DLL3-targeted peptides and their constructs

Abstract

This disclosure relates to targeting moieties, such as peptides and antibodies, that can bind to DLL3. The disclosure also provides targeted constructs, which may include a targeting moiety conjugated to a chelating agent for cargo association via an optional linker. Methods for constructing the constructs and formulations thereof are also provided. Methods for using the constructs and / or formulations to treat a target, for example, to treat or prevent cancer, are also described.

Description

[Technical Field]

[0001] Related applications This application relates to U.S. Provisional Patent Application No. 63 / 508,191, filed on 14 June 2023, and U.S. Provisional Patent Application No. 63 / 557,132, filed on 23 February 2024. The entire contents of each application are incorporated herein by reference. [Background technology]

[0002] Radiotherapy, or radiation therapy, is a cancer treatment that uses high doses of radiation to kill cancer cells and shrink tumors. In recent years, targeted radionuclide therapy using radiolabeled peptides has been developed as an alternative to external beam radiation therapy. These peptides typically bind to receptors that are overexpressed by cancer cells. Despite these advances, the need for novel targeted radionuclide therapies remains.

[0003] Delta-like ligand 3 (DLL3), a member of the Notch signaling pathway, is a potential target for radionuclide therapy. This evolutionarily conserved system regulates cell fate through intercellular interactions. During embryonic development, DLL3 is highly expressed and transported to the cell membrane. Once development is complete, DLL3 expression becomes downregulated and confined to cells, typically within the Golgi apparatus. However, DLL3 expression has been found to be highly expressed and localized to the cell membrane in many forms of cancer (Xiu et al., Onco.Targets Ther. (2020), 13:3881-3901).

[0004] In addition to being a biomarker, DLL3 plays a role in regulating cancer behavior. Studies in small cell lung cancer (SCLC) have shown that upregulation of DLL3 expression reduces tumor sensitivity to chemotherapy. Furthermore, blocking DLL3 inhibited the proliferation and migration of SCLC cells and restored epithelial-mesenchymal transition (EMT) (Huang et al., Biochem. Biophys. Res. Commun. (2019) 514(3):853-860). The oncogenic behavior of DLL3 has also been demonstrated in pancreatic cancer, melanoma, and gastric cancer (Mullendore, et al., Clin. Cancer. Res. (2009) 15(7): 2291-301; Ding, et al., Life Sci. (2019) 226: 149-155; Hu et al., Nan Fang Yi Ke Da Xue Xue Bao. (2018) 38(1): 14-19).

[0005] In summary, these findings suggest that DLL3 plays a crucial role in regulating oncogenic pathways and is specifically upregulated in cancer cells; therefore, the development of therapies targeting DLL3 would be beneficial for the clinical treatment of cancer. [Overview of the project]

[0006] This disclosure relates to targeting moieties such as peptides, proteins, and antibodies that can bind to delta-like canonical Notch ligand 3 (DLL3). The disclosure also provides targeting constructs which may include a targeting moiety conjugated to a chelating agent for cargo association via an optional linker. The chelating agent may associate with a payload such as, for example, a radionuclide or a cytotoxic agent.

[0007] In certain embodiments, the targeted construct comprises a targeted moiety, which is a cyclic peptide targeting DLL3, which is conjugated via an optional linker to a chelating agent for the association of a radioisotope or radionuclide.

[0008] Therefore, provided herein are cyclic peptides that target DLL3. Accordingly, these peptides are useful for the treatment of various indications including cancer.

[0009] In one aspect, provided herein is a cyclic peptide comprising the amino acid sequence of Formula A:

Chemical formula

[0010] In one aspect, provided herein is a cyclic peptide of Formula I:

Chemical formula

[0011] In one embodiment, the cyclic peptide of Formula I is attached via an optional linker to a chelator for labeling with a radionuclide.

[0012] In another embodiment, the cyclic peptide of Formula I is selected from the cyclic peptides in Table A. In yet another embodiment, the cyclic peptide of Formula I is selected from the cyclic peptides in Table B.

[0013] In yet another embodiment, the cyclic peptide of Formula I is selected from the cyclic peptides in Table A or a pharmaceutically acceptable salt and / or solvate thereof. In yet another embodiment, the cyclic peptide of Formula I is selected from the cyclic peptides in Table B or a pharmaceutically acceptable salt and / or solvate thereof.

[0014] In another aspect, provided herein is a cyclic peptide of Formula B: [Chemical formula] or a pharmaceutically acceptable salt thereof, wherein the variables are defined herein.

[0015] In one embodiment, the cyclic peptide of Formula B is attached to a chelating agent via an optional linker, and the chelating agent is optionally labeled with a radionuclide.

[0016] In another embodiment, the cyclic peptide of Formula B is selected from the cyclic peptides in Table A. In yet another embodiment, the cyclic peptide of Formula B is selected from the cyclic peptides in Table B.

[0017] In another embodiment, the cyclic peptide of Formula B is selected from the cyclic peptides in Table A or a pharmaceutically acceptable salt and / or solvate thereof. In yet another embodiment, the cyclic peptide of Formula B is selected from the cyclic peptides in Table B or a pharmaceutically acceptable salt and / or solvate thereof.

[0018] In yet another embodiment, the chelating agent is selected from the chelating agents in Table C. In some embodiments, the chelating agent is labeled with a radionuclide. In some embodiments, the radionuclide is 111 In, 99m Tc, 94m Tc, 66 Ga, 67 Ga, 68 Ga, 52 Fe, 169 Er, 72 As, 97 Ru, 203 Pb, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 89 Sr, 186 Re, 188 Re, 86 Y, 90 Y, 89 Zr,51 Cr, 52 Mn, 51 Mn, 177 Lu, 169 Yb, 175 Yb, 105 Rh, 166 Dy, 166 Dy, 166 Ho, 153 Sm, 149 PM, 151 PM, 172 Tm, 121 Sn, 117 mSn, 212 Bi, 213 Bi, 142 Pr, 143 Pr, 198 Au, 199 Au, 123 I, 124 I, 125 I, 131 I, 75 Br, 76 Br, 77 Br, 80 Br, 82 Br, 18 F, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 43 Sc, 44 Sc, 47 Sc, 212 Pb, 211 At, 223 Ra, 227 Th, 226 Th, 82 Rb, 32 P, 76 As, 89 Zr, 111 Ag, 165 Er, 225 Ac, and 227 Selected from the group consisting of Ac. In some embodiments, the radionuclide is 111 In, 99m Tc, 67 Ga, 68 Ga, 203 Pb, 64 Cu, 86 Y, 89 Zr, 123 I,124 I、 125 I、 18 F、 76 Br、 77 Br、 152 Tb、 155 Tb、 44 Sc、 43 Sc、 67 Cu、 188 Re、 90 Y、 177 Lu、 213 Bi、 131 I、 47 Sc、 225 Ac、 212 Pb、 211 At, or 227 Th. [[ID=​​​​​​​​​​​​​In yet another embodiment, the foregoing provides a method for treating cancer in a target subject of interest, comprising administering a therapeutically effective amount of one of the compounds described herein to the target subject. The cancer may comprise at least one cell containing DLL3. The cancer may be urothelial carcinoma, melanoma, or squamous cell carcinoma. Furthermore, the cancer may be a neuroendocrine neoplasm, melanoma, or primary brain tumor. In some embodiments, the neuroendocrine tumor is selected from small cell lung cancer (SCLC), medullary thyroid carcinoma (MTC), large cell neuroendocrine carcinoma (LCNEC), pancreatic and gastrointestinal neuroendocrine carcinoma (GEP NEC), neuroendocrine prostate cancer (NEPC), small cell prostate cancer (SCPC), Merkel cell carcinoma (MCC), neuroendocrine cervical cancer, grade 3 neuroendocrine tumor (NET), and extrapulmonary neuroendocrine carcinoma of the cervix (NEC). In some embodiments, the cancer is a solid tumor that is DLL3-positive (e.g., ≥1% DLL3-positive cells) as measured by immunohistochemistry (IHC).

[0023] In one embodiment, the herein provides a peptide having binding specificity to DLL3, which binds to one or more amino acids A81, L83, G106, A85, and R61 of the DLL3 amino acid sequence of SEQ ID NO: 1. In one embodiment, the peptide binds to amino acids A81, L83, G106, A85, and R61 of the DLL3 amino acid sequence of SEQ ID NO: 1. In another embodiment, the peptide binds to the main chain atoms of amino acids A81, L83, G106, and A85 of the DLL3 amino acid sequence of SEQ ID NO: 1. In yet another embodiment, the peptide binds to the side chain atom of amino acid R61 of the DLL3 amino acid sequence of SEQ ID NO: 1. In yet another embodiment, the peptide binds to the main chain atoms of amino acids A81, L83, G106, and A85 of the DLL3 amino acid sequence of SEQ ID NO: 1, and to the side chain atom of amino acid R61 of the DLL3 amino acid sequence of SEQ ID NO: 1. In one embodiment, the peptide includes the amino acid sequence of WTACANAKDCWP, or a derivative thereof containing one or more non-natural amino acids. In another embodiment, amino acids W1, A3, A7, and W11 are bound to DLL3. In yet another embodiment, the peptide is cyclic. In yet another embodiment, the peptide is acyclic.

[0024] In some embodiments, the Disclosure provides a construct comprising a targeted moiety conjugated to at least one chelating agent for cargo association via an optional linker, or a pharmaceutically acceptable salt thereof, wherein the targeted moiety conjugates to a cellular antigen, and the cellular antigen comprises DLL3. The cargo may be a payload such as a radionuclide or a cytotoxic agent. The targeted moiety may comprise a peptide. The chelating agent may comprise a polyaminocarboxylate. The chelating agent may comprise ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 1,4,7,10-tetra-azacyclododecane-N,N',N”,N”'-tetraacetic acid (DOTA), DOTAGA, or derivatives thereof. The chelating agent may include EDTA, DTPA, DOTA, DOTAGA, or derivatives thereof. The chelating agent may include macrocyclic agents. The chelating agent may be 1,4,7-triazacyclononane-N,N',N”-triacetic acid (NOTA), 1,4,7,10-tetraazacyclododecane-N,N',N”,N”'-tetraacetic acid (TETA), 1,4,7,10,13-pentazacyclopentadecane-N,N',N”,N”',N””-pentaacetic acid (PEPA), 1,4,7,10,13,16-hexaazacyclohexadecane-N,N',N”,N”',N””,N””'-hexaacetic acid (HEHA), or These may include their derivatives. The chelating agents may also include deferoxamine (DFO), 5,11,16,22-tetraazahexacosanediamide (DFO*), or N,N'-1,4-butanediylbis[N-[3-[[(1,6-dihydro-1-hydroxy-6-oxo-2-pyridinyl)carbonyl]amino]propyl]-1,6-dihydro-1-hydroxy-6-oxo-2-pyridinecarboxamide](HOPO), or their derivatives.

[0025] The cargo may contain a radioactive agent. The radioactive agent may contain a radioisotope. Therefore, in some embodiments, the constructs or compounds disclosed herein may optionally contain a radioisotope. In some embodiments, the constructs or compounds disclosed herein contain a radioisotope. A radioisotope may be a radionuclide. A radioisotope may be any of those listed in Table 3. In some embodiments, the radionuclide is 111 In, 99m Tc, 94m Tc, 66 Ga, 67 Ga, 68 Ga, 52 Fe, 169 Er, 72 As, 97 Ru, 203 Pb, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 89 Sr, 186 Re, 188 Re, 86 Y, 90 Y, 89 Zr, 51 Cr, 52 Mn, 51 Mn, 177 Lu, 169 Yb, 175 Yb, 105 Rh, 166 Dy, 166 Dy, 166 Ho, 153 Sm, 149 PM, 151 PM, 172 Tm, 121 Sn, 117 mSn, 212 Bi, 213 Bi, 142 Pr, 143 Pr, 198 Au, 199 Au, 123 I, 124 I, 125 I, 131 I, 75 Br, 76 Br, 77 Br, 80Br, 82 Br, 18 F, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 43 Sc, 44 Sc, 47 Sc, 212 Pb, 211 At, 223 Ra, 227 Th, 226 Th, 82 Rb, 32 P, 76 As, 89 Zr, 111 Ag, 165 Er, 225 Ac, and 227 Selected from the group consisting of Ac. In some embodiments, the radionuclide is 111 In, 99m Tc, 67 Ga, 68 Ga, 203 Pb, 64 Cu, 86 Y, 89 Zr, 123 I, 124 I, 125 I, 18 F, 76 Br, 77 Br, 152 Tb, 155 Tb, 44 Sc, 43 Sc, 67 Cu, 188 Re, 90 Y, 177 Lu, 213 Bi, 131 I, 47 Sc, 225 Ac, 212 Pb, 211 At, or 227 It is Th. An optional linker may include a cleavable linker. An optional linker may include an incleavable linker. An optional linker may include at least one amino acid.

[0026] In some embodiments, the disclosure provides pharmaceutical compositions comprising a construct and pharmaceutically acceptable excipients.

[0027] In some embodiments, the disclosure provides a method for delivering a cargo to cells, comprising contacting cells or a cell-containing object with a construct or a pharmaceutical composition thereof. In one embodiment, the cargo may be a radioagent such as a radionuclide and / or radioisotope. In other embodiments, the cargo may be a cytotoxic agent.

[0028] In some embodiments, the Disclosure provides a method for treating a disease or disorder of interest, comprising administering a construct or a pharmaceutical composition thereof. In some embodiments, the Disclosure provides a method for treating a subject, comprising administering a construct or a pharmaceutical composition thereof. In some embodiments, the disease or disorder is cancer (i.e., the subject has cancer). The cancer may comprise at least one cell containing DLL3. In one embodiment, the cancer expresses DLL3. The cancer may be urothelial carcinoma, melanoma, or squamous cell carcinoma. Furthermore, the cancer may be a neuroendocrine neoplasm, melanoma, or primary brain tumor. In some embodiments, the neuroendocrine tumor is selected from small cell lung cancer (SCLC), medullary thyroid carcinoma (MTC), large cell neuroendocrine carcinoma (LCNEC), pancreatic and gastrointestinal neuroendocrine carcinoma (GEP NEC), neuroendocrine prostate cancer (NEPC), small cell prostate cancer (SCPC), Merkel cell carcinoma (MCC), neuroendocrine cervical cancer, and grade 3 neuroendocrine tumors (NETs). In some embodiments, the cancer is a solid tumor that is DLL3-positive (e.g., ≥1% DLL3-positive cells) as measured by immunohistochemistry (IHC). [Modes for carrying out the invention]

[0029] DLL3 regulates embryonic development by binding to members of the highly conserved Notch receptor family. In contrast to canonical Notch ligands, DLL3 represses Notch signaling through interaction with the Golgi apparatus. Reflecting this function, DLL3 is typically confined to the cytoplasm (Geffers, I., et al. J. Cell Biol. (2007) 178 76(3), 465; Zhou, B., et al. Signal Transduct. Target Ther. (2022) 7(1) Signal Transduct. 95). DLL3 is detected in the cytoplasm of healthy fetal tissue, and its absence results in severe vertebral defects in the form of autosomal recessive spondylocostal dysplasia (Serth, K., et al. PLoS One, (2015) 10(4), e0123776; Dunwoodie, SL, et al. Development, (2002) 129(7), 1795-806). Low levels of DLL3 are detectable as RNA transcripts in the adult brain, pituitary gland, and testes (Sharma, SK, et al. Cancer Res. (2017) 77(14), 3931-41). DLL3 mRNA is also detectable in the cytoplasm of the adult brain, pituitary gland, basophils, and pancreas (Giffin, MJ, et al. Clin. Cancer Res, (2021) 27(5), 1526-37). However, significant overexpression of DLL3 leads to its abnormal localization to the cell surface (Zhou, B., et al. Signal Transduct. Target Ther. (2022) 7(1), 95; Geffers, I., et al. (2007)). Upregulated DLL3 has also been observed to abnormally localize on the cell surface in cancer (Giffin, MJ, et al. (2021); Saunders, LR, et al. Sci. Transl. Med. (2015) 7(302), 302ra136; Sharma, SK, et al. (2017)).

[0030] Abnormal DLL3 expression, including cell surface expression, is observed in various human tumors (Saunders, LR, et al. (2015)). Neuroendocrine neoplasms (NENs), including well-differentiated neuroendocrine neoplasms (NETs) and poorly differentiated neuroendocrine carcinomas (NECs), often express DLL3 on the cell surface and share common histological and transcriptome markers for the neuroendocrine system and transformation (Puca, L., et al. Sci. Transl. Med. (2019) 11(484); Yao, J., et al. Oncologist (2022) 27(11), 940-51). Although the pathophysiological role of abnormal localization of DLL3 on tumor cell function is not fully understood, acquisition and deletion experiments suggest that DLL3 may influence cell proliferation, migration, and tumor growth in vitro and in vivo (Furuta, M., et al. Cancer Sci. (2019) 110(5), 1599-608; Huang, J., et al. (2019). DLL3 is detectable on the tumor cell surface by immunohistochemistry (IHC) and flow cytometry, and exogenous DLL3-targeting antibodies are accessible in vivo (Dylla, SJMol. Cell Oncol. (2016) 3(2), e1101515; Saunders, LR, et al. (2015)).

[0031] DLL3 has been quantified at a low single-order-of-magnitude, several thousand copies per cell in representative small cell lung cancer (SCLC) and neuroendocrine prostate cancer (NEPC) cell lines (Giffin, MJ, et al. (2021); Zhang, Y., et al. Clin. Cancer Res. (2023) 29(5), 971-85). Despite the low copy number levels per cell, the feasibility of targeting DLL3 has nevertheless been demonstrated preclinically in SCLC and neuroendocrine carcinoma xenograft models using DLL3-targeted 89Zr / 177Lu radioconjugates, DLL3-targeted bispecific compounds, and DLL3-targeted antibody-drug conjugates (Chou, J., et al. Cancer Res. (2023) 83(2), 301-15; Giffin, MJ, et al. (2021); Korsen, JA, et al. Proc. Natl. Acad. Sci. USA, (2022) 119(27), e2203820119; Saunders, LR, et al. (2015)). Furthermore, despite the low abundance of tumor-associated antigens, visualization of DLL3-expressing tumors by 89Zr immunoPET imaging has been demonstrated in several mouse models of SCLC (Sharma, SK, et al. (2017)).

[0032] Its clinical validation as a target has been confirmed with DLL3-targeted therapies, including the DLL3-targeted antibody-drug conjugate RovaT (Morgensztern, D., et al. Clin Cancer Res, (2019) 25(23), 6958-66; Rudin, CM, et al. Nat Rev Dis Primers (2021) 7(1), 3), although its potential was limited due to payload-related toxicity. More recently, DLL3-targeted bispecific tarlatamab (AMG757) has resulted in sustained responses of up to 12 months in a quarter of treated patients (Paz-Ares, L., et al. J. Clin. Oncol. (2023) 41(16), 2893-903). Given its well-studied nature and its selective expression on the surface of cancer cells in both primary and metastatic solid tumors, DLL3 is a compelling target for novel therapies in NENs and other solid tumors expressing DLL3.

[0033] Provided herein are compounds that target DLL3. In particular, provided herein are targeted constructs (also referred to herein as “compounds”) comprising a targeting moiety, which is a cyclic peptide targeting DLL3, conjugated to a chelating agent for the association of a radioisotope via an optional linker. Thus, these compounds, as well as pharmaceutical compositions comprising these compounds, are useful for the treatment of a variety of indications, including cancer.

[0034] I. Compounds and Compositions In some embodiments, this disclosure relates to targeting moieties, such as peptides, proteins, and antibodies, that can bind to a target. In some embodiments, this disclosure provides constructs that can localize to a target and / or associate with a target. Such constructs, including any combination of targeting moieties and cargo, are referred to herein as “targeting constructs.” As used herein, the term “targeting moiety” refers to a component or combination of components of a targeting construct that is involved in the targeting of the localization of the construct to a target or the association with a target. Cargo components of a targeting construct may include, but are not limited to, any one of a variety of compounds, including compounds, biomolecules, metals, polymer molecules, therapeutic agents, cytotoxic agents, and radioactive agents. Chelating agents may associate with payloads, for example, radionuclides or cytotoxic agents.

[0035] In particular, provided herein are targeted constructs (also referred to herein as “compounds”) comprising a targeted moiety, which is a cyclic peptide targeting DLL3, conjugated to a chelating agent for the association of radioisotopes via an optional linker.

[0036] target Targeted constructs can be directed to a variety of targets. In certain embodiments, a targeted construct includes a peptide directed to DLL3 and also includes a radioisotope. In some embodiments, a targeted construct can target cells. Such a targeted construct may include a targeting moiety that can target cellular antigens, including those that associate with the surface of a target cell. In this case, the cellular antigen is the target of the targeting moiety and the targeted construct. Where used herein, “antigen” may refer to any entity that induces an immune response in an organism, or simply an antibody-binding partner. An immune response is the reaction of a cell, tissue, and / or organ of an organism to a foreign substance. An immune response typically results in the production of one or more antibodies against the foreign substance by the organism. Where used herein, the term “target antigen” refers to an entity, protein, or epitope to which an antibody binds, or to which an antibody is desired, designed, or developed to have affinity. Such target antigens may include cancer cell antigens, for example, those expressed on the surface of cancer cells.

[0037] In some embodiments, the target antigen of this disclosure comprises DLL3 or a portion thereof. The DLL3 antigen may comprise the extracellular domain of DLL3. The DLL3 antigen may comprise a fusion protein of DLL3 or other entities comprising a DLL3 portion.

[0038] DLL3 Delta-like canonical Notch ligand 3 (also known as delta-like protein 3, Drosophila, or DLL3) is a member of the delta protein ligand family. It is encoded by the DLL3 gene. DLL3 is involved in inhibiting primary neurogenesis and diverting neurons along specific differentiation pathways. It also plays a role in the formation of somatic cell boundaries during the segmentation of the paraxial mesoderm. Mutations in the DLL3 gene cause Yarcho-Levin syndrome, an autosomal recessive genetic disorder. Expression of the DLL3 gene occurs in neuroendocrine tumors. DLL3 may be a promising target for treating tumors such as lung cancer.

[0039] In some embodiments, the targeted construct includes a targeting moiety specific to one or more DLL3 domains. In some embodiments, the targeted construct includes a targeting moiety specific to one or more DLL3 target amino acid sequences or fragments or variants thereof listed in Table 1.

[0040] [Table 1]

[0041] targeting part In some embodiments, the targeting moiety localizes the targeting construct to a target by binding to such target or related components. The targeting moiety may bind to cells or biomolecules or other structures related to cells. For example, in some embodiments, the targeting moiety binds to cellular antigens. Such cellular antigens may be specifically expressed by, on, or otherwise associated with a particular cell type. A particular cell type may be characterized by one or more of the following: cell size, age, shape, location, tissue of origin, organ of origin, function, activity, genotype, phenotype, or association with dysfunction or disease. The targeting moiety may bind to cancer cell antigens. In some embodiments, the targeting moiety binds to DLL3. In some embodiments, the targeting moiety binds to human DLL3.

[0042] The targeted moiety may include or consist of a protein, polypeptide, antibody, nucleic acid, nucleic acid analog, aptamer, lipid, carbohydrate, glycoprotein, or small molecule. In some embodiments, the targeted moiety may include or consist of a polypeptide or peptide, an antibody or a fragment or variant thereof. In certain embodiments, the targeted moiety is a cyclic peptide.

[0043] In some embodiments, the targeted portions of this disclosure, such as peptides and antibodies, have affinity for human DLL3. In some embodiments, the targeted portions of this disclosure have affinity for human DLL3 within an identified range when measured by conventional assays. "Affinity" or "binding affinity" means the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody or peptide-bonded compound) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects all interactions between the members of a binding pair (e.g., an antibody or peptide-bonded compound and an antigen). The affinity of molecule X for its partner Y is generally expressed by the dissociation constant (K D It can be expressed by the equilibrium dissociation constant (K). D ) is calculated as the koff / kon ratio.

[0044] Low-affinity targeting moieties generally tend to bind slowly to antigens and dissociate easily, while high-affinity targeting moieties generally tend to bind more rapidly to antigens and remain bound for a longer period. Various methods for measuring binding affinity are known in the art, and any of them can be used for the purposes of this disclosure.

[0045] In some embodiments, the targeted portion disclosed herein has an equilibrium dissociation constant (K) of approximately 0.001 nM to approximately 0.01 nM, approximately 0.005 nM to approximately 0.05 nM, approximately 0.01 nM to approximately 0.1 nM, approximately 0.05 nM to approximately 0.5 nM, approximately 0.1 nM to approximately 1.0 nM, approximately 0.5 nM to approximately 5.0 nM, approximately 2 nM to approximately 10 nM, approximately 8 nM to approximately 20 nM, approximately 15 nM to approximately 45 nM, approximately 30 nM to approximately 60 nM, approximately 40 nM to approximately 80 nM, approximately 50 nM to approximately 100 nM, approximately 75 nM to approximately 150 nM, approximately 100 nM to approximately 500 nM, approximately 200 nM to approximately 800 nM, approximately 400 nM to approximately 1,000 nM, or at least 1,000 nM. DIt can bind to the target protein at a constant (K) of about 1 nM or less, about 1 nM to about 10 nM, about 10 nM to about 100 nM, or about 100 nM to about 300 nM. D ) can bind to the target protein. In some embodiments, the target protein is DLL3.

[0046] In some embodiments, K D This is determined by surface plasmon resonance (SPR). An exemplary SPR protocol is provided in Example 3.

[0047] polypeptides and peptides In some embodiments, the targeted portion of this disclosure is a polypeptide. According to this disclosure, any amino acid-based molecule (natural or unnatural) may be referred to as a “polypeptide,” a term that encompasses “peptides,” “peptide mimes,” and “proteins.” A “peptide” is conventionally considered to be in the size range of about 4 to about 50 amino acids. Peptides larger than about 50 amino acids are generally referred to as “proteins.”

[0048] The peptides of this disclosure may be linear or cyclic. In particular, cyclic peptides targeting DLL3 are provided herein. Cyclic peptides include any peptide having one or more cyclic features as part of its structure, such as loops and / or internal bonds. In some embodiments, cyclic peptides are formed when a molecule acts as a crosslinking portion connecting two or more regions of a peptide.

[0049] As used herein, the term “crosslinked portion” refers to one or more components of a crosslink formed between two adjacent or non-adjacent amino acids, unnatural amino acids, or non-amino acids in a peptide. A crosslinked portion may be of any size or composition. In some embodiments, a crosslinked portion may include one or more chemical bonds between two adjacent or non-adjacent amino acids, unnatural amino acids, non-amino acid residues, or combinations thereof. In some embodiments, such chemical bonds may be between one or more functional groups on adjacent or non-adjacent amino acids, unnatural amino acids, non-amino acid residues, or combinations thereof. A crosslinked portion may include one or more of the following: amide bonds (lactams), disulfide bonds, thioether bonds, aromatic rings, triazole rings, and hydrocarbon chains. In some embodiments, a crosslinked portion includes an amide bond between an amine functional group and a carboxylate functional group, which are each present in the side chain of an amino acid, unnatural amino acid, or non-amino acid residue. In some embodiments, the amine or carboxylate functional group is part of a non-amino acid residue or an unnatural amino acid residue.

[0050] In some embodiments, the Disclosure provides peptides that bind to human DLL3. Antibodies of the Disclosure may bind to the extracellular domain of human DLL3. Antibodies of the Disclosure may bind to DLL3 associated with cells (e.g., on the cell surface). Such cells include, but are not limited to, cancer cells such as lung cancer cells, breast cancer cells, bladder cancer cells, colon cancer cells, urothelial carcinoma cells, melanoma cells, or squamous cell carcinoma cells.

[0051] DLL3 epitope The peptides of this disclosure that bind to human DLL3 bind to specific amino acids in DLL3, i.e., DLL3 epitopes. As used herein, the term “epitope” refers to a specific portion of a target (e.g., DLL3) that interacts with (e.g., binds to) a binding entity (e.g., the peptides of this disclosure).

[0052] In some embodiments, the peptide has binding specificity to DLL3, where the peptide binds to one or more amino acids A81, L83, G106, A85, and R61 of the DLL3 amino acid sequence of SEQ ID NO: 1.

[0053] In some embodiments, the peptide binds to amino acids A81, L83, G106, A85 and R61 of the DLL3 amino acid sequence of SEQ ID NO: 1.

[0054] In some embodiments, the peptide is bonded to the main chain atoms of amino acids A81, L83, G106, and A85 of the DLL3 amino acid sequence of SEQ ID NO: 1. As used herein, the term “main chain atom” refers to an atom in the peptide backbone, which consists of a central carbon atom (α-carbon) bonded to a hydrogen atom, an amino group (NH2), and a carboxyl group (COOH).

[0055] In some embodiments, the peptide is bound to the side chain atom of amino acid R61 in the DLL3 amino acid sequence of SEQ ID NO: 1. As used herein, the term “side chain atom” refers to an atom in the peptide side chain, also commonly referred to as an R group, and in some embodiments, the peptide is bound to the main chain atoms of amino acids A81, L83, G106 and A85 in the DLL3 amino acid sequence of SEQ ID NO: 1, and to the side chain atom of amino acid R61 in the DLL3 amino acid sequence of SEQ ID NO: 1.

[0056] In another embodiment, the peptide is approximately 1 × 10 -8 M ~ approx. 1×10 -12 M EC 50 It can be bound to DLL3 by value. In another embodiment, the peptide is approximately 1 × 10 -8 M ~ approx. 1×10 -10 M EC 50 It can be bound to DLL3 by value.

[0057] In some embodiments, EC 50The value is determined by enzyme-linked immunosorbent assay (ELISA). In some embodiments, DLL3 protein concentrations of approximately 1 μg / mL to approximately 5 μg / mL are used in ELISA.

[0058] In some embodiments, the peptide comprises the amino acid sequence WTACANAKDCWP, or a derivative thereof containing one or more non-natural amino acids. In another embodiment, amino acids W1, A3, A7, and W11 are bound to DLL3. In yet another embodiment, the peptide is cyclic. In yet another embodiment, the peptide is acyclic. In one embodiment, the peptide is any of the formulas or species described herein.

[0059] Cyclic peptide In some embodiments, the peptides of this disclosure may comprise cyclic peptides having one or more crosslinking sites (e.g., cyclic structures, staples, crosslinks, etc.). Peptide stapling / crosslinking is a macrocyclic modification technique in which a peptide is covalently modified by the formation of chemical bonds (e.g., staples, crosslinking sites, etc.) between the side chains of two amino acids. More specifically, a peptide is made macrocyclic by the formation of covalent bonds between atoms present in the linear peptide and atoms of the crosslinking site. Stapling / crosslinking can be used to constrain a peptide to a preferred bioactive conformation (reducing conformational flexibility and rotational freedom), thereby improving affinity to specific receptor targets and improving overall pharmacokinetics. The linked residues are generally located on the same plane of the peptide helix and separated by one, two, or three helix turns (e.g., the first amino acid at position (z) is linked to a second amino acid at positions z+4, z+7, or z+11). In some embodiments, the crosslinked portion may contain one or more chemical bonds between two adjacent or non-adjacent amino acids, unnatural amino acids, non-amino acid residues, or combinations thereof. In some embodiments, such chemical bonds may be between one or more functional groups on adjacent or non-adjacent amino acids, unnatural amino acids, non-amino acid residues, or combinations thereof.

[0060] Therefore, in one embodiment, the herein provides a cyclic peptide comprising the amino acid sequence of formula A: [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, X0 is any natural or non-natural amino acid, or X0 is absent; X1 is selected from Trp, 7aza-Trp, 1Me-Trp, 5OH-Trp, 5Ome-Trp, 7Ome-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, α-Me-Trp, and NMe-Trp; X2 and X3 are each independently any natural or non-natural amino acid; Y1 is Cys; X4, X5, X6, X7, and X8 are each independently any natural or non-natural amino acid; Y2 is Cys; X9 is selected from Trp, 7aza-Trp, 1Me-Trp, 5OH-Trp, 5Ome-Trp, 7Ome-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, α-Me-Trp, and NMe-Trp; X 10 The following are selected from Pro, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, 5,5-di-Me-Pro, trans4NH2-Pro, and cis4NH2-Pro; P 1 is, -L 1 -Chelating agents, [ka] Selected from; D 1 It is a -NR”-chelating agent; L 1 It does not exist, or [ka] Selected from; Here, L 1 The amino group is P 1 Alternatively, it can bond to the carbonyl group of the chelating agent to form an amide bond; Each n, q, and u is an integer between 0 and 16, independently of the others; Each p is an integer between 0 and 24, independently; Each s is an independent integer between 0 and 16; Each t is independently 1, 2, 3, 4, 5, or 6; R' is independently selected from H, C(O)OH, (CH2)OH, and NHAc; R is independently selected from H and CH3; Here, the cyclic peptide is cyclized via a linker between Y1 and Y2; Here, the cyclic peptide binds to DLL3.

[0061] In one embodiment, X2 is Thr and X8 is Asp.

[0062] In another embodiment, the cyclic peptide of formula A is a cyclic peptide comprising the amino acid sequence of formula Ai: [ka] or a pharmaceutically acceptable salt thereof, In the formula, R'' is independently selected from H and CH3.

[0063] In yet another embodiment, the cyclic peptide of formula A is a cyclic peptide comprising the amino acid sequence of formula Aii: [ka] or a pharmaceutically acceptable salt thereof, In the formula, R'' is independently selected from H and CH3.

[0064] In yet another embodiment, the cyclic peptide of formula A is a cyclic peptide comprising the amino acid sequence of formula Aiii: [ka] or a pharmaceutically acceptable salt thereof, In the formula, R'' is independently selected from H and CH3.

[0065] In one embodiment, the cyclic peptide of formula B is a cyclic peptide comprising the amino acid sequence of formula Biii: [ka] In the formula, R is independently selected from H, CH3, OH, halogen, C(O)OH, and N(R)2.

[0066] In another embodiment, the C-terminus of the cyclic peptide is capped. In yet another embodiment, X 10 The C-terminus of the adjacent cyclic peptide is P 2’ Capped with P 2’ NH2, OH, CH2OH, [ka] Selected from; D 2 It is either OH or NH2; L 2 It does not exist, or [ka] Selected from; Here, L 2 The amino group is P 2 It links to the carbonyl group to form an amide bond; P 3 H, Ac, [ka] Selected from; L 3 It does not exist, or independently, [ka] Selected from; Here, L 3 The carbonyl group is P 2 It links to the amine group to form an amide bond; D 3 These are independently CH3, C(O)OH, and [ka] Selected from; X is a halogen, Each n is an independent integer between 0 and 16; Each p is an integer between 0 and 24, independently; Each s is an independent integer between 0 and 16; Each t is independently 1, 2, 3, 4, 5, or 6; w is selected from 1, 2, or 3.

[0067] In another embodiment of the above configuration, X0 is absent, or X0 is selected from Gly, Met, D-Ala, Ala, Nle, and Nva. In yet another embodiment, X0 is absent.

[0068] In one embodiment of the above configuration, X1 is selected from Trp or 7Me-Trp. In another embodiment, X1 is Trp. If X0 is not present, X1 is P 1 It can be substituted with an N-terminal group selected from the following.

[0069] In one embodiment of the above-described model, X2 is selected from Thr, D-Ala, Ala, α-Me-Thr, Lys, and NMe-Thr. In another embodiment, X2 is selected from the group consisting of Thr, α-Me-Thr, and NMe-Thr. In yet another embodiment, X2 is Thr.

[0070] In yet another embodiment of the above-described portion, X3 is selected from Ile, Env, CHA, CBA, Nle, Tbg, THPG, Chg, 2Nal, 1Nal, 2CF3-Phe, 2PhEt-Ala, D-Ala, Ala, Leu, t-Bu-Ala, NMe-Nle, α-tert-amyl Gly, Allo-Ile, Lys(C12), Lys(C14), Lys(C16), α-Me-Ile, and NMe-tBuAla. In one embodiment, X3 is selected from the group consisting of Ile, Allo-Ile, α-Me-Ile, and NMe-tBuAla. In another embodiment, X3 is selected from the group consisting of D-Ala, Ala, t-Bu-Ala, and NMe-tBuAla. In yet another embodiment, X3 is Ile or NMe-tBuAla. In yet another embodiment, X3 is Ile. In one embodiment, X3 is NMe-tBuAla.

[0071] In another embodiment of the above configuration, the linker between Y1 and Y2 is connected, C 1~6 Alkylene, and [ka] Selected from.

[0072] In yet another embodiment of the above-described part, X4 is Asn, D-Ala, Ala, DAB-4-NHCOC5H 11 DAB-4-NHCOC7H 15 X4 is selected from Asp, Ser, Lys, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, 3Pya, 4Pya, Glu, and NMe-Asn. In yet another embodiment of the above aspects, X4 is selected from Asn, 3-(4-piperidinyl)-Ala, and Pip(CH2CO2H)Ala. In one embodiment, X4 is 3-(4-piperidinyl)-Ala or Pip(CH2CO2H)Ala. In another embodiment, X4 is Pip(CH2CO2X)Ala, where X is a pharmaceutically acceptable cation.

[0073] In another embodiment of the above-described aspect, X5 is selected from Asn, Ala, D-Ala, Trp, Asp, Lys, 3Pya, 4Pya, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, Glu, NMe-Asn, and Ser. In yet another embodiment of the above-described aspect, X5 is Asn or NMe-Asn. In yet another embodiment, X5 is Asn. In yet another embodiment, X5 is NMe-Asn.

[0074] In one embodiment of the above-described model, X6 is selected from Trp, 4CF3-Phe, 1Me-Trp, 7aza-Trp, BIP, 2Nal, 1Nal, aMe-Trp, D-Ala, Ala, 4F-Phe, 5F-Trp, 5Ome-Trp, Asn, 5OH-Trp, 7Me-Trp, 7MeO-Trp, 7Cl-Trp, and NMe-Trp. In another embodiment, X6 is selected from the group consisting of Trp, 1Me-Trp, 7aza-Trp, 2Nal, 1Nal, α-Me-Trp, 5F-Trp, 5MeO-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, 7Cl-Trp, and NMe-Trp. In yet another embodiment of the above-described model, X6 is selected from Trp, 2Nal, and 1Nal. In yet another embodiment, X6 is Trp. In one embodiment, X6 is 2Nal.

[0075] In another embodiment of the above-described aspect, X7 is selected from 3Pya, 4Pya, Lys(Me)3, His, Ala, D-Ala, Gln, Lys, Glu, Arg, Orn, NMe-His, and Ser. In yet another embodiment, X7 is His or Lys. In yet another embodiment, X7 is His. In one embodiment, X7 is Lys.

[0076] In another embodiment of the above-described model, X8 is selected from Asp, Asn, NMe-Asp, and α-Me-Asp. In yet another embodiment, X8 is Asp.

[0077] In yet another embodiment of the above-described part, X9 is Trp.

[0078] In one embodiment of the above-described configuration, X 10 It is Pro.

[0079] In another embodiment, the chelating agent is selected from the chelating agents in Table C. In yet another embodiment, the chelating agent is DOTA.

[0080] In another embodiment, the cyclic peptide of formula I is provided herein: [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, P 1 is, -L 1 -Chelating agents, [ka] Selected from; D 1 It is a -NR”-chelating agent; L 1 It does not exist, or [ka] Selected from; Here, L 1 The amino group is P 1 Alternatively, it can bond to the carbonyl group of the chelating agent to form an amide bond; P 2 C(O)NH2, C(O)OH, [ka] Selected from; D 2 It is either OH or NH2; L 2 It does not exist, or [ka] Selected from; Here, L 2 The amino group is P 2 It links to the carbonyl group to form an amide bond; P 3 H, [ka] Selected from; L 3 It does not exist, or independently, [ka] Selected from; Here, L 3 The carbonyl group is P 2 It links to the amine group to form an amide bond; D 3 These are independently CH3, C(O)OH, and [ka] Selected from; X is a halogen, R 0 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 1 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 2 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 3 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; B 1 C 1~6 It is alkylene; C 1 C 1~6 It is alkylene; A1 teeth, [ka] Selected from; Here, w is selected from 1, 2, or 3; R 4 This refers to the amino acid side chain of a natural amino acid or a non-natural amino acid, both of which are CH2C(O)OH or C(O)(CH2CH2O). p (CH2)2N(CH3)3 + It is arbitrarily replaced by; R 5 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 6 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 7 teeth, (i) amino acid side chains of natural amino acids, (ii) Selected from the amino acid side chains of non-natural amino acids, (iii) [ka] Selected from the group consisting of; R 8 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 9 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 10 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; m is either 0 or 1; Each n, q, and u is an integer between 0 and 16, independently of the others; Each p is an integer between 0 and 24, independently; Each s is an independent integer between 0 and 16; Each t is independently 1, 2, 3, 4, 5, or 6; Each R' is independently selected from H, C(O)OH, (CH2)OH, and NHAc; Each R'' is independently selected from H and CH3.

[0081] In one embodiment, each nitrogen atom and α-carbon atom on the peptide skeleton is optionally substituted with methyl atoms; Here, the variable group R 0 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , or R 10 However, when defined as a side chain of a cyclic amino acid, the corresponding amino acid nitrogen in the peptide backbone of formula I forms part of the cyclic group; Cyclic peptides may contain radionuclides as desired.

[0082] In one embodiment, the cyclic peptide is the cyclic peptide of formula I': [ka] or a pharmaceutically acceptable salt thereof.

[0083] In another embodiment, the cyclic peptide of formula B is provided herein: [ka] or a pharmaceutically acceptable salt thereof, P 1 is, -L 1 -Chelating agents, [ka] Selected from; D 1 It is a -NR”-chelating agent; L 1 It does not exist, or [ka] Selected from; Here, L 1 The amino group is P 1 Alternatively, it can bond to the carbonyl group of the chelating agent to form an amide bond; P 2 C(O)NH2, C(O)OH, [ka] Selected from; D 2 It is either OH or NH2; L 2 It does not exist, or [ka] Selected from; Here, L 2 The amino group is P 2 It links to the carbonyl group to form an amide bond; P 3 H, [ka] Selected from; L 3 It does not exist, or independently, [ka] Selected from; Here, L 3 The carbonyl group is P 2 It links to the amine group to form an amide bond; D 3 These are independently CH3, C(O)OH, and [ka] Selected from; X is a halogen, R 0 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 1 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 2 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 3 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; B 1 C 1~6 It is alkylene; C 1 C 1~6 It is alkylene; A 1 teeth, [ka] Selected from; where w is selected from 1, 2, or 3; R 4 This refers to the amino acid side chain of a natural amino acid or a non-natural amino acid, both of which are CH2C(O)OH or C(O)(CH2CH2O). p (CH2)2N(CH3)3 + It is arbitrarily replaced by; R 5 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 6 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 7 teeth, (i) amino acid side chains of natural amino acids, (ii) Selected from the amino acid side chains of non-natural amino acids, (iii) [ka] Selected from the group consisting of; R 8This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 9 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R 10 This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; m is either 0 or 1; Each n, q, and u is an integer between 0 and 16, independently of the others; Each p is an integer between 0 and 24, independently; Each s is an independent integer between 0 and 16; Each t is independently 1, 2, 3, 4, 5, or 6; Each R' is independently selected from H, C(O)OH, (CH2)OH, and NHAc; Each R'' is independently selected from H and CH3.

[0084] In one embodiment, each nitrogen atom and α-carbon atom on the peptide skeleton is optionally substituted with methyl atoms; Here, the variable group R 0 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , or R 10 However, when defined as a side chain of a cyclic amino acid, the corresponding amino acid nitrogen in the peptide backbone of formula B forms part of the cyclic group; Cyclic peptides may contain radionuclides as desired.

[0085] In one embodiment, the cyclic peptide of formula B is the cyclic peptide of B': [ka] or a pharmaceutically acceptable salt thereof, In the formula, each R 0a , R 1a , R 2a, R 3a , R 4a , R 5a , R 6a , R 7a , R 8a , R 9a , R 10a , R 1b or R 2b Each instance is independently selected from H and CH3. In one embodiment, R 0a , R 1a , R 2a , R 3a , R 4a , R 5a , R 6a , R 7a , R 8a , R 9a , R 10a , R 1b , or R 2b Of these, 0, 1, 2, 3, or 4 are CH3. In another embodiment of formula B', R 3a In yet another embodiment of formula B', R 5a This is CH3.

[0086] In one embodiment, the cyclic peptide of formula B is the cyclic peptide of formula Bi: [ka] or a pharmaceutically acceptable salt thereof, In the formula, R'' is independently selected from H and CH3. In one embodiment of the cyclic peptide of formula Bi or a pharmaceutically acceptable salt thereof, n is 3 and p is 0. In another embodiment of the cyclic peptide of formula Bi or a pharmaceutically acceptable salt thereof, L 1 It does not exist.

[0087] In another embodiment, the cyclic peptide of formula B is the cyclic peptide of formula Bii: [ka] or a pharmaceutically acceptable salt thereof, In the formula, R'' is independently selected from H and CH3. In one embodiment of the cyclic peptide of formula Bii or a pharmaceutically acceptable salt thereof, R' is H and n is 3. In another embodiment of the cyclic peptide of formula Bi or a pharmaceutically acceptable salt thereof, L 1 It does not exist.

[0088] In another embodiment, the cyclic peptide of formula B is the cyclic peptide of formula Biii: [ka] or a pharmaceutically acceptable salt thereof, In the formula, R'' is independently selected from H and CH3.

[0089] In one embodiment, the cyclic peptide of formula B is the cyclic peptide of formula Biv: [ka] or a pharmaceutically acceptable salt thereof, In the formula, each R is independently selected from H, CH3, OH, halogen, C(O)OH, and N(R)2 each time it appears.

[0090] In one embodiment, the cyclic peptide of formula B is the cyclic peptide of formula Ia: [ka] or a pharmaceutically acceptable salt thereof.

[0091] In another embodiment, the cyclic peptide of formula B is the cyclic peptide of formula Ib: [ka] or a pharmaceutically acceptable salt thereof.

[0092] In yet another embodiment, P 1 teeth, [ka] , -L 1 -Chelating agents, and [ka] Selected from; During the ceremony, D 1 It is an NR"-chelating agent; L 1 It does not exist or [ka] and; n and s are independent integers between 2 and 15; p is 8, 9, 10, 11, or 12.

[0093] In yet another embodiment, P 1 is, -L 1 -It is a chelating agent.

[0094] In one embodiment, P 1 is a chelating agent, [ka] Selected from.

[0095] In one embodiment, P 1 DOTA, [ka] Selected from.

[0096] In another embodiment, P 2 The compound is selected from C(O)NH2 and C(O)OH.

[0097] In yet another embodiment, P 2C(O)NH2, C(O)OH, [ka] Selected from; P 3 Ac and [ka] Selected from; X is a halo.

[0098] In another embodiment, the herein provides a cyclic peptide of formula B, where D 3 teeth, [ka] And, Here, w is selected from 1, 2, or 3; other variables are defined herein.

[0099] In one embodiment, P 3 is Ac or [ka] That is the case.

[0100] In yet another embodiment, the cyclic peptide of formula B is the cyclic peptide of formula II: [ka] or a pharmaceutically acceptable salt thereof, In the formula, P 2 This is either C(O)NH2 or C(O)OH.

[0101] In yet another embodiment, m is 0; P 1 DOTA, [ka] Selected from; P 2 C(O)NH2, C(O)OH, [ka] Selected from; P 3 is Ac or [ka] and; X is a halo; R 1 This is selected from the group consisting of amino acid side chains of Trp, 2Nal, 1Nal, 4CF3-Phe, 7Aza-Trp, 1Me-Trp, 5OH-Trp, BIP, 5Ome-Trp, 4F-Phe, 3Pya, 4Pya, PAF, MAF, OAF, 5Qui, 7MeO-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, D-Ala, Ala, α-Me-Trp, and NMe-Trp; R 2 It is selected from the group consisting of amino acid side chains of Thr, D-Ala, Ala, α-Me-Thr, Lys, and NMe-Thr; R 3 This is selected from the group consisting of amino acid side chains of Ile, Env, CHA, CBA, Nle, Tbg, THPG, Chg, 2Nal, 1Nal, 2CF3-Phe, 2PhEt-Ala, D-Ala, Ala, Leu, t-Bu-Ala, NMe-Nle, α-tert-amyl Gly, Allo-Ile, Lys(C12), Lys(C14), Lys(C16), α-Me-Ile, and NMe-tBuAla; A 1 teeth, [ka] Selected from the group consisting of; R 4 Asn, D-Ala, Ala, DAB-4-NHCOC5H11 DAB-4-NHCOC7H 15 Selected from the group consisting of amino acid side chains of Asp, Ser, Lys, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, 3Pya, 4Pya, Glu, NMe-Asn, Pip(CH2CO2H)Ala, and Pip(PegNMe3)Ala; R 5 It is selected from the group consisting of amino acid side chains of Asn, Ala, D-Ala, Trp, Asp, Lys, 3Pya, 4Pya, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, Glu, NMe-Asn, and Ser; R 6 , is selected from the group consisting of amino acid side chains of Trp, 4CF3-Phe, 1Me-Trp, 7Aza-Trp, BIP, 2Nal, 1Nal, α-Me-Trp, D-Ala, Ala, 4F-Phe, 5F-Trp, 5Ome-Trp, Asn, 5OH-Trp, 7Me-Trp, 7MeO-Trp, 7Cl-Trp, and NMe-Trp; R 7 teeth, (i) Selected from the group consisting of amino acid side chains of 3Pya, 4Pya, Lys(Me)3, His, Ala, D-Ala, Gln, Lys, Glu, Arg, Orn, NMe-His, and Ser; or (ii) [ka] Selected from the group consisting of; Each s is independently 3, 5, 10, 12, or 14; R 8 It is selected from the group consisting of amino acid side chains of Asp, D-Ala, Ala, Asn, Thr, NMe-Asp, and α-Me-Asp; R 9It is selected from the group consisting of amino acid side chains of Trp, 7aza-Trp, 1Me-Trp, D-Ala, Ala, 4F-Phe, 1Nal, 2Nal, 5F-Trp, 5Ome-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp; R 10 This is selected from the group consisting of amino acid side chains of Pro, D-Ala, Ala, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, Pip, 5,5-di-Me-Pro, NMe-Ser, trans4NH2-Pro, cis4NH2-Pro, Sar, Aze, NMe-Ala, NMe-Leu, R-3Me-Aze, α-Me-Aze, ACI, and 3Me2-Aze.

[0102] In another embodiment, m is 1; P 1 DOTA, [ka] Selected from; P 2 C(O)NH2, C(O)OH, [ka] Selected from; P 3 Ac and [ka] Selected from; X is a halo; R 0 It is selected from the group consisting of amino acid side chains of Gly, Met, D-Ala, Ala, Nle, and Nva; R 1This is selected from the group consisting of amino acid side chains of Trp, 2Nal, 1Nal, 4CF3-Phe, 7Aza-Trp, 1Me-Trp, 5OH-Trp, BIP, 5Ome-Trp, 4F-Phe, 3Pya, 4Pya, PAF, MAF, OAF, 5Qui, 7MeO-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, D-Ala, Ala, α-Me-Trp, and NMe-Trp; R 2 It is selected from the group consisting of amino acid side chains of Thr, D-Ala, Ala, α-Me-Thr, Lys, and NMe-Thr; R 3 This is selected from the group consisting of amino acid side chains of Ile, Env, CHA, CBA, Nle, Tbg, THPG, Chg, 2Nal, 1Nal, 2CF3-Phe, 2PhEt-Ala, D-Ala, Ala, Leu, t-Bu-Ala, NMe-Nle, α-tert-amyl Gly, Allo-Ile, Lys(C12), Lys(C14), Lys(C16), α-Me-Ile, and NMe-tBuAla; A 1 teeth, [ka] Selected from the group consisting of; R 4 Asn, D-Ala, Ala, DAB-4-NHCOC5H 11 DAB-4-NHCOC7H 15 Selected from the group consisting of amino acid side chains of Asp, Ser, Lys, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, 3Pya, 4Pya, Glu, and NMe-Asn; R 5 It is selected from the group consisting of amino acid side chains of Asn, Ala, D-Ala, Trp, Asp, Lys, 3Pya, 4Pya, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, Glu, NMe-Asn, and Ser; R 6This is selected from the group consisting of amino acid side chains of Trp, 4CF3-Phe, 1Me-Trp, 7Aza-Trp, BIP, 2Nal, 1Nal, α-Me-Trp, D-Ala, Ala, 4F-Phe, 5F-Trp, 5Ome-Trp, Asn, 5OH-Trp, 7Me-Trp, 7MeO-Trp, 7Cl-Trp, and NMe-Trp; R 7 is selected from the group consisting of amino acid side chains of 3Pya, 4Pya, Lys(Me)3, His, Ala, D-Ala, Gln, Lys, Glu, Arg, Orn, NMe-His, and Ser; or R 7 teeth, [ka] Selected from the group consisting of; Each s is independently 3, 5, 10, 12, or 14; R 8 It is selected from the group consisting of amino acid side chains of Asp, D-Ala, Ala, Asn, Thr, NMe-Asp, and α-Me-Asp; R 9 It is selected from the group consisting of amino acid side chains of Trp, 7aza-Trp, 1Me-Trp, D-Ala, Ala, 4F-Phe, 1Nal, 2Nal, 5F-Trp, 5Ome-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp; R 10 This is selected from the group consisting of amino acid side chains of Pro, D-Ala, Ala, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, Pip, 5,5-di-Me-Pro, NMe-Ser, trans4NH2-Pro, cis4NH2-Pro, Sar, Aze, NMe-Ala, NMe-Leu, R-3Me-Aze, α-Me-Aze, ACI, and 3Me2-Aze.

[0103] In one embodiment, m is 0 or 1; P 1is a chelating agent, [ka] Selected from the group consisting of; L 1 It does not exist, or [ka] Selected from the group consisting of; P 2 is either C(O)NH2 or C(O)OH; R 0 It is selected from the group consisting of amino acid side chains of Gly, Met, D-Ala, Ala, Nle, and Nva; R 1 This is selected from the group consisting of amino acid side chains of Trp, 2Nal, 1Nal, 4CF3-Phe, 7Aza-Trp, 1Me-Trp, 5OH-Trp, BIP, 5Ome-Trp, 4F-Phe, 3Pya, 4Pya, PAF, MAF, OAF, 5Qui, 7Ome-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, D-Ala, Ala, α-Me-Trp, and NMe-Trp; R 2 It is selected from the group consisting of amino acid side chains of Thr, D-Ala, Ala, α-Me-Thr, Lys, and NMe-Thr; R 3 This is selected from the group consisting of amino acid side chains of Ile, Env, CHA, CBA, Nle, Tbg, THPG, Chg, 2Nal, 1Nal, 2CF3-Phe, 2PhEt-Ala, D-Ala, Ala, Leu, t-Bu-Ala, NMe-Nle, α-tert-amyl Gly, Allo-Ile, Lys(C12), Lys(C14), Lys(C16), α-Me-Ile, and NMe-tBuAla; A 1 teeth, [ka] Selected from the group consisting of; R 4Asn, D-Ala, Ala, DAB-4-NHCOC5H 11 DAB-4-NHCOC7H 15 Selected from the group consisting of amino acid side chains of Asp, Ser, Lys(DOTA), Lys, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, 3Pya, 4Pya, Glu, NMe-Asn, Pip(CH2CO2H)Ala, Pip(PegNMe3)Ala, and Pip(GAE-DOTA)Ala; R 5 It is selected from the group consisting of amino acid side chains of Asn, Ala, D-Ala, Trp, Asp, Lys, Lys(DOTA), 3Pya, 4Pya, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, Glu, NMe-Asn, and Ser; R 6 This is selected from the group consisting of amino acid side chains of Trp, 4CF3-Phe, 1Me-Trp, 7Aza-Trp, BIP, 2Nal, 1Nal, aMe-Trp, D-Ala, Ala, 4F-Phe, 5F-Trp, 5Ome-Trp, Asn, 5OH-Trp, 7Me-Trp, 7MeO-Trp, 7Cl-Trp, and NMe-Trp; R 7 teeth, (i) Selected from the group consisting of amino acid side chains of 3Pya, 4Pya, Lys(Me)3, His, Ala, Phe, D-Ala, Gln, Lys, Glu, Arg, Orn, NMe-His, Trp, and Ser; or (ii) [ka] Selected from the group consisting of; s is 3, 5, 10, 12, or 14; R 8 It is selected from the group consisting of amino acid side chains of Asp, D-Ala, Ala, Asn, Thr, NMe-Asp, and α-Me-Asp; R 9It is selected from the group consisting of amino acid side chains of Trp, 7aza-Trp, 1Me-Trp, D-Ala, Ala, 4F-Phe, 1Nal, 2Nal, 5F-Trp, 5Ome-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp; R 10 This is selected from the group consisting of amino acid side chains of Pro, D-Ala, Ala, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, Pip, 5,5-di-Me-Pro, NMe-Ser, trans4NH2-Pro, cis4NH2-Pro, Sar, Aze, NMe-Ala, NMe-Leu, R-3Me-Aze, α-Me-Aze, ACI, and 3Me2-Aze.

[0104] In another embodiment, A 1 teeth, [ka] It is selected from the group consisting of the following.

[0105] In yet another embodiment, A 1 teeth, [ka] That is the case.

[0106] In yet another embodiment, P 1 DOTA, [ka] Selected from the group consisting of, Here, L 1 It does not exist, or [ka] It is selected from the group consisting of the following.

[0107] In yet another embodiment, m is 0.

[0108] In one embodiment, m is 0 or 1; P 1 teeth, [ka] Selected from the group consisting of; L 1 It does not exist, or [ka] Selected from the group consisting of; P 2 is either C(O)NH2 or C(O)OH; R 0 It is selected from the group consisting of amino acid side chains of Gly, Met, D-Ala, Ala, Nle, and Nva; R 1 This is selected from the group consisting of amino acid side chains of Trp, 2Nal, 1Nal, 7aza-Trp, 1Me-Trp, 5OH-Trp, BIP, 5Ome-Trp, 3Pya, 4Pya, 5Qui, 7Ome-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, α-Me-Trp, and NMe-Trp; R 2 It is selected from the group consisting of amino acid side chains of Thr, D-Ala, Ala, α-Me-Thr, Lys, and NMe-Thr; R 3 It is selected from the group consisting of amino acid side chains of Ile, Env, Nle, D-Ala, Ala, Leu, t-Bu-Ala, NMe-Nle, α-tert-amyl Gly, Allo-Ile, Lys(C12), Lys(C14), Lys(C16), α-Me-Ile, and NMe-tBuAla; A 1 teeth, [ka] Selected from the group consisting of; R 4 Asn, D-Ala, Ala, DAB-4-NHCOC5H 11 DAB-4-NHCOC7H 15 Selected from the group consisting of amino acid side chains of Asp, Ser, Lys(DOTA), Lys, 3-(4-piperidinyl)-Ala, PipA(acetic acid), 3-(1-morpholinyl)-Ala, 3Pya, Glu, and NMe-Asn; R 5 It is selected from the group consisting of amino acid side chains of Asn, Ala, D-Ala, Asp, Lys, Glu, NMe-Asn, and Ser; R 6 This is selected from the group consisting of amino acid side chains of Trp, 4CF3-Phe, 1Me-Trp, 7aza-Trp, BIP, 2Nal, 1Nal, aMe-Trp, 4F-Phe, 5F-Trp, 5Ome-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, 7Cl-Trp, and NMe-Trp; R 7 It is selected from the group consisting of amino acid side chains of 3Pya, 4Pya, Lys(Me)3, His, Ala, Phe, D-Ala, Gln, Lys, Glu, Arg, Orn, NMe-His, Trp, and Ser; R 8 It is selected from the group consisting of amino acid side chains of Asp, D-Ala, Ala, Asn, Thr, NMe-Asp, and α-Me-Asp; R 9 This is selected from the group consisting of amino acid side chains of Trp, 7aza-Trp, 1Me-Trp, 1Nal, 2Nal, 5F-Trp, 5Ome-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp; R 10 This is selected from the group consisting of amino acid side chains of Pro, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, 5,5-di-Me-Pro, trans4NH2-Pro, cis4NH2-Pro, Aze, ACI, and 3Me2-Aze.

[0109] In one embodiment, m is 0 or 1; P 1 teeth, [ka] Selected from the group consisting of; L 1 It does not exist, or [ka] Selected from the group consisting of; P 2 is either C(O)NH2 or C(O)OH; R 0 It is selected from the group consisting of amino acid side chains of Gly, Met, D-Ala, Ala, Nle, and Nva; R 1 This is selected from the group consisting of amino acid side chains of Trp, 7aza-Trp, 1Me-Trp, 5OH-Trp, 5Ome-Trp, 7Ome-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, α-Me-Trp, and NMe-Trp; R 2 It is selected from the group consisting of amino acid side chains of Thr, α-Me-Thr, Lys, and NMe-Thr; R 3 It is selected from the group consisting of amino acid side chains of D-Ala, Ala, t-Bu-Ala, and NMe-tBuAla; A 1 teeth, [ka] Selected from the group consisting of; R 4 It is selected from the group consisting of amino acid side chains of 3-(4-piperidinyl)-Ala, PipA(acetic acid), and 3-(1-morpholinyl)-Ala; R 5 It is selected from the group consisting of amino acid side chains of Asn, Asp, and NMe-Asn; R 6The amino acid side chains are selected from 2Na and 1Na; R 7 It is selected from the group consisting of amino acid side chains of 3Pya, 4Pya, Lys(Me)3, His, Ala, Phe, D-Ala, Gln, Lys, Glu, Arg, Orn, NMe-His, Trp, and Ser; R 8 It is selected from the group consisting of amino acid side chains of Asp, Asn, NMe-Asp, and α-Me-Asp; R 9 This is selected from the group consisting of amino acid side chains of Trp, 7aza-Trp, 1Me-Trp, 5F-Trp, 5Ome-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp; R 10 This is selected from the group consisting of amino acid side chains of Pro, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, 5,5-di-Me-Pro, trans4NH2-Pro, and cis4NH2-Pro.

[0110] In another embodiment, R 1 is the amino acid side chain of Trp; R 2 is the amino acid side chain of Thr; R 3 These are amino acid side chains of Ile or NMe-tBuAla; R 4 These are selected from the amino acid side chains of Lys, 3-(4-piperidinyl)-Ala, and PipA(acetic acid); R 5 This is an amino acid side chain of Asn or NMe-Asn; R 6 This is the amino acid side chain of Trp or 2NaI; R 7 These are amino acid side chains of His or Lys; R 8 It is the amino acid side chain of Asp; R 9 is the amino acid side chain of Trp; R 10 This is the amino acid side chain of Pro.

[0111] In another embodiment, R 1 is the amino acid side chain of Trp; R 2 is the amino acid side chain of Thr; R 3 is the amino acid side chain of Ile; R 4 These are selected from the amino acid side chains of Lys, 3-(4-piperidinyl)-Ala, and PipA(acetic acid); R 5 This is an amino acid side chain of Asn or NMe-Asn; R 6 It is a 2Na or Trp amino acid side chain; R 7 It is selected from the amino acid side chain of His; R 8 It is the amino acid side chain of Asp; R 9 is the amino acid side chain of Trp; R 10 This is the amino acid side chain of Pro.

[0112] In another embodiment, R 1 is the amino acid side chain of Trp; R 2 is the amino acid side chain of Thr; R 3 This is the amino acid side chain of NMe-tBuAla; R 4 It is the amino acid side chain of 3-(4-piperidinyl)-Ala or PipA(acetic acid); R 5 It is the amino acid side chain of Asn; R 6 It is the amino acid side chain of 2NaI; R 7 is the amino acid side chain of Lys; R 8 It is the amino acid side chain of Asp; R 9 is the amino acid side chain of Trp; R 10 This is the amino acid side chain of Pro.

[0113] In one embodiment, P 1 teeth, [ka] Selected from the group consisting of; L 1 It does not exist, or [ka] Selected from the group consisting of; P 2 is either C(O)NH2 or C(O)OH; R 1 This is selected from the group consisting of amino acid side chains of Trp, 7aza-Trp, 1Me-Trp, 5OH-Trp, 5Ome-Trp, 7Ome-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, α-Me-Trp, and NMe-Trp; R 2 is the amino acid side chain of Thr; R 3 It is selected from the group consisting of amino acid side chains of D-Ala, Ala, t-Bu-Ala, and NMe-tBuAla; A 1 teeth, [ka] and; B 1 It is CH2; C 1 It is CH2; R 4 It is selected from the group consisting of amino acid side chains of 3-(4-piperidinyl)-Ala, PipA(acetic acid), and 3-(1-morpholinyl)-Ala; R 5It is selected from the group consisting of amino acid side chains of Asn, Asp, and NMe-Asn; R 6 These are 2Na or 1Na amino acid side chains; R 7 It is selected from the group consisting of amino acid side chains of His, Lys, and Trp; R 8 It is selected from the group consisting of amino acid side chains of Asp, NMe-Asp, and α-Me-Asp; R 9 This is selected from the group consisting of amino acid side chains of Trp, 7aza-Trp, 1Me-Trp, 5F-Trp, 5Ome-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp; R 10 This is selected from the group consisting of amino acid side chains of Pro, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, 5,5-di-Me-Pro, trans4NH2-Pro, and cis4NH2-Pro.

[0114] In one embodiment, P 1 teeth, [ka] Selected from the group consisting of; L 1 It does not exist, or [ka] Selected from the group consisting of; P 2 is either C(O)NH2 or C(O)OH; R 1 This is selected from the group consisting of amino acid side chains of Trp, 7aza-Trp, 1Me-Trp, 5OH-Trp, 5Ome-Trp, 7Ome-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, α-Me-Trp, and NMe-Trp; R2 It is selected from the group consisting of amino acid side chains of Thr, α-Me-Thr, and NMe-Thr; R 3 It is selected from the group consisting of amino acid side chains of D-Ala, Ala, t-Bu-Ala, and NMe-tBuAla; A 1 teeth, [ka] and; B 1 It is CH2; C 1 It is CH2; R 4 It is selected from the group consisting of amino acid side chains of 3-(4-piperidinyl)-Ala, PipA(acetic acid), and 3-(1-morpholinyl)-Ala; R 5 This is an amino acid side chain of Asn or NMe-Asn; R 6 These are 2Na or 1Na amino acid side chains; R 7 It is selected from the group consisting of amino acid side chains of His, Lys, and Trp; R 8 It is selected from the group consisting of amino acid side chains of Asp, NMe-Asp, and α-Me-Asp; R 9 This is selected from the group consisting of amino acid side chains of Trp, 7aza-Trp, 1Me-Trp, 5F-Trp, 5Ome-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp; R 10 This is selected from the group consisting of amino acid side chains of Pro, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, 5,5-di-Me-Pro, trans4NH2-Pro, and cis4NH2-Pro.

[0115] In another embodiment, P 1 teeth, [ka] Selected from the group consisting of; L 1 It does not exist, or [ka] Selected from the group consisting of; P 2 It is C(O)NH2; R 1 is the amino acid side chain of Trp; R 2 is the amino acid side chain of Thr; R 3 These are amino acid side chains of Ile or NMe-tBuAla; A 1 teeth, [ka] and; B 1 It is CH2; C 1 It is CH2; R 4 is the amino acid side chain of Lys, 3-(4-piperidinyl)-Ala, or PipA (acetic acid); R 5 This is an amino acid side chain of Asn or NMe-Asn; R 6 This is the amino acid side chain of Trp or 2NaI; R 7 These are amino acid side chains of His or Lys; R 8 It is the amino acid side chain of Asp; R 9 is the amino acid side chain of Trp; R 10 This is the amino acid side chain of Pro.

[0116] In one embodiment, P 1 teeth, [ka] Selected from the group consisting of; L 1 It does not exist, or [ka] Selected from the group consisting of; P 2 It is C(O)NH2; R 1 is the amino acid side chain of Trp; R 2 is the amino acid side chain of Thr; R 3 is the amino acid side chain of Ile; A 1 teeth, [ka] and; B 1 It is CH2; C 1 It is CH2; R 4 It is the amino acid side chain of 3-(4-piperidinyl)-Ala or PipA(acetic acid); R 5 This is an amino acid side chain of Asn or NMe-Asn; R 6 It is a 2Na amino acid side chain; R 7 His is the amino acid side chain; R 8 It is the amino acid side chain of Asp; R 9 is the amino acid side chain of Trp; R 10 This is the amino acid side chain of Pro.

[0117] In one embodiment, P 1 teeth, [ka] Selected from the group consisting of; L 1 It does not exist, or [ka] Selected from the group consisting of; P 2 It is C(O)NH2; R 1 is the amino acid side chain of Trp; R 2 is the amino acid side chain of Thr; R 3 This is the amino acid side chain of NMe-tBuAla; A 1 teeth, [ka] and; B 1 It is CH2; C 1 It is CH2; R 4 It is the amino acid side chain of 3-(4-piperidinyl)-Ala or PipA(acetic acid); R 5 It is the amino acid side chain of Asn; R 6 It is a 2Na amino acid side chain; R 7 is the amino acid side chain of Lys; R 8 It is the amino acid side chain of Asp; R 9 is the amino acid side chain of Trp; R 10 This is the amino acid side chain of Pro.

[0118] In another embodiment, R 0This is selected from the group consisting of amino acid side chains of Gly, Met, D-Ala, Ala, Nle, and Nva.

[0119] In yet another embodiment, R 1 This is selected from the group consisting of amino acid side chains of Trp, 2Nal, 1Nal, 4CF3-Phe, 7aza-Trp, 1Me-Trp, 5OH-Trp, BIP, 5Ome-Trp, 4F-Phe, 3Pya, 4Pya, PAF, MAF, OAF, 5Qui, 7MeO-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, D-Ala, Ala, α-Me-Trp, and NMe-Trp. In yet another embodiment, R 1 The amino acid side chains are selected from the group consisting of Trp, 7aza-Trp, 1Me-Trp, 5OH-Trp, 5Ome-Trp, 7Ome-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, α-Me-Trp, and NMe-Trp. In one embodiment, R 1 is the amino acid side chain of Trp or 7Me-Trp. In another embodiment, R 1 This is the amino acid side chain of Trp.

[0120] In yet another embodiment, R 2 R is selected from the group consisting of amino acid side chains of Thr, D-Ala, Ala, α-Me-Thr, Lys, and NMe-Thr. In yet another embodiment, R 2 R is selected from the group consisting of amino acid side chains of Thr, α-Me-Thr, and NMe-Thr. In one embodiment, R 2 This is the amino acid side chain of Thr.

[0121] In one embodiment, R 3 This is selected from the group consisting of amino acid side chains of Ile, Env, CHA, CBA, Nle, Tbg, THPG, Chg, 2Nal, 1Nal, 2CF3-Phe, 2PhEt-Ala, D-Ala, Ala, Leu, t-Bu-Ala, NMe-Nle, α-tert-amyl Gly, Allo-Ile, Lys(C12), Lys(C14), Lys(C16), α-Me-Ile, and NMe-tBuAla. In another embodiment, R3 R is selected from the group consisting of amino acid side chains of Ile, Allo-Ile, α-Me-Ile, and NMe-tBuAla. In yet another embodiment, R 3 R is selected from the group consisting of amino acid side chains of D-Ala, Ala, t-Bu-Ala, and NMe-tBuAla. In yet another embodiment, R 3 is an amino acid side chain of Ile or NMe-tBuAla. In one embodiment, R 3 is the amino acid side chain of Ile. In another embodiment, R 3 This is the amino acid side chain of NMe-tBuAla.

[0122] In yet another embodiment, R 4 Asn, D-Ala, Ala, DAB-4-NHCOC5H 11 DAB-4-NHCOC7H 15 Selected from the group consisting of the amino acid side chains of Asp, Ser, Lys, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, 3Pya, 4Pya, Glu, NMe-Asn, Pip(CH2CO2H)Ala, and Pip(PegNMe3)Ala. In yet another embodiment, R 4 This is selected from the group consisting of the amino acid side chains of 3-(4-piperidinyl)-Ala, PipA (acetic acid), and 3-(1-morpholinyl)-Ala. In another embodiment, R 4 This is the amino acid side chain of 3-(4-piperidinyl)-Ala or PipA (acetic acid).

[0123] In yet another embodiment, R 5 This is selected from the group consisting of amino acid side chains of Asn, Ala, D-Ala, Trp, Asp, Lys, 3Pya, 4Pya, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, Glu, NMe-Asn, and Ser. In yet another embodiment, R 5 is an amino acid side chain of Asn or NMe-Asn. In one embodiment, R 5 R is the amino acid side chain of NMe-Asn. In another embodiment, R 5 This is the amino acid side chain of Asn.

[0124] In yet another embodiment, R 6 This is selected from the group consisting of amino acid side chains of Trp, 4CF3-Phe, 1Me-Trp, 7Aza-Trp, BIP, 2Nal, 1Nal, α-Me-Trp, D-Ala, Ala, 4F-Phe, 5F-Trp, 5MeO-Trp, Asn, 5OH-Trp, 7Me-Trp, 7MeO-Trp, 7Cl-Trp, and NMe-Trp. In yet another embodiment, R 6 The amino acid side chains are selected from the group consisting of Trp, 1Me-Trp, 7aza-Trp, 2Nal, 1Nal, α-Me-Trp, 5F-Trp, 5MeO-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, 7Cl-Trp, and NMe-Trp. In one embodiment, R 6 This is selected from the group consisting of Trp, 2Na, and 1Na amino acid side chains. In another embodiment, R 6 is the amino acid side chain of Trp. In yet another embodiment, R 6 This is the amino acid side chain of 2NaI.

[0125] In yet another embodiment, R 7 The amino acid side chains are selected from the group consisting of 3Pya, 4Pya, Lys(Me)3, His, Ala, D-Ala, Gln, Lys, Glu, Arg, Orn, NMe-His, and Ser. In one embodiment, R 7 is an amino acid side chain of His or Lys. In one embodiment, R 7 This is the amino acid side chain of His. In one embodiment, R 7 This is the amino acid side chain of Lys.

[0126] In yet another embodiment, R 8 This is selected from the group consisting of amino acid side chains of Asp, D-Ala, Ala, Asn, Thr, NMe-Asp, and α-Me-Asp. In one embodiment, R 8 This is selected from the group consisting of amino acid side chains of Asp, Asn, NMe-Asp, and α-Me-Asp. In another embodiment, R 8This is selected from the group consisting of amino acid side chains of Asp, NMe-Asp, and α-Me-Asp. In yet another embodiment, R 8 This is the amino acid side chain of Asp.

[0127] In one embodiment, R 9 The amino acid side chains are selected from the group consisting of Trp, 7aza-Trp, 1Me-Trp, D-Ala, Ala, 4F-Phe, 1Nal, 2Nal, 5F-Trp, 5MeO-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp. In one embodiment, R 9 The amino acid side chains are selected from the group consisting of Trp, 7aza-Trp, 1Me-Trp, 5F-Trp, 5Ome-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp. In one embodiment, R 9 This is the amino acid side chain of Trp.

[0128] In one embodiment, R 10 The amino acid side chains are selected from the group consisting of Pro, D-Ala, Ala, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, Pip, 5,5-di-Me-Pro, NMe-Ser, trans4NH2-Pro, cis4NH2-Pro, Sar, Aze, NMe-Ala, NMe-Leu, R-3Me-Aze, α-Me-Aze, ACI, and 3Me2-Aze. In one embodiment, R 10 The amino acid side chains are selected from the group consisting of Pro, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, 5,5-di-Me-Pro, trans4NH2-Pro, and cis4NH2-Pro. In one embodiment, R 10 This is the amino acid side chain of Pro.

[0129] In another embodiment, P 1 teeth, [ka] Selected from the group consisting of; L 1 It does not exist, or [ka] It is selected from the group consisting of the following.

[0130] In another embodiment, P 1 teeth, [ka] That is the case.

[0131] In yet another embodiment, B 1 is CH2 or C(CH3)2; C 1 It is either CH2 or C(CH3)2.

[0132] In yet another embodiment, B 1 It is CH2; C 1 This is CH2.

[0133] In yet another embodiment, the chelating agent is selected from the chelating agents in Table C.

[0134] In one embodiment, the chelating agent is independently ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 1,4,7,10-tetraazacyclododecane-N,N',N”,N”'-tetraacetic acid (DOTA), 6-((16-((6-carboxypyridine-2-yl)methyl)-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane-7-yl)methyl)-4-isothiocyanatepico Phosphate (Macropa), Macrodipa, 2,2',2",2"'-(1,10-dioxa-4,7,13,16-tetraazacyclooctadecane-4,7,13,16-tetrayl)tetraacetic acid) (Crown), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, α-(2-carboxyethyl) (DOTAGA), 1,4,7-triazacyclo Selected from the group consisting of nonane-N,N',N''-triacetic acid (NOTA), 1,4,7,10-tetraazacyclododecane-N,N',N”,N'''-tetraacetic acid (TETA), 1,4,7,10,13-pentazacyclopentadecane-N,N',N”,N''',N'''-pentaacetic acid (PEPA), and 1,4,7,10,13,16-hexaazacyclohexadecane-N,N',N”,N''',N””,N'''-hexaacetic acid (HEHA). In another embodiment, the chelating agent is selected from deferoxamine (DFO), 5,11,16,22-tetraazahexacosanediamide (DFO*), and N,N'-1,4-butanediylbis[N-[3-[[(1,6-dihydro-1-hydroxy-6-oxo-2-pyridinyl)carbonyl]amino]propyl]-1,6-dihydro-1-hydroxy-6-oxo-2-pyridinecarboxamide](HOPO).

[0135] In another embodiment, the chelating agent is DOTA. In yet another embodiment, the chelating agent is DOTAGA. In yet another embodiment, the chelating agent is Macrodipa. In one embodiment, the chelating agent is Macropa.

[0136] In another embodiment, formula B is substituted with at least one chelating agent. In yet another embodiment, formula B is substituted with one chelating agent. In yet another embodiment, formula B is substituted with two chelating agents.

[0137] In the formulas provided herein, the variable (e.g., R) 0 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , or R 10 The group can be defined as a cyclic amino acid, for example, a side chain of proline. In that case, the corresponding amino acid nitrogen of the peptide skeleton of the general formula provided herein forms part of the cyclic group. For example, "R 10 "Selected from the group consisting of amino acid side chains such as Pro, α-Me-Pro, trans4-fluoro-Pro, and cis4-fluoro-Pro" is defined as follows: [ka] In this representation, each R is independently either a hydrogen atom or a substituent.

[0138] In another embodiment, the compound of formula B is selected from the group consisting of compounds from Table A.

[0139] [Table 2]

[0140] [Table 3]

[0141] [Table 4]

[0142] Table 5

[0143] Table 6

[0144] Table 7

[0145] Table 8

[0146] Table 9

[0147] Table 10

[0148] Table 11

[0149] Table 12

[0150] Table 13

[0151] Table 14

[0152] Table 15

[0153] [Table 16]

[0154] [Table 17]

[0155] In yet another embodiment, the cyclic peptide of formula B is selected from the peptides in Table B.

[0156] [Table 18]

[0157] In another embodiment, the herein provides a pharmaceutical composition comprising a peptide described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0158] The compounds disclosed herein may exist as tautomers and optical isomers (e.g., enantiomers, diastereomers, diastereomer mixtures, racemic or non-racemic mixtures, etc.). Absolute stereochemistry is specified according to the Kahn-Ingold-Prelogue RS system. Chiral centers with known absolute configurations are denoted by the prefixes R and S, assigned by standard sequencing rules and preceded by appropriate locant as needed (Pure & Appl. Chem. 45, 1976, 11-30). Certain examples include chemical structures indicated or represented as (R*) or (S*). When (R*) or (S*) is used in the name or chemical expression of a compound, it is intended to convey that the compound is a pure monoisomer at its stereocenter, but the absolute configuration of that stereocenter has not been established. Therefore, a compound designated as (R*) refers to a compound that is a pure single isomer at its stereocenter having the absolute configuration of I or (S), and a compound designated as (S*) refers to a compound that is a pure single isomer at its stereocenter having the absolute configuration of (R) or (S).

[0159] The compounds provided herein may also contain all isotopes of atoms present in the intermediate or final compound. Isotopes include atoms with the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds of the present invention may be replaced or substituted with isotopes of atoms in natural or unnatural abundance. In some embodiments, the compound contains at least one deuterium atom. For example, one or more hydrogen atoms in the compounds of this disclosure may be replaced or substituted with deuterium. In some embodiments, the compound contains two or more deuterium atoms. In some embodiments, the compound contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Methods for synthesizing isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, NY, Appleton-Century-Crofts, 1971; The Renaissance of H / D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling by James R. Hanson, Royal Society of Chemistry, 2011). Isotopic-labeled compounds can be used in various tests such as NMR spectroscopy, metabolic experiments, and / or assays.

[0160] In the compounds provided herein, any atom not specifically designated as a particular isotope represents any stable isotope of that atom. Unless otherwise specified, where a position is specifically designated as "H" or "hydrogen," that position is understood to have hydrogen in its isotopic composition at its natural abundance. Similarly, where a position is specifically designated as "D" or "deuterium," that position is understood to have deuterium at an abundance at least 3000 times its natural abundance of 0.015% (i.e., at least 45% of deuterium incorporated).

[0161] In embodiments, the compounds provided herein have an isotopic enrichment factor of at least 3500 (52.5% deuterium incorporation into each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation) for each designated deuterium atom.

[0162] Examples of crosslinking moieties / peptide staples for use with the compounds of this disclosure include, but are not limited to, amide-based (e.g., lactam) crosslinks; aromatic ring-based crosslinks; hydrocarbon chains; alkene-based hydrocarbon crosslinks (e.g., using Fmoc-'-2-(2'-pentenyl)alanine); triazole-based click crosslinks, e.g., copper(I)-catalyzed hysgene 1,3-dipolar cycloaddition reaction between side-chain azide and alkynyl moieties (e.g., Fmoc-L-Nle(εN3) and Fmoc-D-Pra) (see S. Kawamoto, et al., J. Med. Chem. 2012, 55(3), 1137-1146); dialkynyl staples for stapling linear diazide-peptides (e.g., 1,4-diethynylbenzene, diethynylpentane, diethynylamine); sulfide-bonded disulfide, thioether and bis-thioether crosslinks; perfluorobenzene crosslinks; or combinations thereof.

[0163] In some embodiments, the crosslinked portion comprises an amide bond between an amine functional group and a carboxylate functional group, which are present in the side chains of an amino acid, a non-natural amino acid, or a non-amino acid residue, respectively. In some embodiments, the amine or carboxylate functional group is part of a non-amino acid residue or a non-natural amino acid residue. In some embodiments, the crosslinked portion comprises an amide bond produced by the reaction of the side chains of the following amino acid pairs: lysine and glutamate; lysine and aspartate; ornithine and glutamate; ornithine and aspartate; homolysine and glutamic acid; homolysine and aspartic acid; and other combinations of amino acids, non-natural amino acids, or non-amino acid residues, including primary amines and carboxylic acids. In some embodiments, the crosslinked portion is formed by a cyclization reaction using olefin metathesis.

[0164] In some embodiments, the crosslinking moiety includes a disulfide bond formed between two thiol-containing residues. In some embodiments, the crosslinking moiety includes one or more thioether bonds. Such thioether bonds may include those found in cyclo-thioalkyl compounds. These bonds may be formed during a chemical cyclization reaction between a chloroacetic acid N-terminal modification group and a cysteine ​​residue. In some embodiments, the crosslinking moiety includes one or more triazole rings.

[0165] In some embodiments, the crosslinked portion comprises one or more hydrocarbon chains (linear or branched) and / or hydrocarbon rings (cyclic, heterocyclic, aromatic, or heteroaromatic). In some embodiments, the hydrocarbon crosslinked portion may be introduced by reaction with a reagent containing a plurality of reactive halides, including, but not limited to, poly(bromomethyl)benzene, poly(bromomethyl)pyridine, poly(bromomethyl)alkylbenzene, and / or (E)-1,4-dibromobuta-2-ene. Examples of poly(bromomethyl)benzene molecules of this disclosure include 1,2-bis(bromomethyl)benzene; 1,3-bis(bromomethyl)benzene; and 1,4-bis(bromomethyl)benzene.

[0166] In some embodiments, the thiol group of a cysteine ​​residue is crosslinked with another cysteine ​​residue to form a disulfide bond. In some embodiments, the thiol group of a cysteine ​​residue reacts with the bromomethyl group of a poly(bromomethyl)benzene molecule to form a stable bond (see, for example, Timmerman et al., ChemBioChem (2005) 6:821-824, the contents of which are incorporated herein by reference in their entirety).

[0167] In some embodiments, bis-, tris-, and tetrakis(bromomethyl)benzene molecules can be used to generate crosslinking moieties, producing peptides having one, two, or three loops, respectively. The bromomethyl groups of the poly(bromomethyl)benzene molecules may be positioned on the benzene ring on adjacent ring carbons (ortho- or o-), with the ring carbon separating two groups (meta- or m-), or on opposite ring carbons (para- or p-). In some embodiments, m-bis(bromomethyl)benzene (i.e., m-dibromoxylen), o-bis(bromomethyl)benzene (i.e., o-dibromoxylen), and / or p-bis(bromomethyl)benzene (i.e., p-dibromoxylen) are used to form cyclic peptides. In some embodiments, the thiol groups of the cysteine ​​residues react with other reagents containing one or more bromo-functional groups to form stable bonds. Examples of such reagents, though not limited to them, include poly(bromomethyl)pyridine (e.g., 2,6-bis(bromomethyl)pyridine), poly(bromomethyl)alkylbenzene (e.g., 1,2-bis(bromomethyl)-4-alkylbenzene), and / or (E)-1,4-dibromobuta-2-ene.

[0168] In some embodiments, the side-chain amino groups and terminal amino groups are crosslinked with disuccinimidyl glutarate (see, e.g., Millward et al., J.Am.Chem.Soc. (2005) 127:14142-14143). In some embodiments, thioether bonds are generated using an enzymatic method that depends on the reaction between (1) cysteine ​​and (2) a dehydroalanine or dehydrobutyrine group catalyzed by lanthibiotic synthetase (see, e.g., Levengood et al., Bioorg. and Med.Chem.Lett. (2008) 18:3025-3028). Dehydro functional groups can also be chemically generated by oxidation of selenium-containing amino acid side chains incorporated during translation (see, e.g., Seebeck et al., J.Am.Chem.Soc. 2006).

[0169] In some embodiments, the crosslinking portion includes an aromatic six-membered ring (e.g., benzene). In some embodiments, the crosslinking portion includes a heterocyclic six-membered ring containing one nitrogen atom (e.g., pyridine). In some embodiments, the crosslinking portion includes a heterocyclic six-membered ring containing two nitrogen atoms (e.g., pyridazine, pyrimidine, pyrazine). In some embodiments, the crosslinking portion includes a heterocyclic six-membered ring containing three nitrogen atoms (e.g., triazane). In some embodiments, the crosslinking portion includes a heterocyclic five-membered ring containing one nitrogen atom (e.g., pyrrole). In some embodiments, the crosslinking portion includes a heterocyclic five-membered ring containing two nitrogen atoms (e.g., imidazole, pyrazole). In some embodiments, the crosslinking portion includes a heterocyclic five-membered ring containing three nitrogen atoms (e.g., triazole).

[0170] The peptides of this disclosure may be cyclized via a carboxyl terminus, an amino terminus, or any other convenient binding site, such as the sulfur of cysteine ​​(e.g., via the formation of a disulfide bond between two cysteine ​​residues in the sequence) or any side chain of an amino acid residue. Further bonds that form a cyclic loop include maleimide bonds, amide bonds, ester bonds, ether bonds, thiol ether bonds, hydrazone bonds, or acetamide bonds.

[0171] In some embodiments, the peptides of this disclosure are formed using a lactam moiety. Such cyclic peptides may be formed, for example, by synthesis on a solid support Wang resin using standard Fmoc chemistry. In some cases, Fmoc-Asp(allyl)-OH and Fmoc-Lys(alloc)-OH are incorporated into the peptide to function as precursor monomers for lactam crosslinking.

[0172] In some embodiments, the peptides of the disclosure are linear peptides. In some embodiments, the peptides of the disclosure are cyclic peptides. In some embodiments, the cyclic peptides include disulfide bonds. In some embodiments, the peptides of the disclosure are linear peptides before the cyclization step. In some embodiments, the peptides of the disclosure are linear peptides before the formation of disulfide bonds.

[0173] Generally, disulfide bond formation involves a reaction between the sulfhydryl (SH) side chains of two cysteine ​​residues. Proper disulfide bonds provide stability to proteins and reduce further entropy selection that facilitates folding to the native state by restricting unfolded or improperly folded conformations.

[0174] Terminal modification and conjugation One method for protecting peptides from proteolysis involves chemical modification or "capping" of the amino-terminus and / or carboxy-terminus of the peptide. As used herein, the terms “chemically modified” or “capped” are used synonymously to refer to the introduction of blocking groups at one or both ends of a compound by covalent modification. Suitable blocking groups serve to block the ends of the peptide without reducing the peptide’s biological activity. Any residue located at the amino-terminus, carboxy-terminus, or both ends of the compounds described may be chemically modified. In some embodiments, the peptides of this disclosure include N-terminal and / or C-terminal modifications.

[0175] In one embodiment, the amino terminus of a compound is chemically modified by acetylation to produce an N-acetylated peptide (which may be represented by "Ac-" in the structure or formula of this disclosure). In another embodiment, the carboxyl terminus of the peptide described is chemically modified by amidation to obtain a primary carboxamide at the C terminus (which may be represented by "amide" in the peptide sequence, structure or claims of this disclosure). In some embodiments, both the amino and carboxyl terms are chemically modified by acetylation and amidation, respectively. However, other capping groups are also possible. For example, the amino terminus may be capped by acylation with a group such as an acetyl group or a benzoyl group, or with a natural or unnatural amino acid such as β-alanine, or by alkylation with an acetyl group, or with a group such as a benzyl or butyl group, or by sulfonylation to produce a sulfonamide. Similarly, the carboxyl terminus may be esterified or converted to secondary amides and acylsulfonamides, etc.

[0176] In some embodiments, the N-terminal capping functional group is bonded to a terminal amino group and can be selected from the following group: formyl; alkanoyl having 1 to 10 carbon atoms, e.g., acetyl, propionyl, butyryl; alkenoyl having 1 to 10 carbon atoms, e.g., hexa-3-enoyl; alkinoyl having 1 to 10 carbon atoms, e.g., hexa-5-inoyl; aroyl, e.g., benzoyl or 1-naphthoyl; heteroaloyl, e.g., 3-pyroyl or 4-quinoyl; alkylsulfonyl, e.g., methanesulfonyl; Aryl sulfonyls, e.g., benzenesulfonyl or sulfanillyl; heteroarylsulfonyls, e.g., pyridine-4-sulfonyl; substituted alkanoyls having 1 to 10 carbon atoms, e.g., 4-aminobutyryl; substituted alkenoyls having 1 to 10 carbon atoms, e.g., 6-hydroxy-hexa-3-enoyl; substituted alquinoyls having 1 to 10 carbon atoms, e.g., 3-hydroxy-hexa-5-inoyl; substituted alloyls, e.g., 4-chlorobenzoyl or 8-hydroxy-naphtha-2-oil; substituted heteroaloyls, e.g., 2,4-Dioxo-1,2,3,4-Tetrahydro-3-methylquinazoline-6-oil; Substituted alkylsulfonyl, e.g., 2-aminoethanesulfonyl; Substituted arylsulfonyl, e.g., 5-dimethylamino-1-naphthalenesulfonyl; Substituted heteroarylsulfonyl, e.g., 1-methoxy-6-isoquinolinesulfonyl; Carbamoyl or thiocarbamoyl; Substituted carbamoyl (R'-NH-CO) or Substituted thiocarbamoyl (R'-NH-CS) (where R' is alkyl, alkenyl, alkynyl, aryl, heteroaryl, Substituting alkyl, substituted alkenyl, substituted alkynyl, substituted aryl, or substituted heteroaryl); substituted carbamoyl (R'-NH-CO) and substituted thiocarbamoyl (R'-NH-CS) (where R' is alkanoyl, alkenoyl, alkinoyl, aroyl, heteroaloyl, substituted alkanoyl, substituted alkenoyl, substituted alkinoyl, substituted aroyl, or substituted heteroaloyl, all as defined above); Lys-(Gly)n (where n=1 to 8); or Tyr-(Gly)n (where n=1 to 8).

[0177] In some embodiments, the C-terminal capping functional group may be either an amide bond with the terminal carboxyl or an ester bond with the terminal carboxyl.The capping functional group providing the amide bond is denoted as NR1R2, where R1 and R2 can be independently selected from the following group: hydrogen; alkyl groups having 1 to 10 carbon atoms, e.g., methyl, ethyl, isopropyl; preferably alkenyl groups having 1 to 10 carbon atoms, e.g., propa-2-enyl; preferably alkynyl groups having 1 to 10 carbon atoms, e.g., propa-2-inyl; substituted alkyl groups having 1 to 10 carbon atoms, e.g., hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, alpha Alkylthioalkynyl, halogenoalkyl, cyanoalkyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, alkanoylalkyl, carboxyalkyl, carbamoylalkyl; Substitutive alkenyls having 1 to 10 carbon atoms, e.g., hydroxyalkenyl, alkoxyalkenyl, mercaptoalkenyl, alkylthioalkenyl, halogenoalkenyl, cyanoalkenyl, aminoalkenyl, alkylaminoalkenyl, dialkylaminoalkenyl, alkanoylalkenyl, carboxyalkenyl Carbamoyl alkenyls; Substituted alkynyls having 1 to 10 carbon atoms, e.g., hydroxyalkynyls, alkoxyalkynyls, mercaptoalkynyls, alkylthioalkynyls, halogenoalkynyls, cyanoalkynyls, aminoalkynyls, alkylaminoalkynyls, dialkylaminoalkynyls, alkanoylalkynyls, carboxyalkynyls, carbamoylalkynyls; Aroylalkyls having up to 10 carbon atoms, e.g., phenacyl or 2-benzoylethyl; Aryls, e.g., phenyl or 1-naphthylethyl; Tyl; heteroaryl, e.g., 4-quinolyl; alkanoyl having 1 to 10 carbon atoms, e.g., acetyl or butyryl; aroyl, e.g., benzoyl; heteroaloyl, e.g., 3-quinoloyl; OR' or NR'R'' (where R' and R'' are independently hydrogen, alkyl, aryl, heteroaryl, acyl, aroyl, sulfonyl, or sulfinyl), SO2-R''' or SO-R''' (where R''' is substituted or unsubstituted alkyl, aryl, heteroaryl, alkenyl, or alkynyl).

[0178] In some embodiments, the capping functional group providing the ester bond is denoted as OR, where R can be an alkoxy; aryloxy; heteroaryloxy; aralkyloxy; heteroaralkyloxy; substituted alkoxy; substituted aryloxy; substituted heteroaryloxy; substituted aralkyloxy; or substituted heteroaralkyloxy.

[0179] In some embodiments, the peptides of the present disclosure may include modifications to the C-terminus of the peptide sequence by one or more of the following parts: NH2, NH-CH3; NH-CH2-CH3; NH-CH-(CH3)2, NH-CH2-CH2-CH3, NH-CH2-CH-(CH3)2, N(CH3)2, N(CH2-CH3)2, or OH.

[0180] In some embodiments, the peptides of the Disclosure may include modification of the N-terminus of a peptide sequence by one or more peptide-based moieties. In some embodiments, the peptides of the Disclosure may include modification of the C-terminus of a peptide sequence by one or more peptide-based moieties. In some embodiments, the peptides of the Disclosure may include modification of both the N-terminus and the C-terminus of a peptide sequence by one or more peptide-based moieties.

[0181] In some embodiments, the peptides of the Disclosure may include modification of the N-terminus of a peptide sequence by one or more non-peptide-based moieties. In some embodiments, the peptides of the Disclosure may include modification of the C-terminus of a peptide sequence by one or more non-peptide-based moieties. In some embodiments, the peptides of the Disclosure may include modification of both the N-terminus and the C-terminus of a peptide sequence by one or more non-peptide-based moieties.

[0182] In some embodiments, the peptides of the Disclosure may include N-terminal modifications comprising a series of 5 or 6 Glu amino acids. In some embodiments, the peptides of the Disclosure may include N-terminal modifications comprising a series of 5 or 6 Lys amino acids. In some embodiments, the peptides of the Disclosure may include N-terminal modifications comprising a series of 5 or 6 amino acids independently selected from Glu or Lys.

[0183] In some embodiments, the peptides of the Disclosure include an N-terminal peptide consisting of a chain of about 15 to about 400 identical amino acids. In some embodiments, the N-terminal peptide includes about 25 to about 300 identical amino acids, about 50 to about 200 identical amino acids, about 75 to about 150 identical amino acids, about 90 to about 120 identical amino acids, or about 100 or 110 identical amino acids. In some embodiments, the N-terminal peptide includes poly(glutamic acid) polypeptide (PGa), poly(aspartic acid) polypeptide (PA), poly(lysine) polypeptide (PLy), poly(arginine) polypeptide (PAr), poly(histidine) polypeptide (PHi), poly(ornithine) polypeptide (POr), or a combination thereof.

[0184] In some embodiments, the peptides of this disclosure include N-terminal modifications. In further embodiments, the N-terminal modification includes an N-terminal acetyl group (represented as Ac). For example, in SEQ ID NOs. 24-44 and 67-89 in Table 2, the N-terminal methionine group is capped with acetic anhydride or other suitable reagent during peptide synthesis to result in an N-terminally acetylated molecule. In some embodiments, the peptides of this disclosure include C-terminal modifications. In further embodiments, the C-terminal modification includes an amide group (represented as amide or CONH2). For example, in SEQ ID NOs. 24-44 and 67-89 in Table 2, the C-terminal serine group is synthesized as an amide during peptide synthesis to result in a C-terminally amidated molecule.

[0185] In some embodiments, the targeting moiety includes one or more peptide sequences that bind to DLL3 or a fragment or variant thereof, as listed in Table 2. In some embodiments, the targeting construct includes a targeting moiety that includes one or more peptide sequences that bind to DLL3 or a fragment or variant thereof, as listed in Table 2.

[0186] For example, the amino acid sequences of SEQ ID NOs: 3-24 are unmodified linear peptides that do not have a capping group before the formation of a disulfide bond. The amino acid sequences of SEQ ID NOs: 24-44 are linear peptides that have an acetyl group at the N-terminus and an amide group at the C-terminus before the formation of a disulfide bond. The amino acid sequences of SEQ ID NOs: 45-66 are unmodified cyclic peptides that do not have a capping group after the formation of a disulfide bond. The amino acid sequences of SEQ ID NOs: 67-89 are cyclic peptides that have an acetyl group at the N-terminus and an amide group at the C-terminus after the formation of a disulfide bond.

[0187] [Table 19]

[0188] [Table 20]

[0189] [Table 21]

[0190] The polypeptides of this disclosure may be peptide mimes. A "peptide mime" or "polypeptide mime" is a polypeptide whose molecule contains structural elements not found in natural polypeptides (i.e., polypeptides composed only of 20 protein-forming amino acids). In some embodiments, peptide mimes can reproduce or mimic the biological effects of natural peptides. Peptide mimes may differ in many ways from natural polypeptides, for example, by alterations in their skeletal structure or the presence of amino acids that do not exist naturally. In some cases, the peptide mimetic may include amino acids having side chains not found among the 20 known proteinogenic amino acids; non-polypeptide-based crosslinking moieties used to bring about cyclization between the terminal or internal parts of the molecule; substitution of amide bond hydrogens by methyl groups (N-methylation) or other alkyl groups; substitution of amino acid α-hydrogens by methyl groups (α-methylation) or substitution by other alkyl groups; substitution of peptide bonds by chemical groups or bonds resistant to chemical or enzymatic treatment; N-terminal and C-terminal modifications; and / or conjugations with non-peptide extensions (e.g., polyethylene glycol, lipids, carbohydrates, nucleosides, nucleotides, nucleoside bases, various small molecules, or phosphate or sulfate groups).

[0191] As used herein, the term “amino acid” includes residues of both natural and non-natural amino acids. The 20 natural protein-forming amino acids are identified and referenced herein by either one-letter or three-letter notation as follows: aspartic acid (Asp:D), isoleucine (Ile:I), threonine (Thr:T), leucine (Leu:L), serine (Ser:S), tyrosine (Tyr:Y), glutamic acid (Glu:E), phenylalanine (Phe:F), proline (Pro:P), histidine (His:H), glycine (Gly:G), lysine (Lys:K), alanine (Ala:A), arginine (Arg:R), cysteine ​​(Cys:C), tryptophan (Trp:W), valine (Val:V), glutamine (Gln:Q), methionine (Met:M), and asparagine (Asn:N). Natural amino acids exist in their levorotatory (L) stereoisomer forms. Unless otherwise specified, the amino acids referred to herein are L-stereoisomers. The term “amino acid” includes amino acids having conventional amino protecting groups (e.g., acetyl or benzyloxycarbonyl), as well as natural and unnatural amino acids protected at the carboxyl terminus (e.g., as (C1-C6) alkyl, phenyl, or benzyl esters or amides; or as α-methylbenzylamides). Other suitable amino and carboxyl protecting groups are known to those skilled in the art (see, for example, Greene, TW; Wutz, PGM, Protecting Groups In Organic Synthesis; second edition, 1991, New York, John Wiley & Sons, Inc., whose entire contents are incorporated herein by reference, and the literature cited therein). The peptides and / or peptide compositions of this disclosure may also contain modified amino acids.

[0192] "Non-natural" amino acids have side chains or other features not present in the 20 naturally occurring amino acids listed above, and include, but are not limited to, N-methyl amino acids, N-alkyl amino acids, α,α-substituted amino acids, β-amino acids, α-hydroxy amino acids, D-amino acids, and other non-natural amino acids known in the art (e.g., Josephson et al., (2005) J.Am. Chem. Soc. 127:11727-11735; Forster, AC et al. (2003) Proc. Natl. Acad. Sci. USA 100:6353-6357; Subtelny et al., (2008) J.Am. Chem. Soc. 130:6131-6136; Hartman, MCT et al. (2007) PLoS ONE 2:e972; and Hartman et al. See al., (2006) Proc. Natl. Acad. Sci. USA 103:4356-4361). Further non-natural amino acids useful for optimizing the peptides and / or peptide compositions of this disclosure include, but are not limited to, 1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid, 1-amino-2,3-hydro-1H-indene-1-carboxylic acid, homolysine, homoarginine, homoserine, 2-aminoadipic acid, 3-aminoadipic acid, β-alanine, aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 5-aminopentanoic acid, 5-aminohexanoic acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, desmosine, 2,3-Diaminopropionic acid, N-ethylglycine, N-ethylasparagine, homoproline, hydroxylysine, allo-hydroxylysine, 3-hydroxyproline, 4-hydroxyproline, isodesmosine, allo-isoleucine, N-methylpentylglycine, naphthylalanine, ornithine, pentylglycine, thioproline, norvaline, tert-butylglycine (also known as tert-leucine), phenylglycine, azatryptophan, 5-azatryptophan, 7-azatryptophan, 4-fluorophenylalanine, penicillamine, sarcosine, homocysteine, 1-aminocyclopropanecarboxylic acid, 1-aminocyclobutanecarboxylic acid, 1-aminocyclopentanecarboxylic acid, 1-aminocyclohexanecarboxylic acid, 4-aminotetrahydro-2H-pyran-4-carboxylic acid, (S)-2-amino-3-(1H-tetra Razole-5-yl)propanoic acid, cyclopentylglycine, cyclohexylglycine, cyclopropylglycine, η-ω-methylarginine, 4-chlorophenylalanine, 3-chlorotyrosine, 3-fluorotyrosine, 5-fluorotryptophan, 5-chlorotryptophan, citrulline, 4-chloro-homophenylalanine, homophenylalanine, 4-aminomethylphenylalanine, 3-aminomethylphenylalanine, octylglycine, norleucine, tranexamic acid, 2-aminopentanoic acid, 2-aminohexanoic acid, 2-aminoheptanoic acid, 2-aminooctanoic acid, 2-aminononanoic acid, 2-aminodecanoic acid, 2-aminoundecanoic acid, 2-aminododecanoic acid, aminovaleric acid, and 2-(2-aminoethoxy)acetic acid, pipecoric acid, 2-carboxyazetidine, hexafluoroleucine, 3-fluorovaline, 2-amino-4,4-Difluoro-3-methylbutanoic acid, 3-fluoroisoleucine, 4-fluoroisoleucine, 5-fluoroisoleucine, 4-methylphenylglycine, 4-ethylphenylglycine, 4-isopropylphenylglycine, (S)-2-amino-5-azidopentaic acid (also referred to herein as "X02"), (S)-2-aminohepta-6-enoic acid (also referred to herein as "X30"), (S)-2-aminopenta-4-icic acid (also referred to herein as "X31") (Also referred to as "X12"), (S)-2-aminopenta-4-enoic acid (also referred to as "X12" herein), (S)-2-amino-5-(3-methylguanidino)pentanoic acid, (S)-2-amino-3-(4-(aminomethyl)phenyl)propanoic acid, (S)-2-amino-3-(3-(aminomethyl)phenyl)propanoic acid, (S)-2-amino-4-(2-aminobenzo[d]oxazole-5-yl)butanoic acid, (S)-leucinol, (S)-valinol, (S)-te rt-leucinol, (R)-3-methylbutan-2-amine, (S)-2-methyl-1-phenylpropane-1-amine, and (S)-N,2-dimethyl-1-(pyridine-2-yl)propane-1-amine, (S)-2-amino-3-(oxazole-2-yl)propanoic acid, (S)-2-amino-3-(oxazole-5-yl)propanoic acid, (S)-2-amino-3-(1,3,4-oxadiazole-2-yl)propanoic acid, (S)-2-amino-3-(1,2, 4-Oxadiazole-3-yl)propanoic acid, (S)-2-amino-3-(5-fluoro-1H-indazole-3-yl)propanoic acid, and (S)-2-amino-3-(1H-indazole-3-yl)propanoic acid, (S)-2-amino-3-(oxazol-2-yl)butanoic acid, (S)-2-amino-3-(oxazol-5-yl)butanoic acid, (S)-2-amino-3-(1,3,4-oxadiazole-2-yl)butanoic acid, (S)-2-amino-3-(1,2,Examples include 4-oxadiazole-3-yl)butanoic acid, (S)-2-amino-3-(5-fluoro-1H-indazole-3-yl)butanoic acid, and (S)-2-amino-3-(1H-indazole-3-yl)butanoic acid, 2-(2'MeOphenyl)-2-aminoacetic acid, tetrahydro-3-isoquinoline carboxylic acid, and their stereoisomers (including, but not limited to, D and L isomers).

[0193] Further non-natural amino acids useful for optimizing the peptides or peptide compositions of this disclosure include, but are not limited to, halogenated amino acids in which one or more carbon-bonded hydrogen atoms are substituted with one or more halogen atoms. The number of halogen atoms may range from one to all hydrogen atoms.

[0194] In some embodiments, non-natural amino acids useful for optimizing the peptides or peptide compositions of the present disclosure include, but are not limited to, fluorinated amino acids in which one or more carbon-bonded hydrogen atoms are substituted with one or more fluorine atoms. The number of fluorine atoms may range from one to all hydrogen atoms. Examples of such amino acids include, but are not limited to, 3-fluoroproline, 3,3-difluoroproline, 4-fluoroproline, 4,4-difluoroproline, 3,4-difluoroproline, 3,3,4,4-tetrafluoroproline, 4-fluorotryptophan, 5-fluorotryptophan, 6-fluorotryptophan, 7-fluorotryptophan, and their stereoisomers.

[0195] In some embodiments, but not limited to them, non-natural amino acids useful for optimizing the peptides or peptide compositions of this disclosure include chlorinated amino acids in which one or more carbon-bonded hydrogen atoms are substituted with one or more chlorine atoms. The number of chlorine atoms may range from one to all hydrogen atoms.

[0196] Further non-natural amino acids useful for optimizing the peptides of this disclosure include, but are not limited to, those disubstituted at the α-carbon. These include amino acids with the same two substituents on the α-carbon, such as α-aminoisobutyric acid and 2-amino-2-ethylbutanoic acid, and amino acids with different substituents, such as α-methylphenylglycine and α-methylproline. Furthermore, the substituents on the α-carbon may together form rings, such as 1-aminocyclopentanecarboxylic acid, 1-aminocyclobutanecarboxylic acid, 1-aminocyclohexanecarboxylic acid, 3-aminotetrahydrofuran-3-carboxylic acid, 3-aminotetrahydropyran-3-carboxylic acid, 4-aminotetrahydropyran-4-carboxylic acid, 3-aminopyrrolidine-3-carboxylic acid, 3-aminopiperidine-3-carboxylic acid, 4-aminopiperidine-4-carboxylic acid, and their stereoisomers.

[0197] Further non-natural amino acids useful for optimizing the peptides or peptide compositions of this disclosure include, but are not limited to, tryptophan analogs in which the indole ring system is substituted by another 9 or 10-membered bicyclic ring system containing 0, 1, 2, 3, or 4 heteroatoms independently selected from N, O, or S. Each ring system may be saturated, partially unsaturated, or fully unsaturated. The ring systems may be substituted with 0, 1, 2, 3, or 4 substituents of any substituteable atom. Each substituent may be independently selected from H, F, Cl, Br, CN, COOR, CONRR', oxo, OR, NRR'. Each R and R' may be independently selected from H, C1-C20 alkyl, or C1-C20 alkyl-O-C1-20 alkyl.

[0198] In some embodiments, tryptophan analogs (also referred to herein as “tryptophan analogs”) may be useful for optimizing the peptides or peptide compositions of this disclosure. Examples of tryptophan analogs include, but are not limited to, 5-fluorotryptophan [(5-F)W], 5-methyl-O-tryptophan [(5-MeO)W], 1-methyltryptophan [(1-Me-W) or (1-Me)W], D-tryptophan (D-Trp), azatryptophan (including, but not limited to, 4-azatryptophan, 7-azatryptophan, and 5-azatryptophan), 5-chlorotryptophan, 4-fluorotryptophan, 6-fluorotryptophan, 7-fluorotryptophan, and their stereoisomers. Unless otherwise stated, the term “azatryptophan” and its abbreviation “azaTrp” refer to 7-azatryptophan as used herein.

[0199] Modified amino acid residues useful for optimizing the peptides and / or peptide compositions of this disclosure include, but are not limited to, those that are chemically blocked (reversibly or irreversibly); those whose N-terminal amino group or their side chain groups are chemically modified; those whose amide skeletons are chemically modified, such as N-methylation, D (non-natural amino acid) and L (natural amino acid) stereoisomers; or residues whose side chain functional groups are chemically modified to other functional groups. In some embodiments, modified amino acids include, but are not limited to, methionine sulfoxides; methionine sulfones; aspartate-(β-methyl ester), modified amino acids of aspartate; N-ethylglycine, modified amino acids of glycine; alanine carboxamide; and / or modified amino acids of alanine. Non-natural amino acids may be purchased from Sigma-Aldrich (St. Louis, MO), Bachem (Torrance, CA) or other suppliers. Non-natural amino acids may further include any of those listed in Table 2 of U.S. Patent Application Publication No. 2011 / 0172126, the entirety of which is incorporated herein by reference.

[0200] In some embodiments, the amino acids for use in this disclosure are modified using organic protein or non-protein derivative agents. In some embodiments, the amino acids for use in this disclosure are modified using post-translational modifications. In some embodiments, the modification is introduced by reacting a targeted amino acid residue of the peptide with an organic derivative agent that can react with a selected side-chain or terminal residue. In some embodiments, the modification is introduced by utilizing a post-translational modification mechanism that functions in a selected recombinant host cell. Specific post-translational modifications are the result of the recombinant host cell's action on the expressed peptide. As an example, glutaminyl and asparaginyl residues are often deamidated under specific post-translational conditions (e.g., weakly acidic conditions) to become the corresponding glutamyl and aspartyl residues. Other post-translational modifications include hydroxylation of proline and lysine; phosphorylation of hydroxyl groups of tyrosinyl, ceryl, or threonyl residues; and methylation of α-amino groups of lysine, arginine, and histidine side chains (e.g., Creighton et al., Proteins: Structure and Molecular Properties, WH Freeman & Co., San Francisco, 1983, pp. 79-86).

[0201] In some embodiments, amino acid modifications include the binding of non-proteinaceous polymers to the peptides of this disclosure. Examples of non-proteinaceous polymers include hydrophilic synthetic polymers (i.e., non-natural polymers) such as hydrophilic polyvinyl polymers (e.g., polyvinyl alcohol and polyvinylpyrrolidone). Examples of non-proteinaceous polymers also include polyethylene glycol, polypropylene glycol, and polyoxyalkylenes. In some embodiments, amino acid modifications include the binding of non-proteinaceous polymers to the peptides of this disclosure, as described in U.S. Patent No. 4,640835, U.S. Patent No. 4,496689, U.S. Patent No. 4,301144, U.S. Patent No. 4,670417, U.S. Patent No. 4,791192, and U.S. Patent No. 4,179337 (the contents of which are each incorporated herein by reference in whole in relation to the amino acid modifications of this disclosure).

[0202] Peptide synthesis This disclosure provides methods for synthesizing the peptides and compounds of this disclosure. In some embodiments, the peptides of this disclosure can be obtained by inducing the formation of a covalent bond between the N-terminal amino group (if provided) of the peptide and the carboxyl group of a reactive amino acid side chain (if provided). In some embodiments, the peptides and compounds of this disclosure can be synthesized by any known conventional procedure for the formation of peptide bonds between amino acids. Such conventional procedures include, for example, any solution-phase procedure that enables the condensation between a free α-amino group of an amino acid or residue thereof (whose carboxyl group or other reactive group is protected) and a free primary carboxyl group of another amino acid or residue thereof (whose amino group or other reactive group is protected). In some embodiments, the peptides of this disclosure can be synthesized by solid-phase synthesis and purified according to methods known in the art. The peptides of this disclosure can be prepared using any of several well-known procedures utilizing a variety of resins and reagents.

[0203] In some embodiments, a method for synthesizing a peptide may be carried out by a procedure in which each amino acid in a desired sequence is successively added one at a time to another amino acid or residue thereof. In some embodiments, a method for synthesizing a peptide may be carried out by a procedure in which a plurality of peptide fragments having a portion of a desired amino acid sequence are first synthesized and then condensed to provide the desired peptide sequence.

[0204] In some embodiments, the method for synthesizing the peptide can be carried out using solid-phase peptide synthesis, which includes methods well known and practiced in the art (e.g., the Symphony Multiplex Peptide Synthesizer (Rainin Instrument Company) automated peptide synthesizer). In some embodiments, the method for synthesizing the peptide can be carried out using a standard Fmoc method on an automated synthesizer (e.g., Advanced ChemTech 440M05, Louisville, Ky). In some embodiments, the method for synthesizing the peptide can be carried out using a coupling reagent such as 2-(1-H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU) and / or 1-hydroxybenzotriazole (HOBt).

[0205] Solid-phase peptide synthesis can be carried out by sequentially incorporating the desired amino acid residues one at a time into the growing side chain, in accordance with the general principles of solid-phase synthesis. These methods are disclosed in numerous references, including Merrifield, et al., Solid phase synthesis (Nobel lecture), Angew, Chem. (1985) 24:799-810; Barany et al., The Peptides, Analysis, Synthesis and Biology, Vol.2; and Gross et al., Eds. Academic Press 1-284 (1980), the contents of which, in relation to methods and protocols for peptide synthesis, are incorporated herein by reference in their entirety.

[0206] Solid-phase synthesis of peptides generally begins from the C-terminus of the peptide by coupling a protected α-amino acid to a suitable resin. Examples of known methods for preparing substituted amide derivatives on a solid phase have been described in the Art (see, for example, Barn D et al., Tetrahedron Letters (1996), 37:3213-3216; DeGrado et al., J. Org. Chem., (1982) 47:3258-3261; the contents of these, respectively, relating to methods and systems for solid-phase peptide synthesis, are incorporated herein by reference in their entirety). As an example, the starting material can be prepared by known means by esterifying an α-aminoprotected amino acid to a p-benzyloxybenzyl alcohol (Wang) resin or oxime resin. The peptide chain is grown with the desired amino acid sequence, and then the peptide-resin is treated with a solution of a suitable amine (such as methylamine, dimethylamine, or ethylamine). Peptides using p-benzyloxybenzyl alcohol (Wang) resin can be cleaved from the resin by aluminum chloride in DCM, while peptides using oxime resin can be cleaved by DCM.

[0207] In some embodiments, the reactive side chain groups of various amino acid residues are protected with a suitable protecting group to prevent chemical reactions from occurring at that site until the protecting group is removed. In some embodiments, the α-amino group of an amino acid residue or fragment is protected, while the entity reacts with a carboxyl group, after which the α-amino protecting group can be selectively removed to allow subsequent reactions at that site. Examples of protecting groups for use in this disclosure are disclosed and are known in solid-phase and liquid-phase synthesis.

[0208] In some embodiments, the α-amino group can be protected by urethane protecting groups, such as benzyloxycarbonyl (Z) and substituted benzyloxycarbonyl groups, such as p-chlorobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, p-biphenyl-isopropoxycarbonyl, 9-fluorenylmethoxycarbonyl (Fmoc) and p-methoxybenzyloxycarbonyl (Moz); and aliphatic urethane protecting groups, such as t-butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl, isopropoxycarbonyl, and allyloxycarbonyl.

[0209] In some embodiments, the guanidinoamino group (such as that found in arginine) can be protected by suitable protecting groups, such as nitro, p-toluenesulfonyl (ToS), Z, pentamethylchromansulfonyl (Pmc), adamantyloxycarbonyl, pentamethyldihydrobenzofuran-5-sulfonyl (Pbf), and Boc.

[0210] As a non-limiting example, solid-phase synthesis of peptides can be initiated from the C-terminus of a peptide by coupling a protected α-amino acid to a suitable resin. Starting materials can be prepared by esterifying the α-amino-protected amino acid to a p-benzyloxybenzyl alcohol (Wang) resin, a 2-chlorotrityl chloride resin, or an oxime resin, by amide bonding between an Fmoc-linker, e.g., p-[(R,S)-α-[1-(9H-fluoro-en-9-yl)-methoxyformamide]-2,4-dimethyloxybenzyl]-phenoxyacetic acid (Rink linker), or by other means well known in the art. Fmoc-linker-BHA resin supports are commercially available and commonly used where possible. The resin is then subjected to iterative addition cycles as needed to sequentially add amino acids. Next, the α-aminoFmoc protecting group is removed under basic conditions (e.g., piperidine, piperazine, diethylamine, or morpholine (20-40% v / v) in N,N-dimethylformamide (DMF)). After the removal of the α-amino protecting group, the subsequent protected amino acids are coupled stepwise in the desired order to obtain an intermediate protected peptide-resin. Activating reagents used for amino acid coupling in solid-phase peptide synthesis are well known in the art. After the peptide is synthesized, the orthogonally protected side chain protecting group can be removed, if necessary, using methods well known in the art for further derivatization of the peptide.

[0211] Reactive groups in peptides can be selectively modified either during solid-phase synthesis or after removal from the resin. For example, peptides can be modified on the resin to obtain N-terminal modifications such as acetylation, or they can be removed from the resin using cleavage reagents and then modified. Similarly, methods for modifying amino acid side chains are well known to those skilled in peptide synthesis. The choice of modifications to be made to reactive groups present on the peptide will be determined in part by the desired properties of the peptide.

[0212] In some embodiments, the N-terminal group is modified by the introduction of an N-acetyl group. In non-limiting examples, peptide synthesis may include the step of reacting a resin-bound peptide with acetic anhydride in dichloromethane in the presence of an organic base such as diisopropylethylamine, after removal of the N-terminal protecting group. Other methods of N-terminal acetylation, including solution-phase acetylation, are known in the art.

[0213] In some embodiments, the peptides of the present disclosure may include cyclic peptides having one or more crosslinking sites (e.g., cyclic structures, staples, crosslinks, etc.).

[0214] In some embodiments, peptides may be synthesized using solid-phase peptide synthesis and then cyclized before being cleaved from the peptide resin. When peptides are cyclized via a reactive side chain moiety, the desired side chain is first deprotected in a suitable solvent under specific deprotection conditions, and then a cyclic coupling agent is added. Suitable solvents include, but are not limited to, DMF, dichloromethane (DCM), and 1-methyl-2-pyrrolidone (NMP). Suitable cyclic coupling reagents include, but are not limited to, 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), benzotriazole-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP), and benzotriazole-1-yl- Examples include xytris(pyrrolidino)phosphonium hexafluorophosphate (PyBOP), 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TATU), 2-(2-oxo-1(2H)-pyridyl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TPTU), and N,N'-dicyclohexylcarbodiimide / 1-hydroxybenzotriazole (DCCl / HOBt). In some embodiments, coupling of the cyclic moiety with the peptide chain is initiated by the use of a suitable base such as N,N-diisopropylethylamine (DIPEA), sym-collidine, or N-methylmorpholine (NMM).

[0215] Next, the cyclized peptide can be cleaved from the solid phase using any suitable reagent, such as ethylamine in DCM. The resulting crude peptide is dried, and any remaining amino acid side-chain protecting groups (if present) are cleaved using a suitable reagent, such as trifluoroacetic acid (TFA), in the presence of water and 1,2-ethanedithiol (EDT). The final product is precipitated by adding cold ether and recovered by filtration. Final purification may be performed by reversed-phase high-performance liquid chromatography (RP-HPLC) using a suitable column, such as a C18 column. Other methods of separation or purification, such as those based on peptide size or charge, may also be used. Once purified, the peptide can be characterized by several methods, including high-performance liquid chromatography (HPLC), amino acid analysis, and mass spectrometry.

[0216] In some embodiments, the peptides of this disclosure may include one or more modifications (e.g., substitution, addition, or deletion) to one or more ends of the peptide sequence (e.g., N-terminus, C-terminus, or both). In some embodiments, the terminally modified peptides may be synthesized using solid-phase peptide synthesis and modified before being cleaved from the peptide resin.

[0217] This disclosure intends to describe variants and derivatives of the peptides described herein. These include substitutional, insertional, deletional, and covalent variants and derivatives. As used herein, the term “derivative” is used synonymously with the term “variant” and refers to a molecule that has been modified or altered in any way relative to the reference molecule or the starting molecule.

[0218] In one embodiment, the peptide described herein includes the substitution of one or more L-amino acid residues with one or more D-amino acid residues. This embodiment is thought to increase proteolytic stability due to steric hindrance and the tendency of D-amino acids to stabilize the β-turn conformation (Tugyi et al (2005) PNAS, 102(2), 413-418).

[0219] In some embodiments, the peptides of this disclosure may be in salt form. Salts of peptides can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods, such as those described in Pharmaceutical Salts: Properties, Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002. Generally, such salts can be prepared by reacting the free acidic or basic form of these compounds with a suitable base or acid in water, an organic solvent, or a mixture thereof.

[0220] Acid addition salts (mono or disal) can be formed with a wide variety of both inorganic and organic acids. Examples of acid addition salts include acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid (e.g., L-ascorbic acid), L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, butyric acid, (+)camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, and cyclamate. Lamin acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid, glucuronic acid (e.g., D-glucuronic acid), glutamic acid (e.g., L-glutamic acid), α-oxoglutaric acid, glycolic acid, hippuric acid, hydrohalogens (e.g., hydrobromic acid, hydrochloric acid) Hydroiodic acid, isethionic acid, lactic acid (e.g., (+)-L-lactic acid, (±)-DL-lactic acid), lactobionic acid, maleic acid, malic acid, (-)-L-malic acid, malonic acid, (±)-DL-mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid Examples include monosal or disalts formed with acids selected from the group consisting of nic acid, pamoic acid, phosphoric acid, propionic acid, pyruvic acid, L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, undecylenic acid, and valeric acid, as well as acylated amino acids and cation exchange resins.

[0221] In some embodiments, the salts of the present disclosure may be salts formed from acetic acid, hydrochloric acid, hydroiodic acid, phosphoric acid, nitric acid, sulfuric acid, citric acid, lactic acid, succinic acid, maleic acid, malic acid, isethionic acid, fumaric acid, benzenesulfonic acid, toluenesulfonic acid, sulfuric acid, methanesulfonic acid (mesylate), ethanesulfonic acid, naphthalenesulfonic acid, valeric acid, propanoic acid, butanoic acid, malonic acid, glucuronic acid, and lactobionic acid. In some embodiments, the salt may be a hydrochloride salt. In some embodiments, the salt may be an acetate salt.

[0222] If a compound is anionic or has a functional group that can be anionic (for example, -COOH may be -COO-), the salt can be formed with an organic or inorganic base to produce a suitable cation. Examples of suitable inorganic cations include, but are not limited to, Li + na + and K + Alkali metal ions such as Ca 2+ and Mg 2+ Examples include alkaline earth metal cations such as NH4, as well as other cations. Suitable organic cations include, but are not limited to, ammonium ions (i.e., NH4) + ) and substituted ammonium ions (e.g., NH3R + NH2R 2+ NHR 3+ and NR 4+ Examples of suitable substituted ammonium ions include those derived from methylamine, ethylamine, diethylamine, propylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, and tromethamine, as well as amino acids such as lysine and arginine. A common example of a quaternary ammonium ion is N(CH3)4 + That is the case.

[0223] If the peptides contain amine functional groups, they can form quaternary ammonium salts, for example, by reaction with alkylating agents, according to methods well known to those skilled in the art.

[0224] The peptides disclosed herein have an equilibrium dissociation constant (K) of approximately 0.001 nM to approximately 0.01 nM, approximately 0.005 nM to approximately 0.05 nM, approximately 0.01 nM to approximately 0.1 nM, approximately 0.05 nM to approximately 0.5 nM, approximately 0.1 nM to approximately 1.0 nM, approximately 0.5 nM to approximately 5.0 nM, approximately 2 nM to approximately 10 nM, approximately 8 nM to approximately 20 nM, approximately 15 nM to approximately 45 nM, approximately 30 nM to approximately 60 nM, approximately 40 nM to approximately 80 nM, approximately 50 nM to approximately 100 nM, approximately 75 nM to approximately 150 nM, approximately 100 nM to approximately 500 nM, approximately 200 nM to approximately 800 nM, approximately 400 nM to approximately 1,000 nM, or at least 1,000 nM. D ) can bind to the target receptor.

[0225] In some embodiments, the peptides disclosed herein have an equilibrium dissociation constant (K) of approximately 0.001 nM to approximately 0.01 nM, approximately 0.005 nM to approximately 0.05 nM, approximately 0.01 nM to approximately 0.1 nM, approximately 0.05 nM to approximately 0.5 nM, approximately 0.1 nM to approximately 1.0 nM, approximately 0.5 nM to approximately 5.0 nM, approximately 2 nM to approximately 10 nM, approximately 8 nM to approximately 20 nM, approximately 15 nM to approximately 45 nM, approximately 30 nM to approximately 60 nM, approximately 40 nM to approximately 80 nM, approximately 50 nM to approximately 100 nM, approximately 75 nM to approximately 150 nM, approximately 100 nM to approximately 500 nM, approximately 200 nM to approximately 800 nM, approximately 400 nM to approximately 1,000 nM, or at least 1,000 nM. D ) can be coupled to DLL3.

[0226] Chelating agents The chelating agent (CA) (also referred to herein as “chelating agent”) may include a metal chelating agent that associates with a metal cargo (e.g., a metal nuclide cargo). The chelating agent may include a polymer compound. In some embodiments, the chelating agent includes an acyclic or macrocyclic compound.

[0227] Examples of chelating agents (also called "chelating agents") are shown in Table C below.

[0228] Table 22

[0229] Table 23

[0230] Table 24

[0231] Table 25

[0232] Table 26

[0233] Table 27

[0234] Table 28

[0235] Table 29

[0236] Table 30

[0237] In some embodiments, the chelating agent includes acyclic or macrocyclic compounds. Non-limiting examples of chelating agents include 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA); DOTA derivative: DO3A; diethylenetriamine-N,N,N',N”,N”-pentaacetic acid (DTPA); DTPA derivatives: 2-(p-SCN-Bz)-”methyl-DTPA, CHX-A”-DTPA, and cyclic anhydride of DTPA (CA-DTPA); 1,4,7-triazacyclononane-1,4-7-triacetic acid (NOTA); NOTA derivatives (e.g., BCNOTA, p-NCS-Bz-NOTA, BCNOT); 6-hydrazinonicotinamide HYNIC; ethylenediaminetetraacetic acid (EDTA); N,N'-ethylene-di-L-cysteine; N,N'-bis(2,2-dimethyl-2-mercaptoethyl)ethylenediamine-N,N'-diacetic acid (6SS); 1-(4-carboxymethoxybenzyl)-N-N'-bis[(2-mercapto-2,2-dimethyl)ethyl]-1,2'ethylenediamine-N,N'-diacetic acid (B6SS); deferoxamine (DFO); 1,1,1-tris(aminomethyl)ethane (TAME); tris(aminomethyl)ethane-N,N,N',N',N”,N”-hexaacetic acid (TAME Hex); O-Hydroxybenzyliminodiacetic acid; 1,4,7-Triazacyclononane (TACN); 1,4,7,10-Tetraazacyclododecane (Cyclene); 1,4,7-Triazacyclononane-1-succinate-4,7-diacetic acid (NODASA); 1-(1-carboxy-3-carboxypropyl)-4,7-bis-(carboxymethyl)-1,4,7-triazacyclononane (NODAGA); 1,4,7-Tris(2-mercaptoethyl)-1,4,7-triazacyclononane ( Triazacyclononane™; 1,4,7-Triazacyclononane-N,N',N”-Tris(methylenephosphonic acid) (NOTP); 1,4,8,11-Tetraazacyclotetradecane-N,N',N”,N”'-Tetraacetic acid (TETA); 1,4,7,10,13-Pentazacyclopentadecane-N,N',N”,N”',N””-Pentaacetic acid (PEPA), 1,4,7,10,13,16-Hexaazacyclohexadecane-N,N',N”,N”',N””,N””'-Hexaacetic acid (HEHA);Examples include 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane (TCMC) and its derivatives or analogues.

[0238] In one embodiment, the chelating agent is independently ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 1,4,7,10-tetra-azacyclododecane-N,N',N”,N”'-tetraacetic acid (DOTA), 6-((16-((6-carboxypyridine-2-yl)methyl)-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane-7-yl)methyl)-4-isothiocyanatopicolinic acid (Macropa), Macrodipa, 2,2',2”,2”'-(1,10-dioxa-4,7,13,16-tetraazacyclooctadecane-4,7,13,16-tetra Selected from the group consisting of (Tryl)tetraacetic acid (Crown), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, α-(2-carboxyethyl) (DOTAGA), 1,4,7-triazacyclononane-N,N',N''-triacetic acid (NOTA), 1,4,7,10-tetraazacyclododecane-N,N',N",N'''-tetraacetic acid (TETA), 1,4,7,10,13-pentazacyclopentadecane-N,N',N",N''',N'''-pentaacetic acid (PEPA), and 1,4,7,10,13,16-hexaazacyclohexadecane-N,N',N",N''',N''",N'''-hexaacetic acid (HEHA).

[0239] In some embodiments, the chelating agents of this disclosure include DOTA, DOTAGA, or any derivatives / analogs thereof. Any chelating agent disclosed in Eisenwiener et al., Bioorg Med Chem Lett., vol.10(18):2133(2000) (the contents of which are incorporated herein by reference in their entirety) may be used as a chelating agent.

[0240] Chelating agents such as DOTA can be bound to any position on a cyclic peptide without adversely affecting its binding to its target (i.e., those skilled in the art will be able to understand how the placement of the chelating agent affects binding by performing the tests described herein). In some embodiments, chelating agents such as DOTA can be bound directly to the N-terminal amine or to a short linker attached to the same residue. Alternatively, chelating agents such as DOTA can be bound to the C-terminus or side chain, which may tolerate their presence, via a short linker. In some embodiments, crosslinking agents such as dibromoxylen, which are pre-functionalized in the chelating moiety, can be bound to the cyclic peptide.

[0241] Linker The targeted construct may include an optional linker that links a chelating agent to a targeted moiety. The targeted construct linker may link one or more chelating agents to one or more targeted moieties. The linker may include one or more of the following: esters, disulfides, amides, acylhydrazones, ethers, carbamates, carbonates, sulfonamides, alkyls, aryls, heteroaryls, thioethers, and ureas.

[0242] In some embodiments, the linker includes a cleavable linker. In some embodiments, the linker includes an incleavable linker. In some embodiments, an optional linker includes an amino acid.

[0243] As used herein, the term “linker” refers to the chemical portion that links a chelating agent to the peptide of this disclosure. Any suitable linker known to those skilled in the art in consideration of this disclosure may be used herein.

[0244] The linker can act as an electrophile and bind to the nucleophilic moiety of the chelating agent. Alternatively, the linker can act as a nucleophile and bind to the electrophilic moiety of the chelating agent. It is understood that the linker can bind to the chelating agent via its carbon skeleton, allowing all "binding arms" of the chelating agent molecule to interact with the metal. Alternatively, one of the arms can bind to the linker.

[0245] For example, if a chelating agent is bonded to the carbonyl group of the chelating agent via an amine group of a cyclic peptide or an optional linker, an amide bond is formed between the chelating agent and the cyclic peptide or optional linker.

[0246] In another example, if the chelating agent is DOTA and the linker is PEG, the resulting structure is: [ka] It is possible.

[0247] In yet another example, if the chelating agent is DOTA and the amino acid is Lys, the resulting side chain of the amino acid is: [ka] It is possible.

[0248] cargo The targeted construct may contain various cargoes. In some embodiments, cargo association with the targeted construct is facilitated by a chelating agent. The cargo may contain a radioactive agent. A radioactive agent cargo that associates with the targeted construct via a chelating agent may contain radionuclides and / or radioisotopes. The chelating agent used for the association of such cargo with the targeted construct may contain a metal chelating group. In one embodiment, the chelating agent of the compound provided herein further contains a radioactive metal ion bonded to the chelating agent via a coordination bond, thereby forming a radioactive metal complex.

[0249] Various radionuclides possess radioactive properties including alpha, beta, gamma, and Auger radiation. These can be used in conjunction with targeted constructs for therapeutic and / or diagnostic purposes. For example, the activator Z may contain the radioactive isotopes Y-90, Y-86, I-131, Re-186, Re-188, Y-90, Bi-212, At-211, Zr-89, Sr-89, Ho-166, Sm-153, Cu-67, Cu-64, Lu-177, Ac-225, Pb-203, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc-47, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, Ir-194 and / or Pt-199.

[0250] In some embodiments, targeted constructs used for imaging applications may include radioisotope cargo useful as imaging probes. Non-limiting examples of such radioisotopes include, but are not limited to, I-124, I-131, In-111, Re-186, Re-188, Y-90, Bi-212, At-211, Sr-89, Ho-166, Sm-153, Cu-60, Cu-67, Cu-64, Lu-177, Ac-225, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc- Examples include 47, Br-76, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, In-111, Ir-194, Pt-199, Tc-99m, Co-57, Ga-66, Ga-67, Ga-68, Kr-81m, Rb-82, Sr-92, Tl-201, Y-86, Zr-89, C-11, N-13, O-15, and F-18.

[0251] In some embodiments, the targeted construct cargo includes any of the radioisotopes listed in Table 3, including the parent and daughter radionuclides.

[0252] [Table 31]

[0253] In some embodiments, the radionuclide is a therapeutically active radionuclide. Suitable therapeutically active radionuclides are, but are not limited to, 32 P, 67 Cu, 186 Re, 188 Re, 89 Sr, 90 Y, 143 Ce, 177 Lu, 161 Tb, 166 Ho, 169 Er, 183 Ta, 153 Sm, 213 Bi, 131 I, 149 Tb, 47 Sc, 225 Ac, 212 Pb, 211 At, 223 Ra, 227 Th, and 226 Th is one example. In some embodiments, the radionuclide is 67 Cu, 188 Re, 90 Y, 177 Lu, 213 Bi, 131 I, 47 Sc, 225 Ac, 212 Pb, 211 At, and 227 A therapeutically active radionuclide selected from Th. In certain embodiments, the radionuclide is 90 Y, 177 Lu, 131 I, 225 Ac, 211 At, and 227 A therapeutically active radionuclide selected from Th. In certain embodiments, the therapeutically active radionuclide is 177 It is Lu. In certain embodiments, the therapeutically active radionuclide is 225 It is Ac.

[0254] Alternatively, in some embodiments, the radionuclide is a diagnostically active radionuclide. Suitable diagnostically active radionuclides include, but are not limited to, 111 In, 99m Tc,94m Tc, 67 Ga, 68 Ga, 203 Pb, 64 Cu, 86 Y, 89 Zr, 51 Mn, 52 Mn, 123 I, 124 I, 125 I, 18 F, 76 Br, 77 Br, 152 Tb, 155 Tb, 44 Sc, 43 Sc, and 201 Tl is one example. In some embodiments, the radionuclide is 111 In, 99m Tc, 67 Ga, 68 Ga, 203 Pb, 64 Cu, 86 Y, 89 Zr, 123 I, 124 I, 125 I, 18 F, 76 Br, 77 Br, 152 Tb, 155 Tb, 44 Sc, and 43 A diagnostically active radionuclide selected from Sc. In certain embodiments, the radionuclide is 111 In, 99m Tc, 68 Ga, 64 Cu, 89 Zr, 123 I, 124 I, and 18 A diagnostically active radionuclide selected from F. In certain embodiments, the diagnostically active radionuclide is 68 It is Ga. In another specific embodiment, the diagnostically active radionuclide is, 18 F is F. In another specific embodiment, the diagnostically active radionuclide is, 64 It is Cu. In some embodiments, the radionuclide is 111 In, 99m Tc,94m Tc、 66 Ga、 67 Ga、 68 Ga、 52 Fe、 169 Er、 72 As、 97 Ru、 203 Pb、 61 Cu、 62 Cu、 64 Cu、 67 Cu、 89 Sr、 186 Re、 188 Re、 86 Y、 90 Y、 89 Zr、 51 Cr、 52 Mn、 51 Mn、 177 Lu、 169 Yb、 175 Yb、 105 Rh、 166 Dy、 166 Dy、 166 Ho、 153 Sm、 149 Pm、 151 Pm、 172 Tm、 121 Sn、 117 mSn、 212 Bi、 213 Bi、 142 Pr、 143 Pr、 198 Au、 199 Au、 123 I、 124 I、 125 I、 131 I、 75 Br、 76 Br、 77 Br、 80 Br、 82 Br、 18 F、 149 Tb、 152 Tb、 155 Tb、 161 Tb、 43 Sc、 44 Sc、 47 Sc、 212 Pb、 211 At、 223 Ra、 227 Th、 226 Th、82 Rb, 32 P, 76 As, 89 Zr, 111 Ag, 165 Er, 225 Ac, and 227 Selected from the group consisting of Ac. In some embodiments, the radionuclide is 111 In, 99m Tc, 67 Ga, 68 Ga, 203 Pb, 64 Cu, 86 Y, 89 Zr, 123 I, 124 I, 125 I, 18 F, 76 Br, 77 Br, 152 Tb, 155 Tb, 44 Sc, 43 Sc, 67 Cu, 188 Re, 90 Y, 177 Lu, 213 Bi, 131 I, 47 Sc, 225 Ac, 212 Pb, 211 At, or 227 Th is the radionuclide. In some embodiments, the radionuclide is 66 Ga, 67 Ga, 68 Ga, 64 Cu, 177 Lu, or 225 It is Ac. In some embodiments, the radionuclide is 111 In, 99m Tc, 68 Ga, 64 Cu, 89 Zr, 123 I, 124 I, 18 F, 90 Y, 177 Lu, 131 I, 225 Ac, 211 At, or 227 Th is the radionuclide. In certain embodiments, the radionuclide is177 It is Lu. In a particular embodiment, the radionuclide is 225 It is Ac. In certain embodiments, the radionuclide is 68 It is Ga. In certain embodiments, the radionuclide is 18 It is F.

[0255] In some embodiments, the radioactive nuclide is 177 Lu, 161 Tb, 90 Y, 67 Cu, 131 I, 225 Ac, 212 Pb, 211 At, or 227 It is Th.

[0256] In some embodiments, the radionuclide is a radiohalogen, for example, 18 F, 75 Br, 76 Br, 77 Br, 80 Br, 80m Br, 82 Br, 123 I, 124 I, 125 I, 131 I, or 211 At. When the radionuclide is a radiohalogen, the term radiohalogen includes complexes that make the radiohalogen suitable for covalent bonding to a linker or cyclic peptide, or for chelation or complex formation with a chelating agent. Such complexes assumed by the term radiohalogen include Si- 18 F, B- 18 F, and Al- 18 F is one example.

[0257] In some embodiments, the radioactive halogen is directly linked to a cyclic peptide or linker. For example, 131 I and 18 F (or any other radioactive halogen) can be substituted at any position of the linker or cyclic peptide suitable for substitution with a halo group. In some embodiments, the radioactive halogen is 18F is present. In some embodiments, when the radioactive halogen is directly linked to a cyclic peptide or linker, no chelating agent is present.

[0258] In some embodiments, the targeted constructs of this disclosure can be radiolabeled with a radionuclide at any site of the DLL3-targeting peptide. For example, in some embodiments, the DLL3-targeting peptide is directly conjugated with a radionuclide. In one embodiment, the radionuclide is covalently bonded to the DLL3-targeting peptide. In another embodiment, the radionuclide may depend on ionic interactions, thereby forming a peptide radionuclide salt.

[0259] In some embodiments, peptides targeting DLL3 can be conjugated with a chelating agent. In one embodiment, peptides targeting DLL3 can be radiolabeled, for example, by chelation of a radionuclide to a chelating agent. Chelation of a radionuclide to a chelating agent can be indicated using solid single bonds, dashed single bonds, or a combination thereof. For example, chelation of a radionuclide to DOTA can be indicated below with solid single bonds or dashed single bonds. In some embodiments, charges can also be indicated. For example, when a radionuclide is chelated with a chelating agent, each of the groups chelating the radionuclide may have a negative charge, and the radionuclide being chelated may have an opposite positive charge. Such bonds and charges are, 68 In the case of Ga, it may be shown herein as follows: [ka]

[0260] 225 In the case of AC, such bonds and charges can be exemplified (non-limiting): [ka]

[0261] In some embodiments, the peptides provided herein are radiolabeled with F-18, Ga-68, In-111, Lu-177, or Ac-225, and are pharmaceutically acceptable salts and solvates thereof. In some embodiments, the peptides are cyclic peptides. In some embodiments, the cyclic peptides are selected from compounds 94, 153, 249, and 447. In one embodiment, compound 94 is radiolabeled with F-18, Ga-68, In-111, Lu-177, or Ac-225, and is pharmaceutically acceptable salts and solvates thereof. In another embodiment, compound 153 is radiolabeled with F-18, Ga-68, In-111, Lu-177, or Ac-225, and is pharmaceutically acceptable salts and solvates thereof. In yet another embodiment, compound 249 is radiolabeled with F-18, Ga-68, In-111, Lu-177, or Ac-225, and is a pharmaceutically acceptable salt and solvate thereof. In yet another embodiment, compound 447 is radiolabeled with F-18, Ga-68, In-111, Lu-177, or Ac-225, and is a pharmaceutically acceptable salt and solvate thereof.

[0262] The radionuclides may be therapeutic radionuclides, diagnostic radionuclides, or both. Suitable radionuclides include, but are not limited to, Auger electron-emitting radionuclides, β-emitting (β-plus or β-minus emitting) radionuclides, and α-emitting (α-emitting) radionuclides. The choice of radionuclide type may depend on the use of the peptide targeting DLL3. As will be understood by those skilled in the art, several factors may be considered when selecting a radionuclide for use in a peptide targeting DLL3, such as half-life, linear energy transfer, imaging capability, and emission range in tissue. For example, β-emitting radionuclides typically have a longer emission range in tissue (e.g., 0.1–10 micrometers) and emit photons in an energy range that is easily imaged; therefore, they may be selected for use in DLL3-targeted compounds used for therapeutic, diagnostic, or theranostic purposes. On the other hand, alpha-emitting radionuclides have a shorter emission range within tissue (e.g., 50-100 micrometers) and possess high potency due to the amount of energy accumulated per unit of travel path length (i.e., linear energy transfer), which is approximately 400 times more potent than electrons (beta-minus particles) or positrons (beta-plus particles). Therefore, alpha-emitting radionuclides can be selected for therapeutic applications where the high potency of the radionuclide is desired.

[0263] Therefore, in some embodiments, the radionuclide is an α-emitting radionuclide. In other embodiments, the radionuclide is a β-emitting radionuclide. In yet another embodiment, the radionuclide is an Auger electron-emitting radionuclide.

[0264] Target structure In some embodiments, the Disclosure provides a construct that can localize to and / or associate with a target. Such a construct, including any combination of a targeting portion and a cargo, is referred to herein as a “targeting construct.” The targeting constructs provided herein may be directed towards a DLL3.

[0265] As used herein, the term “targeting moiety” refers to a component or combination of components of a targeted construct that is involved in the localization of the construct to a target or the targeting of association with a target. Cargo components of a targeted construct may include, but are not limited to, any one of a variety of compounds, including compounds, biomolecules, metals, polymer molecules, therapeutic agents, cytotoxic agents, and radioactive agents. In certain embodiments, the targeted construct comprises a targeting moiety which is a cyclic peptide targeting DLL3, which is bound to a chelating agent for the association of a radioisotope via an optional linker.

[0266] The targeted constructs of this disclosure may include chelating agents. As used herein, the term “chelating agent” refers to any compound capable of forming two or more bonds with metal atoms. Chelating agents may facilitate the association of the targeted construct with a cargo containing metal atoms. In certain embodiments, the targeted construct includes a chelating agent for the association of radioisotopes.

[0267] As used herein, the terms "chelated with" and "complexed with" mean that two independent components are linked together by one or more non-covalent bonds, such as coordination bonds.

[0268] As used herein, the terms “radiolabeled” or “labeled” mean that a non-radioactive compound is labeled with a radioactive isotope. Radiolabeling can be achieved, for example, through chelation or complexation of a chelating agent with a suitable radionuclide. Radiolabeling is, for example, 18 In the case of F, for example, it can also refer to the chemical substitution of one group on a compound with a radioactive nuclide by forming a covalent bond.

[0269] Targeted construct components may be associated via one or more linkers. For example, a targeted portion may be associated with a chelating agent or cargo via a linker. In some embodiments, the linker includes a chelating agent (e.g., if the targeted construct cargo contains metal atoms).

[0270] In some embodiments, the targeted constructs of the present disclosure include a targeted portion optionally bound by a linker to a chelating agent for the association of cargoes or cargoes. The targeted construct may comprise a single targeted portion and a single chelating agent, i.e., having the structure TM-L-CA (where "TM" is the targeted portion, "L" is an optional linker, and "CA" is a chelating agent). Alternatively, the targeted construct may comprise a construct having a single targeted portion and two or more chelating agents, for example, the structure TM-L-(CA)n (where n is an integer representing the number of chelating agents). In some embodiments, n is an integer from 1 to 50, such as 2 to 20 or 1 to 5. The targeted construct may have the structure CA-L-TM-L-CA, where each L and each CA may be the same or different.

[0271] A targeted structure optionally associated with a radioactive cargo may be referred to herein according to its corresponding analogue, i.e., a “radioactive analogue” or “non-radioactive analogue” of a given targeted structure.

[0272] In some embodiments, the targeted construct may include a detectable label. Antibody binding may be detected using the detectable label. Examples of detectable labels include, but are not limited to, radioisotopes, fluorophores, chromophores, chemiluminescent compounds, enzymes, enzyme cofactors, dyes, metal ions, ligands, biotin, avidin, streptavidin, haptens, quantum dots, or any other detectable labels known in the art or described herein.

[0273] formulation In some embodiments, the composition is administered to a human, human patient, or subject. For the purposes of this disclosure, the term “active ingredient” generally refers to the constructs described herein.

[0274] The descriptions of pharmaceutical compositions provided herein primarily concern pharmaceutical compositions suitable for administration to humans, but those skilled in the art will understand that such compositions are generally suitable for administration to any other animals, such as non-human animals, such as non-human mammals. Modifications of pharmaceutical compositions suitable for administration to humans to make them suitable for administration to various animals are well understood, and a normally skilled veterinary pharmacologist can design and / or carry out such modifications, if available, simply by ordinary experimentation. Subjects to which the administration of pharmaceutical compositions is intended include, but are not limited to, humans and / or other primates; mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, dogs, mice and / or rats; and birds, including commercially relevant birds such as poultry, chickens, ducks, geese and / or turkeys.

[0275] Formulations of the pharmaceutical compositions described herein may be prepared by any method known or to be developed in the field of pharmacology. Generally, such preparation methods include the steps of associating the active ingredient with excipients and / or one or more other accessory components, and then, if necessary and / or desirable, dividing, shaping and / or packaging the product into desired single-dose or multi-dose units.

[0276] The pharmaceutical compositions relating to this disclosure may be prepared, packaged, and / or sold together as single unit doses and / or as multiple single unit doses. As used herein, “unit dose” means an individual amount of a pharmaceutical composition containing a predetermined amount of active ingredient. The amount of active ingredient is generally equal to the dose of the active ingredient that would be administered to a subject, and / or a convenient fraction of such dose (e.g., half or one-third of such dose).

[0277] The relative amounts of the active ingredient, pharmaceutically acceptable excipients, and / or any additional ingredients in the pharmaceutical compositions according to this disclosure will vary depending on the attributes, size, and / or condition of the subject being treated, and further, on the route by which the composition is administered. For example, a composition may contain 0.1% to 100%, e.g., 0.5 to 50%, 1 to 30%, 5 to 80%, or at least 80% (w / w) of the active ingredient.

[0278] The constructs of the present disclosure may be formulated with one or more excipients to (1) increase stability; (2) enable sustained or delayed release; (3) modify biodistribution; and (4) modify the release profile of the compound in vivo. Non-limiting examples of excipients include any solvent, dispersion medium, diluent or other liquid vehicle, dispersing or suspension aid, surfactant, isotonic agent, thickener or emulsifier, and preservative. Excipients of the present disclosure may include, but are not limited to, lipidoids, liposomes, lipid nanoparticles, polymers, lipoplexes, core-shell nanoparticles, peptides, proteins, hyaluronidases, nanoparticle mimics, and combinations thereof. Accordingly, formulations of the present disclosure may contain one or more excipients in amounts that together increase the stability of the compound.

[0279] Excipients Pharmaceutical formulations may further contain pharmaceutically acceptable excipients, which, when used herein, include any solvent, dispersion medium, diluent or other liquid vehicle, dispersing or suspension aid, surfactant, isotonic agent, thickener or emulsifier, preservative, solid binder, lubricant, etc., as suitable for the specific dosage form of the desired application. Remington's *The Science and Practice of Pharmacy*, 21st Edition, ARGennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2006 (incorporated herein by reference as a whole)) discloses various excipients used in the formulation of pharmaceutical compositions and known techniques for their preparation. Unless any conventional excipient medium is incompatible with a substance or its derivative, for example, by producing some undesirable biological effect or otherwise interacting with any other component of the pharmaceutical composition in an adverse manner, its use is considered to be within the scope of this disclosure.

[0280] In some embodiments, pharmaceutically acceptable excipients are at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% pure. In some embodiments, the excipients are approved for human and veterinary use. In some embodiments, the excipients are approved by the United States Food and Drug Administration. In some embodiments, the excipients are pharmaceutical grade. In some embodiments, the excipients meet the standards of the United States Pharmacopeia (USP), European Pharmacopoeia (EP), British Pharmacopoeia, and / or International Pharmacopoeia.

[0281] Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersants and / or granulators, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants, and / or oils. Such excipients may optionally be included in the pharmaceutical composition.

[0282] Examples of diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dried starch, corn starch, powdered sugar, and / or combinations thereof.

[0283] Examples of granulating and / or dispersing agents include, but are not limited to, potato starch, corn starch, tapioca starch, sodium starch glycolate, clay, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose, and wood products, natural sponges, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinylpyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethylcellulose, cross-linked carboxymethylcellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water-insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (VEEGUM®), sodium lauryl sulfate, quaternary ammonium compounds, or combinations thereof.

[0284] Exemplary surfactants and / or emulsifiers include, but are not limited to, natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan gum, pectin, gelatin, egg yolk, casein, lanolin, tallow, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite [aluminum silicate] and VEEGUM® [magnesium aluminum silicate]), long-chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydrochloride (Cymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate [TWEEN® 20], polyoxyethylene sorbitan [TWEEN® 60], polyoxyethylene sorbitan monooleate [TWEEN® 80], sorbitan monopalmitate [SPAN® 40], sorbitan monostearate [SPAN® 60], sorbitan tristearate [SPAN® 65], glyceryl monooleate, sorbitan monooleate [SPAN® 80]), polyoxyethylene esters (e.g., polyoxyethylene monostearate [MYRJ® 45], polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and Kolliphor® (SOLUTOL®)), sucrose fatty acid esters, polyethylene glycolExamples include fatty acid esters (e.g., CREMOPHOR®), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether [BRIJ® 30]), poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, PLUORINC® F 68, poloxamer® 188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, sodium docusate, and / or combinations thereof.

[0285] Examples of binders include, but are not limited to, starches (e.g., corn starch and starch paste); gelatin; sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, and mannitol); natural and synthetic gums (e.g., acacia, sodium alginate, Irish moss extract, panwal gum, gatti gum, isapole shell mucus, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinylpyrrolidone), aluminum magnesium silicate (Veegum®), and larch-derived arabinogalactan); alginates; polyethylene oxide; polyethylene glycol; inorganic calcium salts; silicic acid; polymethacrylate; waxes; water; alcohols, etc.; and combinations thereof.

[0286] Exemplary preservatives include, but are not limited to, antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and / or other preservatives. Exemplary antioxidants include, but are not limited to, α-tocopherol, ascorbic acid, acorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and / or sodium sulfite. Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA), citrate monohydrate, disodium edetate, dipotassium edetate, edetate, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and / or trisodium edetate. Examples of antimicrobial preservatives include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidourea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercury nitrate, propylene glycol, and / or thimerosal. Examples of antifungal preservatives include, but are not limited to, butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and / or sorbic acid. Examples of alcohol preservatives include, but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and / or phenylethyl alcohol. Examples of acidic preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and / or phytic acid.Other preservatives include, but are not limited to, tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, GLYDANT PLUS®, PHENONIP®, methylparaben, GERMALL® 115, GERMABEN® II, NEOLONE®, KATHON®, and / or EUXYL®.

[0287] Examples of buffering agents include, but are not limited to, citrate buffer, acetate buffer, phosphate buffer, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dicalcium phosphate, phosphoric acid, tricalcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dsodium phosphate, monosodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogenic substance-removed distilled water, isotonic saline, Ringer's solution, ethyl alcohol, and / or combinations thereof.

[0288] Examples of lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behanate, hydrogenated vegetable oil, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and combinations thereof.

[0289] Examples of oils that contain these ingredients include, but are not limited to, almond, apricot kernel, avocado, babassu, bergamot, blackcurrant seed, borage, cade, chamomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, palm, cod liver, coffee, corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd, grape seed, hazelnut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon, and Litsea cubeba. Examples of oils include cubeba, macadamia nuts, mallow, mango seeds, meadowfoam seeds, mink, nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, camellia, savory, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, camellia, vetiver, walnut, and wheat germ oil. Exemplary oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and / or combinations thereof.

[0290] Excipients, such as cocoa butter and suppository wax, colorants, coatings, sweeteners, flavorings, and / or fragrances, may be present in the composition at the discretion of the compounder.

[0291] Administration The constructs of the present disclosure may be administered by any route that yields therapeutically effective results. These include, but are not limited to, enteral, gastrointestinal, epidural, oral, percutaneous, epidural (peridural), intracerebral (into the cerebrum), intraventricular (into the ventricular), epidermal (application to the skin), intradermal (into the skin itself), subcutaneous (into the subcutaneous), intranasal (through the nose), intravenous (into the vein), intraarterial (into the artery), intramuscular (into the muscle), intracardiac (into the heart), intramedullary injection (into the bone marrow), and intrathecal injection. These include administration (into the spinal canal), intraperitoneal, (injection or administration into the peritoneal cavity), intravesical injection, intravitreous, (through the eye), intracavernosal injection, (to the base of the penis), vaginal administration, intrauterine, extraamniotic administration, transdermal (diffusion through intact skin for systemic distribution), transmucosal (diffusion through mucous membranes), suction, sublingual, sublabial, enema, eye drops (on the conjunctiva), or ear drops. In certain embodiments, the compositions may be administered in a manner that allows them to cross the blood-brain barrier, vascular barrier, or other epithelial barriers.

[0292] The formulations described herein contain an effective amount of the construct in a pharmaceutical carrier suitable for administration to an individual requiring it. The formulations may be administered parenterally (e.g., by injection or infusion). The formulations or variations thereof may be administered in any manner via enteral, topical (e.g., into the eye), or pulmonary administration. In some embodiments, the formulations are administered topically.

[0293] dosage This disclosure provides a method comprising administering the constructs described herein to a subject in need of them. The constructs described herein may be administered to a subject in any amount and via any route of administration that is effective in preventing, treating, or imaging a disease, disorder, and / or condition (e.g., a condition associated with a disease, disorder, and / or impairment of working memory). The exact amount required will vary from subject to subject, depending on the subject's species, age, and overall condition, the severity of the disease, the specific composition, the mode of administration, and the mode of activity.

[0294] The compositions relating to this disclosure can typically be formulated in unit dosage forms for ease of administration and dose uniformity. However, it will be understood that the total daily dose of the compositions relating to this disclosure may be determined by the attending physician within the bounds of reasonable medical judgment. Specific therapeutically effective, prophylactically effective, or appropriate imaging dose levels for any particular patient will depend on a variety of factors, such as the disorder being treated and its severity; the activity of the specific compound used; the specific composition used; the patient's age, weight, overall health, sex, and diet; the time of administration, route of administration, and elimination rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors known in the medical field.

[0295] In some embodiments, the compositions according to the present disclosure are administered once or more daily in doses of approximately 0.0001 mg / kg to approximately 100 mg / kg, approximately 0.001 mg / kg to approximately 0.05 mg / kg, approximately 0.005 mg / kg to approximately 0.05 mg / kg, approximately 0.001 mg / kg to approximately 0.005 mg / kg, approximately 0.05 mg / kg to approximately 0.5 mg / kg, approximately 0.01 mg / kg to approximately 50 mg / kg, approximately 0.1 mg / kg to approximately 40 mg / kg, and approximately 0.5 mg / kg to approximately 30 mg / kg to obtain a desired therapeutic, diagnostic, preventive, or imaging effect. The drug may be administered at dose levels sufficient to deliver the target body weight / day in g / kg, approximately 0.01 mg / kg to approximately 10 mg / kg, approximately 0.1 mg / kg to approximately 10 mg / kg, or approximately 1 mg / kg to approximately 25 mg / kg, approximately 25 mg / kg to approximately 50 mg / kg, approximately 50 mg / kg to approximately 100 mg / kg, approximately 100 mg / kg to approximately 125 mg / kg, approximately 125 mg / kg to approximately 150 mg / kg, approximately 150 mg to approximately 175 mg / kg, approximately 175 mg / kg to approximately 200 mg / kg, or approximately 200 mg / kg to approximately 250 mg / kg. The desired dose may be delivered three times a day, twice a day, once a day, every other day, every three days, weekly, every two weeks, every three weeks, or every four weeks. In some embodiments, the desired dose may be delivered using multiple doses (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 doses, or more). When multiple doses are used, divided dosing regimens such as those described herein may be used.

[0296] The concentration of the construct in the pharmaceutical composition may be approximately 0.01 mg / mL to approximately 50 mg / mL, approximately 0.1 mg / mL to approximately 25 mg / mL, approximately 0.5 mg / mL to approximately 10 mg / mL, or approximately 1 mg / mL to approximately 5 mg / mL.

[0297] As used herein, “divided dose” means dividing a single unit dose or total daily dose into two or more doses, for example, two or more single unit doses. As used herein, “single unit dose” is the dose of any therapeutic agent that is administered in one dose / single dose / single route / single contact point, i.e., a single-dose event. As used herein, “total daily dose” is the amount given or prescribed over 24 hours. It may be administered as a single unit dose.

[0298] In some embodiments, for example, 177 The total dose (over the course of the treatment regimen) of a DLL3 targeted construct containing a beta radiator such as Lu is approximately 1 GBq to approximately 200 GBq. In some embodiments, the DLL3 targeted construct containing a beta radiator is administered in total dose to deliver 40 to 100 GBq of radiation. In some embodiments, the DLL3 targeted construct containing a beta radiator is administered in single doses (once every 24 hours) to deliver approximately 1 to approximately 20 GBq of radiation. In some embodiments, the DLL3 targeted construct containing a beta radiator is administered in single doses (once every 24 hours) to deliver approximately 3 to approximately 15 GBq of radiation. In some embodiments, the DLL3 targeted construct containing a beta radiator is administered in single doses (once every 24 hours) to deliver approximately 5 to approximately 10 GBq of radiation.

[0299] In some embodiments, an alpha emitter, for example, 225The total dose of a DLL3-targeted construct containing Ac (over the course of the treatment regimen) is approximately 1 MBq to approximately 100 MBq, for example, approximately 4 MBq to approximately 80 MBq, for example, approximately 5 MBq to approximately 77 MBq, for example, approximately 5 MBq, approximately 6 MBq, approximately 8 MBq, approximately 10 MBq, approximately 13 MBq, and approximately 76 MBq. In some embodiments, a DLL3-targeted construct containing an α-emitting agent is administered with a total dose of radiation of approximately 20 to approximately 80 MBq. In some embodiments, a DLL3-targeted construct containing an α-emitting agent is administered in a single dose (once within 24 hours) to deliver approximately 1 to approximately 40 MBq of radiation. In some embodiments, a DLL3-targeted construct containing an α-emitting agent is administered in a single dose (once within 24 hours) to deliver approximately 5 to approximately 40 MBq of radiation. In some embodiments, the DLL3-targeted construct containing an alpha emitter is administered in a single dose (once within 24 hours) to deliver approximately 5 to 25 MBq of radiation.

[0300] In some embodiments, an alpha emitter, for example, 225 The total dose of the DLL3-targeted construct containing Ac (over the course of the treatment regimen) is administered to the subject once every 4 to 10 weeks. In another embodiment, the construct is administered to the subject once every 6 to 8 weeks. In yet another embodiment, the construct is administered to the subject once every 6 weeks. In yet another embodiment, the construct is administered to the subject once every 6 weeks over 4 to 6 cycles.

[0301] Dosage form The pharmaceutical compositions described herein may be formulated into dosage forms described herein, such as topical, intranasal, intratracheal, or injectable (e.g., intravenous, intraocular, intravitreous, intramuscular, intracardiac, intraperitoneal, and subcutaneous).

[0302] II. Method In some embodiments, this disclosure provides methods for preparing, using, and evaluating the compounds (e.g., targeted constructs) and compositions disclosed herein.

[0303] Therapeutic indications In some embodiments, the methods of the Disclosure include methods for treating therapeutic indications using compounds and / or compositions disclosed herein. As used herein, the term “therapeutic indication” means any symptom, condition, disorder, or disease that can be alleviated, stabilized, improved, cured, or otherwise addressed by any form of treatment or other therapeutic intervention. In some embodiments, the methods of the Disclosure include treating therapeutic indications with targeted constructs disclosed herein.

[0304] In the context of disease markers or symptoms, "to reduce" or "to lessen" often means a significant decrease at a statistically significant level. The reduction could be, for example, at least 10%, at least 20%, at least 30%, or at least 40% or more, and preferably to a level that is acceptable as within the normal range for individuals without such disorder.

[0305] In the context of disease markers or symptoms, "increase" or "raise" often means a significant increase to a statistically significant level. The increase could be, for example, at least 10%, at least 20%, at least 30%, or at least 40% or more, and preferably to a level acceptable as within the normal range for individuals without such disorder.

[0306] The effectiveness of treatment or improvement of a disease can be evaluated, for example, by measuring disease progression, disease remission, symptom severity, pain reduction, quality of life, therapeutic effect, levels of disease markers, or any other measurable parameter appropriate to a given disease being treated or targeted for prevention. Monitoring the effectiveness of treatment or prevention by measuring any one of such parameters, or any combination of parameters, is well within the capabilities of those skilled in the art. In relation to the administration of the compounds or compositions described herein, "effective against" a disease or disorder means that administration in a clinically appropriate manner results in beneficial effects in at least some patients, such as improvement of symptoms, cure, reduction of disease burden, extension of life, improvement of quality of life, or other effects generally recognized positively by physicians familiar with the treatment of a particular type of disease or disorder.

[0307] Therapeutic or preventive effects are evident when there is a significant improvement, often statistically significant improvement, in one or more parameters of the disease state, or when there is no worsening or onset of symptoms that would otherwise be expected. For example, a favorable change of at least 10%, preferably at least 20%, 30%, 40%, or 50% or more, in the measurable parameters of the disease may indicate an effective treatment. The efficacy of a given compound or composition may also be determined using experimental animal models of a given disease known in the art. When using experimental animal models, the efficacy of the treatment is demonstrated when a statistically significant adjustment in markers or symptoms is observed.

[0308] In some embodiments, the methods of the present disclosure include administering the targeted constructs described herein to treat hyperproliferative disorders, metabolic disorders, infections and / or cancers. Targeted construct formulations may be administered by multiple routes, including, but not limited to, injection, oral administration or topical administration. In some embodiments, administration may be mucosal surface (lung, nose, oral, oral cavity, sublingual, vaginal, rectal) or eye (intraocular or transocular).

[0309] cancer In one embodiment, the foregoing provides a method for targeting DLL3 in a subject of interest, comprising administering a therapeutically effective amount of the compound disclosed herein to the subject.

[0310] In another embodiment, the foregoing provides a method for treating cancer in a subject of interest, comprising administering a therapeutically effective amount of one of the compounds disclosed herein to that subject.

[0311] In one embodiment, the cancer is a DLL3-mediated cancer. In another embodiment, the cancer is a DLL3-expressing cancer. In yet another embodiment, the cancer is small cell lung cancer, urothelial carcinoma, melanoma, or squamous cell carcinoma.

[0312] In another embodiment, the cancer is a neuroendocrine neoplasm, melanoma, or primary brain tumor. In another embodiment, the neuroendocrine tumor is selected from small cell lung cancer (SCLC), medullary thyroid carcinoma (MTC), large cell neuroendocrine carcinoma (LCNEC), pancreatic and gastrointestinal neuroendocrine carcinoma (GEP NEC), neuroendocrine prostate cancer (NEPC), small cell prostate cancer (SCPC), Merkel cell carcinoma (MCC), neuroendocrine cervical cancer, and grade 3 neuroendocrine tumors (NETs). In some embodiments, it is extrapulmonary neuroendocrine carcinoma (NEC) of the cervix. In some embodiments, small cell lung cancer may be extensive-stage (ES)-SCLC or limited-stage (LS)-SCLC. In some embodiments, neuroendocrine prostate cancer (NEPC) may be NEPC that has developed due to treatment.

[0313] In some embodiments, cancer may be a solid tumor that can benefit from DLL3-targeted therapy, accompanied by local demonstration of DLL3 positivity by immunohistochemistry (IHC) (e.g., ≥1% DLL3-positive cells).

[0314] In another embodiment, neuroendocrine tumors include pancreatic and gastrointestinal neuroendocrine tumors, carcinoid tumors, pheochromocytoma, paraganglioma, medullary thyroid carcinoma (MTC), pulmonary neuroendocrine tumors, thymic neuroendocrine tumors, carcinoid tumors or pancreatic neuroendocrine tumors, pituitary adenoma, adrenal tumors, Merkel cell carcinoma (MCC), breast cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, head and neck tumors, urothelial carcinoma (bladder), renal cell carcinoma, hepatocellular carcinoma, GIST, neuroblastoma, bile duct tumors, The group is selected from the following: cervical tumors, Ewing's sarcoma, osteosarcoma, small cell lung cancer (SCLC), prostate cancer, melanoma, meningioma, glioma, medulloblastoma, hemangioblastoma, supratentorial cerebellar primordial neuroectodermal tumor, olfactory neuroblastoma, functional carcinoid tumor, insulinoma, gastrinoma, vasoactive intestinal peptide (VIP)oma, glucagonoma, serotoninoma, histaminoma, ACTH-producing tumors, pheochromocytoma, and somatostatinoma.

[0315] In yet another embodiment, cancer includes acoustic neuroma, acute leukemia, acute lymphoblastic leukemia, acute myeloid leukemia (monocytic, myeloblastic, adenocarcinoma, angiosarcoma, astrocytoma, myelomonocytic and promyelocytic), acute T-cell leukemia, basal cell carcinoma, cholangiocarcinoma, bladder cancer, brain tumor, breast cancer, bronchogenic carcinoma, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (Granulocytic) leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, diffuse large B-cell lymphoma, Burkitt lymphoma, proliferative abnormalities (dysplasia and metaplasia), embryonic carcinoma, endometrial cancer, endosarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal cancer, estrogen receptor-positive breast cancer, essential thrombocythemia, Ewing's tumor, fibrosarcoma, follicular lymphoma, germ cell Testicular cancer, glioma, heavy chain disease, hemangioblastoma, liver cancer, hepatocellular carcinoma, hormone-insensitive prostate cancer, leiomyosarcoma, liposarcoma, lung cancer, intralymphatic sarcoma, lymphangiosarcoma, lymphoblastic leukemia, lymphoma (Hodgkin and non-Hodgkin), malignancies and hyperproliferative disorders of the bladder, breast, colon, lung, ovary, pancreas, prostate, skin, and uterus, lymphoid malignancies of T-cell or B-cell origin, leukemia, lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloid leukemia, myeloma, The group is selected from myxosarcoma, neuroblastoma, non-small cell lung cancer, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pineal glandoma, polycythemia vera, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous carcinoma, seminomas, skin cancer, small cell lung cancer, solid tumors (carcinomas and sarcomas), small cell lung cancer, gastric cancer, squamous cell carcinoma, synoviomas, sweat gland carcinoma, thyroid cancer, Waldenström macroglobulinemia, testicular tumors, uterine cancer, and Wilms' tumor.

[0316] In another embodiment, cancer includes primary cancer, metastatic cancer, oropharyngeal cancer, hypopharyngeal cancer, liver cancer, gallbladder cancer, bile duct cancer, small intestine cancer, urinary tract cancer, kidney cancer, urothelial carcinoma, female reproductive organ cancer, uterine cancer, gestational trophoblastic disease, male reproductive organ cancer, seminal vesicle cancer, testicular cancer, germ cell tumor, endocrine gland tumor, thyroid cancer, adrenal gland cancer, pituitary cancer, hemangioma, sarcoma originating from bone and soft tissue, Kaposi's sarcoma, nerve cancer, eye cancer, meningeal cancer, gliablastoma, neuroma, neuroblastoma, schwannoma, solid tumors originating from hematopoietic malignancies, such as leukemia, metastatic melanoma, recurrent or persistent epithelial ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gastrointestinal stromal tumor, colorectal cancer, gastric cancer, melanoma, glioblastoma multiforme, non-squamous non-small cell lung cancer, malignant glioma, The following are selected from the group consisting of epithelial ovarian cancer, primary peritoneal serous carcinoma, metastatic liver cancer, neuroendocrine carcinoma, refractory malignant tumors, triple-negative breast cancer, HER2-amplified breast cancer, nasopharyngeal carcinoma, oral cancer, biliary tract cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma (SCCHN), non-medullary thyroid carcinoma, recurrent glioblastoma multiforme, neurofibromatosis type 1, CNS cancer, liposarcoma, leiomyosarcoma, salivary gland cancer, mucosal melanoma, acral / lentiginous melanoma, paraganglioma, pheochromocytoma, advanced metastatic cancer, solid tumors, triple-negative breast cancer, colorectal cancer, sarcoma, melanoma, renal cancer, endometrial cancer, thyroid cancer, rhabdomyosarcoma, multiple myeloma, ovarian cancer, glioblastoma, gastrointestinal stromal tumors, mantle cell lymphoma, and refractory malignant tumors.

[0317] In one embodiment, cancer is selected from the group consisting of breast, ovary, cervix, prostate, testis, genitourinary tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanocyte, lung, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung adenocarcinoma, bone, colon, colorectal, adenoma, pancreas, adenocarcinoma, thyroid, follicular adenocarcinoma, anaplastic carcinoma, papillary carcinoma, seminomas, melanoma, sarcoma, bladder cancer, liver and biliary tract cancer, kidney cancer, myeloid disorders, lymphoid disorders, Hodgkin's disease, pilocytic cells, oral cavity and pharynx (oral cavity), lips, tongue, mouth, pharynx, small intestine, colon, rectum, large intestine, rectum, brain and central nervous system, chronic myeloid leukemia (CML), and leukemia.

[0318] In another embodiment, the cancer is selected from the group consisting of myeloma, lymphoma, or cancers selected from gastric cancer, renal cancer, head and neck cancer, oropharyngeal cancer, non-small cell lung cancer (NSCLC), endometrial cancer, liver cancer, non-Hodgkin lymphoma, and lung cancer.

[0319] In one embodiment, the cancer is selected from the group consisting of prostate cancer, colon cancer, lung cancer, head and neck squamous cell carcinoma, esophageal cancer, hepatocellular carcinoma, melanoma, sarcoma, gastric cancer, pancreatic cancer, ovarian cancer, and breast cancer.

[0320] In one embodiment, cancer is selected from the group consisting of tumors, neoplasms, carcinomas, sarcomas, leukemias, lymphomas, etc. For example, cancers include, but are not limited to, mesothelioma, leukemia and lymphoma, such as cutaneous T-cell lymphoma (CTCL), non-cutaneous peripheral T-cell lymphoma, lymphoma associated with human T-cell lymphotropic virus (HTLV), such as adult T-cell leukemia / lymphoma (ATLL), B-cell lymphoma, acute non-lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, lymphoma, and multiple myeloma, non-Hodgkin lymphoma, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), Hodgkin lymphoma, Burkitt lymphoma, adult T-cell leukemia / lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), or hepatocellular carcinoma. Further examples include myelodysplastic syndromes, pediatric solid tumors such as brain tumors, neuroblastomas, retinoblastomas, Wilms' tumors, bone tumors, and soft tissue sarcomas, common solid tumors in adults such as head and neck cancers (e.g., oral cancer, laryngeal cancer, nasopharyngeal cancer, and esophageal cancer), genitourinary cancers (e.g., prostate cancer, bladder cancer, kidney cancer, uterine cancer, ovarian cancer, testicular cancer), lung cancers (e.g., small cell and non-small cell), breast cancer, pancreatic cancer, melanoma and other skin cancers, gastric cancer, brain tumors, tumors associated with Gorin syndrome (e.g., medulloblastoma, meningioma, etc.), and liver cancer. Further exemplary forms of cancer that can be treated with the compounds of the present invention include, but are not limited to, cancers of the skeletal or smooth muscle, gastric cancer, small intestine cancer, rectal cancer, salivary gland cancer, endometrial cancer, adrenal cancer, anal cancer, rectal cancer, parathyroid cancer, and pituitary cancer.

[0321] Further cancers for which the compounds described herein may be useful for treatment include, for example, colorectal cancer, familial adenomatous polyposis cancer and hereditary nonpolyposis colorectal cancer, or melanoma. Furthermore, cancers that may be useful include, but are not limited to, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, thyroid cancer (medullary and papillary thyroid carcinoma), kidney cancer, renal parenchymal cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, testicular cancer, urinary tract cancer, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermal tumors, gallbladder cancer, bronchial cancer, multiple myeloma, basal cell tumor, teratoma, retinoblastoma, choroidal melanoma, seminomas, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myasthenia, liposarcoma, fibrosarcoma, Ewing's sarcoma, and plasmacytoma.

[0322] In another aspect, the disclosure provides compounds disclosed herein, or pharmaceutically acceptable salts thereof, for use in the manufacture of agents for treating diseases in which DLL3 plays a role.

[0323] One aspect of this disclosure provides compounds useful for treating diseases, disorders, and conditions characterized by excessive or abnormal cell proliferation. Such diseases include, but are not limited to, proliferative or hyperproliferative diseases and neurodegenerative diseases. Examples of proliferative and hyperproliferative diseases include, but are not limited to, cancer.

[0324] In another aspect, what is provided herein is the use of one or more compounds of this disclosure in the manufacture of agents for the treatment of cancer, including, but not limited to, various types of cancer disclosed herein.

[0325] In some embodiments, therapeutic indications include cancer-related indications. The term “cancer” refers to a group of diseases characterized by dysfunctional cell proliferation and division, and possibly spreading between regions of the body. As used herein, the term “cancer-related indication” refers to any disease, disorder, or condition relating to cancer, cancer treatment, or precancerous conditions. Cancer-related indications include, but are not limited to, pathological conditions characterized by malignant neoplastic growth, tumors, and / or hematological malignancies. In some embodiments, the methods of the Disclosure include the treatment of cancer-related indications with the targeted constructs of the Disclosure.

[0326] In various embodiments, methods are provided for treating cancer, comprising administering a therapeutically effective amount of a construct described herein, or a salt form thereof, to a subject having cancer, suspected of having cancer, or predisposed to cancer. According to this disclosure, cancer encompasses any disease or illness characterized by uncontrolled cell proliferation, such as hyperproliferation. Cancer may be characterized by a tumor, such as a solid tumor, or any neoplasm.

[0327] In some embodiments, the subject may not have indications for treatment with the construct. In some embodiments, the method involves the use of cancer cells, including, but not limited to, mammalian cancer cells. In some examples, the mammalian cancer cells are human cancer cells.

[0328] In some embodiments, the constructs according to this disclosure inhibit cancer and / or tumor growth. They also reduce one or more of cell proliferation, invasiveness, and metastasis, thereby being useful in cancer treatment.

[0329] In some embodiments, the constructs described herein may be used to prevent the growth of tumors or cancers and / or to prevent the metastasis of tumors or cancers. In some embodiments, the compositions described herein may be used to shrink or destroy cancers.

[0330] In some embodiments, the constructs provided herein are useful for inhibiting the proliferation of cancer cells. In some embodiments, the constructs provided herein are useful for inhibiting cell proliferation, e.g., inhibiting the rate of cell proliferation, preventing cell proliferation, and / or inducing cell death. Generally, the constructs described herein can inhibit the proliferation of cancer cells, or can both inhibit the proliferation of cancer cells and / or induce cell death. In some embodiments, cell proliferation is reduced by at least about 25%, about 50%, about 75%, or about 90% after treatment with the constructs of the disclosure compared to untreated cells. In some embodiments, the cell cycle arrest marker phosphorylated histone H3 (PH3 or PHH3) is increased by at least about 50%, about 75%, about 100%, about 200%, about 400%, or about 600% after treatment with the constructs of the disclosure compared to untreated cells. In some embodiments, cell apoptosis marker cleavage caspase 3 (CC3) increases by at least 50%, about 75%, about 100%, about 200%, about 400%, or about 600% after treatment with the constructs of the present disclosure compared to untreated cells.

[0331] Furthermore, in some embodiments, the constructs of the present disclosure are effective in inhibiting tumor growth in multiple types of tumors, whether measured as a net size (weight, surface area, or volume) or as a rate over time.

[0332] In some embodiments, the size of the tumor is reduced by at least about 60% after treatment with the construct of the present disclosure. In some embodiments, the size of the tumor is reduced by at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, and at least about 100%, in terms of weight and / or area and / or volume.

[0333] Cancers treatable by the methods described herein generally occur in mammals. Examples of mammals include humans, non-human primates, dogs, cats, rats, mice, rabbits, ferrets, guinea pigs, horses, pigs, sheep, goats, and cattle. In various embodiments, cancers may include, but are not limited to, acoustic neuroma, acute leukemia, acute lymphoblastic leukemia, acute myeloid leukemia (monocytic, myeloblastic, adenocarcinoma, angiosarcoma, astrocytoma, myelomonocytic and promyelocytic), acute T-cell leukemia, basal cell carcinoma, cholangiocarcinoma, bladder cancer, brain tumors, breast cancer, bronchogenic carcinoma, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, and chronic Myeloid leukemia (granulocytic leukemia), chronic myeloid leukemia, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, diffuse large B-cell lymphoma, Burkitt lymphoma, proliferative abnormalities (dysplasia and metaplasia), embryonic carcinoma, endometrial cancer, endometrial sarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal cancer, estrogen receptor-positive breast cancer, essential thrombocythemia, Ewing's tumor, fibrosarcoma, follicular lymphoma, germ cell Testicular cancer, glioma, heavy chain disease, hemangioblastoma, liver cancer, hepatocellular carcinoma, hormone-insensitive prostate cancer, leiomyosarcoma, liposarcoma, lung cancer, intralymphatic sarcoma, lymphangiosarcoma, lymphoblastic leukemia, lymphoma (Hodgkin and non-Hodgkin), malignancies and hyperproliferative disorders of the bladder, breast, colon, lung, and ovaries, lymphoid malignancies of T-cell or B-cell origin of the pancreas, prostate, skin, and uterus, leukemia, lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloid leukemia, bone These include myeloma, myxosarcoma, neuroblastoma, non-small cell lung cancer, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pineal glandoma, polycythemia vera, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous carcinoma, seminomas, skin cancer, small cell lung cancer, solid tumors (carcinomas and sarcomas), small cell lung cancer, gastric cancer, squamous cell carcinoma, synoviomas, sweat gland carcinomas, thyroid cancer, Waldenström macroglobulinemia, testicular tumors, uterine cancer, and Wilms' tumor.Other cancers include primary cancer, metastatic cancer, oropharyngeal cancer, hypopharyngeal cancer, liver cancer, gallbladder cancer, bile duct cancer, small intestine cancer, urinary tract cancer, kidney cancer, urothelial carcinoma, female reproductive organ cancer, uterine cancer, gestational trophoblastic disease, male reproductive organ cancer, seminal vesicle cancer, testicular cancer, germ cell tumors, endocrine gland tumors, thyroid cancer, adrenal cancer, pituitary cancer, hemangioma, sarcomas of bone and soft tissue origin, Kaposi's sarcoma, nerve cancer, eye cancer, meningeal cancer, gliablastoma, neuroma, neuroblastoma, schwannoma, solid tumors of hematopoietic malignancies, such as leukemia, metastatic melanoma, recurrent or persistent epithelial ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gastrointestinal stromal tumors, colorectal cancer, gastric cancer, melanoma, glioblastoma multiforme, non-squamous non-small cell lung cancer, and malignant glioma. These include epithelial ovarian cancer, primary serous peritoneal carcinoma, metastatic liver cancer, neuroendocrine carcinoma, refractory malignancies, triple-negative breast cancer, HER2-amplified breast cancer, nasopharyngeal cancer, oral cancer, biliary tract cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma (SCCHN), non-medullary thyroid carcinoma, recurrent glioblastoma multiforme, neurofibromatosis type 1, CNS cancer, liposarcoma, leiomyosarcoma, salivary gland cancer, mucosal melanoma, acral / lentiginous melanoma, paraganglioma, pheochromocytoma, advanced metastatic cancer, solid tumors, triple-negative breast cancer, colorectal cancer, sarcoma, melanoma, renal cancer, endometrial cancer, thyroid cancer, rhabdomyosarcoma, multiple myeloma, ovarian cancer, glioblastoma, gastrointestinal stromal tumors, mantle cell lymphoma, and refractory malignancies.

[0334] In some embodiments, the targeted constructs of the Disclosure are used to target cancer cells expressing DLL3. In some embodiments, the targeted constructs of the Disclosure are used to treat lung cancer, breast cancer, bladder cancer, colon cancer, urothelial carcinoma, melanoma, or squamous cell carcinoma.

[0335] Theranostics The term "therapeutic," derived from the combination of therapeutic and diagnostic agents, represents a new field in medicine where specific disease-targeting agents, such as radiopharmaceuticals, can be used to simultaneously or sequentially diagnose and treat medical conditions. Theranostics has become an important area of ​​research and development in medical physics, where a given disease-targeting agent, such as a radioligand therapy, can be transformed from an imaging probe (e.g., by using β+ or γ-emitting isotopes to facilitate positron emission tomography (PET) or single-photon emission computed tomography (CT) imaging, respectively) to a therapeutic probe (e.g., by using α or β particles or Auger electron-emitting isotopes to facilitate targeted radiotherapy) by altering the isotopes of the radionuclides present in the agent.

[0336] Molecular imaging is a well-known and useful technique for in vivo diagnostics. It can be used in a wide variety of ways, including three-dimensional mapping of molecular processes such as gene expression, blood flow, physiological changes (e.g., pH), immune responses, and cell transport. It can be used to detect and diagnose diseases, select optimal treatments, monitor treatment effectiveness, and obtain early reads of efficacy.

[0337] In principle, several different techniques, including PET, single-photon emission tomography (SPET), magneto-optical resonance imaging (MRI), CT, and Cherenkov emission imaging (CLI), can be used for molecular imaging. Combinations of these modalities are emerging to provide improved clinical applications, such as PET / CT and SPET / CT ("multimodal imaging").

[0338] Radionuclide imaging using PET and SPET offers the advantages of extremely high sensitivity and the administration of small amounts of contrast agent (e.g., picomolar concentration in vivo), which does not disrupt molecular processes in vivo. Furthermore, the targeting principle for radionuclide imaging can also be applied to the targeted delivery of radionuclide therapy. Typically, isotopes used as radionuclides in molecular imaging or therapy are incorporated into molecules to generate pharmaceutically acceptable radiotracers for the target.

[0339] Therefore, the structures of this disclosure can be used in radiotherapy and diagnostic medical imaging.

[0340] Combination therapy In some embodiments, the constructs of the present disclosure are combined with at least one additional activator. The activator may be any suitable drug. The activator may be selected from the group consisting of hormonal agents, anti-cancer agents, chemotherapeutic agents, immunotherapeutic agents, immunomodulators, radiosensitizers, DNA damage repair inhibitors, PARP (poly-ADP-ribose polymerase) inhibitors, and combinations thereof. The construct and at least one additional activator may be administered simultaneously, sequentially, or in any order. The construct and at least one additional activator may be administered in different doses, at different frequencies, or via different routes, whichever is appropriate.

[0341] In some embodiments, additional activators may affect the biodistribution (i.e., tissue distribution) of the constructs of the Disclosure. For example, radioactive agents may accumulate in the kidneys and cause potential radiotoxicity problems in the kidneys and surrounding organs. Additional activators may reduce renal accumulation or residence time. Preferably, renal uptake of the construct is reduced, while tumor uptake of the construct remains unaffected. The kidneys and surrounding organs are protected without reducing the efficacy of the construct. In one non-limiting example, the constructs of the Disclosure may be administered in combination with at least one amino acid or its analogue. The amino acid or its analogue may be a positively charged basic amino acid such as lysine (L-lysine or D-lysine) or arginine, or a combination thereof.

[0342] The additional activator may also be selected from any activators described herein, such as drugs for treating cancer. It may also be a cancer symptom reliever. Non-limiting examples of symptom relievers include octreotide or lanreotide; interferon, cypoheptadine, or any other antihistamine. In some embodiments, the constructs of the Disclosure do not have drug-drug interference with the additional activator. The additional activator may be administered concurrently with the constructs of the Disclosure.

[0343] In some embodiments, non-radioactive analogs of the constructs of the Disclosure may be combined with radioactive analogs of the constructs. For example, the non-radioactive construct may be administered before the radioactive analog. In another example, a subject may be given a mixture of the non-radioactive construct and its radioactive analog. In yet another example, a subject may be given a non-radioactive construct treatment first, followed by a mixture of the non-radioactive construct and its radioactive analog.

[0344] In some embodiments, a construct of the Disclosure comprising one radioactive label may be combined with at least one other construct of the Disclosure comprising one or more different radioactive labels. For example, a construct comprising an imaging radioactive label may be combined with a construct comprising a non-imaging radioactive label. In one embodiment, a construct associated with lutetium (Lu) may be combined with a construct associated with gallium (Ga).

[0345] The constructs described herein or formulations containing the constructs described herein may be used for the selective tissue delivery of therapeutic, prophylactic, or diagnostic agents to individuals or patients in need. For example, the constructs of the Disclosure may be used to deliver a radioagent to selected tissue, which may be tumor tissue. The administration regimen may be adjusted to produce the desired optimal response (e.g., a therapeutic or prophylactic response). For example, a single bolus may be administered, or divided doses may be administered over a period of time, or the dose may be proportionally reduced or increased as indicated by the requirements of the treatment situation. As used herein, unit dosage form refers to a physically distinct unit suitable as a unit dose for the mammalian subject to be treated; each unit contains a predetermined amount of the active compound calculated to produce the desired therapeutic effect.

[0346] Diagnostic use In some embodiments, the Disclosure provides diagnostic methods comprising the use of targeted moieties and / or targeted constructs. Such methods may include detecting DLL3 using any of the targeted moieties and / or targeted constructs described herein. Such methods may include contacting a subject or subject sample with a targeted moiety and / or targeted construct described herein. The peptides and / or targeted constructs may bind to DLL3. The targeted moieties and / or targeted constructs used in the detection method may include detectable labels. The detection method may include the use of detection reagents for detecting the bound antibody or peptide. As used herein, the term “detection reagent” means any compound or substance used to visualize or observe a subject (e.g., a bound antibody or a detectable label) or event. The detection reagent may include a secondary antibody or other high-affinity compound (e.g., biotin or avidin) that binds to the antibody or associated conjugate to be detected. The detection reagent may be or include a substrate for the detection of an enzymatically detectable label (e.g., associated with a primary or secondary antibody).

[0347] The diagnostic applications of this disclosure may include detecting DLL3 in a sample of interest containing cells. In some embodiments, cell-associated DLL3 may be detected. Cell-associated DLL3 may be detected in the sample of interest by fluorescence-associated cell sorting (FACS) analysis. In some embodiments, DLL3 may be detected in the sample of interest by immunohistochemistry. Such methods may include the use of a colorimetric analysis-based system or an immunofluorescence-based system for DLL3 detection.

[0348] In some embodiments, the Disclosure provides a method for stratifying subjects based on the detection of DLL3 in a subject or subject sample. Such a method may include detecting DLL3 in a subject or subject sample using one of the methods described herein (for example, using a peptide or a peptide-containing targeted construct) and classifying the subjects according to the detected level of DLL3. In certain embodiments, the targeted construct includes a targeted moiety which is a cyclic peptide that targets DLL3.

[0349] In some embodiments, subjects may be classified according to the presence or absence of DLL3 in the subject or subject sample and / or the level of DLL3. Subjects may be further classified according to the presence or absence of specific DLL3 extracellular subdomains in the subject or subject sample and / or the level of specific DLL3 extracellular subdomains. Classifications used in stratifying subjects may include, but are not limited to, disease type, disease prognosis or severity, suitability of treatment, and the type of treatment most likely to be successful or appropriate.

[0350] III. Kits and Devices This disclosure provides various kits and devices for conveniently and / or effectively carrying out the methods of this disclosure. Typically, the kits include a sufficient quantity and / or number of components to enable a user to perform multiple treatments of the subject and / or to perform multiple experiments.

[0351] In one embodiment, the Disclosure provides a kit for inhibiting cancer cell proliferation in vitro or in vivo, comprising a construct of the Disclosure or a combination of the constructs of the Disclosure in any combination with any other activator.

[0352] The kit may further include packaging and instructions for forming the formulation composition and / or a delivery agent. The delivery agent may include saline, a buffer, or any delivery agent disclosed herein. The amounts of each component may be varied to enable consistent and reproducible higher concentrations of saline or simple buffer formulations. The components may also be varied to increase the stability of the construct in the buffer over a period of time and / or under various conditions.

[0353] This disclosure provides devices that may incorporate constructs of the disclosure. These devices contain stable formulations that are ready for immediate delivery to subjects requiring them, such as human patients. In some embodiments, the subjects have cancer.

[0354] Non-limiting examples of devices include pumps, catheters, needles, transdermal patches, pressurized olfactory organ delivery devices, iontophoresis devices, and multilayer microfluidic devices. Devices may be used to deliver the constructs of the Disclosure according to single, multiple, or divided dosing regimens. Devices may be used to deliver the constructs of the Disclosure intradermally, subcutaneously, or intramuscularly throughout biological tissue.

[0355] IV. Definition The following are definitions of various terms used to describe the compounds and compositions disclosed herein. These definitions apply to the terms as they are used herein and in the claims, individually or as part of a larger group, unless otherwise specified.

[0356] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art. Generally, the nomenclature used herein and the experimental procedures in cell culture, molecular genetics, organic chemistry, and peptide chemistry described herein are well known and commonly used in the art.

[0357] As used herein, articles ("a" and "an") refer to one or more (i.e., at least one) grammatical objects of the article. For example, "an element" means one or more elements. Furthermore, the use of the term "including" and other forms such as "include," "includes," and "included" is not limited.

[0358] As used herein, the term “about” will be understood by those skilled in the art and will vary to some extent depending on the context in which it is used. As used herein, when referring to a measurable value such as a quantity or length of time, the term “about” means to include a variation of ±20% or ±10% (including ±5%, ±1%, and ±0.1%) from a specified value, provided that the variation is appropriate for carrying out the method of this disclosure.

[0359] As used herein, the term "administration," etc., refers to providing a therapeutic agent to a subject. In the art, there are many methods of administering therapeutic agents, including, but are not limited to, intravenous administration, oral administration, aerosol administration, parenteral administration, intraocular administration, intrapulmonary administration, and topical administration.

[0360] The term "alkylene," used alone or in combination with other terms, refers to a divalent alkyl linking group. Formally, an alkylene group corresponds to an alkane in which two CH bonds are substituted by bonding sites to the remainder of the alkylene compound. n~m "Alkylene" refers to an alkylene group having n to m carbon atoms. Examples of alkylene groups, though not limited to them, include methylene, ethane-1,2-diyl, ethane-1,1-diyl, propane-1,3-diyl, propane-1,2-diyl, propane-1,1-diyl, butane-1,4-diyl, butane-1,3-diyl, butane-1,2-diyl, and 2-methyl-propane-1,3-diyl.

[0361] As used herein, the terms “associated with,” “conjugated,” “linked,” “bonded,” and “tethered” mean, when used in reference to two or more entities, that they are physically associated or linked to one or more entities, either directly or via one or more parts acting as linkers, to form a structure that is sufficiently stable for the entities to remain physically associated, for example, under operating conditions, for example, under physiological conditions. “Association” does not have to be by covalent chemical bonds and may include other forms of association or bonding that are sufficiently stable for the “associated” entities to remain physically associated, such as ionic or hydrogen bonding or hybridization-based bonding.

[0362] In some embodiments, the Disclosure provides a method for stratifying subjects based on the detection of DLL3 in a subject or subject sample. Such a method may include detecting DLL3 in a subject or subject sample using one of the methods described herein (for example, using a peptide or a peptide-containing targeted construct) and classifying the subjects according to the detected level of DLL3. In certain embodiments, the targeted construct includes a targeted moiety which is a cyclic peptide that targets DLL3.

[0363] As used herein, the term "cancer" refers to a disease characterized by abnormal cell proliferation and division.

[0364] As used herein, the term "cancer cell" refers to a cell that proliferates and divides in an abnormal and uncontrolled manner.

[0365] As used herein, the term “compound” refers to a distinct chemical entity. Constructs, targeted constructs, targeted portions, cargoes, chelating agents, or other construct components may be referred to as compounds, individually or collectively, together with any of the aforementioned fragments or variants.

[0366] Compounds may exist in one or more isomeric or isotopic forms (including, but not limited to, stereoisomers, geometric isomers, tautomers, and isotopes). Compounds may be provided or used in singular form or as mixtures of two or more forms (including, but not limited to, racemic mixtures of stereoisomers). Some compounds may exist in different forms, and they may exhibit different properties and / or activities (including, but not limited to, biological activity). For example, compounds containing asymmetrically substituted carbon atoms may be isolated in optically active or racemic forms. As used herein, the following structures indicate the presence of a double bond, and the substituents may be configured as E or Z isomers: [ka]

[0367] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise specified, all stereoisomers, including enantiomers and diastereomers, are intended. Compounds of this disclosure containing asymmetrically substituted carbon atoms may be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically active starting materials, such as by decomposition of racemic mixtures or stereoselective synthesis, are known in the art. Many geometric isomers of olefins, C=N double bonds, etc., may also be present in the compounds described herein, and all such stable isomers are conceivable in this disclosure. The cis and trans geometric isomers of the compounds of this disclosure may be isolated as mixtures of isomers or as separated isomers.

[0368] Tautomer compound forms arise from the exchange of a single bond with an adjacent double bond and the resulting transfer of protons. Tautomers include prototropic tautomers, which are isomeric protonated states having the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imoid acid pairs, lactam-lactim pairs, amide-imoid acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions in the heterocyclic system, such as 1H- and 3H-imidazoles, 1H-, 2H- and 4H-1,2,4-triazoles, 1H- and 2H-isoindole, and 1H- and 2H-pyrazoles. Tautomers may exist in equilibrium or be sterically fixed into one form by appropriate substitution.

[0369] The compounds described herein may be provided in forms containing different isotopes of the compound atoms. “Isotopes” refer to atoms that have the same atomic number but different mass numbers due to different numbers of neutrons in their nuclei. For example, isotopes of hydrogen include tritium and deuterium.

[0370] The compounds described herein may be provided as salts, or may be prepared by conventional methods in combination with a solvent or water molecule to form solvates and hydrates.

[0371] As used herein, the term “hydrate” refers to a complex formed by combining a compound of formula I, formula C, or any of the formulas disclosed herein with water.

[0372] The term "solvate" refers to a complex formed by combining a compound of formula I, formula C, or any other formula disclosed herein with a solvent or a crystalline solid containing an amount of solvent incorporated into its crystalline structure. As used herein, the term "solvate" includes hydrates.

[0373] As used herein, the term “construction” refers to an artificially manipulated molecule. Some constructs may include nucleic acids and / or peptides, which may be products of recombinant technology and may be artificially synthesized or expressed from recombinant nucleic acid sequences. Constructs may be combinations of nucleic acids, peptides, and / or other compounds.

[0374] As used herein, the term “cyclic” refers to the presence of a continuous loop. A cyclic molecule does not have to be cyclic; it is simply linked together to form an unbroken chain of subunits. A cyclic peptide may contain a “cyclic loop” formed when two amino acids are linked by a crosslinking site. The cyclic loop contains amino acids along the peptide present between the crosslinking amino acids. A cyclic loop may contain 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acids.

[0375] As used herein, the terms “effective dose,” “pharmaceutical effective dose,” and “therapeutic effective dose” refer to an amount of a drug that is nontoxic but sufficient to produce a desired biological effect. This effect may be a reduction or alleviation of signs, symptoms, or causes of a disease, or any other desired change in the biological system. In any individual case, the appropriate therapeutic dose can be determined by those skilled in the art using routine experiments.

[0376] As used herein, “epitope” refers to a surface or region on one or more entities that can interact with an antibody or other bound biomolecule. For example, a protein epitope may contain one or more amino acids and / or post-translational modifications (e.g., phosphorylated residues) that interact with an antibody. In some embodiments, an epitope may be a “conformational epitope,” which refers to an epitope that includes a specific three-dimensional configuration of an entity that has or forms an epitope. For example, a protein conformational epitope may include a combination of amino acids and / or post-translational modifications from folded nonlinear stretches of amino acid chains.

[0377] As used herein, the terms “equilibrium dissociation constant” or “K” are used. D" refers to a value representing the tendency of two or more agents (e.g., two proteins) to reversibly separate. In certain cases, K D indicates the concentration of the first agent at which half of the total level of the second agent binds to the first agent.

[0378] As used herein, "expression" of a nucleic acid sequence refers to one or more of the following events: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of the RNA transcript (e.g., by splicing, editing, 5' cap formation, and / or 3' end processing); (3) translation of the RNA into a peptide or protein; and (4) post-translational modification of the peptide or protein.

[0379] As used herein, the term "half-life" or "t 1 / 2 " refers to the time it takes for a given process or compound concentration to reach half of its final value. "Terminal half-life" or "terminal t 1 / 2 " refers to the time required for the plasma concentration of a factor to decrease by half after the plasma concentration of the factor has reached pseudo-equilibrium.

[0380] As used herein, the term "halo" or "halogen" means a fluorine, chlorine, bromine, or iodine atom, preferably fluorine, chlorine, or bromine, more preferably fluorine or chlorine, alone or as part of another substituent, unless otherwise specified.

[0381] As used herein, the term “identity” refers to the comparative relationship between sequences when referring to peptides or nucleic acids. This term is used to describe the degree of sequence relevance between polymer sequences and may include the proportion of monomer components that match a gap alignment (if any) addressed by a specific mathematical model or computer program (i.e., “algorithm”). The identity of related peptides can be readily calculated by known methods. Such methods include, but are not limited to, those previously described by other authors (Lesk, AM, ed., Computational Molecular Biology, Oxford University Press, New York, 1988; Smith, DW, ed., Biocomputing: Informatics and Genome Projects, Academic Press, New York, 1993; Griffin, AM et al., ed., Computer Analysis of Sequence Data, Part 1, Humana Press, New Jersey, 1994; von Heinje, G., Sequence Analysis in Molecular Biology, Academic Press, 1987; Gribskov, M. et al., ed., Sequence Analysis Primer, M. Stockton Press, New York, 1991; and Carillo et al., Applied Math, SIAM J, 1988, 48, 1073).

[0382] As used herein, the term "lactam crosslink" refers to an amide bond that forms a crosslink between chemical groups in a molecule. In some cases, lactam crosslinks are formed between amino acids in a peptide.

[0383] As used herein, “linker” refers to any chemical structure that links two or more entities or domains. A linker may contain one or more chemical bonds, atoms, atomic groups, and / or chemical groups. Examples of chemical groups that may be included in a linker include, but are not limited to, alkyl, alkenyl, alkynyl, amide, amino, ether, thioether, ester, alkylene, heteroalkylene, aryl, or heterocyclyl chemical groups, each of which may be optionally substituted as described herein. A linker may contain one or more of the following: unsaturated alkanes, polyethylene glycols (e.g., ethylene or propylene glycol monomer units, e.g., diethylene glycol, dipropylene glycol, triethylene glycol, tripropylene glycol, tetraethylene glycol, or tetraethylene glycol), and dextran polymers. A linker may contain amino acids, peptides, polypeptides, and / or proteins.

[0384] The linker carbon chain length may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 atoms, or more. The linker carbon chain may contain heteroatoms (e.g., nitrogen, oxygen, sulfur, etc.).

[0385] The entities or domains linked by the linker may include, but are not limited to, atoms, chemical groups, nucleosides, nucleotides, nucleic acid bases, sugars, nucleic acids, amino acids, peptides, proteins, protein complexes, cargoes, therapeutic agents, and detectable labels. The linker may be used for multiple purposes, including, but are not limited to, forming a polymer or conjugate. For example, compounds intended by this disclosure may include two or more targeting agents and / or two or more cargoes. For example, the cyclic peptides disclosed herein may include two or more chelating agents, and therefore two or more radionuclides. As another example, the constructs disclosed herein may include two or more of the targeted cyclic peptides disclosed herein.

[0386] The linker may include cleavable elements, such as disulfide (-SS-) bonds or azo (-N=N-) bonds that can be cleaved using a reducing agent or photolysis. Selectively cleavable bonds may include amide bonds that can be cleaved, for example, by photolysis or by using tris(2-carboxyethyl)phosphine (TCEP) or other reducing agents. Selectively cleavable bonds may include ester bonds that can be cleaved, for example, by acidic or basic hydrolysis.

[0387] Examples of linkers, though not limited to them, include pH-sensitive linkers, protease-cleavable peptide linkers, nuclease-sensitive nucleic acid linkers, lipase-sensitive lipid linkers, glycosidase-sensitive carbohydrate linkers, hypoxia-sensitive linkers, photocleavable linkers, thermally unstable linkers, enzyme-cleavable linkers (e.g., esterase-cleavable linkers), ultrasound-sensitive linkers, and X-ray-cleavable linkers.

[0388] As used herein, the term "peptide skeleton" consists of repeating units of an amino group, an α-carbon, and a carbonyl group (e.g., -NH2-CH-C(O-)).

[0389] As used herein, the term “modulation” refers to upregulation (i.e., activation or stimulation) or downregulation (i.e., inhibition or suppression) of a response, or two of them in combination or separately. Modulation is generally compared to a baseline or reference, which may be internal or external to the entity being treated.

[0390] As used herein, the term “patient” refers to an object seeking, needing, in need of, receiving, expecting treatment, or under the care of a trained (e.g., licensed) professional for a particular disease, disorder, or condition. A patient may include any living organism. Treatment of a patient may include, but is not limited to, experimental, diagnostic, prophylactic, and / or therapeutic treatments. Typical patients may include, but is not limited to, animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans).

[0391] As used herein, the term “pharmaceutical composition” means a composition comprising at least one active ingredient in a form and amount that enables the active ingredient to be therapeutically effective. Pharmaceutical compositions facilitate the administration of a compound to a patient or subject. In the art, there are many methods of administering a compound, including, but are not limited to, intravenous, oral, aerosol, parenteral, intraocular, intrapulmonary, and topical administration.

[0392] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or carrier, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating material, that is involved in transporting or delivering a compound useful within the scope of this disclosure into or to a patient in order to perform its intended function. Typically, such a construct is transported or delivered from one organ or part of the body to another organ or part of the body. Each carrier must be compatible with other components of a formulation containing a compound useful within the scope of this disclosure and must be “acceptable” in the sense that it is not harmful to the patient. Some examples of materials that can function as pharmaceutically acceptable carriers include sugars, e.g., lactose, glucose, and sucrose; starches, e.g., corn starch and potato starch; cellulose and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; tragacanth powder; malt; gelatin; talc; excipients, e.g., cocoa butter and suppository waxes; oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, e.g., propylene glycol; polyols, e.g., glycerin, sorbitol, mannitol, and polyethylene glycol; esters, e.g., ethyl oleate and ethyl laurate; agar; buffers, e.g., magnesium hydroxide and aluminum hydroxide; surfactants; alginic acid; water free of pyrogens; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer; and other non-toxic, compatible substances used in pharmaceutical formulations.

[0393] As used herein, "pharmaceutically acceptable carrier" includes any coating agent, antibacterial and antifungal agent, and absorption delaying agent, etc. that is compatible with the activity of the compounds useful within the scope of the present disclosure and is physiologically acceptable to a patient. Auxiliary active compounds can also be incorporated into the composition. "Pharmaceutically acceptable carrier" may further include pharmaceutically acceptable salts of the compounds disclosed herein. Other additional components that can be included in the pharmaceutical composition are known in the art and are described, for example, in Remington’s Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA) (incorporated herein by reference).

[0394] The term "pharmaceutically acceptable", as used herein, refers to compounds, materials, compositions, and / or dosage forms that, within the scope of good medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic response, or other problems or complications and are commensurate with a reasonable benefit / risk ratio (e.g., in accordance with the guidelines of a governmental agency or other regulatory body, such as the U.S. Food and Drug Administration).

[0395] The phrase "pharmaceutically acceptable excipient", as used herein, refers to any component other than the active agent (e.g., as described herein) that is present in a pharmaceutical composition and has the property of being substantially non-toxic and non-inflammatory in a patient.

[0396] As used herein, the term "pharmaceutically acceptable salt" refers to derivatives of the disclosed cyclic peptides in which the parent compound is modified by converting any acidic or basic moieties present into their salt forms. In some embodiments, the side chain amino acid groups of the cyclic peptide (e.g., R 0 、R 1 、R 2 、R 3 、R 4These may be modified. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic salts of basic residues such as amines; and alkali or organic salts of acidic residues such as carboxylic acids. Examples of pharmaceutically acceptable salts in this disclosure include conventional non-toxic salts of the parent compound formed, derived from non-toxic inorganic or non-toxic organic acids. The pharmaceutically acceptable salts in this disclosure can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or free base form of these compounds with a stoichiometric amount of a suitable base or acid in water or an organic solvent, or a mixture thereof (generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred). In some embodiments, the pharmaceutically acceptable salt includes a corresponding pharmaceutically acceptable cation or anion. The term “pharmaceutically acceptable salt” is not limited to monosalts or 1:1 salts. For example, “pharmaceutically acceptable salt” also includes bissalts, such as bishydrochloride. A list of suitable salts, each of which is incorporated herein by reference as a whole, Remington's Pharmaceutical Sciences, 17 th This can be found in ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977).

[0397] As used herein, the terms “sample” or “biological sample” refer to a subset of its tissues, cells, or constituent parts (including, but not limited to, body fluids such as blood, serum, plasma, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, amniotic fluid, and urine). A sample may further include homogenates, lysates, or extracts prepared from a whole organism, or a subset of its tissues, cells, or constituent parts, or fragments or parts thereof, including, but not limited to, plasma, serum, cerebrospinal fluid, lymph, skin, external sections of the respiratory, intestines, and genitourinary tracts, tears, saliva, milk, blood cells, tumors, and organs. A sample may further refer to a culture medium, such as a nutrient broth or gel, which may contain cellular components or other biological materials, such as proteins (e.g., antibodies) or nucleic acid molecules.

[0398] As used herein, the term “Subject” refers to any entity to which a particular process or activity relates or applies. A Subject may include any living organism. Typical subjects include, but are not limited to, animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans) and / or plants.

[0399] As used herein, the term “target” refers to an object or entity affected by an action, or to an activity associated with a drug directed towards an object or entity (e.g., a drug that “targets” an object or entity). In some embodiments, a target refers to an antigen, epitope, or other structure to which an antibody or other compound is bound, or which is selected and / or used in the design, development, or isolation of an antigen-specific antibody or other compound. Targets may include, but are not limited to, nucleic acids, peptides, proteins, haptens, receptors, carbohydrates, glycans, enzymes, lipids, cells, and molecular structures comprising any of the aforementioned fragments or complexes.

[0400] When used to refer to the activity of a drug directed towards a target or entity, the term “target” may be used to describe the binding activity between a drug (e.g., an antibody or related structure) and such a target or entity (e.g., an antigen or epitope). For example, an antibody that binds to a specific antigen may be described as “targeting” a particular antigen or “directed” towards a particular antigen. Similarly, a compound (e.g., a targeted construct) that exhibits activity (e.g., therapeutic activity or cytotoxic activity) against a particular cell or tissue may be described as “targeting” the cell or tissue.

[0401] The target may include cells (referred to herein as “target cells”). Target cells may be in vivo or in vitro. Examples of target cells include blood cells, lymphocytes, cells lining the inside of the digestive tract such as oral and pharyngeal mucosa, cells forming the villi of the small intestine, cells lining the inside of the large intestine, cells lining the inside of the respiratory system (nasal cavity / lungs) of animals, dermal / epidermal cells, vaginal and rectal cells, visceral cells, placental cells, and blood-brain barrier cells. In some embodiments, target cells may be cancer cells, including, but are not limited to, those found in leukemia or tumors (e.g., tumors of the brain, lungs (small cell and non-small cell), ovaries, prostate, breast and colon, and other carcinomas and sarcomas). In yet other embodiments, target cells may be part of a tissue. Tissue having target cells or other target structures is referred to herein as target tissue. Target tissues are not limited to, but may include nerve tissue, intestinal tissue, pancreatic tissue, liver tissue, kidney tissue, prostate tissue, ovarian tissue, lung tissue, bone marrow tissue, and breast tissue.

[0402] As used herein, the term “target site” refers to a precise region on or within a target on which a given effector acts. A target site may be a precise region or epitope recognized by an antibody or compound. In some embodiments, a target site may be present only on one or more monomers of a polymeric structure (e.g., nucleic acids, peptides, polysaccharides, etc.). A target site may be formed by junctions or overlapping regions between two or more monomers or compounds.

[0403] As used herein, the term “therapeutic dose” means the amount of an agent that, when administered to a person suffering from or susceptible to a disease, disorder, and / or condition, is considered sufficient to treat or improve the symptoms of the disease, disorder, and / or condition, to diagnose, prevent, and / or delay the onset of the disease, disorder, and / or condition.

[0404] As used herein, terms such as “to treat” and “to cure” refer to any action taken to provide relief or mitigation of a pathological process. When relating to any of the therapeutic indications listed herein, terms such as “to treat” and “to cure” mean to reduce or alleviate at least one symptom associated with such indication, or to delay or prevent the progression or expected progression of such indication.

[0405] As used herein, the terms “prevent” or “prevent” mean that if the onset of a disability or disease has not occurred, there will be no onset of the disability or disease, or, if the onset of a disability or disease has already occurred, there will be no further onset of the disability or disease. The individual’s ability to prevent some or all of the symptoms associated with the disability or disease is also taken into consideration.

[0406] As used herein, the term “contact” refers to bringing together the indicated portions in an in vitro or in vivo setting. For example, “contact” cells with a compound includes administering the compound of the present invention to an individual, subject, or patient such as a human, and introducing the compound into a sample containing, for example, a purified preparation containing cells.

[0407] As used herein, the term “cell” means in vitro, ex vivo, or in vivo cells. In some embodiments, ex vivo cells may be part of a tissue sample excised from an organism such as a mammal. In some embodiments, in vitro cells may be cells in a cell culture. In some embodiments, in vivo cells are cells living in an organism such as a mammal.

[0408] As used herein, the term “tumor” refers to a group of cells that form in solid tissue as a result of abnormal cell proliferation and division. A benign or “non-cancerous” tumor remains isolated, while a malignant or “cancerous” tumor includes cells that can proliferate into the surrounding tissue.

[0409] As used herein, the term “tumor cells” refers to cells associated with or derived from a tumor. Benign or “non-cancerous” tumor cells remain associated with the tumor, while malignant or “cancerous” tumor cells can proliferate into the surrounding tissue.

[0410] As used herein, the following abbreviations are defined by the structures in Table D.

[0411] [Table 32]

[0412] [Table 33]

[0413] [Table 34]

[0414] [Table 35]

[0415] [Table 36]

[0416] [Table 37]

[0417] [Table 38]

[0418] Table 39

[0419] Table 40

[0420] Table 41

[0421] Table 42

[0422] Table 43

[0423] Table 44

[0424] Table 45

[0425] Table 46

[0426] Table 47

[0427] Table 48

[0428] [Table 49]

[0429] V. Equivalents and Scope While various embodiments of the disclosure have been specifically shown and described in this disclosure, it will be understood by those skilled in the art that various modifications of form and detail can be made without departing from the spirit and scope of the embodiments disclosed herein and set forth in the appended claims.

[0430] Those skilled in the art will recognize, or can verify by ordinary experimentation, many equivalents to the specific embodiments described herein. The scope of this disclosure is not intended to be limited to the foregoing description, but rather as set forth in the appended claims.

[0431] In the claims, articles such as “a,” “an,” and “the” may mean one or more unless otherwise clearly indicated by the context or unless otherwise stated. A claim or description containing “or” between one or more members of a group is deemed satisfied if one, two or more, or all, members of that group are present in, used in, or otherwise related to a given product or process, unless otherwise clearly indicated by the context or unless otherwise stated. This disclosure includes embodiments in which exactly one member of a group is present in, used in, or otherwise related to a given product or process. This disclosure includes embodiments in which two or more, or all, members of a group are present in, used in, or otherwise related to a given product or process.

[0432] Furthermore, it should be noted that the term “comprising” is intended to be open and allows, but does not require, the inclusion of additional elements or processes. Therefore, wherever the term “comprising” is used herein, the terms “consisting of” and “or including” are also included and disclosed.

[0433] Where a range is given, it includes the endpoint. Furthermore, unless otherwise indicated, or unless it is obvious from the context and the understanding of those skilled in the art, the values ​​expressed as a range may be any specific value or subrange within the ranges described in different embodiments of this disclosure, up to one-tenth of the lower limit unit of the range, unless otherwise specifically stated in the context.

[0434] Furthermore, it should be understood that any particular embodiment of the present disclosure contained in the prior art may be expressly excluded from one or more of the claims. Such embodiments may be excluded even if no such exclusion is explicitly stated herein, as they are considered to be known to those skilled in the art. Any particular embodiment of the compositions disclosed herein may be excluded from one or more of the claims for any reason, whether or not it relates to the existence of the prior art.

[0435] All cited sources, such as references, publications, databases, database entries, and technologies cited herein, are incorporated by reference to this application, even if not explicitly stated in the citation. In the event of any conflict between the cited sources and the descriptions in this application, the descriptions in this application shall prevail.

[0436] The headings of the sections and tables are not intended to be limiting. [Examples]

[0437] The compounds and methods disclosed herein are further illustrated by the following examples, which should not be construed as further limitations. Unless otherwise indicated, the implementation of this disclosure will involve the use of prior art in organic synthesis, cell biology, cell culture, and molecular biology, which are within the scope of the art.

[0438] Reagent - abbreviation ACN (or MeCN): Acetonitrile Ac2O: Acetic anhydride Acetic acid (ATOH) Boc:tert-butoxy-carbonyl DCM: Dichloromethane DIC: N,N'-Diisopropylcarbodiimide DIPEA: N,N-diisopropylethylamine DMF: N,N-dimethylformamide DMSO: Dimethyl sulfoxide Et2O: Diethyl ether Fmoc: Fluorenylmethyloxycarbonyl HATU:2-(3H-[1,2,3]triazolo[4,5-b]pyridine-3-yl)-1,1,3,3-tetramethylisouronium HOBt: 1-hydroxybenzotriazole MeOH: methanol Mpe:3-methylpentan-3-yl MTBE: Methyl tert-butyl ether NMP:N-methyl-2-pyrrolidone Oxyma (or OxymaPure): Cyanohydroxyiminoethyl acetate Dde: 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl TFA: Trifluoroacetic acid TIPS: Triisopropylsilane Cp*RuCl(PPh3)2: Chloro(pentamethylcyclopentadienyl)bis(triphenylphosphine)ruthenium(II) Grubbs2: (1,3-Dimethytylumidazolidine-2-ylidene)(Tricyclohexylphosphine)Benzylideneruthenium dichloride

[0439] Amino acids and constituent units ·Na-(((9H-fluoren-9-yl)methoxy)carbonyl)-1-(tert-butoxycarbonyl)-L-tryptophan(W) ·(((9H-fluoren-9-yl)methoxy)carbonyl)-L-threonine(T) ·(((9H-fluoren-9-yl)methoxy)carbonyl)-L-isoleucine(I) N-(((9H-fluoren-9-yl)methoxy)carbonyl)-S-trityl-L-cysteine(C) ·(S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(tritylamino)propanoic acid(N) ·Na-(((9H-fluoren-9-yl)methoxy)carbonyl)-Np-(tert-butoxycarbonyl)-L-histidine(H) ·(S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((3-methylpentan-3-yl)oxy)-4-oxobutanoic acid (D) ·(((9H-fluoren-9-yl)methoxy)carbonyl)-L-proline(P) ·N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N6-(1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl)-L-lysine(K(Dde)) ·N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N6-(1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl)-D-lysine(D-Lys(Dde)) ·N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N6-(tert-butoxycarbonyl)-D-lysine(D-Lys(Boc)) ·(S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(naphthalen-2-yl)propanoic acid (2Na) ·2-(4,7,10-Tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetic acid (DOTA(tBu)3) ·2,2',2”-(10-(2-((2,5-dioxopyrrolidine-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (DOTA-NHS) ·(S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-6,6-dimethyl-5-oxoheptanoic acid (gE) ·3-((tert-butoxycarbonyl)amino)propanoic acid (Boc-βAla) ·(S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)penta-4-enoic acid (allyl G) (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-azidopropanoic acid (DapN3) ·(S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)penta-4-ic acid (Pra)

[0440] Example 1. Preparation of Targeted Structures The targeted construct is prepared by combining a targeting moiety with a cargo. The targeting moiety incorporates a peptide sequence specific to a cancer cell antigen selected from DLL3. The cargo contains a radioactive agent including a radioisotope. The targeting moiety and cargo are combined using a linker.

[0441] In some embodiments, the DOTA chelating agent may be conjugated to the cyclic peptide targeting moiety according to the following general method.

[0442] General peptide synthesis for SEQ ID NOs: 3-89 (0.1 mmol scale): Peptides were synthesized using a CEM Liberty Blue microwave peptide synthesizer. Standard Fmoc chemistry and coupling were used with ProTide Rink amide resin (0.19 g / mmol).

[0443] Step 1: Deprotection: 20% piperidine in DMF (3 mL, 75 equivalents) was added to the resin, and the mixture was heated to 90°C for 1 minute. Next, the resin was washed four times with 4 mL of DMF.

[0444] Step 2: Double Coupling: Fmoc-protected amino acids (5 equivalents) in DMF (0.2M), DIC (10 equivalents) in DMF (1M), and OxymaPure (5 equivalents) in DMF (1M) were added to the microwave reaction vessel. Next, the microwave reaction vessel was heated to 90°C for 4 minutes and then discharged. Fmoc-protected amino acids (5 equivalents) in DMF (0.2M), DIC (10 equivalents) in DMF (1M), and OxymaPure (5 equivalents) in DMF (1M) were added to the reaction vessel. Next, the microwave reaction vessel was heated to 90°C for 4 minutes and then discharged.

[0445] Step 3: Steps 1 and 2 were repeated for the remaining amino acids listed in Table 4 (amino acid numbers 2-12; also referred to herein as "AA numbers 2-12").

[0446] Step 4: DOTA single coupling (amino acid number 13): The standard deprotection protocol (Step 1) used to remove the Fmoc group on AA number 12. 5 equivalents of 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetic acid in 0.2 M DMF, 10 equivalents of DIC in 1 M DMF, and 5 equivalents of OxymaPure in 1 M DMF were added to the microwave reaction vessel. The microwave reaction vessel was then heated to 50°C for 10 minutes and then discharged.

[0447] Step 5. Capping (AA No. 14; if DOTA chelating agent is not bound): The usual deprotection protocol (Step 1) is used to remove the Fmoc group on AA No. 12. After four DMF washes, 2.5 mL of 10% Ac2O in DMF is added to the microwave reaction vessel and then heated to 65°C for 2 minutes. The resin is then washed four times with 4 mL of DMF.

[0448] [Table 50]

[0449] Peptide cleavage, cyclization, and purification: Using the CEM Razor peptide cleavage system, peptide resins were transferred to frit-cleaved tubes, and 8 mL of cleavage solution (TFA / EDT / H2O / TIS, 94 / 2.5 / 2.5 / 1, v / v / v / v) was added to each tube. The peptide resins in the cleavage solution were heated at 40°C for 40 minutes.

[0450] Each tube is filtered, and the filtrate is collected in a centrifuge tube.

[0451] Next, precipitate the peptide with cold ether (45 mL). Then, centrifuge the precipitated peptide and rotate for at least 10 minutes. Next, decant the ether from each tube, leaving the peptide at the bottom. Then, repeat this ether precipitation process.

[0452] Next, the peptide was dissolved in 15 mL of MeCN / H2O (1 / 1, v / v), and 0.1 M I2 in MeOH was added dropwise until the yellow color persisted. The mixture was stirred at 20°C for 10 minutes. The reaction was quenched by adding 0.1 M Na2S2O3 dropwise until the yellow color disappeared. The reaction solution was then concentrated under reduced pressure. Next, for purification, the crude cyclized peptide was dissolved in 2 mL of DMSO.

[0453] The crude peptide in DMSO was directly purified by preparative HPLC (acidic conditions, TFA), followed by lyophilization to obtain the product.

[0454] [Table 51]

[0455] Example 2. Synthesis Protocol Synthesis Procedure B Example: Compound No. 18 [ka] Solid-phase peptide synthesis: The peptides were synthesized by standard solid-phase peptide synthesis (SPPS) using Fmoc / t-Bu chemistry. Assembly was performed on ProTide Rink amide resin (0.1 mmol scale, 0.19 g / mmol) on a CEM Liberty Blue microwave peptide synthesizer.

[0456] Fmoc deprotection was performed by adding 20% ​​(v / v) piperidine in DMF (3 mL) to the resin, and the mixture was heated to 90°C for 1 minute. Next, the resin was washed four times with DMF.

[0457] Amide coupling: Fmoc-protected amino acids (5 equivalents), DIC (10 equivalents) in DMF (1M), and OxymaPure (5 equivalents) in DMF (1M) were added to a microwave reaction vessel. The microwave reaction vessel was then heated to 90°C for 4 minutes and then discharged. For double coupling, this process was repeated twice.

[0458] The Fmoc deprotection and double coupling cycle was performed for all positions except DOTA(tBu)3, which was performed with triple coupling.

[0459] Peptide cleavage, cyclization, and purification: At the end of assembly, the resin was washed with DMF and DCM. Using a CEM Razor peptide cleavage system, the peptide was cleaved from the resin using 8 ml of cleavage solution (TFA / DODT / H2O / TIS, 92.5 / 2.5 / 2.5 / 2.5, v / v / v / v) for 30 minutes at 40°C. The resin was then filtered, and the peptide was precipitated with cold ether (45 ml). The precipitated peptide was then centrifuged for at least 5 minutes. The ether was decanted, leaving the peptide at the bottom of the centrifuge tube. This ether precipitation process was then repeated.

[0460] Next, the peptide was dissolved in 15 mL of MeCN / H2O (1 / 1, v / v), and 0.1 M I2 in MeOH was added dropwise until the yellow color persisted. The mixture was shaken at 20°C for 10 minutes. The reaction was quenched by adding 0.1 M Na2S2O3 dropwise until the yellow color disappeared. The reaction solution was then concentrated under reduced pressure. Next, for purification, the crude cyclized peptide was dissolved in 2 mL of DMSO.

[0461] Crude peptides were purified by preparative HPLC using a preparative Waters XBridge C18 column (100 × 19 mm, 5 μm OBD). Mobile phase A: H2O + 0.1% TFA; Mobile phase B: ACN + 0.1% TFA. The following gradient for eluent B was used: 20% B at 0–0.5 min; 20%–40% B at 0.5–14 min; 40–95% B at 14–15 min; held at 95% B at 15–17 min; 20% B at 17–18 min. Column temperature: ambient temperature. Flow rate: 24 mL / min. The recovered fractions were lyophilized to obtain the desired product.

[0462] Step C Example: Compound No. 19 [ka] Solid-phase peptide synthesis (SPPS) Representative peptides were synthesized manually using standard Fmoc chemistry on a 0.2 mmol scale.

[0463] Rink amide methylamine resin (0.20 mmol, 0.40 g, 0.50 mmol / g) was used for the synthesis. The resin was absorbed into DMF (5 mL) under nitrogen at room temperature for 2 hours. The supernatant was drained, and the resin was washed with DMF (3 × 3 mL, 30 seconds each).

[0464] Fmoc deprotection was performed by adding 20% ​​(v / v) piperidine in 10 mL of DMF to the resin, followed by washing with 5 × 10 mL of DMF for 30 seconds each.

[0465] Amide coupling was performed under nitrogen at room temperature for 30 minutes using Fmoc-protected amino acids / HATU / DIPEA (3, 2.85, and 6 equivalents, respectively). After the reaction was complete, the supernatant was drained and the resin was washed with DMF (5 × 10 mL for 30 seconds each). Completion of coupling was monitored by the ninhydrin test.

[0466] The Fmoc deprotection and amide coupling cycle was repeated with the desired monomer until a fully linear peptide was formed.

[0467] Upon completion of SPPS, the resin was first washed with DMF (5 × 20 mL, 30 seconds each), then washed with MeOH (3 × 20 mL, 30 seconds each) at room temperature, and finally dried under reduced pressure.

[0468] Peptide cleavage and cyclization The linear resin-bound peptide was deprotected and cleaved from the resin by treatment with (TFA / 3-MPA / TIS / H2O, 90 / 5 / 2.5 / 2.5, v / v / v, total 20 mL, where 3-MPA is 3-mercaptopropionic acid) for 2 hours at room temperature. After filtration of the resin, the crude linear peptide was precipitated from the cleavage solution using ice-cold isopropyl ether (80 mL) and recovered by centrifugation (3000 rpm, 2 minutes). The crude peptide was washed with ice-cold isopropyl ether (2 × 80 mL), and the residue was dried under reduced pressure at room temperature for 2 hours. Next, the crude product was dissolved in acetonitrile / water (1:2, v / v, 200 mL), and iodine in MeOH (0.1 M) was added dropwise at room temperature until the yellow color persisted. The mixture was stirred for a further 2 minutes at room temperature. Sodium thiosulfate in water (0.1 M) was added dropwise until the yellow color disappeared. The reaction mixture was freeze-dried to obtain a crude peptide.

[0469] Peptide purification The crude peptide was dissolved in 50% TFA in water and purified by preparative HPLC using a Gilson GX-281 system on back-to-back columns (Gemini, 5 μm, C18, 110 Å, along with Luna, 10 μm, C18, 100 Å). Mobile phase: (A) 0.075% TFA in H2O and (B) acetonitrile; flow rate: 20 mL / min; UV wavelength: 214 / 254 nm; gradient: 20-40% over 50 minutes; oven temperature: 30°C. The fractions with the desired mass (confirmed by LC / MS) were combined and lyophilized to obtain the desired product in TFA salt.

[0470] The peptide HCl salt form was obtained by dissolving the TFA salt peptide in 10% acetonitrile / water. The solution was desalted by preparative HPLC using a Gilson GX-281 system with back-to-back columns (Gemini, 5 μm, C18, 110 Å, along with Luna, 10 μm, C18, 100 Å). Mobile phase: (A) 0.05% HCl in H2O and (B) acetonitrile; flow rate: 20 mL / min; UV wavelength: 214 / 254 nm; gradient: first neutralize with 10 M NH4Cl (in H2O) for 25 minutes, then desalt with 0.05% HCl (in H2O) for 10 minutes, and finally elute with 0.05% HCl (in H2O) / ACN over 25 minutes with a gradient of 20–35%; oven temperature: 30°C. The fractions having the desired mass (confirmed by LC / MS) were combined and freeze-dried to obtain the final product in the HCl salt (78.6 mg, 18.8% yield, 98.2% purity) as a white solid.

[0471] The purity of the fraction was confirmed by UPLC. Column: Gemini C18, 5 μm, 110A, 150 × 4.6 mm. Mobile phase: (A) 0.1% TFA in water and (B) 0.075% TFA in acetonitrile; flow rate: 1 mL / min; UV wavelength: 220 nm.

[0472] Procedure L Step A. Solid-phase peptide synthesis The peptides were synthesized by standard solid-phase peptide synthesis (SPPS) using Fmoc / t-Bu chemistry. Assembly was performed on Rink-amide MBHA resin (250 μmol, 100-200 mesh; packed 0.42 mmol / g) on ​​a Cem Liberty Blue microwave peptide synthesizer (CEM Inc.). During peptide assembly on solid phase, the side chain protecting groups were as follows: tert-butyl for T; trityl for C and N; Boc for W and H; and Mpe for D.

[0473] All amino acids were dissolved in DMF at a concentration of 0.2 M. The acylation reaction was carried out at 90°C for 2 minutes under MW irradiation using activated amino acids in a 4-fold excess compared to the amino groups without resin. The amino acids were activated with an equimolar 1 M solution of DIC in DMF and a 1 M solution of Oxyma in DMF.

[0474] The double acylation reaction is T 2 H 8 , and W 11 We conducted the following:

[0475] DOTA was manually incorporated at room temperature using equimolar solutions of DOTA(tBu)3, DIC, and HOBt (3 equivalents, 1:1:1) in NMP, and complete acylation was monitored by the ninhydrin test.

[0476] At the end of assembly, the resin was washed with DMF, DCM, and Et2O. The peptide was cleaved from the solid support using 20 ml of TFA solution (v / v: 87.5% TFA, 5% H2O, 2.5% TIPS, 5% phenol) at room temperature for approximately 4 hours. The resin was then filtered, concentrated to approximately 5 mL, and precipitated in cold MTBE (45 mL). After centrifugation, the peptide pellet was washed with fresh cold Et2O to remove organic scavengers. This process was repeated twice. The final pellet was dried, resuspended in H2O and ACN 1:1, stirred overnight, and then lyophilized to obtain crude linear peptides.

[0477] Step B. Cyclization with thioacetal linker The linear peptide (210 mg) from step A was dissolved in H2O / ACN (1 mg / mL). TCEP HCl (3 equivalents), followed by diiodomethane (50 equivalents) and DIPEA (0.5% v / v) were added. The mixture was stirred at room temperature. The reaction was completed after 3 hours (monitored by ULC-MS). The mixture was quenched with TFA and lyophilized.

[0478] Crude peptides were purified by reverse-phase HPLC in two runs using a preparative Waters XBridge C18 column (150 × 50 mm, 130 Å, 5 μm). Mobile phase A: H2O + 0.1% TFA; Mobile phase B: ACN + 0.1% TFA. The following gradient for eluent B was used: from 20% B to 20% B over 5 minutes, and to 35% B over 25 minutes, at a flow rate of 80 mL / min and a wavelength of 214 nm. The recovered fractions were lyophilized to obtain the desired peptides as TFA salts.

[0479] A pure peptide was dissolved in H2O / ACN (1 mg / mL), and a 50 mM solution of HCl (10 equivalents for each amino group) was added. The resulting solution was freeze-dried, then redissolved in H2O / ACN (1 mg / mL), and freeze-dried again to obtain the final thioacetal cyclization product.

[0480] General Procedure N Peptides were synthesized using a CEM Liberty Blue microwave peptide synthesizer. Standard Fmoc chemistry and coupling were used with Wang resin (0.75 mmol / g).

[0481] Wang resin was manually filled using the following protocol. The Wang resin was transferred to a frit syringe and swollen with DMF. 4 equivalents of Fmoc-protecting amino acids, 4 equivalents of HOBT, 4 equivalents of DIC, and 1 equivalent of DMAP were added to the frit syringe in a total of 5 mL of DMF. The reaction mixture was shaken for 4 hours. The reaction mixture was filtered and washed with DMF (4 × 5 mL, 30 seconds each) and DCM (4 × 5 mL, 30 seconds each). The resin was then dried under reduced pressure.

[0482] Step 1. Deprotection: 20% piperidine (3 mL, 75 equivalents) in DMF was added to the resin, and the mixture was heated to 90°C for 1 minute. Next, the resin was washed with DMF (4 × 4 mL).

[0483] Step 2. Double Coupling: Fmoc-protected amino acids (5 equivalents), DIC in DMF (1M) (10 equivalents), and OxymaPure in DMF (1M) (5 equivalents) were added to the microwave reaction vessel. Next, the microwave reaction vessel was heated to 90°C for 4 minutes and then discharged. Fmoc-protected amino acids (5 equivalents), DIC in DMF (1M) (10 equivalents), and OxymaPure in DMF (1M) (5 equivalents) were added to the reaction vessel. Next, the microwave reaction vessel was heated to 90°C for 4 minutes and then discharged.

[0484] Step 3. Steps 1 and 2 were repeated for the remaining amino acids.

[0485] Step 4. Capping: The standard deprotection protocol (Step 1) used to remove the Fmoc group on the final amino acid. After washing with DMF (4 × 4 mL), 2.5 mL of 10% Ac2O in DMF was added to the microwave reaction vessel and then heated to 65°C for 2 minutes. The resin was then washed with DMF (4 × 4 mL).

[0486] Step 5. Orthogonal deprotection: The resin was washed with DMF (2 × 4 mL). 4 mL of 2% hydrazine in DMF was added to the microwave reaction vessel and mixed at room temperature for 30 minutes. Next, the resin was washed with DMF (5 × 4 mL).

[0487] Step 6. DOTA Coupling: The final DOTA coupling is added under triple coupling conditions. DOTA(tBu)3 (5 equivalents), DIC (10 equivalents) in DMF (1M), and OxymaPure (5 equivalents) in DMF (1M) were added to the microwave reaction vessel. The microwave reaction vessel was then heated to 90°C for 4 minutes and then discharged. This process was repeated two more times.

[0488] At the end of the assembly, the resin was cleaned with DMF and DCM.

[0489] Using a CEM Razor peptide cleavage system, peptides were cleaved from the resin using 8 ml of cleavage solution (TFA / DODT / H2O / TIS, 92.5 / 2.5 / 2.5 / 2.5, v / v / v / v) at 40°C for 30 minutes. The resin was then filtered, and the peptides were precipitated with cold ether (45 ml). The precipitated peptides were then centrifuged for at least 5 minutes. The ether was decanted, leaving the peptides at the bottom of the centrifuge tube. This ether precipitation process was then repeated. The peptides were then dissolved in 10 mL of MeCN / H2O (1 / 1, v / v) and freeze-dried.

[0490] Crude linear peptides were dissolved in 25 mL of ACN / H2O (1:1). DODT (2 equivalents, 32 μL) was added to the reaction mixture, followed by DIPEA (250 μL-400 μL) to adjust the reaction solution pH to 8-10. Finally, diiodomethane (20 equivalents, 150 μL) was added to the reaction solution, and the mixture was shaken at room temperature for 2 hours. The reaction was monitored by HPLC, and once the reaction was complete, the reaction was quenched with 100 μL of TFA. The reaction solvent was removed by lyophilization, and the sample was purified by HPLC.

[0491] Crude peptides were purified by preparative HPLC using a preparative Waters XSelect Peptide CSH C18 column (1.9 cm × 25 cm inner diameter (5 μm / 130 Å)). Mobile phase A: 20 mM NH4HCO3 in water; Mobile phase B: ACN. Gradient: 20% B at 0-1 min; 20%-50% B at 1-35 min; 50-90% B at 35-36 min; 90-95% B at 36-36.5 min; 95% B at 36.5-40 min; 95-20% B at 40-41 min; 20% B at 41-45 min. Column temperature: ambient temperature. Flow rate: 24 mL / min. The recovered fractions were freeze-dried to obtain the desired product.

[0492] Table 5 below shows the synthetic procedures used to synthesize the compounds described herein.

[0493] [Table 52]

[0494] [Table 53]

[0495] Table 6 below shows the LCMS analysis of the compounds described herein.

[0496] [Table 54]

[0497] [Table 55]

[0498] Example 3. Binding affinity of the targeted partial peptide to DLL3 Affinity determination by surface plasmon resonance (SPR). SPR tests were performed on the compounds disclosed herein using the following protocol.

[0499] procedure: Binding affinity and binding kinetics parameters at 25°C were determined using Biacore S200 or Biacore 8K instruments. The biotinylated target ligand human DLL3 was immobilized on a streptavidin sensor chip. A reference surface was set up for nonspecific binding and refractive index changes. For analysis of interaction kinetics, samples of various concentrations were injected at a flow rate of 100 μL / min using a running buffer containing 20 mM phosphate buffer with 2.7 mM KCl, 137 mM NaCl, 0.05% surfactant P20, and 2% DMSO. Sensorgram curves were fitted to a 1:1 binding model, and affinity and kinetics parameters were obtained using Biacore Insight evaluation 4.0 software.

[0500] Table 7 below shows the K values ​​obtained by SPR assay for the selected group of compounds. DThe values ​​are shown. In this table, "A" represents K. D Represents ≤ 1.0 nM; "B" is 1.0 nM <K D Represents ≤10nM; "C" is 10nM <K D Represents ≤100nM; "D" is 100nM <K D This represents ≤300nM.

[0501] [Table 56]

[0502] [Table 57]

[0503] Example 4. Radiolabeling and ex vivo in vivo distribution of targeted peptides [ 177 General procedure for radiolabeling of chelating agents-peptides with Lu]LuCl [ 177 Lu]LuCl3 was obtained from a vendor in HCl solution. It was added to the reaction vial (1.5 mL Eppendorf vial). 177 To all mCi in Lu]LuCl3, ammonium acetate buffer (0.2 M, pH 4.9, 100 μL, containing 1% w / v ascorbic acid and 6% v / v ethanol) and a peptide conjugate (1 nmol, also referred to herein as "chelating agent-peptide") were added. The peptide conjugate of this disclosure is also referred to herein as "chelating agent-peptide". The pH of the solution was determined to be approximately 5 by using a pH strip. The reaction vial was incubated at 80°C, 700 RPM for 17-20 minutes. Samples from the reaction were analyzed by RP-HPLC using an Agilent Infinity II 1260 HPLC to determine the completion of the reaction and [ 177The radiochemical purity of the Lu]Lu-chelating peptide was determined. HPLC conditions: Waters XBridge BEH C18 column, 130 Å, 3.5 μm, 4.6 mm × 250 mm; mobile phase: solvent A = water (containing 0.1% formic acid), solvent B = acetonitrile (containing 0.1% formic acid). Gradient: 0.5 mL / min flow rate, 25–45% B at 10 min, 45–65% B at 12 min, 65–90% at 6 min. The required amount of product was mixed in 1% PBSA for cell testing.

[0504] [ 225 General procedure for radiolabeling of chelating agents-peptides with Ac]Ac The peptide conjugate of this disclosure is reacted in a reaction mixture containing an excipient and an acetate buffer or equivalent buffer with ethanol. 225 Radiolabeling with Ac]Ac isotope. Heat the reaction mixture to achieve radiolabeling, for example, by heating the reaction mixture at 90°C for 15 minutes. Radiolabeling [ 225 The Ac]Ac-chelating agent-peptide is further diluted to the desired radioactive concentration in a formulation buffer containing additional excipients. Radiolabeled [ 225 DTPA was added to the sample of the Ac]Ac-chelating agent-peptide product, and the analysis was performed by RP-HPLC, with fractions collected every 12 seconds. 225 After a time-dependent equilibrium is achieved between Ac]Ac and its daughter isotopes (>6 hours after collection), the fraction is analyzed using gamma spectroscopy, and the resulting CPM is plotted as a function of time. Radiochemical purity (%RCP) data is obtained from this reconstitution chromatography.

[0505] [Table 58]

[0506] [Table 59]

[0507] General protocols for ex vivo in vivo distribution studies Tumor volume in xenograft mice is 200-400 mm². 3 If within the specified range, and if the mice were 5-7 weeks old for the tumor-untreated mouse study, a bolus dose of radiolabeled peptide was intravenously injected into the mice (via the lateral tail vein) for the ex vivo in vivo distribution study. To evaluate the amount of radionuclide injected into individual mice, [ 177 The syringe was assayed with a dose calibrator before and after injection of the Lu]Lu peptide.

[0508] Mice were euthanized at predetermined time points, selected tissues were excised, and collected in pre-weighed tubes. The weight of the tubes was then re-measured after excision, and the difference represented the weight of each tissue. Radioactivity in each tissue was measured using a gamma counter. The counts were corrected for decay over injection time, and the percentage of injected radioactivity per gram (%Ia / g) was calculated for each tissue based on the radioactivity injected into each individual mouse. The injected radioactivity was converted to counts based on the sensitivity of the gamma counter. Next, the counts in the tissue were converted to a percentage of injected radioactivity, and this was divided by the mass of the tissue to obtain %Ia / g.

[0509] [Table 60]

[0510] The subject matter disclosed should not be limited in scope by the specific embodiments and examples described herein. In fact, various modifications of this disclosure, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description and the accompanying drawings. Such modifications are intended to be included within the scope of the accompanying claims.

[0511] All references cited herein (e.g., publications, patents, or patent applications) are incorporated herein by reference in whole for all purposes to the same extent that each individual reference (e.g., publications, patents, or patent applications) is specifically and individually indicated to be incorporated in whole for all purposes. Other embodiments are within the scope of the following claims.

Claims

1. Cyclic peptide of formula B: 【Chemistry 1】 or its pharmaceutically acceptable salt (In the formula, P １ is, -L １ - Chelating agents, 【Chemistry 2】 Selected from; D １ It is a -NR''- chelating agent; L １ It does not exist, or 【Transformation 3】 Selected from; Here, L １ The amino group is P １ Alternatively, it can bond to the carbonyl group of the chelating agent to form an amide bond; P ２ Yes, C(O)NH ２ ,C(O)OH, 【Chemistry 4】 Selected from; D ２ is OH or NH ２ ; and L ２ It does not exist, or 【Transformation 5】 Selected from; Here, L ２ The amino group is P ２ It links to the carbonyl group to form an amide bond; P ３ H, 【Transformation 6】 Selected from; L ３ It does not exist, or independently, 【Transformation 7】 Selected from; Here, L ３ The carbonyl group is P ２ It links to the amine group to form an amide bond; D ３ CH is independent. ３ , C(O)OH, and 【Transformation 8】 Selected from; X is a halogen, R ０ This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R １ This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R ２ This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R ３ This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; B １ C １～６ It is alkylene; C １ C １～６ It is alkylene; A １ teeth, 【Chemistry 9】 Selected from; Here, w is selected from 1, 2, or 3; R ４ These are amino acid side chains of natural amino acids or amino acid side chains of unnatural amino acids, and both are CH ２ C(O)OH or C(O)(CH ２ CH ２ O) ｐ (CH ２ ) ２ N(CH ３ ) ３ ＋ It is arbitrarily replaced by; R ５ This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R ６ This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R ７ teeth, (i) amino acid side chains of natural amino acids, (ii) Selected from the amino acid side chains of unnatural amino acids, or (iii) 【Chemistry 10】 Selected from the group consisting of; R ８ This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R ９ This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; R １０ This refers to the amino acid side chain of a natural amino acid or the amino acid side chain of a non-natural amino acid; m is either 0 or 1; Each n, q, and u is an integer between 0 and 16, independently of the others; Each p is an integer between 0 and 24, independently; Each s is an independent integer between 0 and 16; Each t is independently 1, 2, 3, 4, 5, or 6; Each R' independently consists of H, C(O)OH, and (CH ２ ) Selected from OH and NHAc; Each R'' independently corresponds to H and CH ３ Selected from; Here, the variable base R ０ , R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R ９ , or R １０ However, when defined as an amino acid side chain of a cyclic amino acid, the corresponding amino acid nitrogen in the peptide backbone of the general formula forms part of the cyclic group; The aforementioned cyclic peptide may optionally contain a radionuclide.

2. The cyclic peptide of formula B is the cyclic peptide of formula Ia: 【Chemistry 11】 The cyclic peptide according to claim 1, or a pharmaceutically acceptable salt thereof.

3. The cyclic peptide of formula B is the cyclic peptide of formula Ib: 【Chemistry 12】 The cyclic peptide according to claim 1, or a pharmaceutically acceptable salt thereof.

4. P １ teeth, 【Chemistry 13】 , -L １ - Chelating agents, and 【Chemistry 14】 Selected from; Here, D １ It is an NR'' chelating agent; L １ It does not exist, or 【Chemistry 15】 And; n and s are independently integers between 2 and 15; A cyclic peptide according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein p is 8, 9, 10, 11, or 12.

5. P １ is, -L １ - A chelating agent, a cyclic peptide according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof.

6. P １ DOTA, 【Chemistry 16】 A cyclic peptide according to any one of claims 1 to 3, selected from, or a pharmaceutically acceptable salt thereof.

7. P ２ is C(O)NH ２ A cyclic peptide according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein the peptide is C(O)OH.

8. P ２ Yes, C(O)NH ２ ,C(O)OH, and 【Chemistry 17】 A cyclic peptide according to any one of claims 1 to 3, selected from, or a pharmaceutically acceptable salt thereof.

9. P ２ Yes, C(O)NH ２ ,C(O)OH, [Chemistry 18] Selected from; P ３ is, Ac or 【Chemistry 19】 And; X is a halo, the cyclic peptide according to claim 8, or a pharmaceutically acceptable salt thereof.

10. The cyclic peptide of formula B is the cyclic peptide of formula II: 【Chemistry 20】 or its pharmaceutically acceptable salt (In the formula, P ２ is C(O)NH ２ (or C(O)OH) The cyclic peptide according to claim 1, or a pharmaceutically acceptable salt thereof.

11. m is 0; P １ DOTA, 【Chemistry 21】 Selected from; P ２ Yes, C(O)NH ２ ,C(O)OH, 【Chemistry 22】 Selected from; P ３ is, Ac or 【Chemistry 23】 And; X is a halo; R １ , Trp, 2Na, 1Na, 4CF ３ Selected from the group consisting of amino acid side chains of -Phe, 7-Aza-Trp, 1Me-Trp, 5OH-Trp, BIP, 5OMe-Trp, 4F-Phe, 3Pya, 4Pya, PAF, MAF, OAF, 5Qui, 7MeO-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, D-Ala, Ala, α-Me-Trp, and NMe-Trp; R ２ This is selected from the group consisting of amino acid side chains of Thr, D-Ala, Ala, α-Me-Thr, Lys, and NMe-Thr; R ３ are Ile, Env, CHA, CBA, Nle, Tbg, THPG, Chg, 2Nal, 1Nal, 2CF ３ Selected from the group consisting of amino acid side chains of -Phe, 2PhEt-Ala, D-Ala, Ala, Leu, t-Bu-Ala, NMe-Nle, α-tert-amyl Gly, Allo-Ile, Lys(C12), Lys(C14), Lys(C16), α-Me-Ile, and NMe-tBuAla; A １ teeth, 【Chemistry 24】 Selected from the group consisting of; R ４ is Asn, D-Ala, Ala, DAB-4-NHCOC ５ H １１ , DAB-4-NHCOC ７ H １５ Asp, Ser, Lys, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, 3Pya, 4Pya, Glu, NMee-Asn, PipA (acetic acid), and Pip(PegNMee) ３ ) Selected from the group consisting of amino acid side chains of Ala; R ５ is selected from the group consisting of the amino acid side chains of Asn, Ala, D-Ala, Trp, Asp, Lys, 3Pya, 4Pya, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, Glu, NMe-Asn, and Ser; R ６ , is Trp, 4CF ３ Selected from the group consisting of amino acid side chains of -Phe, 1Me-Trp, 7Aza-Trp, BIP, 2Naal, 1Naal, α-Me-Trp, D-Ala, Ala, 4F-Phe, 5F-Trp, 5Ome-Trp, Asn, 5OH-Trp, 7Me-Trp, 7MeO-Trp, 7Cl-Trp, and NMe-Trp; R ７ teeth, (i) selected from the group consisting of amino acid side chains of 3Pya, 4Pya, Lys(Me)3, His, Ala, D-Ala, Gln, Lys, Glu, Arg, Orn, NMe-His, and Ser; or (ii) 【Chemistry 25】 Selected from the group consisting of; Each s is independently 3, 5, 10, 12, or 14; R ８ This is selected from the group consisting of amino acid side chains of Asp, D-Ala, Ala, Asn, Thr, NMe-Asp, and α-Me-Asp; R ９ This is selected from the group consisting of amino acid side chains of Trp, 7-aza-Trp, 1Me-Trp, D-Ala, Ala, 4F-Phe, 1Naal, 2Naal, 5F-Trp, 5Ome-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp; R １０ The cyclic peptide according to claim 1, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of amino acid side chains of Pro, D-Ala, Ala, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, Pip, 5,5-di-Me-Pro, NMe-Ser, trans4NH2-Pro, cis4NH2-Pro, Sar, Aze, NMe-Ala, NMe-Leu, R-3Me-Aze, α-Me-Aze, ACI, and 3Me2-Aze.

12. m is 1; P １ is DOTA, 【Chemistry 26】 Selected from; P ２ Yes, C(O)NH ２ ,C(O)OH, 【Chemistry 27】 Selected from; P ３ This is Ac and 【Chemistry 28】 Selected from; X is a halo; R ０ This is selected from the group consisting of amino acid side chains of Gly, Met, D-Ala, Ala, Nle, and Nva; R １ , Trp, 2Na, 1Na, 4CF ３ Selected from the group consisting of amino acid side chains of -Phe, 7-aza-Trp, 1Me-Trp, 5OH-Trp, BIP, 5Ome-Trp, 4F-Phe, 3Pya, 4Pya, PAF, MAF, OAF, 5Qui, 7MeO-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, D-Ala, Ala, α-Me-Trp, and NMe-Trp; R ２ This is selected from the group consisting of amino acid side chains of Thr, D-Ala, Ala, α-Me-Thr, Lys, and NMe-Thr; R ３ are Ile, Env, CHA, CBA, Nle, Tbg, THPG, Chg, 2Nal, 1Nal, 2CF ３ Selected from the group consisting of amino acid side chains of -Phe, 2PhEt-Ala, D-Ala, Ala, Leu, t-Bu-Ala, NMe-Nle, α-tert-amyl Gly, Allo-Ile, Lys(C12), Lys(C14), Lys(C16), α-Me-Ile, and NMe-tBuAla; A １ teeth, 【Chemistry 29】 Selected from the group consisting of; R ４ is Asn, D-Ala, Ala, DAB-4-NHCOC ５ H １１ , DAB-4-NHCOC ７ H １５ Selected from the group consisting of amino acid side chains of Asp, Ser, Lys, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, 3Pya, 4Pya, Glu, and NMe-Asn; R ５ This is selected from the group consisting of amino acid side chains of Asn, Ala, D-Ala, Trp, Asp, Lys, 3Pya, 4Pya, 3-(4-piperidinyl)-Ala, 3-(1-morpholinyl)-Ala, Glu, NMe-Asn, and Ser; R ６ , Trp, 4CF ３ Selected from the group consisting of amino acid side chains of -Phe, 1Me-Trp, 7Aza-Trp, BIP, 2Naal, 1Naal, α-Me-Trp, D-Ala, Ala, 4F-Phe, 5F-Trp, 5Ome-Trp, Asn, 5OH-Trp, 7Me-Trp, 7MeO-Trp, 7Cl-Trp, and NMe-Trp; R ７ is selected from the group consisting of amino acid side chains of 3Pya, 4Pya, Lys(Me)3, His, Ala, D-Ala, Gln, Lys, Glu, Arg, Orn, NMe-His, and Ser; or R ７ teeth, 【Transformation 30】 Selected from the group consisting of; Each s is independently 3, 5, 10, 12, or 14; R ８ This is selected from the group consisting of amino acid side chains of Asp, D-Ala, Ala, Asn, Thr, NMe-Asp, and α-Me-Asp; R ９ This is selected from the group consisting of amino acid side chains of Trp, 7-aza-Trp, 1Me-Trp, D-Ala, Ala, 4F-Phe, 1Naal, 2Naal, 5F-Trp, 5Ome-Trp, α-Me-Trp, 7Cl-Trp, 5OH-Trp, 7Me-Trp, 7MeO-Trp, and NMe-Trp; R １０ The cyclic peptide according to claim 1 or 3, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of amino acid side chains of Pro, D-Ala, Ala, α-Me-Pro, trans4-fluoro-Pro, cis4-fluoro-Pro, trans4OH-Pro, cis4OH-Pro, Pip, 5,5-di-Me-Pro, NMe-Ser, trans4NH2-Pro, cis4NH2-Pro, Sar, Aze, NMe-Ala, NMe-Leu, R-3Me-Aze, α-Me-Aze, ACI, and 3Me2-Aze.

13. B １ CH ２ or C(CH ３ ) ２ And; C １ CH ２ or C(CH ３ ) ２ A cyclic peptide according to any one of claims 1 to 9, 11, and 12, or a pharmaceutically acceptable salt thereof.

14. B １ CH ２ And; C １ CH ２ A cyclic peptide according to any one of claims 1 to 9, 11, and 12, or a pharmaceutically acceptable salt thereof.

15. The chelating agents are, independently, ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 1,4,7,10-tetra-azacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), 6-((16-((6-carboxypyridine-2-yl)methyl)-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane-7-yl)methyl)-4-isothiocyanatopicolinic acid (Macropa), Macrodipa, 2,2',2'',2'''-(1,10-dioxa-4,7,13,16-tetraazacyclooctadecane-4,7,13,16-tetrayl)tetraacetic acid) (Crown), 1 A cyclic peptide according to any one of claims 1 to 14, selected from the group consisting of 4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, α-(2-carboxyethyl) (DOTAGA), 1,4,7-triazacyclononane-N,N',N''-triacetic acid (NOTA), 1,4,7,10-tetraazacyclododecane-N,N',N'',N''-tetraacetic acid (TETA), 1,4,7,10,13-pentazacyclopentadecane-N,N',N'',N''',N''-pentaacetic acid (PEPA), and 1,4,7,10,13,16-hexaazacyclohexadecane-N,N',N'',N'',N'',N''''-hexaacetic acid (HEHA), or a pharmaceutically acceptable salt thereof.

16. Formula B is a cyclic peptide according to any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, wherein formula B is substituted with at least one chelating agent.

17. The cyclic peptide of formula B is a cyclic peptide according to any one of claims 1 to 16, selected from the cyclic peptides in Table A, or a pharmaceutically acceptable salt thereof.

18. The cyclic peptide of formula B is a cyclic peptide according to any one of claims 1 to 16, selected from the cyclic peptides in Table B, or a pharmaceutically acceptable salt thereof.

19. The cyclic peptide is the cyclic peptide according to any one of claims 1 to 18, further comprising a radioisotope, or a pharmaceutically acceptable salt thereof.

20. The radioactive isotope is selected from the radioactive isotopes in Table 3, and the cyclic peptide according to claim 19, or a pharmaceutically acceptable salt thereof.

21. A pharmaceutical composition comprising a cyclic peptide according to any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

22. A method for treating cancer in a subject requiring treatment, comprising administering to the subject a cyclic peptide according to any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 21.

23. The method according to claim 22, wherein the cancer is a DLL3-mediated cancer.

24. The method according to claim 22 or 23, wherein the cancer is a neuroendocrine tumor, melanoma, or primary brain tumor.

25. The method according to claim 24, wherein the neuroendocrine tumor is selected from small cell lung cancer (SCLC), medullary thyroid carcinoma (MTC), large cell neuroendocrine carcinoma (LCNEC), pancreatic and gastrointestinal neuroendocrine carcinoma (GEPNEC), neuroendocrine prostate cancer (NEPC), small cell prostate cancer (SCPC), Merkel cell carcinoma (MCC), neuroendocrine cervical cancer, and grade 3 neuroendocrine tumor (NET).

26. A peptide having binding specificity to DLL3, wherein it binds to one or more amino acids A81, L83, G106, A85, and R61 of the DLL3 amino acid sequence of SEQ ID NO: 1, and the peptide has the amino acid sequence of formula A: 【Chemistry 31】 Peptides containing or pharmaceutically acceptable salts thereof (In the formula, X ０ is any natural or non-natural amino acid, or X ０ It is non-existent; X １ This is selected from Trp, 7-ather-Trp, 1Me-Trp, 5OH-Trp, 5OMe-Trp, 7OMe-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, α-Me-Trp, and NMe-Trp; X ２ and X ３ Each of these is independently any natural or non-natural amino acid; Y １ It is Cys; X ４ , X ５ , X ６ , X ７ and X ８ Each of these is independently any natural or non-natural amino acid; Y ２ It is Cys; X ９ This is selected from Trp, 7-ather-Trp, 1Me-Trp, 5OH-Trp, 5OMe-Trp, 7OMe-Trp, 7Me-Trp, 5F-Trp, 7Cl-Trp, α-Me-Trp, and NMe-Trp; X １０ This is selected from Pro, α-Me-Pro, trans4fluoro-Pro, cis4fluoro-Pro, trans4OH-Pro, cis4OH-Pro, 5,5-diMe-Pro, trans4NH2-Pro, and cis4NH2-Pro; P １ is, -L １ - Chelating agents, 【Chemistry 32】 Selected from; D １ It is a -NR''- chelating agent; L １ It does not exist, or 【Transformation 33】 Selected from; Here, L １ The amino group is P １ Alternatively, it can bond to the carbonyl group of the chelating agent to form an amide bond; Each n, q, and u is an integer between 0 and 16, independently of the others; Each p is an integer between 0 and 24, independently; Each s is an independent integer between 0 and 16; Each t is independently 1, 2, 3, 4, 5, or 6; Each R' independently consists of H, C(O)OH, and (CH ２ ) Selected from OH and NHAc; Each R'' independently corresponds to H and CH ３ Selected from; Here, the cyclic peptide is Y １ and Y ２ Cyclized via a linker between them; Here, the cyclic peptide binds to DLL3.

27. The peptide according to claim 26, which binds to amino acids A81, L83, G106, A85 and R61 of the DLL3 amino acid sequence of SEQ ID NO:

1.

28. The peptide according to claim 26 or 27, wherein it is bound to the main chain atoms of amino acids A81, L83, G106 and A85 of the DLL3 amino acid sequence of SEQ ID NO:

1.

29. The peptide according to any one of claims 26 to 28, wherein it is bound to the side chain atom of amino acid R61 of the DLL3 amino acid sequence of SEQ ID NO:

1.

30. The peptide according to any one of claims 26 to 29, wherein it is bound to the main chain atoms of amino acids A81, L83, G106 and A85 of the DLL3 amino acid sequence of SEQ ID NO: 1, and to the side chain atom of amino acid R61 of the DLL3 amino acid sequence of SEQ ID NO:

1.

31. Approximately 1×10 －８ M ~ approx. 1 x 10 －１０ M's SPR K Ｄ The peptide according to any one of claims 26 to 30, which can bind to DLL3 by value.

32. The peptide according to any one of claims 26 to 31, comprising the amino acid sequence of WTACANAKDCWP, or a derivative thereof containing one or more non-natural amino acids.

33. The peptide according to claim 32, wherein amino acids W1, A3, A7, and W11 are bound to DLL3.

34. A pharmaceutical composition comprising the peptide according to any one of claims 26 to 33 and a pharmaceutically acceptable carrier.

35. A method for treating cancer in a subject requiring treatment, comprising administering to the subject a peptide according to any one of claims 26 to 33 or a pharmaceutical composition according to claim 34.

36. The method according to claim 35, wherein the cancer is a DLL3-mediated cancer.

37. The method according to claim 35 or 36, wherein the cancer is a neuroendocrine tumor, melanoma, or primary brain tumor.

38. The method according to claim 37, wherein the neuroendocrine tumor is selected from small cell lung cancer (SCLC), medullary thyroid carcinoma (MTC), large cell neuroendocrine carcinoma (LCNEC), pancreatic and gastrointestinal neuroendocrine carcinoma (GEPNEC), neuroendocrine prostate cancer (NEPC), small cell prostate cancer (SCPC), Merkel cell carcinoma (MCC), neuroendocrine cervical cancer, and grade 3 neuroendocrine tumor (NET).

39. The method according to claim 24 or 37, wherein the neuroendocrine tumor is extrapulmonary neuroendocrine carcinoma (NEC) of the cervix.

40. The cyclic peptide according to claim 17, wherein the cyclic peptide is radiolabeled with F-18, Ga-68, In-111, Lu-177, or Ac-225, as well as pharmaceutically acceptable salts and solvates thereof.

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