Fibrin-binding compounds for imaging and therapy

Abstract

The present disclosure relates to compounds of formula IV: for use in fibrin imaging, wherein the compound comprises an imaging or therapeutic radioisotope.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 62 / 924,997, filed on October 23, 2019, which is incorporated herein by reference in its entirety.

[0003] Federally sponsored research or development

[0004] This invention was made with government support under Contract No. HHSN268201400044C awarded by the National Institutes of Health. The government has certain rights in this invention. Technical Field

[0005] The present disclosure relates to fibrin-binding compounds comprising a radioactive moiety for use in diagnostic imaging and treatment of various diseases and conditions associated with the presence of fibrin. Background Art

[0006] Fibrin is a fibrous, nonglobular protein derived from the soluble plasma protein fibrinogen and is the main component of thrombi. Fibrinogen polymerization promoted by the protease thrombin forms fibrin, which together with platelets leads to the formation of a thrombus at the wound site, thereby preventing further bleeding. Fibrin is present in all thrombi, regardless of the age or body location of the thrombus, and can be used to diagnose and treat diseases and conditions involving the presence of fibrin. Diagnostic imaging techniques such as magnetic resonance imaging (MRI), X-rays, and nuclear radiopharmaceutical imaging including positron emission tomography (PET) and single photon emission computed tomography (SPECT) are commonly used to diagnose cardiovascular conditions. One approach relies on visualization of thrombi achieved through specific molecular targets including fibrin.

[0007] In addition, fibrin is known to play an important role in the pathophysiology of malignant tumors (Costantini and Zacharski, 1992, Cancer and Metastasis Rev., 11, 283). In cancer, tumor invasion and metastasis may lead to erosion of adjacent vascular tissue and then bleeding, followed by thrombus formation in the tumor and replaced by collagen in a manner similar to the normal wound healing process (see, for example, Falanga et al., 2013, J Thromb Haemost, 11, 223-233; Obonai et al., 2016, Sci. Rep., 6, 23613). Unlike the normal wound healing process (thrombus is only formed at the beginning of the wound, and eventually disappears due to plasmin digestion or replacement by collagen), as long as the cancer cells survive in vivo, the fibrin clot in cancer will persist. The deposition of insoluble fibrin in various tumor tissues and thrombi is related to the invasiveness and progression of the tumor. Therefore, there is a need for fibrin targeting agents that can be used to diagnose and treat various cancers.

[0008] In addition, fibrin deposits are known to be associated with neurodegenerative diseases associated with systemic inflammation (neuroinflammation), such as Alzheimer's disease (AD), multiple sclerosis, and traumatic brain injury (TBI). Fibrin deposits are associated with decreased memory in neuroinflammatory diseases including AD and TBI (Sulimai and Lominadze, 2020, Mol. Neurobiol., 57, 4692-4703). Therefore, there is also a need for fibrin targeting agents that can be used to diagnose and treat neuroinflammation.

[0009] Provided herein are fibrin-specific binding compounds and methods for imaging fibrin. Also provided herein are methods for treating various diseases and conditions associated with the presence of fibrin, including cardiovascular disease, cerebrovascular disease, and cancer. Summary of the invention

[0010] The present disclosure provides compounds of formula IV:

[0011]

[0012] or a pharmaceutically acceptable salt thereof,

[0013] Among them, R 4 It is a radioactive isotope;

[0014] C 4 is a chelating moiety selected from the group consisting of:

[0015]

[0016]

[0017]

[0018]

[0019] CP 4 It is a fibrin-binding peptide;

[0020] AA is the N-terminal amino acid of the fibrin-binding peptide;

[0021] L 4 For the joint;

[0022] y is an integer selected from 0 and 1; and

[0023] z is an integer selected from 0 and 1.

[0024] In some embodiments of the compound of Formula IV, R 4 is a radioisotope selected from therapeutic radioisotopes and radioisotopes detectable using nuclear imaging techniques. In some embodiments, the radioisotope detectable using nuclear imaging techniques is a positron emitting isotope or a radioisotope suitable for single photon emission computed tomography (SPECT) imaging. In some embodiments, the positron emitting isotope is selected from fluorine-18, aluminum fluoride (Al 11 In some embodiments, the positron emitting isotope is selected from the group consisting of fluorine-18, copper-64, and gallium-68. In some embodiments, the positron emitting isotope is fluorine-18. In some embodiments, the positron emitting isotope is copper-64. In some embodiments, the positron emitting isotope is gallium-68. In some embodiments, the radioactive isotope suitable for SPECT imaging is selected from the group consisting of gallium-67, technetium-99m, indium-111, iodine-123, terbium-155, and lead-203.

[0025] In some embodiments, the radioisotope is a therapeutic radioisotope (e.g., a beta emitter or an alpha emitter). In some embodiments, the therapeutic radioisotope is selected from scandium-47, copper-67, yttrium-90, iodine-125, iodine-131, samarium-153, terbium-161, holmium-166, lutetium-177, rhenium-188, astatine-211, lead-212, bismuth-213, radium-223, actinium-225, and thorium-227. In some embodiments, the therapeutic isotope is selected from yttrium-90, lutetium-177, and actinium-225. In some embodiments, the therapeutic isotope is yttrium-90. In some embodiments, the therapeutic isotope is lutetium-177. In some embodiments, the therapeutic isotope is actinium-225.

[0026] In some embodiments of the compound of Formula IV, AA-CP 4 is a fibrin-binding peptide comprising a sequence having at least 80% sequence identity to the polypeptide of SEQ ID NO: 1:

[0027] –y*–x 1 –C–Hyp–Y(3-Cl)–X 2 –L–C–X 3 –I–X 4 – (SEQ ID NO: 1)

[0028] Among them, X 1 , X 2 , X 3 and X 4 are each independently any amino acid; and

[0029] y* is L-tyrosine or D-tyrosine.

[0030] In some embodiments of the compound of Formula IV, AA-CP 4 is a fibrin-binding peptide comprising a polypeptide having at least 80% sequence identity with a polypeptide selected from the group consisting of:

[0031]

[0032] In some embodiments of the compound of Formula IV, AA-CP 4 is a fibrin-binding peptide having at least 80% sequence identity with a polypeptide selected from the group consisting of:

[0033]

[0034] In some embodiments of the compound of Formula IV, AA-CP 4 is a fibrin-binding peptide having at least 80% sequence identity with a polypeptide selected from the group consisting of:

[0035]

[0036] In some embodiments of the compound of Formula IV, AA-CP 4 is a fibrin-binding peptide having at least 80% sequence identity to:

[0037]

[0038] In some embodiments of the compound of Formula IV, C 4 Independently selected from the group consisting of:

[0039]

[0040] In some embodiments, C 4 for In some embodiments, C 4 Independently selected from the group consisting of:

[0041]

[0042]

[0043] In some embodiments, C 4 Independently selected from the group consisting of:

[0044]

[0045] In some embodiments of the compound of formula IV, y is 0. In some embodiments, y is 1.

[0046] In some embodiments of the compound of Formula IV, L 4 It is pyridyl or (pyridyl)-C(O)-.

[0047] In some embodiments of the compound of formula IV, z is 0. In some embodiments, z is 1.

[0048] In some embodiments, y is 1 and z is 0. In some embodiments, y is 1 and z is 1. In some embodiments, y is 0 and z is 0. In some embodiments, y is 0 and z is 1.

[0049] In some embodiments, the compound of formula IV is a compound of formula IVa:

[0050]

[0051] or a pharmaceutically acceptable salt thereof, wherein R 4 is capable of being chelated by part C 4 Chelated radioisotopes.

[0052] In some embodiments, R 4 Selected from aluminum fluoride (Al 18 F), scandium-43, scandium-44, scandium-47, manganese-51, manganese-52, copper-60, copper-61, copper-62, copper-64, copper-67, gallium-67, gallium-68, yttrium-86, zirconium-89, technetium-99m, yttrium-90, indium-111, terbium-149, terbium-152, samarium-153, terbium-155, terbium-161, holmium-166, lutetium-177, rhenium-188, lead-203, lead-212, bismuth-213, radium-223, actinium-225, and thorium-227.

[0053] In some embodiments, the compound of formula IV is a compound of formula IVb:

[0054]

[0055] or a pharmaceutically acceptable salt thereof, wherein R 4 is able to be connected with the connector L 4 , the N-terminal amino acid of the fibrin binding peptide AA, or a radioactive isotope covalently bound to both.

[0056] In some embodiments, R 4 Selected from fluorine-18, iodine-123, iodine-124, iodine-125, iodine-131 and astatine-211.

[0057] In some embodiments, the compound of formula IV is a compound of formula IVc:

[0058]

[0059] or a pharmaceutically acceptable salt thereof, wherein R 4 is able to be connected with the connector L 4 , the N-terminal amino acid of the fibrin binding peptide AA, or a radioactive isotope covalently bound to both.

[0060] In some embodiments, R 4 Selected from fluorine-18, iodine-123, iodine-124, iodine-125, iodine-131 and astatine-211.

[0061] In some embodiments, the compound of formula IV is a compound of formula IVd:

[0062]

[0063] or a pharmaceutically acceptable salt thereof, wherein R 4 is able to be connected with the connector L 4 , the N-terminal amino acid of the fibrin binding peptide AA, or a radioactive isotope covalently bound to both.

[0064] In some embodiments, R 4 Selected from fluorine-18, iodine-123, iodine-124, iodine-125, iodine-131 and astatine-211.

[0065] In some embodiments, the compound of formula IV is selected from the group consisting of:

[0066]

[0067]

[0068]

[0069]

[0070] or a pharmaceutically acceptable salt thereof.

[0071] In some embodiments, the compound of formula IV is selected from the group consisting of:

[0072]

[0073] or a pharmaceutically acceptable salt thereof.

[0074] In some embodiments, the compound of formula IV is:

[0075]

[0076] or a pharmaceutically acceptable salt thereof.

[0077] In some embodiments, the compound of formula IV is selected from the group consisting of:

[0078]

[0079]

[0080] The present disclosure also provides a pharmaceutical composition comprising a compound of formula IV or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0081] In some embodiments, the pharmaceutical composition comprises a free radical scavenger. In some embodiments, the free radical scavenger is an antioxidant. In some embodiments, the free radical scavenger is selected from carnosic acid, green tea extract, apigenin, diosmin, rosmarinic acid, lipoic acid, beta-carotene, L-ascorbic acid (vitamin C), N-acetylcysteine ​​(NAC), delta-tocopherol, rutin, amifostine, resveratrol, gentisic acid and gallic acid.

[0082] Also provided herein is a method for imaging fibrin in a mammal, the method comprising: a) administering to the mammal an effective amount of a pharmaceutical composition comprising a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof, wherein R 4 is a radioactive isotope that can be detected using nuclear imaging technology; b) using nuclear imaging technology to obtain an image of mammalian fibrin; c) using magnetic resonance imaging or computed tomography to obtain an anatomical image of the mammal; and d) superimposing the images of steps b) and c) to locate the fibrin image within the anatomical image of the mammal.

[0083] In some embodiments, the presence of fibrin is associated with neuroinflammation. In some embodiments, the neuroinflammation is associated with Alzheimer's disease, multiple sclerosis, or traumatic brain injury.

[0084] In some embodiments, the method further comprises: e) administering an effective amount of a pharmaceutical composition comprising a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof, wherein R 4 It is a therapeutic radioisotope.

[0085] In some embodiments of the method, the fibrin is present in a tumor. In some embodiments, the tumor is cancerous. In some embodiments, the fibrin is present in a blood clot.

[0086] The present disclosure also provides a method for treating a disease or condition associated with fibrin in a mammal, the method comprising: administering to the mammal an effective amount of a pharmaceutical composition containing a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof, wherein R 4 It is a therapeutic radioisotope.

[0087] In some embodiments, the method further comprises administering an amino acid solution. In some embodiments, the amino acid solution comprises L-lysine, L-arginine and pharmaceutically acceptable salts thereof, and combinations thereof. In some embodiments, the amino acid solution comprises L-lysine HCl and L-arginine HCl. In some embodiments, the amino acid solution is administered before, simultaneously with, after, or a combination thereof, of a pharmaceutical composition comprising a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof. In some embodiments, the amino acid solution is administered about 30 minutes before administering a pharmaceutical composition comprising a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof. In some embodiments, the amino acid solution is administered while administering a pharmaceutical composition comprising a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof. In some embodiments, the amino acid solution is administered about 30 minutes after administering a pharmaceutical composition comprising a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof.

[0088] In some embodiments, the method further comprises administering an antiemetic. In some embodiments, the antiemetic is administered before, simultaneously, after, or a combination thereof, of a pharmaceutical composition comprising a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof. In some embodiments, the antiemetic is administered before, simultaneously, after, or a combination thereof, of an amino acid solution. In some embodiments, the antiemetic is selected from a 5-HT3 receptor antagonist, a corticosteroid, a neurokinin-1 (NK-1) receptor inhibitor, prochlorperazine, metoclopramide, and a cannabinoid.

[0089] In some embodiments of this method, the disease or condition associated with the presence of fibrin is cancer.

[0090] The present invention also provides a method for treating cancer in a mammal, the method comprising: administering to the mammal an effective amount of a pharmaceutical composition containing a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof, wherein R 4 is a therapeutic radioisotope. In some embodiments, R 4 is selected from scandium-47, copper-67, yttrium-90, iodine-131, samarium-153, terbium-161, holmium-166, lutetium-177, rhenium-188, astatine-211, lead-212, bismuth-213, radium-223, actinium-225, and thorium-227. 4 is selected from yttrium-90, lutetium-177 and actinium-225. In some embodiments, the therapeutic radioisotope R 4 In some embodiments, the therapeutic radioisotope R 4is lutetium-177. In some embodiments, R 4 It is actinium-225.

[0091] Also provided herein is a method for detecting and treating a disease or condition associated with fibrin in a mammal, the method comprising: a) administering to the mammal an effective amount of a pharmaceutical composition comprising a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof, wherein R 4 is a radioactive isotope that can be detected using nuclear imaging techniques; b) using nuclear imaging techniques to obtain an image of mammalian fibrin; c) using magnetic resonance imaging or computed tomography to obtain an anatomical image of the mammal; d) superimposing the images of steps b) and c) to locate the fibrin image within the anatomical image of the mammal; and e) administering an effective amount of a pharmaceutical composition comprising a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof, wherein R 4 It is a therapeutic radioisotope.

[0092] In some embodiments of the method, R is a radioactive isotope that can be detected using nuclear imaging techniques. 4 Selected from fluorine-18, aluminum fluoride (Al 18 F), scandium-43, scandium-44, copper-64, gallium-68, yttrium-86, zirconium-89, indium-111, iodine-123, iodine-124, terbium-149, terbium-152, terbium-155, and lead-203. In some embodiments, R that can be detected using nuclear imaging techniques 4 is selected from the group consisting of fluorine-18, copper-64, and gallium-68. In some embodiments, R detectable using nuclear imaging techniques 4 is fluorine-18. In some embodiments, R can be detected using nuclear imaging techniques. 4 is copper-64. In some embodiments, R can be detected using nuclear imaging techniques. 4 It is gallium-68.

[0093] In some embodiments of this method, R 4 is selected from scandium-47, copper-67, yttrium-90, iodine-131, samarium-153, terbium-161, holmium-166, lutetium-177, rhenium-188, astatine-211, lead-212, bismuth-213, radium-223, actinium-225, and thorium-227. 4 is selected from yttrium-90, lutetium-177 and actinium-225. In some embodiments, the therapeutic radioisotope R 4 In some embodiments, the therapeutic radioisotope R 4is lutetium-177. In some embodiments, R 4 It is actinium-225.

[0094] In some embodiments, the method further comprises administering an amino acid solution. In some embodiments, the amino acid solution is administered before, simultaneously with, after, or a combination thereof, of a pharmaceutical composition comprising a compound of Formula IV or an effective amount of a compound of Formula IV or a pharmaceutically acceptable salt thereof.

[0095] In some embodiments, the method further comprises administering an antiemetic. In some embodiments, the antiemetic is administered before, simultaneously with, after, or a combination thereof, of the amino acid solution.

[0096] In some embodiments of this method, the disease or condition associated with the presence of fibrin is cancer.

[0097] The present disclosure also provides a method for detecting and treating a disease or condition associated with fibrin in a mammal, the method comprising: a) administering to the mammal an effective amount of a pharmaceutical composition containing a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof, wherein R 4 is a radioactive isotope detectable using nuclear imaging techniques, selected from fluorine-18, copper-64 and gallium-68; b) using nuclear imaging techniques to obtain an image of mammalian fibrin; c) using magnetic resonance imaging or computed tomography to obtain an anatomical image of the mammal; d) superimposing the images of steps b) and c) to locate the fibrin image within the anatomical image of the mammal; and e) administering an effective amount of a pharmaceutical composition comprising a compound of formula IV or an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof, wherein R 4 is a therapeutic radioisotope selected from the group consisting of yttrium-90, lutetium-177, and actinium-225. In some embodiments, the fibrin is present in the tumor. In some embodiments, the tumor is cancerous.

[0098] The present disclosure also provides compounds of formula V:

[0099]

[0100] or a pharmaceutically acceptable salt thereof, wherein:

[0101] C 4 is the chelating part;

[0102] CP 4 It is a fibrin-binding peptide;

[0103] AA is the N-terminal amino acid of the fibrin-binding peptide;

[0104] L 4 For the joint;

[0105] y is an integer selected from 0 or 1; and

[0106] z is an integer selected from 0 or 1.

[0107] In some embodiments, the compound of formula V is a compound selected from the group consisting of:

[0108] Compound sequence 16 <![CDATA[NODAGA–Y–e–C–Hyp–Y(3–Cl)–G–L–C–Y–I–Q–NH2]]> 17 <![CDATA[NODAGA–y–e–C–Hyp–Y(3–Cl)–G–L–C–Y–I–Q–NH2]]> 18 <![CDATA[NODAGA–Y–e–C–Hyp–Y(3–Cl)–G–L–C–H–I–Q–NH2]]> 19 <![CDATA[NODAGA–y–e–C–Hyp–Y(3–Cl)–G–L–C–H–I–Q–NH2]]> 20 <![CDATA[NODAGA–Y–e–C–Hyp–Y(3–Cl)–G–L–C–H–I–q–NH2]]> 21 <![CDATA[NODAGA–y–e–C–Hyp–Y(3–Cl)–G–L–C–H–I–I–NH2]]> 22 <![CDATA[NODAGA-y-e-c-Hyp-Y(3-Cl)-G-L-C-H-I-q-NH2]]> 23 <![CDATA[DOTAGA–Y–e–C–Hyp–Y(3–Cl)–G–L–C–H–I–q–NH2]]> 24 NOTA–Y–e–C–Hyp–Y(3–Cl)–G–L–C–H–I–q–NH2.

[0109] Provided herein are compounds of Formula I:

[0110] [M 1 ] m –[C 1 ] n –[CP 1 ]–[L 1 ] o –[C 1 ] p –[M 1 ] q (I)

[0111] or a pharmaceutically acceptable salt thereof,

[0112] Among them, each M 1 independently copper-64 or gallium-68;

[0113] Each C 1 is a chelating moiety independently selected from the group consisting of:

[0114]

[0115] CP 1 It is a fibrin-binding peptide;

[0116] Each L 1 independently a linker moiety;

[0117] m is an integer selected from 0-5;

[0118] n is an integer selected from 0-5;

[0119] o is an integer selected from 0-5;

[0120] p is an integer selected from 0-5; and

[0121] q is an integer selected from 0-5.

[0122] In some embodiments, M 1 is copper-64. In some embodiments, M 1 It is gallium-68.

[0123] In some embodiments, each C1 Independently selected from the group consisting of:

[0124]

[0125] In some embodiments, each C 1 for

[0126] In some embodiments, each C 1 Independently selected from the group consisting of:

[0127]

[0128] In some embodiments, CP 1 is a fibrin-binding peptide comprising a sequence having at least 80% sequence identity to the polypeptide of SEQ ID NO: 1:

[0129] –y*–x 1 –C–Hyp–Y(3-Cl)–X 2 –L–C–X 3 –I–X 4 – (SEQ ID NO: 1)

[0130] Among them, X 1 , X 2 , X 3 and X 4 are each independently any amino acid; and

[0131] y* is L-tyrosine or D-tyrosine.

[0132] In some embodiments, CP 1 is a fibrin-binding peptide comprising a polypeptide having at least 80% sequence identity with a polypeptide selected from the group consisting of:

[0133]

[0134] In some embodiments, CP 1 is a fibrin-binding peptide having at least 80% sequence identity with a polypeptide selected from the group consisting of:

[0135]

[0136]

[0137] In some embodiments, each L 1 Independently selected from the group consisting of:

[0138]

[0139] In some embodiments, m is 1. In some embodiments, p is 1. In some embodiments, n is 1. In some embodiments, o is 1.

[0140] In some embodiments, the compound of formula I is selected from the group consisting of:

[0141]

[0142]

[0143]

[0144]

[0145]

[0146] or a pharmaceutically acceptable salt thereof.

[0147] Also provided herein are compounds of formula II:

[0148] [M 2 ] r –[C 2 ] s –[CP 2 ]–[R 2 ] t (II)

[0149] or a pharmaceutically acceptable salt thereof,

[0150] Among them, each M 2 independently actinium-225, astatine-211, bismuth-213, copper-64, copper-67, aluminum fluoride (AL 18 F), gallium-68, holmium-166, indium-111, iodine-123, iodine-124, iodine-131, lead-203, lead-212, lutetium-177, radium-223, samarium-153, scandium-43, scandium-44, scandium-47, terbium-149, terbium-152, terbium-155, terbium-161, thorium-227; yttrium-86, yttrium-90 or zirconium-89;

[0151] Each C 2 independently a chelating moiety;

[0152] CP 2 It is a fibrin-binding peptide;

[0153] Each R 2 independently organic non-chelating moieties;

[0154] r is an integer selected from 0-5;

[0155] s is an integer selected from 0-5; and

[0156] t is an integer selected from 0-5.

[0157] In some embodiments, M 2 is copper-64. In some embodiments, M 2 It is gallium-68.

[0158] In some embodiments, CP 2 is a fibrin-binding peptide comprising a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 16:

[0159] –y*–x 5 –C–Hyp–Y(3-Cl)–X 6 –L–C–X 7 –I–X 8 – (SEQ ID NO: 16)

[0160] Among them, X 5 , X 6 , X 7 and X 8 are each independently any amino acid; and

[0161] y* is L-tyrosine or D-tyrosine.

[0162] In some embodiments, CP 2 is a fibrin-binding peptide comprising a polypeptide having at least 80% sequence identity with a polypeptide selected from the group consisting of:

[0163]

[0164] In some embodiments, CP 2 is a fibrin-binding peptide having at least 80% sequence identity with a polypeptide selected from the group consisting of:

[0165]

[0166] In some embodiments, r is 1. In some embodiments, s is 1. In some embodiments, t is 1.

[0167] Also provided herein are compounds of formula III:

[0168] [R 3 ] u –[CP 3 ]–[C 3 ] v –[M 3 ] w (III)

[0169] or a pharmaceutically acceptable salt thereof,

[0170] Among them, each M 3 independently copper-64 or gallium-68;

[0171] Each C 3 is the chelating part;

[0172] CP 3 It is a fibrin-binding peptide;

[0173] Each R3 is independently an organic non-chelating moiety;

[0174] u is an integer selected from 0-5;

[0175] v is an integer selected from 0-5; and

[0176] w is an integer selected from 0-5.

[0177] In some embodiments, M 3 is copper-64. In some embodiments, M 3 It is gallium-68.

[0178] In some embodiments, CP 3 is a fibrin-binding peptide comprising a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 17:

[0179] –y*–x 9 –C–Hyp–Y(3-Cl)–X 10 –L–C–X 11 –I–X 12 – (SEQ ID NO: 17)

[0180] Among them, X 9 , X 10 , X 11 and X 12 are each independently any amino acid; and

[0181] y* is L-tyrosine or D-tyrosine.

[0182] In some embodiments, CP 3 is a fibrin-binding peptide comprising a sequence having at least 80% sequence identity with a polypeptide selected from the group consisting of:

[0183]

[0184] In some embodiments, CP 3 is a fibrin-binding peptide having at least 80% sequence identity with a polypeptide selected from the group consisting of:

[0185]

[0186] In some embodiments, u is 1. In some embodiments, v is 1. In some embodiments, w is 1.

[0187] Also provided herein are pharmaceutical compositions comprising a compound of Formula I, Formula II, or Formula III, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0188] Also provided herein is a method for imaging fibrin in a mammal, the method comprising administering to the mammal an effective amount of a compound of Formula I, Formula II or Formula III or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of Formula I, Formula II or Formula III or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient; obtaining an image of fibrin in the mammal using nuclear imaging technology; obtaining an anatomical image of the mammal using magnetic resonance imaging or computed tomography; and superimposing the images to locate the image of fibrin within the anatomical image of the mammal.

[0189] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference, as if each individual publication, patent, or patent application was specifically and individually incorporated herein by reference. In the event that a publication or patent or patent application incorporated by reference conflicts with the disclosure contained in the specification, the specification shall supersede and / or take precedence over any such conflicting material.

[0190] Other features and advantages of the present invention will become readily apparent from the following detailed description, the accompanying drawings and the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0191] Figure 1 Fluorescence polarization DD(E) binding / displacement assays for compounds 4, 6, 7, 10, 12, 13, 14, and 15 are described.

[0192] Figure 2 Fluorescence polarization DD(E) binding / displacement assays for compounds 2, 3, 5, 8, 9, and 11 are described.

[0193] Figure 3 The stability of the compounds in rat plasma at 37°C for up to 4 hours is described.

[0194] Figure 4 Semi-preparative HPLC purification is described 18 RadioHPLC trace of F-Py-TFP.

[0195] Figure 5 Semi-preparative HPLC purification of the compound is described 18 RadioHPLC trace of F-7.

[0196] Figure 6 Depicted are plate assay data plotted on a linear scale to highlight the saturation of approximately 2 binding sites per fibrin monomer in TBS buffer (top) and human plasma (bottom).

[0197] Figure 7 Describes the compound 18 F-7 was lysed in TBS buffer (top, K d =1.6±0.2 μM) and human plasma (bottom, K d =1.8±0.2 μM) in human fibrin.

[0198] Figure 8 Describes the compound 68 RadioHPLC trace of Ga-20.

[0199] Fig. 9 Blood clearance in a rat model of carotid artery crush injury is described. Blood samples were drawn before and 2, 5, 10, 15, 30, 60, 90, 120 minutes after probe injection. The activity in each sample was calculated as the percentage of injected dose (%ID) per gram of tissue.

[0200] Fig.10 Describes the binding of fibrin to rats before (T = 0 min) and after (T = 10, 60) injection. 68 Percentage of Ga-labeled compounds.

[0201] Fig.11 The fraction of circulating radioactivity is described. Blood collected 10 and 60 minutes after probe injection was centrifuged to separate plasma. Afterwards, the plasma was incubated in the wells immobilized with fibrin for 2 hours at room temperature. After incubation, counts in the supernatant of the fibrin-containing wells and the empty wells were measured on a gamma counter and divided by the plasma weight to determine the concentrations of unbound probe [unbound] and total probe [total], respectively. 68 The amount of Ga [bound] was calculated as [bound] = [total] - [unbound]. As a positive control, an aliquot of the dose was spiked into plasma and used to estimate the total possible fibrin binding in the assay (% bound at t = 0). The amount of functional probe in the blood at time t was calculated by taking the ratio of the % fibrin binding at time t to the % bound at t = 0 and multiplying this ratio by the total amount measured in the blood. 68 Ga%ID / g was used for determination.

[0202] Fig.12 Describes the injection of compounds in rats 68 Ga-16, 68 Ga-17, 68Ga-18, 68 Ga-19, 68 Ga-20 and 68 RadioHPLC traces of blood analysis after Ga-21. The traces represent blood analysis 15 and 90 minutes after injection.

[0203] Fig.13 Describes the injection 68 Biodistribution of Ga-labeled compounds in rats. The activities in various organs are shown as the percentage of injected dose (%ID) per gram of tissue.

[0204] Fig.14 Representative images of autoradiographs are depicted showing compound in clot (ipsi) compared to the contralateral side of the subject. 68 Ga-19 (left) and 68 Activity of Ga-20 (right).

[0205] Fig.15 (Top) Orthogonal CT images of a rat with right carotid artery thrombosis are depicted. The yellow arrow shows the location of the right carotid artery, which is slightly highlighted due to CT contrast agent perfusion. The orange arrow in the axial image (top left) shows the contralateral carotid artery. (Bottom) 68 PET-CT fusion images after Ga-20, with PET images presented in color. The green crosshairs indicate the location of the three orthogonal image slices shown, in this case centered on the right carotid artery thrombus. In this animal model, the right common carotid artery was isolated by making an incision in the throat, followed by a crush injury to the vessel to induce thrombosis. This model also results in the formation of microthrombi around the surgical injury site, which is indicated by the red arrows in the axial (lower left) and sagittal (lower right) images.

[0206] Fig.16 Describes the compound 68 Ga-20 PET uptake over time in control rabbits and rabbits with plaque rupture. The solid black line indicates the rupture:control ratio at each time point.

[0207] Fig.17 Compounds from rabbits with plaque rupture (left) and control rabbits (right) are depicted 68 Representative images of Ga-20 PET (upper), high-resolution T2 MR (middle), and TOF (lower). Arrows indicate the abdominal aorta (green) and inferior vena cava (blue). Insets show zoomed PET-MR images of the aorta and vena cava.

[0208] Fig.18 Describes the fibrin binding probe in samples from patients undergoing carotid endarterectomy 68 The uptake of Ga-20 was significantly higher than that of the non-binding probe68 Ga-22. From a high ( Fig.18 A-18F) and Low ( Fig.18 Representative autoradiogram of patient specimens with 68Ga-20 uptake (G-18L) Fig.18 A-18B and 18G-18H) and Carstairs-stained sections ( Fig.18 C-18F and 18I-18L). High uptake samples ( Fig.18 C-18F) shows the strong presence of fibrin (yellow arrows) with or without accompanying erythrocytes (green arrows) and even the formation of a fibrin mesh ( Fig.18 C-18D), while in the low uptake samples ( Fig.18 I-18L), the presence of fibrin is minimal. Scale bar: Fig.18 C, 18E, 18I, 18K: 100μm; Fig.18 200 μm in D, 18F, 18J, and 18L. Autoradiographs of discarded endarterectomy specimens from all 12 patients ( Fig.18 M) and functional probe assays ( Fig.18 N) revealed that compared with the non-binding probe 68 Ga-22, a fibrin binding probe 68 Ga-20 uptake was variable and significantly increased (P<0.05). Dashed line: 68 Ga-22 mean, mean ± SD, and mean ± 2SD cutoff. DETAILED DESCRIPTION OF THE INVENTION

[0210] Thromboembolism plays a causative role in many potentially fatal cardiovascular conditions including stroke, coronary artery conditions, deep vein thrombosis, and pulmonary embolism. Of the nearly 795,000 strokes that occur each year in the United States, more than 80% are ischemic or thromboembolic. Treatment options are largely influenced by the anatomical location of the clot. Currently, multiple tests are required to evaluate various body areas, for example, CT scans are used to locate pulmonary clots, ultrasound tests can be used for the carotid arteries, and MRI analysis provides images of the heart chambers. Rapid localization of the culprit clot is essential to determine the appropriate treatment.

[0211] Many methods have been developed to visualize thrombi throughout the body. Fibrin is a particularly attractive target because it is present in all thrombi, including arteries, veins, and the heart; it is not found in plasma, which makes visualization techniques highly specific; it can be used at all active stages of clot development; and it is a high-concentration target, with concentrations of approximately 20-100 μM. For example, fibrin-binding peptides can be functionalized with chelating moieties capable of chelating radioisotopes of various metals, including but not limited to actinium-225, bismuth-213, copper-64, copper-67, aluminum fluoride (ALF), and tantalum fluoride (T). 18 F), gallium-68, holmium-166, indium-111, lead-203, lead-212, lutetium-177, radium-223, samarium-153, scandium-43, scandium-44, scandium-47, terbium-149, terbium-152, terbium-155, terbium-161, thorium-227, yttrium-86, yttrium-90, and zirconium-89. Fibrin-binding peptides can also be radioactively radiated with radioactive isotopes (including but not limited to 18 F. 123 I. 124 I. 131 I and 211 In some embodiments, the present invention provides a fibrin-specific compound comprising one or more radioisotopes. Also provided herein is a method for imaging fibrin. Also provided herein is a method for treating a disease or condition using a fibrin-specific compound of the present invention as an imaging or diagnostic agent, a therapeutic agent, or both.

[0212] definition

[0213] Commonly used chemical abbreviations not expressly defined in this disclosure may be found in The American Chemical Society Style Guide, 2nd Edition, American Chemical Society, Washington, D.C. (1997); "2001 Guidelines for Authors" J.Org.Chem. 66(1), 24A(2001); and "A Short Guide to Abbreviations and Their Use in Peptide Science," J. Peptide Sci. 5,465-471(1999).

[0214] As used herein, the term "peptide" refers to an amino acid chain of about 2 to about 25 amino acid residues in length. All peptide sequences herein are written from N-terminus to C-terminus. For any peptides described herein containing two or more cysteine ​​residues, it should be understood that the cysteine ​​residues can form one or more disulfide bonds under non-reducing conditions. The formation of disulfide bonds can result in the formation of cyclic peptides.

[0215] As used herein, the term "natural" or "naturally occurring" amino acid refers to one of the twenty most common amino acids found in nature. Natural amino acids that have been modified to provide a label for detection purposes (e.g., a radiolabel, an optical label, or a dye) are considered natural amino acids. Natural L amino acids are represented by their standard one or three letter abbreviations. D amino acids use the lowercase convention for the standard single letter abbreviation and the "D-" prefix convention for the standard three letter abbreviation.

[0216] As used herein, the terms "chelator," "chelating group," and "chelating moiety" refer to a polydentate (multi-bonded) ligand that can form two or more separate coordination bonds between the ligand and a single central atom (usually a metal ion). In some embodiments, the metal ion is a radioactive isotope of a metal. Examples of such metal ions include, but are not limited to, actinium-225, bismuth-213, copper-64, copper-67, gallium-68, holmium-166, indium-111, lead-203, lead-212, lutetium-177, radium-223, samarium-153, scandium-43, scandium-44, scandium-47, terbium-149, terbium-152, terbium-155, terbium-161, thorium-227, yttrium-86, yttrium-90, and zirconium-89.

[0217] As used herein, the terms "radioisotope," "radioisotope," "radionuclide," and "radionuclide" are used interchangeably and refer to an unstable atom with excess nuclear energy; this excess energy can be emitted in one of three ways: as gamma radiation emitted from the nucleus; as the transfer and release of one of its electrons as a conversion electron; or as the emission of a new particle from the nucleus (an alpha or beta particle). This process is known as the radioactive decay of the radioisotope. Radioisotopes can be used for diagnostic imaging and treatment of a variety of diseases and conditions, including those described in the present disclosure.

[0218] As used herein, the terms "target binding" and "binding" refer to non-covalent interactions of a peptide or composition within a target. These non-covalent interactions are independent of each other and may be, among others, hydrophobic, hydrophilic, dipole-dipole, π-stacking, hydrogen bonding, electrostatic associations and / or Lewis acid-base interactions. The binding affinity for a target is measured as the equilibrium dissociation constant "K" for the target under a defined set of conditions. d "express.

[0219] As used herein, the term "purified" means that a peptide or compound has been separated from naturally occurring organic molecules with which it is normally associated, or, for chemically synthesized molecules, from other organic molecules present in the chemical synthesis. Typically, a polypeptide or compound is considered "purified" when it is at least 70% (e.g., at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%) free of any other proteins or organic molecules by dry weight. The terms "purified" and "isolated" are used interchangeably herein.

[0220] In the context of two or more nucleic acids or polypeptides, the term "percent identity" or "identity" refers to two or more sequences that are identical or have a specified percentage of identical nucleotides or amino acid residues. The percent identity can be measured using sequence comparison software or algorithms or by visual inspection.

[0221] Typically, the sequence identity percentage is calculated by determining the number of matching positions in the compared nucleic acid or polypeptide sequence, dividing the number of matching positions by the total number of compared nucleotides or amino acids, respectively, and then multiplying by 100. Matching positions refer to the positions where the same nucleotide or amino acid appears in the same position in the compared sequence. The total number of compared nucleotides or amino acids refers to the minimum number of nucleotides or amino acids necessary for comparing the second sequence, and does not include comparisons with non-fibrin binding sequences (e.g., forced comparisons). The total number of compared nucleotides or amino acids can correspond to the entire sequence or can correspond to a fragment of the full-length sequence.

[0222] Sequences can be aligned using the algorithm described by Altschul et al. (Nucleic Acids Res, 25:3389-3402, 1997), which is incorporated into the BLAST (Basic Local Alignment Search Tool) program, which can be obtained on the World Wide Web at ncbi.nlm.nih.gov. BLAST searches or alignments can be performed using the algorithm of Altschul et al. to determine the percent sequence identity between a nucleic acid or polypeptide and any other sequence or portion thereof. BLASTN is a program used to align and compare the identity between nucleic acid sequences, while BLASTP is a program used to align and compare the identity between amino acid sequences. When the BLAST program is used to calculate the percent identity between a fibrin binding sequence and another sequence, the default parameters of each program are used.

[0223] Where values ​​are described as ranges, it should be understood that such disclosure includes disclosure of all possible sub-ranges within such ranges as well as specific values ​​falling within such ranges, regardless of whether specific values ​​or specific sub-ranges have been explicitly stated.

[0224] Compound

[0225] Provided herein are compounds of Formula I:

[0226] [M 1 ] m –[C 1 ] n –[CP 1 ]–[L 1 ] o –[C 1 ] p –[M 1 ] q (I)

[0227] or a pharmaceutically acceptable salt thereof,

[0228] Among them, each M 1 independently copper-64 or gallium-68;

[0229] Each C 1 is a chelating moiety independently selected from the group consisting of:

[0230]

[0231]

[0232] CP 1 It is a fibrin-binding peptide;

[0233] Each L 1 independently a linker moiety;

[0234] m is an integer selected from 0-5;

[0235] n is an integer selected from 0-5;

[0236] o is an integer selected from 0-5;

[0237] p is an integer selected from 0-5; and

[0238] q is an integer selected from 0-5.

[0239] In some embodiments of Formula I, each M 1 is copper-64. In some embodiments of Formula I, each M 1 It is gallium-68.

[0240] In some embodiments of Formula I, each C 1 Independently selected from the group consisting of:

[0241]

[0242] In some embodiments of Formula I, C 1 For NODAGA:

[0243] In some embodiments of Formula I, each C 1 Independently selected from the group consisting of:

[0244]

[0245] In some embodiments of Formula I, CP 1 is a fibrin-binding peptide comprising a sequence having at least 80% sequence identity to the polypeptide of SEQ ID NO: 1:

[0246] –y*–x 1 –C–Hyp–Y(3-Cl)–X 2 –L–C–X 3 –I–X 4 – (SEQ ID NO: 1)

[0247] Among them, X 1 , X 2 , X 3 and X 4 Each is independently any amino acid; and y* is L-tyrosine or D-tyrosine. 1 is a fibrin-binding peptide comprising a sequence having at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to the polypeptide of SEQ ID NO: 1. In some embodiments, CP 1 The polypeptide is SEQ ID NO: 1 (ie it has 100% sequence identity).

[0248] In some embodiments, y* is L-tyrosine. In some embodiments, y* is D-tyrosine.

[0249] In some embodiments, X 1 , X 2 , X 3 and X 4 Each is independently selected from naturally occurring amino acids. For example, X 1 , X 2 , X 3 and X 4 Each is independently selected from Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp and Tyr. In some embodiments, X 1 , X 2 , X 3 and X 4Each is independently selected from the D-configuration of naturally occurring amino acids. 1 , X 2 , X 3 , X 4 Each is independently selected from D-Ala, D-Cys, D-Asp, D-Glu, D-Phe, D-His, D-Ile, D-Lys, D-Leu, D-Met, D-Asn, D-Pro, D-Gln, D-Arg, D-Ser, D-Thr, D-Val, D-Trp and D-Tyr. In some embodiments, X 1 In some embodiments, X 1 In some embodiments, X 2 In some embodiments, X 2 In some embodiments, X 2 In some embodiments, X 3 In some embodiments, X 3 In some embodiments, X 4 In some embodiments, X 4 In some embodiments, X 4 In some embodiments, X 4 It is D-Leu.

[0250] In some embodiments, X 1 , X 2 , X 3 and X 4 Each is independently selected from non-naturally occurring amino acids. 1 , X 2 , X 3 and X 4 Each is independently selected from Hyp, D-Hyp, Tyr-3-Cl and D-Tyr-3-Cl.

[0251] In some embodiments of Formula I, CP 1 A fibrin-binding peptide comprising a polypeptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0252]

[0253]

[0254] In some embodiments of Formula I, CP 1A fibrin-binding peptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0255]

[0256]

[0257] In some embodiments of Formula I, each L 1 Independently selected from the group consisting of:

[0258]

[0259] In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, o is 1. In some embodiments, o is 2. In some embodiments, q is 1. In some embodiments, q is 2. In some embodiments, m, p, n, o, and q are each 1. In some embodiments, m, p, n, o, and q are each 2.

[0260] In some embodiments of Formula I, the compound is selected from the group consisting of:

[0261]

[0262]

[0263]

[0264] or a pharmaceutically acceptable salt thereof.

[0265] In some embodiments of Formula I, the compound is selected from the group consisting of:

[0266]

[0267]

[0268]

[0269] or a pharmaceutically acceptable salt thereof.

[0270] Also provided herein are compounds of Formula Ia:

[0271] [M 1a ]–[C 1a ]–[CP 1a ]–[M 1a ] (Ia)

[0272] or a pharmaceutically acceptable salt thereof,

[0273] Among them, each M 1a independently copper-64 or gallium-68;

[0274] C 1a is a chelating moiety independently selected from the group consisting of:

[0275]

[0276]

[0277] And CP 1a It is a fibrin-binding peptide.

[0278] In some embodiments of Formula Ia, each M 1a is copper-64. In some embodiments of Formula Ia, each M 1a It is gallium-68.

[0279] In some embodiments of Formula Ia, C 1a For NODAGA:

[0280] In some embodiments of Formula Ia, CP 1a is a fibrin-binding peptide comprising a sequence having at least 80% sequence identity to the polypeptide of SEQ ID NO: 1:

[0281] –y*–x 1 –C–Hyp–Y(3-Cl)–X 2 –L–C–X 3 –I–X 4 – (SEQ ID NO: 1)

[0282] Among them, X 1 , X 2 , X 3 and X 4 Each is independently any amino acid; y* is L-tyrosine or D-tyrosine. 1a is a fibrin-binding peptide comprising a sequence having at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to the polypeptide of SEQ ID NO: 1. In some embodiments, CP 1a The polypeptide is SEQ ID NO: 1 (ie it has 100% sequence identity).

[0283] In some embodiments, y* is L-tyrosine. In some embodiments, y* is D-tyrosine.

[0284] In some embodiments, X 1 , X 2 , X 3 and X 4 Each is independently selected from naturally occurring amino acids. For example, X 1 , X 2 , X 3 and X 4 Each is independently selected from Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp and Tyr. In some embodiments, X 1 , X 2 , X 3 and X 4 Each is independently selected from the D-configuration of naturally occurring amino acids. 1 , X 2 , X 3 , X 4 Each is independently selected from D-Ala, D-Cys, D-Asp, D-Glu, D-Phe, D-His, D-Ile, D-Lys, D-Leu, D-Met, D-Asn, D-Pro, D-Gln, D-Arg, D-Ser, D-Thr, D-Val, D-Trp and D-Tyr. In some embodiments, X 1 In some embodiments, X 1 In some embodiments, X 2 In some embodiments, X 2 In some embodiments, X 2 In some embodiments, X 3 In some embodiments, X 3 In some embodiments, X 4 In some embodiments, X 4 In some embodiments, X 4 In some embodiments, X 4 It is D-Leu.

[0285] In some embodiments, X 1 , X 2 , X 3 and X 4 Each is independently selected from non-naturally occurring amino acids. 1 , X 2 , X 3and X 4 Each is independently selected from Hyp, D-Hyp, Tyr-3-Cl and D-Tyr-3-Cl.

[0286] In some embodiments of Formula Ia, CP 1a A fibrin-binding peptide comprising a polypeptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0287]

[0288]

[0289] In some embodiments of Formula Ia, CP 1a A fibrin-binding peptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0290]

[0291] Also provided herein are compounds of Formula Ib:

[0292] [M 1b ]–[C 1b ]–[CP 1b ]–[M 1b ] (Ib)

[0293] or a pharmaceutically acceptable salt thereof,

[0294] Among them, each M 1b independently copper-64 or gallium-68;

[0295] C 1b For NODAGA: as well as

[0296] CP 1b It is a fibrin-binding peptide.

[0297] In some embodiments of Formula Ib, each M 1b is copper-64. In some embodiments of Formula Ib, each M 1b It is gallium-68.

[0298] In some embodiments of Formula Ib, CP 1b is a fibrin-binding peptide comprising a sequence having at least 80% sequence identity to the polypeptide of SEQ ID NO: 1:

[0299] –y*–x1 –C–Hyp–Y(3-Cl)–X 2 –L–C–X 3 –I–X 4 – (SEQ ID NO: 1)

[0300] Among them, X 1 , X 2 , X 3 and X 4 Each is independently any amino acid; y* is L-tyrosine or D-tyrosine. 1b is a fibrin-binding peptide comprising a sequence having at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to the polypeptide of SEQ ID NO: 1. In some embodiments, CP 1b The polypeptide is SEQ ID NO: 1 (ie it has 100% sequence identity).

[0301] In some embodiments, y* is L-tyrosine. In some embodiments, y* is D-tyrosine.

[0302] In some embodiments, X 1 , X 2 , X 3 and X 4 Each is independently selected from naturally occurring amino acids. For example, X 1 , X 2 , X 3 and X 4 Each is independently selected from Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp and Tyr. In some embodiments, X 1 , X 2 , X 3 and X 4 Each is independently selected from the D-configuration of naturally occurring amino acids. 1 , X 2 , X 3 , X 4 Each is independently selected from D-Ala, D-Cys, D-Asp, D-Glu, D-Phe, D-His, D-Ile, D-Lys, D-Leu, D-Met, D-Asn, D-Pro, D-Gln, D-Arg, D-Ser, D-Thr, D-Val, D-Trp and D-Tyr. In some embodiments, X 1 In some embodiments, X 1In some embodiments, X 2 In some embodiments, X 2 In some embodiments, X 2 In some embodiments, X 3 In some embodiments, X 3 In some embodiments, X 4 In some embodiments, X 4 In some embodiments, X 4 In some embodiments, X 4 It is D-Leu.

[0303] In some embodiments, X 1 , X 2 , X 3 and X 4 Each is independently selected from non-naturally occurring amino acids. 1 , X 2 , X 3 and X 4 Each is independently selected from Hyp, D-Hyp, Tyr-3-Cl and D-Tyr-3-Cl.

[0304] In some embodiments of Formula Ib, CP 1b A fibrin-binding peptide comprising a polypeptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0305]

[0306] In some embodiments of Formula Ib, CP 1b A fibrin-binding peptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0307]

[0308] Also provided herein are compounds of formula II:

[0309] [M 2 ] r –[C 2 ] s –[CP 2 ]–[R 2 ] t (II)

[0310] or a pharmaceutically acceptable salt thereof,

[0311] Among them, each M 2 independently copper-64 or gallium-68;

[0312] Each C 2 independently a chelating moiety;

[0313] CP 2 It is a fibrin-binding peptide;

[0314] Each R 2 independently organic non-chelating moieties;

[0315] r is an integer selected from 0-5;

[0316] s is an integer selected from 0-5; and

[0317] t is an integer selected from 0-5.

[0318] In some embodiments of Formula II, each M 2 is copper-64. In some embodiments of Formula II, each M 2 It is gallium-68.

[0319] In some embodiments of Formula II, each C 2 Independently selected from the group consisting of:

[0320]

[0321] In some embodiments of Formula II, C 2 For NODAGA:

[0322] In some embodiments of Formula II, each C 2 Independently selected from the group consisting of:

[0323]

[0324] In some embodiments of Formula II, CP 2 is a fibrin-binding peptide comprising a sequence having at least 80% sequence identity to the polypeptide of SEQ ID NO: 16:

[0325] –y*–x 5 –C–Hyp–Y(3-Cl)–X 6 –L–C–X 7 –I–X 8 – (SEQ ID NO: 16)

[0326] Among them, X 5, X 6 , X 7 and X 8 Each is independently any amino acid; y* is L-tyrosine or D-tyrosine. 2 is a fibrin-binding peptide comprising a sequence having at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to the polypeptide of SEQ ID NO: 16. In some embodiments, CP 2 The fibrin-binding peptide is SEQ ID NO: 16 (ie, has 100% sequence identity to SEQ ID NO: 16).

[0327] In some embodiments, y* is L-tyrosine. In some embodiments, y* is D-tyrosine.

[0328] In some embodiments, X 5 , X 6 , X 7 and X 8 Each is independently selected from naturally occurring amino acids. For example, X 5 , X 6 , X 7 and X 8 Each is independently selected from Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp and Tyr. In some embodiments, X 5 , X 6 , X 7 and X 8 Each is independently selected from the D-configuration of naturally occurring amino acids. 5 , X 6 , X 7 , X 8 Each is independently selected from D-Ala, D-Cys, D-Asp, D-Glu, D-Phe, D-His, D-Ile, D-Lys, D-Leu, D-Met, D-Asn, D-Pro, D-Gln, D-Arg, D-Ser, D-Thr, D-Val, D-Trp and D-Tyr. In some embodiments, X 5 In some embodiments, X 5 In some embodiments, X 6 In some embodiments, X 6 In some embodiments, X 6 In some embodiments, X 7In some embodiments, X 7 In some embodiments, X 8 In some embodiments, X 8 In some embodiments, X 8 In some embodiments, X 8 It is D-Leu.

[0329] In some embodiments, X 5 , X 6 , X 7 and X 8 Each is independently selected from non-naturally occurring amino acids. 5 , X 6 , X 7 and X 8 Each is independently selected from Hyp, D-Hyp, Tyr-3-Cl and D-Tyr-3-Cl.

[0330] In some embodiments of Formula II, CP 2 A fibrin-binding peptide comprising a polypeptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0331]

[0332] In some embodiments of Formula II, CP 2 A fibrin-binding peptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0333]

[0334]

[0335] In some embodiments of Formula II, each R 2 Independently selected from –N[(CH2) a OR a ]2. –NH(CH2) a OR a 、–NH(CH2) a OH, –NH(CH2)CH3, –N[(CH2) a CH3], –NH(CF2) a CF3, OR a and

[0336] Among them, each R a are independently (C1-C6)alkyl; and

[0337] a is an integer selected from 0-5.

[0338] In some embodiments, r is 1. In some embodiments, r is 2. In some embodiments, s is 1. In some embodiments, s is 2. In some embodiments, t is 1. In some embodiments, t is 2. In some embodiments, r, s, and t are each 1. In some embodiments, r, s, and t are each 2.

[0339] Also provided herein are compounds of formula III:

[0340] [R 3 ] u –[CP 3 ]–[C 3 ] v –[M 3 ] w (III)

[0341] or a pharmaceutically acceptable salt thereof,

[0342] Among them, each M 3 independently copper-64 or gallium-68;

[0343] Each C 3 is the chelating part;

[0344] CP 3 It is a fibrin-binding peptide;

[0345] Each R 3 independently organic non-chelating moieties;

[0346] u is an integer selected from 0-5;

[0347] v is an integer selected from 0-5; and

[0348] w is an integer selected from 0-5.

[0349] In some embodiments of Formula III, each M 3 is copper-64. In some embodiments of Formula III, each M 3 It is gallium-68.

[0350] In some embodiments of Formula III, each C 3 Independently selected from the group consisting of:

[0351]

[0352] In some embodiments of Formula III, C 3 For NODAGA:

[0353] In some embodiments of Formula III, each C 3 Independently selected from the group consisting of:

[0354]

[0355]

[0356] In some embodiments of Formula III, CP 3 is a fibrin-binding peptide comprising a sequence having at least 80% sequence identity to the polypeptide of SEQ ID NO: 17:

[0357] –y*–x 9 –C–Hyp–Y(3-Cl)–X 10 –L–C–X 11 –I–X 12 – (SEQ ID NO: 17)

[0358] Among them, X 9 , X 10 , X 11 and X 12 Each is independently any amino acid; y* is L-tyrosine or D-tyrosine. 3 is a fibrin-binding peptide comprising a sequence having at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to the polypeptide of SEQ ID NO: 17. In some embodiments, CP 3 The fibrin-binding peptide is SEQ ID NO: 17 (ie, has 100% sequence identity to SEQ ID NO: 17).

[0359] In some embodiments, y* is L-tyrosine. In some embodiments, y* is D-tyrosine.

[0360] In some embodiments, X 9 , X 10 , X 11 and X 12 Each is independently selected from naturally occurring amino acids. For example, X 9 , X 10 , X 11 and X 12Each is independently selected from Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp and Tyr. In some embodiments, X 9 , X 10 , X 11 and X 12 Each is independently selected from the D-configuration of naturally occurring amino acids. 9 , X 10 , X 11 and X 12 Each is independently selected from D-Ala, D-Cys, D-Asp, D-Glu, D-Phe, D-His, D-Ile, D-Lys, D-Leu, D-Met, D-Asn, D-Pro, D-Gln, D-Arg, D-Ser, D-Thr, D-Val, D-Trp and D-Tyr. In some embodiments, X 9 In some embodiments, X 9 In some embodiments, X 10 In some embodiments, X 10 In some embodiments, X 10 In some embodiments, X 11 In some embodiments, X 11 In some embodiments, X 12 In some embodiments, X 12 In some embodiments, X 12 In some embodiments, X 12 It is D-Leu.

[0361] In some embodiments, X 9 , X 10 , X 11 and X 12 Each is independently selected from non-naturally occurring amino acids. 9 , X 10 , X 11 and X 12 Each is independently selected from Hyp, D-Hyp, Tyr-3-Cl and D-Tyr-3-Cl.

[0362] In some embodiments of Formula III, CP 3A fibrin-binding peptide comprising a polypeptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0363]

[0364] In some embodiments of Formula III, CP 3 A fibrin-binding peptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0365]

[0366]

[0367] In some embodiments of Formula III, each R 3 Independently selected from –N[(CH2) b OR b ]2. –NH(CH2) b OR b 、–NH(CH2) b OH, –NH(CH2)CH3, –N[(CH2) b CH3], –NH(CF2) b CF3, OR b and

[0368] Among them, each R b are independently (C1-C6)alkyl; and

[0369] b is an integer selected from 0-5.

[0370] In some embodiments, u is 1. In some embodiments, u is 2. In some embodiments, v is 1. In some embodiments, v is 2. In some embodiments, w is 1. In some embodiments, w is 2. In some embodiments, u, v, and w are each 1. In some embodiments, u, v, and w are each 2.

[0371] Also provided herein are compounds of formula IV:

[0372]

[0373] or a pharmaceutically acceptable salt thereof,

[0374] Among them, R 4 It is a radioactive isotope;

[0375] C 4 is a chelating moiety selected from the group consisting of:

[0376]

[0377]

[0378]

[0379]

[0380] CP 4 It is a fibrin-binding peptide;

[0381] AA is the N-terminal amino acid of the fibrin-binding peptide;

[0382] L 4 For the joint;

[0383] y is an integer selected from 0 and 1; and

[0384] z is an integer selected from 0 and 1.

[0385] In some embodiments of Formula IV, R 4 is a radioisotope selected from therapeutic radioisotopes and radioisotopes detectable using nuclear imaging techniques. 4 Selected from fluorine-18, aluminum fluoride (Al 18 F), scandium-43, scandium-44, scandium-47, manganese-51, manganese-52, copper-60, copper-61, copper-62, copper-64, copper-67, gallium-67, gallium-68, yttrium-86, zirconium-89, technetium-99m, yttrium-90, indium-111, iodine-123, iodine-124, iodine-125, iodine-131, terbium-149, terbium-152, samarium-153, terbium-155, terbium-161, holmium-166, lutetium-177, rhenium-188, lead-203, astatine-211, lead-212, bismuth-213, radium-223, actinium-225, and thorium-227.

[0386] In some embodiments of Formula IV, R 4is a radioisotope that is a therapeutic radioisotope. In some embodiments, the therapeutic radioisotope is selected from scandium-47, copper-67, yttrium-90, iodine-131, samarium-153, terbium-161, holmium-166, lutetium-177, rhenium-188, astatine-211, lead-212, bismuth-213, radium-223, actinium-225, and thorium-227. In some embodiments, the therapeutic isotope is selected from yttrium-90, lutetium-177, and actinium-225. In some embodiments, the therapeutic isotope is yttrium-90. In some embodiments, the therapeutic radioisotope is lutetium-177. In some embodiments, the therapeutic radioisotope is actinium-225.

[0387] In some embodiments of Formula IV, R 4 is a radioactive isotope that can be detected using nuclear imaging techniques. In some embodiments, the radioactive isotope is a positron emitting isotope. In some embodiments, the positron emitting isotope is selected from fluorine-18, aluminum fluoride (Al 18 In some embodiments, the positron emitting isotope is selected from fluorine-18, copper-64 and gallium-68. In some embodiments, the positron emitting isotope is fluorine-18. In some embodiments, the positron emitting isotope is copper-64. In some embodiments, the positron emitting isotope is gallium-68. In some embodiments, the positron emitting isotope is copper-64. In some embodiments, the positron emitting isotope is gallium-68. In some embodiments, the radioactive isotope is a radioactive isotope suitable for SPECT imaging. In some embodiments, the radioisotope suitable for SPECT imaging is selected from gallium-67, technetium-99m, indium-111, iodine-123, iodine-125, terbium-155, and lead-203.

[0388] In some embodiments of Formula IV, C 4 Select from the following group:

[0389]

[0390]

[0391] In some embodiments of Formula IV, C 4 For NODAGA:

[0392] In some embodiments of Formula IV, C 4 Select from the following group:

[0393]

[0394] In some embodiments, C 4 Select from the following group:

[0395]

[0396]

[0397] In some embodiments of Formula IV, AA-CP 4 is a fibrin-binding peptide comprising a sequence having at least 80% sequence identity to the polypeptide of SEQ ID NO: 1:

[0398] –y*–x 1 –C–Hyp–Y(3-Cl)–X 2 –L–C–X 3 –I–X 4 – (SEQ ID NO: 1)

[0399] Among them, X 1 , X 2 , X 3 and X 4 Each is independently any amino acid; y* is L-tyrosine or D-tyrosine; AA represents the N-terminal amino acid. For example, AA-CP 4 is a fibrin-binding peptide comprising a sequence having at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to the polypeptide of SEQ ID NO: 1. In some embodiments, AA-CP 4 The polypeptide is SEQ ID NO: 1 (ie it has 100% sequence identity).

[0400] In some embodiments, y* is L-tyrosine. In some embodiments, y* is D-tyrosine.

[0401] In some embodiments, X 1 , X 2 , X 3 and X 4 Each is independently selected from naturally occurring amino acids. For example, X 1 , X 2 , X 3 and X 4 Each is independently selected from Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp and Tyr. In some embodiments, X 1 , X 2 , X3 and X 4 Each is independently selected from the D-configuration of naturally occurring amino acids. 1 , X 2 , X 3 , X 4 Each is independently selected from D-Ala, D-Cys, D-Asp, D-Glu, D-Phe, D-His, D-Ile, D-Lys, D-Leu, D-Met, D-Asn, D-Pro, D-Gln, D-Arg, D-Ser, D-Thr, D-Val, D-Trp and D-Tyr. In some embodiments, X 1 In some embodiments, X 1 In some embodiments, X 2 In some embodiments, X 2 In some embodiments, X 2 In some embodiments, X 3 In some embodiments, X 3 In some embodiments, X 4 In some embodiments, X 4 In some embodiments, X 4 In some embodiments, X 4 It is D-Leu.

[0402] In some embodiments, X 1 , X 2 , X 3 and X 4 Each is independently selected from non-naturally occurring amino acids. 1 , X 2 , X 3 and X 4 Each is independently selected from Hyp, D-Hyp, Tyr-3-Cl and D-Tyr-3-Cl.

[0403] In some embodiments of Formula IV, AA-CP 4 A fibrin-binding peptide comprising a polypeptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0404]

[0405]

[0406] In some embodiments of Formula IV, AA-CP 4 A fibrin binding peptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0407]

[0408] In some embodiments of Formula IV, AA-CP 4 A fibrin binding peptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0409]

[0410]

[0411] In some embodiments of Formula IV, AA-CP 4 A fibrin binding peptide having at least 80% (e.g., at least 85%, at least 90%, at least 92%, at least 95%, at least 98%, at least 99% or 100%) sequence identity with a polypeptide selected from the group consisting of:

[0412]

[0413] In some embodiments of Formula IV, [(L 4 ) z- The moiety (AA)] represents a fibrin-binding peptide CP optionally modified with a linker 4 wherein AA is the N-terminal amino acid of the fibrin binding peptide, L 4 is an optional connector. 4 It may be a peptide comprising an N-terminus that can bind to an amino acid (e.g., a fibrin binding peptide CP 4 any compound with a functional group that forms a covalent bond with the N-terminus of

[0414] In some embodiments, L 4 is selected from C1-C6 alkyl, 6-10 membered aryl, 5-10 membered heteroaryl, C1-C6 alkyl-C(O)-, 6-10 membered aryl-C(O)- and 5-10 membered heteroaryl-C(O)-. In some embodiments, L 4 is a 5-6 membered heteroaryl. 4In some embodiments, L 4 is 5-6 membered heteroaryl-C(O)-. In some embodiments, L 4 is (pyridinyl)-C(O)-. In some embodiments, L 4 It is (pyridin-3-yl)-C(O)-.

[0415] In some embodiments of the compound of Formula IV, [(C 4 ) y -(R 4 ))]Partially through L 4 Group and [(L 4 ) z -(AA)] moiety, wherein the chelating moiety C 4 With L 4 In some embodiments, [(C 4 ) y -(R 4 ))]Partially through L 4 Group and [(L 4 ) z -(AA)] part, where the radioisotope R 4 With L 4 In some embodiments, [(C 4 ) y -(R 4 ))] part through the AA group and [(L 4 ) z -(AA)] part, where the radioisotope R 4 In some embodiments, [(C 4 ) y -(R 4 ))] part through the AA group and [(L 4 ) z -(AA)] moiety, wherein the chelating moiety C 4 Combined with AA group.

[0416] In some embodiments of Formula IV, y is 0. In some embodiments, y is 1.

[0417] In some embodiments of Formula IV, z is 0. In some embodiments, z is 1.

[0418] In some embodiments of Formula IV, y is 0 and z is 0. In some embodiments, y is 0 and z is 1. In some embodiments, y is 1 and z is 0. In some embodiments, y is 1 and z is 1.

[0419] In some embodiments, the compound of formula IV is a compound of formula IVa:

[0420]

[0421] or a pharmaceutically acceptable salt thereof, wherein R 4 is capable of being chelated by part C 4 Chelated radioisotopes.

[0422] In some embodiments, R 4 Selected from aluminum fluoride (Al 18 F), scandium-43, scandium-44, scandium-47, manganese-51, manganese-52, copper-60, copper-61, copper-62, copper-64, copper-67, gallium-67, gallium-68, yttrium-86, zirconium-89, technetium-99m, yttrium-90, indium-111, terbium-149, terbium-152, samarium-153, terbium-155, terbium-161, holmium-166, lutetium-177, rhenium-188, lead-203, lead-212, bismuth-213, radium-223, actinium-225, and thorium-227.

[0423] In some embodiments of the compound of Formula IVa, z is 0 and [(C 4 )-(R 4 )] part through the chelating part C 4 In some embodiments, the chelating moiety C 4 Forms an amide bond with the AA group.

[0424] In some embodiments of the compound of Formula IVa, z is 1 and [(C 4 )-(R 4 )] Partially through L 4 Group and [(L 4 )-(AA)] part, wherein the chelating part C 4 With L 4 Group bonding.

[0425] In some embodiments, the compound of formula IV is a compound of formula IVb:

[0426]

[0427] or a pharmaceutically acceptable salt thereof, wherein R 4 is able to be connected with the connector L 4 , the N-terminal amino acid of the fibrin binding peptide AA, or a radioactive isotope covalently bound to both.

[0428] In some embodiments, R 4 Selected from fluorine-18, iodine-123, iodine-124, iodine-125, iodine-131 and astatine-211.

[0429] In some embodiments of the compound of Formula IVb, z is 0 and [R 4 ] part is directly combined with the [AA] part.

[0430] In some embodiments of the compound of Formula IVb, z is 1 and [R 4 ]Partially through L 4 Group and [(L 4 )-(AA)] part. In some embodiments, R 4 Part with L 4 In some embodiments, R 4 It is fluorine-18.

[0431] In some embodiments, the compound of formula IV is a compound of formula IVc:

[0432]

[0433] or a pharmaceutically acceptable salt thereof, wherein [R 4 ]Partially through the L 4 The covalent bond of the group and [(L 4 )-(AA)] part, where R 4 To be able to connect with the connector L 4 , the N-terminal amino acid of the fibrin binding peptide AA, or a radioactive isotope covalently bound to both.

[0434] In some embodiments, R 4 is selected from fluorine-18, iodine-123, iodine-124, iodine-125, iodine-131, and astatine-211. 4 It is fluorine-18.

[0435] In some embodiments, the compound of formula IV is a compound of formula IVd:

[0436]

[0437] or a pharmaceutically acceptable salt thereof, wherein [R 4 ] part is directly combined with the [AA] part, where R 4 To be able to connect with the connector L 4 , the N-terminal amino acid of the fibrin binding peptide AA, or a radioactive isotope covalently bound to both.

[0438] In some embodiments, R 4 Selected from fluorine-18, iodine-123, iodine-124, iodine-125, iodine-131 and astatine-211.

[0439] In some embodiments of Formula IV, the compound is selected from the group consisting of:

[0440]

[0441]

[0442]

[0443] or a pharmaceutically acceptable salt thereof.

[0444] In some embodiments of Formula IV, the compound is selected from the group consisting of:

[0445]

[0446]

[0447]

[0448] or a pharmaceutically acceptable salt thereof.

[0449] In some embodiments of Formula IV, the compound is:

[0450]

[0451] or a pharmaceutically acceptable salt thereof.

[0452] In some embodiments of Formula IV, the compound is:

[0453]

[0454] or a pharmaceutically acceptable salt thereof.

[0455] In some embodiments of Formula IV, the compound is selected from the group consisting of:

[0456]

[0457] as well as

[0458]

[0459] or a pharmaceutically acceptable salt thereof.

[0460] In some embodiments of Formula IV, the compound is selected from the group consisting of:

[0461]

[0462]

[0463] The present disclosure also provides compounds of formula V:

[0464]

[0465] or a pharmaceutically acceptable salt thereof, wherein C 4 , L 4 ,AA,CP 4 , y and z refer to the description of the compound of formula IV in this disclosure.

[0466] In some embodiments, the compound of formula V is a compound selected from the group consisting of:

[0467]

[0468]

[0469] Pharmaceutically acceptable derivatives and compositions

[0470] In some embodiments, the compounds of the present disclosure can be formulated into pharmaceutical compositions. In some embodiments, the pharmaceutical composition comprises a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises a compound of formula II or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises a compound of formula III or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises a compound of formula IV or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0471] As used herein, a compound may include a pharmaceutically acceptable derivative thereof. "Pharmaceutically acceptable" means that a compound or composition can be administered to an animal without unacceptable side effects. "Pharmaceutically acceptable derivative" refers to any pharmaceutically acceptable salt, ester, or salt of an ester of a compound of the present disclosure, which is capable of providing (directly or indirectly) a compound or an active metabolite or residue thereof when administered to a recipient.

[0472] Other derivatives are those that increase the bioavailability of the compound when these compounds are administered to a mammal (e.g., by making an orally administered compound more readily absorbed into the blood) or that enhance the delivery of the parent compound to a biological compartment (e.g., the brain or the lymphatic system) thereby increasing exposure relative to the parent substance.

[0473] Pharmaceutically acceptable salts of the compounds of the present disclosure include counterions derived from pharmaceutically acceptable inorganic and organic acids and bases known in the art, for example, alkali metal and alkaline earth metal cations; sodium; primary, secondary and tertiary amines such as ethanolamine, diethanolamine, morpholine, glucosamine, N,N-dimethylglucamine, N-methylglucamine; and amino acids such as lysine, arginine and ornithine.

[0474] Conventional excipients such as adhesives, fillers, acceptable wetting agents, tableting lubricants and disintegrants can be used in tablets and capsules for oral administration. Liquid preparations for oral administration can be in the form of solutions, emulsions, aqueous or oily suspensions and syrups. Alternatively, oral preparations can be in the form of dry powders, which can be reconstituted with water or another suitable liquid carrier before use. Additional additives such as suspending agents or emulsifiers, non-aqueous carriers (including edible oils), preservatives, flavorings and coloring agents can be added to liquid preparations. Parenteral dosage forms can be prepared by dissolving a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof in a suitable liquid carrier and filtering and sterilizing the solution before filling and sealing a suitable vial or ampoule. These are just a few examples of many suitable methods for preparing dosage forms well known in the art.

[0475] The compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof can be formulated into a pharmaceutical composition using techniques well known to those skilled in the art. Suitable pharmaceutically acceptable carriers are known in the art, except those mentioned herein; for example, see Remington, The Science and Practice of Pharmacy, 20th edition, 2000, Lippincott Williams & Wilkins, (author: Gennaro, AR et al.).

[0476] Pharmaceutical compositions as described herein can be administered by any route, including oral and parenteral administration. Parenteral administration includes, but is not limited to, subcutaneous, intravenous, intra-arterial, interstitial, intrathecal, and intracavitary administration. When administered intravenously, the pharmaceutical composition can be injected, administered as two or more closures separated in time, or as a constant or nonlinear flow infusion. Therefore, the compositions of the present disclosure can be formulated for any route of administration.

[0477] In some embodiments, the pharmaceutical composition for intravenous administration is a solution in a sterile isotonic aqueous buffer. In some embodiments, the composition may also include one or more solubilizers, stabilizers, and local anesthetics (e.g., lidocaine) for alleviating pain at the injection site. In some embodiments, the composition for intravenous administration includes sucrose (e.g., 80 mmoles). In some embodiments, these components will be provided separately (e.g., in a test kit) or mixed together in a unit dosage form (e.g., as a dry lyophilized powder or anhydrous concentrate). The composition may be stored in a sealed container such as, for example, an ampoule or a pouch, and the amount of the active agent is expressed in active units. When the composition is administered by infusion, it may be distributed with an infusion bottle containing sterile pharmaceutical grade "water for injection", saline or other suitable intravenous infusions. In the case of administering the composition by injection, an ampoule of sterile water for injection or saline may be provided as a component of the test kit so that the components may be mixed before administration.

[0478] The pharmaceutical composition of the present disclosure containing a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof may further contain one or more other ingredients. Examples of such ingredients include, but are not limited to, pH regulators, stabilizers, detergents and isotonic agents. In some embodiments, the pharmaceutical composition contains one or more of acetic acid, sodium acetate, sodium hydroxide, gentisic acid, ascorbic acid, diethylenetriamine pentaacetic acid (DPTA) and sodium chloride. In some embodiments, the pharmaceutical composition contains water for injection.

[0479] In some embodiments, the pharmaceutical composition comprising a compound of Formula I, II, III or IV or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient comprises a free radical scavenger. Free radical scavengers can be used to prevent radiolysis. Radiolysis is a process in which other active substances such as superoxide, hydrogen peroxide, hydrogen radicals, ozone and hydroxyl radicals are formed by the ionization of oxygen or water molecules induced by radionuclides. These active substances can further damage DNA and other cell structures. In some embodiments, the free radical scavenger is an antioxidant selected from carnosic acid, green tea extract, apigenin, diosmin, rosmarinic acid, lipoic acid, beta-carotene, L-ascorbic acid (vitamin C), N-acetylcysteine ​​(NAC), δ-tocopherol, rutin, amifostine, resveratrol, gentisic acid and gallic acid. In some embodiments, the free radical scavenger is an antioxidant selected from gallic acid, L-ascorbic acid and N-acetylcysteine ​​(NAC).

[0480] In some embodiments, the compositions of the present disclosure are administered to patients in the form of injectable compositions. In some embodiments, the method of administering the compound is parenteral, i.e., intravenous, intra-arterial, intrathecal, interstitial or intracavitary. The pharmaceutical compositions of the present invention can be administered to animals, including humans, in a manner similar to other diagnostic or therapeutic agents. The dosage and mode of administration will depend on a variety of factors, including age, weight, sex, patient condition and genetic factors, and ultimately the imaging as described herein will be determined by medical personnel after experimental determination of different dosages.

[0481] Fibrin imaging methods

[0482] The compounds and compositions of the present disclosure can be used for fibrin imaging. In some embodiments, fibrin is present in a tumor. In some embodiments, the tumor is cancerous. In some embodiments, fibrin is present in a thrombus. In some embodiments, fibrin is associated with neuroinflammation. In some embodiments, neuroinflammation is associated with Alzheimer's disease, multiple sclerosis, or traumatic brain injury.

[0483] In some embodiments, a method of imaging fibrin in a mammal comprises administering to the mammal an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof; acquiring an image of the fibrin in the mammal using nuclear imaging techniques and acquiring an anatomical image of the mammal using magnetic resonance imaging (MRI); and superimposing the images to locate the fibrin within the anatomical image of the mammal.

[0484] In some embodiments, a method for imaging fibrin in a mammal comprises administering to the mammal an effective amount of a pharmaceutical composition of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient; acquiring an image of the fibrin in the mammal using nuclear imaging techniques and acquiring an anatomical image of the mammal using magnetic resonance imaging; and superimposing the images to locate the fibrin within the anatomical image of the mammal.

[0485] In some embodiments of the imaging method of fibrin, the fibrin image of the mammal using nuclear imaging technology and the image of the mammal using magnetic resonance imaging are acquired simultaneously. In some embodiments of the imaging method of fibrin, the fibrin image of the mammal using nuclear imaging technology is acquired first, and then the image of the mammal using magnetic resonance imaging is acquired. In some embodiments of the imaging method of fibrin, the image of the mammal using magnetic resonance imaging is acquired first, and then the fibrin image of the mammal using nuclear imaging technology is acquired.

[0486] In some embodiments of the methods of imaging fibrin, the nuclear imaging technique is single photon emission computed tomography (SPECT).

[0487] In some embodiments of the methods of imaging fibrin, the nuclear imaging technique is positron emission tomography (PET). In some embodiments of the methods of imaging fibrin, the nuclear imaging technique is a combination of positron emission tomography and computed tomography (PET-CT).

[0488] In some embodiments of the imaging method of fibrin, the mammal is a human. In some embodiments of the imaging method of fibrin, the mammal is a rat. In some embodiments of the imaging method of fibrin, the mammal is a dog.

[0489] In some embodiments of the method of imaging fibrin, the method further comprises administering to the mammal an effective amount of a second compound or composition. In some embodiments, the second compound or composition is not directed against fibrin.

[0490] In some embodiments, the second compound or a composition comprising the second compound is a second imaging agent. In some embodiments, the second imaging agent comprises an MRI imaging agent. In some embodiments, the second imaging agent is an MRI imaging agent, for example, gadoteridol, gadopentetate, gadobenzoic acid, gadoxetic acid, gadodiamine, gadovesamine, and gadofosveset; or a CT imaging agent selected from iopamidol, iohexol, ioxilan, iopromide, iodixanol, ioxaglate, metrizoate, and diatrizoate.

[0491] In some embodiments, the second compound or a composition comprising the second compound contains a therapeutic radioisotope. In some embodiments, the therapeutic radioisotope is selected from scandium-47, copper-67, yttrium-90, iodine-131, samarium-153, terbium-161, holmium-166, lutetium-177, rhenium-188, astatine-211, lead-212, bismuth-213, radium-223, actinium-225, and thorium-227. In some embodiments, the therapeutic isotope is selected from yttrium-90, lutetium-177, and actinium-225. In some embodiments, the therapeutic isotope is yttrium-90. In some embodiments, the therapeutic isotope is lutetium-177. In some embodiments, the therapeutic isotope is actinium-225.

[0492] Image Overlay

[0493] The superposition of images can be accomplished by various means known in the art. For example, see U.S. Patent Nos. 7,412,279; 7,110616; 6,898,331; 6,549,798; and 5,672,877; Rudd, J. HF. et al., J. Nucl. Med. 2008 49 (6): 871-878; Slomka, PJ et al., J. Nucl. Med. 2009 50: 1621-1630; and Jupp, B. and O'Brien, TJ, Epilepsia 2007 49: 82-89. In some embodiments, the first and second image data sets can be overlapped to determine the presence of fibrin in a mammal. For example, the first and second image data sets can be combined to generate a third data set that includes an image of a fibrin target and an image of an anatomical region where the fibrin is located. The third set of data can indicate the location of fibrin (if present) in a mammal. If desired, the third data set may be displayed on a display device to indicate the location of the immobilized target within the vasculature. The third data set may also indicate the size of the immobilized target within the mammal.

[0494] Treatment

[0495] The present disclosure also provides methods for treating a disease or condition associated with the presence of fibrin. In some embodiments, the disease or condition associated with the presence of fibrin is a cardiovascular disease. In some embodiments, the disease or condition associated with the presence of fibrin is cancer.

[0496] In some embodiments, the method for treating a disease or condition associated with the presence of fibrin in a mammal comprises administering to the mammal an effective amount of a compound of Formula I, Formula II, Formula III or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III or Formula IV, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioisotope. In some embodiments, the method for treating a disease or condition associated with the presence of fibrin in a mammal comprises administering to the mammal a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, wherein the compound contains a therapeutic radioisotope. In some embodiments, the method for treating a disease or condition associated with the presence of fibrin in a mammal comprises administering to the mammal a pharmaceutical composition comprising a compound of Formula II or a pharmaceutically acceptable salt thereof, wherein the compound contains a therapeutic radioisotope. In some embodiments, the method for treating a disease or condition associated with the presence of fibrin in a mammal comprises administering to the mammal a pharmaceutical composition comprising a compound of Formula III or a pharmaceutically acceptable salt thereof, wherein the compound contains a therapeutic radioisotope. In some embodiments, the method for treating a disease or condition related to the presence of fibrin in a mammal comprises administering to a mammal a pharmaceutical composition comprising a compound of formula IV or a pharmaceutically acceptable salt thereof, wherein the compound contains a therapeutic radioisotope. In some embodiments, the disease or condition related to the presence of fibrin is a cardiovascular disease or condition. In some embodiments, the disease or condition related to the presence of fibrin is cancer. In some embodiments, the disease or condition related to the presence of fibrin is a cerebrovascular disease (e.g., cerebral aneurysm, carotid stenosis, spinal stenosis, and stroke). In some embodiments, the mammal is a rat, a mouse, a dog, or a pig. In some embodiments, the mammal is a human.

[0497] The present invention also provides a method for treating cancer in a mammal. In some embodiments, the method comprises administering to a mammal an effective amount of a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioactive isotope. In some embodiments, the method for treating cancer in a mammal comprises administering to a mammal a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, wherein the compound contains a therapeutic radioactive isotope. In some embodiments, the method for treating cancer in a mammal comprises administering to a mammal a pharmaceutical composition comprising a compound of formula II or a pharmaceutically acceptable salt thereof, wherein the compound contains a therapeutic radioactive isotope. In some embodiments, the method for treating cancer in a mammal comprises administering to a mammal a pharmaceutical composition comprising a compound of formula III or a pharmaceutically acceptable salt thereof, wherein the compound contains a therapeutic radioactive isotope. In some embodiments, the method for treating cancer in a mammal comprises administering to a mammal a pharmaceutical composition comprising a compound of formula IV or a pharmaceutically acceptable salt thereof, wherein the compound contains a therapeutic radioactive isotope.

[0498] Examples of cancers that can be treated using the compounds of the present disclosure include, but are not limited to, sarcomas, bone cancer, breast cancer, lung cancer (e.g., non-small cell lung cancer and small cell lung cancer), genitourinary tract cancer, pancreatic cancer, liver cancer, skin cancer, melanoma (e.g., metastatic malignant melanoma, BRAF and HSP90 inhibited melanoma), head and neck cancer, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer anal cancer, gastric cancer, renal cancer (e.g., clear cell carcinoma), prostate cancer (e.g., hormone-refractory prostate adenocarcinoma), testicular cancer, colon cancer, gynecological cancer, uterine cancer, fallopian tube cancer, urothelial cancer (e.g., bladder), endometrial cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's disease, The compounds of the present invention can also be used to treat metastatic cancer.

[0499] In some embodiments, cancers that can be treated using the compounds of the present disclosure include, but are not limited to, solid tumors (e.g., prostate cancer, colon cancer, esophageal cancer, endometrial cancer, ovarian cancer, uterine cancer, kidney cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, sarcoma, bladder cancer), hematological cancers (e.g., lymphomas, leukemias such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), DLBCL, mantle cell lymphoma, non-Hodgkin lymphoma (including follicular lymphoma, including relapsed or refractory NHL and relapsed follicular lymphoma), Hodgkin lymphoma, or multiple myeloma), and combinations of the aforementioned cancers.

[0500] In some embodiments, cancers that may be treated using the compounds of the present disclosure include, but are not limited to, bile duct cancer, cholangiocarcinoma, triple negative breast cancer, rhabdomyosarcoma, small cell lung cancer, leiomyosarcoma, hepatocellular carcinoma, Ewing's sarcoma, brain cancer, brain tumor, astrocytoma, neuroblastoma, neurofibroma, basal cell carcinoma, chondrosarcoma, epithelioid sarcoma, eye cancer, fallopian tube cancer, gastrointestinal cancer, gastrointestinal stromal tumors, hairy cell leukemia, intestinal cancer, islet cell carcinoma, oral cancer, oral cancer, laryngeal cancer, laryngeal cancer, lip cancer, mesothelioma, neck cancer, nasal cancer, eye cancer, eye melanoma, pelvic cancer, rectal cancer, renal cell carcinoma, salivary gland cancer, paranasal sinus cancer, spinal cancer, tongue cancer, renal tubular cancer, urethral cancer, and ureteral cancer.

[0501] Exemplary hematological cancers include lymphomas and leukemias, such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, non-Hodgkin lymphoma (including relapsed or refractory NHL and relapsed follicular lymphoma), Hodgkin lymphoma, myeloproliferative disorders (e.g., primary myelofibrosis (PMF), polycythemia vera (PV), and essential thrombocythemia (ET)), myelodysplastic syndrome (MDS), T-cell acute lymphoblastic lymphoma (T-ALL), and multiple myeloma (MM).

[0502] Exemplary sarcomas include chondrosarcoma, Ewing sarcoma, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxoma, rhabdomyoma, rhabdomyosarcoma, fibroma, lipoma, sarcoma, and teratoma.

[0503] Exemplary lung cancers include non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), bronchogenic carcinoma, squamous cell carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, adenocarcinoma, alveolar (bronchiolar) carcinoma, bronchial adenoma, cartilaginous hamartoma, and mesothelioma.

[0504] Exemplary gastrointestinal cancers include esophageal cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), gastric cancer (carcinoma, lymphoma, leiomyosarcoma), pancreatic cancer (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, hemangioma), small intestine cancer (adenocarcinoma), lymphoma, carcinoid, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), colorectal cancer (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma) and colorectal cancer.

[0505] Exemplary genitourinary tract cancers include renal cancer (adenocarcinoma, Wilms' tumor [Nephroblastoma]), bladder cancer and urethral cancer (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate cancer (adenocarcinoma, sarcoma), and testicular cancer (seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma).

[0506] Exemplary liver cancers include hepatocellular carcinoma, cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma.

[0507] Exemplary bone cancers include, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochondroma (osteocartilaginous exostosis), benign enchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor.

[0508] Exemplary cancers of the nervous system include cancers of the skull (osteomas, hemangiomas, granulomas, xanthomas, osteitis deformans), meninges (meningiomas, meningosarcomas, gliomatosis), brain (astrocytomas, medulloblastomas, gliomas, ependymomas, germ cell tumors (pinealomas), glioblastomas, glioblastomas multiforme, oligodendrogliomas, schwannomas, retinoblastomas, congenital tumors) and spinal cord (neurofibromas, meningiomas, gliomas, sarcomas), as well as neuroblastoma and Lhermitte-Duclos disease.

[0509] Exemplary gynecological cancers include uterine cancer (endometrial cancer), cervical cancer (cervical carcinoma, preneoplastic cervical dysplasia), ovarian cancer (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-theca cell tumor, Sertoli-Leydig cell tumor, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), and fallopian tube (carcinoma).

[0510] Exemplary skin cancers include melanoma, basal cell carcinoma, Merkel cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevus dysplasia, lipoma, hemangioma, dermatofibroma, and keloid. In some embodiments, diseases and indications that can be treated using the compounds of the present disclosure include, but are not limited to, sickle cell disease (e.g., sickle cell anemia), triple negative breast cancer (TNBC), myelodysplastic syndrome, testicular cancer, bile duct cancer, esophageal cancer, and urothelial cancer.

[0511] In some embodiments, the methods of the present disclosure are used to detect, treat, or detect and treat cancers that exhibit high fibrin levels.

[0512] In some embodiments of the treatment methods disclosed herein, a compound of Formula I, Formula II, Formula III or Formula IV or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III or Formula IV or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioisotope, and the method further comprises administering an amino acid solution to the patient. In some embodiments, the amino acid solution is used to prevent nephrotoxicity associated with radionuclide therapy. In some embodiments, the amino acid solution comprises L-lysine, L-arginine, and pharmaceutically acceptable salts and combinations thereof. In some embodiments, the amino acid solution comprises L-lysine and pharmaceutically acceptable salts thereof. In some embodiments, the amino acid solution comprises L-arginine and pharmaceutically acceptable salts thereof. In some embodiments, the amino acid solution comprises a mixture of L-lysine HCl and L-arginine HCl of about 10 g / L-about 40 g / L, for example, about 12 g / L-about 35 g / L, about 16 g / L-about 32 g / L, about 20 g / L-about 30 g / L, or about 22 g / L-about 28 g / L. In some embodiments, the amino acid solution comprises a mixture of L-Lysine HCl and L-Arginine HCl from about 16g / L to about 32g / L. In some embodiments, the amino acid solution comprises L-Lysine HCl from about 8g / L to about 16g / L and L-Arginine HCl from about 8g / L to about 16g / L. In some embodiments, the amino acid solution comprises L-Lysine HCl from about 8g / L to about 16g / L and L-Arginine HCl from about 8g / L to about 16g / L. In some embodiments, the amino acid solution comprises L-Lysine HCl from about 8g / L, about 9g / L, about 10g / L, about 11g / L, about 12g / L, about 13g / L, about 14g / L, about 15g / L or about 16g / L. In some embodiments, the amino acid solution comprises L-Arginine HCl from about 8g / L, about 9g / L, about 10g / L, about 11g / L, about 12g / L, about 13g / L, about 14g / L, about 15g / L or about 16g / L.

[0513] In some embodiments, the amino acid solution is administered intravenously. In some embodiments, the amino acid solution or a combination thereof is administered before, simultaneously with, or after administration of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioisotope.

[0514] In some embodiments, the amino acid solution is applied before administering a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing an effective amount of a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof. In some embodiments, the amino acid solution is applied about 5 minutes to about 60 minutes before administering a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing an effective amount of a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof, for example, 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 50 minutes, about 55 minutes or about 60 minutes before administering a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing an effective amount of a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof. In some embodiments, the amino acid solution is applied about 30 minutes before administering a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof.

[0515] In some embodiments, the amino acid solution is administered while administering a compound of Formula I, Formula II, Formula III or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III or Formula IV, or a pharmaceutically acceptable salt thereof. In some embodiments, the amino acid solution is co-infused with a compound of Formula I, Formula II, Formula III or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III or Formula IV, or a pharmaceutically acceptable salt thereof.

[0516] In some embodiments, the amino acid solution is applied after administering a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing an effective amount of a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof. In some embodiments, the amino acid solution is applied immediately after administering a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing an effective amount of a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof or about 5 minutes to about 60 minutes later, for example, 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 50 minutes, about 55 minutes or about 60 minutes later. In some embodiments, the amino acid solution is applied about 30 minutes after administering a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing an effective amount of a compound of formula I, formula II, formula III or formula IV or a pharmaceutically acceptable salt thereof.

[0517] In some embodiments, the amino acid solution is administered before, simultaneously with, or after administration of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioactive isotope.

[0518] In some embodiments, the amino acid solution is administered prior to and simultaneously with the administration of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioactive isotope.

[0519] In some embodiments, an amino acid solution is administered before and after administration of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioactive isotope.

[0520] In some embodiments, the amino acid solution is administered simultaneously with and after administration of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioactive isotope.

[0521] In some embodiments of the treatment methods disclosed herein, a compound of Formula I, Formula II, Formula III or Formula IV or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III or Formula IV or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioisotope, and the method further comprises administering an antiemetic to the patient. Antiemetics can be used to reduce nausea and vomiting associated with radiotherapy. In some embodiments, the antiemetic is selected from 5-HT3 receptor antagonists, corticosteroids, neurokinin-1 (NK-1) receptor inhibitors, prochlorperazine, metoclopramide and cannabinoids. In some embodiments, the antiemetic is a 5-HT3 receptor antagonist. In some embodiments, the antiemetic is a NK-1 receptor inhibitor.

[0522] In some embodiments, an antiemetic or a combination thereof is administered before, simultaneously with, or after administration of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioisotope. In some embodiments, an antiemetic is administered before administration of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof. In some embodiments, an antiemetic is administered while administering a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof. In some embodiments, an antiemetic is administered after administration of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof.

[0523] In some embodiments, an antiemetic or a combination thereof is administered before, simultaneously with, or after administration of an amino acid solution such as the amino acid solution described in the present disclosure. In some embodiments, an antiemetic is administered before administration of an amino acid solution. In some embodiments, an antiemetic is administered at the same time as an amino acid solution. In some embodiments, an antiemetic is administered after administration of an amino acid solution. The present disclosure also provides methods for detecting and treating diseases or conditions associated with the presence of fibrin in mammals. In some embodiments, the method comprises using a method described in the present disclosure using a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV, or a pharmaceutically acceptable salt thereof, to detect a disease or condition associated with the presence of fibrin in a mammal, wherein the compound or pharmaceutical composition contains a radioactive isotope that can be detected using nuclear imaging techniques. In some embodiments, the method further comprises treating a disease or condition associated with the presence of fibrin in a mammal using a method described in the present disclosure using a compound of Formula I, Formula II, Formula III or Formula IV, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing an effective amount of a compound of Formula I, Formula II, Formula III or Formula IV, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutical composition contains a therapeutic radioactive isotope. In some embodiments, the disease or condition associated with the presence of fibrin is cancer. Example

[0524] The present invention is further illustrated by the following examples, which do not limit the scope of the present invention described in the claims.

[0525] Example 1: Synthesis and characterization of compounds

[0526] Step A – Synthesis of unmodified peptide

[0527] The peptides were synthesized using 1 to 12 batch reactors equipped with 0.1 mmol of commercially available Rink amide resin (-0.38 mmol / g) on ​​an automatic peptide synthesizer "Liberty Blue" (CEM). Standard Fmoc chemistry was used to extend the peptides on the resin. Fmoc was removed with a DMF solution of 20% piperidine and 0.1 M HOBt. Various amino acids were dissolved in DMF to obtain a 0.2 M solution and coupled to the peptide using a DMF solution of 0.5 M diisopropylcarbodiimide and 1.0 M Oxyma (or HOBt).

[0528] After the synthesis of the peptide was completed on the resin, the peptide was filtered and then cleaved from the resin (TFA / TIS / MSA / 2,2-(ethylenedioxy)ethanedithiol / H2O 95:2.5:2.5:2.5:2.5). The fully deprotected peptide solution was precipitated with ether (40 mL). The peptide solid was separated after centrifugation and then cyclized in a 1:1 mixture of DMSO / 40mM NH4Oac, pH5. The cyclization was monitored by LC-MS (24 hours). The cyclic peptide was purified in 23 minutes by reverse phase preparative HPLC on a C-5 column using a gradient of 5% mobile phase A (0.1% TFA in water) to 60% mobile phase B (0.1% TFA in acetonitrile). The pure peptide fractions were combined and lyophilized to obtain the final peptide portion.

[0529] Step B – Synthesis of N-terminally modified peptides

[0530] Afterwards, the N-terminus of each peptide was modified to: The prepared compounds (eg, N-terminally modified peptides) are shown in Tables 1 and 1a, wherein F is fluorine (Table 1) or fluorine-18 (Table 1a). Unless otherwise indicated, all compounds have a C-terminal amide.

[0531] Table 1.

[0532]

[0533]

[0534] Table 1a.

[0535]

[0536]

[0537]

[0538] Example 2: Synthesis of PFP peptide

[0539] Unmodified peptide was prepared according to the procedure in Example 1 Step A. 6-fluoronicotinic acid-PFP (0.10 mmol) was coupled to the N-terminus of the peptide (0.02 mmol) in DMF (0.50 mL) overnight at 40° C. The reaction was monitored by LCMS. The crude compound obtained was purified by a gradient of 5% mobile phase A (0.1% TFA in water) to 60% mobile phase B (0.1% TFA in acetonitrile) on a reverse phase preparative HPLC C-5 column (22.5×250 mm) for more than 23 minutes, and the combined fractions were lyophilized to obtain the pure compound.

[0540] Example 3: Determination of binding to immobilized fibrin and soluble fibrin fragment DD(E) using fluorescence polarization (FP) assay

[0541] DD(E) was prepared using a published method involving partial plasmin digestion of purified fibrin gels followed by size exclusion chromatography purification and purity assessment by SDS-PAGE. Peptides modified at the N-terminus with tetramethylrhodamine (TRITC) dye were conjugated to DD(E). Fluorescent peptides directly bound to DD(E) were measured by fluorescence polarization (FP) assay. The anisotropy (r) of fluorescently labeled peptides bound to DD(E) (0-20 μM) was measured in Tris-buffered saline (50 mM Tris, 100 mM NaCl, and 2 mM CaCl2) at fixed compound concentrations. obs ), the data were fitted to a single-site model (Eq. 1) to obtain the dissociation constant (K) of the DD(E)·(fluorescent peptide) complex. d ).

[0542]

[0543] The displacement of the fluorescent peptide by the non-fluorescent compound was measured to determine the inhibition constant (K i ). In the presence of an inhibitor, the apparent dissociation constant (K d app) was determined using Equation 2. The inhibition constant Ki and K d app is the relationship of Equation 2, where K d is the true dissociation constant of the fluorescent compound measured in the absence of inhibitor.

[0544]

[0545] Binding of unlabeled peptide to DD(E) was measured by a compound displacement assay, in which the DD(E) / Fl·pep complex formed at fixed concentrations of DD(E) and Fl·pep was titrated with increasing concentrations of unlabeled compound. The compound was assayed by mixing DD(E) (4.0 μM, 10 μL) and TRITC (0.1 μM, 10 μL) with increasing concentrations of competing peptide (0.4-100 μM, 20 μL) in TBS·Ca with 2% DMSO. Fluorescence polarization of increasing concentrations of samples (10 μL, triplicate wells) was measured in a 384-well microplate (Corning black plate) using a Tecan Infinite 200 Pro fluorescence microplate reader equipped with a fluorescence polarization filter, a 535 nm excitation filter, and a 590 nm fluorescence emission filter. The average polarization (mP units) readings of each dilution of the target compound were converted to anisotropy (r) using Equation 3.

[0546]

[0547] The data defines a displacement curve (r obs relative to [peptide]), and the inhibition binding constant K i Determined by least squares fitting of the data (K i ±10% estimated uncertainty). K of Example 1 Compound i See Table 2 for values.

[0548] Table 2.

[0549]

[0550]

[0551] result

[0552] All compounds from series A and C were screened for affinity to the soluble fibrin fragment DD(E) using fluorescence polarization. The compounds were compared to EP-2104R, a reference compound with demonstrated fibrin affinity ( Figure 1 and Figure 2 Of the 14 compounds tested, six (compounds 7, 6, 4, 2, 5, and 3) were identified as having submicromolar affinity (K) for fibrin as assessed by the DD(E) assay. d <0.6 μM).

[0553] Example 4: Determination of metabolic stability using rat serum test

[0554] Six compounds identified in Example 3 (compounds 2, 3, 4, 5, 6 and 7) were tested for stability up to 4 hours in rat plasma at 37°C. The compounds were dissolved in DMSO to make a stock solution (0.6mM). Plasma (995μL) was spiked with a peptide stock solution (5μL). Incubated at a test compound concentration of 3μM, the final DMSO concentration was 2.5%. The spiked plasma samples were incubated at 37°C for 4 hours. Aliquots (300μL) were taken out at 0, 0.5, 1, 2 and 4 hours, and the reaction was terminated by adding 600μL of cold acetonitrile containing 0.1% formic acid and carbutamide (25μg / mL) as an internal standard. At the same time, a plasma sample (995μL) containing a mixture of benfluamine, propranolol and nortriptyline (40μM DMSO solution) control compounds was also incubated with each batch of test compounds and terminated in a similar manner. In addition, matrix blanks were prepared by spiking internal standards containing acetonitrile into plasma samples without any analyte or control compound. Aliquots were centrifuged at 5000 × g for 5 min for protein precipitation. After centrifugation, the concentration of target compounds in the supernatant was quantified by LCMS. The corresponding [(M+2) / 2] + To confirm the presence of the peptide. The peak area of ​​each detected compound was measured to determine the remaining % of each compound, and the results were compared with standards of pure compounds and compounds immediately isolated from plasma (t=0 samples). All test samples were analyzed in triplicate. LCMS measurements were performed using a gradient elution system (5-95%) consisting of water, acetonitrile and 0.1% TFA at a flow rate of 1 mL / min using a Phenomenex-C18 column (3 μm particle size, 4.6 mm×100 mm).

[0555] result

[0556] At the 2-hour time point, the three compounds were more than 80% intact, with compounds 7 and 6 showing the greatest stability after 4 hours ( Figure 3 ).

[0557] Example 5: Optimization of F-18 radiolabeling conditions and development of analytical methods for compounds

[0558] Synthesis of 6-Chloronicotinic Acid-2,3,5,6-Tetrafluorophenyl Ester (Int-1)

[0559] 6-Chloronicotinic acid (1.0 g, 7.1 mmol), tetrafluorophenol (TFP, 1.18 g, 7.1 mmol) in the presence of N, N'-dicyclohexylcarbodiimide (DCC) (1.40 g, 6.81 mmol) in dioxane (35 mL) was reacted at room temperature for 12 h to obtain 6-Cl-Py-PFP (Int-1) ester. Dicyclohexylurea (DCU) was filtered off and the solvent was removed to obtain a crude product, which was crystallized in hot hexane (1.61 g, 85%). 1 H NMR analysis confirmed the composition of the compound.

[0560] N,N,N-trimethyl-5-((2,3,5,6-tetrafluorophenoxy)carbonyl)pyridine-2-trifluoromethanesulfonamide (Int-2) synthesis

[0561] A steady stream of trimethylamine gas was passed through a mixture of Int-1 (1.0 g, 3.3 mmol) in anhydrous THF (15 mL) at room temperature for 3 hours. The obtained N,N,N-trimethyl-5-((2,3,5,6-tetrafluoro-phenoxy)carbonyl)pyridine-2-ammonium chloride was filtered and then washed with diethyl ether (100 mL) and cold dichloromethane (50 mL). The solid (0.53 g, 1.5 mmol) was suspended in DCM (50 mL) under an argon atmosphere. The solution was filtered and the volatile components were removed under reduced pressure. The residue was washed with diethyl ether (3×50 mL) and dried under vacuum to provide Int-2 (0.65 g, 41.6%). By 1 H NMR analysis confirmed the composition of the compound.

[0562] 18 F-Py-TFP and compounds 18 Radiochemical Synthesis of F-7

[0563] By making the 18 F] fluoride 18 O] water passes through the column, making [ 18 F] Fluoride was captured in Chromafix PS-HCO3 pretreated with 1 ml of 1 M potassium carbonate solution and 20 mL of water. - In the column, 0.4 ml of potassium carbonate and 222 (Kry222) was eluted from the column in a 1:1 solution of acetonitrile and water. 18 F]fluoride. 18 F] fluoride was dried by adding acetonitrile (0.5 mL × 3 times) during evaporation and azeotropic evaporation at 105 ° C in a nitrogen stream. After complete drying, a DMSO solution (0.3 mL) containing Int-2 (0.6 mg, 12.54 μmol) was added to the residue and heated at 90 ° C for 5 minutes. The reaction solution was diluted with acetonitrile: 0.1% TFA solution = 1:1 co-solvent and injected into Alltima C 18A semi-preparative column (250 mm×10 mm, 5 μm) was eluted with 0.1% TFA in acetonitrile solution: 0.1% TFA in water solution = 64:36, with a flow rate of 4 mL / min. 18 The F-PFP ester eluted between 13.0 and 13.5 minutes and was collected in C 18 Sep-Pak PlusCartridge. Release from Sep-Pak using 0.9 ml acetonitrile 18 F-PFP ester. DMSO (0.9 ml) containing TEA and peptide (0.5 mg, 0.36 μmol) was added to the reaction vessel. The mixture was heated at 60 °C for 30 minutes. The reaction was terminated by diluting with 0.9 mL of water. The entire aliquot was injected into the Gemini-NX C 18 Semi-preparative chromatography column (250 mm×10 mm, 5 μm), eluted with 0.1% TFA in acetonitrile solution: 0.1% TFA in water solution = 33:67 at a flow rate of 4 mL / min. 18 F-7 eluted between 12.0 and 12.5 minutes and was collected at C 18 Sep-Pak Plus Cartridge. 1 mL of ethanol was used to obtain the product.

[0564] result:

[0565] F-18 radiolabeling conditions were developed along with analytical methods for the optimized compounds. Figure 4 and Figure 5 A 2-step radiochemical synthesis is described, namely the preparation of 18 F-Py-TFP( Figure 4 ) and compounds 18 F-7( Figure 5 ). The blue trace represents the UV detection trace of the non-radioactive pure compound. The radioactivity detector is located after the UV detector, so there is a shift in the retention time between the UV trace and the radioactivity trace.

[0566] Example 6: Evaluation of specificity by competitive binding with fibrinogen and plasma proteins

[0567] A plate-based assay was developed to directly measure the affinity of compounds for fibrin. Fibrin was immobilized and competition from soluble proteins was measured. Fibrin binding of compounds was determined in the presence or absence of human plasma.

[0568] Fibrin plate preparation

[0569] Human fibrinogen (1 g) was dissolved in 30 mL of TBS buffer (50 mM Tris, 150 mM NaCl, 5 mM sodium citrate, pH 7.4) and dialyzed in a Slyde-a-Lyzer (20000 MWCO, Cassette G2) at room temperature. After changing the buffer twice, the fibrinogen was centrifuged (10 minutes, 2000 × g) to remove undissolved material. The fibrinogen concentration was determined by measuring the absorbance at 280 nm. The reserve fibrinogen solution concentration was 32.1 mg / mL. The concentration was determined by HPLC-MS / MS in a 96-well polystyrene microtiter plate. Fibrin plates with alternating rows of clotted fibrin and empty wells were prepared by polymerizing fibrinogen (100 μL; 2.5 mg / mL) with thrombin (1 U / mL) in TBS·citrate supplemented with 7 mM CaCl2 in the wells. The uncovered plates were dried overnight at 37°C to obtain a thin film, which was adsorbed to the plates, sealed with tape, and stored at -20°C until use. The clottable protein in individual fibrinogen batches was determined by measuring the 280 nm absorbance of the soluble portion of the solution before and after thrombin treatment, which was generally ≥96% (fibrin concentration, 7.56 μM).

[0570] F-18 radioactivity-based fibrin affinity and specificity assay protocol

[0571] From the compound 18 Aliquots (100 μL) of the known activity of F-7 (52 μCi, 10 mL TBS buffer solution) were added to a series of 12 known concentrations (0.01-50 μM) of cold compound 7 TBS buffer solution (550 μL) and mixed thoroughly to prepare 12 stock solutions. Each dilution (100 μL) was added to the coagulated fibrin wells and empty wells in duplicate. Human plasma was used instead of TBS buffer and the same protocol was followed. Afterwards, the prepared plate was covered and incubated at room temperature for 2 hours under constant stirring. Aliquots (90 μL) were carefully moved from the supernatant of each well into a tare test tube. All aliquots from the supernatant and aliquots (90 μL) from various stock solutions were weighed and the activity of each sample was measured using a Cobra 5002 γ counter. All data were attenuation corrected.

[0572] Data analysis

[0573] Using the data obtained from the empty wells, the corresponding fibrin clotted wells, and the stock solution, it was determined that [Compound 7] 总 and [Compound 7] 游离 [Compound 7] 结合 The calculation for each sample is as follows:

[0574] [Compound 7]结合 =[Compound 7] 总 -[Compound 7] 游离

[0575] Use [Compound 7] 结合 and known [fibrin] 总 , calculate [compound 7] 结合 / [Fibrin] 总 Active binding sites (N) and dissociation constant (K d ) by converting [compound 7] 结合 / [Fibrin] 总 For [Compound 7] 游离 Confirm by drawing a graph. From the theoretical equation,

[0576]

[0577] N and K d The variables are adjusted by the solver to minimize the absolute error between the observed and theoretical values ​​( Figure 6 ).

[0578] Compound 18 F-7 binds to human fibrin in TBS buffer and in the presence of human plasma

[0579] The similarity of the results obtained in the presence and absence of plasma proteins suggests that the compound 18 F-7 has a higher binding specificity to fibrin than to fibrinogen ( Figure 7 ).

[0580] Embodiment 7: Having 68 Ga, 64 Cu or Al 18 F Chelating Compounds

[0581] NODAGA Chelating Agents

[0582] ( t Synthesis of Bu)3NODAGA-NHS ester: 4-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclononane-1-yl)-5-(tert-butoxy)-5-oxopentanoic acid (( t

[0136] Bu)3NODAGA-COOH, 100 mg, 0.19 mmol, 1.0 equiv), N,N,N',N'-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU, 122 mg, 0.3 mmol, 1.2 equiv) and N-hydroxysuccinimide (NHS, 37.4 mg, 0.3 mmol, 1.2 equiv) were dissolved in acetonitrile (10 mL) and stirred at room temperature for 24 hours. After removing the solvent under reduced pressure, the resulting residue was redissolved in dichloromethane (10 mL) and then quickly washed with water (3×4 mL). The organic layer was dried over magnesium sulfate, filtered and evaporated to give the title product as a white foam (141 mg, 0.22 mmol, 85%).

[0583] ( t Synthesis of Bu3NODAGA peptide: Will( t Bu)3NODAGA-NHS (1.5 eq.) was added to a solution of each peptide (1 eq.) in dimethylformamide (1 mL). The pH of each solution was adjusted to 6.5 using diisopropylethylamine (DIPEA), and the mixture was stirred at room temperature for 24 h. The reaction was monitored by LCMS. After completion of the reaction, ( t The cyclic peptide was purified by reverse phase preparative HPLC on a C-5 column (Luna, 10μ, 250×21.2 mm) using a gradient of 5% mobile phase A (0.1% TFA in water) to 60% mobile phase B (0.1% TFA in acetonitrile) in 45 minutes. The purified compound was lyophilized to give the title product.

[0584] Synthesis of Compound 16, Compound 17, Compound 18, Compound 19, Compound 20 and Compound 21: In a reaction vessel, about 10 mg of ( t The NODAGA peptide was deprotected in a solution of 1 mL TFA, methanesulfonic acid, 1-dodecanethiol and H2O (92:3:3:2). Various reaction mixtures were stirred for 2 hours and analyzed by LCMS. After the reaction was complete, cold ether (15 mL) was added to precipitate a solid. The mixture was centrifuged to remove the supernatant. The solid was washed with ether and dried to obtain the product as a white solid.

[0585] Compound 16: C 77 H 107 C1N 16 O 25 S2, MS (ESI) calculated: 878.3 [(M+2H) / 2] 2+ ; Measured: 878.4.

[0586] Compound 17: C 77 H 107C1N 16 O 25 S2, MS (ESI) calculated: 878.3 [(M+2H) / 2] 2+ ; Measured: 878.5.

[0587] Compound 18: C 74 H 105 C1N 18 O 24 S2, MS (ESI) calculated: 866.2 [(M+2H) / 2] 2+ ; Actual measurement: 866.1.

[0588] Compound 19: C 74 H 105 C1N 18 O 24 S2, MS (ESI) calculated: 866.1 [(M+2H) / 2] 2+ ; Measured: 866.3.

[0589] Compound 20: C 74 H 105 C1N 18 O 24 S2, MS (ESI) calculated: 866.1 [(M+2H) / 2] 2+ ; Actual measurement: 866.1.

[0590] Compound 21: C 75 H 108 C1N 17 O 23 S2, MS (ESI) calculated: 857.9 [(M+2H) / 2] 2+ ; Measured: 858.7

[0591] Compound 22: C 74 H 105 C1N 18 O 24 S2, MS (ESI) calculated: 866.3 [(M+2H) / 2] 2+ ; Actual measurement: 866.1.

[0592] Table 3. NODAGA modified peptides

[0593]

[0594] Compound 68 Ga-16, 68 Ga-17, 68 Ga-18, 68 Ga-19, 68 Ga-20 and 68 Ga-21 and 68 Radiochemistry of Ga-22 synthesis: 68 GaCl3 (1 mCi in 0.5 mL HCl (0.6 M) solution) was diluted with 3 M sodium acetate (200 μL) at pH 5 to reach pH 4.1. 68The GaCl3 solution (200 μL) was mixed with a solution of various NODAGA peptides (0.1 mM, 10 μL) in sodium acetate (10 mM, pH 4.1), and the reaction mixture was heated at 60°C for 10 min and purified by Sep-Pak Light C 18 A column (Waters) was used to remove any radioactive metal impurities (germanium-68 breakthrough). The compound was determined by radio-HPLC analysis. 68 Ga-16, compound 68 Ga-17, compound 68 Ga-18, compound 68 Ga-19, compound 68 Ga-20( Figure 8 ), compound 68 Ga-21 and compounds 68 The radiochemical purity of the final solution of Ga-22 was ≥ 95%.

[0595] Compound 64 Radiochemical synthesis of Cu-20: 64 A solution of CuCl2 (1 mCi) in 0.5 mL HCl (0.6 M) was diluted with 3 M sodium acetate (200 μL) to pH 4.5. A solution of NODAGA peptide 22 (0.1 mM, 10 μL) in sodium acetate (10 mM, pH 4.5) was added and heated at 60°C for 10 minutes. The compound was confirmed by radio-HPLC analysis. 64 The radiochemical purity of Cu-20 is ≥95%.

[0596] DOTAGA Chelating Agents

[0597] ( t Synthesis of Bu4DOTAGA-PFP: Will( t Bu)4DOTAGA (200 mg, 0.29 mmol) and pentafluorophenol (88 mg, 0.48 mmol) were dissolved in dichloromethane (1 mL), and PS-DCC (286 mg, 0.48 mmol, 1.67 mmol / g) was added to the mixture. The reaction was stirred at room temperature and monitored by HPLC. After the reaction was completed, the resin was removed by filtration. The filtrate was evaporated and the residue was dried under reduced pressure. The crude product ( t Bu)4DOTAGA-PFP was used in the next step without further purification. ([M+H] + Theoretical MW = 868.0, observed 867.7).

[0598] ( t Synthesis of Bu4DOTAGA peptide: ( tBu)4DOTAGA-PFP (50 mg, 0.06 mmol) and peptide (79.2 mg, 0.06 mmol) were dissolved in DMF (1 mL) and the pH was kept at about 6.5 by adding DIPEA. The reaction was monitored by HPLC. After the reaction was completed, the solid was precipitated with saline, washed with distilled water and dried under reduced pressure. The crude product was used in the next step without further purification. ([M+H] + The theoretical MW = 2056.9, the observed MW is 2056.7).

[0599] Synthesis of compound 23: The reaction mixture was stirred in a mixture of TFA, triisopropylsilane, dodecanethiol, MSA and water (92:2:2:2:2, 1 mL) t The tert-butyl ester was cleaved by adding 4-(4-(4-Bu)-4-DOTAGA peptide and overnight at room temperature. The reaction was monitored by HPLC. The reaction mixture was precipitated in cold ether. The solid was filtered, washed with cold ether and dried under reduced pressure. The crude product was dissolved in water (6 mL) and the cyclic peptide was purified in 30 minutes by reverse phase preparative HPLC on a C-18 column (Luna, 10μ, 250×21.2mm) using a gradient of 5% mobile phase A (0.1% TFA aqueous solution) to 95% mobile phase B (0.1% TFA acetonitrile solution). The fractions of pure compound 23 were combined and lyophilized to obtain a white powder.

[0600] Compound 23:C 78 H 112 C1N 19 O 26 S2, MS (ESI) calculated: 1832.43 [M+H] + ,916.7[(M+2H) / 2] 2+ ; Measured: 1831.7[M+H] + ,916.2[(M+2H) / 2] 2+ .

[0601] Table 5. DOTAGA modified peptides

[0602]

[0603] Compound 68 Radiochemical synthesis of Ga-23: SCX cartridge (100 mg, particle size 40 μm (Agilent, catalog number 12102013)) was pre-treated by washing with 5.5 M HCl (1 mL) and then with water (10 mL). Ga-68 (3.0 mCi) was eluted with 4 mL of 0.05 M HCl and loaded onto the pre-treated SCX cartridge. The cartridge was purged with air and eluted with 0.3 mL of 3 M NaCl solution (containing 0.1 M HCl). 68GaCl3. 0.15mL 68 GaCl3 was added to a solution of compound 23 (25 μL, 1.0 mM) mixed with 0.15 mL 0.15 M NaOAc buffer. The reaction mixture was heated to 90°C for 10 minutes and analyzed by radio-HPLC. Radiochemical purity was ≥99% as determined by radio-HPLC analysis.

[0604] Compound 90 Radiochemical synthesis of Y-23: Containing 0.5 mCi 90 100 μL of YCl3 solution was added to a solution of compound 23 (25 μL, 1.0 mM) and mixed with 0.15 mL of sodium acetate buffer at pH 5. The reaction mixture was heated to 40°C for 60 minutes and analyzed by radio-HPLC. HPLC fractions were gamma counted.

[0605] Compound 177 Radiochemical synthesis of Lu-23: Containing 0.5 mCi 177 100 μL of LuCl3 solution was added to a solution of compound 23 (25 μL, 1.0 mM) and mixed with 0.15 mL of sodium acetate buffer at pH 5. The reaction mixture was heated to 40°C for 60 minutes and analyzed by radio-HPLC.

[0606] Compound 225 Radiochemical synthesis of Ac-23: The 0.1 mCi 225 100 μL of a solution of Ac(NO3)3 and 20% L-ascorbic acid was added to a solution of compound 23 (25 μL, 1.0 mM) and mixed with 0.15 mL of Tris buffer at pH 6. The reaction mixture was heated to 60°C for 60 minutes and analyzed by radio-HPLC, and the HPLC fractions were gamma counted.

[0607] NOTA Chelating Agents

[0608] ( t Synthesis of Bu)2NOTA peptide: NOTA tBu)2 (50 mg, 0.12 mmol) and HATU (68.6 mg, 0.18 mmol) were dissolved in DMF (1 mL), and the mixture was stirred at room temperature for 30 minutes. The peptide (181 mg, 0.13 mmol) was dissolved in DMF (1 mL), and DIEA was added to maintain the pH at 6.5. The reaction was monitored by LC-MS. After the reaction was completed, the mixture was diluted with water (4 mL) and the cyclic peptide was purified in 30 minutes by reverse phase preparative HPLC on a C-18 column (Luna, 10 μ, 250×21.2 mm) using a gradient of 5% mobile phase A (0.1% TFA aqueous solution) to 95% mobile phase B (0.1% TFA acetonitrile solution). The fractions of the pure compound were combined and lyophilized to obtain the final product. ([M+H] + The theoretical MW = 1771.5, the observed MW is 1771.7).

[0609] Synthesis of compound 24: In a reaction vessel, about 35 mg of ( t Bu)2NOTA peptide was deprotected in a solution of 1mL TFA, methanesulfonic acid, 1-dodecanethiol, triisopropylsilane and H2O (92:2:2:2:2). The reaction mixture was stirred at room temperature for 2 hours and analyzed by LCMS. After the reaction was complete, cold ether (15mL) was added to precipitate a solid. The mixture was centrifuged and the supernatant was removed. The solid was washed with ether and dried to give the product as a white solid. The crude product was dissolved in water (5mL) and the cyclic peptide was purified in 30 minutes by reverse phase preparative HPLC on a C-18 column (Luna, 10μ, 250×21.2mm) using a gradient of 5% mobile phase A (0.1% TFA aqueous solution) to 95% mobile phase B (0.1% TFA acetonitrile solution). The fractions of pure compound 24 were combined and lyophilized to obtain a white powder.

[0610] Compound 24: C 71 H 101 C1N 18 O 22 S2, MS (ESI) calculated: 1659.3 [M+H] + ,830.1[(M+2H) / 2] 2+ ; Measured: 1659.7[M+H] + ,829.7[(M+2H) / 2] 2+ .

[0611] Table 4. NOTA modified peptides

[0612]

[0613] Compound 68 Radiochemical synthesis of Ga-24:SCX cartridge (100 mg, particle size 40 μm (Agilent, catalog number 12102013)) was pre-treated by washing with 5.5 M HCl (1 mL) and then with water (10 mL). Ga-68 (3.2 mCi) was eluted with 4 mL of 0.05 M HCl and loaded onto the pre-treated SCX cartridge. The cartridge was purged with air and eluted with 0.3 mL of 3 M NaCl solution (containing 0.1 M HCl). 68 GaCl3. 68 GaCl3 (0.15 mL) was added to a solution of compound 24 (25 mL, 0.6 mM) in NaOAc buffer (1.5 M, 0.15 mL, pH 4.5). The reaction mixture was heated to 90°C for 10 minutes and analyzed by radio-HPLC. The radiochemical purity of the final product was ≥98%.

[0614] Compound A1 18 Radiochemical synthesis of F-24: A Sep-Pak Light Accell Plus QMA cartridge (Waters) was preconditioned by passing 10 mL of 0.4 M KHCO3 followed by 10 mL of DI water. 18 F (2.0 mCi, 200 μL) was loaded onto the column and washed with DI water (5 mL), after which it was eluted from the column with KHCO3 (400 μL) and acidified to pH 4.0 with acetic acid. A stock solution of AlCl3 (2 mM, pH 4, 0.1 M sodium acetate buffer) was prepared and 300 μL was taken and mixed with 18 Compound 24 (2 mM, 300 μL) in 0.1 M NaOAc was added and heated at 115°C for 15 min. The mixture was analyzed by radio HPLC (Ultra AQ, C 18 , 5μm, 250×4.6mm), using a gradient of 5% mobile phase A (50mM ammonium acetate in water) to 95% mobile phase B (10% 50mM ammonium acetate in water and 90% acetonitrile). The radiochemical purity of the product was 80%. After HPLC purification, the radiochemical purity was >95%.

[0615] Example 8: Evaluation of fibrin-specific compounds in a rat carotid endothelial injury model

[0616] In vivo studies were performed in a rat carotid endothelial injury model, in which the common carotid artery was isolated and briefly crushed with a hemostatic agent, resulting in the formation of a mural thrombus at the site of crushing. Afterwards, vessel wall fibrin was imaged with our PET compounds. All six compounds were evaluated in this model.

[0617] Adult male Wistar rats were anesthetized with isoflurane (1-2% in medical air). The body temperature was maintained at 37°C using a thermoregulated heating pad. Compound injection and blood sampling were performed by cannulation of the right femoral vein and artery, respectively. Endothelial injury was induced by causing a crush injury to the right common carotid artery (1-2 cm proximal to the carotid bifurcation) by clamping the vessel with a hemostat for 5 minutes. Rats were placed in a micro PET / CT scanner immediately after surgery. Compounds (200-600 μL) were injected, PET-CT images were collected, and blood samples were drawn from the femoral artery before and 2, 5, 10, 15, 30, 60, 90, and 120 minutes after compound injection. Tissues were finally collected for in vitro analysis.

[0618] Example 9: Blood Analysis to Determine Blood Clearance of Compounds in Rats

[0619] Blood samples were drawn before and 2, 5, 10, 15, 30, 60, 90, and 120 minutes after probe injection. Each blood sample was weighed and counted using a gamma counter. The activity in each sample was calculated as the percentage of injected dose per gram of tissue (%ID). 68 Ga-19 and 68 Ga-20 exhibits faster blood clearance and better metabolic stability ( Fig. 9 ).

[0620] Example 10: Functional Compound Testing Assay for Assessing the Fraction of Intact Compound Circulating at Various Time Points

[0621] Plasma was separated (at t = 10 and 60 min) and the plasma samples were incubated with fibrin immobilized in the wells for 2 h at room temperature. After incubation, counts in the supernatant from the fibrin-containing wells and the empty wells were measured on a gamma counter and divided by the plasma weight to determine the concentrations of unbound probe [unbound] and total probe [total], respectively. 68 The amount of Ga [bound] was calculated as [bound] = [total] - [unbound]. As a positive control, an aliquot of the dose was spiked into plasma and used to estimate the total possible fibrin binding in the assay (% bound at t = 0). The amount of functional probe in the blood at time t was calculated by taking the ratio of the % fibrin binding at time t to the % bound at t = 0 and multiplying this ratio by the total amount measured in the blood. 68 The percentage of activity bound to fibrin was estimated by measuring the activity in the wells containing fibrin and comparing it to the activity in the wells without fibrin. This % binding was compared to the % binding measured when the pure compound was spiked into fresh plasma ( Fig.10 and Fig.11 ).

[0622] Example 11: Radio-HPLC analysis of blood for identification of metabolites

[0623] Aliquots of the 15 and 90 minute plasma samples were injected into the HPLC and fractions were collected every 30 seconds. The counted fractions were compared with pure compound injected into the same column to determine the number of metabolites and the fraction of intact compound circulating at each time point. Unexpectedly, compound 68 Ga-16 rapidly forms metabolites after injection, while other compounds of similar structure remain intact ( Fig.12 ).

[0624] Example 12: Biodistribution

[0625] The injured carotid artery, contralateral carotid artery and all organs were removed, weighed and counted using a gamma counter. The activity in various organs is shown as the percentage of injected dose (%ID) per gram of tissue ( Fig.13 ).

[0626] Example 13: Autoradiography

[0627] The injured ipsilateral and contralateral carotid arteries were mounted on multifunctional film and imaged using the Perkin-Elmer CyclonePlus Storage Phosphor System. 68 Ga-19 (n=6 rats) and 68 Additional in vivo studies were performed with Ga-20 (n = 9 rats) Fig.14 ). Among them, the compound 68 Ga-20 exhibited faster blood clearance, better metabolic stability, and a better ipsi:opposite side ratio.

[0628] Compound 68 Ga-20 was also successfully validated in a rabbit high-risk plaque model (plaque rupture model). After in vivo PET, the mean SUV values, TBRvc, and TBRm of all rabbits were compared. Fig.16 Mean group PET uptake (SUV) and ratios at different time points are shown. As expected, the ratio of plaque rupture to control SUV was higher with increasing imaging time points after injection. Fig.17 Representative PET-MR images from plaque ruptured and control animals are shown. Fibrin clots are seen as points of uptake along the aorta in the plaque ruptured group, while the control aorta shows a very uniform outline.

[0629] Example 14: In vitro study of human carotid endarterectomy specimens

[0630] Twelve discarded surgical specimens were obtained from asymptomatic patients undergoing elective carotid endarterectomy at Massachusetts General Hospital and used for the in vitro studies described herein. The samples were processed as described above (Examples 6 and 13) for histology, autoradiography, and probe binding assays.

[0631] Discarded surgical specimens were embedded in optimal cutting temperature compound, snap frozen, and stored at -80°C prior to further analysis. Alternate serial cryosectioning was performed for histology and autoradiography; three tissue samples separated by 50 μm were used for these experiments. The remaining tissue samples were processed for functional probe assays.

[0632] For histology: Specimens were fixed using 4% paraformaldehyde containing 30% sucrose. The rostral portion of each segment was embedded in optimal cutting temperature compound for histological frozen sections (Carstairs stain). The remaining aortic segments were dissected for gross pathological examination. Carstairs stain was used to distinguish fibrin (bright red), collagen (bright blue), erythrocytes (yellow to orange), platelets (gray blue to navy blue), and muscle (red). Sections were examined using a microscope (NikonEclipse 50i, Kodak Scientific Imaging System), and digital images were acquired using a camera connected to a computer (SPOT 7.4Slider RTKE, Diagnostic Instruments).

[0633] For autoradiography, three 30 μm thick sections were 68 Ga-20 or non-fibrin binding control probe 68 Ga-22 was incubated for 45 min at room temperature on a laboratory shaker. Autoradiography was performed after three washes in PBS. The exposure time was 2 min.

[0634] For functional probe assays, samples were thawed at room temperature, weighed (8-15 mg per sample), cut into duplicates, and placed in 1 mL PBS containing 111-148 kBq (3-4 μCi) 68 Ga-20 or non-fibrin binding control probe 68 Ga-22 in a test tube. The mixture was shaken at room temperature for 45 minutes. After centrifugation, the supernatant (sup-1) was removed and retained for further analysis, and the tissue samples were washed with PBS (1 mL); this procedure was repeated twice for each sample. The wash solutions (wash-1 and wash-2) were retained for further analysis. The activity in the tissue samples and solutions sol-1, wash-1, and wash-2 was measured on a gamma counter. The percentage uptake was calculated as follows:

[0635] Uptake % = [(activity in tissue) / total activity (tissue + sup-1 + wash-1 + wash-2)] × 100

[0636] result:

[0637] Autoradiography ( Fig.18 A-18B, 18G-18H and 18M) and functional probe assays ( Fig.18 N) showed no binding to the non-binding probe 68 Significantly higher than Ga-22 68 Ga-20 uptake (P < 0.05 for both). 68 There was significant interpatient variability in Ga-20 uptake ( Fig.18 M-18N). Carstairs staining results were consistent with autoradiography and probe testing. In some patient specimens, high 68 Ga-20 uptake and the presence of large amounts of fibrin ( Fig.18 A-18F), while low uptake samples had relatively less fibrin ( Fig.18 G-18L). On the other hand, the results obtained from most specimens were not so clear-cut, and the total amount of fibrin in the tissue did not correlate with either the autoradiography (P = 0.45) or the binding test (P = 0.28).

[0638] Other Implementations

[0639] It should be understood that although the invention has been described in conjunction with specific embodiments, the foregoing description is intended to illustrate rather than limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages and improvements are within the scope of the claims.

Claims

1. Compounds of formula IV: or a pharmaceutically acceptable salt thereof, wherein R 4 It is a radioactive isotope; C 4 is a chelating moiety selected from the group consisting of: AA-CP 4 For –y–e–C–Hyp–Y(3–Cl)–G–L–C–H–I–Q– –y–e–C–Hyp–Y(3–Cl)–G–L–C–H–I–l– –Y–e–C–Hyp–Y(3–Cl)–G–L–C–H–I–Q– –Y–e–C–Hyp–Y(3–Cl)–G–L–C–H–I–q–or –y–e–C–Hyp–Y(3–Cl)–G–L–C–Y–I–Q–; L 4 For the joint; y is 1; and z is 0.

2. The compound according to claim 1, wherein R 4 The radioisotope is selected from therapeutic radioisotopes and radioisotopes detectable using nuclear imaging techniques.

3. The compound according to claim 2, wherein Radioisotopes that can be detected using nuclear imaging techniques are positron emitting isotopes or radioisotopes suitable for single photon emission computed tomography (SPECT) imaging.

4. The compound according to claim 3, wherein The positron emitting isotope is selected from fluorine-18, aluminum fluoride (Al 18 F), scandium-43, scandium-44, manganese-51, manganese-52, copper-60, copper-61, copper-62, copper-64, gallium-68, yttrium-86, zirconium-89, iodine-124, terbium-149 and terbium-152.

5. The compound according to claim 3, wherein Radioisotopes suitable for SPECT imaging are selected from the group consisting of gallium-67, technetium-99m, indium-111, iodine-123, iodine-125, terbium-155, and lead-203.

6. The compound according to claim 2, wherein The therapeutic radioisotope is selected from scandium-47, copper-67, yttrium-90, iodine-131, samarium-153, terbium-161, holmium-166, lutetium-177, rhenium-188, astatine-211, lead-212, bismuth-213, radium-223, actinium-225 and thorium-227.

7. The compound according to claim 1, wherein AA-CP 4 yes 8. The compound according to claim 1, wherein AA-CP 4 yes 9. The compound according to claim 1, wherein C 4 Select from the following group:

10. The compound according to claim 9, wherein C 4 for 11. The compound according to claim 1, wherein C 4 Select from the following group:

12. The compound according to claim 1, wherein C 4 Select from the following group:

13. The compound according to claim 1, wherein L 4 It is pyridyl or (pyridyl)-C(O)-.

14. The compound according to claim 1, wherein R 4 Selected from aluminum fluoride (Al 18 F), scandium-43, scandium-44, scandium-47, manganese-51, manganese-52, copper-60, copper-61, copper-62, copper-64, copper-67, gallium-67, gallium-68, yttrium-86, zirconium-89, technetium-99m, yttrium-90, indium-111, terbium-149, terbium-152, samarium-153, terbium-155, terbium-161, holmium-166, lutetium-177, rhenium-188, lead-203, lead-212, bismuth-213, radium-223, actinium-225, and thorium-227.

15. The compound according to claim 1, wherein R 4 Selected from fluorine-18, iodine-123, iodine-124, iodine-125, iodine-131 and astatine-211.

16. The compound according to claim 1, wherein The compound is selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

17. The compound according to claim 1, wherein The compound is selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

18. The compound according to claim 1, wherein The compounds are: or a pharmaceutically acceptable salt thereof.

19. A pharmaceutical composition comprising the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

20. The pharmaceutical composition of claim 19, comprising a free radical scavenger.

21. The pharmaceutical composition according to claim 20, wherein Free radical scavengers are antioxidants.

22. The pharmaceutical composition of claim 20, wherein The free radical scavenger is selected from carnosic acid, green tea extract, apigenin, diosmin, rosmarinic acid, lipoic acid, beta-carotene, L-ascorbic acid (vitamin C), N-acetylcysteine ​​(NAC), delta-tocopherol, rutin, amifostine, resveratrol, gentisic acid and gallic acid.

23. Use of the pharmaceutical composition according to any one of claims 19 to 22 or the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for imaging fibrin in a mammal, the use comprising: a) administering to a mammal an effective amount of the pharmaceutical composition according to any one of claims 19 to 22 or an effective amount of the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof; b) using nuclear imaging techniques to obtain images of mammalian fibrin; c) obtaining anatomical images of mammals using magnetic resonance imaging or computed tomography; and d) superimposing the images of steps b) and c) to locate the fibrin image within the anatomical image of the mammal.

24. The use according to claim 23, wherein: The presence of fibrin is associated with neuroinflammation.

25. The use according to claim 23, wherein: Neuroinflammation is associated with Alzheimer's disease, multiple sclerosis or traumatic brain injury.

26. The use according to claim 23, further comprising: e) administering an effective amount of the pharmaceutical composition of any one of claims 19 to 22 or an effective amount of the compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 4 It is a therapeutic radioisotope.

27. The use according to claim 23, wherein: Fibrin is present in tumors.

28. The use according to claim 27, wherein: Tumors are cancerous.

29. The use according to claim 23, wherein: Fibrin is present in blood clots.

30. Use of the pharmaceutical composition according to any one of claims 19 to 22 or the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a disease or condition associated with fibrin present in a mammal, the use comprising: Administering to a mammal an effective amount of the pharmaceutical composition of any one of claims 19 to 22 or an effective amount of the compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 4 It is a therapeutic radioisotope.

31. Use of the pharmaceutical composition according to any one of claims 19 to 22 or the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer in a mammal, the use comprising: Administering to a mammal an effective amount of the pharmaceutical composition of any one of claims 19 to 22 or an effective amount of the compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 4 It is a therapeutic radioisotope.

32. The use according to claim 31, wherein: R as a therapeutic radioisotope 4 Selected from scandium-47, copper-67, yttrium-90, iodine-131, samarium-153, terbium-161, holmium-166, lutetium-177, rhenium-188, astatine-211, lead-212, bismuth-213, radium-223, actinium-225 and thorium-227.

33. Use of the pharmaceutical composition according to any one of claims 19 to 22 or the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for detecting and treating a disease or condition associated with fibrin present in a mammal, the use comprising: a) administering to a mammal an effective amount of the pharmaceutical composition according to any one of claims 19 to 22 or an effective amount of the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 4 are radioactive isotopes that can be detected using nuclear imaging techniques; b) using nuclear imaging techniques to obtain images of mammalian fibrin; c) obtaining anatomical images of mammals using magnetic resonance imaging or computed tomography; d) superimposing the images of steps b) and c) to locate the fibrin image within the anatomical image of the mammal; and e) administering an effective amount of the pharmaceutical composition of any one of claims 19 to 22 or an effective amount of the compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 4 It is a therapeutic radioisotope.

34. Use of the pharmaceutical composition according to any one of claims 19 to 22 or the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for detecting and treating a disease or condition associated with fibrin present in a mammal, the use comprising: a) administering to a mammal an effective amount of the pharmaceutical composition according to any one of claims 19 to 22 or an effective amount of the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 4 is a radioactive isotope detectable using nuclear imaging techniques, selected from the group consisting of fluorine-18, copper-64, and gallium-68; b) using nuclear imaging techniques to obtain images of mammalian fibrin; c) obtaining anatomical images of mammals using magnetic resonance imaging or computed tomography; d) superimposing the images of steps b) and c) to locate the fibrin image within the anatomical image of the mammal; e) administering an effective amount of the pharmaceutical composition of any one of claims 19 to 22 or an effective amount of the compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 4 is a therapeutic radioisotope selected from the group consisting of yttrium-90, lutetium-177 and actinium-225.

35. The use according to claim 34, wherein: Fibrin is present in tumors.

36. The use according to claim 35, wherein: Tumors are cancerous.

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