Peptide receptor radionuclide therapy

Abstract

The present disclosure provides methods and related compositions utilizing peptide receptor radionuclide therapy (PRRT) and a theranostic regimen in conjunction with a molecular targeting regimen via integrin subtypes α v β6.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 625,717, filed January 26, 2024, which is incorporated herein by reference in its entirety.

[0003] sequence list

[0004] The text of the sequence list in computer-readable form, titled "UCDA_42815_601_SequenceListing.xml" and 8,574 bytes in size, which was generated on January 24, 2025, is hereby incorporated in its entirety by reference. Background Technology

[0005] Cancer continues to demand new and more effective treatments. In particular, some cancer types and subtypes are resistant to current approaches. For example, the incidence of pancreatic ductal adenocarcinoma (PDAC) continues to rise, with an estimated 47,000 new cases diagnosed in the United States in 2018. Unfortunately, PDAC remains the deadliest cancer, with 98% of patients ultimately dying from the disease. Despite extensive testing and some encouraging progress in first- and second-line treatments, only one chemotherapy therapy (gemcitabine) has been found to offer some benefit for this disease. However, the clinical response rate is quite low (approximately 30%), and even lower in advanced cases (see, for example, Andriulli A., et al., Ann. Surg. Oncol. 2011;19:1644-1662; Hashimoto K., et al., Oncology. 2009;77:217-223). This lack of clinical options for pancreatic cancer suggests that the need for research into new therapies is clearly unmet. Summary of the Invention

[0006] On the one hand, this disclosure provides therapeutic conjugates of formula I or formula IV: (Formula I) (Formula IV) Wherein RN is a radionuclide; where X is 5G or ABM-5G; where 5G is a peptide; and where ABM is the albumin-binding portion.

[0007] In some implementations, X is ABM-5G, such that the resulting therapeutic conjugate is included in Formula II: (Formula II).

[0008] In some implementations, X is 5G, such that the resulting therapeutic conjugate is included in Formula III: (Formula III)

[0009] Such therapeutic conjugates are not limited to a specific chemical moiety of RN. In some embodiments, RN is... 67 Cu. In some implementations, RN is selected from... 47 Sc、 67 Cu、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 212 Pb, 223 Ra and 225 Ac. In other examples, the RN used for the conjugate has imaging capabilities (such as in diagnostic applications), and the RN is selected from... 11 C 13 N、 15 O、 18 F, 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 68 Ga、 82 Rb、 86 Y、 111 In、 124 I and 125 I. In some examples, the therapeutic conjugate may also be visualized after administration. The therapeutic conjugate is not limited to 5G as a specific chemical moiety of the peptide. In some embodiments, the peptide is configured to bind α v β6 integrin. In some embodiments, the peptide is configured to bind α. vRGD peptide of β6 integrin. In some embodiments, the peptide is a PEGylated peptide. In some embodiments, the PEGylated peptide has the following amino acid sequence: GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), and includes a first PEG moiety at the N-terminus of the peptide and a second PEG moiety at the C-terminus of the peptide. In some embodiments, the first PEG moiety and the second PEG moiety are independently selected from PEG. 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and / or (PEG 28 )2 (PEG 1500×2). In some embodiments, the first PEG portion and the second PEG are the same. In some embodiments, the first PEG portion and the second PEG each contain PEG. 28 (PEG 1500).

[0010] Such therapeutic conjugates are not limited to a specific chemical moiety of the albumin-binding portion (ABM). In some embodiments, the ABM is 4-(4-iodophenyl)butyric acid or the K(D-Abu-iodophenylbutyryl) moiety.

[0011] Such therapeutic conjugates are not limited to specific uses or functions. In some embodiments, therapeutic conjugates of formula I, II, III, or IV are used to deliver radioactivity to a patient or subject. In some embodiments, therapeutic conjugates of formula I, II, III, or IV are used to treat or prevent cancer in a patient or subject. In some embodiments, therapeutic conjugates of formula I, II, III, or IV are used to treat... v 6. Integrin-related cancers.

[0012] In related aspects, this disclosure provides for the treatment v 6. Methods for treating integrin-related cancers, which include administering one or more doses of a therapeutic conjugate of formula I, II, III, or IV to a subject requiring treatment.

[0013] In some embodiments, one or more doses include administering one dose. In some embodiments, one or more doses include administering two or three additional doses (e.g., four, five, six, ten, twenty, fifty, or one hundred) of the therapeutic conjugate to the subject.

[0014] Such methods are not limited to a specific amount of one or more doses of the therapeutic conjugates of Formula I, II, III, or IV. In embodiments where multiple doses of the therapeutic conjugate are administered to a subject, each dose may contain the same or different amounts of radioactivity and / or the same or different amounts of peptides.

[0015] This includes administering a dose of a therapeutic conjugate of formula I, II, III, or IV to a subject requiring treatment to treat α. v Such methods for treating β6 integrin-related cancers are not limited to a specific dose or dose type of the treatment conjugate. In some embodiments, the dose is a radioactive dose. In some embodiments, the dose of the treatment conjugate of Formula I, II, III, or IV contains about 25 mCi to about 200 mCi of radioactivity. In some embodiments, the amount of radioactivity in the dose is about 25 mCi, 50 mCi, 100 mCi, 150 mCi, or 200 mCi.

[0016] This method is not limited to a specific type or category. v 6. Integrin-related cancers. In some implementations, the aforementioned... v 6. Integrin-related cancers include pancreatic cancer, breast cancer, colorectal cancer, lung cancer, ovarian cancer, cervical cancer, oral squamous cell carcinoma, skin squamous cell carcinoma, gastric cancer, or endometrial cancer. In some embodiments, the... v 6. Integrin-associated cancers are locally advanced, unresectable, or metastatic forms of cancers such as pancreatic cancer, breast cancer, colorectal cancer, lung cancer, ovarian cancer, cervical cancer, oral squamous cell carcinoma, cutaneous squamous cell carcinoma, gastric cancer, and endometrial cancer. In some implementations, pancreatic cancer is locally advanced or metastatic pancreatic cancer; locally advanced, unresectable, or metastatic pancreatic adenocarcinoma; or pancreatic ductal adenocarcinoma (PDAC).

[0017] In some implementations, the v 6. Integrin-associated cancers include primary and metastatic lesions. In some embodiments, the... v 6. Integrin-related cancers include lesions in the adrenal glands, bone, brain, liver, lungs, or any combination thereof.

[0018] In some embodiments, the subject receives standard care treatment prior to administration of the therapeutic conjugate dose. In some embodiments, the subject receives standard care treatment after administration of the therapeutic conjugate dose. In some embodiments, the subject receives standard care treatment both before and after administration of the therapeutic conjugate dose. In some embodiments, the standard care treatment includes one or more of surgery, radiation therapy, chemotherapy, chemoradiotherapy, and targeted therapy. In a specific embodiment, the standard care treatment includes the FOLFIRINOX regimen (leucovorin, fluorouracil, irinotecan hydrochloride, and oxaliplatin), gemcitabine, abraxane, irinotecan, or a combination thereof.

[0019] In some embodiments, the method further includes scanning the body or portions thereof of the subject after application of the therapeutic conjugate. In some embodiments, the scan includes positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), or single photon emission computed tomography (SPECT).

[0020] In some embodiments, the method further includes administering a diagnostic conjugate prior to the administration of the therapeutic conjugate, wherein the diagnostic conjugate comprises an integrin-binding moiety (such as an RGD peptide) and a second radionuclide (such as a different radionuclide suitable for imaging). In some embodiments, the second radionuclide is... 68 Ga. In some embodiments, the diagnostic conjugate is administered at a dose containing up to about 5 mCi (e.g., greater than about 0.01 mCi to about 5 mCi) of radioactivity.

[0021] In some embodiments, the therapeutic conjugate is administered within 5 weeks after the administration of the diagnostic conjugate.

[0022] In some embodiments, the method further includes administering an amino acid solution to the subject. In some embodiments, the solution is administered both before and simultaneously with the administration of the therapeutic conjugate.

[0023] In some embodiments, the therapeutic conjugate is administered to the subject by infusion. In some embodiments, the diagnostic conjugate is administered to the subject by injection. In some embodiments, the diagnostic conjugate is administered by bolus, slow bolus, or slow infusion. In some embodiments, the therapeutic conjugate is administered by bolus, slow bolus, or slow infusion. In some embodiments, the diagnostic conjugate is administered by bolus, and the therapeutic conjugate is administered by slow bolus or slow infusion. In some embodiments, the therapeutic conjugate is infused over several minutes or hours, for example, about 1 minute, about 2 minutes, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 55 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, or about 8 hours. In a specific embodiment, the therapeutic conjugate is infused over about 30 minutes. In some embodiments, the diagnostic conjugate is infused over several minutes, such as about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, or less. In a specific embodiment, the diagnostic conjugate is infused over about 5 minutes or less.

[0024] In some embodiments, the treatment results in disease stabilization, partial remission, or complete remission. In some embodiments, the treatment results in a reduction of cancer metastasis in the subject. In some embodiments, the treatment results in a reduction in the volume, size, or growth of the tumor in the subject. In some embodiments, the treatment results in an increased response of the cancer to subsequently administered anticancer agents.

[0025] In some embodiments, the amount of therapeutic conjugate present in kidney tissue approximately 24 hours, 48 ​​hours, or 72 hours after administration is lower than the amount of therapeutic conjugate present in kidney tissue 1 hour after administration. In some embodiments, the ratio of the amount of therapeutic conjugate in the primary tumor to the amount of therapeutic conjugate in kidney tissue approximately 24 hours, 48 ​​hours, or 72 hours after administration is higher than the ratio of the amount of therapeutic conjugate in the primary tumor to the amount of therapeutic conjugate in kidney tissue 1 hour after administration.

[0026] In another aspect, this disclosure provides pharmaceutical compositions comprising peptides having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1).

[0027] In some embodiments, the PEG moiety is covalently linked to the N-terminus, C-terminus, or both of the N-terminus and C-terminus of the peptide. In some embodiments, the PEG moiety is independently selected from PEG. 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and / or (PEG 28 )2 (PEG 1500×2). In other embodiments, the first PEG portion is covalently linked to the N-terminus of the peptide, the second PEG portion is covalently linked to the C-terminus of the peptide, and the first and second PEG portions are independently selected from PEG. 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and / or (PEG 28 )2 (PEG 1500×2). In some examples, the first PEG portion and the second PEG portion are the same. In a specific embodiment, the first PEG portion and the second PEG portion each contain PEG 28 (PEG 1500). In some embodiments, the PEG moiety covalently linked to the C-terminus of the peptide is terminated with an amide, carboxyl, or hydroxyl group.

[0028] In some embodiments, the peptide is covalently linked to an albumin-binding moiety (ABM). In a specific embodiment, the ABM comprises 4-(4-iodophenyl)butyric acid. In some embodiments, the ABM comprises a linker, such as a peptide linker covalently linked to the peptide, a first PEG moiety, or a second PEG moiety. In some embodiments, the ABM comprises a K(D-Abu-iodophenylbutyryl) moiety.

[0029] In some embodiments, the peptide is covalently linked to the chelating moiety. In a specific embodiment, the chelating moiety is NOTA (2,2',2”-(1,4,7-triazacyclononane-1,4,7-triyl)triacetic acid) or DOTA (2,2',2”,2”'-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid).

[0030]

[0031] (2,2',2”,2”'-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid)

[0032] In some examples, a radionuclide (RN) is complexed with the chelate portion. In some embodiments, the radionuclide is covalently linked to the peptide directly or indirectly. In some examples, the RN is selected from... 47 Sc、 67 Cu、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 203 Pb, 212 Pb 223 Ra and 225 Ac. In other examples, the RN is selected from 11 C 13 N、 15 O、 18 F, 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 111 In、 124 I, 125 I, 131 I, 203 Pb. In other examples, the RN is 177 Lu or 68 Ga. In other examples, the RN is 67 Cu or 64 Cu.

[0033] In related aspects, this disclosure provides pharmaceutical compositions comprising conjugates of formula I or formula IV. (Formula I) (Formula IV) RN and X are as described in this article. Attached Figure Description

[0034] Figure 1 Integrin α was shown to be kept at 37°C for 1 hour.v Cell binding and internalization in β6(+)Capan-1 cells (n=3 / compound); Column: SD. Peptide 1 = DOTA-ABM-5G ( ); Peptide 2 = NOTA-ABM-5G ( ).

[0035] Figure 2 This demonstrates the injection of imaging peptides. 64 Cu-1 and 64 Representative axial (top) and coronal (bottom) PET / CT images of mice bearing Capan-1 tumors (yellow arrows) obtained after Cu-2. K - kidney, L - liver. PET color scale bar.

[0036] Figures 3A to 3B It shows that the charge has α v Imaging peptides in mice with β6(+)Capan-1 tumors, expressed as a percentage of the injected dose per gram of tissue (%ID / g). 64 Cu-1 (A) and 64 Biodistribution of Cu-2(B) (n=3 / group / time point).

[0037] Figures 4A to 4C It shows the comparison with current radiotherapy 177 Lu-DOTA-ABM-5G ( 177 Compared to Lu-1), 64 Cu-1, 64 Cu-2 uptake by (A) Capan-1 tumors, (B) kidneys, and (C) Capan-1 to kidney ratio. Peptide 1 = DOTA-ABM-5G; Peptide 2 = NOTA-ABM-5G.

[0038] Figure 5 The control group received 37 MBq of the first dose on day 0. 67 Cu-1, or four doses on days 0, 7, 14, and 28 (37 MBq per dose). 67 The average tumor volume of Cu-1 mice from day 0 to day 42. Detailed Implementation

[0039] I. Introduction

[0040] This article presents a combination of peptide receptor radionuclide therapy (PRRT) and a therapeutic approach via integrin subtype α. v Methods and related compositions for molecular targeting of β6.

[0041] Integrin subtype α vβ6 is an epithelial-specific cell surface receptor that is undetectable in healthy adult epithelial tissues but is significantly upregulated in various cancers of epithelial origin, including pancreatic ductal adenocarcinoma (PDAC). α v β6 was initially identified in PDAC, where almost all tumors showed α. v β6 was highly upregulated.

[0042] PRRT is a therapy that uses cell-targeting peptides in combination with radioactive isotopes. When injected into the patient's bloodstream, the radioactive peptides deliver a targeted high dose of radiation directly to cancer cells.

[0043] II. Definition

[0044] Unless otherwise expressly stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Furthermore, any methods and materials similar to or equivalent to those described herein may be used to practice this disclosure. For the purposes of this disclosure, the following terms are defined.

[0045] The terms “an,” “a,” or “described” as used herein include not only aspects of a single member but also aspects of more than one member. For example, the singular forms “an,” “a,” and “described” include plural referents unless the context clearly indicates otherwise. Thus, for example, references to “an adjuvant” include multiple such adjuvants, and so on.

[0046] The term "about" is used herein to modify numerical values ​​and to indicate a defined range around said numerical value. If "X" is a numerical value, then "about X" generally means a value between 0.90X and 1.10X. Any reference to "about X" means at least the numerical values ​​X, 0.90X, 0.91X, 0.92X, 0.93X, 0.94X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, 1.05X, 1.06X, 1.07X, 1.08X, 1.09X, and 1.10X. Thus, "about X" is intended to disclose, for example, "0.98X". When "about" is applied at the beginning of a range of numerical values, it applies to both ends of the range. When "about" is applied to the first value of a set of numerical values, it applies to all values ​​in that set.

[0047] The term "peptide" refers to a compound consisting of a single chain of D-amino acids or L-amino acids, or a mixture of D-amino acids and L-amino acids linked by peptide bonds. Typically, peptides are about 2 to about 50 amino acids in length. As a non-limiting example, peptides present in the conjugates described herein are about 5 to about 45 amino acids, about 8 to about 45 amino acids, about 8 to about 25 amino acids, about 8 to about 20 amino acids, about 12 to about 45 amino acids, about 12 to about 30 amino acids, or about 20 amino acids in length.

[0048] The term "RGD peptide" refers to peptides that can be integrin (such as α-peptides). v β6 integrin recognizes polypeptides containing an arginine-glycine-aspartic acid (Arg-Gly-Asp) tripeptide motif. In fact, as used herein, "RGD peptide" refers to a conjugate exhibiting a similarity to α-integrin in this document. v The ability of β6 integrin to specifically interact with binding / interacting peptide motifs. In some embodiments, the RGD peptide interacts with α... v β6 integrin interacts with and / or binds to molecules without cross-reacting with similar sequences or structures. In some cases, RGD peptides specifically bind to α-integrin when they bind with a substantially lower dissociation constant (i.e., tighter binding) than molecules with similar sequences or structures. v β6 integrin. For example, in some cases, when the RGD peptide has an affinity for α-integrin that is approximately 2, 3, 4, 5, 6, 8, 10, 15, 20, 25, 30, 40, 50, 100, or 1000 times higher than that of the related molecule. v β6 integrin binds specifically to β6. RGD peptide binds to α... v Binding of β6 integrins can occur via intermolecular forces such as ionic bonds, hydrogen bonds, hydrophobic interactions, dipole-dipole bonds, and / or van der Waals forces. Cross-reactivity can be assessed by evaluating the interaction between RGD peptides and α-integers under normal conditions. v The binding of β6 integrin and a variety of molecules that are more or less structurally and / or functionally related can be tested. These methods may include, but are not limited to: binding studies, blocking and competition studies with closely related molecules, FACS analysis, surface plasmon resonance (e.g., using BIAcore), analytical ultracentrifugation, isothermal titration calorimetry, fluorescence anisotropy, fluorescence spectroscopy, radiolabeled ligand binding assays, and combinations of the above methods.

[0049] As used herein, the term "polyethylene glycolation" refers to the process of covalently coupling a polyethylene glycol (PEG) molecule with other molecules, such as RGD peptides, which is subsequently referred to as "PEGylation." As a non-limiting example, RGD peptides can be PEGylated simultaneously at both the amino and carboxyl ends using monodisperse PEG molecules with defined chain lengths to produce bi-terminated PEGylated peptide conjugates. Monodisperse PEG molecules typically comprise discrete molecular weights with precisely defined numbers of repeating ethylene glycol units. Suitable PEG moieties are commercially available from Polypure AS (Oslo, Norway), which supplies monodisperse PEG molecules and their PEG derivatives consisting essentially of only one oligomer (e.g., oligomer purity greater than about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%). In a specific implementation, the RGD peptide is PEGylated at both ends with a single type of monodisperse PEG moiety or a mixture of different types of monodisperse PEG moiety, wherein the monodisperse PEG moiety has a molecular weight of less than about 5000 Daltons (Da) (e.g., less than about 5000 Da, 4000 Da, or 3000 Da), for example, PEG. 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and / or (PEG 28 )2(PEG 1500×2)).

[0050] The term "radioactive nuclide" is intended to include any nuclide that exhibits radioactivity. A "nuclide" refers to a type of atom defined by its atomic number, atomic mass, and energy state, such as carbon-14 (…). 14 C). “Radioactivity” refers to radiation emitted by radioactive materials, including alpha particles, beta particles, nucleons, electrons, positrons, neutrinos, and gamma rays. Examples of radionuclides suitable for use in the conjugates described herein include, but are not limited to, tritium ( 3 H), Fluorine-18 ( 18 F), Phosphorus 32 ( 32 P), sulfur 35 ( 35 S), Scandium-47 ( 47 Sc), Cobalt 55 ( 55 Co), Copper 60 ( 60 Cu), Copper 61 ( 61 Cu), Copper 62 ( 62 Cu), Copper 65 ( 64 Cu), Gallium-66 ( 66 Ga), Copper 67 ( 67 Cu), Gallium-67 ( 67 Ga), gallium 68( 68 Ga), Rubidium 82( 82 Rb), Yttrium 86 (86 Y), Yttrium 87 ( 87 Y), Strontium-89 ( 89 Sr), Strontium-90 ( 90 Sr), Yttrium 90 ( 90 Y), Rhodium 105 ( 105 Rh), Silver 111 ( 111 Ag), Indium 111 ( 111 In), Iodine-124 ( 124 I), iodine 125( 125 I), iodine 131( 131 I), Tin 117m ( 117 mSn), technetium 99m( 99m Tc), Cesium-137 137 Cs), Promethium-149 ( 149 Pm), Samarium-153 ( 153 Sm), Terbium 149 ( 149 Tb), Terbium 152 ( 152 Tb), Terbium 155 ( 155 Tb), Terbium 161 161 Tb), Holmium-166 ( 166 Ho), Lutetium 177( 177 Lu), Rhenium 186 ( 186 Re), Rhenium 188 ( 188 Re), Thallium-201 201 Tl), astatine 211( 211 At), astatine 215( 215 At), astatine 217( 217 At), astatine 218( 218 At), Bismuth 209 ( 209 Bi), bismuth 211( 211 Bi), bismuth 212( 212 Bi), bismuth 213( 213 Bi), lead 203( 203 Pb), Lead-212 ( 212 Pb), polonium-210 ( 210 Po), Polonium-211 211 Po), Polonium-212 212 Po), Polonium-214 214 Po), Polonium-215 215 Po), Polonium-216 216 Po), Polonium-218 218 Po), Radon 218( 218 Rn), Radon 219 ( 219 Rn), Radon 220 ( 220 Rn), Radon 222 ( 222Rn), Radon 226 ( 226 Rn), Francium-221 ( 221 Fr), Radium-223 ( 223 Ra), Radium-224 224 Ra), Radium-226 226 Ra), Actinium 225 ( 225 Ac), Actinium 227 ( 227 Ac), Thorium-227 ( 227 Th), Thorium-228 228 Th), Thorium-229 229 Th), Thorium-230 ( 230 Th), Thorium-232 ( 232 Th), Protactinium 231 ( 231 Pa), uranium-233 ( 233 U), uranium 234( 234 U), uranium 235( 235 U), uranium 236( 236 U), uranium 238( 238 U), Neptunium 237( 237 Np), Plutonium-238 238 Pu), Plutonium-239 239 Pu), Plutonium-240 240 Pu), Plutonium-244 244 Pu), Americium 241 ( 241 Am), Curium 244 ( 244 Cm), Curium 245 ( 245 Cm), Curium 248 ( 248 Cm), Californium-249 ( 249 Cf) and Californium-252 ( 252 Cf). As used in this article, 117m Sn and 99m The "m" in Tc stands for metastable state. Furthermore, naturally occurring radioactive elements such as uranium, radium, and thorium (often representing mixtures of radioactive isotopes) are also suitable examples of radionuclides. 67 Cu、 131 I, 177 Lu and 186 Re represents a radionuclide that emits beta and gamma. 212 Bi is a radionuclide that emits α and β. 226 Ra is a radionuclide that emits α and γ. 211 At、 215 At、 217 At、 218 At、 209 Bi、 211 Bi、 213 Bi、210 Po、 211 Po、 212 Po、 214 Po、 215 Po、 216 Po、 218 Po、 218 Rn、 219 Rn、 220 Rn、 222 Rn、 226 Rn、 221 Fr、 223 Ra、 224 Ra、 225 Ac、 227 Ac、 227 Th、 228 Th、 229 Th、 230 Th、 232 Th、 231 Pa、 233 U、 234 U、 235 U、 236 U、 238 U、 237 Np, 238 Pu、 239 Pu、 240 Pu、 244 Pu、 241 Am、 244 Cm、 245 Cm、 248 Cm、 249 Cf and 252 Cf is an example of a radionuclide that emits α. 3 H, 32 P, 35 S, 47 Sc、 89 Sr、 90 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 149 Pm, 153 Sm、 166 Ho、 188 Re and 212 Pb is an example of a radionuclide that emits β. 67 Ga、 111 In、 99m Tc, 137 Cs、 201 Tl and 203 Pb is an example of a radionuclide that emits γ. 55Co、 60 Cu、 61 Cu、 62 Cu、 66 Ga、 68 Ga、 82 Rb and 86 Y is an example of a radioactive nuclide that emits positrons. 64 Cu is a radioactive nuclide that emits both beta and positrons.

[0051] It should be noted that in U.S. Provisional Patent Application No. 63 / 625,717, filed January 26, 2024, which claims priority to this patent application, the abbreviation "Ra" is defined as a radionuclide in the general formula described herein. To avoid confusion with the abbreviation for the chemical element "radium" (e.g., Ra), the abbreviation "RN" is used in the general formula described herein to define a radionuclide. Therefore, "RN" as used herein is equivalent to "Ra" in the general formula described in U.S. Provisional Patent Application No. 63 / 625,717.

[0052] The term “object” or “patient” usually refers to humans, but can also include other animals, such as other primates, rodents, canines, felines, horses, sheep, pigs, etc.

[0053] As used herein, the term "application" includes oral administration, topical application, suppository administration, intravenous administration, intraperitoneal administration, intramuscular administration, intralesional administration, intrathecal administration, intrathecal administration, intranasal administration, or subcutaneous administration, or implantation of a sustained-release device, such as a micro-osmotic pump, into the subject. Routes of administration are any, including parenteral and transmucosal (e.g., oral, sublingual, palatal, gingival, nasal, vaginal, rectal, or percutaneous). Parenteral administration includes intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intravenous, and intracranial administration. Other delivery methods include, but are not limited to, the use of liposomal formulations, intravenous infusion, and transdermal patches. The term "co-application" means that the conjugate described herein is administered simultaneously with, before, or after the administration of a second agent.

[0054] III. RGD peptides and conjugates

[0055] It has been previously confirmed that this has been observed in the dose-limiting organ, the kidney¹ 77 Uptake of Lu-DOTA-ABM-5G. To reduce renal uptake and improve peptide pharmacokinetics, experiments conducted during the development of embodiments of the present invention resulted in the production of alternative radionuclides (for imaging). 64 Cu,t 1 / 2 =12.7 hours; and for treatment 67 Cu,t 1 / 2 =2.6 days) and different chelating agents (DOTA, NOOTA). Using64 Cu and 67 Cu was used to radiolabel two peptides, DOTA-ABM-5G(1) and NOA-ABM-5G(2), to produce 64 Cu / 67 Cu-DOTA-ABM-5G ( 64 Cu / 67 Cu-1) and 64 Cu / 67 Cu-NOTA-ABM-5G ( 64 Cu / 67 The cell binding and serum stability of Cu-2 were evaluated in vitro, and in vivo evaluation was conducted using PET / CT imaging and biodistribution studies.

[0056] Therefore, this article provides therapeutic conjugates (“conjugates”) for peptide receptor radionuclide therapy (PRRT), and related PRRT methods using such therapeutic conjugates, wherein the conjugates are configured to bind α v β6 integrin.

[0057] In some implementations, selective binding α is provided. v A peptide conjugate of β6 integrin. In some embodiments, the peptide selectively binds to α. v RGD peptide of β6 integrin. In some embodiments, the peptide comprises the RGD motif RGDLX1X2X3 (SEQ ID NO: 2), wherein X1 and X2 are independently selected amino acids, and X3 is L or I. In some embodiments, the RGD peptide does not contain any alanine residues. In some embodiments, the RGD peptide is 8 to 40 amino acids. In some cases, the RGD peptide is more than 20 amino acids. In some cases, the RGD peptide is 21 amino acids. In some embodiments, the RGD peptide further comprises QX4VX5RT (SEQ ID NO: 7) located at the C-terminus of the RGD motif, wherein X4 is R or K, and X5 is A or G. In some cases, the RGD peptide comprises the amino acid sequence QRVGRT (SEQ ID NO: 3) located at the C-terminus of the RGD motif. In some cases, the RGD peptide comprises the amino acid sequence RGDLQVLGQRVGRT (SEQ ID NO: 4). In some embodiments, the RGD peptide comprises the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1). In some embodiments, the RGD peptide consists of, or is substantially composed of, the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1).

[0058] In some embodiments, the conjugate further comprises one or more polyethylene glycol (PEG) moieties covalently linked to the peptide (e.g., the RGD peptide). In some cases, the conjugate comprises two PEG moieties, for example, one PEG moiety covalently linked to the N-terminus of the peptide and another PEG moiety covalently linked to the C-terminus of the peptide (e.g., the RGD peptide). In some embodiments, the conjugate comprises the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), having a first PEG moiety covalently linked to the N-terminus of the peptide and a second PEG moiety covalently linked to the C-terminus of the peptide. In some embodiments, the PEG moiety covalently linked to the C-terminus of the peptide is terminated with an amide, carboxyl, or hydroxyl group.

[0059] In some embodiments, the first PEG portion and the second PEG portion each have a molecular weight of less than about 5000 Daltons (Da), for example, less than about 3000 Da. In some embodiments, the first PEG portion and the second PEG portion are monodisperse PEG portions having a defined chain length. Non-limiting examples of PEG portions having a defined chain length include small monodisperse PEG molecules having an oligomer purity greater than about 95%. In some examples, the first PEG portion and the second PEG portion are independently selected from PEG. 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and (PEG 28 )2 (PEG 1500×2). In a specific embodiment, the first PEG portion and the second PEG portion are the same. In some embodiments, both the first PEG portion and the second PEG portion are PEG. 28 (PEG 1500). In some embodiments, the conjugate comprises the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), wherein the N-terminus and C-terminus of the peptide each have a PEG covalently linked thereto. 28 (PEG 1500) portion (also referred to herein as "5G"). In some embodiments, PEG is covalently linked to the C-terminus of the peptide. 28 The (PEG 1500) portion is terminated with an amide group, a carboxyl group, or a hydroxyl group.

[0060] In some embodiments, the conjugate comprises an albumin-binding moiety (ABM) covalently linked to the conjugate. The ABM may increase the half-life of the conjugate in serum, for example, when administered to a subject. In some embodiments, the ABM is covalently linked to the peptide (e.g., the RGD peptide). In some embodiments of conjugates having one or more PEG moieties, the ABM is covalently linked to the peptide (e.g., the RGD peptide), a first PEG moiety, or a second PEG moiety. In some embodiments, the ABM comprises a linker, such as a peptide linker covalently linked to the peptide, the first PEG moiety, or the second PEG moiety. In some embodiments, the ABM comprises 4-(4-iodophenyl)butyric acid (IPA), or a homologue thereof having a shorter alkyl chain, such as 4-(4-iodophenyl)propionic acid or 4-(4-iodophenyl)acetic acid, or the ABM comprises 4-(4-methylphenyl)butyric acid or 4-(4-bromophenyl)butyric acid, or a homologue thereof having a shorter alkyl chain, such as its propionic or acetic acid homologue. In some embodiments, the ABM is covalently linked to the first and / or second PEG moiety via a linker such as a glutamate (E) linker, a peptide linker such as a lysine-aspartic-aminobutyric acid (KD-Abu) linker, or other suitable linkers (such as amino acid or peptide linkers) well known to those skilled in the art. In some embodiments, the ABM comprises an ε-(4-(4-iodophenyl)butyramide)lysine-glutamate moiety (“K(IPA)E”), which corresponds to an IPA covalently linked to a side chain of a lysine residue of the lysine-glutamate peptide linker. In some other embodiments, the ABM comprises a K(D-Abu-iodophenylbutyryl) moiety, which corresponds to an IPA covalently linked to an aminobutyric acid residue of the lysine-aspartic-aminobutyric acid (KD-Abu) peptide linker. In some embodiments, the ABM comprising a K(IPA)E or K(D-Abu-iodophenylbutyryl) moiety is covalently linked to the first PEG moiety. In some embodiments, the conjugate comprises the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), wherein the N-terminus and C-terminus of the peptide each have a PEG moiety covalently linked thereto, such as PEG. 28 The conjugate contains a (PEG 1500) portion, and further comprises ABM covalently linked thereto. In some embodiments, the PEG portion covalently linked to the C-terminus of the peptide is terminated with an amide, carboxyl, or hydroxyl group.

[0061] In other embodiments, the imaging agent or therapeutic agent is covalently linked to the conjugate (e.g., via a prosthetic group, chelating agent, or linker). In some embodiments, the imaging agent or therapeutic agent is covalently linked to an albumin-binding motif covalently linked to a first PEG motif (e.g., via a prosthetic group, chelating agent, or linker) such that the imaging agent or therapeutic agent is the most N-terminal portion of the conjugate.

[0062] In some embodiments, the conjugate comprises a peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), wherein the N-terminus and C-terminus of the peptide each have a PEG moiety covalently linked thereto, such as PEG. 28 The (PEG 1500) portion, ABM is covalently linked to the PEG portion at the N-terminus of the peptide, and the therapeutic agent is covalently linked to the N-terminus of the conjugate. In some embodiments, the conjugate comprises a peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), wherein the N-terminus and C-terminus of the peptide each have a PEG portion covalently linked thereto, such as PEG 28 The (PEG 1500) portion comprises ABM covalently linked to the PEG portion at the N-terminus of the peptide, and a developer covalently linked to the N-terminus of the conjugate. In some embodiments, the PEG portion covalently linked to the C-terminus of the peptide is terminated with an amide, carboxyl, or hydroxyl group.

[0063] In some embodiments, the therapeutic conjugate comprises a therapeutic agent. In some embodiments, the therapeutic agent is a radionuclide, such as a radionuclide emitting α, β, and / or γ. In some embodiments, the therapeutic conjugates herein comprise, for example, ³H, ¹H. 8 F, ³²P, ³ 5 S, 47 Sc、 55 Co、 60 Cu、 6 ¹Cu、 6 ²Cu、 64 Cu、 66 Ga、 67 Cu、 67 Ga、 68 Ga、 8 ²Rb、 86 Y、 87 Y、 89 Sr、 90 Sr、 90 Y、¹ 05 Rh、¹¹¹Ag、¹¹¹In、¹² 4 I、¹² 5 I、¹³¹I、¹¹ 7m Sn、 99m Tc、¹³ 7 Cs、¹ 49 Pm、¹ 5 ³Sm、¹ 66 Ho、¹ 77 Lu、¹ 86 Re、¹88 Re、² 0 ¹Tl、²¹¹At、²¹ 5 At、²¹ 7 At、²¹ 8 At、² 09 Bi、²¹¹Bi、²¹²Bi、²¹³Bi、² 0 ³Pb、²¹²Pb、²¹ 0 Po,²¹¹Po,²¹²Po,²¹ 4 Po、²¹ 5 Po、²¹ 6 Po、²¹ 8 Po、²¹ 8 Rn、²¹ 9 Rn、²² 0 Rn,²²²Rn,²² 6 Rn,²²¹Fr,²²³Ra,²² 4 Ra、²² 6 Ra、²² 5 Ac、²² 7 Ac、²² 7 Th、²² 8 Th、²² 9 Th、²³ 0 Th,²³²Th,²³¹Pa,²³³U,²³ 4 U、²³ 5 U、²³ 6 U、²³ 8 U、²³ 7 Np,²³ 8 Pu、²³ 9 Pu、² 40 Pu、² 44 Pu、² 4 ¹Am、² 44 Cm,² 45 Cm,² 48 Cm,² 49 Cf and² 5 ²Cf radionuclides: . In some embodiments, the therapeutic conjugates described herein contain, for example, 47 Sc、 67 Cu、 89 Sr、 90 Y、¹ 05 Rh、¹¹¹Ag、¹¹ 7m Sn、¹³¹I、¹ 49 Pm、¹ 5 ³Sm、¹ 66 Ho、¹ 77 Lu、¹ 86 Re、¹ 88Radionuclides of Re, At, Pb, and Bi. In a specific embodiment, the radionuclide is ¹ 77 Lu. In a specific implementation scheme, the radionuclide is... 67 Cu.

[0064] In some embodiments, the conjugate has the structure of Formula I or Formula IV: (Formula I) (Form IV); Wherein RN is a radionuclide; where X is 5G or ABM-5G; where 5G is a peptide; and where ABM is the albumin-binding portion.

[0065] In some implementations, X is ABM-5G, such that the resulting therapeutic conjugate is included in Formula II: (Formula II).

[0066] In some implementations, X is 5G, such that the resulting therapeutic conjugate is included in Formula III: (Formula III)

[0067] Such therapeutic and imaging conjugates are not limited to a specific chemical moiety of Ra. In some embodiments, RN is... 67 Cu or 64 Cu. In some embodiments, the RN of the therapeutic conjugate is selected from... 47 Sc、 67 Cu、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 203 Pb, 212 Pb and 225 Ac. In other examples, the RN used for imaging conjugates is selected from... 11 C 13 N、 15 O、 18 F, 55 Co、 60 Cu、 61Cu、 62 Cu、 64 Cu、 66 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 111 In、 124 I, 125 I, 131 I and 203 Pb. In other examples, RN is 177 Lu (therapeutic conjugate) or 68 Ga (imaging conjugate). Such therapeutic conjugates are not limited to 5G as a specific chemical moiety of the peptide. In some embodiments, the peptide is configured to bind α v β6 integrin. In some embodiments, the peptide is configured to bind α. v RGD peptide of β6 integrin. In some embodiments, the peptide is a PEGylated peptide. In some embodiments, the PEGylated peptide has the following amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1) and includes a first PEG moiety at the N-terminus of the peptide and a second PEG moiety at the C-terminus of the peptide. In some embodiments, the first PEG moiety and the second PEG moiety are independently selected from PEG. 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and / or (PEG 28 2 (PEG 1500×2). In some embodiments, the first PEG portion and the second PEG portion are identical. In some embodiments, the first PEG portion and the second PEG portion each contain PEG. 28 (PEG 1500).

[0068] Such conjugates are not limited to a specific chemical moiety of albumin-binding moiety (ABM). In some embodiments, ABM is 4-(4-iodophenyl)butyric acid or K(D-Abu-iodophenylbutyryl) moiety.

[0069] Such therapeutic conjugates are not limited to specific uses or functions. In some embodiments, therapeutic conjugates of formula I, II, III, or IV are used to deliver radioactivity to a patient or subject. In some embodiments, therapeutic conjugates of formula I, II, III, or IV are used to treat or prevent cancer in a patient or subject. In some embodiments, therapeutic conjugates of formula I, II, III, or IV are used to treat alpha-12 cancer. v β6 integrin-related cancers.

[0070] In some embodiments, the conjugate has a structure of formula I, II, III or IV, wherein 5G is a bis-PEGylated RGD peptide.

[0071] In some embodiments, the conjugate has a structure of formula I, II, III, or IV, wherein 5G is a bis-PEGylated RGD peptide, and the RGD peptide has the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1) (e.g., PEG). 28 -GNGVPNLRGDLQVLGQRVGRT-PEG 28 -C(O)NH2).

[0072] In some embodiments, the conjugate has the structure of formula I or III (where X = ABM-5G), and ABM is a 4-(4-iodophenyl)butyric acid moiety or a K(D-Abu-iodophenylbutyryl) moiety.

[0073] In some embodiments, the conjugate comprises a peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1). In some embodiments, the N-terminus and / or C-terminus of the peptide (such as SEQ ID NO: 1) comprises a PEG moiety, such as PEG. 28 (PEG 1500) portion.

[0074] In some embodiments, the conjugate comprises a peptide (such as SEQ ID NO: 1) having a PEG moiety covalently linked to its C-terminus, wherein the PEG moiety is terminated by an amide group, a carboxyl group, or a hydroxyl group.

[0075] In some embodiments, the conjugate comprises a peptide (such as SEQ ID NO: 1) having an N-terminal PEG moiety covalently linked to ABM.

[0076] In some embodiments, the conjugate comprises 1) a peptide (such as SEQ ID NO: 1) having an N-terminal PEG moiety covalently linked to ABM, and 2) a chelating moiety for complexing a radionuclide. In some aspects, the chelating moiety is a DOTA moiety (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) (as covalently linked to the conjugate). In some embodiments, the chelating moiety is a NOA moiety (1,4,7-triazacyclononane-N,N',N”-triacetic acid) (as covalently linked to the conjugate). In some aspects, the chelating moiety of the conjugate does not complex with a radionuclide. In some aspects, the chelating moiety of the conjugate is complexed with a radionuclide (such as...). 11 C 13 N、15 O、 18 F, 47 Sc、 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 89 Sr、 90 Y、 105 Rh、 111 In、 111 Ag、 124 I, 125 I, 117m Sn、 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 203 Pb, 212 Pb and 225 Ac) complexation. In some aspects, the chelating portion of the conjugate is complexed with a radionuclide, and the radionuclide selected for use as a therapeutic agent is, for example... 47 Sc、 67 Cu、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Pb and 212 Bi. In some aspects, the chelated portion of the conjugate is complexed with a radionuclide, and the radionuclide is 177 Lu. In some aspects, the chelated portion of the conjugate is complexed with a radionuclide, and the radionuclide is 67 Cu. In some aspects, the chelated portion of the conjugate is complexed with a radionuclide, and the radionuclide selected for use as a developing agent is, for example... 11 C13 N、 15 O、 18 F, 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 111 In、 124 I, 125 I, 131 I or 203 Pb. In a specific implementation scheme, the radionuclide is... 64 Cu or 67 Cu. In some embodiments, the radionuclide is... 64 Cu, and the conjugate is used for imaging, the radionuclide being... 67 Cu and the conjugate is used for treatment, for example in a therapeutic application (i.e., imaging / diagnosis combined with treatment).

[0077] In some embodiments, the conjugate comprises a peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1) and a complexing agent for radionuclides (e.g., 11 C 13 N、 15 O、 18 F, 32 P, 47 Sc、 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 89 Sr、 90 Y、 105 Rh、 111 In、 111 Ag、 124 I, 125 I, 117m Sn、 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、188 Re、 211 At、 212 Bi、 203 Pb, 212 Pb and 225 The chelating portion of Ac), wherein the N-terminus and C-terminus of the peptide each have a PEG moiety (such as PEG) covalently linked thereto. 28 (PEG 1500) portion). In some embodiments, the PEG portion covalently linked to the C-terminus of the peptide is terminated with an amide, carboxyl, or hydroxyl group. In some aspects, the chelating portion is a DOTA portion (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) covalently linked to the conjugate. In some embodiments, the chelating portion is a NOTA portion (1,4,7-triazacyclononane-N,N',N”-triacetic acid) covalently linked to the conjugate. In some aspects, the chelating portion of the conjugate does not complex with a radionuclide. In some aspects, the chelating portion of the conjugate complexes with a radionuclide. In some aspects, the chelating portion of the conjugate complexes with a radionuclide, and the radionuclide selected for use as a therapeutic agent, for example... 47 Sc、 67 Cu、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Pb, 212 Bihe 225 Ac. In some aspects, the chelated portion of the conjugate is complexed with a radionuclide, and the radionuclide is 177 Lu. In some aspects, the chelated portion of the conjugate is complexed with a radionuclide, and the radionuclide is 67 Cu. In some aspects, the chelated portion of the conjugate is complexed with a radionuclide, and the radionuclide selected for use as a developing agent is, for example... 11 C 13 N、 15 O、 18 F, 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、67 Cu、 68 Ga、 82 Rb、 86 Y、 111 In、 124 I, 125 I, 131 I or 203 Pb. In a specific implementation scheme, the radionuclide is... 67 Cu or 64 Cu. In a specific implementation scheme, the radionuclide is... 67 Cu, and the conjugate is used as a therapeutic agent. In a specific embodiment, the radionuclide is... 64 Cu, and the conjugate is used as an imaging agent. In some embodiments, the conjugate comprises a peptide and a chelating moiety for complexing a radionuclide, the peptide having the amino acid sequence RGDLX1X2X3 (SEQ ID NO: 5), wherein X1 and X2 are independently selected amino acids and X3 is L or I (SEQ ID NO: 2), or the peptide having the amino acid sequence RGDLX1X2X3AQX6 (SEQ ID NO: 6), wherein X6 is K or R, optionally, wherein such a peptide is double-PEGylated (i.e., the N-terminus and C-terminus of the peptide each have a PEG moiety covalently linked thereto, such as PEG). 28 (PEG 1500) portion). In some embodiments, the PEG portion covalently linked to the C-terminus of the peptide is terminated with an amide, carboxyl, or hydroxyl group. In some aspects, the chelating portion is a DOTA portion (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) covalently linked to the conjugate. In some embodiments, the chelating portion is a NOTA portion (1,4,7-triazacyclononane-N,N',N”-triacetic acid) covalently linked to the conjugate. In some aspects, the chelating portion of the conjugate does not complex with a radionuclide. In some aspects, the chelating portion of the conjugate complexes with a radionuclide. In some aspects, the chelating portion of the conjugate complexes with a radionuclide, and the radionuclide selected for use as a therapeutic agent, for example... 47 Sc、 67 Cu、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211At、 212 Pb, 212 Bihe 225 Ac. In some aspects, the chelated portion of the conjugate is complexed with a radionuclide, and the radionuclide is 177 Lu. In some aspects, the chelated portion of the conjugate is complexed with a radionuclide, and the radionuclide is 67 Cu. In some aspects, the chelated portion of the conjugate is complexed with a radionuclide, and the radionuclide selected for use as a developing agent is, for example... 11 C 13 N、 15 O、 18 F, 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 111 In、 124 I, 125 I, 131 I or 203 Pb. In a specific implementation scheme, the radionuclide is... 67 Cu or 64 Cu.

[0078] In some embodiments, any therapeutic conjugate described herein (such as those included in formulas I, II, III, or IV) is provided as a pharmaceutical composition for administration. In some cases, the pharmaceutical composition comprises one or more unit doses, wherein the amount of radioactivity present in the dose is from about 25 mCi to about 200 mCi. In some embodiments, the amount of radioactivity present in a dose of the conjugate is from about 25 mCi to about 50 mCi, from about 25 mCi to about 100 mCi, from about 25 mCi to about 150 mCi, from about 25 mCi to about 200 mCi, from about 50 mCi to about 100 mCi, from about 50 mCi to about 150 mCi, from about 50 mCi to about 200 mCi, from about 100 mCi to about 150 mCi, from about 100 mCi to about 200 mCi, or from about 150 mCi to about 200 mCi. In some cases, the amount of radioactivity present per unit dose is about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi. In some embodiments, the amount of peptide per unit dose of the conjugate does not exceed about 500 μg, about 400 μg, about 300 μg, about 200 μg, or about 100 μg of peptide. In some cases, the amount of peptide per unit dose of the conjugate does not exceed about 100 μg of peptide. In other embodiments, the amount of peptide per unit dose of the conjugate may be greater than about 100 μg of peptide.

[0079] In some embodiments, the conjugate is a diagnostic conjugate and comprises a diagnostic agent. In some embodiments, the diagnostic agent is a radionuclide, such as a positron-emitting radionuclide. In some embodiments, the diagnostic conjugate herein comprises a radionuclide, such as... 11 C 13 N、 15 O、 18 F, 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Cu、 68 Ga、 203 Pb, 82 Rb、 86 Y、 111 In、 124 I, 125 I or 131 I. In the specific implementation plan, the radioactive nuclide is... 64Cu. In some embodiments, the conjugate has the structure of formula I, II, III, or IV, wherein 5G is a bis-PEGylated RGD peptide comprising the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1). In some further preferred embodiments, X is ABM-5G, the 5G peptide is SEQ ID NO: 1, and ABM is 4-(4-iodophenyl)butyric acid or K(D-Abu-iodophenylbutyryl) moiety, and RN is most preferably... 64 Cu or 67 Cu.

[0080] IV. Instructions for Use

[0081] The method described herein involves providing a therapeutically effective dose of a therapeutic conjugate, as described herein, to a subject. The therapeutic conjugate described herein may contain a radionuclide, such as... 67 Cu. In some embodiments, the conjugate has formula I ( ) or formula IV ( The structure of ) is preferred, where RN is a radioactive nuclide. 67 Cu, where X is ABM-5G or 5G.

[0082] In some implementations, X is ABM-5G, such that the resulting therapeutic conjugate is included in Formula II: (Formula II).

[0083] In some implementations, X is 5G, such that the resulting therapeutic conjugate is included in Formula III: (Formula III)

[0084] In some embodiments, 5G is a double-PEGylated RGD peptide. In some embodiments, the RGD peptide (i.e., present in conjugates having the structure of formula I, II, III, or IV) has the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), for example, PEG. 28 -GNGVPNLRGDLQVLGQRVGRT-PEG 28-C(O)NH2. In some embodiments, the RGD peptide (i.e., present in conjugates having the structure of formula I, II, III, or IV) has the amino acid sequence RGDLX1X2X3 (SEQ ID NO: 5), wherein X1 and X2 are independently selected amino acids and X3 is L or I (SEQ ID NO: 2); or has the amino acid sequence RGDLX1X2X3AQX6 (SEQ ID NO: 6), wherein X6 is K or R, optionally wherein such a peptide is double-PEGylated. In some further preferred embodiments, X is AMB-5G, ABM is the albumin-binding moiety (such as 4-(4-iodophenyl)butyric acid or K(D-Abu-iodophenylbutyryl) moiety), and RN is a radionuclide.

[0085] In some embodiments, the dose of the therapeutic conjugate administered to the subject contains about 25 mCi to about 200 mCi of radioactivity. In some embodiments, the dose of the conjugate contains about 25 mCi to about 100 mCi of radioactivity. In some embodiments, the dose of the therapeutic conjugate contains about 25 mCi to about 150 mCi of radioactivity. In some cases, the administered dose contains about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi of radioactivity. In some embodiments, the dose of the therapeutic conjugate does not cause adverse events (AEs) in the subject, such as grade 3 AEs (i.e., serious AEs). In some embodiments, the dose (or cumulative dose) of the therapeutic conjugate does not exceed a radiation dose of about 23 Gy to the kidneys and / or about 1.5 Gy to the bone marrow.

[0086] The therapeutic conjugate can be administered by infusion, for example over a period of time, such as several minutes or several hours. In some embodiments, the therapeutic conjugate is infused over several minutes, such as about 1 minute, about 2 minutes, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, or about 55 minutes. In some embodiments, the therapeutic conjugate is infused over about 30 minutes. In some embodiments, the therapeutic conjugate is infused over several hours, such as about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, or about 8 hours. In some embodiments, the therapeutic conjugate is infused over about 4 hours. In some embodiments, the therapeutic conjugate is co-infused with an amino acid solution. In some cases, the amino acid solution is infused before the therapeutic conjugate is infused.

[0087] In some implementations, the therapeutic conjugate is administered to the subject once, twice, three times, four times, or five times during treatment. Subsequent administrations of the therapeutic conjugate may be performed at predetermined time intervals (days, weeks, or months). In some cases, if a tumor or cancer cells reappear, continue to grow, or are not fully treated after the first administration of the therapeutic conjugate, the therapeutic conjugate is administered at a subsequent time. In some cases, if the subject does not fully respond to the first treatment, experiences a partial response, a stable response, or progressive disease, the therapeutic conjugate is administered again at a subsequent time.

[0088] In some embodiments, the dosimetry and / or biodistribution of the therapeutic conjugate are evaluated after administration to the subject. As a non-limiting example, the dosimetry and biodistribution of the therapeutic conjugate may be evaluated using radionuclide imaging approximately 1 day and / or approximately 7 days (e.g., 5 days, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 21 days, 28 days) after administration (e.g., infusion) to the subject. In some cases, approximately 24 hours and / or approximately 168 hours after administration of the therapeutic conjugate, the subject undergoes whole-body planar imaging (e.g., anterior and posterior views) and single-photon emission computed tomography / computed tomography (SPECT / CT) (e.g., extending from the top of the skull to the perineum, ending at the proximal thigh; approximately 2-4 beds). In some cases, serial blood samples are drawn approximately 5, 15, 30, 60, 120, and / or 180 minutes after administration of the therapeutic conjugate, for example, to evaluate biodistribution. In some cases, full chemistry, hematology, liver function tests, and / or EKG are performed approximately 1 day and / or approximately 7 days (e.g., ±48 hours) after administration of the therapeutic conjugate.

[0089] Dosimetry analysis methods are known in the art and include, but are not limited to: descriptive statistics (e.g., mean, median, standard deviation, etc.) of AUC reported based on the activity concentration-time curve of the therapeutic conjugate (e.g., for identifiable thoracic and abdominal organs, target lesions, and blood), maximum uptake at target lesions and identifiable organs (e.g., obtained as a percentage), specific absorbed dose (μGy / MBq) for each organ, and cumulative absorbed dose (Gy) for each organ. In some cases, the frequency and percentage of organs receiving the highest absorbed dose are tabulated by tissue equivalent dose assessment. In some cases, graphical tools are used to depict endpoints.

[0090] In some implementations, the distribution of the therapeutic conjugate is determined using whole-body planar SPECT / CT imaging. As a non-limiting example, the absorbed radiation dose to the kidneys, stomach, unaffected liver, bone marrow, and any other organs showing accumulation of the therapeutic conjugate is calculated based on analysis of consecutive blood counts and SPECT / CT scans. In some cases, SPECT / CT images are used to calculate the volumetric absorbed radiation dose in diseased and healthy tissues; for example, the activity concentration-time curve for normal tissue can be generated from region of interest (ROI) analysis of the SPECT / CT scan, the activity concentration-time curve for red bone marrow and heart can be generated from blood activity concentrations measured by a well-type scintillation counter, and / or the volume of interest (VOI) for each patient can be generated. In some cases, the activity concentration in red bone marrow is equal to the activity concentration in blood. In some cases, the activity concentration-time curve is integrated (e.g., using analytical or numerical methods as appropriate) to obtain an AUC value, from which the so-called residence time is derived. In some cases, these data are input into organ dosimetry software (such as OLIDA / EXM) to generate an estimate of the absorbed dose to normal tissue. In some cases, supplemental dosimetry assessments are performed, including, for example, lesion absorption dose estimation based on image ROI analysis. In some cases, the absorbed dose is normalized to the administered activity and expressed in mGy / MBq.

[0091] In some embodiments, the distribution of the therapeutic conjugate in the tumor (e.g., primary tumor or cancerous lesion), blood, gallbladder, liver, heart, lungs, spleen, kidneys, pancreas, stomach, small intestine, bladder, skin, muscle, bone, large intestine, and / or brain of the target is determined using, for example, SPECT / CT imaging. In some embodiments, the amount of the therapeutic conjugate present in non-tumor tissue approximately 24 hours, approximately 48 hours, and / or approximately 72 hours after application of the conjugate is lower than the amount present in non-tumor tissue approximately 1 hour after application of the conjugate. In some embodiments, the ratio of the amount of the therapeutic conjugate in the tumor to the amount of the therapeutic conjugate in non-tumor tissue approximately 24 hours, approximately 48 hours, and / or approximately 72 hours after application of the conjugate is higher than the ratio of the amount of the therapeutic conjugate in the tumor to the amount of the therapeutic conjugate in non-tumor tissue approximately 1 hour after application of the conjugate.

[0092] In some embodiments, the amount of the therapeutic conjugate present in kidney tissue approximately 24 hours, approximately 48 hours, and / or approximately 72 hours after application of the conjugate is lower than the amount of the therapeutic conjugate present in kidney tissue 1 hour after application of the conjugate. In some embodiments, the ratio of the amount of the therapeutic conjugate in the primary tumor to the amount of the therapeutic conjugate in kidney tissue approximately 24 hours, approximately 48 hours, and / or approximately 72 hours after application of the conjugate is higher than the ratio of the amount of the therapeutic conjugate in the primary tumor to the amount of the therapeutic conjugate in kidney tissue 1 hour after application of the conjugate.

[0093] In some embodiments, the amount of the therapeutic conjugate present in gastric tissue approximately 24 hours, approximately 48 hours, and / or approximately 72 hours after application of the conjugate is lower than the amount of the therapeutic conjugate present in gastric tissue approximately 1 hour after application of the conjugate. In some embodiments, the ratio of the amount of the therapeutic conjugate in the primary tumor to the amount of the therapeutic conjugate in gastric tissue approximately 24 hours, approximately 48 hours, and / or approximately 72 hours after application of the conjugate is higher than the ratio of the amount of the therapeutic conjugate in the primary tumor to the amount of the therapeutic conjugate in gastric tissue approximately 1 hour after application of the conjugate.

[0094] In some embodiments, the amount of the therapeutic conjugate present in the colonic tissue approximately 24 hours, approximately 48 hours, and / or approximately 72 hours after application of the conjugate is lower than the amount of the therapeutic conjugate present in the colonic tissue approximately 1 hour after application of the conjugate. In some embodiments, the ratio of the amount of the therapeutic conjugate in the primary tumor to the amount of the therapeutic conjugate in the colonic tissue approximately 24 hours, approximately 48 hours, and / or approximately 72 hours after application of the conjugate is higher than the ratio of the amount of the therapeutic conjugate in the primary tumor to the amount of the therapeutic conjugate in the colonic tissue approximately 1 hour after application of the conjugate.

[0095] In some embodiments, the amount of the therapeutic conjugate in liver tissue is minimal when measured approximately 1 hour, approximately 24 hours, approximately 48 hours, and / or approximately 72 hours after administration of the conjugate. In some embodiments, the amount of the therapeutic conjugate in primary tumors is significantly greater than the amount in liver tissue at approximately 1 hour, approximately 24 hours, approximately 48 hours, and / or approximately 72 hours after administration of the conjugate.

[0096] In some embodiments of the method described herein, a diagnostic conjugate is administered to the subject prior to the administration of the therapeutic conjugate. The diagnostic conjugate comprises an RGD peptide that binds to an α-type radionuclide covalently linked (directly or indirectly) to a radionuclide. vβ6 integrin. In some cases, the RGD peptide of the diagnostic conjugate has the same amino acid sequence as the RGD peptide present in the therapeutic conjugate. In some embodiments, the radionuclide of the diagnostic conjugate may have a shorter half-life compared to the radionuclide of the therapeutic conjugate. In some cases, the radionuclide is... 64 Cu. In some cases, the radionuclide used to diagnose conjugates is... 64 Cu, and the therapeutic conjugate contains a radionuclide. 67 Cu. In some embodiments, the diagnostic conjugate has the structure of Formula I or Formula IV: (Formula I) (Formula IV) Where RN is a radioactive nuclide (preferred). 64 Cu), where X is ABM-5G or 5G.

[0097] In some implementations, X is ABM-5G, such that the resulting therapeutic conjugate is included in Formula II: (Formula II).

[0098] In some implementations, X is 5G, such that the resulting therapeutic conjugate is included in Formula III: (Formula III)

[0099] In some embodiments, the diagnostic conjugate has the structure of Formula I, wherein 5G is a bis-PEGylated RGD peptide. In some embodiments, the RGD peptide (i.e., present in diagnostic conjugates having the structure of Formula I, II, III, or IV) has the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), for example, PEG. 28 -GNGVPNLRGDLQVLGQRVGRT-PEG 28 -C(O)NH2. In some further preferred embodiments, X is ABM-5G, and AMB is an albumin-binding moiety (such as 4-(4-iodophenyl)butyric acid or K(D-Abu-iodophenylbutyryl) moiety).

[0100] In some embodiments, the method includes administering a diagnostic conjugate of formula I, II, III, or IV for diagnostic imaging of tumors, cancerous lesions, or cancer cells. In some embodiments, the method includes administering a diagnostic conjugate of formula I, II, III, or IV for diagnostic imaging of tumors, cancerous lesions, or cancer cells, and subsequently administering a therapeutic conjugate of formula I, II, III, or IV to treat the tumor, cancerous lesion, or cancer cells. In some embodiments, the method includes administering a therapeutic conjugate of formula I, II, III, or IV to treat the cancerous lesion when sufficient lesion uptake of the diagnostic conjugate of formula I, II, III, or IV is detected (e.g., any visualized lesion with a maximum normalized uptake value (SUVmax) greater than twice that of normal lung or liver).

[0101] In some embodiments, the diagnostic conjugate is administered by injection. In some embodiments, the diagnostic conjugate is administered by infusion. In some embodiments, the diagnostic conjugate is infused over a period of several minutes, such as about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, or less. In some embodiments, the diagnostic conjugate is infused over a period of about 5 minutes or less.

[0102] In some embodiments, the diagnostic conjugate can be visualized in the body or a part of the body of the subject. In some embodiments, absorption and accumulation of the diagnostic conjugate in a lesion, tissue, or organ can be used to select subjects for treatment with the therapeutic conjugate. In some embodiments, the diagnostic conjugate is used to select subjects with α v Patients with β6 integrin-related lesions or cancer cells who are suitable for treatment with the therapeutic conjugate. In some embodiments, the diagnostic conjugate is used to select patients with α v Patients with β6 integrin-related lesions or cancer cells are selected, and then the selected patients receive a dose of the therapeutic conjugate, such as a therapeutic conjugate of formula I, II, III, or IV, within 1, 2, 3, 4, 5, or 6 weeks after imaging with the diagnostic conjugate. In some embodiments, the diagnostic conjugate is used to select patients with α v Patients with β6 integrin-related lesions or cancer cells, and then selected patients receive a dose of a therapeutic conjugate, such as a therapeutic conjugate of formula I, II, III or IV, within 4 to 5 weeks after imaging the diagnostic conjugate.

[0103] In some embodiments, the overall distribution of the diagnostic conjugate is determined through visual analysis of blood data and PET / CT scans. As a non-limiting example, reconstructed PET / CT images (such as a whole-body static imaging from the top of the head to the proximal thigh) can be displayed on an imaging workstation and recalibrated into maximum intensity projection (MIP), axial, coronal, and sagittal images. In some embodiments, PET, fused PET / CT, and / or CT images are evaluated. In some embodiments, the region of interest (ROI) is placed around the tracer focus of suspected malignant tumors and key organs (such as the kidneys, bladder, intestines, liver, spleen, lungs, and pancreas) to obtain SUV parameters, including SUVmax and SUV mean. In some cases, SUV measurements are summarized using the mean, median, numerical range, and count, and the SUV is correlated with the tissue region using a repeated measures ANOVA model. In some embodiments, “unexcreted” and “excreted” radioactivity are tracked, where unexcreted radioactivity in the body is calculated by volume of interest (VOI) analysis, thereby obtaining the amount of radioactivity in major organs, tissues of interest, and the remainder of the body. In some cases, for dosimetry purposes, data are converted into a "reference person" anthropomorphic model. In other cases, organ activity is integrated over time to obtain the time-integrated activity coefficient. In still other cases, organ dosimetry software (such as OLINDA / EXM) is used to obtain dose and effective dose measurements.

[0104] Any apparatus or method known in the art for detecting radioactive emission of a radionuclide in a subject is suitable for use with the conjugates and methods described herein. For example, methods such as single-photon emission computed tomography (SPECT) and radionuclide scintillation can be used to detect radiation emitted from the radiolabeled conjugates described herein, wherein SPECT uses a rotating gamma camera to detect radiation from a single-photon gamma-emitting radionuclide, and radionuclide scintillation uses a scintillation gamma camera to obtain images or a series of sequential images of the distribution of the radionuclide in a tissue, organ, or body system. Positron emission tomography (PET) is another suitable technique for detecting radiation in a subject. Furthermore, U.S. Patent No. 5,429,133 describes a laparoscopic probe for detecting radiation concentrated in a tumor in solid tissue. Miniature and flexible radiation detectors intended for medical use are manufactured by Intra-Medical LLC (Santa Monica, CA). Furthermore, nuclear magnetic resonance (NMR) based methods (such as magnetic resonance spectroscopy (MRS) and magnetic resonance imaging (MRI)) or any other imaging technique known to those skilled in the art (including, but not limited to, computed tomography (CT)) can be combined with methods suitable for detecting the radioactive emission of radionuclides. In some embodiments, radiation from a radionuclide is used to determine where a conjugate (such as a diagnostic conjugate as described herein) is concentrated in the subject. Regardless of the method or apparatus used, such detection aims to determine where the conjugate is concentrated in the subject, and such concentration is an indicator of the location of a tumor or tumor cells.

[0105] In some implementation schemes, patients with cancer (such as alpha) vSubjects of β6 integrin-associated cancers may be given the therapeutic conjugates described herein (such as therapeutic conjugates of formulas I, II, III, or IV). Non-limiting examples of different types of cancer suitable for treatment with the therapeutic conjugates described herein include lung cancer, breast cancer, pancreatic cancer, bladder cancer, thyroid cancer, liver cancer, pleural cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, testicular cancer, colon cancer, colorectal cancer, anal cancer, bile duct cancer, gastrointestinal carcinoid tumors, esophageal cancer, gallbladder cancer, rectal cancer, appendix cancer, small bowel cancer, stomach cancer, kidney cancer (i.e., renal cell carcinoma), central nervous system cancers, skin cancer, and oral squamous cell carcinoma. The cancers include squamous cell carcinoma of the skin, choriocarcinoma, head and neck cancer, bone cancer, osteosarcoma, fibrosarcoma, neuroblastoma, glioma, melanoma, leukemia (such as acute lymphoblastic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, or hairy cell leukemia), lymphoma (such as non-Hodgkin's lymphoma, Hodgkin's lymphoma, B-cell lymphoma, or Burkitt's lymphoma), and multiple myeloma. In some embodiments, the cancer is lung cancer, breast cancer, colorectal cancer, pancreatic cancer, ovarian cancer, cervical cancer, oral squamous cell carcinoma, squamous cell carcinoma of the skin, gastric cancer, or endometrial cancer. In some cases, the subject has a primary lesion (such as a primary tumor). In some cases, the subject has metastases (such as metastatic forms of any cancer type described herein). In some cases, the subject has both a primary lesion and metastases. In some implementations, the subject has pancreatic cancer, such as locally advanced or metastatic pancreatic cancer, locally advanced, unresectable or metastatic pancreatic adenocarcinoma or pancreatic ductal adenocarcinoma (PDAC).

[0106] In some implementations, the subject receives a dose of a therapeutic conjugate of formula I, II, III, or IV containing about 25 mCi to about 200 mCi of radioactivity, such as a dose having about 25 mCi, about 50 mCi, about 100 mCi, about 15 mCi, or about 20 mCi of radioactivity, administered over several minutes or hours (e.g., about 1 minute, about 2 minutes, about 5 minutes, about 10 minutes, about 150 mCi, or about 200 mCi of radioactivity). In some embodiments, the subject receives a dose of a therapeutic conjugate of formula I, II, III, or IV containing about 25 mCi to about 200 mCi of radioactivity, administered over a period of about 30 minutes. For example, the dose may have about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi of radioactivity.

[0107] In some embodiments, the subject receives standard care treatment prior to administration of the therapeutic conjugate dose. In some aspects, the therapeutic conjugate has a structure of formula I, II, III, or IV, and the subject receives a dose of the therapeutic conjugate containing about 25 mCi to about 200 mCi of radioactivity, for example, a dose having about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi of radioactivity, and prior to such a dose, the subject receives standard care treatment. Standard care treatment may include one or more of surgery, radiotherapy, chemotherapy, chemoradiotherapy, and targeted therapy. Standard care treatment may include FOLFIRINOX (leucovorin calcium (leucovorin), fluorouracil, irinotecan hydrochloride, and oxaliplatin). Standard care treatment may include gemcitabine, albumin-bound paclitaxel, or a combination thereof. Standard care treatment may include irinotecan. Standard care treatment may include surgery or surgery and one or more of FOLFIRINOX, gemcitabine, albumin-bound paclitaxel, and irinotecan.

[0108] In some embodiments, the subject receives standard care treatment after administration of the therapeutic conjugate at the dose stated therein. In some aspects, the therapeutic conjugate has a structure of formula I, II, III, or IV, and the subject receives a dose of the therapeutic conjugate containing about 25 mCi to about 200 mCi of radioactivity, for example, a dose having about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi of radioactivity, and after such a dose, the subject receives standard care treatment. Standard care treatment may include one or more of surgery, radiotherapy, chemotherapy, chemoradiotherapy, and targeted therapy. Standard care treatment may include FOLFIRINOX (leucovorin calcium (leucovorin), fluorouracil, irinotecan hydrochloride, and oxaliplatin). Standard care treatment may include gemcitabine, albumin-bound paclitaxel, or a combination thereof. Standard care treatment may include irinotecan. Standard care treatment may be surgery or surgery and one or more of FOLFIRINOX, gemcitabine, albumin-bound paclitaxel, and irinotecan.

[0109] In some embodiments, the subject receives one or more standard care treatments before and after administration of the therapeutic conjugate dose. In some aspects, the therapeutic conjugate has a structure of formula I, II, III, or IV, and the subject receives a dose of the therapeutic conjugate containing about 25 mCi to about 200 mCi of radioactivity, for example, a dose having about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi of radioactivity, and receives standard care treatment before and after such a dose. Standard care treatment may include one or more of surgery, radiotherapy, chemotherapy, chemoradiotherapy, and targeted therapy. Standard care treatment may include FOLFIRINOX (leucovorin calcium (leucovorin), fluorouracil, irinotecan hydrochloride, and oxaliplatin). Standard care treatment may include gemcitabine, albumin-bound paclitaxel, or a combination thereof. Standard care treatment may include irinotecan. Standard care may include surgery or surgical procedures and one or more of FOLFIRINOX, gemcitabine, albumin-bound paclitaxel, and irinotecan. Standard care may be the same before and after treatment with the therapeutic conjugate. Standard care may also differ between the two.

[0110] In some embodiments, treatment with the therapeutic conjugate results in stable, partial, or complete remission of the subject's condition (e.g., the methods described herein include administering a dose of the therapeutic conjugate to the subject that kills or otherwise slows the growth or progression of cancer cells and results in stable or partial or complete remission of the cancer). In some embodiments, treatment with the therapeutic conjugate results in a reduction of cancer metastasis in the subject (e.g., the methods described herein include administering a dose of the therapeutic conjugate to the subject that reduces cancer metastasis in the subject). In some embodiments, treatment with the therapeutic conjugate results in a reduction of the volume, size, or growth of a tumor in the subject (e.g., the methods described herein include administering a dose of the therapeutic conjugate to the subject that reduces the volume, size, or growth of a tumor in the subject). In some embodiments, treatment with the therapeutic conjugate results in an increased response of the cancer to a subsequently administered anticancer agent (e.g., the methods described herein include administering a dose of the therapeutic conjugate to the subject that increases the response of the cancer to a subsequently administered anticancer agent).

[0111] In some embodiments, a subject is treated with a therapeutic conjugate of formula I, II, III, or IV, and the subject receives a dose of the therapeutic conjugate containing about 25 mCi to about 200 mCi of radioactivity, such as a dose having about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi of radioactivity, and treatment with the therapeutic conjugate results in stable, partial, or complete remission of the subject's condition (e.g., the dose of the therapeutic conjugate kills or otherwise slows the growth or progression of cancer cells and results in stable or partial or complete remission of cancer in the subject).

[0112] In some embodiments, a subject is treated with a therapeutic conjugate of formula I, II, III, or IV, and the subject receives a dose of the therapeutic conjugate containing about 25 mCi to about 200 mCi of radioactivity, for example, a dose having about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi of radioactivity, and treatment with the therapeutic conjugate results in a reduction in cancer metastasis in the subject (e.g., the dose of the therapeutic conjugate reduces cancer metastasis in the subject).

[0113] In some embodiments, a subject is treated with a therapeutic conjugate of formula I, II, III, or IV, and the subject receives a dose of the therapeutic conjugate containing about 25 mCi to about 200 mCi of radioactivity, for example, a dose having about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi of radioactivity, and treatment with the therapeutic conjugate results in a reduction in the volume, size, or growth of a tumor in the subject (e.g., the dose of the therapeutic conjugate reduces the volume, size, or growth of a tumor in the subject).

[0114] In some embodiments, a subject is treated with a therapeutic conjugate of formula I, II, III, or IV, and the subject receives a therapeutic conjugate containing a dose of radioactivity of about 25 mCi to about 200 mCi, for example, a dose having radioactivity of about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi, and treatment with the therapeutic conjugate results in an increased response of the cancer to a subsequently administered anticancer agent (e.g., the dose of the therapeutic conjugate increases the response of the cancer to a subsequently administered anticancer agent).

[0115] In some embodiments, the subject has pancreatic cancer, such as locally advanced, unresectable, or metastatic pancreatic adenocarcinoma, and is treated with a therapeutic conjugate of formula I, II, III, or IV. The subject receives a dose of the therapeutic conjugate containing about 25 mCi to about 200 mCi of radioactivity, for example, a dose having about 25 mCi, about 50 mCi, about 100 mCi, about 150 mCi, or about 200 mCi of radioactivity. In some aspects, the treatment results in stable, partial, or complete remission of the subject's condition (e.g., the dose of the therapeutic conjugate kills or otherwise slows the growth or progression of cancer cells, resulting in stable or partial or complete remission of the cancer). In some aspects, the treatment results in a reduction of pancreatic cancer metastasis in the subject (e.g., the dose of the therapeutic conjugate reduces cancer metastasis in the subject). In some aspects, the treatment results in a reduction in the volume, size, or growth of the pancreatic tumor in the subject (e.g., the dose of the therapeutic conjugate reduces the volume, size, or growth of the tumor in the subject). In some respects, treatment leads to an increased response of the cancer to subsequently administered anticancer agents (such as chemotherapy agents or combinations thereof) (e.g., the therapeutic conjugate at said dose increases the cancer's response to subsequently administered anticancer agents). In some respects, treatment leads to an increased response of the cancer to subsequently administered anticancer agents (such as FOLFIRINOX, gemcitabine, albumin-bound paclitaxel, irinotecan, or any combination thereof).

[0116] In a further embodiment, the present invention provides a treatment for α v A method for treating β6 integrin-associated cancer, comprising administering a dose of a therapeutic conjugate to a subject, wherein the therapeutic conjugate comprises a peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), a chelating moiety, and a radionuclide, wherein the peptide further comprises a first PEG moiety at the N-terminus of the peptide and a second PEG moiety at the C-terminus of the peptide, wherein the radionuclide is preferably... 67 Cu.

[0117] In some embodiments, the therapeutic conjugate comprises a chelating moiety. The invention is not limited to any particular chelating moiety. In some preferred embodiments, the chelating moiety is NOA as described above. In other embodiments, the chelating moiety is DOTA. Preferred DOTA conjugates are described in WO2022120226, which is incorporated herein by reference in its entirety. In some embodiments, the conjugate further comprises an albumin-binding moiety (ABM). Preferred ABMs are as described above. For example, a DOTA conjugate includes, for example, the following formula: DOTA-ABM-5G; (Form IV), ABM-5G and RN are as described in this article.

[0118] In some preferred embodiments, the treatment method comprises administering more than one dose of the therapeutic conjugate to the subject. In some particularly preferred embodiments, two, three, or four doses (or more) are administered during treatment. In some embodiments, the amount of the therapeutic conjugate in the subject's kidney is reduced by at least two-fold within 24 hours following the administration of the dose of the therapeutic conjugate. In some aspects of such preferred embodiments, the therapeutic conjugate has a structure of any one of formula I, II, III, or IV, and the radionuclide is... 67 Cu. In some particularly preferred embodiments, two, three, or four doses (or more) are administered during treatment, the conjugate having the structure of Formula II, and the radionuclide being Cu. 67 In some particularly preferred embodiments, two, three, or four doses (or more) are administered during treatment, the conjugate having the structure of Formula I, and the radionuclide being... 67 Cu. In some particularly preferred embodiments, two, three, or four doses (or more) are administered during treatment, the conjugate having the structure of formula IV, and the radionuclide being... 67 Cu.

[0119] V. Pharmaceutical Composition

[0120] The therapeutic and diagnostic conjugates described herein can be formulated into pharmaceutical compositions for use. Such pharmaceutical compositions may comprise the conjugate and one or more pharmaceutically acceptable excipients suitable for injection and / or infusion.

[0121] The conjugates described herein can be formulated into compositions for administration in liquid form. The liquid can be delivered by injection, such as intratumoral injection or intravenous infusion. The compositions for administration by injection may also include one or more of surfactants, preservatives, wetting agents, dispersants, suspending agents, buffers, stabilizers (such as radiation protectants), and isotonic agents.

[0122] Liquid compositions (whether solutions, suspensions, or other similar forms) may also include one or more of the following: sterile diluents (such as water for injection), saline solutions (preferably physiological saline), Ringer's solutions, isotonic sodium chloride, non-volatile oils (such as synthetic monoglycerides or diglycerides that can be used as solvents or suspension media), polyethylene glycol, glycerol, cyclodextrin, propylene glycol, or other solvents; antimicrobial agents, such as benzyl alcohol or methylparaben; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as amino acids, acetates, citrates, or phosphates; detergents, such as nonionic surfactants, polyols; and agents for adjusting tension, such as sodium chloride or dextrose. Liquid compositions may include aqueous or oily suspensions or emulsions, as well as sesame oil, corn oil, cottonseed oil, or peanut oil, and elixirs, mannitol, dextrose, or sterile aqueous solutions, and similar pharmaceutical mediators. Aqueous solutions of saline are also routinely used for injection. Ethanol, glycerol, propylene glycol and liquid polyethylene glycol (and suitable mixtures thereof), cyclodextrin derivatives and vegetable oils can also be used. Appropriate flowability can be maintained, for example, by using coatings (such as lecithin) to maintain the desired particle size in the dispersed state, and by using surfactants to maintain appropriate flowability.

[0123] The compositions used herein are preferably sterile. Microbial activity can be prevented by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, and thimerosal.

[0124] Compositions for administering the conjugates herein typically contain at least one excipient, such as water, physiological saline (0.9% NaCl), or cell buffer for infusion, preferably composed of physiological saline solutions replaced with a protein component such as human serum albumin (HAS).

[0125] The conjugate composition for application can be encapsulated in ampoules, disposable syringes, or multi-dose vials made of glass, plastic, or other materials.

[0126] In some cases, the diagnostic conjugate is prepared in 0.9% sodium chloride solution (USP), wherein anhydrous ethanol ≤10% v / v. In some cases, the therapeutic conjugate is prepared in 0.9% sodium chloride solution (USP) at a concentration of 5 mg / mL sodium ascorbate (USP), wherein anhydrous ethanol ≤10% v / v.

[0127] VI. Examples

[0128] The following embodiments are provided for illustration and not for limitation of the subject matter for which protection is claimed.

[0129] Example 1: Therapeutic Agent / Diagnostic Agent

[0130] method

[0131] Peptide 1 (DOTA-ABM-5G) was synthesized using the standard Fomc solid-phase chemical method. );and

[0132] Peptide 2 (NOTA-ABM-5G; ). Assess α using ELISA v The affinity of β6. The peptide was tested at 37°C with copper-64 in 0.1 M sodium acetate buffer (pH = 6). 64 Cu) or copper-67 ( 67 Cu was radiolabeled for 15 minutes. In α... v In vitro cell binding and internalization were evaluated in β6(+) human pancreatic cancer Capan-1 cells, and serum stability was assessed in human serum. The study was conducted in female nu / nu mice bearing subcutaneous Capan-1 tumors. 64 Cu-1 and 64 PET / CT imaging and biodistribution of Cu-2 (4 hours, 24 hours, 48 ​​hours, 72 hours).

[0133] result

[0134] In ELISA, peptide 1 and peptide 2 showed effects on α. v The high affinity of β6 (8.4±0.8 and 10.5±1 nM, respectively). Both peptides were used... 64 Cu and 67 Cu was successfully radiolabeled with an activity of 0.5 Ci / μmol molar and a radiochemical purity (RCP) > 98%. This was observed in Capan-1 cells. 64 Cu-1, 64 Cu-2, 67 Cu-1 and 67 Cu-2 exhibits rapid in vitro binding (18-35% binding in 1 hour) and internalization (>50% binding in 1 hour). Figure 1 A), and all peptides showed >95% stability in human serum after 48 hours.

[0135] Capan-1 tumors have 64 Cu-1 and 64 Cu-2 is clearly visible in PET / CT images. Figure 2 Biodistribution studies confirmed the %ID / g at 4 hours and 72 hours over time, representing uptake from the tumor and slow elution from it. 64 Cu-1 was 4.3±0.9 and 2.4±0.2; 64Cu-2 was 4.2 ± 0.3 and 2.03 ± 0.2 (Figure 3). Both peptides showed clearance from the stomach over time, %ID / g at 4 hours and 72 hours: 64 Cu-1 was 12.4±1.7 and 2.9±0.3; 64 Cu⁻² was 12.9 ± 1.2 and 1.5 ± 0.0 (Figure 3). 64 Renal uptake of Cu-2 is significantly lower than 64 Cu-1 (2.4±0.3 vs. 5.8±1.0; P = 0.006) resulted in a tumor-to-kidney ratio improvement of 0.85±0.1 at 72 h. 64 Cu-2) and 0.43±0.0 ( 64 Cu-1). and 64 Compared to Cu-1, 64 Cu-2 also showed partial hepatobiliary clearance, with %ID / g at 24 and 72 hours: 4.6±0.7 and 2.6±0.4; 1.5±0.2 and 1.4±0.2. 64 Cu-1 and 64 Tumor uptake of Cu-2 and current radiotherapy peptides 177 Lu-1 is equivalent to ( Figure 4A ),and 64 Cu-1 and 64 Renal clearance of Cu-2 was improved in all cases; % ID / g at 72 hours: 10.6 ± 2.7 ( 177 Lu-1), 5.8±1 ( 64 Cu-1) and 2.4±0.3 ( 64 Cu-2)(Figure 4).

[0136] in conclusion

[0137] Peptide 1 and peptide 2 against integrin α v β6 has high affinity, and radiolabeled peptides were synthesized using high RCP. 64 Cu-1, 64 Cu-2, 67 Cu-1 and 67 Cu-2. and 177 Compared to Lu-DOTA-ABM-5G, 64 Cu-1 and 64 Cu-2 showed a significant decrease in renal uptake at a later time point, while in mice α v This uptake is maintained in β6(+) tumors. Because 64 Cu / 67 Cu is a "true therapeutic" radionuclide pair, therefore the imaging peptide ( 64 Pharmacokinetics of Cu and therapeutic analogues67 The pharmacokinetics of Cu were matched. Although the peptide chelator structures modified from DOTA(1) to NOA(2) significantly reduced renal uptake, increased hepatobiliary clearance was also observed.

[0138] Example 2: Multiple doses administered

[0139] Based on the kidney's response to... 64 Lower exposure of Cu peptides (see Figure 4), with therapeutic peptides 64 Cu-1 was used in a multi-dose experiment. BxPC-3 tumor cells were subcutaneously implanted into one side of 6-8 week old female mice. The tumors were allowed to grow to a size of 15-50 mm² after implantation for approximately 19 days. 3 (Day 0). On Day 0, mice were divided into 3 groups. Group 1 was the control group treated with saline. Group 2 received a single dose of 37 MBq on Day 0. 64 Cu-1. Group 3 received four doses on days 0, 7, 14, and 28, each dose being 37 MBq. 67 Cu-1. For example... Figure 5 As shown, the mean tumor volume in the animals was assessed from day 0 to day 42. Compared with groups 1 and 2, the 4-dose group (group 3) showed a significant reduction in tumor growth. These results indicate that... 67 Cu-peptide therapy is effective, and its efficacy can be enhanced with multiple doses. Given its relationship with... 177 Compared to Lu-1 64 Cu-1 unexpectedly reduced kidney exposure, and these results suggest that selecting copper as a radioligand for therapeutic (and diagnostic / imaging) purposes offers unexpected benefits in terms of the ability to treat tumors with multiple doses and reduce kidney exposure.

[0140] Exemplary Implementation

[0141] Exemplary embodiments provided based on the currently disclosed subject matter include, but are not limited to, the claims and the following embodiments: 1. Treatment of α v A method for treating β6 integrin-related cancers, comprising administering a dose of a therapeutic conjugate of formula I or IV to a subject requiring treatment: (Formula I) (Formula IV) Wherein RN is a radionuclide, where X is 5G or ABM-5G, where 5G is a PEGylated peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1) and comprising a first PEG portion at the N-terminus of the peptide and a second PEG portion at the C-terminus of the peptide, and where ABM is an albumin-binding portion; If X is ABM-5G, then the resulting therapeutic conjugate is included in Formula II: (Formula II); If X is 5G, then the resulting therapeutic conjugate is included in Formula III: (Formula III)

[0142] 2. The method as described in embodiment 1, wherein the radionuclide is selected from: 11 C 13 N、 15 O、 18 F, 32 P, 47 Sc、 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 111 In、 124 I, 125 I or 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 203 Pb, 212 Pb, 225 Ac.

[0143] 3. The method as described in embodiment 1, wherein the radionuclide is 64 Cu.

[0144] 4. The method according to any one of embodiments 1 to 3, wherein the first PEG portion and the second PEG portion are independently selected from: PEG 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and (PEG 28 )2 (PEG 1500x2).

[0145] 5. The method as described in any one of embodiments 1 to 4, further comprising administering one or more additional doses of the therapeutic conjugate to the subject.

[0146] 6. The method of any one of embodiments 1 to 5, wherein the dose comprises no more than about 100 µg of the peptide.

[0147] 7. The method as described in any one of embodiments 1 to 6, wherein the α v β6 integrin-related cancers include solid tumors, pancreatic cancer, breast cancer, colorectal cancer, lung cancer, ovarian cancer, cervical cancer, oral squamous cell carcinoma, skin squamous cell carcinoma, gastric cancer, or endometrial cancer.

[0148] 8. The method of embodiment 7, wherein the pancreatic cancer is locally advanced or metastatic pancreatic cancer; locally advanced, unresectable or metastatic pancreatic adenocarcinoma; or pancreatic ductal adenocarcinoma (PDAC).

[0149] 9. The method as described in any one of embodiments 1 to 8, wherein the α v β6 integrin-related cancers include primary lesions and metastatic lesions.

[0150] 10. The method as described in any one of embodiments 1 to 9, wherein the α v β6 integrin-related cancers include lesions in the adrenal glands, bone, brain, liver, lungs, or any combination of the above.

[0151] 11. The method as described in any one of embodiments 1 to 10, wherein the subject receives standard care treatment prior to administration of the therapeutic conjugate at the stated dose.

[0152] 12. The method as described in any one of embodiments 1 to 11, wherein the subject receives standard care treatment after administration of the therapeutic conjugate in the stated dose.

[0153] 13. The method as described in embodiment 11 or 12, wherein the standard care treatment includes one or more of surgery, radiation therapy, chemotherapy, radiotherapy and targeted therapy.

[0154] 14. The method of any one of embodiments 11 to 13, wherein the standard care treatment comprises FOLFIRINOX (leucovorin calcium (leucovorin), fluorouracil, irinotecan hydrochloride and oxaliplatin), gemcitabine, albumin-bound paclitaxel, irinotecan or a combination thereof.

[0155] 15. The method of any one of embodiments 1 to 14, wherein the method further comprises scanning the body or a portion thereof of the subject after application of the therapeutic conjugate.

[0156] 16. The method of embodiment 15, wherein the scan includes positron emission tomography (PET), computed tomography (CT) scan, or single-photon emission computed tomography (SPECT).

[0157] 17. The method of any one of embodiments 1 to 16, further comprising applying a diagnostic conjugate prior to applying the therapeutic conjugate, wherein the diagnostic conjugate comprises an RGD peptide and a second radionuclide.

[0158] 18. The method as described in embodiment 17, wherein the second radionuclide is 64 Cu.

[0159] 19. The method as described in embodiment 17 or 18, wherein the diagnostic conjugate is administered at a dose containing up to about 5 mCi of radioactivity.

[0160] 20. The method of any one of embodiments 17 to 19, wherein the method further comprises scanning the body or a portion thereof of the subject after application of the diagnostic conjugate.

[0161] 21. The method of embodiment 20, wherein the scan includes positron emission tomography (PET), computed tomography (CT) scan, or single-photon emission computed tomography (SPECT).

[0162] 22. The method as described in any one of embodiments 17 to 21, wherein the therapeutic conjugate is administered within 5 weeks after the administration of the diagnostic conjugate.

[0163] 23. The method of any one of embodiments 17 to 22, wherein the diagnostic conjugate administered to the subject requiring treatment comprises:

[0164] RN is a radioactive nuclide, preferably 64Cu; wherein 5G is a PEGylated peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1) and comprising a first PEG portion at the N-terminus of the peptide and a second PEG portion at the C-terminus of the peptide, and wherein ABM is an albumin-binding portion.

[0165] 24. The method of embodiment 23, wherein the first PEG portion and the second PEG portion each comprise PEG. 28 (PEG 1500).

[0166] 25. The method of any one of embodiments 1 to 24, further comprising applying an amino acid solution to the object.

[0167] 26. The method of embodiment 25, wherein the solution is applied before or simultaneously with the application of the therapeutic conjugate.

[0168] 27. The method of any one of embodiments 1 to 26, wherein the therapeutic conjugate is administered to the subject by infusion.

[0169] 28. The method of any one of embodiments 18 to 27, wherein the diagnostic conjugate is administered to the subject by injection.

[0170] 29. The method as described in any one of embodiments 1 to 28, wherein the treatment results in stable, partial or complete remission of the condition.

[0171] 30. The method of any one of embodiments 1 to 29, wherein the treatment results in a reduction of cancer metastasis in the subject.

[0172] 31. The method of any one of embodiments 1 to 30, wherein the treatment results in a reduction in tumor volume, size, or growth in the subject.

[0173] 32. The method of any one of embodiments 1 to 31, wherein the treatment results in an increased response of the cancer to a subsequently applied anticancer agent.

[0174] 33. The method of any one of embodiments 1 to 32, wherein the amount of the therapeutic conjugate present in the kidney tissue about 24 hours, about 48 hours, or about 72 hours after application of the conjugate is less than the amount of the therapeutic conjugate present in the kidney tissue 1 hour after application of the conjugate.

[0175] 34. The method as described in any one of embodiments 1 to 33, wherein the ratio of the amount of the therapeutic conjugate in the primary tumor to the amount of the therapeutic conjugate in the kidney tissue approximately 24 hours, approximately 48 hours, or approximately 72 hours after application of the conjugate is higher than the ratio of the amount of the therapeutic conjugate in the primary tumor to the amount of the therapeutic conjugate in the kidney tissue approximately 1 hour after application of the conjugate.

[0176] 35. A pharmaceutical composition comprising a peptide covalently linked to the (2,2',2”-(1,4,7-triazacyclononane-1,4,7-triyl)triacetic acid) moiety of NOTA, said peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO:1).

[0177] 36. The pharmaceutical composition of embodiment 35, wherein the PEG portion is covalently linked to the N-terminus, C-terminus, or both of the N-terminus and C-terminus of the peptide.

[0178] 37. The pharmaceutical composition of embodiment 36, wherein the PEG portion is independently selected from: PEG 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and / or (PEG 28 )2 (PEG 1500×2).

[0179] 38. The pharmaceutical composition of embodiment 36, wherein a first PEG moiety is covalently linked to the N-terminus of the peptide, a second PEG moiety is covalently linked to the C-terminus of the peptide, and the first PEG moiety and the second PEG moiety are independently selected from: PEG 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and / or (PEG 28 )2 (PEG 1500×2).

[0180] 39. The pharmaceutical composition of embodiment 37, wherein the first PEG portion and the second PEG portion are identical.

[0181] 40. The pharmaceutical composition of embodiment 38, wherein the first PEG portion and the second PEG portion each comprise PEG. 28 (PEG 1500).

[0182] 41. The pharmaceutical composition of any one of embodiments 35 to 40, wherein the peptide is covalently linked to the albumin-binding moiety (ABM).

[0183] 42. The pharmaceutical composition of embodiment 41, wherein the ABM comprises 4-(4-iodophenyl)butyric acid.

[0184] 43. The pharmaceutical composition of embodiment 41, wherein the ABM comprises a K(D-Abu-iodophenylbutyryl) moiety.

[0185] 44. The pharmaceutical composition of any one of embodiments 35 to 43, wherein the radionuclide is complexed with the NOA portion.

[0186] 45. The pharmaceutical composition of any one of embodiments 35 to 43, wherein a radionuclide is covalently linked directly or indirectly to the peptide.

[0187] 46. ​​The pharmaceutical composition of embodiment 44 or 45, wherein the radionuclide is selected from: 32 P, 47 Sc、 67 Cu、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 212 Pb and 225 Ac.

[0188] 47. The pharmaceutical composition as described in embodiment 44 or 45, wherein the radionuclide is selected from: 11 C 13 N、 15 O、 18 F, 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 111 In、 124 I, 125 I, 131 I or203 Pb.

[0189] 48. A pharmaceutical composition comprising a conjugate of formula I or formula IV and a pharmaceutically acceptable excipient: (Formula I) (Formula IV) Wherein RN is a radioactive nuclide (preferred) 67 Cu); wherein X is 5G or ABM-5G; wherein 5G is a PEGylated peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), and comprising a first PEG portion at the N-terminus of the peptide and a second PEG portion at the C-terminus of the peptide, and further optionally, wherein ABM is an albumin-binding portion; If X is ABM-5G, then the resulting therapeutic conjugate is included in Formula II: (Formula II) If X is 5G, then the resulting therapeutic conjugate is included in Formula III: (Formula III)

[0190] 49. The pharmaceutical composition of embodiment 48, wherein the first PEG portion and the second PEG portion are identical.

[0191] 50. The pharmaceutical composition of embodiment 48 or 49, wherein the first PEG portion and the second PEG portion are independently selected from: PEG 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and (PEG 28 )2 (PEG 1500x2).

[0192] 51. A method for in vivo imaging of a target tissue, the method comprising: (a) Applying to a subject a conjugate comprising a peptide covalently linked to the NOA (2,2′,2”-(1,4,7-triazacyclononane-1,4,7-triyl)triacetic acid) moiety, said peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), wherein a radionuclide is complexed with said NOA moiety; and (b) Detect the conjugate to determine where the conjugate is concentrated in the object.

[0193] 52. The method of embodiment 51, wherein the radionuclide is selected from: 11 C 13 N、 15 O、 18 F, 32 P, 47 Sc、 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 111 In、 124 I, 125 I, or 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 203 Pb, 212 Pb, 225 Ac.

[0194] 53. The method as described in embodiment 51 or 52, wherein the target tissue is cancerous tissue or organ.

[0195] 54. The method of any one of embodiments 51 to 53, wherein the developing agent is a radionuclide, and wherein radiation from the radionuclide is used to determine where the conjugate is concentrated in the object.

[0196] 55. The method of any one of embodiments 51 to 54, wherein the conjugate is detected by magnetic resonance imaging (MRI), magnetic resonance spectroscopy (MRS), single-photon emission computed tomography (SPECT), positron emission tomography (PET), or optical imaging.

[0197] 56. The method of any one of embodiments 51 to 55, wherein the conjugate is detected to diagnose or predict diseases or conditions mediated by the integrin.

[0198] 57. The method of embodiment 56, wherein the disease or symptom is associated with the expression, overexpression, or activation of the integrin.

[0199] 58. The method as described in embodiment 56 or 57, wherein the disease or symptom is α. v β6 integrin-mediated diseases or conditions.

[0200] 59. Treatment of α v A method for treating β6 integrin-associated cancer, comprising administering a dose of a therapeutic conjugate to a subject; wherein the therapeutic conjugate comprises a peptide, a chelating moiety, and a radionuclide with the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), wherein the peptide further comprises a first PEG moiety at the N-terminus of the peptide and a second PEG moiety at the C-terminus of the peptide, wherein the radionuclide is... 67 Cu.

[0201] 60. The method of embodiment 59, wherein the therapeutic conjugate further comprises an albumin-binding portion.

[0202] 61. The method as described in embodiment 59 or 60, wherein the chelating portion is NOA or DOTA.

[0203] 62. The method of any one of embodiments 59 to 61, wherein the method comprises administering more than one dose of the therapeutic conjugate to the subject.

[0204] 63. The method of embodiment 61, wherein two, three, or four doses of the therapeutic conjugate are administered to the subject.

[0205] 64. The method of any one of embodiments 59 to 63, wherein the amount of the therapeutic conjugate in the kidney of the subject is reduced by at least two-fold within approximately 24 hours after administration of the dose of the therapeutic conjugate.

[0206] It should be understood that the embodiments and implementations described herein are for illustrative purposes only, and various modifications or changes will occur to those skilled in the art based thereon, and will be included within the spirit and scope of this application and the appended claims. All publications, patents, patent applications and serial number referenced herein are incorporated herein by way of their entirety for all purposes.

Claims

1. Treatment of α v A method for treating β6 integrin-related cancers, comprising administering a dose of a therapeutic conjugate of formula I to a subject requiring treatment: (Equation I) Wherein RN is a radioactive nuclide (preferred) 67 Cu); wherein X is 5G or ABM-5G; wherein 5G is a PEGylated peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1) and comprising a first PEG portion at the N-terminus of the peptide and a second PEG portion at the C-terminus of the peptide, and further optionally, wherein ABM is an albumin-binding portion; If X is ABM-5G, then the resulting therapeutic conjugate is included in Formula II: (Formula II); If X is 5G, then the resulting therapeutic conjugate is included in Formula III: (Formula III) 2. The method of claim 1, wherein the radionuclide is selected from: 11 C 13 N、 15 O、 18 F, 32 P, 47 Sc、 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 111 In、 124 I, 125 I or 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 203 Pb, 212 Pb, 225 Ac.

3. The method of claim 1, wherein the radioactive nuclide is 67 Cu.

4. The method of any one of claims 1 to 3, wherein the first PEG portion and the second PEG portion are independently selected from: PEG 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and (PEG 28 )2 (PEG 1500x2).

5. The method of any one of claims 1 to 4, further comprising administering one or more additional doses of the therapeutic conjugate to the subject.

6. The method of any one of claims 1 to 5, wherein the dose comprises no more than about 100 µg of the peptide.

7. The method according to any one of claims 1 to 6, wherein the α v β6 integrin-related cancers include solid tumors, pancreatic cancer, breast cancer, colorectal cancer, lung cancer, ovarian cancer, cervical cancer, oral squamous cell carcinoma, skin squamous cell carcinoma, gastric cancer, or endometrial cancer.

8. The method of claim 7, wherein the pancreatic cancer is locally advanced or metastatic pancreatic cancer; locally advanced, unresectable or metastatic pancreatic adenocarcinoma; or pancreatic ductal adenocarcinoma (PDAC).

9. The method according to any one of claims 1 to 8, wherein the α v β6 integrin-related cancers include primary lesions and metastatic lesions.

10. The method of any one of claims 1 to 9, wherein the α v β6 integrin-related cancers include lesions in the adrenal glands, bone, brain, liver, lungs, or any combination of the above.

11. The method of any one of claims 1 to 10, wherein the subject receives standard care treatment prior to administration of the therapeutic conjugate in the dose stated therein.

12. The method of any one of claims 1 to 11, wherein the subject receives standard care treatment after administration of the therapeutic conjugate in the stated dose.

13. The method of claim 11 or 12, wherein the standard care treatment includes one or more of surgery, radiotherapy, chemotherapy, chemoradiotherapy, and targeted therapy.

14. The method of any one of claims 11 to 13, wherein the standard care treatment comprises FOLFIRINOX (leucovorin calcium (leucovorin), fluorouracil, irinotecan hydrochloride and oxaliplatin), gemcitabine, albumin-bound paclitaxel, irinotecan or a combination thereof.

15. The method of any one of claims 1 to 14, wherein the method further comprises scanning the body or a portion thereof of the subject after application of the therapeutic conjugate.

16. The method of claim 15, wherein the scan comprises positron emission tomography (PET), computed tomography (CT) scan, magnetic resonance imaging (MRI), or single-photon emission computed tomography (SPECT).

17. The method of any one of claims 1 to 16, further comprising applying a diagnostic conjugate prior to applying the therapeutic conjugate, wherein the diagnostic conjugate comprises an RGD peptide and a second radionuclide.

18. The method of claim 17, wherein the second radionuclide is 64 Cu.

19. The method of claim 17 or 18, wherein the diagnostic conjugate is administered at a dose containing up to about 5 mCi of radioactivity.

20. The method of any one of claims 17 to 19, wherein the method further comprises scanning the body or a portion thereof of the subject after application of the diagnostic conjugate.

21. The method of claim 20, wherein the scan comprises positron emission tomography (PET), computed tomography (CT) scan, or single-photon emission computed tomography (SPECT).

22. The method of any one of claims 17 to 21, wherein the therapeutic conjugate is administered within 5 weeks after the administration of the diagnostic conjugate.

23. The method of any one of claims 17 to 22, wherein the diagnostic conjugate administered to the subject requiring treatment comprises formula II: (Formula II) RN is a radioactive nuclide, preferably... 64 Cu; wherein 5G is a PEGylated peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1) and comprising a first PEG portion at the N-terminus of the peptide and a second PEG portion at the C-terminus of the peptide, and further optionally, wherein ABM is an albumin-binding portion.

24. The method of claim 23, wherein the first PEG portion and the second PEG portion each comprise PEG. 28 (PEG 1500).

25. The method of any one of claims 1 to 24, further comprising applying an amino acid solution to the object.

26. The method of claim 25, wherein the solution is applied before or simultaneously with the application of the therapeutic conjugate.

27. The method of any one of claims 1 to 26, wherein the therapeutic conjugate is administered to the subject by infusion.

28. The method of any one of claims 18 to 27, wherein the diagnostic conjugate is administered to the subject by injection.

29. The method of any one of claims 1 to 28, wherein the treatment results in stabilization, partial remission, or complete remission of the condition.

30. The method of any one of claims 1 to 29, wherein the treatment results in a reduction of cancer metastasis in the subject.

31. The method of any one of claims 1 to 30, wherein the treatment results in a reduction in the volume, size, or growth of the tumor in the object.

32. The method of any one of claims 1 to 31, wherein the treatment results in an increased response of the cancer to a subsequently applied anticancer agent.

33. The method of any one of claims 1 to 32, wherein the amount of the therapeutic conjugate present in the kidney tissue about 24 hours, about 48 hours, or about 72 hours after application of the conjugate is less than the amount of the therapeutic conjugate present in the kidney tissue 1 hour after application of the conjugate.

34. The method of any one of claims 1 to 33, wherein the ratio of the amount of the therapeutic conjugate in the primary tumor to the amount of the therapeutic conjugate in the kidney tissue about 24 hours, about 48 hours, or about 72 hours after application of the conjugate is higher than the ratio of the amount of the therapeutic conjugate in the primary tumor to the amount of the therapeutic conjugate in the kidney tissue about 1 hour after application of the conjugate.

35. A pharmaceutical composition comprising a peptide covalently linked to the (2,2',2”-(1,4,7-triazacyclononane-1,4,7-triyl)triacetic acid) moiety of NOTA, said peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1).

36. The pharmaceutical composition of claim 35, wherein the PEG portion is covalently linked to the N-terminus, C-terminus, or both of the N-terminus and C-terminus of the peptide.

37. The pharmaceutical composition of claim 36, wherein the PEG portion is independently selected from: PEG 11 PEG 12 (PEG800), PEG 28 (PEG 1500) and / or (PEG 28 )2 (PEG 1500×2).

38. The pharmaceutical composition of claim 36, wherein the first PEG moiety is covalently linked to the N-terminus of the peptide, the second PEG moiety is covalently linked to the C-terminus of the peptide, and the first PEG moiety and the second PEG moiety are independently selected from: PEG 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and / or (PEG 28 )2 (PEG 1500×2).

39. The pharmaceutical composition of claim 37, wherein the first PEG portion and the second PEG portion are identical.

40. The pharmaceutical composition of claim 38, wherein the first PEG portion and the second PEG portion each comprise PEG. 28 (PEG 1500).

41. The pharmaceutical composition of any one of claims 35 to 40, wherein the peptide is covalently linked to the albumin-binding moiety (ABM).

42. The pharmaceutical composition of claim 41, wherein the ABM comprises 4-(4-iodophenyl)butyric acid.

43. The pharmaceutical composition of claim 41, wherein the ABM comprises a K(D-Abu-iodophenylbutyryl) moiety.

44. The pharmaceutical composition of any one of claims 35 to 43, wherein the radionuclide is complexed with the NOA portion.

45. The pharmaceutical composition of any one of claims 35 to 43, wherein a radionuclide is covalently linked to the peptide directly or indirectly.

46. ​​The pharmaceutical composition of claim 44 or 45, wherein the radionuclide is selected from: 47 Sc、 67 Cu、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 212 Pb and 225 Ac.

47. The pharmaceutical composition of claim 44 or 45, wherein the radionuclide is selected from: 11 C 13 N、 15 O、 18 F, 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 111 In、 124 I, 125 I, 131 I or 203 Pb.

48. A pharmaceutical composition comprising a conjugate of formula I and a pharmaceutically acceptable excipient: (Equation I) Wherein RN is a radioactive nuclide (preferred) 67 Cu); wherein X is 5G or ABM-5G; wherein 5G is a PEGylated peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1) and comprising a first PEG portion at the N-terminus of the peptide and a second PEG portion at the C-terminus of the peptide, and further optionally, wherein ABM is an albumin-binding portion; If X is ABM-5G, then the resulting therapeutic conjugate is included in Formula II: (Formula II); and If X is 5G, then the resulting therapeutic conjugate is included in Formula III: (Formula III) 49. The pharmaceutical composition of claim 48, wherein the first PEG portion and the second PEG portion are identical.

50. The pharmaceutical composition of claim 48 or 49, wherein the first PEG portion and the second PEG portion are independently selected from: PEG 11 PEG 12 (PEG 800), PEG 28 (PEG 1500) and (PEG 28 )2 (PEG 1500x2).

51. A method for in vivo imaging of a target tissue, the method comprising: (a) Applying to a subject a conjugate comprising a peptide covalently linked to the NOA(2,2',2”-(1,4,7-triazacyclononane-1,4,7-triyl)triacetic acid) moiety, the peptide having the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO:1), wherein a radionuclide is complexed with the NOA moiety; as well as (b) Detect the conjugate to determine where the conjugate is concentrated in the object.

52. The method of claim 51, wherein the radionuclide is selected from: 11 C 13 N、 15 O、 18 F, 32 P, 47 Sc、 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 66 Ga、 67 Ga、 67 Cu、 68 Ga、 82 Rb、 86 Y、 89 Sr、 90 Y、 105 Rh、 111 Ag、 117m Sn、 111 In、 124 I, 125 I, or 131 I, 149 Pm, 153 Sm、 166 Ho、 177 Lu、 186 Re、 188 Re、 211 At、 212 Bi、 203 Pb, 212 Pb, 225 Ac.

53. The method of claim 51 or 52, wherein the target tissue is cancerous tissue or organ.

54. The method of any one of claims 51 to 53, wherein the developing agent is a radionuclide, and wherein radiation from the radionuclide is used to determine where the conjugate is concentrated in the object.

55. The method of any one of claims 51 to 54, wherein the conjugate is detected by magnetic resonance imaging (MRI), magnetic resonance spectroscopy (MRS), single-photon emission computed tomography (SPECT), positron emission tomography (PET), or optical imaging.

56. The method of any one of claims 51 to 55, wherein the conjugate is detected to diagnose or predict diseases or conditions mediated by the integrin.

57. The method of claim 56, wherein the disease or symptom is associated with the expression, overexpression, or activation of the integrin.

58. The method of claim 56 or 57, wherein the disease or symptom is α. v β6 integrin-mediated diseases or conditions.

59. Treatment of α v A method for treating β6 integrin-related cancer, comprising administering a dose of a therapeutic conjugate to a subject, wherein the therapeutic conjugate comprises a peptide with the amino acid sequence GNGVPNLRGDLQVLGQRVGRT (SEQ ID NO: 1), a chelating moiety, and a radionuclide, wherein the peptide further comprises a first PEG moiety at the N-terminus of the peptide and a second PEG moiety at the C-terminus of the peptide, wherein the radionuclide is... 67 Cu.

60. The method of claim 59, wherein the therapeutic conjugate further comprises an albumin-binding portion.

61. The method of claim 59 or 60, wherein the chelating portion is NOA or DOTA.

62. The method of any one of claims 59 to 61, wherein the method comprises administering more than one dose of the therapeutic conjugate to the subject.

63. The method of claim 61, wherein two, three, or four doses of the therapeutic conjugate are administered to the subject.

64. The method of any one of claims 59 to 63, wherein the amount of the therapeutic conjugate in the kidney of the subject is reduced by at least two-fold within approximately 24 hours after administration of the dose of the therapeutic conjugate.

65. The method of any one of claims 59 to 64, wherein the conjugate has the structure of formula I.

66. The method of any one of claims 59 to 64, wherein the conjugate has the structure of formula II.

67. The method of any one of claims 59 to 64, wherein the conjugate has the structure of formula III.

68. The method of any one of claims 59 to 64, wherein the conjugate has the structure of formula IV.

69. The method of any one of claims 59 to 64, further comprising imaging the tumor or tumor cells in the subject with an imaging agent, wherein the imaging agent comprises formula I, optionally wherein the radionuclide is 64 Cu.

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