Development of NTSR-targeting agents for imaging and therapeutic applications

Abstract

The present invention relates to a novel NTSR1 ligand SR-CP-05 for use in targeted imaging and therapy. The invention also relates to neurotensin receptor (NTSR1) specific ligands suitable for radiolabeling and comprising a neurotensin ligand, a linker and a chelating agent for binding a radioisotope. In some embodiments, compositions comprising NTSR1 specific ligands are used for imaging, diagnosis, and / or therapeutic guidance of NTSR1 positive cancers.

Description

Priority declaration

[0001] Pursuant to 35 U.S.C. §119(e), this application claims the benefit of U.S. Provisional Application No. 63 / 364,048, filed on May 3, 2022, the entire contents of which are incorporated herein by reference in their entirety. Statement on Electronic Submission of Sequence Listings

[0002] The sequence table in XML format generated and submitted on May 3, 2023, with the file name 5470-929WO_ST26.xml and a size of 2,932 bytes, is hereby incorporated by reference into the description of the present disclosure. Technical Field

[0003] The present invention relates to methods and compositions for imaging and therapy. In particular, the present invention relates to agents targeting neurotensin receptors (NTSRs), compositions thereof, and methods of imaging and therapy using the same. Background Art

[0004] Prostate cancer is the most commonly diagnosed non-cutaneous malignancy in American men and the second leading cause of cancer-related death. Although various treatments have been developed, there is still an unmet need for greatly improved prostate cancer management. For example, agents targeting the gastrin-releasing-peptide-receptor (GRPR) have shown promising diagnostic and therapeutic potential. However, in cases of aggressive prostate cancer, a significant negative correlation was found between GRPR expression and high prostate-specific antigen (PSA) values, larger tumor size, and higher Gleason score. This is problematic because it can lead to potential false-negative imaging results, especially for tumors that should not be ignored. In addition, current research results do not support GRPR as a target for advanced aggressive prostate cancer.

[0005] Prostate-specific membrane antigen (PSMA) expression has been shown to be a prognostic factor for prostate cancer recurrence. Radiotracers based on monoclonal antibodies and other PSMA ligands have been developed for PSMA imaging, and encouraging results have been achieved in detecting early recurrence of prostate cancer after treatment. However, a major limitation of PSMA is its role in some advanced prostate cancers. Although androgen-dependent prostate cancer cell lines (including LNCaP, MDA PCa2b, and CWR22Rv1) endogenously express PSMA, it is also common that some advanced prostate cancer cells (e.g., androgen-independent PC3 or DU145) do not express PSMA. Significantly, for PSMA-positive LNCaP cells, knockdown of PSMA expression increased their invasiveness by 5-fold, indicating that PSMA can be downregulated as tumors progress.

[0006] Although multiple factors may contribute to the development, progression and resistance to treatment of prostate cancer, there is increasing evidence that intra-prostate neuroendocrine-like cells play an important role in recurrent prostate cancer that becomes independent of androgen. It is worth mentioning that after long-term anti-androgen treatment, prostate cancer can become rich in neutral endocrine cell clusters (or completely composed of neutral endocrine cell clusters). Neurotensin (NTS), secreted by neuroendocrine-like prostate cells, has many physiological effects that are mainly mediated by its high-affinity receptor NTSR1. Obviously, NTSR1 was found to be expressed and activated in invasive prostate cancer cells, but not in normal prostate epithelial cells. In advanced prostate cancer, in the absence of androgens, NTSR1 is recruited as an alternative growth pathway.

[0007] In addition, lung cancer claims approximately 160,000 lives in the United States each year and is the leading cause of cancer-related deaths in the United States. Lung cancer can be further divided into non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). Despite recent advances in lung cancer treatment, the development of more effective therapeutic and prognostic methods for lung cancer management remains an urgent unmet need. NTS and its cognate receptor (NTSR1) are neuropeptide-receptor complexes that are frequently dysregulated in neoplastic processes.

[0008] When NTS binds to NTSR1, it hydrolyzes phosphatidylinositol, leading to Ca 2+Mobilization and protein kinase C (PKC) activation. NTSR1 activation leads to cell proliferation, survival, migration and invasion in specific cancer cell types. In early NSCLC, NTSR1 positivity has been found to be associated with lower patient survival. NTS is one of 73 genes overexpressed in the highly metastatic human lung cell line H460-M compared to control cells. NTSR1 inhibitors have also been found to affect the migration and proliferation of lung cancer cells NCI-H209 and H345, and inhibit tumor growth of NCI-H209. High concentrations of NTS are also present in semiclassical SCLC cells and secreted from semiclassical SCLC. In addition, many other cancer types are positive for NTSR1, including pancreatic cancer, colorectal cancer, and breast cancer.

[0009] Novel therapeutic, diagnostic, and prognostic approaches need to be developed for the management of NTSR1-positive cancers, which may complement existing approaches to achieve precision therapy. Summary of the invention

[0010] The present invention is based on the development of novel NTSR1 ligands (eg, SR-CP-05) for targeted imaging and therapy. The newly discovered SR-CP-05 ligand has high and sustained tumor uptake, with only minimal tumor washout after two days post-injection.

[0011] One aspect of the present invention is a neurotensin receptor (NTSR1) specific ligand suitable for radiolabeling and comprising a neurotensin ligand, a linker, and a chelator for binding a radioisotope. In some embodiments, the radioisotope is selected from 64 Cu, 67 Cu, 68 Ga, 177 Lu, 18 F. 90 Y. 225 Ac, 227 Th, 223 Ra, 213 Bi, 211 At 212 Pb, 212 Bi, 230 U. 226 Th and 149 Tb.

[0012] In some embodiments, the chelating agent is one or more of NOTA, DOTA, Cross Bridge Cyclam, Cross Bridge TE2A, DF Chelating Agent, DiAm-Sarcage, and derivatives thereof. In other embodiments, the linker is selected from NOTA, DOTA, Cross Bridge Cyclam, Cross Bridge TE2A, Sarcage, and derivatives thereof, polyamines, PEG, divinyl sulfone, dihaloalkyl, phenylene isothiocyanate, p-phenylene diisothiocyanate, 1,4-diazinane, phosphate, and combinations thereof.

[0013] In some embodiments, the NTSR1-specific ligand comprises Formula I: wherein n is 1 to 3; and R is a chelating agent.

[0014] In other embodiments, the NTSR1-specific ligand comprises Formula VII: wherein Y is a bond or is selected from PEG, divinyl sulfone, dihaloalkyl, phenylene isothiocyanate, p-phenylene diisothiocyanate, 1,4-diazinane, and phosphate; X is H or LR 2 ; L is selected from the group consisting of PEG, divinyl sulfone, dihaloalkyl, phenylene isothiocyanate, p-phenylene diisothiocyanate, 1,4-diazinane, and phosphate; and R 1 and R 2 It is a chelating agent.

[0015] Another aspect of the present invention is a composition comprising a NTSR1-specific ligand. In some embodiments, the composition is used for imaging, diagnosis and / or treatment guidance of NTSR1-positive cancers.

[0016] One aspect of the invention relates to a method of performing a PET scan on a subject, comprising administering to the subject a ligand, probe and / or composition of the invention.

[0017] Another aspect of the invention provides a method of imaging tissue comprising NTSR1 -positive cancer in a subject, comprising administering to the subject a ligand, probe and / or composition of the invention.

[0018] Another aspect of the invention provides a method of imaging prostate cancer in a subject comprising administering to the subject a ligand, probe and / or composition of the invention.

[0019] Another aspect of the invention provides a method of imaging lung cancer in a subject comprising administering to the subject a ligand, probe and / or composition of the invention.

[0020] Another aspect of the invention provides a method of identifying NTSR1 -positive cancer tissue in a subject, comprising performing a PET scan on the subject using a ligand, probe or composition of the invention, wherein the PET scan identifies the presence of NTSR1 -positive cancer tissue.

[0021] Another aspect of the invention provides a method of treating NTSR1 -positive cancer tissue in a subject, comprising administering to the subject a ligand, therapeutic agent or composition of the invention.

[0022] Another aspect of the present invention provides a method for removing NTSR1-positive cancer tissue in a subject, comprising: performing a PET scan on the subject using the ligand, probe and / or composition of the present invention, wherein the PET scan identifies the presence of NTSR1-positive cancer tissue; and surgically removing the identified NTSR1-positive cancer tissue, thereby removing the NTSR1-positive cancer tissue.

[0023] Another aspect of the present invention provides a method for determining the suitability of a subject having NTSR1-positive cancer, or a subject at risk of having or developing NTSR1-positive cancer, or suspected of having or developing NTSR1-positive cancer, for surgical removal of cancer tissue, comprising (a) performing a PET scan on the subject using the ligands, probes and / or compositions of the present invention, wherein the PET scan identifies the presence of NTSR1-positive cancer tissue; and (b) identifying the presence of NTSR1-positive cancer tissue, wherein the presence of NTSR1-positive cancer tissue indicates that the subject is suitable for surgical removal of cancer tissue.

[0024] Another aspect of the invention provides a method for treating NTSR1-positive cancer in a subject, comprising predicting the suitability of surgery to remove cancer tissue in a subject having NTSR1-positive cancer or a subject at risk of having or developing NTSR1-positive cancer or suspected of having or developing NTSR1-positive cancer by performing a PET scan on the subject using a ligand, probe or composition of the invention, wherein the PET scan identifies the presence of NTSR1-positive cancer tissue, and treating the NTSR1-positive cancer based on the results of the PET scan.

[0025] Another aspect of the invention provides a method for treating a disorder in which NTSR1-positive tissue is present in a subject, comprising determining the suitability of a subject having the disorder or a subject at risk of having or developing the disorder or suspected of having or developing the disorder for treatment thereof by performing a PET scan on the subject using a ligand, probe or composition of the invention, wherein the PET scan identifies the presence of NTSR1-positive tissue, and treating the disorder based on the results of the PET scan.

[0026] These and other aspects of the present invention are described in more detail in the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Concomitant and therapeutic diagnostic methods in nuclear medicine are shown. Molecular imaging uses PET probes to detect NTSR-positive lesions. Thereafter, radionuclide-based therapy will treat NTSR1-positive tumors.

[0028] Figure 2A It was shown that NTSR1 was overexpressed on prostate cancer samples, but not on normal prostate. Figure 2B NTSR1, PSMA and GRPR staining on prostate cancer specimens is shown.

[0029] FIG3A shows the use of 18 F-VS-Cys-NTS mut Figure 3B shows a micro-PET image of an athymic male nude mouse carrying a PC3 tumor. The arrow indicates the tumor. 18 F-VS-Cys-NTS mut Biodistribution in PC-3 tumor-bearing nude mice.

[0030] Figure 4 Representative images of radiolabeled NTSR1 probe are shown.

[0031] Figure 5 Shown is a Western blot analysis of NTSR1 expression in normal mouse organs and PC3 xenografts.

[0032] Figure 6 Shown is a Western blot analysis showing NTSR1 expression in human lung cancer cell lines.

[0033] Figure 7 Shown is a Western blot analysis showing NTSR1 expression in human prostate PC3, DU145, and C4-2B cells, but not in LNCaP cells.

[0034] Figure 8Shown in a side by side comparison 64 Cu-labeled SR-CP-05 outperformed NT peptide analogs and other SR142948A-based reagents (including 64 Cu-3BP-227).

[0035] Fig. 9 The general structure of SR-CP-05 based reagents is shown.

[0036] Fig.10 A reaction scheme for forming one example of a SR-CP-05 based reagent is shown.

[0037] Fig.11 The in vivo distribution pattern of the dual-modality probe (DOTA-K(Cy5.5)-Ahx-DGEA) is shown, demonstrating the difference between optical and PET imaging.

[0038] Fig.12 Figure 2 shows the in vitro distribution pattern of the dual-modality probe (DOTA-K(Cy5.5)-Ahx-DGEA), showing the difference between optical imaging and PET imaging (A kidney, B heart, C liver, D spleen, E lung, F tumor), which indicates the need to develop stable 64 Cu chelating agent.

[0039] Fig.13 Tumor accumulation of SR-CP-18 at 24 hr is shown (high contrast). DETAILED DESCRIPTION

[0040] The present invention is explained in more detail below. This specification is not intended to be an exhaustive list of all the different ways in which the present invention can be implemented or all the features that can be added to the present invention. For example, features associated with one embodiment may be incorporated into other embodiments, and features associated with a particular embodiment may be deleted from that embodiment. In addition, in view of this disclosure, many variations and supplements suggested herein that do not depart from the various embodiments of the present invention will be apparent to those skilled in the art. Therefore, the following description is intended to illustrate some specific embodiments of the present invention, rather than to exhaustively specify all permutations, combinations, and variations thereof.

[0041] Unless the context indicates otherwise, it is a specific intention that the various features of the invention described herein can be used in any combination. In addition, the invention also contemplates that in certain embodiments of the invention, any feature or combination of features shown herein can be excluded or omitted. For illustration, if the specification states that a compound comprises components A, B, and C, it is a specific intention that any one or combination of A, B, or C can be omitted and abandoned, either alone or in any combination.

[0042] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. The terms used in the description of the invention herein are only used to describe specific embodiments and are not intended to limit the invention.

[0043] All publications, patent applications, patents, and other references cited herein are incorporated by reference in their entirety for the teachings relevant to the sentence and / or paragraph in which the citation occurs. definition

[0044] The following terminology is used in the description herein and the appended claims.

[0045] The singular forms "a" and "an" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0046] In addition, when referring to measurable values ​​(e.g., the length of a polynucleotide or polypeptide sequence, the amount of dosage, time, temperature, etc.), the term "about" as used herein refers to variations of ±10%, ±5%, ±1%, ±0.5% and even ±0.1% of the specified value as well as the specified value. For example, "about X", where X is a measurable value, means variations of ±10%, ±5%, ±1%, ±0.5% and even ±0.1% of X. The ranges of measurable values ​​provided herein may include any other ranges and / or single values ​​therein.

[0047] Furthermore, as used herein, "and / or" refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as no combinations when interpreted in the alternative ("or").

[0048] The terms “comprise”, “comprises”, and “comprising” as used herein specify the presence of stated features, integers, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof.

[0049] As used herein, the transitional phrase "consisting essentially of" means that the scope of the claim should be interpreted to cover the specific materials or steps recited in the claim, as well as those materials or steps that do not materially affect the basic and novel characteristics of the claimed invention. Therefore, the use of the term "consisting essentially of" in the claims of the present invention is not intended to be interpreted as being equivalent to "comprising".

[0050] The terms "treat", "treating" or "treatment" refer to any type of action that imparts a modulating effect (e.g., which can be a beneficial effect) on a subject suffering from a disorder, disease or illness, including an improvement in the subject's condition (e.g., in one or more symptoms), a delay or reduction in the progression of the condition, and / or a change in clinical parameters, disease or condition, etc., as is well known in the art.

[0051] As used herein, the term "therapeutically effective amount" or "effective amount" refers to an amount of a composition, compound, or agent of the invention that confers a modulating effect (e.g., which can be a beneficial effect) on a subject suffering from a disorder, disease, or condition, including an improvement in the subject's condition (e.g., in one or more symptoms), a delay or reduction in the progression of the condition, the prevention or delay of the onset of the condition, and / or a change in a clinical parameter, disease, or condition, etc., as is well known in the art. For example, a therapeutically effective amount or effective amount can refer to an amount of a composition, compound, or agent that improves the condition in a subject by at least 5% (e.g., 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 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%).

[0052] As used herein, "therapeutically effective amount", "effective amount" or "therapeutic amount" refers to an amount sufficient to provide some improvement or benefit to a subject. Alternatively, a "therapeutically effective amount", "effective amount" or "therapeutic amount" is an amount that will provide some relief (alleviation), mitigation (mitigation), reduction or stabilization in at least one clinical symptom of a subject. Those skilled in the art will recognize that the therapeutic effect does not need to be complete or curative, as long as some benefit is provided to the subject. The effective amount may vary with age, the general condition of the subject, the severity of the condition being treated, the specific agent administered, the duration of treatment, the nature of any concurrent treatment (concurrent treatment), the pharmaceutically acceptable carrier used, and similar factors within the knowledge and expertise of those skilled in the art. Where appropriate, the effective amount or therapeutic amount in any individual case can be determined by a person of ordinary skill in the art by reference to relevant texts and literature and / or using routine experiments. (For example, see Remington, The Science and Practice of Pharmacy (20th ed. 2000)).

[0053] As used herein, "pharmaceutically acceptable" refers to materials that are not biologically or otherwise undesirable, i.e., materials that can be administered to an individual with the compositions of the present invention without causing substantial adverse biological effects or interacting in a deleterious manner with any other component of the composition in which it is contained. As is well known to those skilled in the art, materials will naturally be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects to the subject (see, e.g., Remington's Pharmaceutical Science; 21 st ed.2005). Exemplary pharmaceutically acceptable carriers for the compositions of the present invention include, but are not limited to, sterile pyrogen-free water and sterile pyrogen-free saline.

[0054] The terms "administering" or "administration" of a composition of the invention to a subject include any route of introducing or delivering the compound to a subject to perform its intended function (eg, use in PET imaging, eg, for surgical guidance).

[0055] "Subject" of the present invention may include any animal in need. In certain embodiments, the subject may be, for example, a mammal, a reptile, a bird, an amphibian or a fish. Mammalian subjects may include, but are not limited to, laboratory animals (e.g., rats, mice, guinea pigs, rabbits, primates, etc.), farm or commercial animals (e.g., cattle, pigs, horses, goats, donkeys, sheep, etc.), or livestock (e.g., cats, dogs, ferrets, gerbils, hamsters, etc.). In certain embodiments, mammalian subjects may be primates, or non-human primates (e.g., chimpanzees, baboons, macaques (e.g., rhesus monkeys, cynomolgus macaques, short-tailed monkeys, pig-tailed monkeys), monkeys (e.g., squirrel monkeys, night monkeys (owl monkeys), etc.), marmosets, gorillas, etc.). In certain embodiments, mammalian subjects may be humans.

[0056] A "subject in need thereof" of the methods of the present invention may be any subject known or suspected of having cancer and / or a condition for which imaging and / or surgery may provide a beneficial health effect, or a subject at increased risk of developing the same.

[0057] The "sample", "biological sample" and / or "ex vivo sample" of the present invention can be any biological material known in the art, such as biological fluids, extracts from cells, extracellular matrix separated from cells, cells (in solution or bound to a solid support), tissues, tissue homogenates, biopsy, etc.

[0058] As used herein, by "isolating" or "purifying" (or grammatical equivalents) a fragment, it is meant that the fragment is at least partially separated from at least some other components in the starting material. NTSR1-specific ligands

[0059] Based on the important functions of NTSR1, the corresponding PET agents may serve as promising prognostic markers for identifying patients with poor prognosis in early-stage disease. In addition, the role of NTS in experimental tumor growth may represent the basis for the development of specific targeted drugs for use with currently available treatments, including therapeutic radionuclide-based agents.

[0060] As used herein, the term "neurotensin receptor (NTSR1) specific ligand" and / or "NTSR1-targeting ligand" refers to a core scaffold structure that includes a NTSR1-targeting ligand, a bifunctionalized linker, and a radionuclide component (see, e.g., Fig. 9 ).

[0061] The term "neurotensin ligand" refers to a molecule that specifically binds to NTSR1.

[0062] As used herein, the term "SR-CP-05" refers to a core scaffold structure that includes a region that binds to NTSR1. In some embodiments, the NTSR1-specific ligand of the present invention may include an NTS reagent SR-CP-05, a linker, and a chelator for binding a radioisotope. In some embodiments, the bond connection between the linker (e.g., a polyamine) and the chelator is an amide or other bond. In some embodiments, the region that binds to NTSR1 is an NTS reagent. In one embodiment, the region that binds to NTSR1 is an NTS reagent SR-CP-05 and refers to a structure having Formula X:

[0063] In some embodiments, the NTSR1-targeting ligand of the present invention may include a structure having Formula I or a pharmaceutically acceptable salt thereof: wherein n is 1 to 3, and R is a chelating agent.

[0064] Non-limiting examples of chelators are NOTA, DOTA, Cross-bridge Cyclam, Cross-bridge TE2A, Sarcage and derivatives thereof (the bond connection between the linker (eg, polyamine) and the chelator may be an amide or other bond).

[0065] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-2PA-CB and comprises a structure having Formula Ia or a pharmaceutically acceptable salt thereof:

[0066] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-1PA-CB and comprises a structure having Formula Ib or a pharmaceutically acceptable salt thereof:

[0067] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-3PA-DOTA and comprises a structure having Formula Ic or a pharmaceutically acceptable salt thereof:

[0068] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-3PA-CB and comprises a structure having Formula Id or a pharmaceutically acceptable salt thereof:

[0069] In some embodiments, the NTSR1-targeting ligand of the present invention may include a structure having Formula II or a pharmaceutically acceptable salt thereof: wherein n is 1 to 3, and R is a chelating agent.

[0070] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-1PA-NOTA and comprises a structure having Formula IIa or a pharmaceutically acceptable salt thereof:

[0071] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-2PA-NOTA and comprises a structure having Formula IIb or a pharmaceutically acceptable salt thereof:

[0072] In some embodiments, the NTSR1-targeting ligand of the present invention may comprise a structure having Formula III or a pharmaceutically acceptable salt thereof: wherein n is 1 to 3, and R is a chelating agent.

[0073] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-2PA-VS-DOTA and comprises a structure having Formula IIIa or a pharmaceutically acceptable salt thereof:

[0074] In some embodiments, the NTSR1-targeting ligand of the present invention may comprise a structure having Formula IV or a pharmaceutically acceptable salt thereof: wherein n is 1 to 3.

[0075] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-2PA-Df and comprises a structure having Formula IVa or a pharmaceutically acceptable salt thereof:

[0076] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-1PA-Df and comprises a structure having Formula IVb or a pharmaceutically acceptable salt thereof:

[0077] In some embodiments, the NTSR1-targeting ligand of the present invention may comprise a structure having Formula V or a pharmaceutically acceptable salt thereof: wherein n is 1 to 3, and R is a chelating agent.

[0078] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-1PA-sarcage and comprises a structure having Formula Va or a pharmaceutically acceptable salt thereof:

[0079] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-1PA-sarcage and comprises a structure having Formula Vb or a pharmaceutically acceptable salt thereof:

[0080] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-2PA-sarcage and comprises a structure having Formula Vc or a pharmaceutically acceptable salt thereof:

[0081] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-2PA-sarcage and comprises a structure having Formula Vd or a pharmaceutically acceptable salt thereof:

[0082] In some embodiments, the NTSR1-targeting ligand of the present invention may comprise a structure having Formula VI or a pharmaceutically acceptable salt thereof: wherein n is 1 to 3, and R is a chelating agent.

[0083] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-2PA-VS-Sarcage and comprises a structure having Formula VIa or a pharmaceutically acceptable salt thereof:

[0084] In some embodiments, the NTSR1-targeting ligand of the present invention may comprise a structure having Formula VII or a pharmaceutically acceptable salt thereof: wherein Y is a bond or is selected from PEG, divinyl sulfone, dihaloalkyl, phenylene isothiocyanate, p-phenylene diisothiocyanate, 1,4-diazinane and phosphate; X is H or LR 2 L is selected from PEG, divinyl sulfone, dihaloalkyl, phenylene isothiocyanate, p-phenylene diisothiocyanate, 1,4-diazine and phosphate; and R 1 and R 2 It is a chelating agent.

[0085] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-sarcage and comprises a structure having Formula VIIa or a pharmaceutically acceptable salt thereof:

[0086] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-sarcage and comprises a structure having Formula VIIb or a pharmaceutically acceptable salt thereof:

[0087] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-CB and comprises a structure having Formula VIIc or a pharmaceutically acceptable salt thereof:

[0088] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-CB-CA and comprises a structure having Formula VIId or a pharmaceutically acceptable salt thereof:

[0089] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-CB-CB and comprises a structure having Formula VIIe or a pharmaceutically acceptable salt thereof:

[0090] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-CB-NOTA and comprises a structure having Formula VIIf or a pharmaceutically acceptable salt thereof:

[0091] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-CB-DOTA and comprises a structure having Formula VIIg or a pharmaceutically acceptable salt thereof:

[0092] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-CB-sarcage and comprises a structure having Formula VIIh or a pharmaceutically acceptable salt thereof:

[0093] In one embodiment, the NTSR1-targeting ligand of the present invention is NT-CB-sarcage and comprises a structure having Formula VIIi or a pharmaceutically acceptable salt thereof:

[0094] In some embodiments, the NTSR1-specific ligand may comprise a radioactive isotope. The radioactive isotope may be a radioactive isotope suitable for imaging and / or for therapeutic use. In other embodiments, the radioactive isotope is selected from 64 Cu, 67 Cu, 68 Ga, 177 Lu, 18 F. 90 Y. 225 Ac, 227 Th, 223 Ra, 213 Bi, 211 At 212 Pb, 212 Bi, 230 U. 226 Th and 149 Tb. In some embodiments, the NTSR1-specific ligand has a radio purity of at least 95% (eg, 95%, 96%, 97%, 98%, 99% or more).

[0095] In some embodiments, the NTSR1-specific ligand may comprise one or more chelators selected from NOTA, DOTA, cross-bridge Cyclam, cross-bridge TE2A, DF chelators, DiAm-Sarcage and derivatives thereof. In other embodiments, the NTSR1-specific ligand may comprise one or more linkers selected from NOTA, DOTA, cross-bridge Cyclam, cross-bridge TE2A, Sarcage and derivatives thereof, polyamines (e.g., urea), PEG, divinyl sulfone, dihaloalkyl, phenylene isothiocyanate, p-phenylene diisothiocyanate, 1,4-diazinane, phosphate and combinations thereof. In some embodiments, the NTSR1-specific ligand may comprise the NTS ligand SR-CP-05.

[0096] One aspect of the present invention relates to a positron emission tomography (PET) probe comprising a NTSR1-specific ligand. In one embodiment, the PET probe has a radioactive isotope, optionally wherein the radioactive isotope is 64 Cu or 68 Ga.

[0097] Another aspect of the present invention relates to a therapeutic agent comprising a NTSR1-specific ligand. In one embodiment, the therapeutic agent comprises a radioactive isotope, optionally wherein the radioactive isotope is 67 Cu, 177 Lu, 18 F. 90 Y. 225 Ac, 227 Th, 223 Ra, 213 Bi, 211 At 212 Pb, 212 Bi, 230 U. 226 Th and 149 Tb. 18 F-labeled ligand targeting NTSR1

[0098] In some embodiments, the ligand targeting NTSR1 can be 18 F labeled. From the perspective of imaging, 18 F may be the preferred PET isotope. 68 Ga-labeled PET reagents have made progress, but recently 68 The shortage of Ga isotopes and 68 Ga / 68 The substantial increase in the cost of Ge generators justifies the need to investigate alternative reagents. 68 Compared with Ga labeling reagents, 18F-labeled analogs may have several advantages, including the ability to obtain multiple Curie s of energy from existing medical cyclotrons. 18 F; ability to generate multiple doses from a single run; extended half-life to facilitate commercialization, improved ability to image late time points; and potential improved contrast / resolution due to lower positron energy and higher positron percentage. 18 F-labeled PET agents can include chelators, which would allow integration of PET imaging with radiometal-based therapies.

[0099] 18 F radiolabeling can be carried out by conventional labeling methods, including amide bond formation, oxime formation (4- 18 F-Fluorobenzaldehyde( 18 F-FBA) with hydroxylamine-derivatized NT peptide), Michael addition reaction ( 18 F-labeled maleimide synthon with thiolated NT peptide) or click chemistry ( 18 However, most of these traditional methods usually require several time-consuming radiosynthesis steps, resulting in low overall labeling yields.

[0100] Vinyl sulfone-based labeling methods can be used to create 18 Unique sulfhydryl reactivity of F-labeled NT reagents based on vinyl sulfone 18 F marking method. This method can also be used 18 F-DEG-VS (shown below) was used to detect the thiolated NTSR1 binding ligand. 18 F mark.

[0101] The labeling method based on tetrazine trans-cyclooctene can also be used 18 F labels an effective NTSR1 binding ligand. In order to obtain a reasonable labeling yield of the final product, an excess of ligand (in the range of 200 μg to 2 mg) is generally required using the above labeling method. However, some embodiments may have a lower synthetic yield at the beginning. The amount of ligand obtained is 18 F labeling may be too little (e.g., only 50 to 100 μg of ligand is obtained). In some embodiments, before optimizing the synthesis conditions, the 18 Imaging properties of F-labeled probes. Rapid and ultra-efficient methods can be used to generate 18F-labeled probes greatly speed up the tracer development process. Ultra-efficient tetrazine-based ligation (i.e., TTCO ligation) can be used. 18 F labeling method. The tetrazine ligation reaction is a bioorthogonal reaction. The reaction is based on the reverse electron demand Diels-Alder reaction between trans-cyclooctene and diaryl tetrazine, which produces N2 as the only byproduct in the subsequent retro-[4+2] cycloaddition reaction.

[0102] 18 F-labeled trans-cyclooctene can be obtained in high radiochemical yield (71%) at hundreds of mCi. The reaction between TCO and tetrazine-modified ligands can proceed extremely fast, making it a promising candidate for preparation in seconds at low micromolar concentrations. 18 An efficient method for labeling probes with F. Using peptide-based ligands, various PET reagents targeting NTSR1 can be used based on the TTCO ligation reaction. NTSR1-binding ligands can also be functionalized with tetrazines via other reactions (e.g., thiolated ligands can be reacted with tetrazine-Mal to obtain tetrazine-NT). Using this ultra-efficient labeling method, more than 10 18 F labeled probe.

[0103] The ability to combine imaging and therapeutic agents into one molecule can minimize the differences between imaging and therapeutic agents. 18 F-labeled agents targeting NTSR1 can be used for radionuclide-based therapeutic applications.

[0104] In short, based on the TTCO ligation reaction method and the SR-CP-05-based ligand, 18 F-labeling was used to construct novel agents targeting NTSR1. Metal chelators can also be included in the design of therapeutic nuclide chelation. From a commercial perspective, this design can be utilized 18 The tetrazine of SR-CP-05 derivative can be combined with 18 F-TCO reaction to obtain PET reagents. Other clickable methods can also be used. Metal-containing and metal-free (chelated to DOTA) can be prepared 18 F-labeled reagent, and the 18 The F labeling reagent can have an isolated yield of >5%.

[0105] NTSR1 binding affinity can be assessed using H1299 cells. The selected reagents can have an IC 50<20 nM. Nonspecific binding can be less than 30%. Serum and metabolic stability of PET probes can be assessed by incubation in rat serum. Serum stability of selected reagents is >80% at 1 hr after incubation. 18 The tumor targeting efficacy and in vivo kinetics of the F-labeled NTSR1 probe can be tested in a subcutaneous H1299 xenograft model. Time points may be 0.5, 1, 2, and 4 hr after injection. For each small animal PET scan, 3D regions of interest (ROIs) of tumors, heart, liver, kidneys, and muscles can be drawn on decay-corrected whole-body coronal images to obtain imaging ROI-derived percentage ID per gram of tissue (% ID / g). At the end of each scan, the animals can be sacrificed and the accuracy of non-invasive PET quantification can be assessed by direct tissue sampling. The target specificity of the tracer in vivo can be confirmed by using a blocking experiment. Tumor uptake of the selected reagent can be>5% ID / g. Nonspecific binding in vivo can be less than 40%.

[0106] PET probes can be selected based on the following criteria: labeling yield >5% (>15 mCi product can be obtained for clinical translation), good tumor uptake (>5% ID / g at 1 hr after injection), optimal tumor / background contrast (tumor / liver>1, tumor / kidney>1, tumor / muscle>10 at 1 hr after injection), good target specificity confirmed by blocking experiments (can be reduced by >60% in the blocking group). SR-CP-05-based PET probes can be used in conjunction with NTSR1-based 68 Ga or 64 Side-by-side comparison of reagents for Cu.

[0107] Bifunctionalized linkers can be used to modify the pharmacokinetics of radiotracers and reduce uptake in the liver. For example, PEGylation can be performed to increase the water solubility of imaging probes to reduce their liver uptake. Short PEG and long PEG (PEG n , n is 2 to 10) can be used for imaging probe construction. Polysaccharide-based linkers can also be used to improve water solubility. After the bifunctionalized linkers are introduced into the peptides, their NTSR-1 binding affinity can be determined.

[0108] PET probes and / or therapeutic agents can be combined with 68 Ga or 64Cu's reagents were compared side by side to fully characterize their ability to quantify NTSR1 expression in vivo. Various lung cancer models, including H1299, H1975, H23, H226, and H460, can be imaged. All of these are NSCR1-positive NSCLC cells. Of note, H1299, H1975, and H23 originate from adenocarcinoma NSCLCs, while H226 and H460 originate from squamous cell carcinoma and large cell carcinoma, respectively. For comparison, H358 and H1973 (NTSR1-negative NSCLCs) can be used as negative controls. Western immunoblotting and FACS analysis can be performed on the above lung cancer tumors to determine NTSR1 expression levels. The corresponding xenograft models can be imaged using PET probes and / or therapeutic agents. ROIs can be drawn to obtain tumor uptake, expressed as percent ID per gram of tissue (%ID / g). Western immunoblotting analysis can be performed on tumor tissues to characterize the relative expression level of NTSR1, which can then be correlated with PET-derived tumor uptake using GraphPad software. To qualitatively assess the expression of NTSR1 in each tumor tissue, immunofluorescence staining can be performed on H1299, H1975, H23, H226, and H460 xenograft models. DAPI and CD31 staining can also be performed. Through these studies, the best imaging agent for NTSR1 can be selected for further evaluation in non-human primates. In some embodiments, the selection criteria can be: tumor / liver>1, tumor / kidney>1, tumor / muscle>10 at 1 hr after injection in the NTSR1-positive tumor model. Tumor uptake may be associated with NTSR1 expression in different tumor types. Tumor uptake values ​​can be considered together with contrast. If the contrast shows no significant difference, the one with higher tumor uptake can be selected.

[0109] PET probes and / or therapeutic agents can be evaluated in non-human primates (e.g., rhesus monkeys). Dynamic scanning can be performed over a 2-hour period following intravenous (iv) injection of a radiolabeled agent. 68 Ga, static scanning can also be performed at 3 hr; if the reagent is marked with 18 F, a static scan can be performed at 4 hr. 64Cu, static scans can be performed at 4 and 24 hr. PET probes and / or therapeutic agents may allow (1) characterization of the pharmacokinetics, biodistribution, and metabolic stability of radiolabeled PET agents following brief intravenous (iv) infusions; (2) estimation of dosimetry data; and (3) specification of clinical PET / CT scanning protocols, including injection doses and scanning time points. In detail, rhesus monkeys (NHPs) can be maintained under 1.4 to 4% isoflurane inhalation anesthesia and artificial ventilation. Two intravenous catheters can be used, one for tracer administration and the other for sampling of blood radioactivity concentrations. CT transmission scans can be obtained. Then, a radiolabeled PET agent targeting NTSR1 can be provided intravenously (3-5 mCi), and a 120-min dynamic PET scan can be performed. Scans can be performed with and without fasting to compare uptake and contrast differences. Serial venous blood samples (0.2 to 0.5 ml) can be drawn before injection and at 0.5, 5, 30, 60, and 90 min after injection to determine metabolic stability and blood uptake. Body temperature, heart rate, ECG, pCO2, pO2, SaO2, and blood pressure can be monitored throughout the study. Urine specimens for HPLC metabolite analysis can be collected at the end of the whole-body scan. PET scan data can be analyzed, including volumetric region of interest (ROI) analysis and tissue extract time-activity curves (TAC) and steady-state standardized uptake values ​​(SUV); quantitative analysis of plasma TAC and HPLC data to determine the relationship between TAC of circulating PET agent and its metabolites and time; and calculated cumulative activity of normal organs / tissues.

[0110] Plasma and urine samples can be tested for radiolabeled agents and labeled metabolites. Blood samples can be collected and immediately centrifuged at 14,000 rpm for 5 minutes. Then, 100 μL of 50% TFA in PBS is added to the upper serum solution and then centrifuged for 5 minutes. The upper solution can be injected for HPLC analysis. Urine can be filtered and then used for HPLC analysis.

[0111] The distribution of absorbed radiation dose can be calculated according to the MIRD method, which assumes that the integrated activity of each source organ is known. Source organs observed in which PET agents may be concentrated include the bladder, kidneys, and liver. Other organs whose anatomical boundaries can be identified using a combination of PET scans, attenuation scans, and control CT scans can be used to complete other source organs (brain, lower large intestine, stomach, blood, heart wall, lungs, pancreas, red bone marrow, spleen). Organs for which PET uptake above background is not observed and for which boundaries cannot be demarcated can be considered background and the remaining level of cumulative activity assigned.

[0112] Nonhuman primates may be closely monitored for the development of toxicity. Metabolic studies may be performed, including blood chemistry panels (electrolytes, glucose, calcium, phosphorus, magnesium, bilirubin, albumin, total protein, AST, ALT, ALP) to assess potential changes in liver and kidney function.

[0113] By using 10 animals per group for reagent comparison, statistically significant data can be obtained. For unpaired data between two groups, statistical differences between imaging groups at different time points and from different methods can be evaluated by ANOVA and Student's t test. The significance level can be set to P<0.05. If P>0.05, Tukey-Kramer multiple comparison post hoc test can be performed. Power calculation is based on the following assumptions: the significant reduction of imaging signal (or tumor size) is 20%, which can be designed to be detected using a one-tailed (two-group) t test with α set to 0.05. The standard deviation within the group can be set to 15% of the mean value. If there are 10 animals in each group, the power can be 0.89. The smaller group reduction of imaging signal (or tumor size) is 10%, and 10 animals in each group can have enough power (0.70). How to use

[0114] Other aspects of the invention relate to methods of imaging, diagnosing and / or guiding treatment of NTSR1-positive cancers or conditions of NTSR1-positive tissues using the compounds of the invention. In some embodiments, the NTSR1-positive cancer is a cancer from a tissue selected from the group consisting of the prostate, liver, kidney, spleen, bladder, parotid gland, lacrimal gland, submandibular gland, small intestine, nasal mucosa, esophageal mucosa, vocal cords, gallbladder, bile duct, trachea, lung, breast, mediastinal lymph nodes, axillary lymph nodes, inguinal lymph nodes, gynecomastia, sympathetic ganglia, and any combination thereof.

[0115] One aspect of the invention relates to a method of performing a PET scan on a subject, comprising administering to the subject a ligand, probe and / or composition of the invention.

[0116] Another aspect of the invention provides a method of imaging tissue comprising NTSR1 -positive cancer in a subject, comprising administering to the subject a ligand, probe and / or composition of the invention.

[0117] Another aspect of the invention provides a method of imaging prostate cancer in a subject comprising administering to the subject a ligand, probe and / or composition of the invention.

[0118] Another aspect of the invention provides a method of imaging lung cancer in a subject comprising administering to the subject a ligand, probe and / or composition of the invention.

[0119] Another aspect of the invention provides a method of identifying NTSR1 -positive cancer tissue in a subject, comprising performing a PET scan on the subject using a ligand, probe or composition of the invention, wherein the PET scan identifies the presence of NTSR1 -positive cancer tissue.

[0120] Another aspect of the invention provides a method of treating NTSR1 -positive cancer tissue in a subject, comprising administering to the subject a ligand, therapeutic agent or composition of the invention.

[0121] Another aspect of the present invention provides a method for removing NTSR1-positive cancer tissue in a subject, comprising: performing a PET scan on the subject using the ligand, probe and / or composition of the present invention, wherein the PET scan identifies the presence of NTSR1-positive cancer tissue; and surgically removing the identified NTSR1-positive cancer tissue, thereby removing the NTSR1-positive cancer tissue.

[0122] In some embodiments, the subject has, is at risk of, or is suspected of having or developing a cancer from a tissue selected from the group consisting of: prostate, liver, kidney, spleen, bladder, parotid gland, lacrimal gland, submandibular gland, small intestine, nasal mucosa, esophageal mucosa, vocal cords, gallbladder, bile duct, trachea, lung, breast, mediastinal lymph nodes, axillary lymph nodes, inguinal lymph nodes, gynecomastia, sympathetic ganglia, and any combination thereof.

[0123] Another aspect of the present invention provides a method for determining the suitability of a subject having NTSR1-positive cancer, or a subject at risk of having or developing NTSR1-positive cancer, or suspected of having or developing NTSR1-positive cancer, for surgical removal of cancer tissue, comprising (a) performing a PET scan on the subject using the ligands, probes and / or compositions of the present invention, wherein the PET scan identifies the presence of NTSR1-positive cancer tissue; and (b) identifying the presence of NTSR1-positive cancer tissue, wherein the presence of NTSR1-positive cancer tissue indicates that the subject is suitable for surgical removal of cancer tissue.

[0124] Another aspect of the invention provides a method for treating NTSR1-positive cancer in a subject, comprising predicting the suitability of surgery to remove cancer tissue in a subject having NTSR1-positive cancer or a subject at risk of having or developing NTSR1-positive cancer or suspected of having or developing NTSR1-positive cancer by performing a PET scan on the subject using a ligand, probe or composition of the invention, wherein the PET scan identifies the presence of NTSR1-positive cancer tissue, and treating the NTSR1-positive cancer based on the results of the PET scan.

[0125] In some embodiments, treating NTSR1-positive cancers comprises administering a ligand, therapeutic agent or composition of the invention. The treatment may be in addition to or in place of standard cancer treatments, including chemotherapy, immunotherapy, radiation therapy and surgery.

[0126] Another aspect of the invention provides a method for treating a disorder in which NTSR1-positive tissue is present in a subject, comprising determining the suitability of a subject having the disorder, or a subject at risk of having or developing the disorder, or suspected of having or developing the disorder, for treatment thereof by performing a PET scan on the subject using a ligand, probe or composition of the invention, wherein the PET scan identifies the presence of NTSR1-positive tissue, and the disorder is treated based on the results of the PET scan.

[0127] In some embodiments, treating a disorder comprises administering a ligand, therapeutic agent, or composition of the invention. In some embodiments, the disorder is a cancer from a tissue selected from the group consisting of the prostate, liver, kidney, spleen, bladder, parotid gland, lacrimal gland, submandibular gland, small intestine, nasal mucosa, esophageal mucosa, vocal cords, gallbladder, bile duct, trachea, lung, breast, mediastinal lymph nodes, axillary lymph nodes, inguinal lymph nodes, gynecomastia, sympathetic ganglia, and any combination thereof.

[0128] In some embodiments, the subject is a preoperative subject. In other embodiments, the subject is an intraoperative subject (e.g., wherein the subject is undergoing surgery).

[0129] In some embodiments, treating a disorder of the invention may include surgically removing at least a portion of the identified NTSR1-positive tissue and / or administering an anti-cancer therapeutic, such as, for example, a ligand, therapeutic agent, or composition targeting a ligand of NTSR1, a chemotherapeutic agent, an immunotherapeutic agent, or any combination thereof.

[0130] In some embodiments of the methods of the invention, performing a PET scan on a subject using a ligand, probe and / or composition of the invention can include administering about 1 to about 15 mCi of the ligand, probe and / or composition.

[0131] In some embodiments, administration is by intravenous injection. In some embodiments, the ligand, PET probe and / or composition has a tumor to muscle ratio (T / M) of at least 3 or more at about 1 hour after injection, optionally, at least 10 or more tumor to muscle ratio (T / M) at about 1 hour after injection. In some embodiments, the ligand, probe, therapeutic agent and / or composition makes a major organ transparent after 3 hours or less. In some embodiments, the major organ includes one or more organs selected from the liver, kidney, muscle and any combination thereof. Subjects, drug formulations and administration methods

[0132] The ligands targeting NTSR1 according to the present invention are used for veterinary and medical applications. Suitable subjects include birds and mammals. The term "birds" as used herein includes, but is not limited to, chickens, ducks, geese, quail, turkeys, pheasants, parrots, parakeets, etc. The term "mammals" as used herein includes, but is not limited to, humans, non-human primates, cattle, sheep, goats, horses, cats, canines, rodents, lagomorphs, etc. Human subjects include neonates, infants, adolescents, and adults.

[0133] In a specific embodiment, the present invention provides a pharmaceutical composition comprising a ligand targeting NTSR1 of the present invention in a pharmaceutically acceptable carrier, optionally, other medicinal agents, pharmaceutical agents, stabilizers, buffers, carriers, adjuvants, diluents, etc. For injection, the carrier is usually a liquid. For other methods of administration, the carrier can be solid or liquid. For inhalation administration, the carrier will be inhalable and, optionally, can be in the form of solid or liquid particles.

[0134] "Pharmaceutically acceptable" refers to a material that is non-toxic or otherwise undesirable, ie, the material can be administered to a subject without causing any adverse biological effect.

[0135] Another aspect of the invention is a method of administering a ligand targeting NTSR1 to a subject. The ligand according to the invention may be administered to a human subject or animal in need thereof by any means known in the art. Optionally, the ligand targeting NTSR1 is delivered in a therapeutically effective dose in a pharmaceutically acceptable carrier.

[0136] The dosage of a ligand targeting NTSR1 administered to a subject depends on the mode of administration, the disease or condition to be detected, treated and / or prevented, the condition of the individual subject, the particular ligand targeting NTSR1, etc., and can be determined in a routine manner.

[0137] In specific embodiments, more than one administration (eg, two, three, four, or more administrations) may be used to achieve desired dosing levels at various intervals (eg, hourly, daily, weekly, monthly, yearly, etc.).

[0138] In specific embodiments, administration can be local or systemic. For example, delivery of NTSR1-targeted ligands encompasses situations in which the NTSR1-targeted ligand is delivered to the target tissue and the NTSR1-targeted ligand is substantially retained within the target tissue (also referred to as "local distribution" or "local delivery"), as well as situations in which the NTSR1-targeted ligand is delivered to the target tissue and the NTSR1-targeted ligand is secreted into the patient's circulatory system (e.g., serum) and distributed systemically and absorbed by other tissues (also referred to as "systemic distribution" or "systemic delivery"). Delivery can also be to airway epithelial cells or any tissue affected by cancer, such as epithelial cells from the lungs, nose, ears, eyes, nervous system, and gastrointestinal and reproductive tract tissues.

[0139] Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. Alternatively, the NTSR1-targeting ligands of the invention can be administered topically.

[0140] Non-limiting examples of formulations of the invention include those suitable for oral, rectal, buccal (e.g., sublingual), vaginal, parenteral (e.g., subcutaneous, intramuscular (including skeletal muscle, cardiac muscle, diaphragm, and smooth muscle), intradermal, intravenous, intraperitoneal), topical (i.e., skin and mucosal surfaces, including airway surfaces), intranasal, transdermal, intraarticular, intracranial, intrathecal, cerebrospinal and inhalation administration, otic administration, ocular administration, administration to the liver via intraportal delivery, and direct organ injection (e.g., into the liver, into a limb, into the brain or spinal cord for delivery to the central nervous system, into the pancreas, or into a tumor or tissue surrounding a tumor). In any particular case, the most appropriate route will depend on the nature and severity of the condition to be detected or treated, as well as the nature of the particular compound being used.

[0141] For injection, the carrier is typically a liquid, such as sterile pyrogen-free water, pyrogen-free phosphate-buffered saline solution, bacteriostatic water, or Cremophor EL[R] (BASF, Parsippany, NJ). For other methods of administration, the carrier may be a solid or a liquid.

[0142] Ligands targeting NTSR1 can also be formulated for nasal, otic, or ocular administration, or otherwise administered to the lungs of a subject by any appropriate means (e.g., by inhalation of an aerosol suspension of inhalable particles comprising the compound by the subject). The inhalable particles can be liquid or solid. The term "aerosol" includes any airborne suspension that can be inhaled into the bronchioles or nasal passages. Specifically, aerosols include airborne droplet suspensions that can be prepared in a metered dose inhaler or a nebulizer or mist sprayer. Aerosols also include dry powder compositions suspended in air or other carrier gas, which can be delivered, for example, by insufflation of an inhaler device. See Ganderton & Jones, Drug Delivery to the Respiratory Tract, Ellis Horwood (1987); Gonda (1990) Critical Reviews in Therapeutic Drug Carrier Systems 6: 273-313; and Raeburn et al., J. Pharmacol. Toxicol. Meth. 27: 143 (1992). Aerosols of liquid particles containing compounds can be prepared by any suitable method known to those skilled in the art (e.g., using a pressure-driven aerosol nebulizer or an ultrasonic nebulizer). See, for example, U.S. Pat. No. 4,501,729. Similarly, aerosols of solid particles containing compounds can be prepared using any solid particle drug aerosol generator by techniques known in the pharmaceutical art.

[0143] Having described the present invention, the same will be explained in more detail in the following examples, which are for illustrative purposes only and are not intended to limit the present invention. Example 1

[0144] Issues to be resolved. The high prevalence of NTSR1 in prostate cancer makes it a promising target for concomitant imaging and therapy of prostate cancer. However, due to rapid degradation by endogenous proteases and peptidases, native NTS has a short half-life in the blood and is therefore not suitable for imaging / therapy applications. Based on stable NTS analogs, several NTSR1 PET agents have been developed that exhibit high contrast in imaging of NTSR1-positive tumor models (using 18 F and 64 Cu labeling). In the PC3 tumor model, PET reagents 18 F-VS-Cys-NTS mutThey exhibited ratios of 19.4±5.5 (tumor / muscle), 15.6±4.1 (tumor / liver), and 3.0±0.3 (tumor / kidney), respectively. However, the agent exhibited only a modest 1.3% injected dose / gram (ID / g) tumor uptake and was cleared from mice within a few hours. Peptide-based NTSR1 agents may be suitable for imaging purposes but not for therapeutic applications. NTSR1 therapeutic diagnostic agents can have both high contrast and prominent and sustained tumor uptake. SR48692 or its derivative SR142948 can selectively inhibit NTSR1. SR48692 or its derivative SR142948 is a class of non-peptide antagonists that preferentially bind to NTSR1 and inhibit its downstream signaling pathways.

[0145] Solution. 1) Discovery of a novel NTSR1 ligand, SR-CP-05, for targeted imaging and therapy. Introduction of a cross-linked polyamine into SR142948A (leading to the novel agent SR-CP-05) resulted in a 10-fold increase in tumor uptake compared to peptide-based agents. Although tumor-to-muscle contrast was slightly reduced after injection, the ratio was still >20 at 1 hr post-injection. More importantly, the agent persisted within the tumor for up to 48 hours, making it also useful for therapeutic applications.

[0146] Imaging agents targeting NTSR1 are constructed based on NTS peptide derivatives. Although high tumor to background contrast can be obtained, the absolute tumor uptake from subjects is moderate, with rapid clearance. In addition, these NTSR1 binding peptides are generally agonists of NTSR1, which can promote tumor progression at high concentrations. Due to the limited amounts, this may not be a problem for PET imaging, but may be a potential risk in treatment. Antagonists based on SR142948A were developed, with the lead agent being 3BP-227. A side-by-side comparison of the lead agent SR-CP-05 and previously reported agents was performed. As Figure 8 As shown, SR-CP-05 exhibited 15.5% injected dose / gram (ID / g) tumor uptake (more than 10-fold higher than the peptide probe and 75% higher than 3BP-227 (8.9% ID / g)) and high contrast (tumor / muscle>20, compared to 3BP-227 tumor / muscle>3 at 1 hr post-injection). Importantly, SR-CP-05 maintained tumor uptake of >15% ID / g at 48 hr post-injection. In contrast, tumor uptake of 3BP-227 decreased to 2.3% ID / g at 24 hr post-injection. The unique distribution profile of SR-CP-05 may provide imaging and radionuclide-based therapeutic agents targeting NTSR1.

[0147] 2) The radiolabeling method is innovative. Sarcophagin (Sar) cages are used to stabilize 64 Cu chelation. Sar cages were shown to significantly increase tumor-to-background contrast, likely due to their cage-like structure and positive charge. These positively charged chelators match the positive charge of the polyamines in SR-CP-05, potentially reducing the effect of the chelator on bound ligands.

[0148] These novel agents can be used for patient screening, radionuclide-based therapy, and treatment monitoring, all of which are expected to impact the care of patients with advanced prostate cancer. In summary, the SR-CP-05 ligand may be an innovative product for the management of prostate cancer patients through imaging and therapy targeting NTSR1.

[0149] Although this example focuses on prostate cancer, it should be noted that NTSR1 is also upregulated in many other solid tumors, including lung, head and neck, colorectal, and breast cancers. Therefore, these agents can be extended to other tumor types.

[0150] NTSR1, PSMA, and GRPR expression have been evaluated in prostate cancer patient tissues. It is important to determine the NTSR1 expression profile in various prostate cancer states in patient samples. NTSR1 expression was evaluated in normal prostate and localized prostate cancer. Of the 97 prostate cancer patient samples evaluated, 94 samples showed high NTSR1 expression and 3 samples showed moderate NTSR1 expression ( Figure 2B ), whereas normal prostate samples showed low to negative NTSR1 expression ( Figure 2A ). In addition to NTSR1, the same prostate patient tissues were also stained for PSMA and GRPR expression. Of the 75 PSMA stained samples, 14 samples (18.7%) showed moderate to low PSMA expression. Of the 72 GRPR stained samples, 14 samples (19.4%) showed moderate to negative GRPR expression. It is evident that for all samples with moderate to negative PSMA and GRPR expression, NTSR1 was highly expressed. Table 1 shows representative examples (NTSR1 high, PMSA low, and GRPR moderate). These preliminary staining studies clearly demonstrate that NTSR1 can complement PSMA / GRPR for more precise prostate cancer management. Table 1 Express NTSR1 PSMA GRPR high 94(96.9％) 61(81.3％) 58(80.6％) medium 3(3.1％) 11(14.7％) 5(6.9％) Low 0 3(4％) 7(9.7％) Negative 0 0 2(2.8％) total# 97 75 72

[0151] Stable NTS peptide derivatives and constructed NTSR1 PET reagents have been developed. Designed for 18 F-labeled Cys-NTS analog (Cys-NTS mut:Cys-pipGly-Pro-pipAmGly-Arg-Pro-Tyr-tBuGly-Leu-OH). Figure 3 shows a representative example. In the PC3 tumor model, 18 F-DEG-VS-NTS showed good tumor uptake (1.3 ± 0.1% ID / g) and low background. Biodistribution studies were performed at 3 hr after injection, showing tumor to muscle, liver, and kidney ratios of 19.4 ± 5.5, 15.6 ± 4.1, and 3.0 ± 0.3, respectively. The cold NTS peptide successfully blocked the PC3 tumor 18 Tumor uptake of F-DEG-VS-NTS indicated the receptor specificity of this imaging agent. Specifically, 18 F-DEG-VS-NT showed low background in major tissues / organs including blood, muscle, liver, and kidney. Despite its high contrast, its tumor retention was low.

[0152] Most normal tissues were shown to have low NTSR1 expression. The presence of NTSR1 in normal tissues may result in high background uptake and low contrast. Therefore, western blot analysis was performed to assess the relative expression of NTSR1 in tumors and normal organs. Mouse brain was used as a positive control. Figure 5 As shown in Figure 2, NTSR1 protein was very low or undetectable in most organs, including spleen, liver, lung, pancreas, muscle, leukocytes, and platelets. NTSR1 was mainly observed in the intestine, brain, and PC-3 tumors. Figure 3A-3B As shown in Figure 2, the brain has minimal uptake of tracers targeting NTSR1 because the agent does not cross the blood-brain barrier (BBB). Although the small intestine shows high NTSR1 expression, its tracer uptake is only 1 / 6 of that in PC-3 tumors based on biodistribution experiments ( Figure 3A-3B ). Therefore, there should be less concern about radiation to this sensitive organ. Overall, based on these preliminary data, background uptake should not be a major concern.

[0153] Various advanced prostate cancer cell lines have high NTSR1 expression. Induction of NTS and NTSR1 as alternative growth pathways in advanced prostate cancer. To validate that NTSR1 would be an effective target in advanced prostate cancer, Western blot analysis was performed on four prostate cancer cell lines: LNCaP (derived from lymph node metastasis), C4-2B (bone metastatic subline derived from LNCaP), DU145 (intermediate metastatic potential), and PC-3 (high metastatic potential, PSMA negative). Figure 7As shown, LNCaP cells showed minimal NTSR1 as parental cells, but showed high NTSR1 expression when it became the bone metastatic subline C4-2B. Prostate cancer cells PC3 showed prominent NTSR1 expression, but low PSMA expression. Clearly, NTSR1-targeted imaging and therapy may be an excellent supplement to current prostate cancer management.

[0154] SR-CP-05 was found to be highly suitable for therapeutic applications. Although the NT analog NTS mut and NTS 20.3 Peptides have shown promising tumor imaging results, but the absolute tumor uptake value is only about 1.5% ID / g, which is rapidly cleared at 3 to 4 hr after injection. Clearly, the rapid clearance may be acceptable from an imaging perspective, but these ligands are not suitable for therapeutic applications. After extensive searching and modification of various NTSR1 ligands, it was found that after introducing a cross-linked polyamine to SR142948A, the resulting reagent SR-CP-05 showed 15.6% ID / g tumor uptake ( Figure 8 ), which was more than 10-fold higher than the peptide probe in a side-by-side comparison study and 75% higher than 3BP-227 (SR142948A-based reagent). Importantly, high contrast was obtained at 1 hr post-injection (tumor / background>20), and washout was minimal even after 48 hr post-injection. In contrast, the peptide-based reagent showed significant washout even at 4 hr post-injection; and 3BP-227 exhibited much lower tumor-to-background ratios (only about 3 to 4) at early time points ( Figure 8 ), and in a side-by-side comparison only 1 / 3 of the tumor uptake was maintained at 24 hr post-injection. Clearly, SR-CP-05 represents a very promising ligand for imaging and therapeutic applications. In conclusion, NTSR1 was demonstrated to be an effective target for prostate cancer management and was used as a lead agent, with SR-CP-05 showing high and sustained tumor uptake.

[0155] A ligand targeting NTSR1 was constructed based on SR-CP-05. Although not wishing to be bound by theory, regarding the greatly enhanced tumor uptake and retention of SR-CP-05, it is believed that the positive charge of the cross-linked polyamine increases the interaction of SR-CP-05 with the cell membrane, increasing the chance of binding to NTSR1 and retention. The radiolabeled radioactive agent targeting NTSR1 can be divided into three parts: a ligand targeting NTSR1, a bifunctionalized linker, and a radionuclide component ( Fig. 9 ). Example 2

[0156] Issues to be resolved.NTSR1 is a target for concomitant imaging and therapy in lung cancer. From an imaging perspective, the development of PET agents to acquire NTSR1 cellular expression profiles or individual tumor “fingerprints” could be used for patient screening and treatment monitoring in patients with non-small cell lung cancer (NSCLC). PET agents targeting NTSR1 could allow identification of patients with poor prognosis within a defined stage, which could lead to more appropriate postoperative treatment (personalized therapy) accordingly. PET probes need to have low background in the abdominal region, including the liver and kidneys, in order to effectively detect small lesions surrounding this region. A number of radiolabeled NTS peptide analogs have been developed as valuable tools for imaging and therapy of NTSR1-positive tumors. Several NTSR1 PET agents have been developed, including those labeled via a tetrazine-trans-cyclooctene (TTCO) ligation reaction. 18 F reagent), which showed high contrast (moderate tumor uptake with minimal accumulation in the upper torso and most abdominal regions, including the kidneys and liver). Detailed information is shown in the results ( Figure 3A-3B , 4 ). Despite the encouraging results in these imaging studies, several challenges need to be addressed to develop these probes into clinically useful probes. First, NTS is a tridecapeptide that has a short half-life in the blood due to rapid degradation by endogenous peptidases. Changes have been introduced to help stabilize the bonds between Arg8-Arg9, Pro10-Tyr11, and Tyr11-Ile12 and provide metabolic stability. For example, the NT-20.3 peptide was demonstrated to be an NTSR1 ligand with good stability for diagnostic or therapeutic purposes. However, the resulting reagent is still cleared from mice within a few hours.

[0157] NTSR1 can be a target for lung cancer progression. Therefore, imaging and therapy targeting NTSR1 can become an important imaging / therapeutic strategy for managing lung cancer. Imaging agents targeting NTSR1 are constructed based on NTS peptide derivatives. Although high tumor to background contrast can be obtained, the absolute tumor uptake from the subject is moderate, with a rapid clearance rate. In addition, these NTSR1-binding peptides are generally agonists of NTSR1, which can promote tumor progression at high concentrations. This may not be a problem for PET imaging due to the limited administration amount, but it does increase the potential risk. Reagents based on SR142948A have been developed, with the lead agent being 3BP-227. There is a need to develop imaging and therapeutic agents targeting NTSR1 that have high and sustained tumor uptake and relatively low background.

[0158] Solution.1) Discovery of a novel NTSR1 ligand, SR-CP-05, for targeted imaging and therapy. Introduction of a cross-linked polyamine into SR142948A (leading to the novel agent SR-CP-05) resulted in a 10-fold increase in tumor uptake compared to peptide-based agents. Although tumor to muscle contrast was slightly reduced after injection, the ratio was still >20 at 1 hr post-injection. More importantly, the agent persisted within the tumor for up to 48 hours, making it also useful for therapeutic applications. A side-by-side comparison was performed between SR-CP-05 and previously reported agents. Figure 8 As shown, tumor uptake of SR-CP-05 showed 15.5% ID / g tumor uptake, which is more than 10 times higher than the peptide probe and 75% higher than 3BP-227. Importantly, SR-CP-05 also achieved high contrast (tumor / muscle>20) at 1 hr after injection, and washout was minimal after 48 hr after injection (compared to 3BP-227 tumor / muscle>3 at 1 hr after injection). In addition, both peptide-based agents and 3BP-227 showed significant washout at 24 hr after injection. The uptake of 3BP-227 also decreased from 8.9% ID / g at 1 hr after injection to 2.3% ID / g at 24 hr after injection; while the tumor uptake of SR-CP-05 at both time points remained>15% ID / g. The unique distribution curve of SR-CP-05 provides imaging and radionuclide-based therapy targeted to NTSR1.

[0159] These novel agents can be used for lung cancer prognosis, treatment monitoring, and radionuclide-based therapy, all of which may impact the care of NSCLC patients. NTSR1 has been found to be upregulated in many other solid tumors. Therefore, the agents described herein may be extended to other tumor types, including prostate cancer, head and neck cancer, colorectal cancer, and breast cancer.

[0160] A simple method for preparing NT peptide derivatives was developed. 18 Robustness of F-NTS reagent 18 F marking method. As one of the commonly used PET radioisotopes, 18 F can be easily prepared in large quantities on a medical cyclotron and has an ideal half-life of 110 min for imaging applications. 18 The half-life of F is sufficient to extend the synthesis, delivery, and imaging process over several hours with limited radiation exposure to the patient. A platform technology based on vinyl sulfone and TTCO ligation reactions was developed, both of which are 18 F PET probe construction is very efficient. 18 F labeling method, a stable NTS analogue was designed with the 18 F-labeled Cys (Cys-NTS mut:Cys-pipGly-Pro-pipAmGly-Arg-Pro-Tyr-tBuGly-Leu-OH). 18 Both F-VS and TTCO ligation reactions have been used to construct NTSR1-targeting 18 F-labeled PET reagent. Figure 3A-3B Representative examples are shown. In the NTSR1-positive PC3 tumor model, 18 F-DEG-VS-NTS showed good tumor uptake (1.3 ± 0.1% ID / g) and low background. Biodistribution studies were performed at 3 hr after injection, which showed that the ratios of tumor to muscle, liver, and kidney were 19.4 ± 5.5, 15.6 ± 4.1, and 3.0 ± 0.3, respectively. 18 The uptake of F-DEG-VS-NTS was effectively blocked, thus clearly demonstrating the receptor specificity of this imaging agent. Specifically, 18 F-DEG-VS-NT showed low background in major tissues / organs including blood, muscle, liver and kidney. The very low background in normal tissues allowed the detection of small tumor metastases (especially around the liver area) by PET imaging.

[0161] PET agents targeting NTSR1 were developed. In addition to targeting NTSR1 18 In addition to the F-VS labeled reagent, other labeling methods and radioisotopes for labeling NTS peptide analogs were explored. Various radiolabeled NTS peptides were synthesized as lead compounds and screened for their imaging properties and target specificity. Selected chemical structures are shown below, along with representative imaging targets for NTSR1. Figure 4 As shown. Overall, these peptide-based imaging agents exhibit high tumor-to-background contrast and rapid clearance from the background (including the kidney). Limitations are moderate tumor uptake and rapid clearance profiles, which would make them unsuitable for radionuclide-based therapeutic applications.

[0162] Most normal tissues have been shown to have low NTSR1 expression, eliminating potential concerns about tumor target selectivity. The presence of NTSR1 in normal tissues may result in high background uptake and low contrast. Therefore, western blot analysis was performed to assess the relative expression of NTSR1 in tumors and normal organs ( Figure 5 ). Mouse brain was used as a positive control. Figure 3A-3BAs shown in Figure 2, NTSR1 protein was very low or undetectable in most organs, including spleen, liver, lung, pancreas, muscle, leukocytes, and platelets. NTSR1 was mainly observed in the intestine, brain, and NTSR1-positive PC-3 tumors. Figure 3A-3B and 4 As shown in Figure 2, the brain has minimal tracer uptake targeting NTSR1 because the compounds of the present invention do not cross the blood-brain barrier. Although the small intestine shows high NTSR1 expression, its tracer uptake is only 1 / 6 of that in NTSR1-positive tumors according to biodistribution experiments ( Figure 3A-3B ). Therefore, there should be less concern about radiation to this sensitive organ. Overall, the peptide-based lead compound demonstrated very clean background in the upper torso and most of the abdominal region, clearly justifying its use for the detection of NTSR1-positive lung cancer and metastases in these areas. Based on these data, background uptake is not a major concern ( Figure 3A-3B and 5 ).

[0163] Various lung cancer cell lines have high NTSR1 expression. NTS and NTSR1 may play a key role in a large proportion of lung cancer patients. It is reported that H1299, H1975, H23, H226 and H460 are all NTSR1 positive. H1299 has been confirmed to be a lung cancer cell line with high NTSR expression. Figure 6 As shown, western blotting confirmed that H23 and H226 cell lines have high NTSR1 expression. Specifically, imaging and therapy targeting NTSR1 may be an excellent supplement to current lung cancer management.

[0164] SR-CP-05 was found to be well suited for imaging and therapeutic applications. Although the NT analog NTS mut The NTSR1 and NTS20.3 peptides have shown promising tumor imaging results, but the absolute tumor uptake values ​​were only about 1.5% ID / g, which was rapidly cleared at 3 to 4 hr post-injection. Clearly, the rapid clearance may be acceptable from an imaging perspective, but these ligands are not suitable for therapeutic applications. It was found that modifications to various NTSR1 ligands (e.g., introduction of cross-linked polyamines to SR142948A) resulted in the resulting reagent SR-CP-05 showing 15.6% ID / g tumor uptake ( Figure 8), which was more than 10-fold higher than the peptide probe in a side-by-side comparison study and 75% higher than 3BP-227 (a previously reported SR142948A-based agent). Importantly, high contrast was obtained at 1 hr post-injection (tumor / background>20), and washout was minimal even after 48 hr post-injection. In contrast, the peptide-based agents showed significant washout even at 4 hr post-injection; and 3BP-227 exhibited much lower tumor-to-background ratios (only about 3 to 4) at early time points ( Figure 8 ), and in a side-by-side comparison only one-third of the tumor uptake was maintained at 24 hr post-injection. Clearly, SR-CP-05 represents a very promising ligand for imaging and therapeutic applications. The relatively high contrast at early time points also makes it feasible to develop SR-CP-05-based 18 F-labeled reagents are now available for PET imaging.

[0165] Two types of agents targeting NTSR1 were developed that either exhibited high contrast (peptide-based agents, which had moderate tumor uptake but high contrast: minimal accumulation in the upper torso and most abdominal regions, including the kidneys and liver) or showed high and sustained tumor uptake (SR-CP-05-based agents).

[0166] Development of ligands targeting NTSR1 based on SR-CP-05. Although not wishing to be bound by theory, regarding the greatly enhanced tumor uptake and retention of SR-CP-05, it is believed that the positive charge of the cross-linked polyamine increases the interaction of SR-CP-05 with the cell membrane, which as a result increases the chance of binding to NTSR1 and retention. The novel radiolabeled radioactive agent targeting NTSR1 can be divided into three parts: a ligand targeting NTSR1, a bifunctionalized linker, and a radionuclide component ( Fig. 9 ).

[0167] Sar was demonstrated to be an excellent chelator for Cu labeling. Copper 64(t 1 / 2 =12.7hr) decays by β+ (20%) and β emission (37%) as well as electron capture (43%), making it very suitable for radiolabeling proteins, antibodies and peptides for PET imaging (β+) and therapy (β+ and β-). 64 Limitations of chelators used with Cu include 64The significant loss of Cu leads to high uptake in the liver. To overcome this limitation, a new class of bifunctionalized chelators (BFCs) based on the hexaazamacrobicyclic sarcophagine cage SarAr (1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexaazabicyclo[6.6.6]-eicosane-1,8-diamine) was synthesized. These ligands coordinate metal ions (e.g., Cu) within multiple macrocycles comprising the Sar cage structure. 2+ ), resulting in a very stable complex that is inert to the dissociation of metal ions. The functionalization method of Sar cages has been improved by direct alkylation (SN2) reaction. 64 Cu labeling was shown to be superior. A multimodal probe was constructed (shown below) and it was demonstrated that DOTA would provide misleading information due to its stability issues.

[0168] like Fig.11 and Fig.12 As shown, both PET and optical images demonstrate tumor uptake. However, the kidneys had much higher tracer uptake compared to the liver, while the pattern on PET imaging was the opposite (Cu-64 fell off DOTA and resulted in high liver uptake). This study clearly demonstrates that Sar may be a preferred ligand for Cu chelation in in vivo applications.

[0169] The multifunctionalized Sar cage was demonstrated to have great potential for efficiently constructing PET / optical dual-modality probes. It was also clearly shown that neither the Sar cage nor Cu significantly quenched the light signal from the fluorescent dye. This combination of PET imaging and fluorescence may allow fluorescence-guided surgery based on PET / FL constructs made of heterofunctionalized Sar cages.

[0170] The SR-CP-05 distribution profile in vivo was shown to be suitable for imaging and therapeutic applications. However, its liver uptake was still relatively high. Changing the PK linker, SR-CP-18 (NT-1PA-CB) (shown below) showed primarily tumor accumulation (high contrast) at the 24 hr time point ( Fig.13 ), but tumor uptake was less than half of that of SR-CP-05. Specifically, the distribution of SR-CP-05 derivatives can be tuned via linkers. 64 Cu / 67 Cu is a true theranostic radionuclide pair. 68 Ga(t 1 / 2 = 68min, imaging) and 177 Lu(t 1 / 2 =6.7 days, treatment) for different64 Cu(t 1 / 2 = 12.7h, imaging) and 67 Cu(t 1 / 2 =61.8h, half-life between treatment 64 Cu and 67 Cu matches better. In addition, since they are both Cu 2+ ions, thus eliminating the differences caused by using different metals.

[0171] All publications, patents, and patent applications are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0172] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent that certain changes and modifications may be practiced within the scope of the foregoing list of embodiments and the appended claims.

Claims

1. A neurotensin receptor (NTSR1) specific ligand suitable for radiolabeling, the NTSR1 specific ligand comprising: Neurotensin ligand; Connectors; and Chelating agents used to bind radioactive isotopes.

2. The NTSR1-specific ligand according to claim 1, further comprising a radioactive isotope.

3. The NTSR1-specific ligand according to claim 2, wherein the radioisotope is selected from 64 Cu, 67 Cu, 68 Ga, 177 Lu, 18 F. 90 Y. 225 Ac, 227 Th, 223 Ra, 213 Bi, 211 At 212 Pb, 212 Bi, 230 U. 226 Th and 149 Tb.

4. The NTSR1-specific ligand of claim 2 or 3, having a radioactivity purity of at least 95% (eg, 95%, 96%, 97%, 98%, 99% or more).

5. The NTSR1-specific ligand according to any one of claims 1 to 4, wherein the chelator is NOTA, DOTA, cross-bridge Cyclam, cross-bridge TE2A, DF chelator, DiAm-Sarcage or a derivative thereof.

6. The NTSR1-specific ligand according to any one of claims 1 to 5, wherein the linker is NOTA, DOTA, cross-bridge Cyclam, cross-bridge TE2A, Sarcage and its derivatives, polyamines, urea, polyethylene glycol, divinyl sulfone, dihaloalkyl, phenylene isothiocyanate, p-phenylene diisothiocyanate, 1,4-diazinane, phosphate, and combinations thereof.

7. The NTSR1 -specific ligand according to any one of claims 1 to 6, wherein the neurotensin ligand is SR-CP-05.

8. The NTSR1-specific ligand according to any one of claims 1 to 7, comprising formula I: wherein n is 1 to 3; and R is a chelating agent. 9 . The NTSR1-specific ligand according to claim 8 , wherein the chelator is NOTA, DOTA, cross-bridge Cyclam, cross-bridge TE2A, Sarcage or a derivative thereof.

10. The NTSR1-specific ligand according to any one of claims 1 to 7, comprising formula II: wherein n is 1 to 3; and R is a chelating agent.

11. The NTSR1-specific ligand according to claim 10, wherein the chelator is selected from NOTA, DOTA, cross-bridge Cyclam, cross-bridge TE2A, Sarcage, and derivatives thereof.

12. The NTSR1-specific ligand according to any one of claims 1 to 7, comprising formula III: wherein n is 1 to 3; and R is a chelating agent.

13. The NTSR1-specific ligand according to claim 12, wherein the chelating agent is selected from NOTA, DOTA, cross-bridge Cyclam, cross-bridge TE2A, DiAm-Sarcage, and derivatives thereof.

14. The NTSR1-specific ligand according to any one of claims 1 to 7, comprising formula IV: wherein n is 1 to 3.

15. The NTSR1-specific ligand according to any one of claims 1 to 7, comprising formula V: wherein n is 1 to 3; and R is a chelating agent.

16. The NTSR1-specific ligand according to claim 15, wherein the chelator is selected from NOTA, DOTA, cross-bridge Cyclam, cross-bridge TE2A, DiAm-Sarcage, and derivatives thereof.

17. The NTSR1-specific ligand according to any one of claims 1 to 7, comprising formula VI: wherein n is 1 to 3; and R is a chelating agent.

18. The NTSR1-specific ligand according to claim 17, wherein the chelator is selected from NOTA, DOTA, cross-bridge Cyclam, cross-bridge TE2A, DiAm-Sarcage, and derivatives thereof.

19. The NTSR1-specific ligand according to any one of claims 1 to 7, comprising formula VII: wherein Y is a bond or is selected from PEG, divinyl sulfone, dihaloalkyl, phenylene isothiocyanate, p-phenylene diisothiocyanate, 1,4-diazinane, and phosphate; X is H or LR 2 ; L is selected from the group consisting of PEG, divinyl sulfone, dihaloalkyl, phenylene isothiocyanate, p-phenylene diisothiocyanate, 1,4-diazinane, and phosphate; and R 1 and R 2 It is a chelating agent.

20. The NTSR1-specific ligand according to claim 19, wherein the chelating agent is independently selected from NOTA, DOTA, cross-bridge Cyclam, cross-bridge TE2A, DiAm-Sarcage, and derivatives thereof.

21. The NTSR1-specific ligand according to any one of claims 1 to 7, wherein the NTSR1-specific ligand is selected from:

22. A positron emission tomography (PET) probe comprising the NTSR1 -specific ligand according to any one of claims 1 to 21.

23. The PET probe according to claim 22, further comprising a radioactive isotope, optionally wherein the radioactive isotope is 64 Cu or 68 Ga.

24. A therapeutic agent comprising the NTSR1 -specific ligand of any one of claims 1 to 21.

25. The therapeutic agent of claim 24, further comprising a radioactive isotope, optionally wherein the radioactive isotope is 67 Cu, 177 Lu, 18 F. 90 Y. 225 Ac, 227 Th, 223 Ra, 213 Bi, 211 At 212 Pb, 212 Bi, 230 U. 226 Th or 149 Tb.

26. A composition comprising a ligand or PET probe according to any one of claims 1 to 23 and a pharmaceutically acceptable carrier.

27. A composition comprising a ligand or therapeutic agent according to any one of claims 1 to 21 or 24 to 26 and a pharmaceutically acceptable carrier.

28. Use of the ligand of any one of claims 1 to 21, the PET probe of claim 22 or 23, or the composition of claim 26 in imaging, diagnosing and / or guiding treatment of NTSR1-positive cancer.

29. Use of the ligand, probe and / or composition according to claim 28, wherein the NTSR1-positive cancer is a cancer from a tissue selected from the group consisting of: prostate, liver, kidney, spleen, bladder, parotid gland, lacrimal gland, submandibular gland, small intestine, nasal mucosa, esophageal mucosa, vocal cords, gallbladder, bile duct, trachea, lung, breast, mediastinal lymph nodes, axillary lymph nodes, inguinal lymph nodes, gynecomastia, sympathetic ganglia, and any combination thereof.

30. A method of performing a PET scan on a subject, comprising administering to the subject a ligand, PET probe or composition as claimed in any one of claims 1 to 23 or 26.

31. A method of imaging tissue comprising NTSR1 -positive cancer in a subject, comprising administering to the subject a ligand, PET probe or composition of any one of claims 1 to 23 or 26.

32. A method of imaging prostate cancer in a subject comprising administering to the subject a ligand, PET probe or composition of any one of claims 1 to 23 or 26.

33. A method of imaging lung cancer in a subject, comprising administering to the subject the ligand, PET probe or composition of any one of claims 1 to 23 or 26.

34. A method of identifying NTSR1 -positive cancer tissue in a subject, comprising performing a PET scan on the subject using the ligand, probe or composition of any one of claims 1 to 23 or 26, wherein the PET scan identifies the presence of NTSR1 -positive cancer tissue.

35. A method of treating an NTSR1 -positive cancer in a subject, comprising administering to the subject a ligand, therapeutic agent or composition of any one of claims 1 to 21, 24 to 25 or 27.

36. A method for removing NTSR1-positive cancer tissue in a subject, comprising: Performing a PET scan on a subject using the ligand, PET probe or composition of any one of claims 1 to 23 or 26, wherein the PET scan identifies the presence of NTSR1 -positive cancer tissue; and Identified NTSR1-positive cancer tissue is removed surgically to remove the NTSR1-positive cancer tissue.

37. The method of any one of claims 34 to 36, wherein the subject has, is at risk of, or is suspected of having or developing a cancer from a tissue selected from the group consisting of: prostate, liver, kidney, spleen, bladder, parotid gland, lacrimal gland, submandibular gland, small intestine, nasal mucosa, esophageal mucosa, vocal cords, gallbladder, bile duct, trachea, lung, breast, mediastinal lymph nodes, axillary lymph nodes, inguinal lymph nodes, gynecomastia, sympathetic ganglia, and any combination thereof.

38. A method of determining the suitability of surgical removal of cancer tissue in a subject having NTSR1 -positive cancer, or a subject at risk of having or developing NTSR1 -positive cancer, or a subject suspected of having or developing NTSR1 -positive cancer, comprising: (a) performing a PET scan on a subject using the ligand, PET probe or composition of any one of claims 1 to 23 or 26, wherein the PET scan identifies the presence of NTSR1 -positive cancer tissue; and (b) identifying the presence of NTSR1 -positive cancer tissue, wherein the presence of NTSR1 -positive cancer tissue indicates that the subject is suitable for surgical removal of the cancer tissue.

39. A method of treating an NTSR1 -positive cancer in a subject, comprising determining the suitability of a subject having an NTSR1 -positive cancer, or a subject at risk of having or developing an NTSR1 -positive cancer, or suspected of having or developing an NTSR1 -positive cancer, for surgical removal of cancer tissue by performing a PET scan on the subject using the ligand, PET probe or composition of any one of claims 1 to 23 or 26, wherein the PET scan identifies the presence of NTSR1 -positive cancer tissue, and treating the NTSR1 -positive cancer based on the results of the PET scan.

40. The method of claim 39, wherein treating an NTSR1 -positive cancer comprises administering the ligand, therapeutic agent or composition of any one of claims 1 to 21, 24 to 25 or 27.

41. A method of treating a disorder of NTSR1 -positive tissue in a subject, comprising determining the suitability of a subject having the disorder, or a subject at risk of having or developing the disorder, or suspected of having or developing the disorder, for treatment thereof by performing a PET scan on the subject using the ligand, PET probe or composition of any one of claims 1 to 23 or 26, wherein the PET scan identifies the presence of NTSR1 -positive tissue, and treating the disorder based on the results of the PET scan.

42. The method of claim 41, wherein treating the condition comprises administering the ligand, therapeutic agent or composition of any one of claims 1 to 21, 24 to 25 or 27.

43. The method of claim 41 or 42, wherein the condition is a cancer from a tissue selected from the group consisting of the prostate, liver, kidney, spleen, bladder, parotid gland, lacrimal gland, submandibular gland, small intestine, nasal mucosa, esophageal mucosa, vocal cords, gallbladder, bile duct, trachea, lung, breast, mediastinal lymph nodes, axillary lymph nodes, inguinal lymph nodes, gynecomastia, sympathetic ganglia, and any combination thereof.

44. The method of any one of claims 36 to 43, wherein the subject is a preoperative subject.

45. The method of any one of claims 36 to 43, wherein the subject is an intraoperative subject (eg, wherein the subject is undergoing surgery).

46. ​​The method of any one of claims 36 to 45, wherein treatment comprises surgical removal of at least a portion of the identified NTSR1 -positive tissue and / or administration of an anti-cancer therapeutic (e.g., a chemotherapeutic agent, a radiotherapeutic agent (e.g., a ligand, therapeutic agent, or composition of any one of claims 1 to 21, 24 to 25, or 27), an immunotherapeutic agent, or any combination thereof).

47. The method of any one of claims 30 to 34, 36, 38, 39 or 41, wherein performing a PET scan on the subject using the ligand, PET probe and / or composition of any one of claims 1 to 23 or 26 comprises administering about 1 to about 15 mCi of the ligand, PET probe and / or composition.

48. The method of claim 47, wherein administration is by intravenous injection.

49. The method of any one of claims 30 to 34, 36, 38, 39, 41 or 43 to 48, wherein the ligand, PET probe and / or composition has a tumor to muscle ratio (T / M) of at least 3 or more at about 1 hour after injection, optionally, a tumor to muscle ratio (T / M) of at least 10 or more at about 1 hour after injection.

50. The method of any one of claims 30 to 34, 36, 38, 39, 41 or 43 to 49, wherein the ligand, probe, therapeutic agent and / or composition renders a major organ transparent in 3 hours or less.

51. The method of claim 50, wherein the major organs include one or more organs selected from the group consisting of liver, kidney, muscle, and any combination thereof.

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