acid.

[0110] In some embodiments. Formula (II) further comprises a second therapeutic agent attached to A4, wherein the second therapeutic agent is selected from a radioisotope, toxin, cytotoxic agent, enzy me, sensitizing drug, nucleic acid, interfering RNA, antibody, anti-angiogenic agent, cisplatin, anti-metabolite, mitotic inhibitor, growth factor inhibitor, paclitaxel, temozolomide, topotecan, fluorouracil, vincristine, vinblastine, procarbazine, dacarbazine, altretamine, methotrexate, mercaptopurine, thioguanine, fludarabine phosphate, cladribine, pentostatin, cytarabine, azacitidine, etoposide, teniposide, irinotecan, docetaxel, doxorubicin, daunorubicin, dactinomycin, idarubicin, plicamycin, mitomycin, bleomycin, tamoxifen,flutamide, leuprolide, goserelin, aminogluthimide, anastrozole, amsacrine, asparaginase, mitoxantrone, mitotane, amifostine or a combination thereof.

[0111] In some embodiments, the polypeptide and agent comprises the structure of any one of Formulas (IX), (X), (XI), (XII), (XIII), (XIV). (XV), or (XVI), wherein A4is the polypeptide:

[0112] In some embodiments, the detectable agent comprises a dye, a fluorophore, a fluorescent biotin compound, a luminescent compound, a chemiluminescent compound, a radioisotope, nanoparticle, a paramagnetic metal ion, a tumor homing agent, a nerve homing agent, and a combination thereof.

[0113] In some embodiments, the detectable agent (e.g., a dye, a fluorophore, a fluorescent biotin compound, a luminescent compound, a chemiluminescent compound) has a peak absorbance of about 200 nm to about 900 nm.

[0114] In some embodiments, the detectable agent (e.g., a dye, a fluorophore, a fluorescent biotin compound, a luminescent compound, a chemiluminescent compound) has a peak emission of about 300 nm to about 1000 nm.

[0115] In some embodiments, the detectable agent has a peak emission of about 400 nm to about 600 nm, about 450 nm to about 600 nm, about 500 nm to about 600 nm, about 550 nm to about 600 nm, about 400 nm to about 550 nm, about 400 nm to about 500 nm, about 400 nm to about 450 nm, or about 450 nm to about 550 nm.

[0116] In some embodiments, the detectable agent (e.g., a dye, a fluorophore, a fluorescent biotin compound, a luminescent compound, a chemiluminescent compound) has a peak emission between the 600-900 nm or 600-775 nm wavelength.

[0117] In some embodiments, the tumor homing agent has a peak emission between 600-900 nm inclusive.

[0118] In some embodiments, the nerve homing agent has a peak emission between 600-775 nm inclusive.

[0119] In some embodiments, the dye comprises DyLight-680, DyLight-750, VivoTag-750, DyLight-800, IRDye-800, VivoTag-680, Cy5, Cy5.5, Cy7, Cy7.5, an indocyanine green (ICG), near infrared dyes, acridine orange or yellow, 7-actinomycin D, 8-anilinonaphthalene-l -sulfonic acid, ATTO dye and any derivative thereof, auramine-rhodamine stain and any derivative thereof, benzanthrone, bimane, 9-10-bis(phenylethynyl)anthracene, 5,12 -bis(phenylethynyl)naththacene, bisbenzimide, brainbow, calcein, carboxyfluorescein and anyderivative thereof, l-chloro-9,10-bis(phenylethynyl)anthracene and any derivative thereof, DAPI, DiOC6, DyLight Fluors and any derivative thereof, epicocconone, ethidium bromide, FlAsH-EDT2, Fluo dye and any derivative thereof, FluoProbe and any derivative thereof, Fluorescein and any derivative thereof, Fura and any derivative thereof, GelGreen and any derivative thereof, GelRed and any derivative thereof, fluorescent proteins and any derivative thereof, m isoform proteins and any derivative thereof, heptamethine dye and any derivative thereof, hoeschst stain, iminocoumarin, indiarythrone, indo-1 and any derivative thereof, laurdan, lucifer yellow and any derivative thereof, luciferin and any derivative thereof, luciferase and any derivative thereof, mercocyanine and any derivative thereof, methylene blue and any derivative thereof, nile dyes and any derivative thereof, OS680, OS750, perylene, phloxine, phyco dye and any derivative thereof, propidium iodide, pyranine, rhodamine and any derivative thereof, ribogreen, RoGFP, rubrene, stilbene and any derivative thereof, sulforhodamine and any derivative thereof, SYBR and any derivative thereof, synapto-pHluorin, tetraphenyl butadiene, tetrasodium tris, Titan Yellow, topotecan, TSQ, umbelliferone, violanthrone, yellow fluroescent protein, YOYO-1 and ZW800, fluorescein and fluorescein dyes, carbocyanine, merocyanine, styryl dyes, oxonol dyes, phycoery thrin, erythrosinein. rhodamine dyes, coumarin and coumarin dyes. Oregon Green Dyes. Texas Red, Texas Red-X, SPECTRUM RED. SPECTRUM GREEN, cyanine dyes, ALEXA FLUOR dyes and any derivative thereof, BODIPY dyes, IRDyes, or any combination thereof.

[0120] In some embodiments, the near infrared dye comprises a cyanine dyes.

[0121] In some embodiments, the dye is selected from indocyanine green (ICG), Cy5, Cy5.5, Cy7, Cy7.5, and IRDye800cw.

[0122] In some embodiments, the indocyanine green (ICG) has a peak absorbance of 750 nm-770 nm, 755 nm-775 nm, or 768nm.

[0123] In some embodiments, the indocyanine green (ICG) has apeak emission of 790 nm-820 nm, 805 nm-815 nm. or 807nm.

[0124] In some embodiments, the m isoform proteins and any’ derivative thereof comprises mCherry.

[0125] In some embodiments, the fluorescein and fluorescein dyes comprise fluorescein isothiocyanate or FITC, naphthofluorescein, 4', 5' -dichloro-2',7' -dimethoxyfluorescein, or 6-carboxyfluorescein or FAM.

[0126] In some embodiments, the rhodamine dyes comprise carboxytetramethyl-rhodamine or TAMRA, carboxyrhodamine 6G, carboxy-X-rhodamine (ROX), lissamine rhodamine B, rhodamine 6G, rhodamine Green, rhodamine Red, or tetramethylrhodamine (TMR).

[0127] In some embodiments, the coumarin and coumarin dyes comprise methoxy coumarin, dialkylaminocoumarin, hydroxy coumarin, or aminomethylcoumarin (AMCA).

[0128] In some embodiments, the Oregon Green Dyes comprise Oregon Green 488, Oregon Green 500, or Oregon Green 514.

[0129] In some embodiments, the SPECTRUM GREEN comprises a cyanine dye comprising CY-3, Cy-5, CY-3.5, or CY-5.5.

[0130] In some embodiments, the ALEXA FLUOR dyes comprise ALEXA FLUOR 350, ALEXA FLUOR 488, ALEXA FLUOR 532, ALEXA FLUOR 546, ALEXA FLUOR 568, ALEXA FLUOR 594, ALEXA FLUOR 633, ALEXA FLUOR 660, or ALEXA FLUOR 680.

[0131] In some embodiments, the BODIPY dyes comprise BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650, or BODIPY 650 / 665.

[0132] In some embodiments, the IR Dyes comprise IRD40, IRD 700, or IRD 800.

[0133] In some embodiments, the radioisotope comprises iodine-131, iodine-125, bismuth-212, bismuth-213, lutetium-177, rhenium-186, rhenium-188, yttrium-90, astatine-211, phosphorus-32 and / or samarium-153, or an isotope having one or more atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature.

[0134] In some embodiments, the isotope having one or more atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature comprises hydrogen, carbon, fluorine, phosphorous, copper, gallium, yttrium, technetium, indium, iodine, rhenium, thallium, bismuth, astatine, samarium, and lutetium, or any combination thereof.

[0135] In some embodiments, the lutetium comprises ³H, ¹³C, ¹⁴C, ¹⁸F, ³²P, ³⁵S, ⁶⁴Cu, ⁶⁷Ga, ⁹⁰Y, ⁹⁹ᴹTc, ¹¹¹In, ¹²⁵I, ¹²³I, ¹³¹I, ¹³⁵I, ¹⁸⁶Re, ¹⁸⁷Re, ²⁰¹Tl, ²¹²Bi, ²¹¹At, ¹⁵³Sm, or ¹⁷⁷Lu.

[0136] In some embodiments, the first therapeutic agent comprises a radioisotope, nanoparticle, toxin, enzyme, sensitizing drug, radiosensitizer, photosensitizer, nucleic acid, interfering RNA, antibody, antibody fragment, aptamer, anti-angiogenic agent, anti-metabolite, mitotic inhibitor, growth factor inhibitor, or a combination thereof.

[0137] In some embodiments, the radioisotope comprises iodine-131, iodine-125, bismuth-212, bismuth-213, lutetium-177, rhenium-186, rhenium-188, yttrium-90, astatine-211, phosphorus-32 and / or samarium-153, or an isotope having one or more atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature, or any combination thereof.

[0138] In some embodiments, the isotope having one or more atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature comprises hydrogen, carbon, fluorine, phosphorous, copper, gallium, yttrium, technetium, indium, iodine, rhenium, thallium, bismuth, astatine, samarium, and lutetium, or any combination thereof.

[0139] In some embodiments, the lutetium comprises ³H, ¹³C, ¹⁴C, ¹⁸F, ³²P, ³⁵S, ⁶⁴Cu, ⁶⁷Ga, ⁹⁰Y, ⁹⁹ᴹTc, ¹¹¹In, ¹²⁵I, ¹²³I, ¹³¹I, ¹³⁵I, ¹⁸⁶Re, ¹⁸⁷Re, ²⁰¹Tl, ²¹²Bi, ²¹¹At, ¹⁵³Sm, or ¹⁷⁷Lu.

[0140] In some embodiments, the tumor homing agent is detected at a 715 - 800 nm excitation.

[0141] In some embodiments, the tumor homing agent is detected at a 815 - 875 nm emission.

[0142] In some embodiments, the tumor homing agent detects a physiologic structure or cell between at least 10 pm and 25 pm in diameter.

[0143] In some embodiments, the nerve homing agent is a molecule of Formula (CI):wherein:X is selected from the group of a single bond and a double bond;Y is selected from the group of a single bond and a double bond;with the proviso that at least one of X and Y is a single bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety’ of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together form a heterocyclic ring selected from the group of azetidinyl, pyrrolidinyl, and piperidinyl, wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle of the formula:the wavy linein the Ri and R2 heterocyclic spirocycle formulas represents the optional single bond or double bond through which Ri is bound to the adjacent 3- position carbon atom;n3 in each instance is an integer independently selected from the group of 0, 1, 2, and 3;n4 in each instance is an integer independently selected from the group of 0, 1, 2, and 3;with the proviso that the sum of n3 and n4 is not less than 2 and not greater than 4; n5 in each instance is an integer independently selected from the group of 0, 1, 2, and 3;n6 in each instance is an integer independently selected from the group of 0, 1, 2, and 3;with the proviso that the sum of n5 and n6 is not less than 2 and not greater than 4; R3 is selected from the group of C1-C6 alkyl and a moiety of the formula -(CH2)ni-O- (CH2)n2-CH3;R4 is Ci-Ce alkyl when R3 is Ci-Ce alkyl; and R4 is -(CH2)ni-O-(CH2)n2-CH3 when R3 is -(CH2)ni-O-(CH2)n2-CH3;with the proviso that the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH3 moiety may be the same or different from the integers of nl and n2 in the R3 - (CH2)ni-O-(CH2)n2-CH3 moi ety:Rs is H; or Rs is an oxygen atom that joins with R4 to form a 2, 3, 4, 5a, 6a, 1 la- hexahydro-[l,4]oxazino[2,3-b]phenoxazine compound of Formula (CII):Rs is a ring heteroatom selected from the group of O and S;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0144] It is understood that Formula (CI) includes each of its resonance structures as noted below and may be represented by each. For each embodiment herein concerning a compound of Formula (I) described herein, three additional separate embodiments exist wherein all variables(Ri, R2, R3, Rs, Rs, ni, etc.) are as defined for the embodiment in question and the three additional embodiments are directed to, respectively, a compound of Formula (Cla), acompound of Formula (CIb), and a compound of Formula (CIc), or a pharmaceuticallyacceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates,or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptableprodrug thereof.

[0145] A specific example of this resonance can also be seen in the set of compounds below.include those selected from the following group:wherein the wavy line through parallel straight and dashed lines ( ) and a wavy line through astraight lineeach represent the optional single and double bonds through which the heterocyclic spirocycle may be bound to the adjacent carbon atom.

[0147] In view' of the resonance structures for Formula (CI) and the other formulas herein, it is further understood that the cyclic and spirocyclic groups formed by Ri and R2, along with the nitrogen atom they are bound to. and R3 and R4, along with the nitrogen atom to which they are bound, may be represented equally in structures herein with a single bond line ( - ). a partially dashed single / double bond ( - ), and / or a double lined double bond (^=), such as represented in the structures below'.

[0148] In some embodiments, the nerve homing agent is a molecule of Formula (CIII):wherein:X is selected from the group of a single bond and a double bond;Y is selected from the group of a single bond and a double bond;with the proviso that at least one of X and Y is a single bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3;n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together with the nitrogen atom to which they are bound form a heterocyclic ring selected from the group of azetidinyl, pyrrolidinyl, and piperidinyl, wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle selected from the group of:\Zthe wavy line through a straight line \ in the Ri and R2 heterocyclic spirocycle formulas represents the optional single bond or double bond through which Ri is bound to the adjacent carbon atom;R3 is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CFbjni-O- (CH2)n2-CH3;R4 is C1-C3 alkyd when Rs is C1-C3 alkyl; and R4 is -(CH2)ni-O-(CH2)n2-CH3 when R3 is -(CH2)nl-O-(CH2)n2-CH;with the proviso that the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH3 moiety may be the same or different from the integers of nl and n2 in the R3 - (CH2)ni-O-(CH2)n2-CH3 moiety; andRs is H: or R5 is an oxygen atom that joins with R4 to form a 2,3,4,5a.6a,l la- hexahydro-[l,4]oxazino[2,3-b]phenoxazine compound of Formula (CIV):or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0149] A further embodiment provides a compound of Formula (CIII), wherein X, Y, R3, R4, and Rs are as defined in the embodiment immediately above, and Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle selected from the group of:or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0150] A further embodiment provides a compound of Formula (CIII), wherein X, Y, R3, R4, and Rs are as defined in the embodiment immediately above, and Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle selected from the group of:or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0151] Another embodiment provides a compound of Formula (CV):wherein:X is selected from the group of a single bond and a double bond;Y is selected from the group of a single bond and a double bond;with the proviso that at least one of X and Y is a single bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together with the nitrogen atom to which they are bound form a heterocyclic ring selected from the group of azetidinyl and pyrrolidinyl. wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle selected from the group of:the wavy line through a straight linein the Ri and R2 heterocyclic spirocycle formulas represents the optional single bond or double bond through which Ri is bound to the adjacent carbon atom;R3 is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CFbjni-O- (CH2)n2-CH3;R4 is C1-C3 alky l when Ra is C1-C3 alky l; and R4 is -(CH2)ni-O-(CH2)n2-CH3 when R3 is -(CH2)ni-O-(CH2)n2-CH3;with the proviso that the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH3moiety may be the same or different from the integers of nl and n2 in the R3 - (CH2)ni-O-(CH2)n2-CH3 moiety; andRs is H; or R5 is an oxygen atom that joins with R4 to form a 2,3,4,5a.6a,l la- hexahydro-[l,4]oxazino[2,3-b]phenoxazine compound of Formula (CVI):or a pharmaceutically acceptable salt, co-crvstal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0152] Another embodiment provides a compound of Formula (CVII):wherein:X is selected from the group of a single bond and a double bond;Y is selected from the group of a single bond and a double bond;with the proviso that at least one of X and Y is a single bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R.2 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together with the nitrogen atom to which they are bound form a heterocyclic ring selected from the group of azetidinyl and pyrrolidinyl. wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a heterocyclic spirocycle selected from the group of:the wavy linein the Ri and R2 heterocyclic spirocycle formulas represents the optional single bond or double bond through which Ri is bound to the adjacent 3- position carbon atom;Rs is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CH2)ni-O- (CH2)n2-CH3;R4 is C1-C3 alkyl when R3 is C1-C3 alkyl; and R4 is -(CH2)ni-O-(CH2)n2-CH3 when R3 is -(CH2)nl-O-(CH2)n2-CH3;with the proviso that the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH3moiety may be the same or different from the integers of nl and n2 in the R3 -(CH2)n1-O-(CH2)n2-CH3 moiety; andR5 is H; or Rs is an oxygen atom that joins with R4 to form a 2,3,4,5a,6a,lla-hexahydro- [l,4]oxazino[2,3-b]phenoxazine compound of Formula (CVIII):or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0153] A further embodiment provides a compound of Formula (CIX):wherein:X is selected from the group of a single bond and a double bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together form a heterocyclic ring selected from the group of azetidinyl, pyrrolidinyl, and piperidinyl. wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle selected from the group of:the wavy line through a straight linein the Ri and R2 heterocyclic spirocycle formulas represents the optional single bond or double bond through which Ri is bound to the adjacent carbon atom;andR3 is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CH2)ni-0- (CH2)n2-CH3;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0154] A further embodiment provides a compound of Formula (CX):wherein:X is selected from the group of a single bond and a double bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together form a heterocyclic ring selected from the group of azetidinyl, pyrrolidinyl, and piperidinyl, wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a heterocyclic spirocycle selected from the group of:the wavy linein the Ri and R2 heterocyclic spirocycle formulas represents the optional single bond or double bond through which Ri is bound to the adjacent 3- position carbon atom; andR. is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CFbjni-O- (CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3;n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0155] A further embodiment provides a compound of Formula (CXI):wherein:X is selected from the group of a single bond and a double bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together form a heterocyclic ring selected from the group of azetidinyl, pyrrolidinyl, and piperidinyl. wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle selected from the group of:the wavy line through a straight linein the Ri and R2 heterocyclic spirocycle formulas represents the optional single bond or double bond through which Ri is bound to the adjacent carbon atom; andR3 is Ci-Ce alkyl; andR4 is Ci-Ce alkyl;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0156] A further embodiment provides a compound of Formula (CXII):wherein:X is selected from the group of a single bond and a double bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together form a heterocyclic ring selected from the group of azetidinyl, pyrrolidinyl, and piperidinyl, wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle selected from the group of:the wavy line through a straight linein the Ri and R2 heterocyclic spirocycle formulas represents the optional single bond or double bond through which Ri is bound to the adjacent carbon atom;R3 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3; andR4 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0157] A further embodiment provides a compound of Formula (CXIII):wherein:X is selected from the group of a single bond and a double bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety of the formula (CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together form a heterocyclic ring selected from the group of azetidinyl, pyrrolidinyl, and piperidinyl, wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle selected from the group of:the wavy linein the Ri and R2 heterocyclic spirocycle formulas represents the optional single bond or double bond through which Ri is bound to the adjacent 3- position carbon atom;Rs is a moiety of the formula -(CH2)ni-O-(CH2)n2-CHs; andR4 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CHs;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0158] A further embodiment provides a compound of Formula (CXIV):wherein:X is selected from the group of a single bond and a double bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CHs;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together form a heterocyclic ring selected from the group of azetidinyl, pyrrolidinyl, and piperidinyl, wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle selected from the group of:the wavy linein the Ri and R2 heterocyclic spirocycle formulas represents the optional single bond or double bond through which Ri is bound to the adjacent 3- position carbon atom;R3 is Ci-Ce alky l: andR4 is Ci-Ce alkyl:or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0159] A further embodiment provides a compound of Formula (CXIV) wherein X, Ri, and R2 are as defined above; nl in each instance is an integer independently selected from the group of 1 and 2; and n2 in each instance is an integer independently selected from the group of 0 and 1; R3 is C1-C4 alkyl; and R4 is C1-C4 alkyl; or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0160] A still further embodiment provides a compound of Formula (CXIV) wherein X, Ri, and R2 are as defined above; nl in each instance is an integer independently selected from the group of 1 and 2; and n2 in each instance is an integer independently selected from the group of 0 and 1; R3 is C1-C3 alkyl; and R4 is C1-C3 alkyl; or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0161] Another embodiment provides a compound of Formula (CXV):wherein:R3 is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CH2)ni-O- (CH2)n2-CH3;nl is an integer independently selected from the group of 1, 2, and 3; and n2 is an integer independently selected from the group of 0, 1, and 2;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0162] Another embodiment provides a compound of Formula (CXVI):wherein:R3 is selected from the group of Ci-Ce alkyl and a moiety of the formula (CH2)ni-0- (CH2)n2-CH3;nl is an integer independently selected from the group of 1, 2. and 3; and n2 is an integer independently selected from the group of 0, 1, and 2;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0163] Another embodiment provides a compound of Formula (CXVII):wherein:R3 is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CH2)ni-O- (CH2)n2-CH3;nl is an integer independently selected from the group of 1, 2, and 3;n2 is an integer independently selected from the group of 0, 1, and 2;R9 is selected from the group of methyl and ethyl; andRio is selected from the group of methyl and ethyl;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0164] Another embodiment provides a compound of Formula (CXVIII):wherein:R3 is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CH2)ni-0- (CH2)n2-CH3; andR7 is selected from the group of methoxy and ethoxy;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0165] Another embodiment provides a compound of Formula (CXIX):wherein:R3 is selected from the group of Ci-Ce alky l and a moiety of the formula -(CH2)ni-O- (CH2)n2-CH3;R7 is selected from the group of methoxy and ethoxy; andR7 is selected from the group of H, methoxy and ethoxy;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0166] Another embodiment provides a compound of Formula (CXX):wherein:Ra is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CH2)ni-O- (CH2)n2-CH3;nl is an integer independently selected from the group of 1, 2, and 3; and n2 is an integer independently selected from the group of 0, 1, and 2;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0167] Another embodiment provides a compound of Formula (CXXI):wherein:R3 is selected from the group of Ci-Ce alkyd and a moiety of the formula (CH2)ni-O- (CH2)n2-CH3;nl is an integer independently selected from the group of 1, 2. and 3; and n2 is an integer independently selected from the group of 0, 1, and 2;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0168] Further separate embodiments provide a compound, respectively, of Formula (CXV), Formula (CXVI), Formula (CXVII), Formula (CXVIII), Formula (CXVIX), Formula (CXX), and Formula (CXXI), wherein, in each separate embodiment:Rs is selected from the group of C1-C4 alkyl and a moiety of the formula -(CH2)ni-O- (CH2)n2-CH3;nl is an integer independently selected from the group of 1, 2, and 3; and n2 is an integer independently selected from the group of 0, 1, and 2;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0169] Further separate embodiments provide a compound, respectively, of Formula (CXV), Formula (CXVI), Formula (CXVII), Formula (CXVIII), Formula (CXVIX), Formula (CXX), and Formula (CXXI), wherein, in each separate embodiment:Rs is selected from the group of C1-C4 alkyl and a moiety of the formula -(CH2)ni-O- (CH2)n2-CH3;nl is an integer independently selected from the group of 1 and 2; andn2 is an integer independently selected from the group of 0 and 1;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0170] Further separate embodiments provide a compound, respectively, of Formula (CXV), Formula (CXVI), Formula (CXVII). Formula (CXVIII), Formula (CXVIX), Formula (CXX), and Formula (CXXI), wherein, in each separate embodiment:R3 is selected from the group of C1-C3 alkyd and a moiety of the formula (CH2)ni-O- (CH2)n2-CH3;nl is an integer independently selected from the group of 1 and 2; andn2 is an integer independently selected from the group of 0 and 1;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0171] Further separate embodiments provide a compound, respectively, of Formula (CXV), Formula (CXVI), Formula (CXVII), Formula (CXVIII), Formula (CXVIX), Formula (CXX), and Formula (CXXI), wherein, in each separate embodiment:Rs is selected from the group of C1-C2 alkyl and a moiety of the formula -(CH2)ni-O- (CH2)n2-CHs;nl is 2; andn2 is 0;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0172] Further separate embodiments provide a compound, respectively, of each of Formula (CXXII), Formula (CXXIII), Formula (CXXIV), Formula (CXXV), and Formula (CXXVI):wherein, in each separate embodiment:Rs is selected from the group of Ci-Ce alkyl and a group of the formula -(CH2)ni-O-(CH2)n2-CH3;R.4 is selected from the group of Ci-Ce alkyl and a group of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; with the proviso that, when Rs is Ci-Ce alkyl, then R4 is Ci-Ce alkyl; and when Rs is (CH2)ni-O-(CH2)n2-CH3, then R4is Rs -(CH2)ni-O-(CH2)n2-CH3; andwith the proviso that the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH3moiety may be the same or different from the integers of nl and n2 in the Rs -(CH2)ni-O-(CH2)n2-CHs moiety;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0173] Additional separate embodiments provide a compound, respectively, of each of Formula (CXXII), Formula (CXXIII), Formula (CXXIV), Formula (CXXV), and Formula (CXXVI), wherein, in each separate embodiment:Rs is selected from the group of C1-C4 alkyl and a group of the formula -(CH2)ni-O-(CH2)n2-CHs;R4 is selected from the group of C1-C4 alkyl and a group of the formula -(CH2)ni-O-(CH2)n2-CHs;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; with the proviso that, when Rs is C1-C4 alkyl, then R4 is C1-C4 alkyl; and when Rs is -(CH2)ni-O-(CH2)n2-CHs, then R4is Rs -(CH2)ni-O-(CH2)n2-CH3; andwith the proviso that the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CHs moiety may be the same or different from the integers of nl and n2 in the Rs -(CH2)ni-O-(CH2)n2-CHs moiety;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0174] Additional separate embodiments provide a compound, respectively, of each of Formula ((CXXII), Formula (CXXIII), Formula (CXXIV), Formula (CXXV), and Formula (CXXVI), wherein, in each separate embodiment: Rs is Ci-Ce alkyl; and R4 is Ci-Ce alkyl; or apharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0175] Additional separate embodiments provide a compound, respectively, of each of Formula (CXXII). Formula (CXXIII), Formula (CXXIV), Formula (CXXV), and Formula (CXXVI). wherein, in each separate embodiment: Rs is C1-C4 alkyl; and R4 is C1-C4 alkyl; or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0176] Additional separate embodiments provide a compound, respectively, of each of Formula (CXXII), Formula (CXXIII), Formula (CXXIV), Formula (CXXV), and Formula (CXXVI), wherein, in each separate embodiment: Rs is C1-C3 alkyl; and R4 is C1-C3 alkyl; or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0177] Additional separate embodiments provide a compound, respectively, of each of Formula (CXXII), Formula (CXXIII), Formula (CXXIV), Formula (CXXV), and Formula (CXXVI), wherein, in each separate embodiment: Rs is C1-C2 alkyl; and R4 is C1-C2 alkyl; or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0178] Additional separate embodiments provide a compound, respectively, of each of Formula (CXXII), Formula (CXXIII), Formula (CXXIV), Formula (CXXV), and Formula (CXXVI), wherein, in each separate embodiment:Rs is a group of the formula -(CH2)ni-O-(CH2)n2-CH3;R4 is a group of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2: with the proviso that the integers of nl and n2 in the R3 -(CH2)ni-O-(CH2)n2-CH3 moiety may be the same or different from the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH3 moiety;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0179] Further separate embodiments provide a compound, respectively, of each of Formula (CXXII), Formula (CXXIII), Formula (CXXIV), Formula (CXXV), and Formula (CXXVI), wherein, in each separate embodiment:R3 is a group of the formula -(CH2)ni-O-(CH2)n2-CH3;R4 is a group of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1 and 2; n2 in each instance is an integer independently selected from the group of 0 and 1; with the proviso that the integers of nl and n2 in the R3 -(CH2)ni-O-(CH2)n2-CH3 moiety may be the same or different from the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH3 moiety;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0180] Further separate embodiments provide a compound, respectively, of each of Formula (CXXVII) and Formula (CXXVIII):wherein, in each separate embodiment:R3 is selected from the group of Ci-Ce alkyl and a group of the formula (CH2)ni-O-(CH2)n2-CH3;R4 is selected from the group of Ci-Ce alkyl and a group of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; R7 is selected from the group of methoxy and ethoxy; andRs, when present, is selected from the group of H, methoxy and ethoxy;with the proviso that, when R3 is Ci-Ce alkyl, then R4 is Ci-Ce alkyl; and when R3 is -(CH2)ni-O-(CH2)n2-CH3, then R4is R3 -(CH2)ni-O-(CH2)n2-CH3; andwith the proviso that the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH3 moiety may be the same or different from the integers of nl and n2 in the R3 -(CH2)ni-O-(CH2)n2-CH3 moiety;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0181] Further separate embodiments provide a compound, respectively, of each of Formula (CXXVII) and Formula (CXXVIII). wherein:R3 is selected from the group of C1-C4 alkyl and a group of the formula -(CH2)ni-O-(CH2)n2-CH3;R4 is selected from the group of C1-C4 alkyl and a group of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; R7 is selected from the group of methoxy and ethoxy; andRs, when present, is selected from the group of H, methoxy and ethoxy;with the proviso that, when R3 is C1-C4 alkyl, then R4 is C1-C4 alkyl; and when Rs is -(CH2)ni-O-(CH2)n2-CH3, then R4is R3 -(CH2)ni-O-(CH2)n2-CH3; andwith the proviso that the integers of nl and n2 in the R4(CH2)ni-O-(CH2)n2-CHs moiety' may be the same or different from the integers of nl and n2 in the Rs -(CH2)ni-O-(CH2)n2-CH3 moiety;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0182] Further separate embodiments provide a compound, respectively, of each of Formula (CXXVII) and Formula (CXXVIII), wherein, in each separate embodiment: Rs is Ci-Ce alkyl; R4is Ci-Ce alkyl; R7 is selected from the group of methoxy^ and ethoxy; and Rs, when present, is selected from the group of H, methoxy and ethoxy; or a pharmaceutically acceptable salt, cocrystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0183] Further separate embodiments provide a compound, respectively, of each of Formula (CXXVII) and Formula (CXXVIII), wherein, in each separate embodiment: R3 is C1-C4 alkyl; R4is C1-C4 alkyl; R7 is selected from the group of methoxy and ethoxy; and Rs, when present, is selected from the group of H, methoxy’ and ethoxy; or a pharmaceutically acceptable salt, cocrystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0184] Further separate embodiments provide a compound, respectively, of each of Formula (CXXVII) and Formula (CXXVIII), wherein, in each separate embodiment: R3 is C1-C3 alkyl; R4 is C1-C3 alkyl; R7 is selected from the group of methoxy and ethoxy; and Rs, when present, is selected from the group of H, methoxy and ethoxy; or a pharmaceutically acceptable salt, cocrystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0185] Further separate embodiments provide a compound, respectively, of each of Formula (CXXVII) and Formula (CXXVIII), wherein, in each separate embodiment: R3 is C1-C2 alkyl; R4 is C1-C2 alkyl; R7 is selected from the group of methoxy and ethoxy; and Rs, when present, is selected from the group of H, methoxy and ethoxy; or a pharmaceutically acceptable salt, cocrystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0186] Further separate embodiments provide a compound, respectively, of each of Formula (CXXVII) and Formula (CXXVIII), wherein, in each separate embodiment:R3 is a group of the formula -(CH2)ni-O-(CH2)n2-CH3;R4 is a group of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; R7 is selected from the group of methoxy and ethoxy; andRs, when present, is selected from the group of H, methoxy and ethoxy;with the proviso that the integers of nl and n2 in the R3 -(CH2)ni-O-(CH2)n2-CH3 moiety may be the same or different from the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH moiety;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0187] Further separate embodiments provide a compound, respectively, of each of Formula (CXXVII) and Formula (CXXVIII), wherein, in each separate embodiment:R3 is a group of the formula -(CH2)ni-O-(CH2)n2-CHs;R4 is a group of the formula -(CH2)ni-O-(CH2)n2-CH3;R7 is selected from the group of methoxy and ethoxy; andRs, when present, is selected from the group of H, methoxy and ethoxy;nl in each instance is an integer independently selected from the group of 1 and 2; n2 in each instance is an integer independently selected from the group of 0 and 1;with the proviso that the integers of nl and n2 in the R3 -(CH2)ni-O-(CH2)n2-CH? moiety may be the same or different from the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CHs moiety;or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0188] For each of the previous embodiments herein providing a compound of Formula (CXXVII), there is an additional embodiment providing a compound of Formula (CXXVII) in which R3 and R4 (including nl and n2, when present) are as defined in the previous embodiment in question and R7 is methoxy; or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0189] For each of the previous embodiments herein providing a compound of Formula (CXXVII), there is an additional embodiment providing a compound of Formula (CXXVII) in which R3 and R4 (including nl and n2, when present) are as defined in the previous embodiment in question and R7 is methoxy bound to the 4-position carbon atom of the azetidinyl ring; or a pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0190] For each of the previous embodiments herein providing a compound of Formula (CXXVIII), there is an additional embodiment providing a compound of Formula (CXXVIII) in which R3 and R4 (including nl and n2, when present) are as defined in the previous embodiment in question; R7 is methoxy; and Rs is methoxy; or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereol), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0191] For each of the previous embodiments herein providing a compound of Formula (CXXVIII), there is an additional embodiment providing a compound of Formula (CXXVIII) in which Rs and R4 (including nl and n2, when present) are as defined in the previous embodiment in question; R7 is methoxy bound to the 3-position carbon atom on the pyrrolidinyl ring; and Rs is methoxy bound to the 4-position carbon atom on the pyrrolidinyl ring; or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

[0192] In some embodiments, the nerve homing agent homing agent is selected from:

[0193] In some embodiments, the nerve homing agent homing agent is detected at a 500-700 nm excitation.

[0194] In some embodiments, the nerve homing agent homing agent is detected at a 625-800 nm emission.

[0195] In some embodiments, the nerve homing agent homing agent is:

[0196] In some embodiments, the nerve homing agent homing agent is detected at a 650 nm peak excitation.

[0197] In some embodiments, the nerve homing agent homing agent is detected at a 676 nm peak emission.

[0198] In some embodiments, the nerve homing agent is a molecule of Formula (DI):Rs is hydrogen or R2and R3 together form a fused ring, creating a core of FormulaR4 and Rs, together with the nitrogen atom to which they are bound, form a ring selected from the group of:or, when the compound is of Formula (DII), R4 and Rs may be independently selected from Ci-Ce alkyl, with the proviso that, when R4 is ethyl, Rs is not ethyl; Re is hydrogen;or, when R2 and R3 together form a fused ring to create a core of Formula (Dill), Rs and Re may also, together with the nitrogen atom to which Rs is bound, form afused ring, creating a core of Formula (Dill):with the proviso that, when R4 and Rs, together with the nitrogen atom to which theyare bound, form the ring, and R3 is H, Ri and R2, together with the nitrogen atom to which they are bound, may form a pyrrolidinyl ring;Xi in each instance is independently selected from Ci-Ce straight or branched alkyl, Ci-Ce straight or branched alkenyl, Ci-Ce straight or branched alkynyl, and - Si(Ci-C4alkyl)3;n is an integer selected from the group of 1 and 2;nl is an integer independently selected in each instance from the group of 1, 2, 3. and 4;n2 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;n3 is an integer independently selected in each instance from the group of 1, 2, 3. 4, 5, 6, 7, 8, 9, and 10;n4 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; andwith the proviso that the sum of n2 + n2 is not greater than 10;with the proviso that the sum of n2 + n3 is not greater than 10;with the proviso that the sum of n2 + n4 is not greater than 10;with the proviso that the sum of n3 + n4 is not greater than 10;with the proviso that, when R4 and Rs, together with the nitrogen atom to which theyare bound, form the ring, Ri and R2 are not both methyl, Ri and R2 arenot both ethyl, Ri and R2 are not both n-propyl, Ri and R2 are not both n-butyl, and Ri and R2 are not both n-pentyl;with the proviso that, when the compound is of Formula (DI), when Ri is methyl, R4 is not methyl; andwith the proviso that, when the compound is of Formula (DI), when Ri is ethyl, R4 is not ethyl.

[0199] In some embodiments, the nerve homing agent homing agent is selected from:

[0200] In some embodiments, the nerve homing agent homing agent is detected at a 500-725 nm excitation.

[0201] In some embodiments, the nerve homing agent homing agent is detected at a 600-800 nm emission.

[0202] In some embodiments, the nerve homing agent homing agent is:

[0203] In some embodiments, the nerve homing agent homing agent is detected at a 655 nm peak excitation.

[0204] In some embodiments, the nerve homing agent homing agent is detected at a 678 nm peak emission.

[0205] In some embodiments, the nerve homing agent is a molecule of Formula (DIV):Wherein:R1is Ci-6 alkyl.

[0206] In some embodiments, the nerve homing agent homing agent is:

[0207] In some embodiments, the nerve homing agent is a molecule of Formula (El):wherein:R1is selected from the group of -Xi, -(CH2)ni-SO3‘, -(CH2)ni-N (CH3)v -CH2-CH2- O-Xi, -CH2-CH2-O-[CH2-CH2-O]n4-Xi, -CH2-CH2-CH2-O-X1, and -CH2-CH2- CH2-O-[CH2-CH2-CH2-O]n5-Xl,Xi in each instance is independently selected from the group of Ci-Ce straight or branched alky l, C2-C6 straight or branched alkenyl, Ci-Ce straight or branched alkynyl, and -Si(Ci-C4 alkyl)3;nl is an integer independently selected in each instance from the group of 1, 2, 3. and 4;n2 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;n3 is an integer independently selected in each instance from the group of 1, 2, 3. 4, 5, 6, 7, 8, 9, and 10;n4 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; andn5 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5. 6, 7, 8. 9, and 10;with the proviso that the sum of n2 + n4 is not greater than 10;with the proviso that the sum of n3 + n5 is not greater than 10;with the proviso that the sum of n2 + n5 is not greater than 10; andwith the proviso that the sum of n3 + n4 is not greater than 10.

[0208] In some embodiments, the nerve homing agent homing agent is selected from:

[0209] In some embodiments, the nerve homing agent homing agent is detected at a 450-700 nm excitation.

[0210] In some embodiments, the nerve homing agent homing agent is detected at a 600-800 nm emission.

[0211] In some embodiments, the nerve homing agent is a molecule of Formula (FI):wherein:one of R1and R2is hydrogen;the other of R1and R2is selected from the group of:R3and R4are independently selected from the group of H, -CN. -O-Ci-Ce alkyl, — NH2, NH(CI-C6alkyl), — N(CI-C6alkyl)2, — CH=O, and — SO2(Ci-C6alkyl); with the proviso that not more than one of R3 and R4 is hydrogen.

[0212] In some embodiments, the nerve homing agent homing agent is selected from:

[0213] In some embodiments, the nerve homing agent homing agent is detected at a 450-700 nm excitation.

[0214] In some embodiments, the nerve homing agent homing agent is detected at a 600-800 nm emission.

[0215] In some embodiments, the nerve homing agent is a molecule of Formula (GI):R1and R2are each independently selected from hydrogen and C1-C4 alkyl; R3aand R3bare independently selected from hydrogen and C1-C4 alkyl, with the proviso that at least one of R3aand R3bis not hydrogen; or R3aand R3btogether with the nitrogen atom to which they are bound form a 5- or 6-membered nitrogen-containing ring;or R3bis selected from hydrogen and C1-C4 alkyl, and R3atogether with the nitrogen to which it is bound and R4forms a fused 5-membered or 6-membered ring, the 5-membered or 6-membered ring having one nitrogen heteroatom, two nitrogenheteroatoms, or one nitrogen heteroatom and one oxygen heteroatom, wherein each nitrogen heteroatom is optionally substituted by a C1-C4 alkyl substituent; or R3a, the nitrogen to which it is bound, and R4form a fused 6-membered, nitrogencontaining ring and R3b, the nitrogen to which it is bound, and R2together form a fused 6-membered, nitrogen-containing ring;R4and R7are each independently selected from hydrogen, halogen, or C1-C4 alkyl; R5and R6are each independently selected from hydrogen or C1-C4 alkyl; R8aand R8bare independently selected from hydrogen and C1-C4 alkyl, with the proviso that at least one of R8aand R8bis not hydrogen; or R8aand R8btogether with the nitrogen to which they are bound form a 5- or 6-membered nitrogencontaining ring optionally substituted by 1, 2, 3, or 4 C1-C4 alkyl substituents; or R8bis selected from hydrogen and C1-C4 alkyl, and R8atogether with the nitrogen to which it is bound and R7forms a fused 6-membered ring having one nitrogen heteroatom, two nitrogen heteroatoms, or one nitrogen heteroatom and one oxygen heteroatom, wherein each nitrogen heteroatom is optionally substituted by 1, 2, 3, or 4 C1-C4alkyl substituents;or R8a, the nitrogen to which it is bound, and R7form a fused 6-membered, nitrogencontaining ring and R8b, the nitrogen to which it is bound, and R1together form a fused 6-membered, nitrogen-containing ring;with the proviso that the compounds of Formula I do not include N3, N3, N7, N7- tetraethyl-10H-phenoxazine-3,7-diamine; N3, N7-di ethyl- 1 OH-phenoxazine-3,7 - diamine; N3, N3, N7, N7-tetramethyl-10H-phenoxazine-3,7-diamine; and N7, N7- diethyl-N3, N3,2-trimethyl-10H-phenoxazine-3,7-diamine.

[0216] In some embodiments, the nerve homing agent homing agent is selected from:

[0217] In some embodiments, the nerve homing agent homing agent is detected at a 450-700 nm excitation.

[0218] In some embodiments, the nerve homing agent homing agent is detected at a 550-800 nm emission.

[0219] In some embodiments, the nerve homing agent is a molecule of Formula (HI):wherein:R is a straight or branched alkyl chain of from 2 to 12 carbon atoms;R1is selected from the group of methyl, ethyl, n-propyl, isopropyl, — (CH2)ni — SO3, — (CH2)m— N+(CH3)3, — CH2— CH2— O— XI, — CH2— CH2— O— [CH2— CH2— O]n2— XI, — CH2— CH2— CH2— O— XI, and —CH2—CH2— CH2— O— [CH2— CH2— CH2— O]n3— Xi,Xi in each instance is independently selected from Ci-Ce straight or branched alkyl, C2-C6straight or branched alkenyl, Ci-Ce straight or branched alkynyl, and — Si(Ci-C4alkyl)3;nl is an integer independently selected in each instance from the group of 1, 2, 3, and 4;n2 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;n3 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9. and 10;n4 is an integer independently selected in each instance from the group of 1, 2, 3. 4, 5, 6, 7, 8, 9, and 10; andn5 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9. and 10;with the proviso that the sum of n2+n4 is not greater than 10;with the proviso that the sum of n3+n5 is not greater than 10;with the proviso that the sum of n2+n5 is not greater than 10; andwith the proviso that the sum of n3+n4 is not greater than 10.

[0220] In some embodiments, the nerve homing agent homing agent is selected from:

[0221] In some embodiments, the nerve homing agent homing agent is detected at a 450-700 nm excitation.

[0222] In some embodiments, the nerve homing agent homing agent is detected at a 600-800 nm emission.

[0223] In some embodiments, the nerve homing agent is a molecule of Formula (JI):wherein:R1is selected from hydrogen and C1-C3 alkyl;R2is selected from hydrogen and C1-C3 alkyl;R3is selected from the group of hydrogen, halogen, and C1-C3 alkyl;or, when R3is bound to the 9-position carbon of the 10H-benzo[c]phenoxazine core, R2and R3, together with the carbon atoms to which they are bonded, form a fivemembered or six-membered fused ring, the five-membered or six-membered fused ring optionally containing a ring oxygen heteroatom, and the five-membered or six-membered fused ring being substituted by 0, 1, 2, or 3 Ci- C3 alkyl substituents; andR4and R5are each independently selected from hydrogen and C1-C3 alkyl.

[0224] In some embodiments, the nerve homing agent homing agent is selected from:

[0225] In some embodiments, the nerve homing agent homing agent is detected at a 450-800 nm excitation.

[0226] In some embodiments, the nerve homing agent homing agent is detected at a 550-850 nm emission.

[0227] In some embodiments, the tumor homing agent and the nerve homing agent each detect a physiologic structure or cell between at least 10 pm and 25 pm in diameter.

[0228] In some embodiments, the nerve homing agent detects a physiologic structure or cell between at least 10 pm and 25 pm in diameter.

[0229] Some embodiments further comprise causing remission in, reducing, ameliorating, or ablating the cancer or vascular lesion, include remission, increasing quality of life, reducing the extent of resection, extending progression free survival, or number of surgeries required, reducing pain, or exhibiting other clinical or quality of life benefit to patent.BRIEF DESCRIPTION OF THE DRAWINGS

[0230] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings.

[0231] FIG. 1 illustrates representative examples of coadministration and detection of a nerve specific and a tumor specific agent in a head and neck cancer model, across two mock HNSCC resections (the first in row 1 and the second in row 2 of FIG. 1) in a Detroit-562 mouse model. White light images shown in FIG. 1A and FIG. IB respectively. The NIR fluorescence emission of the nerve-homing Compound C9 agent was shown in nerve tissue when excited at a broad fluorescence wavelength of 590-650 nm (denoted as Compound C9, FIG. 1C and FIG. ID respectively). The NIR fluorescence emission of the tumor homing agent was shown in the tumor tissue when excited at a broad fluorescence wavelength of 672.5-747.5 nm (denoted as Compound 76, FIG. IE and FIG. IF respectively). The nerve-specific labeling of Compound C9 and tumor-specific labeling of the peptide-fluorophore complex, Compound 76 containing a peptide of SEQ ID NO: 9 complexed to an ICG, respectively relative to nerve and tumor structures seen under white light (FIG. 1A and FIG. IB respectively) clearly illuminated the nerve and tumor anatomical structures respectively in the HNSCC tumor. Moreover, as shown in FIG. 1G and FIG. 1H, the Compound C9 nerve homing agent, based on the acquired andquantified fluorescence intensity at applicable wavelength of 662.5-737.5 nm, localized significantly and specifically in nerve relative to both nearby muscle (background) and tumor signals shown in FIG. 1G. Complimentarily, based on the acquired and quantified fluorescence intensity at applicable wavelength of >780 nm, the Compound 76 tumor-homing agent compound localized significantly and specifically in tumor relative to nearby muscle (background) and nerve signal (Compound C9 fluorescent agent) as shown in FIG. 1H. These results showed that in the Detroit-562 model, the selective nerve homing agents and tumor homing agents in fact respectively homed to the appropriate tissue and cellular structures in the model and their fluorescence signals were distinct and detected in the same samples without cross talk.

[0232] FIG. 2 illustrates additional representative examples of coadministration and detection of a nerve specific and a tumor specific agent in another head and neck cancer model, across two mock HNSCC resections (the first in row 1 and the second in row 2 of FIG. 2) in a FaDu HNSCC mouse model. White light images shown in FIG. 2A and FIG. 2B respectively. Using the methods described in FIG. 1, the samples were excited at a broad fluorescence w avelength as depicted. The NIR fluorescence emission of the nerve-homing Compound C9 agent w as shown in nerve tissue the fluorescence wavelength of 662.5-737.5 nm (denoted as Compound C9. FIG. 2C and FIG.2D respectively). The NIR fluorescence emission of the tumor homing agent (Compound 76) was shown in the tumor tissue the fluorescence wavelength of >780 nm (denoted as Compound 76, FIG. 2E and FIG. 2F respectively). The nerve specific labeling of Compound C9 and the tumor-specific labeling of the peptide-fluorophore complex (Compound 76) (containing a peptide of SEQ ID NO: 9 complexed to an ICG), respectively relative to nerve and tumor structures seen under white light (FIG. 2A and FIG. 2B respectively) clearly illuminated the nerve and tumor anatomical structures in the HNSCC tumor. Moreover, as shown in FIG. 2G and FIG. 2H, the Compound C9 nerve homing agent based on the acquired and quantified fluorescence intensity at applicable wavelength of 662.5-737.5 nm localized significantly and specifically in nerve relative to both nearby muscle (background) and tumor signals shown in FIG. 2G. Complimentarily, based on the acquired and quantified fluorescence intensity at applicable wavelength of >780 nm the Compound 76 tumor-homing agent localized significantly and specifically in tumor relative to nearby muscle (background) and nerve signal (Compound C9 fluorescent agent) as shown in FIG. 2H. These results showed that in an additional model, the FaDu HNSCC model, the selective nerve and tumor homing agents in fact homed to the appropriate structures in the model and their fluorescence signals w ere distinct and detected in the same samples without cross talk.

[0233] FIG. 3 illustrates examples of administration and detection of a nerve specific agent only in a head and neck cancer model compared across different fluorescent emission spectra, across two mock HNSCC resections (the first in row 1 and the second in row 2 of FIG. 3) in a FaDu HNSCC mouse model using only the fluorescence excitation and emission wavelengths of 590-650 nm and 662.5-737.5, respectively to determine whether any tumor structures were also detected at such wavelength in the same sample. White light images are shown in FIG. 3A and FIG. 3B respectively. The NIR fluorescence emission of the nerve-homing Compound C9 agent was shown in nerve tissue when excited at a broad fluorescence wavelength of 662.5-737.5 nm (denoted as Compound C9 in FIG. 3C and FIG. 3D respectively). The NIR fluorescence emission of the nerve-homing Compound C9 agent was not detectable in any tissue in the sample (FIG. 3E and FIG. 3F respectively) in the channel with the NIR fluorescence excitation and emission of 672.5-747.5 nm and >780 nm, respectively, corresponding to the tumor homing agent Compound 76. This result shows the absence of Compound C9 signal crosstalk into the Compound 76 fluorescence channel (FIG. 3E and FIG. 3F). Moreover, this result is supported by the quantified localized fluorescence intensity of Compound C9 nerve homing agent, in the nerve homing agent fluorescence channel of 662.5-737.5 nm, localized significantly and specifically in nerve relative to nearby muscle and tumor signal as shown in FIG. 3G, and was not detected at all in the Compound 76 fluorescence channel of >780 nm as shown in FIG. 3H.These results showed that Compound C9 nen e agent exhibited no crosstalk or detectability under the emission spectrum measured for the Compound 76 fluorophore. This result shows that the emission detected in the sample for the Compound C9 nerve agent is specific for that agent and is not confused with a signal under the different wavelength tailored to the tumor-homing agent.

[0234] FIG. 4 illustrates further examples of administration and detection of a tumor specific agent only in a head and neck cancer model compared across different fluorescent emission spectra, across two mock HNSCC resections (the first in row 1 and the second in row 2 of FIG.4) in a Detroit-562 HNSCC mouse model using only the fluorescence excitation and emission wavelengths when excited at a broad fluorescence wavelength of 672.5-747.5 nm and >780 nm, respectively, to determine whether any nerve structures was also detected at such wavelength in the same sample. White light images are shown in FIG. 4A and FIG. 4B. The NIR fluorescence emission of the tumor-homing Compound 76 agent was shown in tumor tissue at the fluorescence wavelength of the agent of >780 nm (denoted as Compound 76, FIG. 4E and FIG.4F respectively) in the channel with the NIR fluorescence emission of >780 nm that corresponds with that of the tumor homing agent. The NIR fluorescence emission of the tumor-homing agent Compound 76 was not detectable in any tissue in the sample (FIG. 4C and FIG.4D respectively) in the channel with the NIR fluorescence excitation and emission of 590-650 nm and 662.5-737.5 nm, respectively corresponding to the nerve homing agent D12. This result shows the absence of Compound 76 signal crosstalk into the Compound D12 fluorescence channel (FIG. 4C and FIG. 4D). Moreover, this result is supported by the quantified localized fluorescence intensity of Compound 76 tumor homing agent, in the tumor homing agent fluorescence channel of >780 nm, localized significantly and specifically in tumor relative to nearby muscle and nerve signal as shown in FIG.4H and was not detected at all in the Compound D12 fluorescence channel of 662.5-737.5 nm as shown in FIG.4G. These results showed that Compound 76 tumor agent exhibited no crosstalk or detectability under the emission spectrum measured for the Compound D12 fluorophore. This result shows that the emission detected in the sample for the Compound 76 tumor agent is specific for that agent and is not confused with a signal under the different wavelength tailored to the nerve-homing agent.

[0235] FIG. 5 illustrates representative examples of coadministration and detection of a nerve specific and a tumor specific agent in a head and neck cancer model, across two mock HNSCC resections (the first in row 1 and the second in row 2 of FIG.5) in a Detroit-562 mouse model. White light images are shown in FIG. 5A and FIG. 5B. Using the methods shown in FIG. 1, the samples were excited at a broad fluorescence wavelength as depicted. The NIR fluorescence emission of the nerve-homing Compound D12 agent was shown in nerve tissue when excited at the fluorescence wavelength of 590-650 nm (denoted as Compound D12, FIG. 5C and FIG. 5D respectively). The NIR fluorescence emission of the tumor homing agent, Compound 76, w as shown in the tumor tissue when excited at the fluorescence wavelength of 672.5-747.5 nm (denoted as Compound 76, FIG. 5E and FIG. 5F respectively). The nerve-specific labeling of Compound D12 and tumor-specific labeling of the peptide-fluorophore complex, Compound 76 (containing a peptide of SEQ ID NO: 9 complexed to an ICG), respectively relative to nerve and tumor structures seen under white light (FIG. 5A and FIG.5B respectively) clearly illuminated the nerve and tumor anatomical structures respectively in the HNSCC tumor. Moreover, as shown in FIG. 5G and FIG. 5H, the Compound D12 nerve homing agent based on the acquired and quantified fluorescence intensity at applicable wavelength of 662.5-737.5 nm localized significantly and specifically in nerve relative to both nearby muscle (background) and tumor signals shown in FIG. 5G. Complimentarily, based on the acquired and quantified fluorescence intensity at applicable wavelength of >780 nm the Compound 76 tumor-homing agent compound localized significantly and specifically in tumor relative to nearby muscle (background) and nerve signal (Compound D12 fluorescent agent) as shown in FIG. 5H. Theseresults showed that in the Detroit-562 model, the selective nerve homing agents and tumor homing agents in fact respectively homed to the appropriate tissue and cellular structures in the model and their fluorescence signals were distinct and detected in the same samples without cross talk.

[0236] FIG. 6 illustrates nerve-specificity screening in mice for nerve Compounds C1-C13 where all compounds and derivates demonstrated nerve uptake, with 12 of the 13 library compounds demonstrating positive nerve signal to background ratios (SBR).

[0237] FIG. 7 illustrates a subset of 7 exemplary compounds from the C1-C13 library that were additionally screened in rat nerve models of the peripheral and central nervous systems, with comparative images of DI 2 included for reference. All compounds demonstrated strong PNS contrast in the brachial plexus (top row images) and CNS contrast in the cranial nerves (2ndrow images) following systemic administration. Several derivates displayed excellent white matter tract (WMT) specific signal (3rdrow images) and a majority’ demonstrated contrast in facial nerves (4throw images).

[0238] FIG. 8 illustrates the nerve-specificity following systemic administration of the compound library. FIG.8A provides the chemical structures with representative white light and fluorescence images of the seven oxazine-based fluorophores with the brightest nerve tissue fluorescence intensity after systemic administration are shown for comparison to the parent G21 (7) and a vehicle-injected control group. FIG. 8B provides quantified nerve, muscle, and adipose tissue intensity’ values reported as A. U. / s following direct fluorophore administration. All fluorophores were systemically administered at 7.5 pmol / kg or equivalent volume of blank cosolvent formulation. All images represent data collected across n = 4 mice (50 / 50, male / female). Images are shown with optimal contrast. Relative intensity values, expressed in decimal form, are shown boxed in the lower left-hand comer representing the measured value of each compound imaged relative to the brightest fluorophore, D14 (24). FIG. 8C provides quantified N / M SBRs of the 28 oxazine-based derivatives and vehicle-injected control group following systemic administration are arranged in ascending order. All values are presented as the mean ± SD. The seven highlighted oxazine-based compounds are equivalently color-coded throughout the figure. Ctrl = vehicle-injected control to quantity’ autofluorescence. FL = Fluorescence. Data for each fluorophore were compared to the vehicle-injected control group to evaluate statistical significance of nerve SBR via one-way ANOVA followed by a Fisher’s LSD multiple comparison test with no assumption of sphericity' using the Geisser-Greenhouse correction, where *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. FIG.8D provides a scatter plot of the average nerve tissue fluorescence intensities of all 28 G21 derivatives andvehicle injected control group plotted against the average N / M SBR following systemic administration.

[0239] FIG. 9 illustrates the fluorescent buried nerve imaging at variable depths. FIG.9A provides white light (top) and fluorescence (bottom) images of murine brachial plexus nerves collected following systemic administration of 7.5 pmol / kg dose of G21. D12, D14 and D5. Images were collected of surgically exposed nerves and with the same region of interest (ROI) covered with 1 or 2 mm of muscle tissue layered over the exposed nerve. Rectangle dashed lines and arrows indicate the region and direction, respectively, of the brachial plexus nerve line profile measured in FIG. 9B. Square dashed lines indicate the boundaries of the overlaid muscle tissue. Oval / round dashed lines and arrows indicate the region and direction, respectively, of the brachial plexus nerve line profile measured in FIG.9C. FIG.9B provides cross-sectional line profiles of fluorescence intensity of D5, D12, D14, G21, and a vehicle injected control (Ctrl) quantified for the ROI representing small diameter nerves visualized at the 1-mm depth, indicated by the rectangle dashed lines in FIG. 9A. FIG. 9C provides Cross-sectional line profiles of fluorescence intensity of the same compounds and vehicle injected control quantified for the ROI representing large diameter nerves visualized at the 2-mm depth, indicated by the orval / round dashed lines in FIG. 9A. FIG. 9D provides three-dimensional topographical heat maps of the fluorescence intensity from D12 and D5 generated from the brachial plexus images simulating nerves buried beneath 1 and 2 mm, respectively, of muscle tissue. Color bars represent pixel intensity values in arbitrary units (A. U.). All data represented was collected for n = 2 mice per fluorophore. Ctrl = vehicle injected control to quantity' autofluorescence. FL = Fluorescence.

[0240] FIG. 10 illustrates the peak-normalized absorbance and fluorescence spectra of C9.

[0241] FIG. 11 illustrates the peak-normalized absorbance and fluorescence spectra of D12.DETAILED DESCRIPTION

[0242] Currently, no technology exists to simultaneously enhance intraoperative cancer and nerve recognition, to delineate tumor tissue from non-tumor tissue alongside nerves in the same sample or in vivo, and no technology' exists to combine the two (e.g., a tumor homing and nerve homing agent) to aid surgeons in a single intraoperative surgical setting. Thus, technology as described herein provides a direct visualization of tissue, cells, and margins and direct visualization of nerves simultaneously in in surgical settings and pathology samples. The compositions and methods as described herein would greatly improve surgical outcomes in cancer surgeries and reduce comorbidities for patients.

[0243] Fluorescence Guided Surgery (FGS) is currently being explored and has great potential for integrating into clinical medicine, with a prospect of allowing surgeons visualization of important cells, tissues, biological structures, and complex anatomy. FGS imaging systems are being developed to operate in the near-infrared (NIR, -650-900 nm) excitation range and other fluorescent emission ranges to visualize targeted fluorescent probes with high contrast in diseased tissue in situ, in an operating bed, or in tissue pathology samples.

[0244] Importantly, the tumor and nerve specific agents described herein are within the NIR spectrum, with tumor-targeting FGS probes described herein utilizing fluorescent reporters typically between the 600-900 nm wavelength ranges, while our nerve-specific probes fluoresce typically between the 600-775 nm wavelength ranges, providing spectrally distinct tissue detection and compatibility of each fluorescent agent with existing two-color FGS systems. The combination of these two promising technologies herein provides a comprehensive, innovative solution to enhance cancer control and removal and nerve sparing during cancer surgical resection.

[0245] Several FGS imaging systems capable of real-time open or laparoscopic NIR fluorescence visualization are utilized clinically, at least three of which are equipped with two-color imaging capabilities that would have direct utility for the inventions described herein, (e.g.. Quest Spectrum imaging systems (Quest). Fluobeam imaging systems (FluOptics). Lab Flare imaging systems (Curadel), or Solaris imaging systems (Perkin Elmer)).

[0246] The invention will best be understood by reference to the following detailed description of the aspects and embodiments of the invention, taken in conjunction with the accompanying drawings and figures. The discussion below is descriptive, illustrative and exemplary and is not to be taken as limiting the scope defined by any appended claims.

[0247] As used in the specification and appended claims, unless specified to the contrary, the following terms have the meaning indicated.

[0248] As used herein and in the appended claims, the singular forms “a,” "‘and,” and “‘the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a compound” includes a plurality of such compounds, reference to “an agent” includes a plurality of such agents, and reference to “the cell” includes reference to one or more cells (or to a plurality of cells) and equivalents thereof known to those skilled in the art, and so forth. When ranges are used herein for physical properties, such as molecular weight, or chemical properties, such as chemical formulae, all combinations and subcombinations of ranges and specific embodiments therein are intended to be included. The term “about” when referring to a number or a numerical range means that the number or numerical range referred to is anapproximation within experimental variability (or within statistical experimental error), and thus the number or numerical range may vary between 1% and 15% of the stated number or numerical range. The term “comprising” (and related terms such as “comprise” or “comprises” or “having” or “including”) is not intended to exclude that in other certain embodiments, for example, an embodiment of any composition of matter, composition, method, or process, or the like, described herein, may “consist of’ or “consist essentially of’ the described features.

[0249] “Cyano” refers to the -CN radical.

[0250] “Nitro” refers to the -NO2 radical.

[0251] “Oxa” refers to the -O- radical.

[0252] “Oxo” refers to the =0 radical.

[0253] “Thioxo” refers to the =S radical.

[0254] “Imino” refers to the =N-H radical.

[0255] “Hydrazine” refers to the =N-NH2 radical.

[0256] “Alkyd” refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to fifteen carbon atoms (e.g., C1-C15 alkyl). In certain embodiments, an alkyl comprises one to thirteen carbon atoms (e.g., C1-C13 alkyl). In certain embodiments, an alkyl comprises one to eight carbon atoms (e.g.. Ci-Cs alkyl). In other embodiments, an alkyl comprises five to fifteen carbon atoms (e.g., C5-C15 alky l). In other embodiments, an alkyl comprises five to eight carbon atoms (e g., Cs-Cs alkyl). The alkyl is attached to the rest of the molecule by a single bond, for example, methyl (Me), ethyl (Et), w-propyl, 1 -methylethyl (Ao-propyl), n-butyl, n-pentyl, 1,1 -dimethylethyl ( / -butyl). 3 -methylhexyl. 2-methylhexyl, and the like. Unless stated otherwise specifically in the specification, an alkyd group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, -ORa, - SR“, -OC(O)-Ra, -N(Ra)2, -C(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -N(Ra)C(O)Ra, -N (Ra)S(O)tRa(where t is 1 or 2), -S(O)tORa(where t is 1 or 2) and -S(O)tN(Ra)2 (where t is 1 or 2) where each Rais independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.

[0257] “Alkenyl” refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond, and having from two to twelve carbon atoms. In certain embodiments, an alkenyl comprises two to eight carbon atoms. In other embodiments, an alkenyl comprises two to four carbon atoms. The alkenyl is attached to the rest of the molecule by a single bond, for example, ethenyl (i.e., vinyl), prop-l-enyl (i.e., allyl), but-l-enyl, pent-l-enyl, penta- 1,4-dienyl, and the like. Unless statedotherwise specifically in the specification, an alkenyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, -ORa, -SRa, -OC(O)-Ra, -N(Ra)2, -C(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -N(Ra)C(O)Ra, -N (Ra)S(O)tRa(where t is 1 or 2), -S(O)tORa(where t is 1 or 2) and -S(O)tN(Ra)2 (where t is 1 or 2) where each Rais independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.

[0258] “Alkynyl” refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one triple bond, having from two to twelve carbon atoms. In certain embodiments, an alkynyl comprises two to eight carbon atoms. In other embodiments, an alkynyl has two to four carbon atoms. The alkynyl is attached to the rest of the molecule by a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unless stated otherwise specifically in the specification, an alkynyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, -ORa, - SRa, -OC(O)-Ra, -N(Ra)2, -C(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -N(Ra)C(O)Ra, -N (Ra)S(O)tRa(where t is 1 or 2), -S(O)tORa(where t is 1 or 2) and -S(O)tN(Ra)2 (where t is 1 or 2) where each Rais independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.

[0259] '‘Alkylene” or ‘'alkylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing no unsaturation and having from one to twelve carbon atoms, for example, methylene, ethylene, propylene, / 7-butylene. and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon in the alkylene chain or through any two carbons within the chain. Unless stated otherwise specifically in the specification, an alkylene chain is optionally substituted by one or more of the following substituents: halo, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, thioxo, trimethylsilanyl, -ORa, - SRa, -OC(O)-Ra, -N(Ra)2, -C(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -N(Ra)C(O)Ra, -N (Ra)S(O)tRa(where t is 1 or 2), -S(O)tORa(where t is 1 or 2) and -S(O)tN(Ra)2 (where t is 1 or 2) where each Rais independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.

[0260] “Alkenylene” or “alkenylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon andhydrogen, containing at least one double bond and having from two to twelve carbon atoms, for example, ethenylene, propenylene, w-butenylene, and the like. The alkenylene chain is attached to the rest of the molecule through a double bond or a single bond and to the radical group through a double bond or a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherw ise specifically in the specification, an alkenylene chain is optionally substituted by one or more of the following substituents: halo, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, thioxo, trimethylsilanyl, -ORa, - SRa, -OC(O)-Ra, -N(Ra)2, -C(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -N(Ra)C(O)Ra, -N (Ra)S(O)tRa(where t is 1 or 2), -S(O)tORa(where t is 1 or 2) and -S(O)tN(Ra)2 (where t is 1 or 2) where each Rais independently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkydalkyl, aryl (optionally substituted with one or more halo groups), aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, and where each of the above substituents is unsubstituted unless otherwise indicated.

[0261] “Aryl” refers to a radical derived from an aromatic monocyclic or multicyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom. The aromatic monocyclic or multicyclic hydrocarbon ring system contains only hydrogen and carbon from six to eighteen carbon atoms, where at least one of the rings in the ring system is fully unsaturated, 1.e., it contains a cyclic, delocalized (4n+2) 7t-electron system in accordance with the Hiickel theory. Aryl groups include, but are not limited to, groups such as phenyl, fluorenyl, and naphthyl. Unless stated otherwise specifically in the specification, the term “ary l” or the prefix “ar-“ (such as in “aralkyl”) is meant to include an radicals optionally substituted by one or more substituents independently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, cyano, nitro, optionally substituted and, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substitutedheteroarylalkyl, -Rb-ORa, -Rb-OC(O)-Ra, -Rb-N(Ra)2, -Rb-C(O)Ra, -Rb-C(O)ORa, -Rb-C(O)N(Ra) 2, -Rb-O-Rc-C(O)N(Ra)2, -Rb-N(Ra)C(O)ORa, -Rb-N(Ra)C(O)Ra, -Rb-N(Ra)S(O)tRa(where t is 1 or 2). -Rb-S(O)tORa(where t is 1 or 2) and -Rb-S(O)tN(Ra)2 (where t is 1 or 2), where each Rais independently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl, and (optionally substituted with one or more halo groups), aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or hcteroar lalkyl. each Rbis independently a direct bond or a straight or branched alky lene oralkenylene chain, and Rcis a straight or branched alkylene or alkenylene chain, and where each of the above substituents is unsubstituted unless otherwise indicated.

[0262] “Aralkyl” refers to a radical of the formula -Rc-aryl where Rcis an alkylene chain as defined above, for example, benzyl, diphenylmethyl and the like. The alkylene chain part of the aralkyl radical is optionally substituted as described above for an alkylene chain. The aryl part of the aralkyl radical is optionally substituted as described above for an aryl group.

[0263] “Aralkenyl” refers to a radical of the formula -Rd-aryl where Rdis an alkenylene chain as defined above. The aryl part of the aralkenyl radical is optionally substituted as described above for an aryl group. The alkenylene chain part of the aralkenyl radical is optionally substituted as defined above for an alkenylene group.

[0264] “Aralkynyl” refers to a radical of the formula -Re-aryl, where Reis an alkynylene chain as defined above. The ary l part of the aralkynyl radical is optionally substituted as described above for an aryl group. The alkynylene chain part of the aralkynyl radical is optionally substituted as defined above for an alkynylene chain.

[0265] “Carbocyclyl” refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems, having from three to fifteen carbon atoms. In certain embodiments, a carbocyclyl comprises three to ten carbon atoms. In other embodiments, a carbocyclyl comprises five to seven carbon atoms. The carbocyclyl is attached to the rest of the molecule by a single bond. Carbocyclyl may be saturated, (i.e., containing single C-C bonds only) or unsaturated (i.e., containing one or more double bonds or triple bonds.) A fully saturated carbocyclyl radical is also referred to as “cycloalkyl.” Examples of monocyclic cycloalkyls include, e.g.. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. An unsaturated carbocyclyl is also referred to as “cycloalkenyl.” Examples of monocyclic cycloalkenyls include, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Polycyclic carbocyclyl radicals include, for example, adamantyl, norbomyl (i.e., bicyclo[2.2. l]heptanyl). norbomenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2. l]heptanyl, and the like. Unless otherwise stated specifically in the specification, the term “carbocyclyl” is meant to include carbocyclyl radicals that are optionally substituted by one or more substituents independently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted ary l, optionally substituted aralkyl, optionally substituted aralkenyl. optionally substituted aralkynyl. optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substitutedheteroarylalkyl, -Rb-ORa, -Rb-SRa, -Rb-OC(O)-Ra, -Rb-N(Ra)2, -Rb-C(O)Ra, -Rb-C(O)ORa, -Rb-C (O)N(Ra)2, -Rb-O-Rc-C(O)N(Ra)2, -Rb-N(Ra)C(O)ORa, -Rb-N(Ra)C(O)Ra, -Rb-N(Ra)S(O)tRa(where t is 1 or 2), -Rb-S(O)tORa(where t is 1 or 2) and -Rb-S(O)tN(Ra)2 (where t is 1 or 2), where each Rais independently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, each Rbis independently a direct bond or a straight or branched alkylene or alkenylene chain, and Rcis a straight or branched alkylene or alkenylene chain, and where each of the above substituents is unsubstituted unless otherwise indicated.

[0266] “Carbocyclylalkyr refers to a radical of the formula -Rc-carbocyclyl where Rcis an alkylene chain as defined above. The alkylene chain and the carbocyclyl radical is optionally substituted as defined above.

[0267] “Halo” or “halogen” refers to bromo, chloro, fluoro or iodo substituents.

[0268] “Fluoroalkyl” refers to an alkyl radical, as defined above, that is substituted by one or more fluoro radicals, as defined above, for example, trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl, l-fluoromethyl-2-fluoroethyl, and the like. The alkyl part of the fluoroalkyl radical is optionally substituted as defined above for an alkyl group.

[0269] “Heterocyclyl” refers to a 3- to 18-membered non-aromatic ring radical that comprises two to twelve carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. Unless stated otherwise specifically in the specification, the heterocyclyl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems. The heteroatoms in the heterocyclyl radical may be optionally oxidized. One or more nitrogen atoms, if present, are optionally quatemized. The heterocyclyl radical is partially or fully saturated. The heterocyclyl may be attached to the rest of the molecule through any atom of the ring(s). Examples of such heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl. oxazolidinyl, piperidinyl, piperazinyl. 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in the specification, the term “heterocyclyl” is meant to include heterocyclyl radicals as defined above that are optionally substituted by one or more substituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl,optionally substituted carbocyclylalkyL optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -Rb-ORa, -Rb-SRa, -Rb-OC(O)-Ra, -Rb-N(Ra)2, -Rb-C(O)Ra, -Rb-C(O)ORa, -Rb-C (O)N(Ra)2, -Rb-O-Rc-C(O)N(Ra)2, -Rb-N(Ra)C(O)ORa, -Rb-N(Ra)C(O)Ra. -Rb-N(Ra)S(O)tRa(where t is 1 or 2), -Rb-S(O)tORa(where t is 1 or 2) and -Rb-S(O)tN(Ra)2 (where t is 1 or 2), where each Rais independently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalky l, heteroaryl or heteroarylalkyl, each Rbis independently a direct bond or a straight or branched alkylene or alkenylene chain, and Rcis a straight or branched alkylene or alkenylene chain, and where each of the above substituents is unsubstituted unless otherwise indicated.

[0270] ■■ / V-heterocyclyl" or "N-attached heterocyclyl” refers to a heterocyclyl radical as defined above containing at least one nitrogen and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a nitrogen atom in the heterocyclyl radical. An A'-heterocyclyl radical is optionally substituted as described above for heterocyclyl radicals. Examples of such / V-heterocyclyl radicals include, but are not limited to, 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1 -pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

[0271] “C-heterocyclyl” or “C-attached heterocyclyl” refers to a heterocyclyl radical as defined above containing at least one heteroatom and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a carbon atom in the heterocyclyl radical. A C-heterocyclyl radical is optionally substituted as described above for heterocyclyl radicals. Examples of such C-heterocyclyl radicals include, but are not limited to, 2-morpholinyl, 2- or 3-or 4-piperidinyl, 2-piperazinyl, 2- or 3-pyrrolidinyl, and the like.

[0272] ’‘Heterocyclylalkyl” refers to a radical of the formula -Rc-heterocyclyl where Rcis an alkylene chain as defined above. If the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl is optionally attached to the alkyl radical at the nitrogen atom. The alkylene chain of the heterocyclylalkyl radical is optionally substituted as defined above for an alkylene chain. The heterocyclyl part of the heterocyclylalkyl radical is optionally substituted as defined above for a heterocyclyl group.

[0273] “Heteroaryl” refers to a radical derived from a 3- to 18-membered aromatic ring radical that comprises two to seventeen carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. As used herein, the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, wherein at least one of the rings in the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2) 7t-electron system in accordance with the Huckel theory. Heteroaryl includes fused or bridged ring systems. The heteroatom(s) inthe heteroaryl radical is optionally oxidized. One or more nitrogen atoms, if present, are optionally quatemized. The heteroaryl is attached to the rest of the molecule through any atom of the ring(s). Examples of hctcroar ls include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[i>][l,4]dioxepinyl, benzo [b] [ 1.4] oxazinyl.1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[l,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6.7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[l,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,5.6.7.8.9.10-hexahydrocycloocta[d]pyridazinyl,5.6.7.8.9.10-hexahydrocycloocta|d]pyridinyl,isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl,5.8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl.1 -phenyl- 1 / 7-pyrrolyl, phenazinyl. phenothiazinyl, phenoxazinyl. phthalazinyl. pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5.6.7.8-tetrahydrobenzo[4.5]thieno[2,3-d]pyrimidinyl,6.7.8.9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e., thienyl). Unless stated otherwise specifically in the specification, the term "‘heteroaryT’ is meant to include heteroaryl radicals as defined above which are optionally substituted by one or more substituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl, haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -Rb-0Ra, -Rb-SRa, -Rb-0C(0)-Ra, -Rb-N(Ra)2, -Rb-C(0)Ra, -Rb-C(0)0Ra, -Rb-C (0)N(Ra)2, -Rb-0-Rc-C(0)N(Ra)2, -Rb-N(Ra)C(0)0Ra, -Rb-N(Ra)C(0)Ra, -Rb-N(Ra)S(O)tRa(where t is 1 or 2), -Rb-S(O)tORa(where t is 1 or 2) and -Rb-S(O)tN(Ra)2 (where t is 1 or 2), where each Rais independently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkydalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroary l or heteroary lalkyl, each Rbis independently a direct bond or a straight or branched alkylene or alkenylene chain, and Rcis a straight or branched alkylene or alkenylene chain, and where each of the above substituents is unsubstituted unless otherwise indicated.

[0274] ‘W-heteroary 1” refers to a heteroaryl radical as defined above containing at least one nitrogen and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a nitrogen atom in the heteroaryl radical. An A-heteroaryl radical is optionally substituted as described above for heteroaryl radicals.

[0275] “C-heteroaryd” refers to a heteroaryl radical as defined above and where the point of attachment of the heteroaryd radical to the rest of the molecule is through a carbon atom in the heteroaryl radical. A C-heteroaryd radical is optionally substituted as described above for heteroaryl radicals.

[0276] “Heteroarylalkyl” refers to a radical of the formula -Rc-heteroaryl, where Rcis an alkydene chain as defined above. If the heteroary 1 is a nitrogen-containing heteroaryd, the heteroaryl is optionally attached to the alkyl radical at the nitrogen atom. The alkylene chain of the heteroarylalkyl radical is optionally substituted as defined above for an alkylene chain. The heteroaryd part of the heteroarylalkyl radical is optionally substituted as defined above for a heteroaryd group.

[0277] The compounds, or their pharmaceutically acceptable salts may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as ( / )- or (< S)- or, as (D)- or (L)- for amino acids. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry', and unless specified otherwise, it is intended that the compounds include both E (or trans) and Z (cis) geometric isomers. Likewise, all possible isomers, as well as their racemic and optically pure forms, and all tautomeric forms are also intended to be included.

[0278] A “stereoisomer” refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable. It is therefore contemplated that various stereoisomers and mixtures thereof and includes “enantiomers,” which refers to two stereoisomers whose molecules are non-superimposable mirror images of one another.

[0279] A “tautomer” refers to a proton shift from one atom of a molecule to another atom of the same molecule. The compounds presented herein may exist as tautomers. Tautomers are compounds that are interconvertible by migration of a hydrogen atom, accompanied by a switch of a single bond and adjacent double bond. In solutions where tautomerization is possible, a chemical equilibrium of the tautomers will exist. The exact ratio of the tautomers depends on several factors, including temperature, solvent, and pH. Some examples of tautomeric pairs include:

[0280] “Optional” or “optionally” means that a subsequently described event or circumstance may or may not occur and that the description includes instances when the event or circumstance occurs and instances in which it does not.

[0281] “Pharmaceutically acceptable salt” includes both acid and base addition salts. A pharmaceutically acceptable salt of any one of the alkoxyphenyl-linked amine derivative compounds described herein is intended to encompass any and all pharmaceutically suitable salt forms. Preferred pharmaceutically acceptable salts of the compounds described herein are pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0282] “Pharmaceutically acceptable acid addition salt” refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, and the like. Also included are salts that are formed wi th organic acids such as aliphatic mono-and dicarbox lic acids, phenyl-substituted alkanoic acids, hydroxy7alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and. aromatic sulfonic acids, etc. and include, for example, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Exemplary salts thus include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, trifluoroacetates, propionates, caprylates, isobutyrates, oxalates,malonates, succinate suberates, sebacates, fumarates, maleates, mandelates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, malates, tartrates, methanesulfonates, and the like. Also contemplated are salts of amino acids, such as arginates, gluconates, and galacturonates (see, for example, Berge S. M. et al.. "Pharmaceutical Salts,” Journal of Pharmaceutical Science, 66: 1-19 (1997), which is hereby incorporated by reference in its entirety). Acid addition salts of basic compounds may be prepared by contacting the free base forms with a sufficient amount of the desired acid to produce the salt according to methods and techniques with which a skilled artisan is familiar.

[0283] “Pharmaceutically acceptable base addition salt” refers to those salts that retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Pharmaceutically acceptable base addition salts may be formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Salts derived from organic bases include, but are not limited to, salts of primary, secondary', and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, A, A-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline, A-methylglucamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine. A-ethylpiperidine, polyamine resins and the like. See Berge et al., supra.

[0284] As used herein, “treatment” or “treating,” or “palliating” or “ameliorating” are used interchangeably herein. These terms refer to an approach for obtaining beneficial or desired results including but not limited to therapeutic benefit and / or a prophylactic benefit. By “therapeutic benefit” is meant eradication, reduction, or amelioration of the underlying disorder being treated. Also, a therapeutic benefit is achieved with the eradication, reduction, or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the patient, notwithstanding that the patient may still be afflicted with the underlying disorder. For prophylactic benefit, the compositions may be administered to a patient at risk of developing a particular disease, or to a patient reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease may not have been made. Treatment and therapeutic benefit also include, e.g., in theinstance of cancer therapy or treatment of other disease or condition, remission in, reducing, ameliorating, or ablating the disease or condition, including remission, increasing quality of life, reducing the extent of resection, or number of surgeries required, extending progression free survival, or exhibiting other clinical or quality of life benefit to patent.

[0285] As used herein, the abbreviations for the natural L-enantiomeric amino acids are conventional and are as follows: alanine (A, Ala); arginine (R, Arg); asparagine (N, Asn); aspartic acid (D, Asp); cysteine (C, Cys); glutamic acid (E, Glu); glutamine (Q, Gin); glycine (G, Gly); histidine (H, His); isoleucine (I, He); leucine (L. Leu); lysine (K, Lys); methionine (M, Met): phenylalanine (F, Phe); proline (P, Pro); serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y, Tyr); valine (V, Vai). Typically, Xaa can indicate any amino acid. In some embodiments, X can be asparagine (N), glutamine (Q), histidine (H), lysine (K), or arginine (R).

[0286] Some embodiments of the disclosure contemplate D-amino acid residues of any standard or non-standard amino acid or analogue thereof. When an amino acid sequence is represented as a series of three-letter or one-letter amino acid abbreviations, the left-hand direction is the amino terminal direction and the right-hand direction is the carboxy terminal direction, in accordance with standard usage and convention.

[0287] The terms "‘peptide'’, “polypeptide”, “miniprotein”, and “protein” can be used interchangeably herein to refer to a polymer of amino acid residues. In various embodiments, “peptides”, “polypeptides”, and “proteins” can be chains of amino acids whose alpha carbons are linked through peptide bonds. The terminal amino acid at one end of the chain (e.g., amino terminal, or N-terminal) therefore can have a free amino group, while the terminal amino acid at the other end of the chain (e.g., carboxy terminal, or C-terminal) can have a free carboxyl group. As used herein, the term “amino terminus” (e.g., abbreviated N-tenninus) can refer to the free alpha, beta, or gamma-amino group on an amino acid at the amino terminal of a peptide or to the alpha, beta, or gamma-amino group (e.g., imino group when participating in a peptide bond) of an ammo acid at any other location within the peptide. Similarly, the term “carboxy terminus” can refer to the free carboxyl group on the carboxy terminus of a peptide or the carboxyl group of an amino acid at any other location within the peptide. Peptides also include essentially any poly amino acid including, but not limited to, peptide mimetics such as amino acids joined by an ether or thioether as opposed to an amide bond.

[0288] As used herein, the term “peptide construct” can refer to a molecule comprising one or more peptides of the present disclosure that can be conjugated to, linked to (including by complexation), or fused to one or more peptides or cargo molecules. In some cases, cargomolecules are active agents. The term '‘active agent" can refer to any molecule, e.g., any molecule that is capable of eliciting a biological effect and / or a physical effect (e.g., emission of radiation) which can allow the localization, detection, or visualization of the respective peptide construct. In various embodiments, the term “active agent” refers to a therapeutic and / or diagnostic agent.

[0289] As used herein, the term '‘peptide complex” can refer to one or more peptides of the present disclosure that are fused, linked, conjugated, or otherwise connected to form a complex.

[0290] As used herein, the terms “comprising” and “having” can be used interchangeably. For example, the terms “a peptide comprising an amino acid sequence of SEQ ID NO: 32” and “a peptide having an amino acid sequence of SEQ ID NO: 32” can be used interchangeably.

[0291] The term “engineered,” when applied to a polynucleotide, denotes that the polynucleotide has been removed from its natural genetic milieu and is thus free of other extraneous or unwanted coding sequences and is in a form suitable for use within genetically engineered protein production systems. Such engineered molecules are those that are separated from their natural environment and include cDNA and genomic clones (i.e., a prokaryotic or eukaryotic cell with a vector containing a fragment of DNA from a different organism).Engineered DNA molecules of the present invention are free of other genes with which they are ordinarily associated but can include naturally occurring or non-naturally occurring 5 ’and 3’ untranslated regions such as enhancers, promoters, and terminators.

[0292] An “engineered” polypeptide or protein is a polypeptide or protein that is found in a condition other than its native environment, such as apart from blood and animal tissue. In a preferred form, the engineered polypeptide is substantially free of other polypeptides, particularly other polypeptides of animal origin. It is preferred to provide the polypeptides in a highly purified form, e.g., greater than 90% pure, greater than 95% pure, more preferably greater than 98% pure or greater than 99% pure. When used in this context, the term “engineered” does not exclude the presence of the same polypeptide in alternative physical forms, such as dimers, heterodimers and multimers, heteromultimers, or alternatively glycosylated, carboxylated. modified, or derivatized forms.

[0293] An “engineered” peptide or protein is a polypeptide that is distinct from a naturally occurring polypeptide structure, sequence, or composition. Engineered peptides include non-naturally occurring, artificial, isolated, synthetic, designed, modified, or recombinantly expressed peptides.

[0294] Polypeptides of the disclosure include polypeptides that have been modified in any way, for example, to: (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation,(3) alter binding affinity for forming protein complexes, (4) alter binding affinities, (5) alter binding affinity at certain pH values, and (6) confer or modify other physicochemical or functional properties. For example, single or multiple amino acid substitutions (e.g., conservative amino acid substitutions) are made in the naturally occurring sequence (e g., in the portion of the polypeptide outside the domain(s) forming intermolecular contacts). A “conservative amino acid substitution” can refer to the substitution in a polypeptide of an amino acid with a functionally similar amino acid. The following six groups each contain amino acids that can be conservative substitutions for one another: i) Alanine (A), Serine (S), and Threonine (T); ii) Aspartic acid (D) and Glutamic acid (E): iii) Asparagine (N) and Glutamine (Q); iv) Arginine (R) and Lysine (K); v) Isoleucine (I), Leucine (L), Methionine (M), and Valine (V); vi) Phenylalanine (F), Tyrosine (Y), and Tryptophan (W). In some embodiments, a conserved amino acid substitution can comprise a non-natural amino acid. For example, substitution of an amino acid for a non-natural derivative of the same amino acid can be a conserved substitution.

[0295] The terms “polypeptide fragment” and “truncated polypeptide” as used herein can refer to a polypeptide that has an amino-terminal and / or carboxy -terminal deletion as compared to a corresponding full-length peptide or protein. In various embodiments, fragments are at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50. at least 100, at least 150. at least 200, at least 250. at least 300, at least 350. at least 400, at least 450, at least 500, at least 600, at least 700, at least 800, at least 900 or at least 1000 amino acids in length. In various embodiments, fragments can also be, e.g., at most 1000, at most 900, at most 800, at most 700, at most 600, at most 500, at most 450, at most 400, at most 350, at most 300, at most 250. at most 200, at most 150, at most 100. at most 50. at most 45. at most 40. at most 35, at most 30, at most 25, at most 20, at most 15, at most 10, or at most 5 amino acids in length. A fragment can further comprise, at either or both of its ends, one or more additional amino acids, for example, a sequence of amino acids from a different naturally occurring protein (e.g., an Fc or leucine zipper domain) or an artificial amino acid sequence (e.g., an artificial linker sequence).

[0296] As used herein, the terms “peptide” or “polypeptide” in conjunction with “variant”, “mutant”, or “enriched mutant”, or “permuted enriched mutant” can refer to a peptide or polypeptide that can comprise an amino acid sequence wherein one or more amino acid residues are inserted into, deleted from and / or substituted into the amino acid sequence relative to another polypeptide sequence. In various embodiments, the number of amino acid residues to be inserted, deleted, or substituted is at least 1, at least 2, at least 3, at least 4, at least 5, at least 10, at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200,at least 225, at least 250, at least 275, at least 300, at least 350, at least 400, at least 450 or at least 500 amino acids in length. Variants of the present disclosure include peptide conjugates or fusion molecules (e.g., peptide constructs or peptide complexes).

[0297] A “derivative” of a peptide or polypeptide can be a peptide or polypeptide that can have been chemically modified, e.g., conjugation to another chemical moietv such as, for example, polyethylene glycol, albumin (e.g., human serum albumin), phosphorylation, and glycosylation.

[0298] The term “% sequence identity ” can be used interchangeably herein with the term “% identity ” and can refer to the level of amino acid sequence identity between two or more peptide sequences or the level of nucleotide sequence identity between two or more nucleotide sequences, when aligned using a sequence alignment program. For example, as used herein, 80% identity7means the same thing as 80% sequence identity determined by a defined algorithm and means that a given sequence is at least 80% identical to another length of another sequence. In various embodiments, the % identity is selected from, e.g., at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99% or more up to 100% sequence identity7to a given sequence. In various embodiments, the % identity is in the range of, e.g., about 60% to about 70%, about 70% to about 80%, about 80% to about 85%. about 85% to about 90%, about 90% to about 95%. or about 95% to about 99%.

[0299] The terms “% sequence homology” or “percent sequence homology ” or “percent sequence identity ” can be used interchangeably herein with the terms “% homology,” “% sequence identity,” or “% identity” and can refer to the level of amino acid sequence homology between two or more peptide sequences or the level of nucleotide sequence homology between two or more nucleotide sequences, when aligned using a sequence alignment program. For example, as used herein, 80% homology7means the same thing as 80% sequence homology7determined by a defined algorithm, and accordingly a homologue of a given sequence has greater than 80% sequence homology over a length of the given sequence. In various embodiments, the % homology is selected from, e.g., at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% or more up to 100% sequence homology to a given sequence. In various embodiments, the % homology is in the range of, e.g.. about 60% to about 70%, about 70% to about 80%. about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, or about 95% to about 99%.

[0300] A protein or polypeptide can be “substantially pure,” “substantially homogeneous”, or “substantially purified” when at least about 60% to 75% of a sample exhibits a single species ofpolypeptide. The polypeptide or protein can be monomeric or multimeric. A substantially pure polypeptide or protein can typically comprise about 50%, 60%, 70%, 80% or 90% W / W of a protein sample, more usually about 95%, and e.g., will be over 98% or 99% pure. Protein purity or homogeneity’ can be indicated by a number of means well known in the art, such as polyacrylamide gel electrophoresis of a protein sample, followed by visualizing a single polypeptide band upon staining the gel with a stain well known in the art. For certain purposes, higher resolution is provided by using high-pressure liquid chromatography (e.g., HPLC) or other high-resolution analytical techniques (e.g., LC-mass spectrometry ).

[0301] As used herein, the term “pharmaceutical composition"’ can generally refer to a composition suitable for pharmaceutical use in a subject such as an animal (e.g., human or mouse). A pharmaceutical composition can comprise a pharmacologically effective amount of an active agent and a pharmaceutically acceptable carrier. The term “pharmacologically effective amount” can refer to that amount of an agent effective to produce the intended biological or pharmacological result.

[0302] As used herein, the term “pharmaceutically acceptable carrier” can refer to any of the standard pharmaceutical carriers, vehicles, buffers, and excipients, such as a phosphate buffered saline solution, or a buffered saline solution, 5% aqueous solution of dextrose, and emulsions, such as an oil / water or water / oil emulsion, and various types of wetting agents and / or adjuvants. Suitable pharmaceutical carriers and formulations are described in Remington's Pharmaceutical Sciences, 21st Ed. 2005, Mack Publishing Co, Easton. A “pharmaceutically acceptable salt” can be a salt that can be formulated into a compound for pharmaceutical use including, e.g., metal salts (sodium, potassium, magnesium, calcium, etc.) and salts of ammonia or organic amines.

[0303] As used herein, the terms “treat”, “treating” and “treatment” can refer to a method of alleviating or abrogating a biological disorder and / or at least one of its attendant symptoms. As used herein, to “alleviate” a disease, disorder, or condition, for example, means reducing the severity’ and / or occurrence frequency of the symptoms of the disease, disorder, or condition. Further, references herein to “treatment” can include references to curative, palliative, and prophylactic or diagnostic treatment.

[0304] Generally, a cell of the present disclosure can be a eukaryotic cell or a prokaryotic cell. A cell can be an epithelial cell. A cell can be a microorganism, bacterial, yeast, fungal or algae cell. A cell can be an animal cell or a plant cell. An animal cell can include a cell from a marine invertebrate, fish, insects, amphibian, reptile, or mammal. A mammalian cell can be obtained from a primate, ape, equine, bovine, porcine, canine, feline, or rodent. A mammal can be a primate, ape, dog, cat, rabbit, ferret, or the like. A rodent can be a mouse, rat, hamster, gerbil,hamster, chinchilla, or guinea pig. A bird cell can be from a canary, parakeet, or parrots. A reptile cell can be from a turtles, lizard, or snake. A fish cell can be from a tropical fish. For example, the fish cell can be from a zebrafish (e.g., Danino rerio). A worm cell can be from a nematode (e.g., C. elegcms). An amphibian cell can be from a frog. An arthropod cell can be from a tarantula or hermit crab.

[0305] A mammalian cell can also include cells obtained from a primate (e.g., a human or a non-human primate). A mammalian cell can include a blood cell, a stem cell, an epithelial cell, connective tissue cell, hormone secreting cell, a nerve cell, a skeletal muscle cell, or an immune system cell.

[0306] As used herein, the term '‘vector,” generally refers to a DNA molecule capable of replication in a host cell and / or to which another DNA segment can be operatively linked so as to bring about replication of the attached segment. A plasmid is an exemplary vector.

[0307] As used herein, the term “subject,” generally refers to a human or to another animal. A subject can be of any age, for example, a subject can be prenatal, newborn, an infant, a toddler, a child, a pre-adolescent, an adolescent, an adult, or an elderly individual.

[0308] Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are in relation to the other endpoint, and independently of the other endpoint. The term “about” as used herein refers to a range that is 15% plus or minus from a stated numerical value within the context of the particular usage. For example, about 10 can include a range from 8.5 to 11.5.Nerve Homing Agents

[0309] Over 300 million surgeries are performed worldwide each year. Despite many recent advances in the treatment of cancer and other diseases, surgery remains the most effective treatment option for a number of diseases and injuries. The ultimate goal of surgery is to remove or repair tissues while minimizing comorbidities by preserving vital structures such as nerves and blood vessels. Recent technological advances including minimally invasive robot assisted laparoscopic surgery have improved outcomes and made it possible to perform difficult procedures robustly with minimal risk. Furthermore, preoperative three-dimensional imaging technologies such as magnetic resonance imaging (MRI) and computed tomography (CT) have vastly improved diagnostic accuracy, staging, and preoperative planning.

[0310] While advances have been made, identifying vital structures for preservation (e.g., nerves) or tissue for complete resection (e.g., tumors) during surgical procedures remains difficult. Nen e identification and sparing can be difficult intraoperatively due to variations in patient anatomy and often little ability for direct nerve visualization in the surgical field.Currently, intraoperative nerve detection is performed through a combination of naked eye visualization, palpation, and electromyographic monitoring. Several imaging modalities have been utilized in clinical studies for nerve detection including ultrasound, optical coherence tomography, and confocal endomicroscopy. However, these lack specificity, resolution, and wide-field imaging functionality, making it difficult to identify nerve tissues in real time. As a result, nerve damage continues to plague surgical outcomes. Iatrogenic nerve injury affects up to 63 million patients worldwide annually, causing acute and chronic pain as well as impairment or loss of motor and sensory function. Radical prostatectomy (RP), a surgical procedure involving removal of the entire prostate as a prostate cancer cure, is particularly plagued by nerve damage. Furthermore, while minimally invasive methods, such as robotic assisted RP, can achieve equivalent cancer control to open RP while resulting in decreased blood loss, lower transfusion rate, and faster convalescence, these advances provide no benefit in nerve-sparing outcomes and in fact, remove the ability to directly palpate the tissue.

[0311] An imaging modality capable of wide field, real time identification of nerve tissues intraoperatively would greatly benefit surgeons in nerve preservation and reduce rates of iatrogenic nerve injury, improving quality of life for patients post-surgery.

[0312] Fluorescence imaging in the NIR region (650-900 nm) is advantageous as endogenous tissue chromophore absorbance, scattering and autofluorescence are all at local minima, creating a black background upon which tissue-specific contrast can be added. The minimal photon scatter and absorbance in the NIR region also facilitates photon penetration for fluorescence imaging up to centimeters deep in tissue as compared to a few' hundred microns using visible light. In the context of image-guided surgery, NIR fluorescence enables real-time, non-contact imaging where the addition of fluorescent contrast and NIR light does not alter the look of the surgical field and contrast agent detection does not require ionizing radiation.Tumor Homing Agents

[0313] The present disclosure provides methods for administering compounds that selectively bind to certain types of cancer or cancer cells and tissues (e g., tumor homing agents). In some embodiments, the cancer is glioma, astrocytoma, medulloblastoma, choroids plexus carcinoma, ependymoma, neuroblastoma, vestibular schwannoma, carcinoma, meningioma, head and neck cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, breast cancer, intestinalcancer, pancreatic cancer, liver cancer, kidney cancer, sarcoma, osteosarcoma, rhabdomyosarcoma, Ewing’s sarcoma, gastrointestinal stromal tumors, melanoma, ovarian cancer, centical cancer, lymphoma, thyroid cancer, anal cancer, colorectal cancer, endometrial cancer, laryngeal cancer, multiple myeloma, prostate cancer, retinoblastoma, gastric cancer, testicular cancer. Wilm’s tumor, or any combination thereof. In some embodiments, the cancer is head and neck cancer, a brain cancer, a brain tumor, breast cancer, melanoma, sarcoma, basal cell carcinoma, squamous cell carcinoma, lung cancer, colorectal cancer, prostate cancer, bladder cancer, or any combination thereof. In some embodiments, the head and neck cancer is head and neck squamous cell carcinoma. In some embodiments, the breast cancer is invasive ductal carcinoma, ductal carcinoma in situ breast cancer, invasive lobular carcinoma, lobular carcinoma in situ breast cancer, or any combination thereof.

[0314] As yet another example, the present disclosure provides a method for administering compounds that selectively bind to a cavernoma (also referred to as cavernous angiomas, cavernous hemangiomas, or cerebral cavernous malformation (CCM)) cells and tissues, arteriovenous malformation (also referred to as arteriovenous angiomas, arteriovenous hemangiomas, or cerebral arteriovenous malformation (CAM)) cells and tissues, an aneury sm (e.g., including abdominal aortic, thoracic aortic, and cerebral aneury sm) cells and tissues, or a spinal dural arteriovenous fistula cells and tissues. In various aspects, these compounds can comprise a peptide portion and a detectable agent conjugated together.

[0315] In various aspects of the compounds used in the present disclosure, the peptide portions of the compounds described herein have certain features in common with the native chlorotoxin (CTX) peptide (SEQ ID NO: 1). The native chlorotoxin peptide was originally isolated from the scorpion Leiurus quinquestriatus. Chlorotoxin is a 36 amino acid peptide that selectively binds to or accumulates in cancerous cells. In certain aspects, peptides used in the present disclosure are conjugated to moieties, such as detectable labels (e.g., dyes or radiolabels) that are detected (e.g., visualized) in a subject. In some aspects, the peptides including chlorotoxin and / or chlorotoxin variants are conjugated to detectable labels to enable tracking of the bio-distribution of a conjugated peptide. The fluorescent moiety can be covalently coupled to the peptide and / or peptide variants to allow for the visualization of the conjugate by fluorescence imaging, either directly or through a linker as described herein and known to one of ordinary skill in the art. Linker moieties can include cleavable (e.g., pH sensitive or enzyme-labile linkers) or stable linkers.

[0316] In some aspects, the fluorescent label used has emission characteristics that are desired for a particular application. Fluorophores can be conjugated or fused to another moiety asdescribed herein and be used to home, target, migrate to, be retained by, accumulate in, and / or bind to, or be directed to specific organs, substructures within organs, tissues, nerves, targets or cells and used in conjunction with the compounds and methods herein. Exemplary organs and organ substructures include the brain and other organs and organ structures such as brain, heart, lung, kidney, liver. CNS (e.g.. spine) or pancreas or in the extremities (e.g., legs. neck, and arms). The fluorophore emission can comprise an infrared, near infrared, blue or ultraviolet emission.

[0317] In some aspects, the peak spectral emission of the nene homing agent may be blue shifted relative to the tumor homing agent, or the tumor homing agent may be blue shifted relative to the nerve homing agent. In some aspects, the minimum spectral gap in emission wavelength of the nerve homing agent relative to the tumor homing agent is at least about 0 nm, at least about 10 nm, at least about 20 nm, at least about 30 nm, at least about 40 nm, at least about 50 nm. at least about 60 nm. at least about 70 nm. at least about 80 nm, at least about 90 nm, at least about 100 nm, at least about 110 nm, at least about 120 nm, at least about 130 nm, at least about 140 nm, at least about 150 nm, at least about 160 nm, at least about 170 nm, at least about 180 nm, at least about 190 nm, at least about 200 nm, at least about 210 nm, at least about 220 nm, at least about 230 nm, at least about 240 nm, at least about 250 nm, at least about 260 nm, at least about 270 nm. at least about 280 nm, at least about 290 nm, at least about 300 nm. at least about 310 nm, at least about 320 nm, at least about 330 nm, at least about 340 nm, at least about 350 nm, at least about 360 nm, at least about 370 nm, at least about 380 nm, at least about 390 nm, at least about 400 nm, at least about 410 nm, at least about 420 nm, at least about 430 nm, at least about 440 nm, at least about 450 nm, at least about 460 nm, at least about 470 nm. at least about 480 nm, at least about 490 nm, at least about 500 nm, at least about 510 nm, at least about 520 nm, at least about 530 nm, at least about 540 nm, at least about 550 nm, at least about 560 nm, at least about 570 nm, at least about 580 nm, at least about 590 nm, at least about 600 nm, at least about 610 nm, at least about 620 nm, at least about 630 nm, at least about 640 nm, at least about 650 nm, at least about 660 nm. at least about 670 nm, at least about 680 nm, at least about 690 nm, at least about 700 nm, at least about 710 nm, at least about 720 nm, at least about 730 nm, at least about 740 nm, at least about 750 nm, at least about 760 nm, at least about 770 nm, at least about 780 nm, at least about 790 nm, at least about 800 nm, at least about 810 nm, at least about 820 nm, at least about 830 nm. at least about 840 nm, at least about 850 nm, at least about 860 nm, at least about 870 nm, at least about 880 nm, at least about 890 nm, or any wavelength gap within any of these forgoing ranges. Between any two of these values withinthis minimum spectral gap in emission wavelength, could be + / - 10 nm, + / - 3 nm, + / - 2 nm, or + / - 1 nm, or + / - 10%, + / - 5%, + / - 1% in the context of the compounds and methods herein.

[0318] In some aspects, the peak excitation wavelength of the nerve homing agent may be blue shifted relative to the tumor homing agent, or the tumor homing agent may be blue shifted relative to the nerve homing agent. In some aspects, the minimum spectral gap in excitation wavelength of the nerve homing agent and tumor homing agent may be separated by at least about 0 nm, at least about 10 nm, at least about 20 nm, at least about 30 nm, at least about 40 nm, at least about 50 nm, at least about 60 nm, at least about 70 nm, at least about 80 nm, at least about 90 nm. at least about 100 nm, at least about 110 nm, at least about 120 nm. at least about 130 nm, at least about 140 nm, at least about 150 nm, at least about 160 nm, at least about 170 nm, at least about 180 nm, at least about 190 nm, at least about 200 nm, at least about 210 nm, at least about 220 nm, at least about 230 nm, at least about 240 nm, at least about 250 nm, at least about 260 nm, at least about 270 nm, at least about 280 nm. at least about 290 nm, at least about 300 nm, at least about 310 nm, at least about 320 nm, at least about 330 nm, at least about 340 nm, at least about 350 nm, at least about 360 nm, at least about 370 nm, at least about 380 nm, at least about 390 nm, at least about 400 nm, at least about 410 nm, at least about 420 nm, at least about 430 nm, at least about 440 nm, at least about 450 nm, at least about 460 nm, at least about 470 nm, at least about 480 nm, at least about 490 nm. at least about 500 nm, at least about 510 nm, at least about 520 nm, at least about 530 nm, at least about 540 nm, at least about 550 nm, at least about 560 nm, at least about 570 nm, at least about 580 nm, at least about 590 nm, at least about 600 nm, at least about 610 nm, at least about 620 nm, at least about 630 nm, at least about 640 nm, at least about 650 nm, at least about 660 nm, at least about 670 nm, at least about 680 nm, at least about 690 nm, at least about 700 nm, at least about 710 nm, at least about 720 nm, at least about 730 nm, at least about 740 nm, at least about 750 nm, at least about 760 nm, at least about 770 nm, at least about 780 nm, at least about 790 nm, at least about 800 nm, at least about 810 nm, at least about 820 nm, at least about 830 nm, at least about 840 nm, at least about 850 nm, at least about 860 nm. at least about 870 nm, at least about 880 nm, at least about 890 nm. or any wavelength gap within any of these forgoing ranges. Between any two of these values within this minimum spectral gap in excitation wavelength, could be + / - 10 nm, + / - 3 nm, + / - 2 nm, or + / - 1 nm, or + / - 10%, + / - 5%, + / - 1% in the context of the compounds and methods herein.

[0319] In some aspects, the excitation wavelength spectrum of the nerve homing agent overlaps the emission wavelength spectrum of the tumor homing agent by at least about 0%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at leastabout 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 21%. at least about 22%, at least about 23%, at least about 24%, at least about 25%. at least about 26%. at least about 27%, at least about 28%, at least about 29%, at least about 30%, at least about 31%, at least about 32%, at least about 33%, at least about 34%, at least about 35%, at least about 36%, at least about 37%, at least about 38%, at least about 39%, at least about 40%, at least about 41%, at least about 42%, at least about 43%, at least about 44%. at least about 45%. at least about 46%, at least about 47%, at least about 48%, at least about 49%, at least about 50%, at least about 51%, at least about 52%, at least about 53%, at least about 54%, at least about 55%, at least about 56%, at least about 57%, at least about 58%, at least about 59%, at least about 60%, at least about 61%, at least about 62%, at least about 63%, at least about 64%. at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%. at least about 84%, at least about 85%, at least about 86%, at least about 87%. at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, at least about 100%, or a range between any two of these values. Betw een any two of these values within this overlap range, it is recognized that the values could be + / - 0.1%, + / - 0.2%, + / - 0.3%, + / - 0.4%. + / - 0.5%, + / - 0.6%, + / - 0.7%, + / - 0.8%, + / - 0.9% relative to one or more of the integer value above, in the context of the compounds and methods herein

[0320] In some aspects, the excitation spectrum of the nerve homing agent does not overlap the spectrum emission wavelength of the tumor homing agent. In some aspects, the excitation spectrum of the nerve homing agent completely overlaps the spectrum emission wavelength of the tumor homing agent. In some aspects, the excitation spectrum of the nen e homing agent completely overlaps the spectrum excitation wavelength of the tumor homing agent. It is understood that a minimum difference in the emission spectrum could be as low as 0 nm (i.e., complete spectral overlap between the excitation spectra of the tumor homing agent and nerve homing agent), as there potentially exist hyperspectral systems that can distinguish multiple excitation profdes from each other even when overlapping. Additionally, in the context of surgery, surgeons can additionally distinguish overlap based on anatomical differences of tumorand nerve tissues, cells and structures to aid in distinguishing and identifying the nerve specific and tumor specific signals in addition to using spectral differences which through use of the methods described herein can improve surgical outcomes in cancer surgeries and reduce comorbidities for patients.

[0321] In some aspects, the emission spectrum of the nerve homing agent overlaps the spectrum excitation wavelength of the tumor homing agent by at least about 0%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%. at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 21%, at least about 22%, at least about 23%, at least about 24%, at least about 25%, at least about 26%, at least about 27%, at least about 28%, at least about 29%, at least about 30%, at least about 31%. at least about 32%, at least about 33%, at least about 34%, at least about 35%, at least about 36%, at least about 37%, at least about 38%, at least about 39%, at least about 40%, at least about 41%, at least about 42%, at least about 43%, at least about 44%, at least about 45%, at least about 46%, at least about 47%, at least about 48%, at least about 49%, at least about 50%. at least about 51%, at least about 52%, at least about 53%, at least about 54%, at least about 55%, at least about 56%. at least about 57%. at least about 58%, at least about 59%, at least about 60%, at least about 61%, at least about 62%, at least about 63%, at least about 64%, at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%. at least about 76%. at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%. at least about 96%, at least about 97%, at least about 98%. at least about 99%, at least about 99.5%, at least about 100%. or a range between any two of these values. Between any two of these values within this overlap range, it is recognized that the values could be + / - 0.1%, + / - 0.2%, + / - 0.3%, + / - 0.4%, + / - 0.5%, + / - 0.6%, + / - 0.7%. + / - 0.8%, + / - 0.9% relative to one or more of the integer value above, in the context of the compounds and methods herein.

[0322] In some aspects, the emission spectrum of the nerve homing agent does not overlap the spectrum excitation wavelength of the tumor homing agent. In some aspects, the emission spectrum of the nerve homing agent completely overlaps the spectrum emission wavelength ofthe tumor homing agent. It is understood that a minimum difference in the emission spectrum could be as low as 0 nm (i.e., complete spectral overlap between the emission spectra of the tumor homing agent and nerve homing agent), as there potentially exist hyperspectral systems that can distinguish multiple emission profiles from each other even when overlapping.Additionally, in the context of surgery, surgeons can additionally distinguish overlap based on anatomical differences of tumor and nerve tissues, cells and structures to aid in distinguishing and identifying the nerve specific and tumor specific signals in addition to using spectral differences which through use of the methods described herein can improve surgical outcomes in cancer surgeries and reduce comorbidities for patients.

[0323] It is understood that that different agents may have different spectral properties having smaller or larger stokes shift, which would allow a discernable difference in emission wavelength even if excited at the same wavelength.

[0324] In some embodiments, the compounds and methods herein are used as imaging agents to detect their fluorophore emissions. The fluorophores emissions can comprise a near infrared (NIR) emission. In various aspects, In some embodiments, compounds and methods herein used as imaging agents to detect tumor or nerve tissue or cells individually comprise fluorophores that have an absorption wavelength of between about 650 and 800 nm, or 600 nm and about 800 nm, or between about 700 nm to about 800 nm, about 800 nm to about 950 nm, about 800 nm to about 880 nm, about 775 nm to about 795 nm, or about 785 nm. In another aspect, the imaging agents to detect tumor or nerve tissue or cells individually comprise fluorophores having an absorption wavelength 650 nm, 675 nm, 700 nm, 725 nm, 750 nm, 775 nm, 780 nm, 785 nm, 790 nm, 795 nm. 800 nm, 805 nm, 810 nm, 815 nm, 820 nm, 825 nm, 830 nm, 835 nm, 840 nm.850 nm, 855 nm, 860 nm, 865 nm, 870 nm, 875 nm, 880 nm, 885 nm, 890 nm, 895 nm, or 900 nm. In some embodiments, each fluorophore has wavelengths in the ranges disclosed herein, including 600-610 nm, 610-620 nm, 620-630 nm, 630-640 nm, 640-650 nm, 650-660 nm, 660-670 nm, 670-680 nm, 680-690 nm, 690-700 nm, 700-710 nm, 710-720 nm. 720-730 nm, 730- 740 nm, 740-750 nm, 750-760 nm, 760-770 nm, 770-780 nm, 780-790 nm. 790-800 nm. 800- 810 nm, 810-820 nm, 820-830 nm, 830-840 nm, 840-850 nm, 850-860 nm, 600-650 nm, 625- 675 nm, 650-700 nm, 700-750 nm, 750-800 nm, 800-850 nm, or 850-900 nm, or any wavelength within any of these foregoing ranges. With respect to the above absorption spectra, fluorophores that have an absorption wavelength, could be + / - 10 nm, + / - 3 nm, + / - 2 nm, or + / - 1 nm. or + / -10%, + / - 5%, + / - 1% in the context of the compounds and methods herein.

[0325] The fluorophores emissions can comprise an ultraviolet emission. The ultraviolet emissions can have a wavelength from 10 nm to 400 nm, and up to 450 nm or 460 nm into the -1 liblue light spectrum, including fluorophores with absorption wavelengths in the ranges disclosed herein, including 10-20 nm, 20-30 nm, 30-40 nm, 40-50 nm, 50-60 nm, 60-70 nm, 70-80 nm, 80-90 nm, 90-100 nm, 100-110 nm, 110-120 nm, 120-130 nm, 130-140 nm, 140-150 nm, 150-160 nm, 160-170 nm, 170-180 nm, 180-190 nm, 190-200 nm, 200-210 nm. 210-220 nm, 220-230 nm, 230-240 nm, 240-250 nm, 250-260 nm, 260-270 nm, 270-280 nm. 280-290 nm. 290-300 nm, 300-310 nm, 310-320 nm, 320-330 nm, 330-340 nm, 340-350 nm, 350-360 nm, 360-370 nm, 370-380 nm, 380-390 nm, 390-400 nm, 400-410 nm, 410-420 nm, 420-430 nm, 430-440 nm, 440-450 nm, 450-460 nm, 300-350 nm, 325-375 nm, 350-400 nm, 400-450 nm, a wavelength in the range of 340 nm to 400 nm. 360 to 420 nm, 380 nm to 440 nm, 400 nm to 450 nm, 400 nm to 460 nm or any wavelength within any of these foregoing ranges. With respect to the above absorption spectra, fluorophores that have an absorption wavelength, could be + / - 10 nm, + / - 3 nm, + / - 2 nm, or + / - 1 nm, or + / - 10%, + / - 5%, + / - 1% in the context of the compounds and methods herein.

[0326] In some embodiments, compounds and methods herein are used as imaging agents to detect their fluorophores have an absorption wavelength of about 200 nm to about 1,000 nm. In some embodiments, compounds and methods herein are used as imaging agents to detect their fluorophores have an absorption wavelength of about 200 nm to about 250 nm. about 200 nm to about 300 nm, about 200 nm to about 350 nm, about 200 nm to about 400 nm, about 200 nm to about 450 nm, about 200 nm to about 500 nm, about 200 nm to about 600 nm, about 200 nm to about 700 nm, about 200 nm to about 800 nm, about 200 nm to about 900 nm, about 200 nm to about 1,000 nm, about 250 nm to about 300 nm, about 250 nm to about 350 nm, about 250 nm to about 400 nm. about 250 nm to about 450 nm, about 250 nm to about 500 nm. about 250 nm to about 600 nm, about 250 nm to about 700 nm, about 250 nm to about 800 nm, about 250 nm to about 900 nm, about 250 nm to about 1,000 nm, about 300 nm to about 350 nm, about 300 nm to about 400 nm, about 300 nm to about 450 nm, about 300 nm to about 500 nm, about 300 nm to about 600 nm, about 300 nm to about 700 nm, about 300 nm to about 800 nm, about 300 nm to about 900 nm. about 300 nm to about 1,000 nm, about 350 nm to about 400 nm. about 350 nm to about 450 nm, about 350 nm to about 500 nm, about 350 nm to about 600 nm, about 350 nm to about 700 nm, about 350 nm to about 800 nm, about 350 nm to about 900 nm, about 350 nm to about 1,000 nm, about 400 nm to about 450 nm, about 400 nm to about 500 nm, about 400 nm to about 600 nm, about 400 nm to about 700 nm. about 400 nm to about 800 nm, about 400 nm to about 900 nm, about 400 nm to about 1,000 nm, about 450 nm to about 500 nm, about 450 nm to about 600 nm, about 450 nm to about 700 nm, about 450 nm to about 800 nm, about 450 nm to about 900 nm, about 450 nm to about 1,000 nm, about 500 nm to about600 nm, about 500 nm to about 700 nm, about 500 nm to about 800 nm, about 500 nm to about 900 nm, about 500 nm to about 1,000 nm, about 600 nm to about 700 nm, about 600 nm to about 800 nm, about 600 nm to about 900 nm, about 600 nm to about 1,000 nm, about 700 nm to about 800 nm, about 700 nm to about 900 nm, about 700 nm to about 1,000 nm, about 800 nm to about 900 nm. about 800 nm to about 1,000 nm, or about 900 nm to about 1.000 nm. In some embodiments, compounds and methods herein are used as imaging agents to detect their fluorophores have an absorption wavelength of about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, about 500 nm, about 600 nm, about 700 nm, about 800 nm, about 900 nm, or about 1.000 nm. In some embodiments, compounds and methods herein are used as imaging agents to detect their fluorophores have an absorption wavelength of at least about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, about 500 nm, about 600 nm, about 700 nm, about 800 nm, or about 900 nm or any wavelength within any of these foregoing ranges. In some embodiments, compounds and methods herein are used as imaging agents to detect their fluorophores have an absorption wavelength of at most about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, about 500 nm, about 600 nm, about 700 nm, about 800 nm, about 900 nm, or about 1,000 nm or any wavelength within any of these foregoing ranges. With respect to the above absorption spectra, fluorophores that have an absorption wavelength, could be + / - 150 nm. + / - 100 nm, + / - 90 nm, + / - 80 nm, + / -70 nm, + / - 60 nm, + / - 50 nm, + / - 40 nm, + / - 30 nm, + / - 20 nm, + / - 10 nm, + / - 3 nm, + / - 2 nm, or + / - 1 nm, or + / - 50%, + / - 25%, + / - 10%, + / - 5%, + / - 1% in the context of the compounds and methods herein.

[0327] In some embodiments, compounds and methods herein are used as imaging agents to detect their fluorophores have an absorption wavelength of about 1,000 nm to about 4,000 nm. In some embodiments, compounds and methods herein are used as imaging agents to detect their fluorophores have an absorption wavelength of about 1,000 nm to about 1,250 nm, about 1,000 nm to about 1,500 nm, about 1,000 nm to about 1.750 nm, about 1,000 nm to about 2,000 nm, about 1,000 nm to about 2.250 nm, about 1.000 nm to about 2,500 nm, about 1,000 nm to about 2,750 nm, about 1,000 nm to about 3,000 nm, about 1,000 nm to about 3,250 nm, about 1,000 nm to about 3,500 nm, about 1,000 nm to about 4,000 nm, about 1,250 nm to about 1,500 nm, about 1,250 nm to about 1.750 nm, about 1,250 nm to about 2,000 nm, about 1,250 nm to about 2,250 nm, about 1.250 nm to about 2,500 nm, about 1,250 nm to about 2.750 nm, about 1,250 nm to about 3,000 nm, about 1,250 nm to about 3,250 nm, about 1,250 nm to about 3,500 nm, about 1,250 nm to about 4,000 nm, about 1,500 nm to about 1,750 nm, about 1,500 nm to about 2,000 nm, about 1,500 nm to about 2,250 nm, about 1,500 nm to about 2,500 nm, about 1,500nm to about 2,750 nm, about 1,500 nm to about 3,000 nm, about 1,500 nm to about 3,250 nm, about 1,500 nm to about 3,500 nm, about 1,500 nm to about 4,000 nm, about 1,750 nm to about 2,000 nm, about 1,750 nm to about 2,250 nm, about 1,750 nm to about 2,500 nm, about 1,750 nm to about 2,750 nm, about 1,750 nm to about 3.000 nm, about 1,750 nm to about 3,250 nm, about 1,750 nm to about 3.500 nm, about 1.750 nm to about 4,000 nm, about 2,000 nm to about 2.250 nm, about 2,000 nm to about 2,500 nm, about 2,000 nm to about 2,750 nm, about 2,000 nm to about 3,000 nm, about 2,000 nm to about 3,250 nm, about 2,000 nm to about 3,500 nm, about 2,000 nm to about 4.000 nm, about 2,250 nm to about 2,500 nm, about 2,250 nm to about 2,750 nm, about 2.250 nm to about 3,000 nm, about 2,250 nm to about 3.250 nm, about 2,250 nm to about 3,500 nm, about 2,250 nm to about 4,000 nm, about 2,500 nm to about 2,750 nm, about 2,500 nm to about 3,000 nm, about 2,500 nm to about 3,250 nm, about 2,500 nm to about 3,500 nm, about 2,500 nm to about 4,000 nm, about 2,750 nm to about 3,000 nm, about 2,750 nm to about 3,250 nm, about 2,750 nm to about 3.500 nm, about 2,750 nm to about 4,000 nm, about 3,000 nm to about 3,250 nm, about 3,000 nm to about 3,500 nm, about 3,000 nm to about 4,000 nm, about 3,250 nm to about 3,500 nm, about 3,250 nm to about 4,000 nm, or about 3,500 nm to about 4,000 nm or any wavelength within any of these foregoing ranges. In some embodiments, compounds and methods herein are used as imaging agents to detect their fluorophores have an absorption wavelength of about 1.000 nm, about 1,250 nm. about 1,500 nm, about 1,750 nm, about 2,000 nm, about 2,250 nm, about 2,500 nm, about 2,750 nm, about 3,000 nm, about 3,250 nm, about 3,500 nm, or about 4,000 nm. In some embodiments, compounds and methods herein are used as imaging agents to detect their fluorophores have an absorption wavelength of at least about 1,000 nm. about 1,250 nm, about 1,500 nm, about 1,750 nm, about 2,000 nm, about 2,250 nm, about 2,500 nm, about 2,750 nm, about 3,000 nm, about 3.250 nm, or about 3,500 nm. In some embodiments, compounds and methods herein are used as imaging agents to detect their fluorophores have an absorption wavelength of at most about 1.250 nm, about 1,500 nm, about 1,750 nm, about 2,000 nm, about 2.250 nm, about 2,500 nm, about 2,750 nm, about 3.000 nm, about 3.250 nm. about 3,500 nm, or about 4,000 nm or any wavelength within any of these foregoing ranges. With respect to the above absorption spectra, fluorophores that have an absorption wavelength, could be + / - 150 nm, + / - 100 nm, + / - 90 nm, + / - 80 nm, + / - 70 nm, + / - 60 nm, + / - 50 nm, + / - 40 nm, + / - 30 nm, + / - 20 nm, + / - 10 nm, + / - 3 nm, + / - 2 nm, or + / - 1 nm, or + / - 50%, + / - 25%, + / - 10%. + / - 5%, + / - 1% in the context of the compounds and methods herein.

[0328] In some embodiments, the dyes disclosed herein have a peak absorbance of about 200 nm, about 210 nm, about 220 nm, about 230 nm, about 240 nm, about 250 nm, about 260 nm,about 270 nm, about 280 nm, about 290 nm, about 300 nm, about 310 nm, about 320 nm, about 330 nm, about 340 nm, about 350 nm, about 360 nm, about 370 nm, about 380 nm, about 390 nm, about 400 nm, about 410 nm, about 420 nm, about 430 nm, about 440 nm, about 450 nm, about 460 nm, about 470 nm, about 480 nm, about 490 nm, about 500 nm, about 510 nm, about 520 nm, about 530 nm, about 540 nm. about 550 nm, about 560 nm, about 570 nm, about 580 nm, about 590 nm, about 600 nm, about 610 nm, about 620 nm, about 630 nm, about 640 nm, about 650 nm, about 660 nm, about 670 nm, about 680 nm, about 690 nm, about 700 nm, about 710 nm, about 720 nm, about 730 nm, about 740 nm, about 750 nm, about 760 nm, about 770 nm, about 780 nm. about 790 nm, about 800 nm, about 810 nm, about 820 nm. about 830 nm, about 840 nm, about 850 nm, about 860 nm, about 870 nm, about 880 nm, about 890 nm, about 900 nm, or a range between any two of these values. With respect to the above peak absorbance, dyes that have an absorbance wavelength, could be + / - 10 nm, + / - 3 nm, + / - 2 nm, or + / - 1 nm, or + / - 10%, + / - 5%, + / - 1% in the context of the compounds and methods herein.

[0329] In some embodiments, the dyes disclosed herein have a peak emission of about 300 nm, about 310 nm, about 320 nm, about 330 nm, about 340 nm, about 350 nm, about 360 nm, about 370 nm, about 380 nm, about 390 nm, about 400 nm, about 410 nm, about 420 nm, about 430 nm, about 440 nm, about 450 nm, about 460 nm, about 470 nm, about 480 nm, about 490 nm, about 500 nm, about 510 nm, about 520 nm, about 530 nm. about 540 nm, about 550 nm, about 560 nm, about 570 nm, about 580 nm, about 590 nm, about 600 nm, about 610 nm, about 620 nm, about 630 nm, about 640 nm, about 650 nm, about 660 nm, about 670 nm, about 680 nm, about 690 nm, about 700 nm, about 710 nm, about 720 nm, about 730 nm, about 740 nm, about 750 nm, about 760 nm, about 770 nm. about 780 nm, about 790 nm, about 800 nm, about 810 nm, about 820 nm, about 830 nm, about 840 nm, about 850 nm, about 860 nm, about 870 nm, about 880 nm, about 890 nm, about 900 nm, about 910 nm, about 920 nm, about 930 nm, about 940 nm, about 950 nm, about 960 nm, about 970 nm, about 980 nm, about 990 nm, about 1000 nm, or a range between any two of these values. With respect to the above peak emission, dyes that have an emission wavelength, could be + / - 10 nm, + / - 3 nm, + / - 2 nm, or + / - 1 nm, or + / -10%, + / - 5%, + / - 1% in the context of the compounds and methods herein.

[0330] It is understood that the absorption spectra of fluorophores and fluorescent dyes may vary when conjugated to a peptide, or in a composition or milieu for delivery, or in the body of a subject, and one of skill in the art would appreciate that any of the foregoing absorption values of the fluorophore or dye could be + / - 10 nm, + / - 3 nm, + / - 2 nm, or + / - 1 nm, or + / - 10%, + / -5%, + / - 1% in the context of the compounds and methods herein. In some embodiments, depending on the environment that the fluorophore molecule is in, the fluorophore molecule hasan optimal excitation spectrum from 600 nm to 900 nm. Some other exemplary dyes used in the present disclosure can include near-infrared dyes, such as, but not limited to, Dy Light-680, DyLight-750, VivoTag-750, DyLight-800, IRDye-800, VivoTag-680, Cy5, Cy5.5, Cy7, Cy7.5, or an indocyanine green (ICG). In some aspects, near infrared dyes often include cyanine dyes. Additional non-limiting examples of fluorescent dyes for use as a conjugating molecule in the present disclosure can include acradine orange or yellow, Alexa Fluors and any derivative thereof, 7-actinomycin D, 8-anilinonaphthalene-l -sulfonic acid, ATTO dye and any derivative thereof, auramine-rhodamine stain and any derivative thereof, benzanthrone, bimane, 9-10-bis(phenylethynyl)anthracene. 5,12 - bis(phenylethynyl)naththacene. bisbenzimide, brainbow. calcein, carboxyfluorescein and any derivative thereof, 1 -chi oro-9, 10-bis(phenylethynyl)anthracene and any derivative thereof, DAPI, DiOC6, DyLight Fluors and any derivative thereof, epicocconone, ethidium bromide, FlAsH-EDT2, Fluo dye and any derivative thereof, FluoProbe and any derivative thereof, Fluorescein and any derivative thereof. Fura and any derivative thereof, GelGreen and any derivative thereof, GelRed and any derivative thereof, fluorescent proteins and any derivative thereof, m isoform proteins and any derivative thereof such as for example mCherry. heptamethine dye and any derivative thereof, hoeschst stain, iminocoumarin, indian yellow, indo-1 and any derivative thereof, laurdan, lucifer yellow and any derivative thereof, luciferin and any derivative thereof, luciferase and any derivative thereof, mercocyanine and any derivative thereof, nile dyes and any derivative thereof, perylene, phloxine, phyco dye and any derivative thereof, propidium iodide, pyranine, rhodamine and any derivative thereof, ribogreen, RoGFP, rubrene, stilbene and any derivative thereof, sulforhodamine and any derivative thereof, SYBR and any derivative thereof, synapto-pHluorin, tetraphenyl butadiene, tetrasodium tris, Texas Red, Titan Yellow, TSQ, umbelliferone, violanthrone, yellow fluroescent protein, YOYO-1 and ZW800. Other suitable fluorescent dyes include, but are not limited to, fluorescein and fluorescein dyes (e.g., fluorescein isothiocyanate or FITC, naphthofluorescein, 4', 5' -dichloro-2',7' -dimethoxyfluorescein, 6-carboxyfluorescein or FAM, etc.), carbocyanine, merocyanine, styryl dyes, oxonol dyes, phycoerythrin, erythrosin, eosin, rhodamine dyes (e.g., carboxytetramethyl-rhodamine or TAMRA, carboxyrhodamine 6G, carboxy-X-rhodamine (ROX), lissamine rhodamine B, rhodamine 6G, rhodamine Green, rhodamine Red, tetramethylrhodamine (TMR), etc.), coumarin and coumarin dyes (e.g., methoxycoumarin, dialkylaminocoumarin, hydroxy coumarin, aminomethylcoumarin (AMCA), etc.), Oregon Green Dyes (e.g., Oregon Green 488, Oregon Green 500, Oregon Green 514., etc ), Texas Red, Texas Red-X, SPECTRUM RED, SPECTRUM GREEN, cyanine dyes (e.g., CY-3, Cy-5, CY-3.5, CY-5.5, etc ), ALEXA FLUOR dyes (e g., ALEXA FLUOR 350, ALEXAFLUOR 488, ALEXA FLUOR 532, ALEXA FLUOR 546, ALEXA FLUOR 568, ALEXA FLUOR 594, ALEXA FLUOR 633, ALEXA FLUOR 660, ALEXA FLUOR 680, etc ), BODIPY dyes (e g., BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650. BODIPY 650 / 665. etc.), IRDyes (e.g.. IRD40. IRD 700, IRD 800, etc.), and the like.

[0331] In some aspects, the indocyanine green (ICG) has a peak absorbance of about 600 nm to about 900 nm.

[0332] In some aspects, the indocyanine green (ICG) has a peak absorbance of 750 nm-770 nm, 755 nm-775 nm, or 768nm.

[0333] In some aspects, the indocyanine green (ICG) has a peak emission of about 750 nm to about 950 nm.

[0334] In some aspects, the indocyanine green (ICG) has a peak emission of about 790 nm-820 nm, 805 nm-815 nm. or 807nm.

[0335] In some aspects, the techniques described herein relate to a method, wherein the fluorescent dye on the polypeptide includes DyLight-680, DyLight-750, VivoTag-750, DyLight-800, IRDye-800, VivoTag-680, Cy5, Cy5.5, Cy7, Cy7.5, an indocyanine green (ICG), near infrared dyes, acradine orange or yellow, 7-actinomycin D, 8-anilinonaphthalene-l-sulfonic acid, ATTO dye and any derivative thereof, auramine-rhodamine stain and any derivative thereof, benzanthrone, bimane, 9-10-bis(phenylethynyl)anthracene, 5,12 -bis(phenylethynyl)naththacene, bisbenzimide, brainbow, calcein, carboxyfluorescein and any derivative thereof, 1 -chi oro-9, 10-bis(phenyIethynyI)anthracene and any derivative thereof, DAPI, DiOC6. Dy Light Fluors and any derivative thereof, epicocconone, ethidium bromide. FlAsH-EDT2, Fluo dye and any derivative thereof, FluoProbe and any derivative thereof, Fluorescein and any derivative thereof, Fura and any derivative thereof, GelGreen and any derivative thereof, GelRed and any derivative thereof, fluorescent proteins and any derivative thereof, m isoform proteins and any derivative thereof, heptamethine dye and any derivative thereof, hoeschst stain, iminocoumarin, indian yellow, indo-1 and any derivative thereof, laurdan, lucifer yellow and any derivative thereof, luciferin and any derivative thereof, luciferase and any derivative thereof, mercocyanine and any derivative thereof, methylene blue and any derivative thereof, nile dyes and any derivative thereof, OS680, OS750, perylene, phloxine, phyco dye and any derivative thereof, propidium iodide, pyranine, rhodamine and any derivative thereof, ribogreen, RoGFP, rubrene, stilbene and any derivative thereof, sulforhodamine and any derivative thereof, SYBR and any derivative thereof, synapto-pHluorin, tetraphenyl butadiene, tetrasodium tris, Titan Yellow, topotecan, TSQ, umbelliferone, violanthrone, yellow fluroescentprotein, YOYO-1 and ZW800, fluorescein and fluorescein dyes, carbocyanine, merocyanine, styryl dyes, oxonol dyes, phycoerythrin, erythrosin, eosin, rhodamine dyes, coumarin and coumarin dyes, Oregon Green Dyes, Texas Red, Texas Red-X, SPECTRUM RED, SPECTRUM GREEN, cyanine dyes, ALEXA FLUOR dyes and any derivative thereof, BODIPY dyes, IRDyes, or any combination thereof.

[0336] The present disclosure also relates to a method of imaging an abnormal tissue, cancer, tumor, vasculature, nerves, or structure from a subject using a detectable agent (e.g., a fluorophore) in accordance with any of the disclosed methods. The disclosed methods further include administering a contrast or imaging agent to the subject, and producing an image of the abnormal tissue, cancer, tumor, vasculature, nerve, or structure by imaging the contrast or imaging agent using an imaging system. In an aspect, the contrast or imaging agent includes a dye, a fluorophore, a fluorescent biotin compound, a luminescent compound, a chemiluminescent compound, nerve imaging compound, or any combination thereof.

[0337] In various aspects of the disclosed methods, the contrast or imaging agent further includes a protein, peptide, amino acid, nucleotide, polynucleotide, or any combination thereof. In an aspect, the contrast or imaging agent further includes tozuleristide (also referred to herein as “Compound 76"’). In another aspect, the contrast or imaging agent absorbs a wavelength between from about 200 nm to about 900 nm. In other aspects, the contrast or imaging agent includes DyLight-680, DyLight-750, VivoTag-750, DyLight-800, IRDye-800, VivoTag-680, Cy5, Cy5.5, Cy7, Cy7.5, or an indocyanine green (ICG) and any derivative of the foregoing; fluorescein and fluorescein dyes (e.g., fluorescein isothiocyanate or FITC, naphthofluorescein, 4', 5'-dichloro-2',7' -dimethoxyfluorescein, 6-carboxyfluorescein or FAM, etc.), carbocyanine, merocyanine, styryl dyes, oxonol dyes, phycoerythrin, rythrosine, eosin, rhodamine dyes (e.g., carboxytetramethyl-rhodamine or TAMRA, carboxyrhodamine 6G, carboxy -X-rhodamine (ROX), lissamine rhodamine B, rhodamine 6G, rhodamine Green, rhodamine Red, tetramethylrhodamine (TMR), etc. ), coumarin, coumarin dyes (e.g., methoxy coumarin, dialkylaminocoumarin, hydroxy coumarin, aminomethylcoumarin (AMCA), etc.), Oregon Green Dyes (e.g., Oregon Green 488, Oregon Green 500, Oregon Green 514., etc.), Texas Red, Texas Red-X, SPECTRUM RED, SPECTRUM GREEN, cyanine dyes (e.g., CY-3, Cy-5, CY-3.5, CY-5.5, etc. ), ALEXA FLUOR dyes (e g., ALEXA FLUOR 350. ALEXA FLUOR 488, ALEXA FLUOR 532. ALEXA FLUOR 546, ALEXA FLUOR 568, ALEXA FLUOR 594, ALEXA FLUOR 633, ALEXA FLUOR 660, ALEXA FLUOR 680, etc ), BODIPY dyes (e.g., BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650, BODIPY 650 / 665,etc.), IRDyes (e.g, IRD40, IRD 700, IRD 800, etc.), 7-aminocoumarin, a dialkylaminocoumarin reactive dye, 6,8-difluoro-7-hydroxycoumarin fluorophore, a hydroxy coumarin derivative, an alkoxy coumarin derivatives, a succinimidyl ester, a pyrene succinimidyl ester, a pyridyloxazole derivative, an aminonaphthalene-based dyes, dansyl chlorides, a dapoxyl dye, Dapoxyl sulfonyl chloride, amine-reactive Dapoxyl succinimidyl ester, carboxylic acid-reactive Dapoxyl (2-aminoethyl)sulfonamide), a bimane dye, bimane mercaptoacetic acid, an NBD dye, a QsY 35, or any combination thereof.

[0338] The present disclosure also relates to a method of imaging an abnormal tissue, cancer, tumor, vasculature or structure in a fluorophore from a subject in accordance with any of the disclosed methods. The disclosed methods further include administering a contrast or imaging agent to the subject, and producing an image of the abnormal tissue, cancer, tumor, vasculature or structure by imaging the contrast or imaging agent using an imaging system. In an aspect, the contrast or imaging agent includes a dye, a fluorophore. a fluorescent biotin compound, a luminescent compound, a chemiluminescent compound, or any combination thereof.

[0339] In various aspects of the disclosed methods, the contrast or imaging agent further includes a protein, peptide, amino acid, nucleotide, polynucleotide, or any combination thereof. In an aspect, the contrast or imaging agent further includes tozuleristide (Compound 76). In another aspect, the contrast or imaging agent absorbs a wavelength between from about 200 nm to about 900 nm. In other aspects, the contrast or imaging agent includes DyLight-680, DyLight-750, VivoTag-750, DyLight-800, IRDye-800, VivoTag-680, Cy5, Cy5.5, Cy7, Cy7.5, or an indocyanine green (ICG) and any derivative of the foregoing; fluorescein and fluorescein dyes (e.g., fluorescein isothiocyanate or FITC, naphthofluorescein, 4', 5'-dichloro-2',7' -dimethoxyfluorescein, 6-carboxyfluorescein or FAM, etc.), carbocyanine, merocyanine, styryl dyes, oxonol dyes, phycoerythrin, rythrosine, eosin, rhodamine dyes (e.g., carboxy tetramethy 1-rhodamine or TAMRA, carboxyrhodamine 6G, carboxy-X-rhodamine (ROX), lissamine rhodamine B, rhodamine 6G, rhodamine Green, rhodamine Red, tetramethylrhodamine (TMR), etc. ), coumarin, coumarin dyes (e.g., methoxy coumarin, dialkylaminocoumarin.hydroxy coumarin, aminomethyl coumarin (AMCA), etc.), Oregon Green Dyes (e.g., Oregon Green 488, Oregon Green 500, Oregon Green 514., etc.), Texas Red, Texas Red-X, SPECTRUM RED, SPECTRUM GREEN, cyanine dyes (e g., CY-3, Cy-5. CY-3.5, CY-5.5, etc ). ALEXA FLUOR dyes (e g., ALEXA FLUOR 350, ALEXA FLUOR 488, ALEXA FLUOR 532, ALEXA FLUOR 546, ALEXA FLUOR 568, ALEXA FLUOR 594, ALEXA FLUOR 633, ALEXA FLUOR 660, ALEXA FLUOR 680, etc ), BODIPY dyes (e g, BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570,BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650, BODIPY 650 / 665, etc ), IRDyes (e g., IRD40, IRD 700, IRD 800, etc.), 7-aminocoumarin, a dialkylaminocoumarin reactive dye, 6,8-difluoro-7-hydroxy coumarin fluorophore, a hydroxy coumarin derivative, an alkoxy coumarin derivatives, a succinimidyl ester, a pyrene succinimidyl ester, a pyridyloxazole derivative, an aminonaphthalene-based dyes, dansyl chlorides, a dapoxyl dye. Dapoxyl sulfonyl chloride, amine-reactive Dapoxyl succinimidyl ester, carboxylic acid-reactive Dapoxyl (2-aminoethyl)sulfonamide), a bimane dye, bimane mercaptoacetic acid, an NBD dye, a QsY 35, or any combination thereof.

[0340] In some aspects, complexes of the present disclosure comprise other dyes, including but not limited to those provided below in TABLE 1. Regarding TABLE 1, the peak absorption and emission values for a given fluorophore can vary depending on the environment (e.g. solution, tissue, etc.) that the fluorophore is present in as well as the concentration of fluorophore or fluorophore conjugate utilized.TABLE 1 - Exemplary Fluorescent Reporter Molecules with Peak Absorbance (Abs.) and Emission (Em.) Wavelengths Specified (nm)

[0341] In some other aspects, the conjugate compounds used include a chemiluminescent compound, colloidal metal, luminescent compound, phosphorescent compound, enzyme, radioisotope, nanoparticle, or paramagnetic labels.

[0342] In certain aspects, complexes used in the present disclosure can be conjugated, complexed, or associated with a microscopic particle or nanoparticle (also referred to as nanopowder or nanocluster or nanocrystal) as a detectable agent or therapeutic agent. Such complexes employing nanoparticles can be used to deliver drugs, heat, light or other substances to specific types of tissues and cells (such as cancer cells or vascular lesions) and reduces damage to healthy cells in the body and allows for earlier detection of disease. Exemplary nanoparticle compositions have been used in preclinical nuclear imaging of cardiac and vascular structures, include but are not limited to micelles, liposomes, polymeric particles, dendrimers, lipoprotein particles, gold particles, iron oxide particles, perfluorocarbon emulsions, carbon nanotubes, and up conversion nanophosphors.

[0343] In certain aspects, the complexes used in the present disclosure can be conjugated to radioactive isotopes instead of or in addition to other ty pes of detectable agents. Certain isotopes suitable for use in the present compounds can include, but are not limited to, iodine- 131, iodine-125, bismuth-212, bismuth-213. lutetium-177, rhenium-186, rhenium-188, yttrium-90. astatine-211, phosphorus-32 and / or samarium-153. In some aspects, the complexes of the present disclosure contain one or more atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature, including but not limited to hydrogen, carbon, fluorine, phosphorous, copper, gallium, yttrium, technetium, indium, iodine, rhenium, thallium, bismuth, astatine, samarium, and lutetium (for example,3H,2H,13C,14C,18F,32P,35S,64Cu,67Ga,90Y,99MTC,mIn,125I,123I,131I,135I,186Re,187Re,2O1T1,212BI,211At,153Sm and / or177LU). In other aspects, the complexes of the present disclosure are labeled with a paramagnetic metal ion that is a good contrast enhancer in Magnetic Resonance Imaging (MRI). Examples of such paramagnetic metal ions include, but are not limited to, gadolinium III (Gd3+), chromium 111 (Cr3+), dysprosium III (Dy3+), iron 111 (Fe3+), manganese II (Mn2+), and ytterbium III (Yb3+). In certain embodiments, the labeling moiety' comprises gadolinium III (Gd3+).

[0344] In some aspects, the complexes used in the present disclosure can be conjugated to biotin. In addition of extension of half-life, biotin can also act as an affinity handle for retrieval of the peptides from tissues or other locations. In one aspect, the complexes are conjugated, e.g., to a biotinidase resistant biotin with a PEG linker (e.g., NHS-dPEG4-Biotinidase resistant biotin). In some aspects, fluorescent biotin complexes that can act both as a detectable label andan affinity handle are used. Non-limiting examples of commercially available fluorescent biotin complexes can include Atto 425-Biotin, Atto 488-Biotin, Atto 520-Biotin, Atto-550 Biotin, Atto 565-Biotin, Atto 590-Biotin, Atto 610-Biotin, Atto 620-Biotin, Atto 655-Biotin, Atto 680-Biotin, Atto 700-Biotin, Atto 725-Biotin, Atto 740-Biotin, fluorescein biotin, biotin-4-fluorescein, biotin-(5-fluorescein) conjugate, and biotin-B-phycoerythrin, alexa fluor 488 biocytin, alexa fluor 546, alexa fluor 549, lucifer yellow cadaverine biotin-X, Lucifer yellow biocytin, Oregon green 488 biocytin, biotin-rhodamine, and tetramethylrhodamine biocytin.

[0345] For example, human serum albumin (HSA) can be conjugated to the tumor homing agent or the nerve homing agent the present invention and thereby increase retention within the cancer cell or vasculature and its half-life. Peptides, antibodies, or antibody fragments can be engineered to target specific tissues of interest, for example cancer cell, vascular endothelium, or nerves, so that these structures are stably labeled for the duration of a surgical or diagnostic procedure. Complexes can be created that are non-fluorescent until they are activated in the presence of the diseased tissue or other condition to be detected. Examples include peptide moieties that are cleaved by cathepsins or matrix metalloproteinases that can be used to detect vascular lesions including areas of abnormal tissue or inflammation.

[0346] In certain embodiments, the peptide and peptide variants can be conjugated to moieties, such as detectable labels (e.g., dyes) that can be detected (e.g., visualized) in a subject. In some embodiments, the peptide and / or peptide variants can be conjugated to detectable labels to enable tracking of the bio-distribution of a conjugated peptide. The detectable labels can include fluorescent dyes. Non-limiting examples of fluorescent dyes that can be used as a conjugating molecule in the present disclosure include rhodamine, rhodol. fluorescein, thiofluorescein, aminofluorescein, carboxyfluorescein, chlorofluorescein, methylfluorescein, sulfofluorescein, aminorhodol, carboxyrhodol, chlororhodol, methylrhodol, sulforhodol; aminorhodamine, carboxyrhodamine, chlororhodamine, methylrhodamine, sulforhodamine, and thiorhodamine, cyanine, indocarbocyanine, oxacarbocyanine, thiacarbocyanine, merocyanine, a cyanine dye (e.g.. cyanine 2, cyanine 3, cyanine 3.5, cyanine 5, cyanine 5.5. cyanine 7). oxadiazole derivatives, pyridyloxazole, nitrobenzoxadi azole, benzoxadiazole, pyrene derivatives, cascade blue, oxazine derivatives, Nile red, Nile blue, cresyl violet, oxazine 170, acridine derivatives, proflavin, acridine orange, acridine yellow, arylmethine derivatives, auramine, xanthene dyes, sulfonated xanthenes dyes. Alexa Fluors (e.g.. Alexa Fluor 594, Alexa Fluor 633, Alexa Fluor 647, Alexa Fluor 700), crystal violet, malachite green, tetrapyrrole derivatives, porphyrin, phtalocyanine, and bilirubin. Some other example dyes include near-infrared dyes, such as, butnot limited to, Cy5, Cy5.5, Cy7, Cy7.5, an indocyanine green (ICG), DyLight 750 or IRdye 800. In some embodiments, near infrared dyes can include cyanine dyes.

[0347] In other embodiments, therapeutic agents, anti-cancer drugs, and anti-vascular lesion drugs, and agents, include, but are not limited to: radioisotopes, nanoparticle, toxins, enzymes, sensitizing drugs, radiosensitizers, photosensitizers, nucleic acids, including interfering RNAs. antibodies, antibody fragments, anti-angiogenic agents, aptamer, anti-angiogenic agent, antimetabolite, mitotic inhibitor, growth factor inhibitor, anti-metabolites, mitotic inhibitors, growth factor inhibitors, and their equivalents, as well as photo-ablation.

[0348] As used herein, the terms "about” and “approximately,” in reference to a number, is used herein to include numbers that fall within a range of 10%, 5%, or 1% in either direction (greater than or less than) the number unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0349] Suitable diagnostic agents can include agents that provide for the detection by fluorescence methods as well as methods other than fluorescence imaging. Other suitable diagnostic agents can include radiolabels (e.g., radio isotopically labeled compounds) such as125I,14C, and31P, among others; and magnetic resonance imaging agents.

[0350] Suitable targeting agents can include antibodies, polypeptides, polysaccharides, nucleic acids, fatty acids, lipids, glycolipids, sterols, vitamins, cofactors, hormones, neurotransmitters, and metabolites.

[0351] In another aspect of the invention, compositions used can include the peptide complexes as provided. In yet another aspect of the invention, compositions used can include peptide complexes as discussed herein. The composition used can include a pharmaceutically acceptable carrier or diluent for deliver}' of the peptide conjugate. Suitable pharmaceutically acceptable carriers or diluents can include saline or dextrose for injection.

[0352] In various aspects, the presently described compounds used can further comprise a detectable label, which can be used for the detection of the peptide-label conjugate and the tissue or cells or vascular lesion tissues or cells to which they are bound or accumulated.

[0353] In various aspects, compounds used in the present disclosure can have the structure of Formula (I), or a pharmaceutically acceptable salt thereof:wherein:R1, R2, R3, R4, R5, R6, R7, R8, R13, and R16are each independently selected from hydrogen, Ci-Ce alkyl. Ci-Cs alkylene-COOH, sulfonate. -COOH, -SO2-NH2. Ci-Cs alkoxy, C1-C10 alkylene-(C (= O))x- C1-C10 alkylene-(C (= O))x-O- or C1-C10 alkylene-(C (= O))x-NR10-;R9is hydrogen, sulfonate, -COOH, C1-C10 alkylene-(C (= O))x-, C1-C10 alkylene-(C (= O))x-O-. or C1-C10 alkylene-(C (= O))X-NR10-;L1is C3-C6 alkylene;L2is C1-C10 alkylene;L3is a bond, -O-, -NR10-, -NR10-CI-C6 alkylene-, -O-NR10-, -NR10-Ci-Cs alkylene-(O-C1-C6 alkylene)n-, -NR10-L4-. -NR10-CI-C6 alkylene-NR11- (C ( = O) -Ci-Ce alkylene-O- )m-, or-NR10-Ci-Ce alkylene-NR10-Ci-C6 alkylene-NR10-Ci-C6 alkylene-;L4is a bond, -heterocyclyl-, or -heterocyclyl-Ci-Cs alkylene-;R10is hydrogen or Ci-Ce alkyl;R11is hydrogen or Ci-Ce alkyl;R12and R13are each independently selected from hydrogen. Ci-Ce alkyl, or R12and R13are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R14is hydrogen or Ci-Ce alkylene, -(L5)-aryl, -(L5)-aryl-A5, -(L5)-heteroaryl, — (L5)— heteroaryl-A5, -NR17R18, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;L5is a bond, C1-C10 alkylene, -O-, or -NR10-;R17and R18are each independently hydrogen or aryl;R19and R20are each independently selected from hydrogen, Ci-Ce alkyl, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocy clic or heterocy clic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;n is 0, 1, 2, or 3;m is 0, 1, 2, or 3;p is 0, 1, 2, or 3;q is 0, 1, 2, or 3;x is 0 or 1; andone of A1, A2, A3, A4, or A5is a polypeptide having at least 85% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof and the others of A1, A2, A3, A4, or A5are each independently absent, hydrogen, -COOH, or sulfonate.

[0354] In various aspects, the presently described compounds used can further comprise a detectable label, which can be used for the detection of the peptide-label conjugate and the vascular lesion tissues or cells to which they are bound or accumulated.

[0355] In various aspects, compounds used in the present disclosure have the structure of Formula (II), or a pharmaceutically acceptable salt thereof:wherein:R3, R4, R5, R6, R15, and R16are each independently selected from hy drogen, Ci-Ce alkyl, Ci-Ce alkylene-COOH, sulfonate, -COOH, -SO2-NH2, Ci-Ce alkoxy, C1-C10 alkylene-(C (= O))x-, C1-C10 alkylene-(C (= O))x-O-, or C1-C10 alkylene-(C (= O))x-NR10-;R9is hydrogen, sulfonate, -COOH, C1-C10 alkylene-(C (= O))x—, C1-C10 alkylene-(C (= O))x-O-, or C1-C10 alkylene-(C (= O))X-NR10-;L1is C3-C6 alkylene;L2is C1-C10 alkylene;L3is a bond, -O-, -NR10-, -NR10-Ci-Ce alkylene-, -O-NR10-, -NR10-Ci-Ce alkylene-(O-Ci-Ce alkylene)n-, -NR10-L4-, -NR10-CI-C6alkylene-NR11- (C ( = O) -Ci-C6alkylene-O-)m-, or-NR10-Ci-Ce alkylene-NR10-Ci-C6 alkylene-NR10-Ci-C6 alkylene-;L4is a bond, -heterocyclyl-, or -heterocyclyl-Ci-Cs alkylene-;R10is hydrogen or Ci-Ce alkyl;R11is hydrogen or Ci-Ce alkyl;R12and R13are each independently selected from hydrogen. Ci-Ce alkyl, or R12and R13are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R14is hydrogen or Ci-Ce alkylene, -(L5)-aryl, -(L5)-aryl-A5, -(L5)-heteroaryl, — (L5)— heteroaryl-A5, -NR17R18, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;L5is a bond, C1-C10 alkylene, -O-, or -NR10-;R17and R18are each independently hydrogen or aryl;R19and R20are each independently selected from hydrogen. Ci-Ce alkyl, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R21and R22are each independently selected from hydrogen, Ci-Ce alkyl, sulfonate, or R21and R22are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered aryl;R23and R24are each independently selected from hydrogen. Ci-Ce alkyl, sulfonate, or R23and R24are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered aryl;n is 0, 1, 2, or 3;m is 0, 1, 2, or 3;p is 0, 1, 2, or 3;q is 0, 1, 2, or 3;x is 0 or 1; andone of A1, A2, A3, A4, or A5is a polypeptide having at least 85% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof and the others of A1, A2, A3, A4, or A5are each independently absent, hydrogen, -COOH, or sulfonate.

[0356] In some aspects, the compounds used in the present disclosure have a structure of Formula (III), or a pharmaceutically acceptable salt thereof:(III).

[0357] In certain aspects, the present compounds have a structure of Formula (IV), or a pharmaceutically acceptable salt thereof:wherein:R1, R2, R3, R4, R5, R6, R7, R8, R15, and R16are each independently selected from hydrogen, Ci-Ce alky l. Ci-Ce alkylene-COOH, sulfonate, Ci-Ce alkylene-sulfonate, -COOH. -SO2-NH2, or Ci-Ce alkoxy;R9is hydrogen, sulfonate, amine or -COOH;L1is C3-C6 alky lene;L2is C1-C10 alkylene;L3is a bond, -O-, -NR10-, -NR10-CI-C6 alkylene-, -O-NR10-, -NR10-CI-C6 alkylene-(O-C1-C6 alkylene)n-, -NR10-L4-, -NR10-CI-C6alkylene-NR11- (C (= O) -Ci-C6alkylene-O-)m-, or-NR10-Ci-C6 alkylene — NR10-Ci-Ce alkylene — NR10-Ci-Ce alkylene-;L4is a bond, -heterocyclyl-, or -heterocyclyl-Ci-Cs alkylene-;R10is hydrogen or Ci-Ce alkyl;R11is hydrogen or Ci-Ce alkyl;R12and R13are independently selected from hydrogen, Ci-Ce alkyl, or R12and R13are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R14is hydrogen or Ci-Ce alkylene, -(L5)-aryl, -(L5)-aryl-R21, -(L5)-heteroaryl, — (L5)— heteroaryl-R21, -NR17R18, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;L? is a bond, C1-C10 alkylene, -0- -NR10-;R17and R18are each independently hydrogen or aryl;R19and R20are independently selected from hydrogen, Ci-Ce alkyl, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R21is hydrogen, sulfonate, or -C00H;n is 0, 1, 2, or 3;m is 0, 1, 2, or 3;p is 0, 1, 2, or 3;q is 0, 1, 2, or 3; andA4is a polypeptide having at least 80% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0358] In other aspects, compounds used in the present disclosure have a structure of Formula (V), or a pharmaceutically acceptable salt thereof:wherein:R1, R2, R4, R3, R6, R7, R8, R15, and R16are each independently selected from hydrogen, Ci-Ce alkyl, Ci-Ce alkylene-COOH, sulfonate, -COOH, -SO2-NH2, or Ci-Ce alkoxy;R3is selected from C1-C10 alkylene-(C (= O))x-, C1-C10 alkylene-(C (= O))x-O-, or Ci-C10 alkylene-(C (= O))X-NR10-;R9is hydrogen, sulfonate, or -COOH, or C1-C10 alkyl;L1is Cs-Ce alkylene;L2is C1-C10 alkylene;L3is hydrogen, sulfonate, -COOH, C1-C10 alkyl;L4is a bond, -heterocyclyl-. or -heterocyclyl-Ci-Cs alkylene-;R10is hydrogen or Ci-Ce alkyl;R11is hydrogen or Ci-Ce alkyl;R12and R13are independently selected from hydrogen, Ci-Ce alkyl, or R12and R13are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R14is hydrogen or Ci-Ce alkylene, -(L’)-aryl, -(I?)-heteroaryl, -NR17R18, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;I is a bond, C1-C10 alkylene, -O-, -NR10-;R17and R18are each independently hydrogen or aryl;R19and R20are independently selected from hydrogen, Ci-Ce alkyd. R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with theother atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;n is 0, 1, 2, or 3;m is 0, 1, 2, or 3;p is 0, 1, 2. or 3;q is 0, 1, 2, or 3;x is 0 or 1; andA1is a polypeptide having at least 85% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0359] In other aspects, compounds used in the present disclosure have a structure of Formula (VI), or a pharmaceutically acceptable salt thereof:wherein:R1, R2, R3, R4, R6, R7, R8, R15, and R16are each independently selected from hydrogen, Ci-Ce alkyl, Ci-Ce alkylene-COOH, sulfonate, -COOH, -SO2-NH2, or Ci-Ce alkoxy;R5is selected from C1-C10 alkylene-(C (= O))x-, C1-C10 alkylene-(C (= O))x-O-, or Ci-C10 alkylene-(C (= O))X-NR10-;R9is hydrogen, sulfonate, or -COOH, or C1-C10 alkyl;L1is Cs-Ce alkylene;L2is C1-C10 alkylene;L3is hydrogen, sulfonate, -COOH, or C1-C10 alkyl;L4is a bond, -heterocyclyl-. or -heterocyclyl-Ci-Cs alkylene-;R10is hydrogen or Ci-Ce alkyl;R11is hydrogen or Ci-Ce alkyl;R12and R13are independently selected from hydrogen, Ci-Ce alkyd, or R12and R13are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocy clic or heterocy clic ring;R14is hydrogen or Ci-Ce alkylene, -(L5)-aiy 1, -(L5)-heteroaryl, -NR17R18, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;I? is a bond, C1-C10 alkylene, -O-, -NR10-;R17and R18are each independently hydrogen or aryl;R19and R20are independently selected from hydrogen, Ci-Ce alkyd, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;n is 0, 1, 2, or 3;m is 0, 1, 2, or 3;p is 0, 1, 2, or 3;q is 0, 1, 2, or 3;x is 0 or 1; andA2is a polypeptide having at least 85% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0360] In some aspects, compounds used in the present disclosure have a structure of Formula (VII), or a pharmaceutically acceptable salt thereof:R1, R2, R3, R4, R5, R6, R7, R8, R13, and R16are each independently selected from hydrogen, Ci-Ce alkyl, Ci-Ce alkylene-COOH, sulfonate, -COOH, -SO2-NH2, or Ci-Ce alkoxy;R9is selected from C1-C10 alkylene-(C (= O))x-, C1-C10 alkylene-(C (= O))x-O-, or Ci-C10 alkylene-(C (= O))X-NR10-;L1is C3-C6 alkylene;L2is C1-C10 alkylene;L3is hydrogen, sulfonate, -COOH, or C1-C10 alky l;L4is a bond, -heterocyclyl-, or -heterocyclyl-Ci-C6 alkylene-;R10is hydrogen or Ci-Ce alkyl;R11is hydrogen or Ci-Ce alkyl;R12and R13are independently selected from hydrogen, Ci-Ce alkyd, or R12and R13are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R14is hydrogen or Ci-Ce alkylene, -(L5)-aryl, -(L5)-heteroaryl, -NR17R18, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R17and R18are each independently hydrogen or aryl;R19and R20are independently selected from hydrogen, Ci-Ce alkyd, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;n is 0, 1, 2, or 3;m is 0, 1, 2, or 3;p is 0, 1, 2. or 3;q is 0, 1, 2, or 3;x is 0 or 1;I is a bond, C1-C10 alkylene, -O-, -NR10-;A3is a polypeptide having at least 85% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0361] In additional aspects, compounds used in the present disclosure have a structure Formula (VIII), or a pharmaceutically acceptable salt thereof:wherein:R1, R2, R3, R4, R5, R6, R7, R8, R13, and R16are each independently selected from hydrogen, Ci-Ce alkyl, Ci-Ce alkylene-COOH, sulfonate, -COOH, -SO2-NH2, or Ci-Ce alkoxy;R9is hydrogen, sulfonate, or -COOH;L1is C3-C6 alky lene;L2is C1-C10 alkylene;L3is a bond, -O-, -NR10-. -NR10-Ci-Cs alkylene-, -O-NR10-, -NR10-CI-C6 alkylene-(O-C1-C6 alkylenejn- -NR10-L4-, -NR10-CI-C6alkylene-NR11- (C (= O) -Ci-C6alkylene-O-)m-, or-NR10-Ci-C6 alkylene — NR10-Ci-Ce alkylene — NR10-Ci-Ce alky lene-;L4is a bond, -heterocyclyl-, or -heterocyclyl-Ci-C6 alkylene-;R10is hydrogen or Ci-Ce alkyl;R11is hydrogen or Ci-Ce alkyl;R12and R13are independently selected from hydrogen, Ci-Ce alkyd, or R12and R13are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R14is -(L5)-aryl-A5, or -(L5)-heteroaryl-A5;L3is a bond, C1-C10 alkylene, -O-, -NR10-;R17and R18are each independently hydrogen or ary l;R19and R20are independently selected from hydrogen, Ci-Ce alkyl, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;n is 0, 1, 2, or 3;m is 0, 1, 2, or 3;p is 0, 1, 2, or 3;q is 0, 1, 2, or 3;x is 0 or 1;A4is hydrogen, -COOH. or sulfonate; andA3is a polypeptide having at least 85% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0362] In certain aspects. A1, A2, and A3are absent. In some aspects, A3is hydrogen. In certain aspects, R3, R4, R5, and R6are each independently Ci-Ce alkyl. In some aspects, R3, R4, R5, and R6are each independently methyl. In certain aspects, R1, R2, R7, R8, R15, and R16are each independently selected from hydrogen or sulfonate. In further aspects, R1, R2, R7, R8, R15, and R16are each independently hydrogen. In some aspects, R12, R13, R14, R19, R20are each independently hydrogen.

[0363] In certain aspects, R12and R13join together along with the atoms to which they are attached to form a six-membered carbocyclic ring. In other aspects, R12and R13join together along with the atoms to which they are attached to form a five-membered carbocyclic ring. In certain aspects, R14and R19join together along with the atoms to which they are attached to form a six-membered carbocyclic ring. In some aspects, R14and R20join together along with the atoms to which they are attached to form a six-membered carbocyclic ring. In certain aspects, L1is C3-C6 alkylene. In other aspects, L1is C3-C5 alkylene. In still other aspects, L1is propylene. In still other aspects, L1is buty lene. In other aspects, L1is pentylene. In some aspects, L2is C3-C6 alky lene. In other aspects, L2is propylene. In still other aspects, L2is butylene. In other aspects, L2is pentylene. In some aspects. R9is sulfonate. In other aspects. R9is hydrogen. In some aspects, R14is hydrogen. In other aspects, R14is -(L3)-aryl. In still other aspects, R14is — (L5)— aryl-A5.

[0364] In some aspects, R1is hydrogen. In certain aspects, R2is hydrogen. In some aspects, R3is methyl. In certain aspects, R4is methyl. In some aspects, R3is methyl. In certain aspects R6is methyl. In some aspects, R7is hydrogen. In certain aspects, R8is hydrogen. In some aspects, R12is hydrogen. In certain aspects, R13is hydrogen. In some aspects, R14is hydrogen. In certain aspects, R19is hydrogen. In some aspects, R20is hydrogen. In certain aspects, R10is hydrogen. In some aspects, R11is hydrogen.

[0365] In some aspects, R17and R18are independently phenyl. In some aspects, L1is buytlene. In some aspects, L2is pentylene. In some aspects, L3is selected from a bond, -O-, -NR10-, -NR10-Ci-Ce alkylene-, -O-NR10-, or -NR10-L4-. In further aspects, L3is a bond.

[0366] In some aspects, L4is -heterocyclyl- or-heterocyclyl-Ci-Ce alkylene- In further - N / l— \ aspects, L4is -piperizinyl-(Ci-C6 alky lene)-. In still further aspects, L4is?.

[0367] In some aspects, p is 1. In certain aspects, q is 1.

[0368] In some aspects, the compound used has the structure of any one of Formulas (IX). (X), (XI), (XII), (XIII), (XIV), (XV), or (XVI):

[0369] In some aspects, the compound has the structures of any one of Formulas (IX), (X), (XI). (XII), (XIII). (XIV), (XV). or (XVI), wherein A4is a polypeptide.

[0370] In some aspects, one of A1, A2, A3, A4, or A5is a polypeptide having at least 87% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof. In further aspects, one of A1, A2, A3, A4, or A5is a polypeptide having at least 90% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof. In still further aspects, one of A1, A2, A3, A4, or A5is a polypeptide having at least 92% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof. In still further aspects, one of A1, A2, A3, A4, or A5is a polypeptide having at least 95% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof. In still further aspects, one of A1, A2, A3, A4, or A5is a polypeptide having at least 97% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof. In still further aspects, one of A1. A2. A3. A4, or A5is a polypeptide having 100% sequence identity withMCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof. In still further aspects, one of A1, A2, A3, A4, or A5is a polypeptide having the sequence MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0371] In some aspects, the fragment of A1, A2, A3, A4, or A5has a length of at least 25 ammo acid residues. In further aspects, the fragment of A1, A2, A3, A4, or A5has a length of at least 27 amino acid residues. In still further aspects, the fragment of A1, A2, A3, A4, or A5has a length of at least 29 amino acid residues. In still further aspects, the fragment of A1, A2, A3, A4, or A5has a length of at least 31 amino acid residues. In still further aspects, the fragment of A1, A2, A3, A4, or A5has a length of at least 33 amino acid residues.

[0372] In some aspects, one of A1, A2, A3, A4, or A5is a polypeptide having at least 85% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof. In certain aspects, one of A1, A2, A3, A4, or A5is a polypeptide having at least 85% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof. In some aspects, one of A1, A2, A3, A4, or A5is a polypeptide having at least 85% sequence identity with MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof wherein one of A1, A2, A3, A4, or A5has a sequence selected from SEQ ID NO: 1-SEQ ID NO: 512.

[0373] In some aspect, the polypeptide contains no lysine residues. In some aspects, the polypeptide used comprises at least one lysine amino acid residue. In certain aspects, the polypeptide comprises a single lysine amino acid residue. In some aspects, the polypeptide comprises one, two, or three lysine amino acid residues. In some aspects, the polypeptide comprises a lysine residue at the position corresponding to K-27 of native chlorotoxin. In some aspects, the polypeptide comprises a lysine residue at the position corresponding to K-23 of native chlorotoxin. In some aspects, the polypeptide comprises a lysine residue at the position corresponding to K-15 of native chlorotoxin.

[0374] In some aspects, one or more of the amino acids of the polypeptide used is substituted with a non-naturally occurring amino acid residue. In further aspects the non-naturally occurring amino acid residue is a citrulline amino acid residue. In still further aspects, L3is attached to A4at a citrulline amino acid residue of the polypeptide.

[0375] In some aspects, L3is attached to A4at a lysine amino acid residue of the polypeptide. In certain aspects, L3is attached to A4at the N-terminus of the polypeptide. In some aspects, L3is attached to A4at the C-terminus of the polypeptide. In some aspects, the R3is attached to A1at aly sine amino acid residue of the peptide, a citrulline amino acid residue of the polypeptide, the N-terminus of the polypeptide, or the C-terminus of the polypeptide. In some aspects, the R5is attached to A2at a lysine amino acid residue of the polypeptide, a citrulline amino acid residue of the polypeptide, the N-terminus of the polypeptide, or the C-terminus of the polypeptide. In some aspects, the R9is attached to A3at a lysine amino acid residue of the polypeptide, a citrulline amino acid residue of the polypeptide, the N-terminus of the polypeptide, or the C-terminus of the polypeptide. In some aspects, the aryl is attached to A5at a lysine amino acid residue of the polypeptide, a citrulline amino acid residue of the polypeptide, the N-terminus of the polypeptide, or the C-terminus of the polypeptide.

[0376] In some aspects, the compound used has the structure of any one of compounds 1 to 60 as found in TABLE 2, in which A is a peptide portion and can comprise any of the peptides described herein, such as any one of SEQ ID NO: 1-SEQ ID NO: 512. In other aspects, the compound used has the structure of any one of compounds 1 to 60 as found in TABLE 2, in which A is a peptide fragment and can comprise a fragment of any of the peptides described herein, such as any one of SEQ ID NO: 1-SEQ ID NO: 512. In some embodiments, the fragment of the polypeptide has a length of at least 25 residues.

[0377] In some aspects, the compound used is conjugated to polyethylene glycol (PEG), hydroxy ethyl starch, polyvinyl alcohol, a water soluble polymer, a zwitterionic water soluble polymer, a water soluble poly(amino acid), an albumin derivative, or a fatty acid.

[0378] In some aspects, the polypeptide used has an isoelectric point of from 5.5 to 9.5. In some aspects, the polypeptide has an isoelectric point of from 7.5 to 9.0. In some aspects, the polypeptide has an isoelectric point of from 8.0 to 9.0. In some aspects, the polypeptide has an isoelectric point of from 8.5 to 9.0. In some aspects, the polypeptide is basic and has an isoelectric point of greater than 7.5. In some aspects, the polypeptide has an isoelectric point of about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5. about 7.6, about 7.7, about 7.8, about 7.9. about 8.0, about 8.1, about 8.2, about 8.3, about 8.4. about 8.5, about 8.6. about 8.7, about 8.8, about 8.9, or about 9.0. In other aspects, the polypeptide comprises an isoelectric point of at least 5.5, at least 6.0, at least 6.5, at least 7.0, at least 7.5, at least 8.0, at least 8.5, at least 9.0, or at least 9.5.

[0379] In some aspects, the polypeptide used comprises at least eight cysteine amino acid residues. In some aspects, the polypeptide comprises eight cysteine amino acid residues. In some aspects, the polypeptide comprises four disulfide bonds. In some aspects, the polypeptide comprises from six to seven cysteine amino acid residues. In some aspects, the polypeptidecomprises three disulfide bonds. In some aspects, the polypeptide comprises at least 1 disulfide bond, at least 2 disulfide bonds, at least 3 disulfide bonds, at least 4 disulfide bonds, at least 5 disulfide bonds, or at least 6 disulfide bonds. In some aspects, the spacing between the cysteine amino acid residues in the polypeptide is about the same as in native chlorotoxin. In some aspects, the distribution of charge on the surface of the polypeptide is about the same as in native chlorotoxin.

[0380] In some aspects, the N-terminus of the polypeptide is blocked by acetylation or cyclization.

[0381] In some aspects, one or more of the methionine amino acid residues used is replaced with an amino acid residue selected from isoleucine, threonine, valine, leucine, serine, glycine, alanine, or a combination thereof. In other aspects, one, two, or three methionine residues of the polypeptide are replaced with other amino acids.

[0382] In some aspects, each amino acid of the polypeptide is independently selected as an L- or D-enantiomer.

[0383] In some aspects, an imaging agent is conjugated to a SEQ ID NO: 9 peptide at K27, or any one of SEQ ID NO: 1 - SEQ ID NO: 481 peptide, SEQ ID NO: 482 - SEQ ID NO: 485 peptide, SEQ ID NO: 486 - SEQ ID NO: 512, or a fragment of the foregoing. The compounds, whether peptides or peptide-complexes can act as targeting moieties for one or more of a solid tumor, or other cancer described herein, a cavernoma (a.k.a., cavernous angiomas, cavernous hemangiomas, or cerebral cavernous malformation (CCM)), an arteriovenous malformation (a.k.a., arteriovenous angiomas, arteriovenous hemangiomas, or cerebral arteriovenous malformation (CAM)), an aneurysm (e.g.. including abdominal aortic, thoracic aortic, and cerebral aneurysm), venous malformation, lymphatic malformation, capillary telangiectasia, mixed vascular malformation, or a spinal dural arteriovenous fistula.

[0384] In some aspects, the compound used is capable of passing across the blood brain barrier. In some aspects, the compound used further comprises a therapeutic agent. In some aspects, the polypeptide is conjugated to the therapeutic agent. In some aspects, the compound used further comprises a therapeutic agent attached to A. In further aspects, the therapeutic agent is a cytotoxic agent. In still other aspects, the therapeutic agent comprises a comprises a radioisotope, nanoparticle, toxin, enzyme, sensitizing drug, radiosensitizer, photosensitizer, nucleic acid, interfering RNA. antibody, antibody fragment, aptamer, anti-angiogenic agent, anti-metabolite, mitotic inhibitor, growth factor inhibitor, or a combination thereof.

[0385] In some aspects, the compound of the composition used is any suitable compound described herein. In other aspects, the compound of the composition further comprises an agent.In some aspects, the compound comprises a detectable agent. In one embodiment, the polypeptide is conjugated to an agent. In another embodiment, the polypeptide is conjugated to a detectable agent. In some embodiments, a detectable agent is a detectable label. In some embodiments, a detectable agent comprises a dye. a fluorophore, a fluorescent biotin compound, a luminescent compound, a chemiluminescent compound, a radioisotope, nanoparticle, a paramagnetic metal ion, a tumor homing agent, a nerve homing agent, and a combination thereof. In some embodiments, the polypeptide comprises a single lysine residue and the agent is conjugated to the polypeptide at the single lysine residue. In some embodiments, the polypeptide comprises no lysine residues and the agent is conjugated to the polypeptide at the N-terminus of the polypeptide.

[0386] Certain exemplary compounds falling within the scope of these geneses are provided below in TABLE 2 and further described herein, including both the peptide portion (indicated by A) and the detectable label portion.TABLE 2 - Compounds According to the Present Disclosure

[0387] The peptide portion “A” in compounds 1-60 can comprise any of the peptides described herein, such as any one of SEQ ID NO: 1-SEQ ID NO: 512. In some embodiments, the peptide portion A is SEQ ID NO: 5 attached at K-27 to any one of compounds 1-60. In some embodiments, the peptide portion A is SEQ ID NO: 6 attached at K-27 to any one of compounds 1-60. In some embodiments, the peptide portion A is SEQ ID NO: 8 attached at K-27 to any one of compounds 1-60. In some embodiments, the peptide portion A is SEQ ID NO: 9 attached at K-27 to any one of compounds 1-60. In some embodiments, the peptide portion A is SEQ ID NO: 11 attached at K-23 to any one of compounds 1-60. In some embodiments, the peptide portion A is SEQ ID NO: 12 attached at K-23 to any one of compounds 1-60. In some embodiments, the peptide portion A is SEQ ID NO: 13 attached at K-15 to any one of compounds 1-60. In some embodiments, the peptide portion A is SEQ ID NO: 16 attached at K-15 to any one of compounds 1-60. In some embodiments, the peptide portion A is SEQ ID NO: 20 attached at K-23 to any one of compounds 1-60. In some embodiments, the peptide portion Ais SEQ ID NO: 21 atached at K-23 to any one of compounds 1-60. In some embodiments, the peptide portion A is SEQ ID NO: 22 atached at K-15 to any one of compounds 1-60. In some embodiments, the peptide portion A is SEQ ID NO: 25 atached at K-15 to any one of compounds 1-60.

[0388] TABLE 3 below sets forth certain polypeptide sequences for use with the present disclosure. Citrulline is designated as “Cit” in the sequences.TABLE 3 - Exemplary Peptide Sequence Suitable for Use in the Compounds of the Present Disclosure. Cit = Citrulline.

[0389] Peptide complexes used in this disclosure can comprise a peptide and a labeling agent or detectable label. In an embodiment, peptide is a variant comprising at least 60%, 65%, 70%, 75%, 80%, 83%, 85%, 86%, 89%, 90%, 92%. 93%. 94%. 95%. 96%. 97%. 98%. or 99% identical to the sequence of the native peptide of peptide or a fragment thereof.

[0390] In another embodiment, the compound comprises a polypeptide having at least at least 60%, 65%, 70%, 75%, 80%, 83%, 85%, 86%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512, or any fragment thereof.

[0391] In another embodiment, the present disclosure provides a peptide having the following ammo acid sequence: MCMPCFTTDHQMARKCDDCCGGKGRGKCYGPQCLCR (SEQ ID NO: 1) or a fragment thereof. In a further embodiment, the present disclosure provides peptide variants comprising at least 60%, 65%, 70%, 75%, 80%, 83%, 85%, 86%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the following amino acid sequence: MCMPCFTTDHQMARKCDDCCGGKGRGKCYGPQCLCR (SEQ ID NO: 1) or a fragment thereof.

[0392] In another embodiment, the present disclosure provides a peptide having the following amino acid sequence: MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof. In a further embodiment, the present disclosure provides peptide variants comprising at least 60%, 65%, 70%, 75%, 80%, 83%, 85%, 86%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the following amino acid sequence: MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0393] In a further embodiment, the present disclosure provides peptide variants comprising at least 80%, identical to the following amino acid sequence:MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0394] In a further embodiment, the present disclosure provides peptide variants comprising at least 83% identical to the following amino acid sequence:MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0395] In a still further embodiment, the present disclosure provides peptide variants comprising at least 86% identical to the following amino acid sequence:MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0396] In another embodiment, the present disclosure provides peptide variants comprising at least 88% identical to the following amino acid sequence:MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0397] In a further embodiment, the present disclosure provides peptide variants comprising at least 90% identical to the following amino acid sequence:MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0398] In a still further embodiment, the present disclosure provides peptide variants comprising at least 91% identical to the following amino acid sequence:MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0399] In a still further embodiment, the present disclosure provides peptide variants comprising at least 94% identical to the following amino acid sequence:MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0400] In yet another embodiment, the present disclosure provides peptide variants comprising at least 97% identical to the following amino acid sequence:MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9) or a fragment thereof.

[0401] In another embodiment, the present disclosure provides a peptide having the following amino acid sequence: MCMPCFTTDHQMARXCDDCCGGXGRGXCYGPQCLCR (SEQ ID NO: 482) or a fragment thereof, wherein each X can each independently be any amino acid. In another embodiment, the present disclosure provides a peptide having the following amino acid sequence: MCMPCFTTDHQMARXCDDCCGGXGRGXCYGPQCLCR (SEQ ID NO: 483) or a fragment thereof, wherein X is selected from K, A and R.

[0402] In another embodiment, the chlorotoxin is a peptide or variant thereof having the following ammo acid sequence: MCMPCFTTDHQMARXCDDCCGGXGRGXCYGPQCLCR (SEQ ID NO: 484) or a fragment thereof, wherein each X can independently be R or A.

[0403] In another embodiment, the chlorotoxin is a peptide or variant thereof having the following amino acid sequence: MCMPCFTTDHQMARXCDDCCGGXGRGKCYGPQCLCR (SEQ ID NO: 485) or a fragment thereof, wherein each X can independently be R or A.

[0404] In still other instances, the variant nucleic acid molecules of a peptide of any one of SEQ ID NO: 1 - SEQ ID NO: 512 can be identified by either a determination of the sequence identity of the encoded peptide amino acid sequence with the amino acid sequence of any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512, or by a nucleic acid hybridization assay. Such peptide variants can include nucleic acid molecules (1) that remain hybridized with a nucleic acid molecule having the nucleotide sequence of any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512 (or its complement) under stringent washing conditions, in which the wash stringency is equivalent to 0.5x-2xSSC with 0.1% SDS at 55-65° C, and (2) that encode a peptide having at least 70%, at least 80%, at least 90%, at least 95% or greater than 95% sequence identity to the amino acid sequence of any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512. Alternatively, peptide variants of any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512 can be characterized as nucleic acid molecules (1) that remain hybridized with a nucleic acid molecule having the nucleotide sequence of any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512 (or its complement) under highly stringent washing conditions, in which the wash stringency is equivalent to 0.1x-0.2xSSC with 0.1% SDS at 50-65° C., and (2) that encode a peptide having at least 70%, at least 80%, at least 90%, at least 95% or greater than 95% sequence identity' to the amino acid sequence of any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512.

[0405] The term '‘engineered,” when applied to a polynucleotide, denotes that the polynucleotide has been removed from its natural genetic milieu and is thus free of other extraneous or unwanted coding sequences, and is in a form suitable for use within genetically engineered protein production systems. Such engineered molecules are those that are separated from their natural environment and include cDNA and genomic clones (i.e., a prokaryotic or eukaryotic cell with a vector containing a fragment of DNA from a different organism).Engineered DNA molecules of the present invention are free of other genes with which they are ordinarily associated but may include naturally occurring or non-naturally occurring 5' and 3' untranslated regions such as enhancers, promoters and terminators.

[0406] An ‘‘engineered” polypeptide or protein is a polypeptide or protein that is found in a condition other than its native environment, such as apart from blood and animal tissue. In a preferred form, the engineered polypeptide is substantially free of other polypeptides,particularly other polypeptides of animal origin. It is preferred to provide the polypeptides in a highly purified form, e.g., greater than 95% pure, more preferably greater than 98% pure or greater than 99% pure. When used in this context, the term "engineered" does not exclude the presence of the same polypeptide in alternative physical forms, such as dimers, heterodimers and multimers, or alternatively glycosylated, carboxylated, modified, or derivatized forms.

[0407] An '‘engineered” peptide or protein is a polypeptide that is distinct from a naturally occurring polypeptide structure, sequence, or composition. Engineered peptides include non-naturally occurring, artificial, isolated, synthetic, designed, modified, or recombinantly expressed peptides.

[0408] Polypeptides of the disclosure include polypeptides that have been modified in any way, for example, to: (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) alter binding affinity for forming protein complexes, (4) alter binding affinities, and (5) confer or modify other physicochemical or functional properties. For example, single or multiple amino acid substitutions (e.g., conservative amino acid substitutions) are made in the naturally occurring sequence (e.g., in the portion of the polypeptide outside the domain(s) forming intermolecular contacts). A “conservative amino acid substitution” can refer to the substitution in a polypeptide of an amino acid with a functionally similar amino acid. The following six groups each contain amino acids that can be conservative substitutions for one another: i) Alanine (A), Serine (S), and Threonine (T); ii) Aspartic acid (D) and Glutamic acid (E); iii) Asparagine (N) and Glutamine (Q); iv) Arginine (R) and Lysine (K); v) Isoleucine (I), Leucine (L), Methionine (M). and Valine (V); vi) Phenylalanine (F), Tyrosine (Y), and Tryptophan (W).

[0409] The terms “polypeptide fragment” and “truncated polypeptide” as used herein can refer to a polypeptide that has an amino-terminal and / or carboxy -terminal deletion as compared to a corresponding full-length peptide or protein. In various embodiments, fragments are at least 5, at least 10, at least 25, at least 50, at least 100, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 600, at least 700, at least 800, at least 900 or at least 1000 amino acids in length. In various embodiments, fragments can also be. e.g., at most 1000, at most 900, at most 800, at most 700, at most 600, at most 500, at most 450, at most 400, at most 350, at most 300, at most 250, at most 200, at most 150, at most 100, at most 50, at most 25, at most 10, or at most 5 amino acids in length. A fragment can further comprise, at either or both of its ends, one or more additional amino acids, for example, a sequence of amino acids from a different naturally occurring protein (e.g., an Fc or leucine zipper domain) or an artificial amino acid sequence (e.g., an artificial linker sequence).

[0410] Percent sequence identity is determined by conventional methods. See, for example, Altschul et al., Bull. Math. Bio. 48:603 (1986), and Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89: 10915 (1992). Briefly, two amino acid sequences are aligned to optimize the alignment scores using a gap opening penalty of 10. a gap extension penalty of 1, and the ■'BLOSUM62” scoring matrix of Henikoff and Henikoff (Id.). The sequence identity is then calculated as: ([Total number of identical matches] / [length of the longer sequence plus the number of gaps introduced into the longer sequence in order to align the two sequences])(100).

[0411] Additionally, there are many established algorithms available to align two amino acid sequences. For example, the “FASTA” similarity search algorithm of Pearson and Lipman is a suitable protein alignment method for examining the level of sequence identity or homology shared by an amino acid sequence of a peptide disclosed herein and the amino acid sequence of a peptide variant. The FASTA algorithm is described by Pearson and Lipman, Proc. Nat'l Acad. Sci. USA 85:2444 (1988), and by Pearson. e / / ?. Enzymol. 183:63 (1990). Briefly, FASTA first characterizes sequence similarity by identifying regions shared by the query sequence (e.g., SEQ ID NO: 9) and a test sequence that has either the highest density of identities (if the ktup variable is 1) or pairs of identities (if ktup=2), without considering conservative amino acid substitutions, insertions, or deletions. The ten regions with the highest densify of identities are then rescored by comparing the similarity of all paired amino acids using an amino acid substitution matrix, and the ends of the regions are ‘’trimmed” to include only those residues that contribute to the highest score. If there are several regions with scores greater than the “cutoff’ value (calculated by a predetermined formula based upon the length of the sequence and the ktup value), then the trimmed initial regions are examined to determine whether the regions can be joined to form an approximate alignment with gaps. Finally, the highest scoring regions of the two amino acid sequences are aligned using a modification of the Needleman-Wunsch-Sellers algorithm (Needleman and Wunsch, J. Mol. Biol. 48:444 (1970); Sellers, Siam J. Appl. Math. 26:787 (1974)), which allows for amino acid insertions and deletions. Illustrative parameters for FASTA analysis are: ktup=l, gap opening penalfy=10, gap extension penalfy=l, and substitution matrix=BLOSUM62. These parameters can be introduced into a FASTA program by modifying the scoring matrix file (“S MATRIX”), as explained in Appendix 2 of Pearson, Meth. EnzymolA83 63 (1990).

[0412] FASTA can also be used to determine the sequence identify of nucleic acid molecules using a ratio as disclosed above. For nucleotide sequence comparisons, the ktup value can range between one to six, preferably from three to six, most preferably three, with other parameters set as described above.

[0413] Some examples of common amino acids that are a “conservative amino acid substitution” are illustrated by a substitution among amino acids within each of the following groups: (1) glycine, alanine, valine, leucine, and isoleucine, (2) phenylalanine, tyrosine, and tryptophan, (3) serine and threonine. (4) aspartate and glutamate, (5) glutamine and asparagine, and (6) lysine, arginine and histidine. The BLOSUM62 table is an amino acid substitution matrix derived from about 2,000 local multiple alignments of protein sequence segments, representing highly conserved regions of more than 500 groups of related proteins (Henikoff and Henikoff, Proc. Nat’lAcad Sci. USA 89:10915 (1992)). Accordingly, the BLOSUM62 substitution frequencies can be used to define conservative amino acid substitutions that may be introduced into the amino acid sequences of the present invention. Although it is possible to design amino acid substitutions based solely upon chemical properties (as discussed above), the language “conser ative amino acid substitution” preferably refers to a substitution represented by a BLOSUM62 value of greater than -1. For example, an amino acid substitution is conservative if the substitution is characterized by a BLOSUM62 value of 0, 1, 2, or 3.According to this system, preferred conservative amino acid substitutions are characterized by a BLOSUM62 value of at least 1 (e.g., 1, 2 or 3), while more preferred conservative amino acid substitutions are characterized by a BLOSUM62 value of at least 2 (e.g., 2 or 3).

[0414] Determination of amino acid residues that are within regions or domains that are critical to maintaining structural integrity can be determined. Within these regions one can determine specific residues that can be more or less tolerant of change and maintain the overall tertiary structure of the molecule. Methods for analyzing sequence structure include, but are not limited to, alignment of multiple sequences with high amino acid or nucleotide identity’ and computer analysis using available software (e.g., the Insight II. RTM. viewer and homology modeling tools; MSI, San Diego, Calif.), secondary structure propensities, binary’ patterns, complementary7packing and buried polar interactions (Barton, G. J., Current Opin. Struct. Biol. 5:372-6 (1995) and Cordes, M H. et al., Current Opin. Struct. Biol. 6:3-10 (1996)). In general, when designing modifications to molecules or identifying specific fragments determination of structure can ty pically be accompanied by’ evaluating activity of modified molecules.

[0415] In another embodiment, the peptide comprises Compound 76, which is a chlorotoxin variant comprising the sequence of MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9), wherein the lysine residue is conjugated to a cyanine fluorescent label. The structure of Compound 76 is shown below7.

[0416] The structure of tozuleristide (Compound 76) is shown below:MCMPCFTTDHQMARRCDDCCGGRGRGKCYGPQCLCR (SEQ ID NO: 9).

[0417] The peptide can be further cross-linked by four disulfide bonds formed among the cysteine residues present in the sequence.TABLE 4 - Exemplary Compounds According to the Present Disclosure

[0418] In some aspects, the peptide is a variant of the native peptide of chlorotoxin but retains all eight cysteine residues of the native peptide, enabling cross-linking by up to four disulfide bonds. Conservation of cysteine residues helps to preserve the secondary structure and other features of the native chlorotoxin peptide because of the disulfide bonds that form between the cysteine residues. In some aspects, the chlorotoxin peptide variant retains all eight cysteineresidues of the native peptide and has at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 83%, 85%, 86%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the native chlorotoxin peptide.

[0419] In some aspects, the chlorotoxin peptide variant has eight cysteine residues positioned so that the distances between pairs of cysteines is the same as the distances between pairs of cysteines found in the native peptide, and the chlorotoxin peptide variant has at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 83%, 85%, 86%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the native chlorotoxin peptide.

[0420] In some aspects, the chlorotoxin peptide variant has eight cysteine residues positioned so that the distances between pairs of cysteines is functionally equivalent or functionally similar to the distances between pairs of cysteines found in the native peptide, and the chlorotoxin peptide variant has at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 83%, 85%, 86%, 89%, 90%, 92%, 93%, 94%, 95%, 96%. 97%. 98%. or 99% sequence identity with the native chlorotoxin peptide.

[0421] In some aspects, the chlorotoxin peptide variant has eight cysteine residues positioned so that the distances between pairs of cysteines allows for secondary' structure and isoelectric point of the native chlorotoxin peptide to be preserved, and the chlorotoxin peptide variant has at least 40%. 45%. 50%. 55%. 60%. 65%. 70%. 75%. 80%. 83%. 85%. 86%. 89%. 90%. 92%. 93%. 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the native chlorotoxin peptide.

[0422] In some aspects, the chlorotoxin peptide variant has eight cysteine residues positioned so that the distances between pairs of cysteines is sufficient to allow disulfide bonds to form, and the chlorotoxin peptide variant has at least 40%. 45%. 50%, 55%, 60%, 65%, 70%, 75%, 80%, 83%, 85%, 86%, 89%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the native chlorotoxin peptide.

[0423] In some aspects, one or more methionines of the chlorotoxin peptide variant are replaced with other amino acids. In some aspects, one or more methionines of the chlorotoxin peptide variant are replaced with other amino acids selected from glycine, alanine, isoleucine, threonine, valine, leucine, serine or a combination thereof.

[0424] In some embodiments, the chlorotoxin can be a chlorotoxin variant. Peptides are further described in PCT Patent Application Publication Numbers WO2006115633 andWO2011142858, which are incorporated in their entirety herein by reference.

[0425] In one embodiment, the peptide can have the following formula: H-Met-Cys-Met-Pro-Cys-Phe-Thr-Thr-Asp-His-Gln-Met-Ala-Arg-Xi-Cys-Asp-Asp-Cys-Cys-Gly-Gly-X2-Gly-Arg-Gly-X3-Cys-Tyr-Gly-Pro-Gln-Cys-Leu-Cys-Arg-OH (SEQ ID NO: 482) acetate salt (disulfide bonds, air oxidized), wherein Xi, X2, and X3 can each independently be any amino acid.

[0426] In one embodiment, the peptide can have the following formula: H-Met-Cys-Met-Pro-Cys-Phe-Thr-Thr-Asp-His-Gln-Met-Ala-Arg- Xi-Cys-Asp-Asp-Cys-Cys-Gly-Gly- X2-Gly-Arg-Gly- Xa-Cys-Tyr-Gly-Pro-Gln-Cys-Leu-Cys-Arg-OH (SEQ ID NO: 483) acetate salt (disulfide bonds, air oxidized), wherein Xi, X2, and X3 can each independently be Arg, Ala, or Lys.

[0427] In another embodiment, the all peptide can have the following formula: H-Met-Cys-Met-Pro-Cys-Phe-Thr-Thr-Asp-His-Gln-Met-Ala-Arg-Xi-Cys-Asp-Asp-Cys-Cys-Gly-Gly-Xz-Gly-Arg-Gly-Xs-Cys-Tyr-Gly-Pro-Gln-Cys-Leu-Cys-Arg-OH (SEQ ID NO: 484) acetate salt (disulfide bonds, air oxidized), wherein Xi, X2, and X3 can each independently be Arg or Ala.

[0428] In another embodiment, the all peptide can have the following formula: H-Met-Cys-Met-Pro-Cys-Phe-Thr-Thr-Asp-His-Gln-Met-Ala-Arg-Xi-Cys-Asp-Asp-Cys-Cys-Gly-Gly-X2-Gly-Arg-Gly-Lys-Cys-Tyr-Gly-Pro-Gln-Cys-Leu-Cys-Arg-OH (SEQ ID NO: 485) acetate salt (disulfide bonds, air oxidized), wherein Xi and X2 can each independently be Arg or Ala.

[0429] In another embodiment, the peptide can have the following formula: H-Met-Cys-Met-Pro-Cys-Phe-Thr-Thr-Asp-His-Gln-Met-Ala-Arg-Arg-Cys-Asp-Asp-Cys-Cys-Gly-Gly-Arg-Gly-Arg-Gly-Lys-Cys-Tyr-Gly-Pro-Gln-Cys-Leu-Cys-Arg-OH (SEQ ID NO: 9) acetate salt (disulfide bonds, air oxidized).

[0430] In another embodiment, the peptide can have the following formula: H-Met-Cys-Met-Pro-Cys-Phe-Thr-Thr-Asp-His-Gln-Met-Ala-Arg-Arg-Cys-Asp-Asp-Cys-Cys-Gly-Gly-Ala-Gly-Arg-Gly-Lys-Cys-Tyr-Gly-Pro-Gln-Cy s-Leu-Cys-Arg-OH (SEQ ID NO: 6) acetate salt (disulfide bonds, air oxidized).

[0431] In another embodiment, the peptide can have the following formula: H-Met-Cys-Met-Pro-Cys-Phe-Thr-Thr-Asp-His-Gln-Met-Ala-Arg-Ala-Cys-Asp-Asp-Cys-Cys-Gly-Gly-Arg-Gly-Arg-Gly-Lys-Cys-Tyr-Gly-Pro-Gln-Cys-Leu-Cys-Arg-OH (SEQ ID NO: 8) acetate salt (disulfide bonds, air oxidized).

[0432] In another embodiment, the peptide can have the following formula: H-Met-Cys-Met-Pro-Cys-Phe-Thr-Thr-Asp-His-Gln-Met-Ala-Arg-Ala-Cys-Asp-Asp-Cys-Cys-Gly-Gly-Ala-Gly-Arg-Gly-Lys-Cys-Tyr-Gly-Pro-Gln-Cys-Leu-Cys-Arg-OH (SEQ ID NO: 5) acetate salt (disulfide bonds, air oxidized).Linkers

[0433] In some aspects, the peptides of the present disclosure are directly conjugated to a detectable label, such as a dye. fluorescent moiety or the like such that no additional amino acids, carbohydrates, nucleic acids, polymers, organic chains, or the like are added to the peptideor peptide variant and / or the dye, fluorescent moiety or the like to comprise the peptide complexes described herein. In some other aspects, a linker is used to conjugate the peptide or peptide variant is not directly conjugated to a dye, fluorescent moiety or the like such that additional amino acids, carbohydrates, nucleic acids or the like are added to the peptide or peptide variant and / or the dye. fluorescent moiety or the like to comprise the peptide complexes described herein. A ‘linker” as used herein refers to at least one compound comprising two functional groups that are capable of reacting specifically with other moieties to form covalent or non-covalent linkages. Such moieties can include, but are not limited to, the side groups on naturally occurring amino acids or non-natural amino acids or peptides which contain such natural or non-natural amino acids. By way of example, a linker has a functional group reactive wi th a group on a first peptide, and another functional group which is reactive with a group on a second peptide, whereby forming a conjugate that includes the first peptide, the linker and the second peptide. Many procedures and linker molecules for attachment of various compounds to peptides are known. See, e.g., European Patent Application No. 188,256; U. S. Pat. Nos.4,671,958, 4,659,839, 4,414,148, 4,699,784; 4,680,338; and 4,569,789 which are incorporated by reference herein in their entirety. Linker moieties can include cleavable (e.g., pH sensitive or enzyme-labile linkers) or stable linkers.

[0434] The term “linkage,” as used herein refers to a bond or a chemical moiety formed from a chemical reaction between the functional group of a linker and another molecule. Such bonds can include, but are not limited to, covalent linkages and non-covalent bonds, while such chemical moieties include, but are not limited to, esters, carbonates, imines phosphate esters, hydrazones, acetals, orthoesters, peptide linkages, and oligonucleotide linkages. Hydrolytically stable linkages means that the linkages are substantially stable in water and do not react with water at neutral pH values, including but not limited to, under physiological conditions for an extended period of time, perhaps even indefinitely. Hydrolytically unstable or degradable linkages mean that the linkages are degradable in water or in aqueous solutions, including for example, blood. Enzymatically unstable or degradable linkages mean that the linkage is often degraded by one or more enzymes. By way of example, PEG and related polymers include degradable linkages in the polymer backbone or in the linker group between the polymer backbone and one or more of the terminal functional groups of the polymer molecule. Such degradable linkages can include, but are not limited to, ester linkages formed by the reaction of PEG carboxylic acids or activated PEG carboxylic acids with alcohol groups on a biologically active agent, wherein such ester groups generally hydrolyze under physiological conditions to release the biologically active agent. Other hydrolytically degradable linkages can include butare not limited to carbonate linkages; imine linkages resulted from reaction of an amine and an aldehyde; phosphate ester linkages formed by reacting an alcohol with a phosphate group; hydrazone linkages which are reaction product of a hydrazide and an aldehyde; acetal linkages that are the reaction product of an aldehyde and an alcohol; orthoester linkages that are the reaction product of a formate and an alcohol; peptide linkages formed by an amine group, including but not limited to, at an end of a polymer such as PEG, and a carboxyl group of a peptide; and oligonucleotide linkages formed by a phosphoramidite group, including but not limited to, at the end of a polymer, and a 5' hydroxyl group of an oligonucleotide.

[0435] The complexes for use in the method described herein can be conjugated by using any art-recognized method forming a complex including covalent, ionic, or hydrogen bonding of the ligand to the imaging agent, either directly or indirectly via a linking group such as a linker. The conjugate can typically be formed by covalent bonding of the ligand to the imaging agent through the formation of amide, ester or imino bonds between acid, aldehyde, hydroxy, amino, or hydrazo groups on the respective components of the complex or, for example, by the formation of disulfide bonds.

[0436] In addition, structural modifications of a linker portion of the complexes are contemplated herein. For example, a number of amino acid substitutions are often made to the linker portion of the conjugate, including but not limited to naturally occurring amino acids, as well as those available from conventional synthetic methods. In one aspect, beta, gamma, and longer chain amino acids are used in place of one or more alpha amino acids. In another aspect, the stereochemistry of the chiral centers found in such molecules is selected to form various mixture of optical purity of the entire molecule, or only of a subset of the chiral centers present. In another aspect, the length of the peptide chain included in the linker is shortened or lengthened, either by changing the number of amino acids included therein, or by including more or fewer beta, gamma, or longer chain amino acids. In another aspect, the selection of amino acid side chains in the peptide portion is made to increase or decrease the relative hydrophilicity of the linker portion specifically or of the overall molecule generally.

[0437] Similarly, the length and shape of other chemical fragments of the linkers described herein can often be modified. In some aspects, the linker includes an alkylene chain. The alkylene chain can often vary' in length, or can include branched groups, or can include a cyclic portion, which can be in line or spiro relative to the allylene chain. In another aspect, where the linker includes a beta thiol releasable fragment, it is appreciated that other intervening groups connecting the thiol end to the hydroxy or carbonate end are used in place of the ethylene bridge, such as but not limited to optionally substituted benzyl groups, where the hydroxy end isconnected at the benzyl carbon and the thiol end is connected through the ortho or para phenyl position, and vice versa.

[0438] Direct attachment can be achieved by covalent attachment of a peptide to another molecule. For example, the peptide is attached to a terminus of the amino acid sequence of a larger polypeptide or peptide molecule, or could be attached to a side chain, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, a non-natural amino acid residue, or glutamic acid residue. The attachment can be via an amide bond, an ester bond, an ether bond, a carbamate bond, a carbon-nitrogen bond, a triazole, a macrocycle, an oxime bond, a hydrazone bond, a carbon-carbon single double or triple bond, a disulfide bond, or a thioether bond. In some embodiments, similar regions of the disclosed peptide(s) itself (such as a terminus of the amino acid sequence, an amino acid side chain, such as the side chain of a lysine, serine, threonine, cysteine, ty rosine, aspartic acid, a non-natural amino acid residue, or glutamic acid residue, via an amide bond, an ester bond, an ether bond, a carbamate bond, a carbon-nitrogen bond, a triazole, a macrocycle, an oxime bond, a hydrazone bond, a carbon-carbon single double or triple bond, a disulfide bond, or a thioether bond, or linker as described herein) may be used to link other molecules.

[0439] Attachment via a linker can involve incorporation of a linker moiety between the other molecule and the peptide. The peptide and the other molecule can both be covalently attached to the linker. The linker can be cleavable, stable, self-immolating, hydrophilic, or hydrophobic. The linker can have at least two functional groups, one bonded to the other molecule, one bonded to the peptide, and a linking portion between the two functional groups. The use of a cleavable linker can permit release of the conjugated moiety (e.g., a detectable agent or a therapeutic agent) from the peptide, e.g., after targeting to a tissue of interest. The cleavable linker can comprise a cleavage site for matrix metalloproteinases, thrombin, cathepsins, or betaglucuronidase. In other aspects, the linker can be a hydrolytically labile linker. A hydrolytically labile linker, (amongst other cleavable linkers described herein) can be advantageous in terms of releasing a fluorophore molecule or other detectable or therapeutic agents from the peptide. For example, an agent (e g., a detectable agent or a therapeutic agent) in a conjugate form with the peptide may not be active, but upon release from the conjugate after targeting to the cartilage, the agent can be active. In some cases, the linker can be enzyme cleavable, e.g., a valinecitrulline linker. Alternatively or in combination, the linker can be cleavable by other mechanisms, such as via pH, reduction, or hydrolysis. Other cleavable linkers can include an ester bond using standard l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)-, dicylcohexylcarbodiimide (DCC)-, thionyl chloride-, or phosphorous chloride-basedbioconjugation chemistries. These linkers can be cleaved by esterases, MMP, cathepsin B, a protease, or thrombin. In still other aspects, the peptide can be linked to the detectable agent via a stable linker.

[0440] Non-limiting examples of the functional groups for attachment can include functional groups capable of forming, for example, an amide bond, an ester bond, an ether bond, a carbonate bond, a carbamate bond, or a thioether bond. Non-limiting examples of functional groups capable of forming such bonds can include amino groups; carboxyl groups; hydroxyl groups: aldehyde groups; azide groups; alkyne and alkene groups; ketones; hydrazides; acid halides such as acid fluorides, chlorides, bromides, and iodides; acid anhydrides, including symmetrical, mixed, and cyclic anhydrides; carbonates; carbonyl functionalities bonded to leaving groups such as cyano, succinimidyl, and V-hydroxysuccinimidyl; hydroxyl groups; sulfhydryl groups; and molecules possessing, for example, alkyl, alkenyl, alkynyl, allylic, or benzylic leaving groups, such as halides, mesylates, tosylates, tritiates, epoxides, phosphate esters, sulfate esters, and besylates.

[0441] Non-limiting examples of the linking portion can include alkylene, alkenylene, alkynylene, polyether, such as polyethylene glycol (PEG), hydroxy carboxylic acids, polyester, polyamide, polyamino acids, polypeptides, cleavable peptides, valine-citrulline, aminobenzylcarbamates. D-amino acids, and poly amine, any of which being unsubstituted or substituted with any number of substituents, such as halogens, hydroxyl groups, sulfhydryl groups, amino groups, nitro groups, nitroso groups, cyano groups, azido groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, carboxaldehyde groups, imine groups, alkyl groups, halo-alkyl groups, alkenyl groups, halo-alkenyl groups, alkynyl groups, halo-alkynyl groups, alkoxy groups, aryl groups, aryloxy groups, aralkyl groups, arylalkoxy groups, heterocyclyl groups, acyl groups, acyloxy groups, carbamate groups, amide groups, urethane groups, epoxides, and ester gr...

Claims

CLAIMSWhat is claimed is:

1. A method of detecting a tissue or cell and a nene tissue or cell in a subject in need thereof, the method comprising administering to a subject a tumor homing agent and a nerve homing agent.

2. The method of claim 1. wherein the detecting comprises fluorescence imaging.

3. The method of claim 1 or claim 2, wherein the tumor homing agent is fluorescent, the nerve homing agent is fluorescent, or both.

4. The method of any one of claims 1-3, wherein the nerve homing agent and the tumor homing agent have a minimum spectral gap in emission wavelength of at least 5 nm to at least 800 nm.

5. The method of claim 4. wherein the minimum spectral gap in emission wavelength is at least 5 nm, at least 10 nm, at least 20 nm, at least 30 nm, at least 40 nm, at least 50 nm, at least 60 nm, at least 70 nm, at least 80 nm, at least 90 nm, at least 100 nm, at least 110 nm, at least 120 nm, at least 130 nm, at least 140 nm, at least 150 nm, at least 160 nm, at least 170 nm, at least 180 nm. at least 190 nm, or at least 200 nm.

6. The method of any one of claims 1-5, wherein an emission wavelength spectrum of the nerve homing agent overlaps an excitation wavelength spectrum of the tumor homing agent by at least 0.5%, at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, or at least 40%.

7. The method of any one of claims 1-6, wherein the nerve homing agent and tumor homing agent have a spectral gap in peak excitation wavelength of at least 5 nm to at least 800 nm.

8. The method of claim 7, wherein the spectral gap in peak excitation wavelength of the nerve homing agent relative to the tumor homing agent is at least 5 nm, at least 10 nm, at least 20 nm, at least 30 nm, at least 40 nm, at least 50 nm, at least 60 nm, at least 70 nm, at least 80 nm, at least 90 nm, at least 100 nm, at least 110 nm, at least 120 nm, at least 130 nm, at least140 nm, at least 150 nm, at least 160 nm, at least 170 nm, at least 180 nm, at least 190 nm, or at least 200 nm.

9. The method of any one of claims 1-8, wherein an excitation wavelength spectrum of the nerve homing agent does not overlap with an emission wavelength spectrum of the tumor homing agent.

10. The method of any one of claims 1-8, wherein an excitation wavelength spectrum of the nerve homing agent completely or partially overlaps an emission wavelength spectrum of the tumor homing agent.

11. The method of any one of claims 1-8, wherein an emission wavelength spectrum of the nerve homing agent does not overlap an excitation wavelength spectrum of the tumor homing agent.

12. The method of any one of claims 1-8, wherein an emission wavelength spectrum of the nerve homing agent completely or partially overlaps an excitation wavelength spectrum of the tumor homing agent.

13. The method of any one of claims 1-12, wherein the tumor homing agent has a peak emission between 600 to 900 nm.

14. The method of any one of claims 1-13. wherein the tumor homing agent is excited, imaged, detected, or any combination thereof, at 715 to 800 nm excitation.

15. The method of any one of claims 1-14, wherein the tumor homing agent is detected at 815 to 875 nm emission.

16. The method of any one of claims 1-15, wherein the tumor homing agent detects a physiologic structure or cell between at least 10 pm and 25 pm in diameter.

17. The method of any one of claims 1-16. wherein the nerve homing agent has a peak emission between 600-775 nm inclusive.

18. The method of any one of claims 1-17, further comprising performing a fluorescence guided surgery (FGS) procedure on the subject in need thereof.

19. The method of claim 18, wherein the fluorescence guided surgery (FGS) is performed with the assistance of an imaging system.

20. The method of claim 19, wherein the imaging system is selected from the group consisting of: surgical microscope, confocal microscope, fluorescence scope, exoscope,endoscope, ophthalmoscope, retinal camera system, optical coherence tomography (OCT) system, surgical robot, and Quest Spectrum imaging system, Fluobeam devices, Lab Flare imaging system, or Solaris imaging system, or other imaging system equipped with two-color imaging capabilities.

21. The method of any one of claims 1-20. wherein the nerve homing agent is retained by, or accumulates in and / or binds to a nerve tissue or nerve cell.

22. The method of any one of claims 1-21, further comprising detecting the presence or absence of the nen e homing agent in a tissue or cell, wherein the presence of the nerve homing agent in the tissue or cell indicates the presence of nerve tissue or a nerve cell.

23. The method of any one of claims 1-22, w herein the tumor homing agent is retained by, or accumulates in and / or binds to cancerous tissue or a cancer cell, or a vascular lesion.

24. The method of any one of claims 1-23, further comprising detecting the presence or absence of the tumor homing agent in a tissue or cell, w herein the presence of the tumor homing agent in the tissue or cell indicates the presence of a cancerous tissue or a cancer cell, or a vascular lesion.

25. The method of any one of claims 1-24, further comprising treating a cancer or a vascular lesion in the subject in need thereof.

26. The method of claim 25, wherein the treating further comprises a surgery for surgically removing a cancerous tumor, a cancerous tissue, or a cancerous cell, or vascular lesion, from the subject in need thereof by performing a surgery.

27. The method of claim 26, wherein surgically removing the cancerous tumor, the cancerous tissue, or the cancerous cell, or vascular lesion, from the subject in need thereof spares the nene tissue or nerve cell, avoids damaging the nerve tissue or nerve cell, or does not damage a nerve tissue or nerve cell in the subject in need thereof.

28. The method of claim 26 or 27, wherein surgically removing the cancerous tumor, the cancerous tissue, or the cancerous cell, or vascular lesion, from the subject in need thereof shows sparing the nerve tissue or nerve cell, avoiding damaging the nerve tissue or nerve cell, or not damaging a nerve tissue or nerve cell ex vivo in pathology7samples obtained from the subject in need thereof or in situ in the patient in need thereof.

29. The method of any one of claims 26-28, wherein the surgery is fluorescence guided surgery (FGS).

30. The method of any one of claims 25-29, wherein the cancer is glioma, astrocytoma, medulloblastoma, choroids plexus carcinoma, ependymoma, neuroblastoma, vestibular schwannoma, carcinoma, meningioma, head and neck cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, breast cancer, intestinal cancer, pancreatic cancer, liver cancer, kidney cancer, sarcoma, osteosarcoma, rhabdomyosarcoma, Ewing’s sarcoma, gastrointestinal stromal tumors, melanoma, ovarian cancer, cervical cancer, lymphoma, thyroid cancer, anal cancer, colorectal cancer, endometrial cancer, laryngeal cancer, multiple myeloma, prostate cancer, retinoblastoma, gastric cancer, testicular cancer, Wilm’s tumor, or any combination thereof, or where the vascular lesion is a cavernous angioma, cavernous hemangioma, or cerebral cavernous malformation (CCM), or any combination thereof.

31. The method of any one of claims 25-30, wherein the cancer is a head and neck cancer, a brain cancer, a brain tumor, breast cancer, melanoma, sarcoma, basal cell carcinoma, squamous cell carcinoma, lung cancer, colorectal cancer, prostate cancer, bladder cancer, or any combination thereof.

32. The method of claim 31, wherein the head and neck cancer is a head and neck squamous cell carcinoma.

33. The method of claim 31 or 32, wherein the breast cancer is invasive ductal carcinoma, ductal carcinoma in situ breast cancer, invasive lobular carcinoma, lobular carcinoma in situ breast cancer, triple negative breast cancer, or any combination thereof.

34. The method of any one of claims 1-33, wherein the tumor homing agent, the nerve homing agent, or both, are formulated for intravenous administration, intravenous bolus administration, intravenous infusion administration, intramuscular administration, subcutaneous administration, intraperitoneal administration, oral administration, sublingual administration, topical administration, transdermal administration using electroporation, intralesional administration, or a combination thereof.

35. The method of any one of claims 1-34, wherein the tumor homing agent and the nerve homing agent are co-administered or are sequentially administered.

36. The method of any one of claims 1-35, wherein the tumor homing agent is administered prior to the nerve homing agent, or wherein the nen e homing agent isadministered prior to the tumor homing agent, or wherein the tumor homing agent and the nerve homing agent are administered at the same time.

37. The method of any one of claims 1-36, wherein the tumor homing agent is administered 1 to 24 hours before or 1 to 3 days before a surgery and the nerve homing agent is administered 10 to 120 minutes before or 2 to 48 hours before a surgery.

38. The method of any one of claims 1-37. wherein the tumor homing agent comprises a polypeptide comprising:(a) a sequence of any one of SEQ ID NO: 482 - SEQ ID NO: 485, or a fragment thereof,(b) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512, or a fragment thereof, or(c) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to SEQ ID NO:

9. or a fragment thereof.

39. The method of claim 38, wherein the polypeptide comprises SEQ ID NO: 9, or a fragment thereof.

40. The method of claim 38 or claim 39, wherein the fragment of the polypeptide has a length of at least 25 residues.

41. The method of any one of claims 38-40, wherein each amino acid of the polypeptide is independently selected as an L- or D-enantiomer.

42. The method of any one of claims 38-41, wherein the polypeptide contains no lysine residues.

43. The method of any one of claims 38-42, wherein the polypeptide contains a single lysine residue.

44. The method of claim 43, wherein the single lysine residue is located at a position corresponding to K-27 of native chlorotoxin, K-23 of native chlorotoxin, or K-15 of native chlorotoxin.

45. The method of any one of claims 38-44, wherein one, two, or three methionine residues of the polypeptide are replaced with other amino acids.

46. The method of any one of claims 38-45, wherein the N-terminus of the polypeptide is blocked by acetylation or cyclization47. The method of any one of claims 38-46, wherein the polypeptide compnses at least 4, at least 5, or at least 6 disulfide bonds.

48. The method of any one of claims 38-47, wherein the polypeptide has a length of no less than 21 amino acid residues, no less than 22 amino acid residues, no less than 23 amino acid residues, no less than 24 amino acid residues, no less than 25 amino acid residues, no less than 26 amino acid residues, no less than 27 amino acid residues, no less than 28 amino acid residues, no less than 29 amino acid residues, no less than 30 amino acid residues, no less than 31 amino acid residues, no less than 32 amino acid residues, no less than 33 amino acid residues, no less than 34 amino acid residues, no less than 5 amino acid residues, no less than 36 amino acid residues, no less than 37 amino acid residues, no less than 38 amino acid residues, no less than 39 amino acid residues, or no less than 40 amino acid residues.

49. The method of any one of claims 38-48, wherein the polypeptide comprises an isoelectric point at least 8.0, at least 8.5, or at least 9.0.

50. The method of any one of claims 38-49, wherein the polypeptide is conjugated to an agent.

51. The method of claim 50, wherein the agent is a detectable agent or a first therapeutic agent.

52. The method of claim 50 or claim 51, wherein the polypeptide is conjugated to the agent via a cleavable linker or a stable linker.

53. The method of any one of claims 50-52, wherein the polypeptide comprises:(a) a single lysine residue and the agent is conjugated to the polypeptide at the single lysine residue, or(b) no lysine residues and the agent is conjugated to the polypeptide at the N- terminus of the polypeptide.

54. The method of any one of claims 1-53, wherein the tumor homing agent comprises the structure of Formula (I), or a pharmaceutically acceptable salt thereof:R1, R2, R3, R4, R5, R6, R7, R8, R15, and R16are each independently selected from hydrogen. Ci-Ce alkyl. Ci-Cs alkylene-COOH, sulfonate, Ci-Ce alkylene-sulfonate, -COOH, -SO2-NH2, or Ci-Ce alkoxy;R9is hydrogen, sulfonate, amine, or COOH;L1is C3-C6 alkylene;L² is C1-C10 alkylene;L3is a bond, -O-, -NR10-, -NR10-Ci-C6alkylene- -O-NR10-, - R10-CI-C6alkylene- (O-Ci-Ce alkylene)n-, -NR10-L4-, - R10-CI-C6 alkylene-NR11- (C (= O) -Ci-Ce alkylene-O-) -, or -NR10-CI-C6 alky lene— NR10-Ci-Ce. alkylene— NR!0-CI-C6alkylene-;L4is a bond, -heterocyclyl ---. or -heterocyclyl-Ci-Cs alkylene-;R10is hydrogen or Ci-Ce alkyl;R11is hydrogen or Ci-Cs alkyl;R12and R13are independently selected from hydrogen, Ci-Ce alkyl, or R12and R13are joined together along with the other atoms to which they’ are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;R14is hydrogen or C1-C.5 alkylene, -(L’j-aryl, -(L )-aryl-R21, -(L5)-heteroaiyl, -(L5)-heteroaryl-R2!, -NR17R18, R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring;L5is a bond, C1-C10 alkylene, O, -NR10-;R17and R18are each independently hydrogen or aryl;R19and R20are independently' selected from hydrogen, Ci-Ce alkyl. R14and R19are joined together along with the other atoms to which they are attached to form a 5-membered or 6-membered carbocyclic or heterocyclic ring, or R14and R20are joined together along with the other atoms to which they are attached to form a 5 -membered or 6-membered carbocyclic or heterocyclic ring;R21is hydrogen, sulfonate, or -COOH;n is 0, 1, 2, or 3;m is 0, 1, 2, or 3;p is 0, 1, 2, or 3;q is 0. 1, 2, or 3; andA4is a polypeptide.

55. The method of claim 54, wherein R9is sulfonate.

56. The method of claim 54 or claim 55, wherein A4has at least 90% sequence identity with:(a) a sequence of any one of SEQ ID NO: 482 - SEQ ID NO: 485, or a fragment thereof.(b) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with any one of SEQ ID NO: 1 - SEQ ID NO: 481 or SEQ ID NO: 486 - SEQ ID NO: 512, or a fragment thereof, or(c) at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to SEQ ID NO:

9. or a fragment thereof.

57. The method of any one of claims 54-56, wherein A4is SEQ ID NO: 9 or a fragment thereof.

58. The method of any one of claims 54-57, wherein A4comprises at least 25 amino acid residues.

59. The method of any one of claims 54-58, wherein:R3, R4, R5, R6of the Formula (I), are each independently methyl;R1, R2, R7, R8, R15, and R16are each independently hydrogen;R10is hydrogen; R12, R13, R14, R19, and R20are each independently hydrogen;L1is butylene; andL2is pentylene.

60. The method of any one of claims 54-59, wherein L3is attached to A4at a lysine amino acid residue of the polypeptide.

61. The method of any one of claims 54-60, wherein Formula (I) is conjugated to polyethylene glycol (PEG), hydroxyethyl starch, polyvinyl alcohol, a water soluble polymer, a zwitterionic water soluble polymer, a water soluble poly(amino acid), an albumin, or a fatty7acid.

62. The method of any one of claims 54-61, wherein Formula (II) further comprises a second therapeutic agent attached to A4, wherein the second therapeutic agent is selected from a radioisotope, toxin, cytotoxic agent, enzyme, sensitizing drug, nucleic acid, interfering RNA, antibody, anti-angiogenic agent, cisplatin, anti-metabolite, mitotic inhibitor, grow th factor inhibitor, paclitaxel, temozolomide, topotecan, fluorouracil, vincristine, vinblastine, procarbazine, dacarbazine. altretamine, methotrexate, mercaptopurine, thioguanine, fludarabine phosphate, cladribine, pentostatin, cytarabine, azacitidine, etoposide, teniposide, irinotecan, docetaxel, doxorubicin, daunorubicin, dactinomycin, idarubicin, plicamycin, mitomycin, bleomycin, tamoxifen, flutamide, leuprolide, goserelin, aminogluthimide, anastrozole, amsacrine, asparaginase, mitoxantrone, mitotane, amifostine or a combination thereof.

63. The method of any one of claims 1-62. wherein the tumor homing agent comprises the structure of any one of Formulas (IX), (X), (XI), (XII), (XIII), (XIV), (XV), or (XVI), wherein A4is the polypeptide:

64. The method of any one of claims 48-63, wherein the detectable agent comprises a dye, a fluorophore, a fluorescent biotin compound, a luminescent compound, a chemiluminescent compound, a radioisotope, nanoparticle, a paramagnetic metal ion, a tumor homing agent, a nerve homing agent, and a combination thereof.

65. The method of any one of claims 48-64, wherein the detectable agent has a peak absorbance of 200 nm to 900 nm.

66. The method of any one of claims 48-64, wherein the detectable agent has a peak emission of 300 nm to 1000 nm.

67. The method of any one of claims 48-66, wherein the detectable agent has a peak emission of 400 nm to 600 nm, 450 nm to 600 nm, 500 nm to 600 nm, 550 nm to 600 nm, 400 nm to 550 nm, 400 nm to 500 nm, 400 nm to 450 nm, or 450 nm to 550 nm.

68. The method of any one of claims 48-67, wherein the detectable agent has a peak emission between the 600 to 900 nm or 600 to 775 nm wavelength.

69. The method of any one of claims 64-68, wherein:the dye comprises DyLight-680, DyLight-750, VivoTag-750, DyLight-800, IRDye-800, VivoTag-680, Cy5, Cy5.5, Cy7, Cy7.5, an indocyanine green (ICG), near infrared dyes, acridine orange or yellow, 7-actinomycin D, 8-anilinonaphthalene-l -sulfonic acid, ATTO dye and any derivative thereof, auramine-rhodamine stain and any derivative thereof, benzanthrone, bimane, 9-10-bis(phenylethynyl)anthracene, 5,12 - bis(phenylethynyl)naththacene, bisbenzimide, brainbow, calcein, carboxyfluorescein and any derivative thereof, 1 -chi oro-9, 10-bis(phenylethynyl)anthracene and any derivative thereof, DAPI, DiOC6, DyLight Fluors and any derivative thereof, epicocconone, ethidium bromide, FlAsH-EDT2, Fluo dye and any derivative thereof, FluoProbe and any derivative thereof, Fluorescein and any derivative thereof. Fura and any derivative thereof, GelGreen and any derivative thereof, GelRed and any derivative thereof, fluorescent proteins and any derivative thereof, m isoform proteins and any derivative thereof, heptamethine dye and any derivative thereof, hoeschst stain, iminocoumarin, indiarythrone, indo-1 and any derivative thereof, laurdan, lucifer yellow and any derivative thereof, luciferin and any derivative thereof, luciferase and any derivative thereof, mercocyanine and any derivative thereof, methylene blue and any derivative thereof, nile dyes and any derivative thereof, OS680, OS750, perylene, phloxine, phyco dye and any derivative thereof, propidium iodide, pyranine, rhodamine and any derivative thereof, ribogreen, RoGFP, rubrene, stilbene and any derivative thereof, sulforhodamine and any derivative thereof. SYBR and any derivative thereof, synapto-pHluorin, tetraphenyl butadiene, tetrasodium tris, Titan Yellow, topotecan, TSQ, umbelliferone, violanthrone, yellow fluroescent protein, YOYO-1 and ZW800, fluorescein and fluorescein dyes, carbocyanine, merocyanine, styryl dyes, oxonol dyes, phycoerythrin, erythrythrosinein, rhodamine dyes, coumarin and coumarin dyes, Oregon Green Dyes, Texas Red, Texas Red-X, SPECTRUM RED, SPECTRUM GREEN, cyanine dyes, ALEXA FLUOR dyes and any derivative thereof, BODIPY dyes, IRDyes, or any combination thereof;optionally wherein the m isoform proteins and any derivative thereof comprises mCherry;optionally wherein the fluorescein and fluorescein dyes comprise fluorescein isothiocyanate or FITC, naphthofluorescein, 4', 5' -dichloro-2',7' -dimethoxyfluorescein, or 6-carboxyfluorescein or FAM;optionally wherein the rhodamine dyes comprise carboxytetramethyl-rhodamine or TAMRA, carboxyrhodamine 6G, carboxy-X-rhodamine (ROX), lissamine rhodamine B, rhodamine 6G, rhodamine Green, rhodamine Red, or tetramethylrhodamine (TMR;optionally wherein the coumarin and coumarin dyes comprise methoxy coumarin, dialkylaminocoumarin, hydroxy coumarin, or aminomethylcoumarin (AMCA);optionally wherein the Oregon Green Dyes comprise Oregon Green 488, Oregon Green 500, or Oregon Green 514;optionally wherein the SPECTRUM GREEN comprises a cyanine dye comprising CY-3. Cy-5, CY-3.5, or CY-5.5;optionally wherein the ALEXA FLUOR dyes comprise ALEXA FLUOR 350, ALEXA FLUOR 488, ALEXA FLUOR 532, ALEXA FLUOR 546, ALEXA FLUOR 568, ALEXA FLUOR 594. ALEXA FLUOR 633, ALEXA FLUOR 660, or ALEXA FLUOR 680;optionally wherein the BODIPY dyes comprise BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650, or BODIPY 650 / 665;optionally wherein the IR Dyes comprise IRD40, IRD 700, or IRD 800;optionally wherein the radioisotope comprises iodine-131, iodine-125, bismuth-212, bismuth-213, lutetium-177, rhenium-186, rhenium-188, yttrium-90, astatine-211, phosphorus-32 and / or samarium-153, or an isotope having one or more atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature, orany combination thereof.

70. The method of claim 69, wherein the near infrared dye comprises a cyanine dyes.

71. The method of claim 69 or 70, wherein the dye is selected from indocyanine green (ICG), Cy5, Cy5.5, Cy7, Cy7.5, and IRDye800cw.

72. The method of claim 71, wherein the indocyanine green (ICG) has a peak absorbance of 750 nm to 770 nm, 755 nm to 775 nm, or 768 nm.

73. The method of claim 71 or 72, wherein the indocyanine green (ICG) has a peak emission of 790 nm to 820 nm, 805 nm to 815 nm, or 807 nm.

74. The method of any one of claims 68-73, wherein the isotope having one or more atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature comprises hydrogen, carbon, fluorine, phosphorous, copper, gallium, yttrium, technetium, indium, iodine, rhenium, thallium, bismuth, astatine, samarium, and lutetium, or any combination thereof.

75. The method of claim 74, wherein the lutetium comprises3H,2H,13C,14C,18F,32P,35S,64Cu,67Ga,90Y,99mTc,111In,125I,123I,131I,135I,186Re,187Re,201Tl,212Bi,211At,153Sm, or177Lu.

76. The method of any one of claims 1-75, wherein the tumor homing agent comprises a therapeutic agent selected from a radioisotope, nanoparticle, toxin, enzyme, sensitizing drug, radiosensitizer, photosensitizer, nucleic acid, interfering RNA, antibody, antibody fragment, aptamer, anti-angiogenic agent, anti -metabolite, mitotic inhibitor, grow th factor inhibitor, or any combination thereof.

77. The method of any one of claims 1-76, wherein the nerve homing agent is a molecule of Formula (DI):R3 is hydrogen or R2and R3 together form a fused ring, creating a core of FormulaR4 and Rs, together with the nitrogen atom to which they are bound, form a ring selected from the group of:or, when the compound is of Formula (DII), R4 and Rs may be independently selected from Ci-Ce alkyl, with the proviso that, when R4 is ethyl, Rs is not ethyl; Re is hydrogen;or, when R2 and R3 together form a fused ring to create a core of Formula (Dill), Rs and Re may also, together with the nitrogen atom to which Rs is bound, form afused ring, creating a core of Formula (Dill):with the proviso that, when R4 and Rs, together with the nitrogen atom to which theyare bound, form the ring, and R3 is H, Ri and R2, together with the nitrogen atom to which they are bound, may form a pyrrolidinyl ring;Xi in each instance is independently selected from Ci-Ce straight or branched alkyl, Ci-Ce straight or branched alkenyl, Ci-Ce straight or branched alkynyl, and - Si(Ci-C4alkyl)3;n is an integer selected from the group of 1 and 2;nl is an integer independently selected in each instance from the group of 1, 2, 3. and 4;n2 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;n3 is an integer independently selected in each instance from the group of 1, 2, 3. 4, 5, 6, 7, 8, 9, and 10;n4 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; andwith the proviso that the sum of n2 + n2 is not greater than 10;with the proviso that the sum of n2 + n3 is not greater than 10;with the proviso that the sum of n2 + n4 is not greater than 10;with the proviso that the sum of n3 + n4 is not greater than 10;with the proviso that, when R4 and Rs, together with the nitrogen atom to which theyare bound, form the ring, Ri and R2 are not both methyl, Ri and R2 arenot both ethyl, Ri and R2 are not both n-propyl, Ri and R2 are not both n-butyl, and Ri and R2 are not both n-pentyl;with the proviso that, when the compound is of Formula (DI), when Ri is methyl, R4 is not methyl; andwith the proviso that, when the compound is of Formula (DI), when Ri is ethyl, R4 is not ethyl.

78. The method of any one of claims 1-77, wherein the nerve homing agent is a molecule of Formula (DIV):wherein:R1is C 1-6 alkyl.

79. The method of any one of claims 1-78, wherein the nerve homing agent homing agent is selected from:

80. The method of any one of claims 1-79, wherein the nerve homing agent homing agent is detected at a 500-725 nm excitation.

81. The method of any one of claims 1-80, wherein the nerve homing agent homing agent is detected at a 600-800 nm emission.

82. The method of any one of claims 1-81. wherein the nerve homing agent homing agent is:

83. The method of any one of claims 1-82, wherein the nerve homing agent homing agent is detected at a 655 nm peak excitation.

84. The method of any one of claims 1-83, wherein the nerve homing agent homing agent is detected at a 678 nm peak emission.

85. The method of any one of claims 1-76, wherein the nerve homing agent is a molecule of Formula (El):Xi in each instance is independently selected from the group of Ci-Ce straight or branched alky l, C2-C6 straight or branched alkenyl, Ci-Ce straight or branched alkynyl, and -Si(Ci-C4 alkyl)3;nl is an integer independently selected in each instance from the group of 1, 2, 3. and 4;n2 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;n3 is an integer independently selected in each instance from the group of 1, 2, 3. 4, 5, 6, 7, 8, 9, and 10;n4 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; andn5 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5. 6, 7, 8. 9, and 10;with the proviso that the sum of n2 + n4 is not greater than 10;with the proviso that the sum of n3 + n5 is not greater than 10;with the proviso that the sum of n2 + n5 is not greater than 10; andwith the proviso that the sum of n3 + n4 is not greater than 10.

86. The method of any one of claims 1-76 or 85, wherein the nerve homing agent homing agent is selected from:

87. The method of any one of claims 1-76, 85, or 86, wherein the nerve homing agent homing agent is detected at a 450 to 700 nm excitation.

88. The method of any one of claims 1-76 or 85-87, wherein the nerve homing agent homing agent is detected at a 600 to 800 nm emission.

89. The method of any one of claims 1-76 or 85-88, wherein the nerve homing agent homing agent is detected at a 625 to 640 nm excitation.

90. The method of any one of claims 1-76 or 85-89, wherein the nerve homing agent homing agent is detected at a 655 to 661 nm emission.

91. The method of any one of claims 1-76, wherein the nerve homing agent is a molecule of Formula (FI):wherein:one of R1and R2is hydrogen;the other of R1and R2is selected from the group of:R3and R4are independently selected from the group of H, -CN. -O-Ci-Ce alkyl, — NH2, NH(C1-C6alkyl), — N(C1-C6alkyl)2, — CH=O, and — SO2(Ci-C6 alkyl); with the proviso that not more than one of R3 and R4 is hydrogen.

92. The method of any one of claims 1-76 or 91, wherein the nerve homing agent homing agent is selected from:

93. The method of any one of claims 1-76. wherein the nerve homing agent is a molecule of Formula (CI):wherein:X is selected from the group of a single bond and a double bond;Y is selected from the group of a single bond and a double bond;with the proviso that at least one of X and Y is a single bond;Ri is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R2 is a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;nl in each instance is an integer independently selected from the group of 1, 2, and 3; n2 in each instance is an integer independently selected from the group of 0, 1, and 2; or Ri and R2 together form a heterocyclic ring selected from the group of azetidinyl, pyrrolidinyl, and piperidinyl, wherein the heterocyclic ring formed by Ri and R2 is substituted by 1 or 2 substituents selected from the group of methoxy and ethoxy;or Ri and R2 together with the nitrogen atom to which they are bound form a spirocyclic heterocycle of the formula:n3 in each instance is an integer independently selected from the group of 0, 1, 2. and 3; n4 in each instance is an integer independently selected from the group of 0, 1, 2. and 3; with the proviso that the sum of n3 and n4 is not less than 2 and not greater than 4; n5 in each instance is an integer independently selected from the group of 0, 1, 2, and 3; n6 in each instance is an integer independently selected from the group of 0, 1, 2, and 3; with the proviso that the sum of n5 and n6 is not less than 2 and not greater than 4; R3 is selected from the group of Ci-Ce alkyl and a moiety of the formula -(CH2)ni-O-(CH2)n2-CH3;R4 is Ci-Ce alkyl when R3 is Ci-Ce alkyl; and R4 is -(CH2)ni-O-(CH2)n2-CH3 when R3 is -(CH2)nl-O-(CH2)n2-CH3;with the proviso that the integers of nl and n2 in the R4 -(CH2)ni-O-(CH2)n2-CH3 moiety may be the same or different from the integers of nl and n2 in the R3 -(CH2)ni-O-(CH2)n2-CH3moiety;Rs is H; or Rs is an oxygen atom that joins with R4 to form a 2,3,4,5a,6a,lla-hexahydro-[l,4]oxazino[2,3-b]phenoxazine compound of Formula (CII):Re is a ring heteroatom selected from the group of O and S;or a pharmaceutically acceptable salt, co-crystal, ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

94. The method of any one of claims 1-76 or 93, wherein the nerve homing agent homing agent is selected from:pharmaceutically acceptable salt, co-crystal. ester, solvate, hydrate, isomer (including optical isomers, racemates, or other mixtures thereof), tautomer, isotope, polymorph, or pharmaceutically acceptable prodrug thereof.

95. The method of any one of claims 1-76, wherein the nerve homing agent is a molecule of Formula (GI):R1and R2are each independently selected from hydrogen and C1-C4 alkyl; R3aand R3bare independently selected from hydrogen and C1-C4 alkyl, with the proviso that at least one of R3aand R3bis not hydrogen; or R3aand R3btogether with the nitrogen atom to which they are bound form a 5- or 6-membered nitrogen-containing ring;or R3bis selected from hydrogen and C1-C4 alkyl, and R3atogether with the nitrogen to which it is bound and R4forms a fused 5-membered or 6-membered ring, the 5-membered or 6-membered ring having one nitrogen heteroatom, two nitrogen heteroatoms, or one nitrogen heteroatom and one oxygen heteroatom, wherein each nitrogen heteroatom is optionally substituted by a C1-C4 alkyl substituent; or R3a, the nitrogen to which it is bound, and R4form a fused 6-membered, nitrogencontaining ring and R3b, the nitrogen to which it is bound, and R2together form a fused 6-membered, nitrogen-containing ring;R4and R7are each independently selected from hydrogen, halogen, or C1-C4 alkyl;R5and R6are each independently selected from hydrogen or C1-C4 alkyl;R8aand R8bare independently selected from hydrogen and C1-C4 alkyl, with the proviso that at least one of R8aand R8bis not hydrogen; or R8aand R8btogether with the nitrogen to which they are bound form a 5- or 6-membered nitrogencontaining ring optionally substituted by 1, 2, 3, or 4 C1-C4 alkyl substituents; or R8bis selected from hydrogen and C1-C4 alkyl, and R8atogether with the nitrogen to which it is bound and R7forms a fused 6-membered ring having one nitrogen heteroatom, two nitrogen heteroatoms, or one nitrogen heteroatom and one oxygen heteroatom, wherein each nitrogen heteroatom is optionally substituted by 1, 2, 3, or 4 C1-C4 alkyl substituents;or R8a, the nitrogen to which it is bound, and R7form a fused 6-membered, nitrogencontaining ring and R8b, the nitrogen to which it is bound, and R1together form a fused 6-membered, nitrogen-containing ring;with the proviso that the compounds of Formula I do not include N3, N3, N7, N7- tetraethyl-10H-phenoxazine-3,7-diamine; N3, N7-diethyl-10H-phenoxazine-3,7- diamine; N3, N3, N7, N7-tetramethyl-10H-phenoxazine-3,7-diamine; and N7, N7- diethyl-N3, N3,2-trimethyl-10H-phenoxazine-3,7-diamine.

96. The method of any one of claims 1-76 or 95, wherein the nerve homing agent homing agent is selected from:

97. The method of any one of claims 1-76. wherein the nerve homing agent is a molecule of Formula (HI):wherein:R is a straight or branched alkyl chain of from 2 to 12 carbon atoms;R1is selected from the group of methyl, ethyl, n-propyl, isopropyl, — (CH2)ni — SO3, — (CH2)m— N+(CH3)3, — CH2— CH2— O— XI, — CH2— CH2— O— [CH2— CH2— O]n2— Xi, — CH2— CH2— CH2— O— XI, and —CH2—CH2— CH2— O— [CH2— CH2— CH2— O]n3— Xi,Xi in each instance is independently selected from Ci-Ce straight or branched alkyl.C2-C6straight or branched alkenyl, C1-C6straight or branched alkynyl, and — Si(C1-C4alkyl)3;nl is an integer independently selected in each instance from the group of 1, 2, 3, and 4;n2 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;n3 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;n4 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; andn5 is an integer independently selected in each instance from the group of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;with the proviso that the sum of n2+n4 is not greater than 10;with the proviso that the sum of n3+n5 is not greater than 10;with the proviso that the sum of n2+n5 is not greater than 10; andwith the proviso that the sum of n3+n4 is not greater than 10.

98. The method of any one of claims 1-76 or 97, wherein the nerve homing agent homing agent is selected from:

99. The method of any one of claims 1-76. wherein the nerve homing agent is a molecule of Formula (JI):wherein:R1is selected from hydrogen and C1-C3 alkyl;R2is selected from hydrogen and C1-C3 alkyl;R3is selected from the group of hydrogen, halogen, and C1-C3 alkyl;or, when R3is bound to the 9-position carbon of the 10H-benzo[c]phenoxazine core, R2and R3, together with the carbon atoms to which they are bonded, form a fivemembered or six-membered fused ring, the five-membered or six-memberedfused ring optionally containing a ring oxygen heteroatom, and the fivemembered or six-membered fused ring being substituted by 0, 1, 2, or 3 Ci- C3 alky l substituents; andR4and R5are each independently selected from hydrogen and C1-C3 alkyl.

100. The method of any one of claims 1-76 or 99, wherein the nerve homing agent homing agent is selected from:

101. The method of any one of claims 1-100, wherein the nerve homing agent homing agent is detected at a 450-800 nm excitation.

102. The method of any one of claims 1-101, wherein the nerve homing agent homing agent is detected at a 550-850 nm emission.

103. The method of any one of claims 1-102, wherein the nerve homing agent homing agent is detected at a 540-760 nm excitation.

104. The method of any one of claims 1-103, wherein the nerve homing agent homing agent is detected at a 605-785 nm emission.

105. The method of any one of claims 1-104, wherein the tumor homing agent and the nerve homing agent each detect a physiologic structure or cell between at least 10 pm and 25 pm in diameter.

106. The method of any one of claims 1-105, wherein the nerve homing agent detects a physiologic structure or cell between at least 10 pm and 25 pm in diameter.

107. The method of any one of claims 1-106, further comprising causing remission in, reducing, ameliorating, or ablating the cancer or vascular lesion, include remission, increasing quality of life, reducing the extent of resection, extending progression free survival, or number of surgeries required, reducing pain, or exhibiting other clinical or quality of life benefit to a patient.

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