Diagnostic methods for amyloid disease

Abstract

This disclosure relates, in some embodiments, to methods and compositions for detecting amyloid-related diseases in individuals.

Description

[Technical Field]

[0001] Cross-references to related applications This application claims priority under U.S. Provisional Patent Application No. 63 / 581,213 filed on 7 September 2023 and U.S. Provisional Patent Application No. 63 / 657,715 filed on 7 June 2024 (both of which are incorporated herein by reference).

[0002] Reference to electronic sequence lists The entire contents of the electronic sequence listing (165992001240SEQLIST.xml; size: 40,979 bytes; and creation date: September 6, 2024) are incorporated herein by reference.

[0003] field This disclosure relates, in some aspects, to a method for diagnosing amyloid-related diseases. [Background technology]

[0004] background Systemic amyloidosis is a progressive protein misfolding disorder characterized by the extracellular deposition of protein fibrils, extracellular matrix components, and serum proteins. Amyloid accumulation leads to organ failure, reduced quality of life, and ultimately death, particularly in the heart and kidneys (the organs most affected in all types of amyloidosis). While approximately 20 different proteins have been identified as fibril components in systemic amyloidosis (Aimo A, Merlo M, Porcari A et al., Eur J Heart Fail. 2022), patients with amyloidosis associated with mutant or wild-type transthyretin (ATTRv and ATTRwt), immunoglobulin light chain (AL), and leukocyte chemotactic factor 2 (ALECT2) account for over 90% of cases diagnosed in the United States (Edwards CV, Rao N, Bhutani D et al., Blood. 2021;138:2632-2641).

[0005] Cardiac amyloidosis is an unsettling sign in both AL-related and ATTR-related amyloidosis, causing restrictive hypertrophic cardiomyopathy, particularly of the left ventricular wall and interventricular septum, accompanied by abnormalities in electrical conduction and strain (Garcia-Pavia P, Aus Dem Siepen F, Donal E et al., N Engl J Med. 2023:NEJMoa2303765). However, any organ or tissue can be affected, leading to a heterogeneous clinical presentation and making accurate and rapid diagnosis difficult (Dorbala S, Cuddy S, Falk RH. JACC Cardiovasc Imaging. 2020;13:1368-1383).

[0006] Non-invasive imaging is particularly useful in diagnostic algorithms for cardiac amyloidosis (Dorbala S, Park MA, Cuddy S et al., J Nucl Med. 2021;62:716-722). While echocardiography and cardiac magnetic resonance imaging (CMR) can reveal structural abnormalities and functional sequelae associated with amyloidosis, these techniques are not amyloid-specific. 99m Tc-Pyrophosphate ( 99m TcPyP) or 99m Nuclear imaging using Tc-3,3-diphosphono-1,2-propanedicarboxylic acid (DPD) is routinely used in the diagnosis of cardiac ATTR amyloidosis. These osteoaffinic substances bind to cardiac microcalcifications associated with cardiac amyloid, preferentially detecting only ATTR-associated cardiac amyloidosis (Antoni G, Lubberink M, Estrada S et al., J Nucl Med. 2013;54:213-220). In some embodiments, existing methods are limited by the lack of amyloid-specific indicators for monitoring changes in amyloid load over time. Therefore, non-invasive and quantitative indicators for detecting and monitoring cardiac and extracardiac amyloid deposition would be clinically beneficial. This specification provides methods and compositions to address such and other requirements. [Overview of the project]

[0007] General In one embodiment, the Specified provides a method for diagnosing an early stage of amyloid-related disease in an individual at risk of developing amyloid-related disease, the method comprising: a) administering to the individual an amyloid-reactive peptide comprising an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label; and b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the tissue or organ of the individual, wherein an amount of the amyloid-reactive peptide exceeding a threshold indicates that the individual has amyloid-related disease, and the amyloid-related disease is an early stage of amyloid-related disease.

[0008] In some embodiments, individuals are determined to be at risk of amyloid-related disease based on the presence of a gene mutation, having multiple myeloma, having amyloid-positive laminectomy tissue, having amyloid-positive tissue derived from carpal tunnel release, having monoclonal immunoglobulinemia of unknown significance (MGUS), having heart failure with preserved ejection fraction (HFpEF), having heart failure with reduced ejection fraction (HFrEF), being from a susceptible ethnic group, or being elderly. In some embodiments, the gene mutation is located in the transthyretin protein. In some embodiments, the gene mutation is located in the fibrinogen-alpha protein.

[0009] In some embodiments, the individual does not have symptoms of amyloidosis. In some embodiments, the individual has neurological symptoms of amyloid disease. In some embodiments, the individual has been previously diagnosed as not having amyloid-related disease.

[0010] In another embodiment, this specification provides a method for diagnosing an early stage of amyloid-related disease in an individual suspected of having amyloid-related disease, the method comprising: a) administering to the individual an amyloid-reactive peptide comprising an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label; and b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the tissue or organ of the individual, wherein an amount of the amyloid-reactive peptide exceeding a threshold indicates that the individual has amyloid-related disease, and the amyloid-related disease is an early stage of amyloid-related disease.

[0011] In another embodiment, this specification provides a method for determining the prognosis of an individual having an early-stage amyloid-related disease, the method comprising: a) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs to quantify the amyloid in the individual at a first time point; b) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label, and detecting the amount of the detectable label in the individual's tissues or organs to quantify the amyloid in the individual at a second time point; and c) determining the prognosis of the individual by comparing the amyloid quantified at the first time point in the tissues or organs with the amyloid quantified at the second time point in the tissues or organs, wherein the amyloid-related disease is an early-stage amyloid-related disease.

[0012] In some embodiments, the prognosis of an individual with amyloid-related disease is based on the detection of amyloid-reactive peptide levels in the heart only. In some embodiments, the prognosis of an individual with amyloid-related disease is based on the detection of amyloid-reactive peptide levels in the heart and kidneys. In some embodiments, the prognosis of an individual with amyloid-related disease is based on the detection of amyloid-reactive peptide levels in the heart, kidneys and all other organs.

[0013] In another embodiment, this specification provides a method for treating amyloid-related disease in an individual having an early-stage amyloid-related disease, the method comprising: a) administering to the individual an amyloid-reactive peptide comprising an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label; b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs; and c) if the amount of the amyloid-reactive peptide exceeds a threshold, performing treatment for the amyloid-related disease.

[0014] In some embodiments, the individual is suspected to have an amyloid-related disease.

[0015] In another embodiment, the Specified provides a method for monitoring the treatment of amyloid-related disease in an individual having an early-stage amyloid-related disease, the method comprising: a) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label to the individual and quantifying the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs; b) administering treatment for the amyloid-related disease; c) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label to the individual and quantifying the amount of the amyloid in the individual's tissues or organs by detecting the amount of the detectable label; and d) determining whether the treatment is effective by comparing the amyloid quantified in the tissues or organs at the first time point with the amyloid quantified in the tissues or organs at the second time point.

[0016] In some embodiments, the method further comprises administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label to an individual, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs to quantify amyloid in the individual at a second, optionally third, fourth, and / or fifth time point.

[0017] In some embodiments, the first time point and the second time point are separated by at least six weeks.

[0018] In another embodiment, this specification provides a method for selecting a treatment for amyloid-related disease in an individual, the method comprising: a) administering to the individual an amyloid-reactive peptide comprising an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label; and b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the tissue or organ of the individual, wherein if amyloid is detected in the heart, a treatment for amyloid-related disease is performed, and if amyloid is not detected in the heart, an alternative therapy is performed.

[0019] In another embodiment, the Specified provides a method for managing the treatment of an amyloid-related disease in an individual, the method comprising: a) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label, and quantifying the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs; b) administering treatment for the amyloid-related disease; c) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label, and quantifying the amount of the amyloid in the individual's tissues or organs by detecting the amount of the detectable label in the individual's tissues or organs; d) comparing the amyloid quantified in the tissues or organs at the first time point with the amyloid quantified in the tissues or organs at the second time point; and e) coordinating the treatment for the amyloid-related disease.

[0020] In some embodiments, if the amyloid disease is stable between the first and second time points, the treatment is modified. In some embodiments, if the amount of amyloid increases, the treatment is restarted.

[0021] In some embodiments, the amount of amyloid-reactive peptide that exceeds a threshold indicates that the individual has an amyloid-related disease. In some embodiments, the amyloid-related disease is an early stage of an amyloid-related disease.

[0022] In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 13.

[0023] In some embodiments, the detectable label is a radioactive label. In some embodiments, the detectable label is 11 C, 18 F, 111 In, 99m Tc, 89 Zr, 68 Ga and 123 I, 124 I and 125 I. In some embodiments, the detectable label is selected from the group consisting of 124 I.

[0024] In some embodiments, the method further comprises administering to the individual the amyloid-reactive peptide labeled with I at a dose of about 0.1 to about 25 mCi of 124 . In some embodiments, the method further comprises administering to the individual the amyloid-reactive peptide labeled with I at a dose of about 0.3 mCi, 1 mCi or about 2 mCi of 124 . In some embodiments, the method further comprises administering to the individual the amyloid-reactive peptide labeled with I at a dose of about 1 mCi (±10%) of 124 .

[0025] In some embodiments, the method further comprises administering Tc-99 to the individual at a dose of about 20 mCi.

[0026] In some embodiments, the method further comprises administering about 0.3 mg, 1 mg, 1.5 mg or about 2 mg of the amyloid-reactive peptide to the individual.

[0027] In some embodiments, the amyloid-reactive peptide is administered intravenously.

[0028] In some embodiments, the method further includes determining organ-specific standard intake ratios for organs within an individual. In some embodiments, the organ-specific SUVR of an individual is selected from the group consisting of mean SUVR, maximum SUVR, and peak SUVR.

[0029] In some embodiments, the method further includes determining the organ-specific SUV of an individual. In some embodiments, the organ-specific SUV of an individual is selected from the group consisting of the mean SUV, the maximum SUV, and the peak SUV.

[0030] In some embodiments, the method further includes determining the organ-specific percentage injection volume per gram (%ID) of the individual.

[0031] In some embodiments, the sensitivity when detecting amyloid using amyloid-reactive peptides is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%. In some embodiments, the sensitivity is about 99% or about 100%.

[0032] In some embodiments, the specificity of the amyloid-reactive peptide is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%. In some embodiments, the specificity of the amyloid-reactive peptide is approximately 99% or 100%.

[0033] In some embodiments, the method further includes determining one or more health-related quality of life indicators for an individual.

[0034] In some embodiments, the method further includes detecting one or more biomarkers associated with amyloid-related disease. In some embodiments, the biomarkers associated with amyloid-related disease are selected from the group consisting of troponin T, NTproBNP, urinary protein levels, UACR, EGFR, and alkaline phosphatase levels.

[0035] In some embodiments, the tissue or organ of an individual is selected from the group consisting of lungs, fat, heart, kidneys, pancreas, joints, spine, liver, spleen, adrenal gland, bone lesions, choroid plexus, pituitary gland, uterus, bone marrow, musculoskeletal tissue, gastrointestinal tract, and prostate.

[0036] In some embodiments, the threshold is the mean SUVR threshold. In some embodiments, when the organ or tissue is the liver, the mean SUVR threshold is approximately 1.0, 1.1, 1.2, 1.3, 1.4, 1.5 or 1.6; when the organ or tissue is the spleen, the mean SUVR threshold is approximately 0.9, 1.0, 1.1, 1.2, 1.3, 1.4 or 1.5; when the organ or tissue is the kidney, the mean SUVR threshold is approximately 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2 or 2.3; and the organ If the tissue is the pancreas, the mean SUVR threshold is approximately 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, or 1.7; if the organ or tissue is the heart, the mean SUVR threshold is approximately 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, or 2.1; or if the organ or tissue is the adrenal gland, the mean SUVR threshold is approximately 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, or 1.4. In some embodiments, the mean SUVR threshold is 1.31 when the organ or tissue is the liver, 1.21 when the organ or tissue is the spleen, 1.88 when the organ or tissue is the kidney, 1.40 when the organ or tissue is the pancreas, 1.19 when the organ or tissue is the heart, or 1.35 when the organ or tissue is the adrenal gland.

[0037] In some embodiments, the threshold is the SUVR maximum threshold. In some embodiments, when the organ or tissue is the liver, the SUVR maximum threshold is 10.6, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, or 16; when the organ or tissue is the spleen, the SUVR maximum threshold is 3.8, 4, 4.5, 5, 5.5, or 5.9; and when the organ or tissue is the kidney, the SUVR maximum threshold is 7.7, 8, 8.5, 9, 9.5, 10, 10.5. If the organ or tissue is the pancreas, the maximum SUVR threshold is 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, or 11.4 if the organ or tissue is the heart, or 1, 1.1, 1.2, 1.3, 1.4, 1.5, or 1.6 if the organ or tissue is the adrenal gland. In some embodiments, the maximum SUVR threshold is 13.26 when the organ or tissue is the liver, 4.86 when the organ or tissue is the spleen, 9.68 when the organ or tissue is the kidney, 9.48 when the organ or tissue is the pancreas, 6.20 when the organ or tissue is the heart, or 1.31 when the organ or tissue is the adrenal gland.

[0038] In some embodiments, if amyloid is detected in the heart, the method further includes performing a cardiac biopsy. In some embodiments, the method further includes performing additional amyloid imaging on the individual. In some embodiments, the method further includes performing additional amyloid imaging on the individual before administering an amyloid-reactive peptide. In some embodiments, the additional amyloid imaging on the individual includes ECHO, CMR, bone scintigraphy, or positron emission tomography imaging. In some embodiments, the additional amyloid imaging on the individual is 99mTc-PyP, 99m Tc-DPD, 99m Tc-HMDP, 99m Tc-MDP, other bone scintigraphy tracers, 124 I and 18 F-Florbetapyr, 18 F-Frutemtamol and 18 The present invention further includes a tracer selected from the group consisting of F-florbetabene.

[0039] In some embodiments, the diagnosis, prognosis, or response is confirmed by an indicator of the individual's health-related quality of life, the presence or amount of one or more biomarkers associated with amyloid-related disease, cardiac biopsy, and / or additional amyloid imaging of the individual.

[0040] In some embodiments, the individual is diagnosed with amyloid cardiomyopathy. In some embodiments, the individual is suspected of having amyloid cardiomyopathy.

[0041] In some embodiments, amyloid-related disease is systemic or focal amyloidosis. In some embodiments, amyloid-related disease is cardiac amyloidosis.

[0042] In some embodiments, the amyloid-reactive peptide has pan-amyloid specificity. In some embodiments, the amyloid-reactive peptide is immunoglobulin light chain (AL), immunoglobulin heavy chain (AH), β2 microglobulin (Aβ2M), transthyretin (ATTR wild type, ATTR variant), apolipoprotein AI (AApoAI), apolipoprotein AII (AApoAII), apolipoprotein AIV (AApoAIV), gelsolin (AGe1), apolipoprotein C-II (AApoCII), apolipoprotein C-II (AApoCIII), lysozyme It binds to amyloid proteins such as ALys, leukocyte chemotactic factor (ALECT2), fibrinogen A variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactoadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP), α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), IAAP, and ALβ4 or ALβ1.

[0043] In some embodiments, amyloid-reactive peptides are detected using PET, PET / CT, gamma machinography, SPECT, and / or SPECT / CT.

[0044] In some embodiments, amyloid-related disease is selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AApoCII, AApoCIII, AGel, Alys, ALECT2, AFib, ACys, ACa, AMed, AIAPP, APro, AIns, APrP, and Aβ amyloidosis. In some embodiments, the amyloid-related disease is cardiac AL amyloidosis. In some embodiments, the amyloid-related disease is cardiac ATTR amyloidosis.

[0045] In some embodiments, the individual has a genetic predisposition to amyloid-related disease. In some embodiments, the individual has a family history of amyloid-related disease.

[0046] In some embodiments, the individual is elderly.

[0047] In some embodiments, the individual has an early stage of amyloid-related disease. In some embodiments, the individual has an early stage of systemic amyloidosis. In some embodiments, the individual has an early stage of cardiac amyloidosis. In some embodiments, the individual has an early stage of AL amyloidosis. In some embodiments, the early stage of AL amyloidosis is diagnosed according to the Mayo Clinic system. In some embodiments, the early stage of AL amyloidosis is stage 1 AL amyloidosis. In some embodiments, the individual has an early stage of ATTR amyloidosis. In some embodiments, the early stage of ATTR amyloidosis includes stage 1 ATTR amyloidosis.

[0048] In some embodiments, the method further includes providing treatment for amyloid-related diseases. In some embodiments, the treatment for amyloid-related diseases is selected from the group comprising transthyretin stabilizers (e.g., tafamidis, acolamidis), transthyretin silencers (e.g., patisirane, inotercene, butricirane, eproetercene), gene editing approaches, anti-amyloid approaches using monoclonal antibodies, therapies targeting plasma cell clones (e.g., daratumumab, bortezomib), and antibody-peptide fusions comprising antibody-peptide fusion proteins containing an antibody that binds to amyloid fibril and a second amyloid-reactive peptide. In some embodiments, the second amyloid-reactive peptide comprises an amino acid sequence selected from the group comprising SEQ ID NOs: 1 to 14, including substitutions, insertions, or deletions of 0, 1, 2, 3, or 4 amino acids. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain and a light chain, where the amyloid-reactive peptide is bound to the C-terminus of the antibody's light chain via a spacer. In some embodiments, the antibody-peptide fusion protein comprises (i) an amyloid-reactive peptide having the amino acid sequence described in SEQ ID NO: 2, and (ii) an antibody that binds to human amyloid fibril, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy chain of the antibody comprises a heavy chain variable region (VH), and the light chain of the antibody comprises a light chain variable region (VL), wherein VH comprises CDR-H1 having the amino acid sequence described in SEQ ID NO: 106, CDR-H2 having the amino acid sequence described in SEQ ID NO: 107, and CDR-H3 having the amino acid sequence described in SEQ ID NO: 108, and VL comprises CDR-L1 having the amino acid sequence described in SEQ ID NO: 103, CDR-L2 having the amino acid sequence described in SEQ ID NO: 104, and CDR-L3 having the amino acid sequence described in SEQ ID NO: 105, wherein the amyloid-reactive peptide and the antibody are bound at the C-terminus of the light chain, wherein the amyloid-reactive peptide is bound to the antibody via a spacer having the amino acid sequence described in SEQ ID NO: 83.

[0049] In some embodiments, an organism is administered a peptide containing an amyloid-reactive peptide and an N-terminal leader sequence. In some embodiments, the peptide containing the amyloid-reactive peptide contains the amino acid sequence described in SEQ ID NO: 100.

[0050] In some embodiments, the detectable label is bound to one or more amino acids at positions 1-6 of the amino acid sequence described in SEQ ID NO: 100. In some embodiments, the detectable label is 124 I. In some embodiments, 124 I is attached to the amino acid at position 4 of sequence number 100.

[0051] In some embodiments, the method further includes administering potassium iodide to an individual. In some embodiments, the administration of potassium iodide includes administering 130 mg of potassium iodide to the patient over a period of 7 days, starting one day before the administration of the amyloid-reactive peptide. In some embodiments, the administration of potassium iodide includes administering 130 mg of potassium iodide to the patient over a period of 3 days, starting about 30 minutes before the administration of the amyloid-reactive peptide. In some embodiments, the administration of potassium iodide is by oral administration.

[0052] In another embodiment, this specification provides a peptide comprising the amino acids described in Sequence ID No. 100, wherein the peptide comprises a detectable label attached to one or more amino acids at positions 1-6. In some embodiments, the detectable label is 124 I. In some embodiments, 124 I is bonded to amino acid 4 of sequence number 100. [Brief explanation of the drawing]

[0053] The following drawings illustrate specific features and advantages of the present disclosure. These embodiments are not intended to limit the scope of the appended claims in any way. [Figure 1-1]Figure 1 shows a multipart, open-label study to evaluate the safety, dosemetry, and biodistribution of the 124I-p5+14 peptide in patients with any type of systemic amyloidosis, pre-symptomatic subjects with germline TTR mutations, and healthy subjects. [Figure 1-2] Figure 1 shows a multipart, open-label study to evaluate the safety, dosemetry, and biodistribution of the 124I-p5+14 peptide in patients with any type of systemic amyloidosis, pre-symptomatic subjects with germline TTR mutations, and healthy subjects. [Figure 2A] Figure 2 shows the in vivo distribution of radioactivity in healthy subjects. Figure 2A shows the maximum intensity projection of PET data obtained 5 hours after injection for five healthy subjects. Figure 2B shows a transverse PET / CT image of the heart of a healthy subject. [Figure 2B] Figure 2 shows the in vivo distribution of radioactivity in healthy subjects. Figure 2A shows the maximum intensity projection of PET data obtained 5 hours after injection for five healthy subjects. Figure 2B shows a transverse PET / CT image of the heart of a healthy subject. [Figure 3A] Figure 3 shows cardiac uptake of 124I-p5+14 in patients with AL and ATTR-associated amyloidosis. Figure 3A shows the maximum intensity projection of PET data and cardiac coronal PET / CT images (image intensity is individually set for optimal visualization). Figure 3B shows transverse cardiac PET / CT images in patients with AL or ATTR amyloidosis and healthy subjects with positive PET imaging (PET+) considered positive (Clin+) or negative (Clin-) for cardiac amyloidosis in medical records. [Figure 3B]Figure 3 shows cardiac uptake of 124I-p5+14 in patients with AL and ATTR-associated amyloidosis. Figure 3A shows the maximum intensity projection of PET data and cardiac coronal PET / CT images (image intensity is individually set for optimal visualization). Figure 3B shows transverse cardiac PET / CT images in patients with AL or ATTR amyloidosis and healthy subjects with positive PET imaging (PET+) considered positive (Clin+) or negative (Clin-) for cardiac amyloidosis in medical records. [Figure 4A-1] Figure 4 shows the PET evaluation of the cardiac 124I-p5+14 SUVR mean (SUVRmean) and its correlation with serum NTproBNP. Figure 4A shows a comparison of cardiac SUVR mean for patients with AL, ATTR, other types of amyloidosis, and healthy subjects. Cardiac SUVR mean was also compared for PET-positive and PET-negative AL and ATTR patients. Figures 4B-4C show a significant positive correlation between cardiac SUVR mean and serum NTproBNP in AL patients (Figure 4B), which was not significant in ATTR patients (Figure 4C). Figure 4D shows the correlation between serum NTproBNP in AL patients and cardiac SUVR mean scaled by serum creatinine for each patient. [Figure 4A-2] Figure 4 shows the PET evaluation of the cardiac 124I-p5+14 SUVR mean (SUVRmean) and its correlation with serum NTproBNP. Figure 4A shows a comparison of cardiac SUVR mean for patients with AL, ATTR, other types of amyloidosis, and healthy subjects. Cardiac SUVR mean was also compared for PET-positive and PET-negative AL and ATTR patients. Figures 4B-4C show a significant positive correlation between cardiac SUVR mean and serum NTproBNP in AL patients (Figure 4B), which was not significant in ATTR patients (Figure 4C). Figure 4D shows the correlation between serum NTproBNP in AL patients and cardiac SUVR mean scaled by serum creatinine for each patient. [Figure 4B]Figure 4 shows the PET evaluation of the cardiac 124I-p5+14 SUVR mean (SUVRmean) and its correlation with serum NTproBNP. Figure 4A shows a comparison of cardiac SUVR mean for patients with AL, ATTR, other types of amyloidosis, and healthy subjects. Cardiac SUVR mean was also compared for PET-positive and PET-negative AL and ATTR patients. Figures 4B-4C show a significant positive correlation between cardiac SUVR mean and serum NTproBNP in AL patients (Figure 4B), which was not significant in ATTR patients (Figure 4C). Figure 4D shows the correlation between serum NTproBNP in AL patients and cardiac SUVR mean scaled by serum creatinine for each patient. [Figure 4C] Figure 4 shows the PET evaluation of the cardiac 124I-p5+14 SUVR mean (SUVRmean) and its correlation with serum NTproBNP. Figure 4A shows a comparison of cardiac SUVR mean for patients with AL, ATTR, other types of amyloidosis, and healthy subjects. Cardiac SUVR mean was also compared for PET-positive and PET-negative AL and ATTR patients. Figures 4B-4C show a significant positive correlation between cardiac SUVR mean and serum NTproBNP in AL patients (Figure 4B), which was not significant in ATTR patients (Figure 4C). Figure 4D shows the correlation between serum NTproBNP in AL patients and cardiac SUVR mean scaled by serum creatinine for each patient. [Figure 4D]Figure 4 shows the PET evaluation of the cardiac 124I-p5+14 SUVR mean (SUVRmean) and its correlation with serum NTproBNP. Figure 4A shows a comparison of cardiac SUVR mean for patients with AL, ATTR, other types of amyloidosis, and healthy subjects. Cardiac SUVR mean was also compared for PET-positive and PET-negative AL and ATTR patients. Figures 4B-4C show a significant positive correlation between cardiac SUVR mean and serum NTproBNP in AL patients (Figure 4B), which was not significant in ATTR patients (Figure 4C). Figure 4D shows the correlation between serum NTproBNP in AL patients and cardiac SUVR mean scaled by serum creatinine for each patient. [Figure 5A] Figures 5A-5B show extracardiac uptake of 124I-p5+14 in patients with AL or ATTR-associated amyloidosis. Uptake of 124I-p5+14 in the lungs, fat, kidneys, pancreas, joints, and spine of patients with ATTR amyloidosis (Figure 5A), and in the liver, spleen, kidneys, adrenal glands, bone lesions, choroid plexus, pituitary gland, bone marrow, and pancreas of patients with AL amyloidosis and multiple myeloma and amyloidosis (Figure 5B). [Figure 5B] Figures 5A-5B show extracardiac uptake of 124I-p5+14 in patients with AL or ATTR-associated amyloidosis. Uptake of 124I-p5+14 in the lungs, fat, kidneys, pancreas, joints, and spine of patients with ATTR amyloidosis (Figure 5A), and in the liver, spleen, kidneys, adrenal glands, bone lesions, choroid plexus, pituitary gland, bone marrow, and pancreas of patients with AL amyloidosis and multiple myeloma and amyloidosis (Figure 5B). [Figure 6] Figure 6 shows automated contour plotting on a 124I-p5+14 PET image for quantifying cardiac uptake of a radioactive tracer. LA: left atrium. LV: left ventricle. PET / CT: positron emission tomography / computed tomography. RV: right ventricle. SD: standard deviation. VOI: volume of interest. %ID / mL: percentage of injected volume per milliliter. [Figure 7]Figure 7 shows 124I-p5+14 PET / CT in uptake of an ATTRwt cardiomyopathy (CMP) patient with an indistinct 99mPYP SPECT / CT. The individual presented with heart failure, preserved left ventricular ejection fraction, grade 2 diastolic dysfunction, asymmetric cardiac hypertrophy, and overall longitudinal strain impairment on echocardiography. He had a history of surgery for bilateral carpal tunnel syndrome and lumbar spinal stenosis and no evidence of abnormal immunoglobulin light chains. The 99mTc-PYP SPECT / CT was indistinct (undiagnostic) with grade 1 uptake. Cardiac MRI subsequently showed diffuse gadolinium delayed enhancement. Endocardial biopsy using normal TTR gene sequencing and mass spectrometry confirmed ATTRwt-CMP (NAC stage 2, Mayo stage 2). 124I-p5+14 PET / CT performed within 6 months showed clear myocardial uptake. NAC: National Amyloidosis Center. MRI: Magnetic Resonance Imaging. 99mTc-PYP SPECT / CT: 99mTc-Pyrophosphate Single-Photon Emission Computed Tomography / Computed Tomography. [Figure 8-1] Figure 8 shows the uptake metrics for 124I-p5+14. 124I-p5+14 LV %ID, CAA, and TBR completely distinguished cases of amyloid CMP from controls, but the mean SUV did not. Measurement and calculation of uptake are described in the "Methods" section. Thresholds shown for high metrics were defined using the Youden index. Intergroup comparisons were performed using Dunn's test. CAA: Cardiac amyloid activity (SUV units × mL). LV: Left ventricle. SUV: Standardized uptake value. TBR: Target vs. background ratio. %ID: Percentage of injected volume. [Figure 8-2]Figure 8 shows the uptake metrics for 124I-p5+14. 124I-p5+14 LV %ID, CAA, and TBR completely distinguished cases of amyloid CMP from controls, but the mean SUV did not. Measurement and calculation of uptake are described in the "Methods" section. Thresholds shown for high metrics were defined using the Youden index. Intergroup comparisons were performed using Dunn's test. CAA: Cardiac amyloid activity (SUV units × mL). LV: Left ventricle. SUV: Standardized uptake value. TBR: Target vs. background ratio. %ID: Percentage of injected volume. [Figure 9-1] Figure 9 shows 18F-florbetapir uptake metrics. 18F-florbetapir LV %ID and CAA completely differentiated cases of amyloid CMP from controls, but mean SUV and TBR did not. The thresholds shown for high metrics were defined using the Youden index. Intergroup comparisons were performed using Dunn's test. [Figure 9-2] Figure 9 shows 18F-florbetapir uptake metrics. 18F-florbetapir LV %ID and CAA completely differentiated cases of amyloid CMP from controls, but mean SUV and TBR did not. The thresholds shown for high metrics were defined using the Youden index. Intergroup comparisons were performed using Dunn's test. [Figure 10-1] Figure 10 shows the uptake metrics for 124I-p5+14 and 18F-florbetapil in the same subjects. Imaging for 124I-p5+14 and 18F-florbetapil was obtained in the same subjects (one AL-CMP subject without 18F-florbetapil PET / CT results is not included). When 124I-p5+14 was used, LV %ID, mean SUV, and CAA were higher in ATTRwt-CMP. However, when 18F-florbetapil was used in AL-CMP and ATTRwt-CMP, LV TBR was higher. The measurement and calculation of uptake are described in the "Methods" section. P-values ​​were obtained from paired Wilcoxon signed-rank tests. [Figure 10-2] Figure 10 shows the uptake metrics for 124I-p5+14 and 18F-florbetapil in the same subjects. Imaging for 124I-p5+14 and 18F-florbetapil was obtained in the same subjects (one AL-CMP subject without 18F-florbetapil PET / CT results is not included). When 124I-p5+14 was used, LV %ID, mean SUV, and CAA were higher in ATTRwt-CMP. However, when 18F-florbetapil was used in AL-CMP and ATTRwt-CMP, LV TBR was higher. The measurement and calculation of uptake are described in the "Methods" section. P-values ​​were obtained from paired Wilcoxon signed-rank tests. [Figure 11A] Figures 11A and 11B show the changes in 124I-p5+14 uptake, measured by SUVR mean and NTproBNP, in patients with amyloidosis AL (Figure 11A) and ATTR (Figure 11B). [Figure 11B] Figures 11A and 11B show the changes in 124I-p5+14 uptake, measured by SUVR mean and NTproBNP, in patients with amyloidosis AL (Figure 11A) and ATTR (Figure 11B). [Figure 12] Figure 12 shows the relationship between the percentage change in NTproBNP and the scaled percentage change in cardiac SUVR in 19 amyloidosis patients with AL or ATTR. [Figure 13A]Figures 13A–13I show the relationships between anatomical and functional metrics related to cardiac 124I-p5+14 uptake, measured by mean SUVR, for AL patients, ATTR patients, and all amyloidosis patients. In AL patients, cardiac left ventricular wall thickness (LV) (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13A). Overall longitudinal strain (GLS) (Y-axis), a parameter reflecting left ventricular function, correlates with mean cardiac SUVR (X-axis) in AL patients (Figure 13B). In AL patients, interventricular septal thickness (IVS) (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13C). In ATTR patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13D). In ATTR patients, GLS (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13E). In ATTR patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13F). In amyloidosis patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13G). In amyloidosis patients, GLS (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13H). In amyloidosis patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13I). [Figure 13B]Figures 13A–13I show the relationships between anatomical and functional metrics related to cardiac 124I-p5+14 uptake, measured by mean SUVR, for AL patients, ATTR patients, and all amyloidosis patients. In AL patients, cardiac left ventricular wall thickness (LV) (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13A). Overall longitudinal strain (GLS) (Y-axis), a parameter reflecting left ventricular function, correlates with mean cardiac SUVR (X-axis) in AL patients (Figure 13B). In AL patients, interventricular septal thickness (IVS) (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13C). In ATTR patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13D). In ATTR patients, GLS (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13E). In ATTR patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13F). In amyloidosis patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13G). In amyloidosis patients, GLS (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13H). In amyloidosis patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13I). [Figure 13C]Figures 13A–13I show the relationships between anatomical and functional metrics related to cardiac 124I-p5+14 uptake, measured by mean SUVR, for AL patients, ATTR patients, and all amyloidosis patients. In AL patients, cardiac left ventricular wall thickness (LV) (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13A). Overall longitudinal strain (GLS) (Y-axis), a parameter reflecting left ventricular function, correlates with mean cardiac SUVR (X-axis) in AL patients (Figure 13B). In AL patients, interventricular septal thickness (IVS) (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13C). In ATTR patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13D). In ATTR patients, GLS (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13E). In ATTR patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13F). In amyloidosis patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13G). In amyloidosis patients, GLS (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13H). In amyloidosis patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13I). [Figure 13D]Figures 13A–13I show the relationships between anatomical and functional metrics related to cardiac 124I-p5+14 uptake, measured by mean SUVR, for AL patients, ATTR patients, and all amyloidosis patients. In AL patients, cardiac left ventricular wall thickness (LV) (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13A). Overall longitudinal strain (GLS) (Y-axis), a parameter reflecting left ventricular function, correlates with mean cardiac SUVR (X-axis) in AL patients (Figure 13B). In AL patients, interventricular septal thickness (IVS) (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13C). In ATTR patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13D). In ATTR patients, GLS (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13E). In ATTR patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13F). In amyloidosis patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13G). In amyloidosis patients, GLS (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13H). In amyloidosis patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13I). [Figure 13E]Figures 13A–13I show the relationships between anatomical and functional metrics related to cardiac 124I-p5+14 uptake, measured by mean SUVR, for AL patients, ATTR patients, and all amyloidosis patients. In AL patients, cardiac left ventricular wall thickness (LV) (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13A). Overall longitudinal strain (GLS) (Y-axis), a parameter reflecting left ventricular function, correlates with mean cardiac SUVR (X-axis) in AL patients (Figure 13B). In AL patients, interventricular septal thickness (IVS) (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13C). In ATTR patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13D). In ATTR patients, GLS (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13E). In ATTR patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13F). In amyloidosis patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13G). In amyloidosis patients, GLS (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13H). In amyloidosis patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13I). [Figure 13F]Figures 13A–13I show the relationships between anatomical and functional metrics related to cardiac 124I-p5+14 uptake, measured by mean SUVR, for AL patients, ATTR patients, and all amyloidosis patients. In AL patients, cardiac left ventricular wall thickness (LV) (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13A). Overall longitudinal strain (GLS) (Y-axis), a parameter reflecting left ventricular function, correlates with mean cardiac SUVR (X-axis) in AL patients (Figure 13B). In AL patients, interventricular septal thickness (IVS) (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13C). In ATTR patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13D). In ATTR patients, GLS (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13E). In ATTR patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13F). In amyloidosis patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13G). In amyloidosis patients, GLS (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13H). In amyloidosis patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13I). [Figure 13G]Figures 13A–13I show the relationships between anatomical and functional metrics related to cardiac 124I-p5+14 uptake, measured by mean SUVR, for AL patients, ATTR patients, and all amyloidosis patients. In AL patients, cardiac left ventricular wall thickness (LV) (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13A). Overall longitudinal strain (GLS) (Y-axis), a parameter reflecting left ventricular function, correlates with mean cardiac SUVR (X-axis) in AL patients (Figure 13B). In AL patients, interventricular septal thickness (IVS) (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13C). In ATTR patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13D). In ATTR patients, GLS (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13E). In ATTR patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13F). In amyloidosis patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13G). In amyloidosis patients, GLS (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13H). In amyloidosis patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13I). [Figure 13H]Figures 13A–13I show the relationships between anatomical and functional metrics related to cardiac 124I-p5+14 uptake, measured by mean SUVR, for AL patients, ATTR patients, and all amyloidosis patients. In AL patients, cardiac left ventricular wall thickness (LV) (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13A). Overall longitudinal strain (GLS) (Y-axis), a parameter reflecting left ventricular function, correlates with mean cardiac SUVR (X-axis) in AL patients (Figure 13B). In AL patients, interventricular septal thickness (IVS) (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13C). In ATTR patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13D). In ATTR patients, GLS (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13E). In ATTR patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13F). In amyloidosis patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13G). In amyloidosis patients, GLS (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13H). In amyloidosis patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13I). [Figure 13I]Figures 13A–13I show the relationships between anatomical and functional metrics related to cardiac 124I-p5+14 uptake, measured by mean SUVR, for AL patients, ATTR patients, and all amyloidosis patients. In AL patients, cardiac left ventricular wall thickness (LV) (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13A). Overall longitudinal strain (GLS) (Y-axis), a parameter reflecting left ventricular function, correlates with mean cardiac SUVR (X-axis) in AL patients (Figure 13B). In AL patients, interventricular septal thickness (IVS) (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13C). In ATTR patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13D). In ATTR patients, GLS (Y-axis) does not correlate with mean cardiac SUVR (X-axis) (Figure 13E). In ATTR patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13F). In amyloidosis patients, cardiac LV wall thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13G). In amyloidosis patients, GLS (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13H). In amyloidosis patients, IVS thickness (Y-axis) correlates with mean cardiac SUVR (X-axis) (Figure 13I).

[0054] Detailed explanation All publications, including patent documents, scientific articles, and databases, referenced in this application are incorporated herein by reference for all purposes, just as each individual publication is incorporated herein by reference individually. In the event that any definition contained herein contradicts or is inconsistent with any definition contained in any patent, application, published application, or other publication incorporated herein by reference, the definition contained herein shall prevail over the definition incorporated herein by reference.

[0055] The section headings used in this specification are for organizational purposes only and should not be interpreted as limiting the subjects discussed.

[0056] I. Overview This specification provides methods for the diagnosis and treatment of amyloid-related diseases. In some embodiments, the method includes administering an amyloid-reactive substance and detecting the amount of amyloid-reactive peptide by detecting the amount of a detectable label in the tissues or organs of an individual. In some embodiments, the method provided herein can diagnose amyloid-related disease in an individual at risk of developing amyloid-related disease. In some embodiments, the method provided herein can diagnose amyloid-related disease in an individual suspected of having amyloid-related disease. In some embodiments, the method provided herein can determine the prognosis of an individual diagnosed with amyloid-related disease. In some embodiments, the method provided herein can monitor the treatment of an individual having amyloid-related disease. In some embodiments, the method provided herein can select a treatment for an individual having amyloid-related disease.

[0057] The premature mortality rate in newly diagnosed amyloid-related diseases suggests that significant delays in diagnosis still exist (Muchtar E, Gertz MA, Kumar SK et al., Blood. 2017;129(15):2111-2119). For example, approximately 20% of patients with AL amyloidosis die within six months of diagnosis, and this statistic has not improved at all over the past 40 years, suggesting that despite significant advances in the treatment of this disease, patients diagnosed at an advanced stage may not be saved (Hasib Sidiqi, M and Morie A Gertz. Blood cancer journal vol.11,5 90.15 May.2021). Once a diagnosis of amyloidosis is confirmed, current anatomical imaging techniques, including echocardiography and magnetic resonance imaging, provide valuable structural and functional information. 99mWhile Tc-PyP and similar osteoaffinic substances are used to detect cardiac amyloid, their relationship to amyloid fibrils is not fully understood. Therefore, non-invasive imaging techniques using reagents that directly bind to amyloid deposits, are quantifiable, and can detect cardiac amyloidosis before the onset of apparent anatomical changes and organ dysfunction would offer significant clinical benefits.

[0058] In some embodiments, this specification provides a method for diagnosing amyloid-related disease in an individual at risk of developing amyloid-related disease, the method comprising: administering to the individual an amyloid-reactive peptide comprising an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label; and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the tissue or organ of the individual, wherein an amount of the amyloid-reactive peptide exceeding a threshold indicates that the individual has amyloid-related disease.

[0059] Furthermore, in some embodiments herein, methods for treating amyloid-related diseases are provided, the methods comprising: administering an amyloid-reactive peptide to an individual, comprising an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label; detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs; and, if the amount of the amyloid-reactive peptide exceeds a threshold, performing treatment for the amyloid-related disease.

[0060] II. Definition As used herein, “amino acid” or “amino acid residue” means any naturally occurring amino acid, any non-naturally occurring amino acid, any modified amino acid (including derivatized amino acids), or any amino acid mimetic known in the art. Amino acids may be represented by both their common three-letter abbreviations and one-letter abbreviations.

[0061] The terms amyloid, amyloid deposition (amyloid deposits), amyloid fibrils, and amyloid fibers refer to insoluble fibrous protein aggregates that share specific structural characteristics. Protein aggregates are, for example, tertiary structures formed by the aggregation of any one of several different proteins, and have a tertiary structure consisting of a regular arrangement of β-sheets stacked perpendicular to the fiber axis. See Sunde et al., J.Mol.Biol. (1997) 273:729-39. Abnormal accumulation of amyloid in organs can lead to amyloidosis. Although diverse in development, all amyloids share the following common morphological characteristics: they stain with certain dyes such as Congo red and, after staining, exhibit characteristic red-green birefringence under polarized light. Amyloids also share common ultrastructural features as well as common X-ray diffraction and infrared spectra.

[0062] Amyloidosis refers to a condition or disease characterized by the presence of amyloid, such as amyloid deposits. "Amyloid disease" or "amyloidosis" is a disease associated with the formation, deposition, accumulation, or persistence of amyloid fibrils. Such diseases include, but are not limited to, Alzheimer's disease, Down syndrome, hereditary cerebral hemorrhage with Dutch-type amyloidosis, and cerebral beta-amyloid angiopathy. Other amyloid diseases, such as systemic AA amyloidosis, AL amyloidosis, ATTR amyloidosis, ALECT2 amyloidosis, and IAPP amyloidosis in type II diabetes, are also amyloid diseases.

[0063] The term "amyloidogenic" refers to the formation or tendency to form amyloid deposits. For example, certain soluble monomeric proteins undergo extensive conformational changes, resulting in aggregation into well-ordered, unbranched fibrils 8-10 nm wide, which ultimately form amyloid aggregates. In humans, for instance, more than 30 proteins have been found to form amyloid deposits (or amyloid). Not all proteins within a diverse class of proteins, such as immunoglobulin light chains, can form amyloid. That is, some immunoglobulin light chains are amyloid-non-amyloid, meaning they do not have a tendency to form amyloid. However, other proteins in the same class can form amyloid deposits and are therefore amyloidogenic. Furthermore, some within a class of light chain proteins can be considered more "amyloidogenic" than others based on their ease of amyloid fibril formation. Certain light chain proteins are considered amyloid-non-amyloid or amyloid-hypoplastic because they do not readily form amyloid fibrils in patients or in vitro.

[0064] As used herein, the term “carrier” includes pharmaceutically acceptable carriers, excipients, or stabilizers that are nontoxic to cells, tissues, mammals, or subjects to which they are exposed at the doses and concentrations used. In many cases, pharmaceutically acceptable carriers are aqueous pH buffer solutions. Examples of pharmaceutically acceptable carriers include, but are not limited to, buffers such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid; low molecular weight (less than approximately 10 residues) polypeptides; proteins, e.g., serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, e.g., glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, such as EDTA; sugar alcohols, such as mannitol or sorbitol; salt-forming counterions, such as sodium; and / or nonionic surfactants, such as Tween®, polyethylene glycol (PEG), and Pluronics®.

[0065] As used herein, the terms “effective dose” or “appropriate dose” refer to an amount sufficient to produce a beneficial or desired clinical or biochemical outcome. An effective dose may be administered once or multiple times. For the purposes of this invention, an effective dose of amyloid-reactive material or detection is an amount sufficient to bind to amyloid and enable the detection of amyloid.

[0066] As used herein, the interchangeable terms “imaging agent” or “contrast agent” mean any substance that can be used in connection with a method for imaging an internal region of an object and / or diagnosing the presence or absence of disease in an object by application and / or detection of an energy source. Exemplary imaging agents include contrast agents used in connection with ultrasound, magnetic resonance imaging, radionuclide imaging or X-ray (including computed tomography) imaging of a patient, and the compositions described herein.

[0067] As used herein, the term “mammal” in the context of the present invention means any animal classified as a mammal, including humans, livestock and farm animals, and zoo animals, sports animals, or pet animals, such as dogs, cats, cows, horses, sheep, and pigs. In some embodiments, the mammal is a human.

[0068] As used herein, the term "peptide" means any peptide or peptide mimetic structure comprising two or more amino acids, including chemical modifications and derivatives of amino acids.

[0069] As used herein, the terms “purified” or “isolated” molecules mean biological or synthetic molecules that have been taken out of their natural environment, isolated or separated, and that do not contain other naturally associated components.

[0070] As used herein, the term "specifically binds" refers to a non-random binding reaction between two molecules, for example, between an amyloid-reactive substance and amyloid. The term "specifically binds" may be used interchangeably with "selectively target" or "selectively associate."

[0071] As used herein, the terms “selectively targeting” or “selectively associating” with respect to amyloid mean, for example, selective localization or binding between an amyloid-reactive substance and amyloid, compared to the case of a non-amyloid protein. Amyloid-reactive substances may selectively target multiple types of amyloid.

[0072] As used herein, the term "subject" means a vertebrate. This vertebrate may be a mammal, such as a human. The subject may be a human patient.

[0073] As used herein, the term "amyloid-reactive substance" refers to a substance that specifically reacts with or specifically binds to amyloid.

[0074] As used herein, the term "conjugate" includes both covalent and non-covalent bonds.

[0075] In this specification, the term "antibody" is used in its broadest sense and particularly includes monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, insofar as they exhibit the desired biological activity.

[0076] The term "amyloid-reactive peptide" refers to a peptide that binds to any of the following amyloid fibrils: amyloid-forming λ6 variable region protein (Vλ6Wil) or amyloid-forming immunoglobulin light chain (AL), Aβ(1-40) amyloid-like fibrils or amyloid-forming AP precursor protein, or serum amyloid protein A (AA). In other embodiments, the amyloid bound by the amyloid-reactive peptide is immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin variant (ATTRv), wild-type transthyretin (ATTRwt), apolipoprotein AI (AApoAI), apolipoprotein AII (AApoAII), gelzolin (AGel), lysozyme (ALys), leukocyte chemotactic factor (ALect2), fibrinogen. This includes amyloid-forming forms of α-synucine variants (AFib), cystatin variants (ACys), calcitonin (ACal), lactadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP); α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), or other amyloid-forming peptides such as IAAP, ALκ, and ALλ1.

[0077] II. Diagnostic Methods This specification provides a method for diagnosing amyloid-related disease in an individual at risk of developing amyloid-related disease, the method comprising: administering to the individual an amyloid-reactive peptide comprising an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label; and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissue or organ, wherein an amount of the amyloid-reactive peptide exceeding a threshold indicates that the individual has amyloid disease.

[0078] This specification provides a method for diagnosing an early stage of amyloid-related disease in an individual, the method comprising administering an amyloid-reactive peptide conjugated to a detectable label to the individual, and detecting the amount of the detectable label in the individual's tissues or organs, where an amount of the amyloid-reactive peptide exceeding a threshold indicates that the individual has amyloid-related disease, and the amyloid-related disease is an early stage of amyloid-related disease. In some embodiments, the amyloid-related disease is systemic amyloidosis. In some embodiments, the amyloid-related disease is cardiac amyloidosis. In some embodiments, the amyloid-reactive peptide conjugated to a detectable label is 124 It is a peptide containing the amino acids listed in SEQ ID NO: 100, conjugated to I. In some embodiments, 124 I is bonded to amino acid 4 of sequence number 100.

[0079] The methods provided herein can be used to diagnose early-stage amyloid-related diseases. In some embodiments, the methods can be used to diagnose amyloid-related diseases before conventional diagnostic methods. In some embodiments, the individual has been previously diagnosed as not having amyloid-related disease. In some embodiments, the methods have high sensitivity and specificity for diagnosing early-stage amyloid-related diseases.

[0080] A. Amyloid-reactive peptides In some embodiments, a method for diagnosing a type of amyloid disease includes administering an amyloid-reactive peptide conjugated to a detectable label.

[0081] In some embodiments, a method for diagnosing a type of amyloid disease includes administering an amyloid-reactive substance comprising an amyloid-reactive peptide conjugated to a detectable label. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence that is at least 80%, 85%, 90%, or more identical to the amino acid sequence described as any one of SEQ ID NOs: 1 to 14, for example, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence described as any one of SEQ ID NOs: 1 to 14. In some embodiments, the amyloid-reactive peptide used in conjunction with the methods described herein comprises or consists of about 10 to 55 amino acids. The amyloid-reactive peptides of the present invention include, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 amino acids. Such peptides are described, for example, in International Patent Application WO2016032949 (which is incorporated herein by reference in its entirety). In some embodiments, a method for diagnosing amyloid disease includes an amyloid-reactive peptide having the amino acid sequence described in Sequence ID No. 13. In some embodiments, a method for diagnosing the type of amyloid disease includes p5+14. In some embodiments, a method for diagnosing amyloid disease is 124 It contains an amyloid-reactive peptide having the amino acid sequence described in SEQ ID NO: 13, conjugated to I. [Table 1]

[0082] The amino acids that form all or part of the amyloid-reactive peptide used in conjunction with this method may include stereoisomers and modifications of naturally occurring amino acids, amino acids not found in nature, post-translationally modified amino acids, enzymatically synthesized amino acids, derivatized amino acids, constructs or structures designed to mimic amino acids, and so on. The amino acids that form the peptide of the present invention may be one or more of the 20 common amino acids found in naturally occurring proteins, or one or more modified unusual amino acids. The amyloid-reactive peptide used in conjunction with the method described herein may be produced by any technique known to those skilled in the art, including chemical synthesis or recombination methods using standard molecular biological techniques.

[0083] The peptide of the present invention may also contain one or more modified amino acids. The modified amino acids may be derivatized amino acids or modified abnormal amino acids. Examples of modified abnormal amino acids include 2-aminoadipic acid (Aad), 3-aminoadipic acid (Baad), β-amino-propionic acid (Bala, β-alanine), 2-aminobutyric acid (Abu, piperidine acid), 4-aminobutyric acid (4Abu), 6-aminocaproic acid (Acp), 2-aminoheptanoic acid (Ahe), 2-aminoisobutyric acid (Aib), 3-aminoisobutyric acid (Baib), 2-aminopimelic acid (Apm), 2,4-diaminobutyric acid (Dbu), desmosine (Des), 2,2'-diaminopimelic acid (Dpm), 2,3-diaminopropionic acid (Dpr), and N-ethylglycerides. This includes, but is not limited to, lysine (EtGly), N-ethylasparagine (EtAsn), hydroxylysine (Hyl), allo-hydroxylysine (AHyl), 3-hydroxyproline (3Hyp), 4-hydroxyproline (4Hyp), isodesmosine (Ide), allo-isoleucine (Alle), N-methylglycine (MeGly, sarcosine), N-methylisoleucine (Melle), 6-N-methyllysine (MeLys), N-methylvaline (MeVal), norvaline (Nva), norleucine (Nle), and ornithine (Orn).

[0084] Other examples of modified abnormal amino acids are generally described in Synthetic Peptides: A User's Guide, Second Edition, April 2002, edited by Gregory A. Grant, Oxford University Press; Hruby VJ, Al-obeidi F and Kazmierski W: Biochem J 268:249-262, 1990; and Toniolo C: Int J Peptide Protein Res 35:287-300, 1990 (all of their teachings are incorporated herein by reference).

[0085] In certain embodiments, the peptide of the present invention may comprise, or consist of, the following amino acid sequence: SRAQRAQARQARQAQRAQRAQARQARQ (SEQ ID NO: 17).

[0086] The peptide of the present invention may be a fusion protein comprising a second peptide, for example, CGGY (SEQ ID NO: 18) or GGGYS (SEQ ID NO: 19), as a leader sequence at the amino terminus for labeling with a detection substance. Therefore, in some embodiments, the amyloid-reactive peptide may have up to 55 amino acids and may include the amino acid sequence described in CGGYSRAQRAQARQARQAQRAQRAQARQARQ (SEQ ID NO: 20).

[0087] In some embodiments, this specification provides peptides comprising an amyloid-reactive peptide fused to an N-terminal leader sequence. In some embodiments, the leader sequence comprises the amino acid sequence GGGYS (SEQ ID NO: 19). In some embodiments, this specification provides peptides comprising an amyloid-reactive peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 14 and the N-terminal leader sequence GGGYS. In some embodiments, this specification provides peptides comprising the amino acid sequence GGGYS KAQKA QAKQA KQAQK AQKAQ AKQAK QAQKA QKAQA KQAKQ (SEQ ID NO: 100). In some embodiments, the peptide comprises the amino acid sequence described in SEQ ID NO: 100 and is conjugated to a detectable label. In some embodiments, the detectable label is bound to one or more amino acids 1 to 6 of SEQ ID NO: 100. In some embodiments, the detectable label is bound to amino acid 4 of SEQ ID NO: 100. In some embodiments, the detectable label is bound to the N-terminal leader sequence. In some embodiments, the peptide comprises the amino acid sequence described in SEQ ID NO: 100. 124 I is conjugated to a detectable label. In some embodiments, 124I. The detectable label is bound to one or more amino acids 1-6 of SEQ ID NO: 100. In some embodiments, 124 The detectable label is bound to amino acid 4 of SEQ ID NO: 100. In some embodiments, the detectable label is bound to the N-terminus.

[0088] The fusion protein may contain other leader sequences, such as a cell-permeable peptide (CPP) or a blood-brain barrier (BBB) ​​crossing peptide.

[0089] The present invention also provides other peptides and fusion proteins rich in positively charged amino acids for imaging amyloid.

[0090] The peptides of the present invention can be produced by any technique known to those skilled in the art, including chemical synthesis, recombinant methods using standard molecular biological techniques, or isolation of peptides from natural sources. The peptides can be synthesized in solution or on a solid support according to conventional methods. Various automated synthesizers are commercially available and can be used according to known protocols [see, for example, Stewart and Young, Solid Phase Peptide Synthesis, 2nd ed. Pierce Chemical Co., 1984; Tam et al., J. Am. Chem. Soc., 105:6442, 1983; Merrifield, Science, 232:341-347, 1986; and Barany and Merrifield, The Peptides, edited by Gross and Meienhofer, Academic Press, New York, pp.1-284, 1979 (each in its entirety incorporated herein by reference)].

[0091] Alternatively, recombinant DNA technology can be employed, in which case the nucleotide sequence encoding the peptide of the present invention is inserted into an expression vector, transformed or transfected into a suitable host cell, cultured under conditions suitable for expression, and the peptide is isolated.

[0092] In certain embodiments, the amyloid-reactive material may be a naturally occurring peptide, which can be obtained by isolation or purification from its natural source. Protein purification techniques, to some extent, involve homogenization and crude fractionation of cells, tissues, or organs into peptide and non-peptide fractions. Other protein purification techniques include, for example, precipitation or thermal denaturation with ammonium sulfate, polyethylene glycol (PEG), antibodies, etc., followed by centrifugation; chromatographic steps such as ion exchange, gel filtration, reversed phase, hydroxyapatite, and affinity chromatography; isoelectric focusing; gel electrophoresis, such as polyacrylamide gel electrophoresis; and combinations of these and other techniques.

[0093] Various chromatographic techniques include, but are not limited to, ion exchange chromatography, gel exclusion chromatography, affinity chromatography, immunoaffinity chromatography, and reversed-phase chromatography. Particularly efficient methods for purifying peptides include medium- and low-pressure liquid chromatography (FPLC), or even high-performance liquid chromatography (HPLC).

[0094] The order in which the various purification steps are performed can be changed, or certain steps can be omitted, yet the method remains suitable for the preparation of substantially purified peptides. The peptides of the present invention may be part of a polypeptide or a protein, and may be produced by biochemical or enzymatic fragmentation of a polypeptide or protein. Thus, the peptides of the present invention may (a) be naturally occurring, (b) be produced by chemical synthesis, (c) be produced by recombinant DNA technology, (d) be produced by biochemical or enzymatic fragmentation of a larger molecule, (e) be produced by a method obtained from a combination of the methods a to d listed above, or (f) be produced by any other means for producing peptides.

[0095] In chemical synthesis, peptides can be modified at their N-terminus or C-terminus, resulting in improved stability and formulation, as well as resistance to protease degradation. Examples of amino acid modifications include pezylation, acetylation, alkylation, formylation, and amidation. Furthermore, various amino acids not naturally occurring can be introduced along the chain to improve peptide stability.

[0096] Cysteine ​​is also useful for promoting the labeling of the peptides of the present invention with biotin, fluorophores, or other ligands via conjugation. Furthermore, cysteine ​​on the leader peptide enables the generation of covalently bonded dimer molecules, which can increase the relative affinity of the peptide to the target.

[0097] B. Detectable labels In some embodiments, a method for diagnosing a type of amyloid disease involves administering an amyloid-reactive peptide conjugated to a detectable label. The conjugation used herein may be covalent and / or non-covalent. In some embodiments, the detectable label is covalently or non-covalently bonded to the peptide. These may include, but are not limited to, radionuclides (e.g., C-11, I-125, I-123, I-131, Zr-89, Tc-99m, Cu-64, Br-76, F-18); enzymes (horseradish peroxidase); biotin; fluorophores, etc. Any means known in the art for labeling proteins in a detectable manner may be used with and / or adapted for use with the methods described herein. For example, the amyloid-reactive peptide may be radioactively labeled with a radioisotope, or labeled with a fluorescent or chemiluminescent tag. Exemplary radioisotopes include, for example, 11 C, 18 F, 111 In, 99m Tc, and 123 I, 124 I and 125I is included. These and other radioactive isotopes can be incorporated into amyloid-reactive materials or detection dyes. Exemplary fluorescent or chemiluminescent tags include fluorescein, Texas Red, rhodamine, Alexa dye, and luciferase, which can be incorporated into amyloid-reactive materials using methods commonly used in the art.

[0098] In some embodiments, a method for diagnosing a type of amyloid disease includes administering an amyloid-reactive substance containing a radiolabel. In some embodiments, the radiolabel is 11 C, 18 F, 111 In, 99m Tc, 89 Zr and 123 I, 124 I or 125 I. In some embodiments, the radioactively labeled amyloid-reactive substance is a radioactively labeled amyloid-reactive peptide. In some embodiments, the radioactively labeled amyloid-reactive peptide is 124 It is an I-labeled amyloid-reactive peptide. In other embodiments, a method for diagnosing the type of amyloid disease is 124 This includes administering I-p5+14. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO: 13. In some embodiments, the amyloid-reactive peptide is 124 It is conjugated to I. In some embodiments, a method for diagnosing the type of amyloid disease is 124 The procedure involves administering an amyloid-reactive peptide containing SEQ ID NO: 13, conjugated to I.

[0099] In another embodiment, a method for diagnosing a type of amyloid disease includes administering an amyloid-reactive substance containing a fluorescent label. In some embodiments, the fluorescently labeled amyloid-reactive substance is thioflavin T (ThT).

[0100] In some embodiments, the amyloid-reactive material comprises an amyloid-reactive peptide conjugated to a radioactive label. In some embodiments, the amyloid-reactive material comprises a peptide conjugated to a filler. In some embodiments, the amyloid-reactive peptide is conjugated to PEG. In some embodiments, the amyloid-reactive peptide is conjugated to an antibody.

[0101] In some embodiments, amyloid-reactive substances specifically bind to amyloid deposits. In some embodiments, amyloid-reactive substances can detect the presence, absence, or amount of amyloid in a subject. In some embodiments, amyloid-reactive substances or dyes cross-react to amyloid deposits formed by a number of different proteins. In some embodiments, amyloid-reactive substances bind to amyloid deposits formed by a wide variety of proteins and / or peptides. In some embodiments, amyloid-reactive substances bind to amyloid deposits formed by amyloid light chains (AL). In some embodiments, amyloid-reactive substances bind to amyloid formed by transthyretin (TTR) fibers. In some embodiments, amyloid-reactive substances bind to amyloid formed by serum amyloid protein A (sAA). In some embodiments, the amyloid-reactive substance is immunoglobulin light chain (AL), immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin variant (ATTRv), wild-type transthyretin (ATTRwt), apolipoprotein AI (AApoAI), apolipoprotein AII variant (AApoAII), apolipoprotein AIII variant (AApoAIII), gelzolin (AGel), lysozyme (ALys), leukocyte chemotactic factor (AL It binds to the amyloid-forming forms of ECT2), fibrinogen α variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP); α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), or other amyloid-forming peptides such as IAAP, ALκ4, and ALλ1.

[0102] In some embodiments, the amyloid-reactive substance binds to heparan sulfate glycoaminoglycan (GAG). In some embodiments, the GAG ​​binds to the amyloid deposit. The binding of GAG to amyloid fibrils occurs primarily through electrostatic interactions involving negative polyvalent electrolyte charges and positively charged side chain residues of the aggregated protein. Similar to a catalyst for the reaction, GAG promotes aggregation, nucleation, and amyloid fibril formation, functioning as a structural template for the self-assembly of β-sheet-rich, highly cytotoxic oligomeric precursors into amyloid fibrils. Furthermore, the amyloid-promoting activity of GAG may be enhanced by specific interactions mediated by consensus binding sites between amyloid polypeptides and GAG molecules.

[0103] In some embodiments, the method includes administering an amyloid-reactive substance to an individual. In some embodiments, the amyloid-reactive substance is administered in a pharmaceutical composition. In some embodiments, the composition includes an aqueous buffer. The composition may also include a local anesthetic, such as lidocaine, for pain relief at the injection site, and a solubilizer. The components are supplied either mixed together in a unit dosage form, such as a lyophilized powder or anhydrous concentrate in a sealed container, such as an ampoule indicating the amount of the active substance, or individually. When the composition is administered by infusion, it may be dispensed in an infusion bottle containing sterile pharmaceutical-grade water or saline. When the composition is administered by injection, an ampoule of sterile water or saline for injection may be provided so that the components can be mixed before administration.

[0104] The composition may further comprise a carrier. The present invention also provides pharmaceutical compositions comprising one or more peptides and / or fusion peptides of the present invention. Such pharmaceutical compositions comprise an effective amount of a peptide or fusion peptide for binding to amyloid and detection of amyloid, and a pharmaceutically acceptable carrier.

[0105] Pharmaceutically acceptable carriers include solid or liquid carriers or components that may be added to enhance or stabilize the composition, or to facilitate the preparation of the composition, and these include, but are not limited to, syrups, water, isotonic saline solutions, 5% dextrose aqueous solutions or buffered sodium acetate or ammonium acetate solutions, oils, glycerin, and alcohols. Examples of oils include those of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, and sesame oil. Carriers may also contain sustained-release substances, such as glyceryl monostearate or glyceryl distearate, alone or in combination with waxes. Other suitable pharmaceutical carriers include, but are not limited to, starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, propylene glycol, water, ethanol, flavorings, preservatives, colorants, diluents, granulators, lubricants, and binders.

[0106] When pharmaceutical compositions are administered intravenously, water may be a preferred carrier. Saline solutions and aqueous dextrose and glycerol solutions may also be used as liquid carriers, particularly for injectable solutions. Compositions may optionally contain trace amounts of wetting agents or emulsifiers, or pH buffers. Such compositions may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, or sustained-release formulations. Compositions may be formulated as suppositories with common binders and carriers, such as triglycerides. Oral formulations may contain standard carriers, such as pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, or magnesium carbonate. Examples of other suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin.

[0107] Methods for imaging amyloid include, but are not limited to, magnetic resonance imaging (MRI), computed tomography (CAT) scans, positron emission tomography (PET), ultrasound imaging, X-ray imaging, radionuclide imaging, single-photon emission computed tomography (SPECT), and multiphoton microscopy.

[0108] Various contrast agents can be used to increase the sensitivity of the scan. Contrast agents for scanning may contain all molecules that attenuate X-rays. Radioisotopes can be used in positron emission tomography and radionuclide imaging. All positron emission isotopes are useful for positron emission tomography and radionuclide imaging, and all gamma-photon emission isotopes are useful for radionuclide imaging.

[0109] Contrast agents for ultrasound imaging include positive and negative agents. Positive agents reflect ultrasound energy to produce a positive (bright) image. Correspondingly, negative agents enhance transmittance or sound transmission to produce a negative (dark) image. Various substances (gases, liquids, solids, and combinations thereof) are being studied as potential contrast enhancers. Examples of solid particle contrast agents disclosed in U.S. Patent No. 5,558,854 include, but are not limited to, IDE particles and SHU454. European Patent Application 0231091 discloses an oil-in-water emulsion containing a highly fluorinated organic compound for obtaining enhanced contrast (contrast) in ultrasound imaging. Emulsions containing perfluorooctyl bromide (PFOB) are also being studied as ultrasound imaging agents. U.S. Patent No. 4,900,540 describes the use of phospholipid-based liposomes containing gases or gaseous precursors as contrast enhancers.

[0110] Several classes of compounds have potential as MRI contrast agents. These classes include superparamagnetic iron oxide particles, nitroxides, and paramagnetic metal chelates (Mann et al., 1995). Strongly paramagnetic metals are preferred. Paramagnetic lanthanides and transition metal ions are typically toxic in vivo. Therefore, these compounds need to be incorporated into chelates with organic ligands. The peptides and fusion peptides of the present invention can be used to enhance the targeting of such chelated metals to amyloid, which allows for a reduction in the total dose of imaging composition required if they were not used.

[0111] Imaging agents can be conjugated to peptides and fusion peptides using known methods. Certain conjugation methods involve the use of metal chelate complexes with organic chelating agents such as DTPA. Acceptable chelates are known in the art. These include, but are not limited to, 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA); 1,4,7,10-tetraazacyclododecane-N,N',N''-triacetic acid (DO3A); 1,4,7-tris(carboxymethyl)-10-(2-hydroxypropyl)-1,4,7,10-tetraazacyclododecane (HP-DO3A); diethylenetriaminepentaacetic acid (DTPA); and many others.

[0112] Several classes of compounds have potential as MRI contrast agents. These classes include superparamagnetic iron oxide particles, nitroxides, and paramagnetic metal chelates (Mann et al., 1995). Strongly paramagnetic metals are preferred. Paramagnetic lanthanides and transition metal ions are typically toxic in vivo. Therefore, these compounds need to be incorporated into chelates with organic ligands. The peptides and fusion peptides of the present invention can be used to enhance the targeting of such chelated metals to amyloid, which allows for a reduction in the total dose of imaging composition required if they were not used.

[0113] A wide range of paramagnetic metals are suitable for chelation. Suitable metals include those with atomic numbers 22–29 (inclusive), 42, 44, and 58–70 (inclusive) and oxidation states 2 or 3. Examples of such metals include, but are not limited to, chromium(III), manganese(II), iron(II), cobalt(II), nickel(II), copper(II), praseodymium(III), neodymium(III), samarium(III), gadolinium(III), terbium(III), dysprosium(III), holmium(III), erbium(III), ytterbium(III), and vanadium(II). Ions useful in other situations such as X-ray imaging include, but are not limited to, lanthanum(III), gold(III), lead(II), and especially bismuth(III).

[0114] Suitable radioisotopes for labeling the peptides and fusion peptides of the present invention, suitable for localization studies, include gamma emitters, positron emitters, X-ray emitters, and fluorescence emitters. Suitable radioisotopes for labeling peptides and fusion proteins include astatine. 211 ,bromine 76 , 14 carbon, 11 carbon, 51 chromium, 36 chlorine, 57 cobalt, 58 Cobalt, copper 67 ,copper 64 , 152 Europium, fluorine 18 ,gallium 67 ,gallium 68 , 3 Hydrogen, iodine 123 iodine 124 iodine 125 iodine 126 iodine 131 ,indium 111 ,indium 113m , 59 iron, 177 Lutetium, mercury 107 ,mercury 203 , 32Phosphorus, rhenium 186 , rhenium 188 , ruthenium 95 , ruthenium 97 , ruthenium 103 , ruthenium 105 , rhenium 99m , rhenium 105 , rhenium 101 , 75 selenium, 35 sulfur, technetium 99m , tellurium-121m, tellurium 122m , tellurium 125m , thulium 165 , thulium 167 , thulium 168 and yttrium 90 are included. Halogens can be used almost interchangeably as labels. Iodine, which is a gamma emitter 123 and technetium 99m can also be used. This is because such radioactive metals are detectable with a gamma camera and have advantageous half-lives for in vivo imaging. 18-fluorine, which is a positron emitter suitable for PET imaging and has a half-life suitable for peptide imaging, or 124 iodine can also be used. The peptides and fusion peptides of the present invention can be labeled with indium, via a conjugated metal chelating agent such as DTPA (diethylenetriaminepentaacetic acid), or non-covalently and directly to adjacent peptides containing Cys residues or triglycine motifs 111 or technetium 99m .

[0115] Radiolabeled peptides or fusion peptides can be produced according to methods well known in the art. For example, they can be iodized by contact with sodium iodide or potassium iodide and a chemical oxidizing agent such as sodium hypochlorite, or an enzymatic oxidizing agent such as lactoperoxidase. The peptides or fusion peptides according to the present invention can be labeled by a ligand exchange process, for example by reducing pertechnetium salt with a tin solution, chelating the reduced technetium onto a Sephadex column, and applying the peptide to this column, or by direct labeling techniques, for example by incubating pertechnetium salt, a reducing agent such as SnCl2, a buffer solution such as sodium potassium phthalate solution, and the peptide. 99m They can be labeled with . The intermediate functional groups often used to bind radioactive isotopes that exist as metal ions to peptides are, as mentioned above, diethylenetriaminepentaacetic acid (DTPA) and ethylenediaminetetraacetic acid (EDTA).

[0116] Other useful labels include fluorescent labels, chromogenic labels, and biotin labels. Fluorescent labels include, but are not limited to, rhodamine, fluorescein isothiocyanate, sodium fluorescein, lenographin, and tex-red sulfonyl chloride. In certain embodiments, the peptides and fusion peptides of the present invention may be linked to a secondary ligand or an enzyme (enzyme tag) that produces a colored product upon contact with a chromogenic substrate. Examples of suitable enzymes include urease, alkaline phosphatase, (horseradish) hydrogen peroxidase, and glucose oxidase. Secondary ligands include biotin and avidin or streptavidin compounds. The use of such markings is well known to those skilled in the art and is described, for example, in U.S. Patents No. 3,817,837, 3,850,752, 3,939,350, 3,996,345, 4,277,437, 4,275,149 and 4,366,241 (each incorporated herein by reference).

[0117] The present invention provides a method for detecting amyloid in a subject. The method comprises administering a pharmaceutical composition containing an effective amount of one or more peptides or fusion peptides of the present invention to a subject, and detecting the peptides or fusion peptides bound to amyloid. The peptides may be labeled with imaging agents, such as radioisotopes. The peptides have specific binding affinity to the deposits, and the binding is detectable. Binding of the peptides or fusion peptides to amyloid can be detected by MRI, CAT scan, PET imaging, ultrasound imaging, SPECT imaging, X-ray imaging, fluorescence imaging, or radionuclide imaging.

[0118] In some examples, a method for diagnosing a type of amyloid disease involves administering a detectable amount of amyloid-reactive substance or dye to an individual. The detectable amount to be administered may depend on the type of detection being performed. For example, in some embodiments, the detectable amount of amyloid-reactive substance or dye may be sufficient to be detectable by imaging when administered to the subject. The detectable amount of amyloid-reactive substance to be administered to an individual may vary depending on factors such as the individual's age, sex and weight, the individual's specific response, dose measurement, formulation, and equipment-related factors. Optimization of such factors is well within the scope of the art. The detectable amount of amyloid-reactive substance may also vary depending on the form of administration of the amyloid-reactive substance or detection dye.

[0119] In some cases, amyloid-reactive substances are administered parenterally, paracancerally, transmucosally, percutaneously, intramuscularly, intravenously, intradermally, subcutaneously, intraperitoneally, intraventricularly, or intracranially. In some cases, amyloid-reactive substances are administered intravenously. In other cases, amyloid-reactive substances are administered intraperitoneally.

[0120] Those skilled in the art will further understand that an effective amount of amyloid-reactive material may be administered in a single dose (single administration) or may be achieved by multiple doses (multiple administrations). In some cases, the administration of the amyloid-reactive material may further include the administration of a flushing solution. For example, a flushing solution, such as saline, may be administered immediately after the administration of the amyloid-reactive material or detection dye, or after a certain period following the administration of the amyloid-reactive material or detection dye. In other cases, the amyloid-reactive material may be metabolized and excreted within a certain period after administration.

[0121] C. Detection of amyloid in organs and tissues In some embodiments, a method for diagnosing amyloid-related disease includes detecting amyloid with an amyloid-reactive substance or a detection dye. Examples of amyloids that may be detected as part of this method include, but are not limited to, the amyloid-forming forms of immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin variants (ATTR), amyloid beta (Aβ), apolipoprotein AI (AApoAI), apolipoprotein AII (AApoAII), gelzolin (AGel), lysozyme (ALys), leukocyte chemotactic factor (ALect2), fibrinogen α variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP); α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), or IAAP, and other amyloid-forming peptides. In some embodiments of this disclosure, a method for diagnosing a type of amyloid disease includes detecting ATTR, AL, and / or ALECT2 amyloid. In other embodiments, a method for diagnosing a type of amyloid disease includes distinguishing between ATTR, AL, and ALECT2 amyloid.

[0122] In some embodiments, the method may be for diagnosing amyloid-related diseases. In some embodiments, amyloid-related diseases are AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, Alys, ALECT2, AFib, ACys, ACa, AMed, AIAPP, APro, AIns, APrP, or Aβ amyloidosis. In some embodiments, amyloid-related diseases are systemic amyloidosis. In some embodiments, amyloid-related diseases are cardiac amyloidosis. In some embodiments, amyloid-related diseases are cardiac AL amyloidosis. In some embodiments, amyloid-related diseases are cardiac ATTR amyloidosis.

[0123] In some embodiments, a method for diagnosing amyloid-related disease comprises administering an amyloid-reactive peptide conjugated to a detectable label to an individual, and measuring the amount of the detectable label in the tissue or organ of the individual, where an amount of signal from the amyloid-reactive peptide exceeding a threshold indicates that the individual has amyloid disease.

[0124] In some embodiments, the method includes administering to an individual a dose of about 0.1 mCi to about 25 mCi of amyloid-reactive peptide conjugated to a detectable label. In some embodiments, the method includes administering to an individual a dose of about 0.3 mCi, 1 mCi, or about 2 mCi of amyloid-reactive peptide conjugated to a detectable label. In some embodiments, the method includes administering to an individual a dose of 1 mCi (±10%) of amyloid-reactive peptide conjugated to a detectable label. In some embodiments, the method includes administering to an individual a dose of about 0.9 mCi to about 1.1 mCi of amyloid-reactive peptide conjugated to a detectable label. In some embodiments, the method includes administering to an individual a dose of 1 mCi (±10%) and 2 mCi or less of amyloid-reactive peptide conjugated to a detectable label. In some embodiments, an amyloid-reactive peptide conjugated to a detectable label is 124 It is an I-labeled amyloid-reactive peptide.

[0125] In some embodiments, the method is 124 The method involves administering a dose of approximately 0.1 mCi to approximately 25 mCi of I-labeled amyloid-reactive peptide to an individual. In some embodiments, the method is as follows: 124 The method involves administering a dose of approximately 0.3 mCi, 1 mCi, or approximately 2 mCi of I-labeled amyloid-reactive peptide to an individual. In some embodiments, the method is as follows: 124 The method comprises administering a dose of 1 mCi (±10%) of I-labeled amyloid-reactive peptide to an individual. In some embodiments, the method is 124 The method involves administering a dose of approximately 0.9 mCi to approximately 1.1 mCi of I-labeled amyloid-reactive peptide to an individual. In some embodiments, the method is as follows: 124 This includes administering to an individual a dose of 1 mCi (±10%) and 2 mCi or less of I-labeled amyloid-reactive peptide.

[0126] In some embodiments, the method involves administering an individual about 0.3 mg, about 1 mg, about 1.5 mg, or about 2 mg of amyloid-reactive peptide. In some embodiments, the method involves administering an individual about 0.3 mg to about 2 mg, about 1 mg to about 2 mg, or about 1.5 mg to about 2 mg of amyloid-reactive peptide. In some embodiments, the method involves administering an individual about 0.3 mg to about 1 mg, or about 0.3 mg to about 1.5 mg of amyloid-reactive peptide.

[0127] In some embodiments, the step of detecting amyloid-reactive material in one or more organs, as in the method, includes determining organ uptake values ​​for each organ. Organ uptake can be determined by methods known to those skilled in the art. For example, organ uptake values ​​may represent relative or absolute levels of amyloid-reactive material detected in each organ of an individual. In some embodiments, the organ uptake value ratio is the relative uptake value. In some embodiments, the organ uptake value is the standard uptake value (SUV) for each organ. In some embodiments, as will be understood to those skilled in the art, the SUV may be determined by measuring the amount of amyloid-reactive material detected in a reactive organ (e.g., heart) compared to the amount of amyloid-reactive material detected in a non-reactive tissue or sample (e.g., blood). The amount of amyloid-reactive material in an organ may be determined, for example, by quantifying the detectable signal from the amyloid-reactive material in the organ (e.g., by calculating the pixel value in an image). In some embodiments, the standard uptake value is determined as the ratio of the amount of amyloid-reactive substance detected in an organ to the amount of amyloid-reactive substance detected in the blood. In some embodiments, the organ uptake value indicates the organ distribution pattern of the amyloid-reactive substance or detection dye. In some embodiments, the method includes determining the organ-specific SUV for an individual. In some embodiments, the organ-specific SUV for an individual is selected from the group consisting of SUV mean, SUV max, and SUV peak.

[0128] In some embodiments, organ uptake is based on organ-specific standard uptake ratios (SUVRs). In some embodiments, SUVR is calculated using a blood pool as a reference tissue. In some embodiments, SUVR is calculated for each organ by dividing the amount of amyloid-detecting substance or dye in the organ by the blood pool ratio. In some embodiments, the blood pool is a vein or artery. In some embodiments, the blood pool is the lumen of the thoracic aorta. In some embodiments, the organ-specific SUVR for an individual is selected from the group consisting of SUVR mean, SUVR max, and SUVR peak. In some embodiments, the organ-specific SUVR for an individual is the SUVR mean. In some embodiments, organ-specific SUVR is used to adjust for (control) differences in the clearance rates of radioactive tracers.

[0129] In some embodiments, the threshold is the SUVR mean threshold. In some embodiments, the SUVR mean threshold is an organ- or tissue-based SUVR mean threshold. The SUVR mean threshold may be selected according to the organ or tissue in which amyloid-reactive peptides conjugated to a detectable label are detected. In some embodiments, the organ or tissue is the liver, spleen, kidney, pancreas, heart, or adrenal gland. In some embodiments, the detection of amyloid-reactive peptides conjugated to a detectable label above the SUVR mean threshold level enables the diagnosis of early-stage amyloid-related disease.

[0130] In some embodiments, the threshold is the mean SUVR threshold. In some embodiments, if the organ or tissue is the liver, the mean SUVR threshold is 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, or 1.6. In some embodiments, if the organ or tissue is the liver, the mean SUVR threshold is 1.31. In some embodiments, if the mean SUVR in an individual's liver exceeds the threshold of 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, or 1.6, the individual is likely to have systemic amyloidosis or a certain type of systemic amyloidosis. In some embodiments, if the SUVR value in an individual's liver exceeds the threshold of 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, or 1.6, the individual is diagnosed with systemic amyloidosis or a certain type of systemic amyloidosis.

[0131] In some embodiments, the threshold is the mean SUVR threshold. In some embodiments, if the organ or tissue is the spleen, the mean SUVR threshold is 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5. In some embodiments, if the organ or tissue is the spleen, the mean SUVR threshold is 1.21. In some embodiments, if the mean SUVR in an individual's spleen exceeds the threshold of 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5, the individual is likely to have systemic amyloidosis or a certain type of systemic amyloidosis. In some embodiments, if the mean SUVR in an individual's spleen exceeds the threshold of 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5, the individual is diagnosed with systemic amyloidosis or a certain type of systemic amyloidosis.

[0132] In some embodiments, the threshold is the mean SUVR threshold. In some embodiments, if the organ or tissue is the kidney, the mean SUVR threshold is 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, or 2.3. In some embodiments, if the organ or tissue is the kidney, the mean SUVR threshold is 1.88 (or between 1.69 and 2.07). In some embodiments, if the mean SUVR in an individual's kidney exceeds the threshold of 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, or 2.3, the individual is likely to have systemic amyloidosis or a certain type of systemic amyloidosis. In some embodiments, an individual is diagnosed with systemic amyloidosis or a certain type of systemic amyloidosis if the mean SUVR in the individual's kidney exceeds a threshold of 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, or 2.3.

[0133] In some embodiments, the threshold is the mean SUVR threshold. In some embodiments, if the organ or tissue is the pancreas, the mean SUVR threshold is 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, or 1.7. In some embodiments, if the organ or tissue is the pancreas, the mean SUVR threshold is 1.40. In some embodiments, if the mean SUVR in an individual's pancreas exceeds the threshold of 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, or 1.7, the individual is likely to have systemic amyloidosis or a certain type of systemic amyloidosis. In some embodiments, if the mean SUVR in an individual's pancreas exceeds the threshold of 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, or 1.7, the individual is diagnosed with systemic amyloidosis or a certain type of systemic amyloidosis.

[0134] In some embodiments, the threshold is the mean SUVR threshold. In some embodiments, if the organ or tissue is the heart, the mean SUVR threshold is 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, or 2.1. In some embodiments, if the organ or tissue is the heart, the mean SUVR threshold is 1.19. In some embodiments, if the mean SUVR in an individual's heart exceeds the threshold of 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, or 2.1, the individual is likely to have systemic amyloidosis or a certain type of systemic amyloidosis. In some embodiments, an individual is diagnosed with systemic amyloidosis or a certain type of systemic amyloidosis if the mean SUVR in the individual's heart exceeds a threshold of 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, or 2.1.

[0135] In some embodiments, the threshold is the mean SUVR threshold. In some embodiments, if the organ or tissue is the adrenal gland, the mean SUVR threshold is 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, or 1.4. In some embodiments, if the organ or tissue is the adrenal gland, the mean SUVR threshold is 1.135. In some embodiments, if the mean SUVR in an individual's adrenal gland exceeds the threshold of 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, or 1.4, the individual is likely to have systemic amyloidosis or a certain type of systemic amyloidosis. In some embodiments, if the mean SUVR in an individual's adrenal gland exceeds the threshold of 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, or 1.4, the individual is diagnosed with systemic amyloidosis or a certain type of systemic amyloidosis.

[0136] In some embodiments, the threshold is the SUVR maximum threshold. In some embodiments, the SUVR maximum threshold is an organ or tissue-based SUVR maximum threshold. The SUVR maximum threshold may be selected according to the organ or tissue in which amyloid-reactive peptides conjugated to a detectable label are detected. In some embodiments, the organ or tissue is the liver, spleen, kidney, pancreas, heart, or adrenal gland. In some embodiments, the detection of amyloid-reactive peptides conjugated to a detectable label above the SUVR maximum threshold level enables the diagnosis of early-stage amyloid-related disease.

[0137] In some embodiments, the threshold is the SUVR maximum threshold. In some embodiments, when the organ or tissue is the liver, the SUVR maximum threshold is 10.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12. The values ​​are 9, 13, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15, 15.1, 15.2, 15.3, 15.4, 15.5, 15.6, 15.7, 15.8, 15.9, or 16. In some embodiments, when the organ or tissue is the liver, the maximum SUVR threshold is 13.26.

[0138] In some embodiments, the threshold is the SUVR maximum threshold. In some embodiments, when the organ or tissue is the spleen, the SUVR maximum threshold is 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, or 5.9. In some embodiments, when the organ or tissue is the spleen, the SUVR maximum threshold is 4.86.

[0139] In some embodiments, the threshold is a SUVR maximum threshold. In some embodiments, when the organ or tissue is the kidney, the SUVR maximum threshold is 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6 or 11.7. In some embodiments, when the organ or tissue is the kidney, the SUVR maximum threshold is 9.68.

[0140] In some embodiments, the threshold is a SUVR maximum threshold. In some embodiments, when the organ or tissue is the pancreas, the SUVR maximum threshold is 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3 or 11.4. In some embodiments, when the organ or tissue is the pancreas, the SUVR maximum threshold is 9.48.

[0141] In some embodiments, the threshold is a SUVR maximum threshold. In some embodiments, when the organ or tissue is the heart, the SUVR maximum threshold is 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4 or 7.5. In some embodiments, when the organ or tissue is the heart, the SUVR maximum threshold is 6.2.

[0142] In some embodiments, the threshold is a SUVR maximum threshold. In some embodiments, when the organ or tissue is the adrenal gland, the SUVR maximum threshold is 1, 1.1, 1.2, 1.3, 1.4, 1.5 or 1.6. In some embodiments, when the organ or tissue is the adrenal gland, the SUVR maximum threshold is 1.31.

[0143] In some embodiments, the step of measuring the organ uptake value or the organ uptake value ratio of the amyloid-reactive substance in one or more organs comprises analyzing imaging data. The imaging data may be generated by any procedure known in the art that can enable imaging of the amyloid-reactive substance or dye. For example, the amyloid-reactive substance may be detected by positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI) or single-photon emission computed tomography (SPECT). In certain embodiments, the amyloid-reactive substance may be detected by a combined imaging method, for example PET / CT (PET performed simultaneously with computed tomography imaging) or PET / MRI (PET performed simultaneously with magnetic resonance imaging). The imaging method may provide one or more images of the observation region of the subject. In certain embodiments, imaging yields a plurality of images, which may be combined, superimposed, added, subtracted, color-coded, or otherwise fused and mathematically manipulated by any method known in the art. The generated image may be a digital or analog image that may be displayed, for example, as a "hard" image on printer paper, photographic paper or film, or as an image on a screen such as a video or LCD screen.

[0144] In some embodiments, the PET image is analyzed using the region of interest (ROI) method. In some embodiments, the image is a planar image. In some embodiments, the image is a coronal, axial or sagittal image.

[0145] In some embodiments, the threshold for diagnosing a particular type of amyloid disease is selected based on the desired sensitivity. In some embodiments, the cutoff value is selected so as to yield a sensitivity of at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%. In some embodiments, the sensitivity for detecting cardiac amyloid using an amyloid-reactive peptide conjugated to a detectable label is about 80% to about 100%. In some embodiments, the sensitivity for detecting cardiac amyloid using an amyloid-reactive peptide conjugated to a detectable label is about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100%. In some embodiments, the sensitivity for detecting cardiac amyloid using an amyloid-reactive peptide conjugated to a detectable label is about 96%.

[0146] In some embodiments, the sensitivity for detecting cardiac amyloid in patients with AL amyloidosis is approximately 80% to approximately 100%. In some embodiments, the sensitivity for detecting cardiac amyloid in patients with AL amyloidosis is approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, approximately 99%, or approximately 100%. In some embodiments, the sensitivity for detecting cardiac amyloid in patients with AL amyloidosis is approximately 93%.

[0147] In some embodiments, the sensitivity for detecting cardiac amyloid in patients with ATTR amyloidosis is approximately 80% to approximately 100%. In some embodiments, the sensitivity for detecting cardiac amyloid in patients with ATTR amyloidosis is approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, approximately 99%, or approximately 100%. In some embodiments, the sensitivity for detecting cardiac amyloid in patients with ATTR amyloidosis is approximately 100%.

[0148] In some embodiments, the threshold is selected based on the desired specificity (i.e., the ability to exclude individuals who do not have amyloid-related disease). In some embodiments, the cutoff value is selected so as to yield a specificity of at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%.

[0149] In some embodiments, the threshold is selected based on organ-specific SUV or SUVR values ​​to obtain the optimal relationship between sensitivity and specificity. In some embodiments, the threshold is selected to obtain the highest Youden's index. In some embodiments, the Youden's index is determined by the following formula: Sensitivity + Specificity - 1. In some embodiments, if several SUVs or SUVRs give the same Youden's index, the SUV or SUVR that yields the highest specificity is selected.

[0150] In some embodiments, the average cardiac SUVR for AL amyloidosis (SUVR) mean The threshold is approximately 0.6 to 4. In some embodiments, the mean cardiac SUVR threshold for AL amyloidosis is approximately 1.0, 1.1, 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, 3, 3.5, or 4.0.

[0151] In some embodiments, the mean cardiac SUVR threshold for ATTR amyloidosis is approximately 0.6 to approximately 4. In some embodiments, the mean cardiac SUVR threshold for ATTR amyloidosis is approximately 1.0, approximately 1.1, approximately 1.2, approximately 1.4, approximately 1.6, approximately 1.8, approximately 2.0, approximately 2.2, approximately 2.4, approximately 2.6, approximately 2.8, approximately 3, approximately 3.5, or approximately 4.0.

[0152] In some embodiments, the sensitivity and specificity of amyloid-reactive peptides conjugated to a detectable label are defined in comparison to clinically approved methods for diagnosing amyloid-related diseases. In some embodiments, clinically approved methods are selected from the group consisting of an individual's health-related quality of life indicators, the presence or amount of one or more biomarkers associated with amyloid-related diseases, cardiac biopsy, and additional amyloid imaging of the individual. In some embodiments, the sensitivity and specificity of amyloid-reactive peptides conjugated to a detectable label are defined in comparison to diagnoses obtained from the individual's medical records.

[0153] In some embodiments, the sensitivity of an amyloid-reactive peptide is the specificity of the amyloid-reactive peptide conjugated to a detectable label. In some embodiments, the sensitivity of an amyloid-reactive peptide conjugated to a detectable label is measured by dividing the number of true positives by the number of false negatives. In some embodiments, the number of true positives is the number of individuals who actually have amyloid-related disease and are correctly diagnosed with amyloid-related disease by the amyloid-reactive peptide conjugated to a detectable label. In some embodiments, the number of false negatives is the number of individuals who actually have amyloid-related disease and are incorrectly diagnosed as not having amyloid-related disease by the amyloid-reactive peptide conjugated to a detectable label. In some embodiments, sensitivity may be defined as a confidence interval of the sensitivity value. In some embodiments, sensitivity may be defined according to a 95% confidence interval of the sensitivity value. In some embodiments, the 95% confidence interval may be calculated using the Wilson-Brown method.

[0154] In some embodiments, the sensitivity of an amyloid-reactive peptide conjugated to a detectable label is the sensitivity of detecting amyloid-related disease after administration of the amyloid-reactive peptide and detection of the radioactive label by amyloid imaging. In some embodiments, the sensitivity of an amyloid-reactive peptide conjugated to a detectable label is related to the dose of the amyloid-reactive peptide conjugated to a detectable label administered to an individual.

[0155] In some embodiments, the specificity of the amyloid-reactive peptide is the specificity of the amyloid-reactive peptide conjugated to a detectable label. In some embodiments, the specificity of the amyloid-reactive peptide conjugated to a detectable label is measured by dividing the number of true negatives by the sum of the number of false positives. In some embodiments, the number of true negatives is the number of individuals who were correctly diagnosed as not having amyloid-related disease by the amyloid-reactive peptide conjugated to a detectable label, when the individuals did not actually have amyloid-related disease. In some embodiments, the number of false positives is the number of individuals who were diagnosed with amyloid-related disease by the amyloid-reactive peptide conjugated to a detectable label, when the individuals did not actually have amyloid-related disease. In some embodiments, specificity may be defined as a confidence interval of the sensitivity value. In some embodiments, specificity may be defined according to a 95% confidence interval of the specificity value. In some embodiments, the 95% confidence interval may be calculated using the Wilson-Brown method.

[0156] In some embodiments, the specificity of an amyloid-reactive peptide conjugated to a detectable label is the specificity of detecting amyloid-related disease after administration of the amyloid-reactive peptide and detection of the radioactive label by amyloid imaging. In some embodiments, the specificity of an amyloid-reactive peptide conjugated to a detectable label is related to the dose of the amyloid-reactive peptide conjugated to a detectable label administered to the individual. In some embodiments, the step of detecting amyloid-reactive material is performed in multiple organs in the individual. In some embodiments, the detection step further includes measuring the organ distribution pattern of the amyloid-reactive material or detection dye, where the organ distribution pattern of the amyloid-reactive material indicates the type of amyloid disease. The type of amyloid disease may be sporadic amyloidosis or may have a genetic component (e.g., hereditary amyloidosis). Non-exclusive examples of amyloid diseases include AA amyloidosis, AL amyloidosis, AH amyloidosis, Aβ amyloidosis, ATTR amyloidosis, ALect2 amyloidosis, as well as IAPP amyloidosis in type 2 diabetes, Alzheimer's disease, Down syndrome, hereditary cerebral hemorrhage with Dutch amyloidosis, cerebral beta-amyloid angiopathy, cavernous encephalopathy, thyroid tumors, Parkinson's disease, Lewy body dementia, tauopathy, Huntington's disease, senile systemic amyloidosis, familial hemodialysis, senile systemic aging, senile pituitary disorders, iatrogenic syndromes, cavernous encephalopathy, reactive chronic inflammation, thyroid tumors, myeloma or other forms of cancer. In some embodiments, the type of amyloid disease is systemic amyloid disease. In some embodiments, the type of amyloid disease is AL amyloidosis, ATTR amyloidosis, or ALECT2 amyloidosis. In some embodiments, the amyloid disease is cardiac amyloidosis. In some embodiments, the amyloid disease is cardiac AL amyloidosis. In some embodiments, the amyloid disease is cardiac ATTR amyloidosis.

[0157] In some embodiments, a method for diagnosing amyloid-related disease includes measuring the organ distribution pattern of amyloid-reactive material in one or more organs. While not bound by theory, it is believed that the anatomical distribution of amyloid in each form of the disease may have a specific pattern. For example, in ATTR amyloidosis, amyloid deposition is common in the heart and peripheral nerves, while another common amyloidosis, AL amyloidosis, exhibits diverse amyloid deposition patterns, with amyloid being observed, for example, in the heart, spleen, liver, kidneys, peripheral nerves, gastrointestinal tract, muscles, lungs, and lymph nodes. In some embodiments, a method for diagnosing the type of amyloid disease includes measuring the organ distribution pattern of amyloid-reactive material in one or more of the following: heart, spleen, liver, kidneys, peripheral nerves, gastrointestinal tract, muscles, lungs, brain, and lymph nodes. In some embodiments, the one or more organs are abdominal and thoracic organs. In some embodiments, the one or more organs are the heart, spleen, liver, or kidneys.

[0158] In other examples, a method for diagnosing amyloid-related disease includes administering an amyloid-reactive substance and calculating an inter-organ ratio for two or more organs. In some examples, the step of calculating an inter-organ ratio for two or more organs includes calculating the ratio of organ uptake for a first organ to organ uptake for a second organ. In some examples, the inter-organ ratio is selected from the group consisting of liver vs. heart, spleen vs. heart, spleen vs. liver, spleen vs. kidney, kidney vs. heart, and kidney vs. liver. In some examples, the inter-organ ratio is the heart vs. spleen ratio. In some examples, the inter-organ ratio is between 0 and 1, 1, or greater than 1. In some examples, the inter-organ ratio indicates the type of amyloid disease in an individual.

[0159] In some embodiments, organ uptake values ​​or inter-organ ratios indicate the type of amyloid disease. In some embodiments, organ uptake values ​​or inter-organ ratios indicate the type of amyloid disease only if they exceed a cutoff value or threshold. For example, in some embodiments, if the inter-organ ratio for a certain type of amyloid disease is 1.4, a diagnosis of that type of amyloid disease is made if the inter-organ ratio calculated for an individual is 1.4 or greater. As another non-limiting example, if the cutoff for organ uptake values ​​for a certain type of amyloid disease is 1.4, a diagnosis of that type of amyloid disease is inappropriate if the inter-organ ratio calculated for an individual is less than 1.4. Specific cutoff values ​​or thresholds for diagnosing the type of amyloid disease may vary depending on the type of amyloid disease, the stage of disease progression, the patient's statistical attributes, the administered amyloid-reactive substance, and the detection method used. In some embodiments, organ uptake values ​​or inter-organ cutoff values ​​or thresholds are calculated from data on the organ distribution of the amyloid-reactive substance or detection dye. In some embodiments, organ uptake values ​​or inter-organ cutoff values ​​or thresholds are calculated from data from a population with a specific type of amyloid disease.

[0160] Preferably, the cutoff value or threshold for organ uptake values ​​or inter-organ uptake ratios for diagnosing the type of amyloid disease is determined using a receiver operating characteristic curve. As understood in the art, a receiver operating characteristic curve (ROC curve) plots the performance of a particular feature for distinguishing between two groups: patients with amyloid disease and controls (e.g., those without amyloid disease). Data across the entire population (i.e., patients and controls) are sorted in ascending order based on the value of a single feature (e.g., organ uptake value). Then, for each value of that feature, the true positive rate and false positive rate for the data are determined. The true positive rate (sensitivity) is determined by counting the number of cases that exceed the value of the feature under consideration and then dividing by the total number of patients. The false positive rate (specificity) is determined by counting the number of controls that exceed the value of the feature under consideration and then dividing by the total number of controls.

[0161] ROC curves can be created not only for a single feature but also for other single outputs. For example, a combination of two or more features can be mathematically summed (addition, plotting, multiplication, etc.) to obtain a single sum, which can then be plotted on the ROC curve. Furthermore, any combination of multiple features that results in a single output value can be plotted on the ROC curve. These combinations of features can be included in a test. The ROC curve plots the true positive rate (sensitivity) of a test against the false positive rate (1-specificity) of the test. The area under the ROC curve (AUC) can be a performance index for a given sample population, ranging from 0 for a test that shows a completely random response when classifying test subjects to 1 for a perfect test. As with any diagnostic application, the area under the ROC curve indicates the predictive power of a model and can be used to compare the predictive power of one model with another. Using ROC curves, cutoff values ​​can be selected to diagnose an individual's amyloid disease and / or amyloid type with high reliability.

[0162] In some embodiments, the method includes obtaining organ distribution data relating to amyloid-reactive substances or detection dyes. In some embodiments, the organ distribution data is an image. Images generated using the imaging methods embodied in the present invention can be analyzed by any method known in the art. For example, in some embodiments, imaging data obtained from a PET or SPECT scan can be input to a processor that identifies individual pixels or groups of pixels whose brightness is greater than a predetermined threshold or mean background value, and the identified pixels can be characterized as indicating the presence of amyloid-reactive substances or dyes. In another embodiment, image data can be obtained from an image scanned and input to a processor. In such embodiments, it is possible to identify amyloid-reactive substances or dyes in the image using a similar process that identifies bright areas on the image. In certain embodiments, the analysis of the image may further include determining the intensity, density, strength or combination thereof of the output brightness, which may correlate with the amount of radioactively labeled protein in the image, a region or area of ​​the image or a specific location on the image. While we do not wish to be bound by theory, areas or locations on an image that have a higher intensity than other areas or locations may hold a higher concentration of, for example, radiolabeled proteins that target amyloid deposition, and therefore may have a higher concentration of radiolabeled amyloid-reactive substances or dyes attached to the areas where amyloid-reactive substances or dyes are localized.

[0163] In some embodiments, a method for diagnosing the type of amyloid disease comprises analyzing an image by the spatial location of a region of interest targeted by an administered amyloid-reactive substance or a detectable dye. In other embodiments, analysis of the pharmacokinetics of the administered amyloid-reactive substance or dye can provide information regarding the appropriate injection timing of the amyloid-reactive substance or dye. The presence or absence of amyloid can be determined by identifying a region, range, or location on the image that correlates with the presence of the radiolabeled protein. For example, in some embodiments, identification of a region or location where the amyloid-reactive substance is concentrated indicates the presence of amyloid. In some embodiments, an image that correlates with the presence of the amyloid-reactive substance is used to diagnose amyloid disease in an individual.

[0164] In some embodiments, the method further comprises providing a diagnosis of the type of amyloid disease based on an organ distribution pattern. In some embodiments, a specific organ distribution pattern indicates a specific type of amyloid disease. For example, in some embodiments, the ratio of heart to spleen, heart to liver, spleen to liver, spleen to kidney, kidney to heart, or kidney to liver is used for the diagnosis of ATTR. In some embodiments, the ratio of heart to spleen, heart to liver, spleen to liver, spleen to kidney, kidney to heart, or kidney to liver is used for the diagnosis of ALECT2. In some embodiments, the ratio of heart to spleen, heart to liver, spleen to liver, spleen to kidney, kidney to heart, or kidney to liver is used for the diagnosis of AL.

[0165] In some embodiments, different types of amyloid diseases have different relative inter-organ ratios. In some embodiments, a certain type of amyloid disease can have a higher liver-to-heart ratio than another type. In some embodiments, the heart-to-spleen ratio for an individual diagnosed with ATTR is higher than the heart-to-spleen ratio for an individual diagnosed with AL. In some embodiments, the heart-to-spleen ratio for an individual diagnosed with ATTR is higher than the heart-to-spleen ratio for an individual diagnosed with ALECT2.

[0166] In some embodiments, the level of amyloid-reactive material in the heart is highest in individuals with ATTR. In some embodiments, the level of amyloid-reactive material in the liver is highest in individuals with AL. In some embodiments, the level of amyloid-reactive material is highest in the spleen of individuals with ALECT2. In some embodiments, the level of amyloid-reactive material is highest in the kidneys of individuals with ALECT2.

[0167] In some embodiments, the level of amyloid-reactive material is lowest in the heart of individuals with ALECT2. In some embodiments, the level of amyloid-reactive material is lowest in the liver of individuals with ATTR. In some embodiments, the level of amyloid-reactive material is lowest in the spleen of individuals with ATTR. In some embodiments, the level of amyloid-reactive material is lowest in the kidneys of individuals with ATTR.

[0168] In some embodiments, the cutoff value for diagnosing a particular type of amyloid disease is selected based on a specific p-value. In some embodiments, the cutoff value is selected to provide p-values ​​less than 0.1, less than 0.05, less than 0.01, less than 0.005, or less than 0.001.

[0169] Those skilled in the art will understand that each ratio considered herein can be easily converted to its reciprocal. For example, a heart-to-spleen ratio of 2:1 (2) is the same as a spleen-to-heart ratio of 1:2 (0.5).

[0170] In some embodiments, the diagnosis is confirmed by a measure of the individual's health-related quality of life. In some embodiments, the diagnosis is confirmed by the presence or amount of one or more biomarkers associated with amyloid-related disease. In some embodiments, the diagnosis is confirmed by cardiac biopsy. In some embodiments, the diagnosis is confirmed by additional amyloid imaging of the individual.

[0171] In some embodiments, the method further includes administering potassium iodide to the individual. Potassium iodide can protect the individual's thyroid gland. In some embodiments, potassium iodide is administered orally. In some embodiments, 130 mg of potassium iodide is administered for 7 days, starting one day before administration of the amyloid-reactive substance. In some embodiments, 130 mg of potassium iodide is administered for 3 days, starting on the day of administration of the amyloid-reactive substance, for example, about 30 minutes before administration of the amyloid-reactive substance.

[0172] D. Patient population In some embodiments, the method for diagnosing the type of amyloid disease is particularly useful in individuals at risk of developing amyloid-related disease. In some embodiments, individuals at risk of developing amyloid-related disease have a family history of amyloid-related disease. In some embodiments, individuals are at risk of developing hereditary amyloid-related disease. In some embodiments, the genetic form of amyloid-related disease includes hereditary transthyretin amyloidosis (hATTR).

[0173] In some embodiments, an individual is determined to be at risk of amyloid-related disease based on the presence of a gene mutation. In some embodiments, the gene mutation is an ATTR gene mutation. In some embodiments, the gene mutation is located within the TTR gene. In some embodiments, the gene mutation includes a point mutation resulting in an amino acid substitution, such as a substitution of valine to methionine at position 30 (Val30Met).

[0174] In some embodiments, an individual is determined to be at risk of amyloid-related disease based on having multiple myeloma. In some embodiments, an individual at risk of developing amyloid-related disease has smoldering multiple myeloma. In some embodiments, an individual is at risk of developing immunoglobulin light chain amyloidosis (AL) based on having multiple myeloma.

[0175] In some embodiments, an individual is determined to be at risk of amyloid-related disease based on having unspecified monoclonal gammaglobulinemia (MGUS).

[0176] In some embodiments, an individual is determined to be at risk of amyloid-related disease based on having amyloid-positive laminectomy tissue. In some embodiments, an individual is determined to be at risk of amyloid-related disease based on having amyloid-positive carpal tunnel release tissue. In some embodiments, an individual is determined to be at risk of amyloid-related disease based on being elderly. In some embodiments, the individual is at least about 60, 65, 70, 75, 80, 85, or 90 years old. In some embodiments, the individual is at least 80 years old.

[0177] In some embodiments, individuals at risk of developing amyloid-related disease exhibit symptoms of amyloid-related disease. In some embodiments, the symptoms of amyloid-related disease are selected from the group consisting of swelling of the ankles and legs, severe fatigue, shortness of breath, significant weight loss, difficulty swallowing, tingling, numbness or pain in the hands, wrists or feet, enlargement of the tongue, irregular heartbeat, diarrhea, and easy bruising, thickening or purple spots on the skin.

[0178] Alternatively, in some embodiments, individuals at risk of developing amyloid-related disease do not exhibit symptoms of amyloid-related disease.

[0179] In some embodiments, the individual is suspected to have amyloid-related disease. In some embodiments, the individual is suspected to have amyloid-related disease but has not been diagnosed with amyloid-related disease. In some embodiments, the individual has not undergone a diagnostic evaluation for amyloid-related disease. In some embodiments, the individual has undergone a diagnostic evaluation for amyloid-related disease, but the evaluation did not detect amyloid-related disease. In some embodiments, the individual has not been diagnosed according to the Mayo Clinic System. In some embodiments, the individual suspected to have amyloid-related disease exhibits symptoms of amyloid-related disease. Alternatively, in some embodiments, the individual is at risk of developing amyloid-related disease but does not exhibit symptoms of amyloid-related disease. In some embodiments, the individual suspected to have amyloid-related disease has a genetic predisposition to amyloid-related disease. In some embodiments, the individual suspected to have amyloid-related disease has a family history of some form of amyloid-related disease. In some embodiments, individuals suspected of having amyloid-related disease are elderly. In some embodiments, individuals are in the early stages of amyloid-related disease.

[0180] E. Prognosis Furthermore, this specification provides a method for determining the prognosis of an individual having an amyloid-related disease, the method comprising: a) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label to the individual, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs to quantify the amyloid in the individual at a first time point; b) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label to the individual, and detecting the amount of the detectable label in the individual's tissues or organs to quantify the amyloid in the individual at a second time point; and c) determining the individual's prognosis by comparing the amyloid quantified at the first time point in the tissues or organs with the amyloid quantified at the second time point in the tissues or organs.

[0181] In some embodiments, the individual has an amyloid disorder selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, Alys, ALECT2, AFib, ACys, ACa, AMed, AIAPP, APro, AIns, APrP, or Aβ amyloidosis. In some embodiments, the individual has an early-stage amyloid-related disease. In some embodiments, the individual has stage 1 amyloidosis. In some embodiments, the individual has systemic amyloidosis. In some embodiments, the individual has cardiac amyloidosis. In some embodiments, the individual has cardiac AL amyloidosis. In some embodiments, the individual has cardiac ATTR amyloidosis.

[0182] In some embodiments, the interval between a first time point in time in which the amyloid-reactive peptide conjugated to a detectable label is administered and a second time point in time in which the amyloid-reactive peptide conjugated to a detectable label is administered is at least 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, or 5 years. In some embodiments, the first and second time points are separated by at least 2 weeks. In some embodiments, the first and second time points are separated by at least 1 month. In some embodiments, the first and second time points are separated by at least 3 months.

[0183] In some embodiments, the prognosis of individuals with amyloid-related disease is worsened. In some embodiments, the prognosis of individuals with amyloid-related disease is improved.

[0184] In some embodiments, prognosis is confirmed by indicators of quality of life related to the individual's health. In some embodiments, prognosis is confirmed by the presence or amount of one or more biomarkers associated with amyloid-related disease. In some embodiments, prognosis is confirmed by cardiac biopsy. In some embodiments, prognosis is confirmed by additional amyloid imaging of the individual.

[0185] III. Treatment method Furthermore, this specification provides a method for treating amyloid-related diseases, the method comprising: a) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label to an individual; b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs; and c) performing treatment for amyloid-related diseases if the amount of the amyloid-reactive peptide exceeds a threshold.

[0186] This specification provides a method for treating amyloid-related disease in an individual having an early-stage amyloid-related disease, the method comprising administering an amyloid-reactive peptide conjugated to a detectable label to the individual, detecting the amount of the detectable label in the individual's tissues or organs, and, if the amount of the amyloid-reactive peptide exceeds a threshold, performing treatment for the amyloid-related disease. In some embodiments, an amount of amyloid-reactive peptide exceeding a threshold indicates that the individual has amyloid-related disease, where the amyloid-related disease is an early-stage amyloid-related disease. In some embodiments, the amyloid-related disease is systemic amyloidosis. In some embodiments, the amyloid-related disease is cardiac amyloidosis. In some embodiments, the amyloid-reactive peptide conjugated to a detectable label is 124 It is a peptide containing the amino acids listed in SEQ ID NO: 100, conjugated to I. In some embodiments, 124 I is bonded to amino acid 4 of sequence number 100.

[0187] The methods provided herein may be used to treat early-stage amyloid-related diseases. In some embodiments, the methods may be used to treat amyloid-related diseases before the disease becomes detectable by conventional methods. In some embodiments, the individual has been previously diagnosed as not having amyloid-related disease. In some embodiments, the methods have high sensitivity and specificity for detecting and treating early-stage amyloid-related diseases.

[0188] A. Administration of amyloid-reactive peptides Some aspects of the present invention provide methods for treating amyloid diseases based on detecting amyloid using an amyloid-reactive substance or a detection dye. In some embodiments, methods for treating amyloid diseases are provided herein, which include administering an amyloid-reactive substance or a detection dye, measuring the organ distribution of the amyloid-reactive substance or detection dye, and selecting a treatment based on the type of disease.

[0189] In some embodiments, a method for treating amyloid disease includes administering an amyloid-reactive substance or a detection dye, and measuring organ-specific SUV and / or SUVR of the amyloid-reactive substance with respect to one or more organs. In some embodiments, the organ distribution pattern of the amyloid-reactive substance or dye indicates the type of amyloid disease. In some embodiments, the method further includes selecting a treatment based on the type of amyloid disease.

[0190] In some embodiments, the method includes obtaining an organ distribution pattern of an amyloid-reactive substance or detection dye (where the organ distribution pattern indicates a particular type of amyloid disease), and carrying out treatment based on the amyloid disease.

[0191] In some embodiments, a method for treating amyloid disease involves administering an amyloid-reactive substance comprising an amyloid-reactive peptide conjugated to a detectable label. In some embodiments, the amyloid-reactive substance comprises a peptide, a fusion protein, a small molecule compound, or an antibody or fragment.

[0192] In some embodiments, a method for treating amyloid disease includes administering an amyloid-reactive substance comprising an amyloid-reactive peptide conjugated to a detectable label. In some embodiments, the amyloid-reactive peptide is a peptide having the amino acid sequence described in any one of SEQ ID NOs: 1 to 14. In some embodiments, a method for treating amyloid disease includes an amyloid-reactive peptide having the amino acid sequence described in SEQ ID NO: 13.

[0193] In some embodiments, a method for treating amyloid disease includes administering an amyloid-reactive substance or a detection dye, and measuring organ-specific SUV and / or SUVR of the amyloid-reactive substance with respect to one or more organs. In some embodiments, a method for treating amyloid disease includes administering an amyloid-reactive substance containing a detectable label to determine the organ distribution pattern. In some embodiments, the amyloid-reactive substance includes a fluorescent label, a chemiluminescent tag, or a radioactive label. In some embodiments, the amyloid-reactive substance includes a radioactive label. In some embodiments, the radioactively labeled amyloid-reactive substance is 124 In some embodiments, the radioactively labeled amyloid-reactive substance is I-p5+14. In other embodiments, the radioactively labeled amyloid-reactive substance is flolbetapyr, floltaben, or flutemetamol. In some embodiments, the treatment method for amyloid disease includes p5+14. In some embodiments, the amyloid-reactive substance is radioactively labeled. In some embodiments, the radioactive label is 11 C, 18 F, 111 In, 99m Tc and 123 I, 124 I or 125 I. In some embodiments, the radioactively labeled amyloid-reactive substance is a radioactively labeled amyloid-reactive peptide. In some embodiments, the radioactively labeled amyloid-reactive peptide is 124It is an I-labeled amyloid-reactive peptide. In other embodiments, a method for diagnosing the type of amyloid disease is 124 This includes administering I-p5+14. In other embodiments, the radiolabeled amyloid-reactive substance is flolbetapyr, floltaben, or flutemetamol.

[0194] In certain other embodiments, the amyloid-reactive substance includes a fluorescent label. In some embodiments, the fluorescently labeled amyloid-reactive substance is ThT. In some embodiments, the amyloid-reactive substance is administered parenterally, paracancerally, transmucosally, percutaneously, intramuscularly, intravenously, intradermally, subcutaneously, intraperitoneally, intraventricularly, or intracranially. In some examples, the amyloid-reactive substance is administered intravenously or intraperitoneally.

[0195] In some embodiments, the method further includes administering potassium iodide to the individual. Potassium iodide can protect the individual's thyroid gland. In some embodiments, potassium iodide is administered orally. In some embodiments, 130 mg of potassium iodide is administered for 7 days, starting one day before administration of the amyloid-reactive substance. In some embodiments, 130 mg of potassium iodide is administered for 3 days, starting on the day of administration of the amyloid-reactive substance, for example, about 30 minutes before administration of the amyloid-reactive substance.

[0196] B. Detection of amyloid levels In some embodiments, a method for treating amyloid disease includes administering an amyloid-reactive substance or a detection dye and measuring organ-specific SUV and / or SUVR of the amyloid-reactive substance in one or more organs. In some embodiments, a method for treating amyloid disease includes measuring the organ distribution pattern of the amyloid-reactive substance in one or more organs. In some embodiments, a method for treating this type of amyloid disease includes measuring organ-specific SUV and / or SUVR of the amyloid-reactive substance in one or more of the following: lungs, fat, heart, kidneys, pancreas, joints, spine, liver, spleen, adrenal glands, bone lesions, choroid plexus, pituitary gland, uterus, bone marrow, musculoskeletal tissue, gastrointestinal tract, and prostate. In some embodiments, the one or more organs are the heart, spleen, liver, or kidneys. In some embodiments, the step of measuring the organ distribution pattern of the amyloid-reactive substance in one or more organs includes determining organ uptake values ​​for each organ. In some embodiments, the organ uptake values ​​are standard uptake values ​​for each organ. In some embodiments, the standard uptake value is determined as the ratio of the amount of amyloid-reactive substance detected in an organ to the amount of amyloid-reactive substance detected in the blood. In some embodiments, the organ uptake value indicates the organ distribution pattern of the amyloid-reactive substance or detection dye.

[0197] In some embodiments, measuring organ-specific SUV and / or SUVR of amyloid-reactive material involves analyzing imaging data generated by PET, CT, MRI, SPECT, PET / CT, PET / MRI, or other imaging techniques. In some embodiments, the step of measuring organ-specific SUV and / or SUVR of amyloid-reactive material involves analyzing images based on the spatial location of the region of interest.

[0198] In other embodiments, the step of measuring the organ distribution pattern of amyloid-reactive material in one or more organs includes calculating an inter-organ ratio for two or more organs. In some embodiments, the step of calculating an inter-organ ratio for two or more organs includes calculating the ratio of organ uptake values ​​for a first organ to organ uptake values ​​for a second organ. In some examples, the inter-organ ratio is selected from the group consisting of liver vs. heart, spleen vs. heart, spleen vs. liver, spleen vs. kidney, kidney vs. heart, and kidney vs. liver. In some examples, the inter-organ ratio is the heart vs. spleen ratio. In some embodiments, the ratio is the reciprocal of any of these ratios.

[0199] In some embodiments, the organ distribution pattern indicates the type of amyloid disease. In some embodiments, the organ distribution pattern is used to select a specific treatment based on the type of amyloid disease. In some embodiments, the method further includes providing a diagnosis of the type of amyloid disease based on the organ distribution pattern. In some embodiments, a specific organ distribution pattern indicates a specific type of amyloid disease. For example, in some embodiments, the ratio of heart to spleen, heart to liver, spleen to liver, spleen to kidney, kidney to heart, or kidney to liver is used to diagnose ATTR. In some embodiments, the ratio of heart to spleen, heart to liver, spleen to liver, spleen to kidney, kidney to heart, or kidney to liver is used to diagnose ALECT2. In some embodiments, the ratio of heart to spleen, heart to liver, spleen to liver, spleen to kidney, kidney to heart, or kidney to liver is used to diagnose AL.

[0200] C. Implementation of treatment In some embodiments, the method includes treating an amyloid-related disease in an individual, or selecting a treatment for an amyloid-related disease in an individual. Some examples of amyloid diseases that may be diagnosed and / or treated by the methods disclosed herein include, but are not limited to, AA amyloidosis, AL amyloidosis, AH amyloidosis, Aβ amyloidosis, ATTR amyloidosis, ALECT2 amyloidosis, and IAPP amyloidosis in type 2 diabetes, Alzheimer's disease, thyroid tumors, Parkinson's disease, tauopathy, senile systemic amyloidosis, familial hemodialysis, senile systemic aging, senile pituitary disorders, iatrogenic syndromes, cavernous encephalopathy, reactive chronic inflammation, thyroid tumors, myeloma or other forms of cancer. In some embodiments, the method for treating an amyloid disease includes selecting a treatment for systemic amyloidosis. In some embodiments, a method for treating amyloid disease includes selecting a treatment for AL amyloidosis, ATTR amyloidosis, or ALECT2 amyloidosis. In some embodiments, the treatment is targeted therapy for ATTR amyloidosis, AL amyloidosis, or ALECT2 amyloidosis. In some embodiments, a method for treating amyloid disease includes selecting a treatment for cardiac amyloidosis. In some embodiments, a method for treating amyloid disease includes selecting a treatment for cardiac AL amyloidosis. In some embodiments, a method for treating amyloid disease includes selecting a treatment for cardiac ATTR amyloidosis. In some embodiments, the amyloid-related disease is in its early stages. In some embodiments, the amyloid-related disease is an early-stage amyloid-related disease.

[0201] In some embodiments, the method includes treating amyloid-related disease or selecting a treatment for amyloid-related disease in an individual suspected of having amyloid-related disease. In some embodiments, the individual is suspected of having amyloid-related disease but has not been diagnosed with amyloid-related disease. In some embodiments, the individual has not undergone a diagnostic evaluation for amyloid-related disease. In some embodiments, the individual has undergone a diagnostic evaluation for amyloid-related disease, but the diagnostic evaluation did not detect amyloid-related disease. In some embodiments, the individual has not been diagnosed according to the Mayo Clinic System. In some embodiments, the individual suspected of having amyloid-related disease exhibits symptoms of amyloid-related disease. Alternatively, in some embodiments, an individual at risk of developing amyloid-related disease does not exhibit symptoms of amyloid-related disease. In some embodiments, the individual suspected of having amyloid-related disease has a genetic predisposition to amyloid-related disease. In some embodiments, individuals suspected of having amyloid-related disease have a family history of any amyloid-related disease. In some embodiments, individuals suspected of having amyloid-related disease are elderly. In some embodiments, individuals are in the early stages of amyloid-related disease.

[0202] In some embodiments, the treatment is a small molecule, antibody, peptide, protein, nucleic acid, and / or gene therapy. In some embodiments, the treatment is a targeted therapy specific to a particular type of amyloid disease.

[0203] In some embodiments, the treatment is an antibody-peptide fusion comprising an antibody-peptide fusion protein containing a second amyloid-reactive peptide and an antibody that binds to amyloid fibrils. In some embodiments, the second amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 14, including 0, 1, 2, 3, or 4 amino acid substitutions, insertions, or deletions. In some embodiments, the second amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises (i) an amyloid-reactive peptide comprising the amino acid sequence described in SEQ ID NO: 2, and (ii) an antibody that binds to human amyloid fibrils.

[0204] In some embodiments, the antibody that binds to human amyloid fibrils includes a heavy chain and a light chain. In some embodiments, the antibody that binds to human amyloid fibrils includes a heavy chain variable region (VH), and the light chain of the antibody includes a light chain variable region (VL) according to Table 2. In some embodiments, VH contains the amino acids listed in SEQ ID NO: 102. In some embodiments, VL contains the amino acids listed in SEQ ID NO: 101. [Table 2]

[0205] In some embodiments, the heavy chain of the antibody includes a heavy chain variable region (VH), and the light chain of the antibody includes a light chain variable region (VL), where VH and VL include CDRs according to Table 3. In some embodiments, the heavy chain of the antibody includes a heavy chain variable region (VH), and the light chain of the antibody includes a light chain variable region (VL), where VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 106, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 107, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 108, and VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 103, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 104, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 105. [Table 3]

[0206] In some embodiments, the amyloid-reactive peptide and the antibody are linked at the C-terminus of the light chain, where the amyloid-reactive peptide is linked to the antibody via a spacer containing the amino acid sequence: VSPSV (SEQ ID NO: 83). In some embodiments, the antibody-peptide fusion may be one of the modified immunoglobulins described in U.S. Patent Application Publication No. 12,030,934B2.

[0207] In some embodiments, the treatment is a targeted therapy for ATTR amyloidosis, AL amyloidosis, or ALECT2 amyloidosis. In some embodiments, the treatment is a targeted therapy for ATTR amyloidosis. In some embodiments, the treatment comprises a TTR tetramer stabilizer. In some embodiments, the TTR tetramer stabilizer is epigallocatechin-3-gallate (EGCG), AG-10, CHF5074, tafadimis, or diflunisal. In some embodiments, the treatment comprises an antibody or fragment that binds to misfolded TTR. In some embodiments, the antibody is PRX004. In some embodiments, the treatment comprises an oligonucleotide. In some embodiments, the oligonucleotide is a TTR silencer. In some embodiments, the TTR silencer is patisirane (ALN-TTR02), butrisilane, inotercene, or AKCEA-TTR-LRx. In some embodiments, the treatment comprises an ATTR amyloid-destroying agent. In some embodiments, the treatment comprises doxycycline, taurourusdeoxycholic acid, or serum amyloid-P (SAP). In some embodiments, the treatment comprises organ transplantation. In some embodiments, the treatment comprises liver transplantation.

[0208] In other embodiments, the treatment is targeted therapy for AL amyloidosis. In some embodiments, the treatment comprises bortezomib, ixazomib, or kaffirzomib. In some embodiments, the treatment comprises an antibody or fragment. In some embodiments, the treatment comprises daratumumab, CAEL-101, elotuzumab, or verantamab mafodotin. In some embodiments, the treatment comprises stem cell therapy. In some embodiments, the treatment comprises a corticosteroid. In some embodiments, the corticosteroid is dexamethasone.

[0209] In some embodiments, the method is used to rule out potential therapies for patients with amyloidosis. In some embodiments, the method is used to diagnose one type of amyloidosis and to rule out therapies for other types of amyloidosis. In some embodiments, the method is used to diagnose ALECT2 and to rule out therapies for AL or ATTR amyloidosis.

[0210] In some embodiments, the method is used to differentiate types of amyloidosis in order to develop therapies specific to a particular type of amyloidosis. For example, in some embodiments, the method is used to identify individuals with ALECT2 amyloidosis and to develop therapies specific to ALECT2 amyloidosis.

[0211] In some embodiments, the present invention provides a method for monitoring the treatment of amyloid-related disease in an individual, the method comprising: a) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label to the individual and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs to quantify amyloid in the individual at a first time point; b) administering treatment for amyloid-related disease; c) administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label to the individual and detecting the amount of the detectable label in the individual's tissues or organs to quantify amyloid in the individual at a second time point; and d) comparing the amyloid quantified in the tissues or organs at the first time point with the amyloid quantified in the tissues or organs at the second time point to determine whether the treatment is effective.

[0212] In some embodiments, the interval between a first time point in time when the amyloid-reactive peptide is administered and a second time point in time when the amyloid-reactive peptide is administered is at least 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, or 5 years. In some embodiments, the first and second time points are separated by at least 2 weeks. In some embodiments, the first and second time points are separated by at least 1 month. In some embodiments, the first and second time points are separated by at least 3 months.

[0213] In some embodiments, this specification provides a method for selecting a treatment for amyloid-related disease in an individual, the method comprising: a) administering to the individual an amyloid-reactive peptide comprising an amino acid sequence described in any one of SEQ ID NOs: 1 to 14 conjugated to a detectable label; and b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the tissue or organ of the individual, wherein if amyloid is detected in the heart, a treatment for amyloid-related disease is performed, and if amyloid is not detected in the heart, an alternative therapy is performed.

[0214] IV. Kit Some aspects of the present invention provide kits for diagnosing or detecting types of amyloid disease in an individual using the methods described herein.

[0215] In some embodiments, the kit includes an amyloid-reactive substance and instructions for use. In some embodiments, the amyloid-reactive substance includes a detectable label. In some embodiments, the amyloid-reactive substance is 124 It is I-p5+14. In some embodiments, the amyloid-reactive material includes an amino acid sequence containing SEQ ID NO: 13.

[0216] In some embodiments, the radioactive label is 11 C,18 F, 111 In, 99m Tc, 123 I, 124 I or 125 I. In some embodiments, the radioactively labeled amyloid-reactive substance is a radioactively labeled amyloid-reactive peptide. In some embodiments, the radioactively labeled amyloid-reactive peptide is 124 It is an I-labeled amyloid-reactive peptide. In other embodiments, a method for diagnosing the type of amyloid disease is 124 This includes administering I-p5+14. In other embodiments, the radiolabeled amyloid-reactive substance is flolbetapyr, floltaben, or flutemetamol.

[0217] In some embodiments, the instructions for use include instructions for detecting amyloid-reactive material in one or more organs. In some embodiments, the amyloid-reactive material is detected in the blood, heart, lungs, kidneys, or spleen.

[0218] In some embodiments, the kit includes instructions for calculating organ-specific SUV. In some embodiments, the kit includes instructions for calculating organ-specific SUVR. In some embodiments, the kit includes instructions for calculating SUVR ratios for one or more organs. In some embodiments, SUVR is calculated using a blood pool as the reference tissue. In some embodiments, the SUVR for each organ is calculated by dividing the amount of amyloid-detecting substance or dye in the organ by the blood pool ratio. In some embodiments, the blood pool is a vein or artery. In some embodiments, the blood pool is the lumen of the thoracic aorta.

[0219] In some embodiments, the instructions for use include instructions for determining inter-organ ratios. In some embodiments, the inter-organ ratios are liver to heart, spleen to heart, spleen to liver, spleen to kidney, kidney to heart, kidney to liver, or the reciprocal of any of these ratios.

[0220] In some embodiments, the kit further includes instructions for providing a diagnosis based on organ-specific SUV or SUVR. In some embodiments, the kit includes instructions for providing a diagnosis based on inter-organ ratios.

[0221] In some embodiments, the kit includes a therapeutic substance for treating amyloid disease.

[0222] Exemplary Embodiments 1. A method for diagnosing amyloid-related disease in an individual at risk of developing amyloid-related disease, wherein the method is: a. Administering an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs. 1-14, conjugated to a detectable label, to an individual, and b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual. The method comprising, wherein the amount of amyloid-reactive peptide exceeding a threshold indicates that the individual has an amyloid disease.

[0223] 2. The method according to Embodiment 1 for determining that an individual is at risk of amyloid-related disease based on the presence of a gene mutation, having multiple myeloma, having amyloid-positive laminectomy tissue, having amyloid-positive tissue derived from carpal tunnel release surgery, having monoclonal immunoglobulinemia of unknown significance (MGUS), having heart failure with preserved ejection fraction (HFpEF), having heart failure with reduced ejection fraction (HFrEF), being from a susceptible ethnic group, or being elderly.

[0224] 3. The method according to Embodiment 2, wherein the gene mutation is located within the transthyretin protein.

[0225] 4. The method according to Embodiment 2, wherein the gene mutation is located within the fibrinogen α protein.

[0226] 5. The method according to any one of Embodiments 1 to 3, wherein the individual does not exhibit symptoms of amyloidosis.

[0227] 6. The method according to any one of Embodiments 1 to 3, wherein the individual has neurological symptoms of amyloid disease.

[0228] 7. A method for diagnosing amyloid-related disease in an individual suspected of having amyloid-related disease, the method being: a. Administering an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs. 1-14, conjugated to a detectable label, to an individual, and b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual. The method comprising, wherein the amount of amyloid-reactive peptide exceeding a threshold indicates that the individual has an amyloid disease.

[0229] 8.a. Quantify amyloid in an individual by administering an amyloid-reactive peptide containing the amino acid sequence described in one of SEQ ID NOs: 1-14, conjugated to a detectable label, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs. b. Administer an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs: 1-14, conjugated to a detectable label, to an individual, and quantify amyloid in the individual at a second time point by detecting the amount of the detectable label in the individual's tissues or organs, and c. Determining the prognosis of an individual by comparing the amyloid quantified at a first time point in the tissue or organ with the amyloid quantified at a second time point in the same tissue or organ. A method for determining the prognosis of individuals with amyloid-related diseases, including [specific example].

[0230] 9. The method according to Embodiment 8, wherein the prognosis of an individual with amyloid-related disease is based on detecting the amount of amyloid-reactive peptide in the heart only.

[0231] 10. The method according to Embodiment 8, wherein the prognosis of an individual with amyloid-related disease is based on detecting the amount of amyloid-reactive peptide in the heart and kidneys.

[0232] 11. The method according to Embodiment 8, wherein the prognosis of an individual with amyloid-related disease is based on detecting the amount of amyloid-reactive peptide in the heart, kidneys and all other organs.

[0233] 12.a. Administering an individual an amyloid-reactive peptide containing the amino acid sequence described in one of SEQ ID NOs: 1-14, conjugated to a detectable label. b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual, and c. When the amount of amyloid-reactive peptide exceeds a threshold, treatment for amyloid-related disease should be initiated. Methods for treating amyloid-related diseases, including [specific example].

[0234] 13.a. Quantifying amyloid in an individual by administering an amyloid-reactive peptide containing the amino acid sequence described in one of SEQ ID NOs: 1-14, conjugated to a detectable label, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs. b. To provide treatment for amyloid-related diseases. c. Administer an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs: 1-14, conjugated to a detectable label, to an individual, and quantify amyloid in the individual at a second time point by detecting the amount of the detectable label in the individual's tissues or organs, and d. To determine whether the treatment is effective by comparing the amyloid quantified at the first time point in the tissue or organ with the amyloid quantified at the second time point in the same tissue or organ. A method for monitoring the treatment of amyloid-related diseases in an individual, including [specific example].

[0235] 14. The method according to any one of Embodiments 1 to 13, further comprising administering an amyloid-reactive peptide containing an amino acid sequence described in any one of Sequence IDs 1 to 14 conjugated to a detectable label to an individual, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs, thereby quantifying amyloid in the individual at a second time point, optionally a third time point, a fourth time point, and / or a fifth time point.

[0236] 15. The method according to embodiment 13 or 14, wherein the first time point and the second time point are separated by at least six weeks.

[0237] 16. A method for selecting a treatment for amyloid-related disease in an individual, the method being: a. Administering an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs. 1-14, conjugated to a detectable label, to an individual, and b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual. The method comprising, wherein if amyloid is detected in the heart, treatment for amyloid-related disease is performed, and if amyloid is not detected in the heart, alternative therapy is performed.

[0238] 17.a. Quantifying amyloid in an individual by administering an amyloid-reactive peptide containing the amino acid sequence described in one of SEQ ID NOs: 1-14, conjugated to a detectable label, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs. b. To provide treatment for amyloid-related diseases. c. Administer an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs: 1-14, conjugated to a detectable label, to an individual, and quantify amyloid in the individual at a second time point by detecting the amount of the detectable label in the individual's tissues or organs, and d. Comparing amyloid quantified at a first time point in a tissue or organ with amyloid quantified at a second time point in the same tissue or organ. e. Adjusting treatment for amyloid-related diseases. A method for managing the treatment of amyloid-related diseases in an individual, including the treatment of amyloid-related diseases in an individual.

[0239] 18. The method according to Embodiment 17, wherein the treatment is modified if the amyloid disease is stable between the first and second time points.

[0240] 19. The method according to Embodiment 18, wherein treatment is resumed if the amyloid load is increased.

[0241] 20. The method according to any one of Embodiments 1 to 19, wherein the amyloid-reactive peptide comprises SEQ ID NO: 13.

[0242] 21. The method according to any one of Embodiments 1 to 20, wherein the detectable label is a radioactive label.

[0243] 22. Detectable signs are 11 C, 18 F, 111 In, 99m Tc, 89 Zr, 68 Ga and 123 I,124 I and 125 A method according to any one of Embodiments 1 to 21, selected from the group consisting of I.

[0244] 23. Detectable signs 124 The method described in any one of Embodiments 1 to 22, which is I.

[0245] 24. Doses of approximately 0.1 to 25 mCi 124 A method according to any one of Embodiments 1 to 23, comprising administering an I-labeled amyloid-reactive peptide to an individual.

[0246] 25. Doses of approximately 0.3 mCi, 1 mCi, or 2 mCi 124 A method according to any one of Embodiments 1 to 24, comprising administering an I-labeled amyloid-reactive peptide to an individual.

[0247] 26. A method according to any one of Embodiments 1 to 25, comprising administering a dose of Tc-99 of approximately 20 mCi to an individual.

[0248] 27. A method according to any one of Embodiments 1 to 26, comprising administering to an individual approximately 0.3 mg, 1 mg, 1.5 mg, or approximately 2 mg of amyloid-reactive peptide.

[0249] 28. The method according to any one of Embodiments 1 to 27, wherein an amyloid-reactive peptide is administered intravenously.

[0250] 29. The method according to any one of Embodiments 1 to 28, further comprising determining organ-specific standard uptake ratios for organs within an organism.

[0251] 30. The method according to Embodiment 29, wherein the organ-specific SUVR for an individual is selected from the group consisting of SUVR mean, SUVR maximum, and SUVR peak.

[0252] 31. The method according to any one of Embodiments 1 to 30, further comprising determining organ-specific SUVs for an individual.

[0253] 32. The method according to Embodiment 31, wherein the organ-specific SUV for an individual is selected from the group consisting of SUV mean, SUV maximum, and SUV peak.

[0254] 33. The method according to any one of Embodiments 1 to 32, further comprising determining the organ-specific dose per gram percentage (%ID) for an individual.

[0255] 34. The method according to any one of Embodiments 1 to 33, wherein the amyloid-reactive peptide has at least 90% sensitivity.

[0256] 35. The method according to any one of Embodiments 1 to 34, wherein the sensitivity is approximately 99% or approximately 100%.

[0257] 36. The method according to any one of Embodiments 1 to 35, further comprising determining one or more health-related quality of life indicators for an individual.

[0258] 37. The method according to any one of Embodiments 1 to 36, further comprising detecting one or more biomarkers associated with amyloid-related disease.

[0259] 38. The method according to Embodiment 37, wherein the biomarker associated with amyloid-related disease is selected from the group consisting of troponin T, NTproBNP, urinary protein levels, UACR, EGFR, and alkaline phosphatase levels.

[0260] 39. The method according to any one of Embodiments 1 to 38, wherein the tissue or organ of the individual is selected from the group consisting of lungs, fat, heart, kidneys, pancreas, joints, spine, liver, spleen, adrenal gland, bone lesions, choroid plexus, pituitary gland, uterus, bone marrow, musculoskeletal tissue, gastrointestinal tract, and prostate.

[0261] 40. The method according to any one of Embodiments 1 to 39, further comprising performing a cardiac biopsy if amyloid is detected in the heart.

[0262] 41. The method according to any one of Embodiments 1 to 40, further comprising performing additional amyloid imaging on an individual.

[0263] 42. The method according to Embodiment 41, wherein additional amyloid imaging of the individual includes ECHO, CMR, bone scintigraphy, or positron emission tomography imaging.

[0264] 43. Additional amyloid imaging of the individual, 99m Tc-PyP, 99m Tc-DPD, 99m Tc-HMDP, 99m Tc-MDP, other bone scintigraphy tracers, 124 I and 18 F-Florbetapyr, 18 F-Frutemtamol and 18 The method according to Embodiment 41, further comprising a tracer selected from the group consisting of F-fluorbetabene.

[0265] 44. The method according to any one of Embodiments 1 to 43, wherein the diagnosis, prognosis, or response is confirmed by an indicator of the individual's health-related quality of life, the presence or amount of one or more biomarkers associated with amyloid-related disease, cardiac biopsy, and / or additional amyloid imaging of the individual.

[0266] 45. The method according to any one of Embodiments 1 to 44, wherein an individual is diagnosed with amyloid cardiomyopathy.

[0267] 46. ​​The method according to any one of Embodiments 1 to 45, wherein the individual is suspected to have amyloid cardiomyopathy.

[0268] 47. The method according to any one of Embodiments 1 to 46, wherein the amyloid-related disease is systemic or localized amyloidosis.

[0269] 48. The method according to any one of Embodiments 1 to 47, wherein the amyloid-reactive peptide has pan-amyloid specificity.

[0270] 49. Amyloid-reactive peptides include immunoglobulin light chain (AL), immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin (ATTR wild-type; ATTR mutant), apolipoprotein AI (AApoAI), apolipoprotein AII (AApoAII), apolipoprotein AIV (AApoAIV), gelzolin (AGel), apolipoprotein C-II (AApoCII), apolipoprotein C-III (AApoCIII), lysozyme (ALys), and leukemia. A method according to any one of Embodiments 1 to 48, which binds to amyloid of a bulbar chemotactic factor (ALECT2), fibrinogen α variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP); α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), IAAP, ALβ4, or ALβ1.

[0271] 50. The method according to any one of Embodiments 1 to 49, wherein amyloid-reactive peptides are detected using PET, PET / CT, gamma machinistography, SPECT, and / or SPECT / CT.

[0272] 51. The method according to any one of Embodiments 1 to 50, wherein the amyloid-related disease is selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AApoCII, AApoCIII, AGel, Alys, ALECT2, AFib, ACys, ACa, AMed, AIAPP, APro, AIns, APrP, and Aβ amyloidosis.

[0273] 52. The method according to any one of Embodiments 1 to 51, wherein the individual has a genetic predisposition to amyloid-related disease.

[0274] 53. The method according to any one of Embodiments 1 to 52, wherein the individual has a family history of amyloid-related disease.

[0275] 54. The method according to any one of Embodiments 1 to 53, wherein the individual is elderly.

[0276] 55. The method according to any one of Embodiments 1 to 54, wherein the individual has an early-stage amyloid-related disease.

[0277] 56. The method according to any one of Embodiments 1 to 55, wherein the individual has early-stage AL amyloidosis.

[0278] 57. The method according to Embodiment 56, wherein early-stage AL amyloidosis is diagnosed according to the Mayo Clinic system.

[0279] 58. The method according to Embodiment 57, wherein the early stage AL amyloidosis is stage 1 AL amyloidosis.

[0280] 59. The method according to any one of Embodiments 1 to 58, wherein the individual has early-stage ATTR amyloidosis.

[0281] 60. The method according to Embodiment 59, wherein early-stage ATTR amyloidosis includes stage 1 ATTR amyloidosis.

[0282] 61. The method according to any one of embodiments 1-11, 14-16, and 18-60, further comprising administering a therapy for amyloid-related disease.

[0283] 62. The method according to any one of Embodiments 12-13, 17, and 61, wherein the therapy for amyloid-related disease is selected from the group consisting of transthyretin stabilizers (e.g., tafamidis, acolamidis), transthyretin silencers (e.g., patisirane, inotercene, butricirane, eprotercene), gene editing approaches, anti-amyloid approaches utilizing monoclonal antibodies, therapies targeting plasma cell clones (e.g., daratumumab, bortezomib), and antibody-peptide fusions comprising an antibody-peptide fusion protein containing a second amyloid-reactive peptide and an antibody that binds to amyloid fibril.

[0284] 63. The method according to Embodiment 62, wherein the second amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 14, comprising 0, 1, 2, 3, or 4 amino acid substitutions, insertions, or deletions.

[0285] 64. The method according to Embodiment 62 or 63, wherein the antibody-peptide fusion protein comprises a heavy chain and a light chain, and an amyloid-reactive peptide is ligated to the C-terminus of the antibody's light chain via a spacer.

[0286] 65. Antibody-peptide fusion proteins, (i) an amyloid-reactive peptide containing the amino acid sequence described in Sequence ID No. 2, and (ii) Antibodies that bind to human amyloid fibrils The method according to any one of embodiments 62 to 64, wherein the antibody comprises a heavy chain and a light chain, the heavy chain of the antibody comprises a heavy chain variable region (VH), and the light chain of the antibody comprises a light chain variable region (VL), wherein VH comprises CDR-H1 comprising the amino acid sequence described in SEQ ID NO: 106, CDR-H2 comprising the amino acid sequence described in SEQ ID NO: 107, and CDR-H3 comprising the amino acid sequence described in SEQ ID NO: 108, and VL comprises CDR-L1 comprising the amino acid sequence described in SEQ ID NO: 103, CDR-L2 comprising the amino acid sequence described in SEQ ID NO: 104, and CDR-L3 comprising the amino acid sequence described in SEQ ID NO: 105, wherein the amyloid-reactive peptide and the antibody are linked at the C-terminus of the light chain, and the amyloid-reactive peptide is linked to the antibody via a spacer comprising the amino acid sequence described in SEQ ID NO: 83.

[0287] 66. The method according to any one of Embodiments 1 to 65, wherein a peptide comprising an amyloid-reactive peptide and an N-terminal leader sequence is administered to an individual.

[0288] 67. The method according to Embodiment 66, wherein the peptide containing the amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO: 100.

[0289] 68. The method according to Embodiment 67, wherein a detectable label is bound to one or more amino acids at positions 1 to 6 of the amino acid sequence described in Sequence ID No. 100.

[0290] 69. Detectable signs 124 The method described in Embodiment 68, which is I.

[0291] 70. 124 The method according to Embodiment 69, wherein I is bound to amino acid 4 of SEQ ID NO: 100.

[0292] 71. A peptide comprising the amino acids described in Sequence ID No. 100, wherein the peptide comprises a detectable label bound to one or more amino acids at positions 1 to 6.

[0293] 72. Detectable signs 124 Peptide I according to Embodiment 71.

[0294] 73. 124 The peptide according to Embodiment 72, wherein I is bound to amino acid 4 of SEQ ID NO: 100.

[0295] 1A. A method for diagnosing early-stage amyloid-related disease in individuals at risk of developing amyloid-related disease, wherein the method is: a. Administering an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs. 1-14, conjugated to a detectable label, to an individual, and b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual. The method comprising, where the amount of amyloid-reactive peptide exceeding a threshold indicates that the individual has an amyloid-related disease, and the amyloid-related disease is an early stage of amyloid-related disease.

[0296] 2A. The method described in Embodiment 1A for determining that an individual is at risk of amyloid-related disease based on the presence of a gene mutation, having multiple myeloma, having amyloid-positive laminectomy tissue, having amyloid-positive tissue derived from carpal tunnel release surgery, having monoclonal immunoglobulinemia of unknown significance (MGUS), having heart failure with preserved ejection fraction (HFpEF), having heart failure with reduced ejection fraction (HFrEF), being from a susceptible ethnic group, or being elderly.

[0297] 3A. The method according to Embodiment 2A, wherein the gene mutation is located within the transthyretin protein.

[0298] 4A. The method according to Embodiment 2A, wherein the gene mutation is located within the fibrinogen α protein.

[0299] 5A. The method according to any one of Embodiments 1A to 3A, wherein the individual does not exhibit symptoms of amyloidosis.

[0300] 6A. The method according to any one of Embodiments 1A to 3A, wherein the individual has neurological symptoms of amyloid disease.

[0301] 7A. The method according to any one of Embodiments 1A to 6A, wherein the individual has been previously diagnosed as not having amyloid-related disease.

[0302] 8A. A method for diagnosing early-stage amyloid-related disease in an individual suspected of having amyloid-related disease, the method being: a. Administering an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs. 1-14, conjugated to a detectable label, to an individual, and b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual. The method comprising, where the amount of amyloid-reactive peptide exceeding a threshold indicates that the individual has an amyloid-related disease, and the amyloid-related disease is an early stage of amyloid-related disease.

[0303] 9A. A method for determining the prognosis of an individual with an early-stage amyloid-related disease, wherein the method is: a. Quantifying amyloid in an individual by administering an amyloid-reactive peptide containing an amino acid sequence described in one of SEQ ID NOs: 1-14, conjugated to a detectable label, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs. b. Administer an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs: 1-14, conjugated to a detectable label, to an individual, and quantify amyloid in the individual at a second time point by detecting the amount of the detectable label in the individual's tissues or organs, and c. Determining the prognosis of an individual by comparing the amyloid quantified at a first time point in the tissue or organ with the amyloid quantified at a second time point in the same tissue or organ. The method comprising, where amyloid-related disease is an early stage of amyloid-related disease.

[0304] 10A. The method according to Embodiment 9A, wherein the prognosis of an individual with amyloid-related disease is based on detecting the amount of amyloid-reactive peptide in the heart only.

[0305] 11A. The method according to Embodiment 9A, wherein the prognosis of an individual with amyloid-related disease is based on detecting the amount of amyloid-reactive peptide in the heart and kidneys.

[0306] 12A. The method according to Embodiment 9A, wherein the prognosis of an individual with amyloid-related disease is based on detecting the amount of amyloid-reactive peptide in the heart, kidneys and all other organs.

[0307] 13A.a. Administering an individual an amyloid-reactive peptide containing the amino acid sequence described in one of SEQ ID NOs: 1-14, conjugated to a detectable label. b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual, and c. When the amount of amyloid-reactive peptide exceeds a threshold, treatment for amyloid-related disease should be initiated. A method for treating amyloid-related disease in individuals with early-stage amyloid-related disease, including [mention specific example].

[0308] 14A. The method described in Embodiment 13A, wherein the individual is suspected of having an amyloid-related disease.

[0309] 15A.a. Quantifying amyloid in an individual by administering an amyloid-reactive peptide containing the amino acid sequence described in one of SEQ ID NOs: 1-14, conjugated to a detectable label, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs. b. To provide treatment for amyloid-related diseases. c. Administer an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs: 1-14, conjugated to a detectable label, to an individual, and quantify amyloid in the individual at a second time point by detecting the amount of the detectable label in the individual's tissues or organs, and d. To determine whether the treatment is effective by comparing the amyloid quantified at the first time point in the tissue or organ with the amyloid quantified at the second time point in the same tissue or organ. A method for monitoring the treatment of amyloid-related disease in individuals with early-stage amyloid-related disease, including [specific example].

[0310] 16A. The method according to any one of Embodiments 1A to 15A, further comprising administering an amyloid-reactive peptide containing an amino acid sequence described in any one of Sequence IDs 1 to 14 conjugated to a detectable label to an individual, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs, thereby quantifying amyloid in the individual at a second time point, optionally a third time point, a fourth time point, and / or a fifth time point.

[0311] 17A. The method according to embodiment 15A or 16A, wherein the first time point and the second time point are separated by at least six weeks.

[0312] 18A. A method for selecting a treatment for amyloid-related disease in an individual, the method being: a. Administering an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs. 1-14, conjugated to a detectable label, to an individual, and b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual. The method comprising, wherein if amyloid is detected in the heart, treatment for amyloid-related disease is performed, and if amyloid is not detected in the heart, alternative therapy is performed.

[0313] 19A.a. Quantifying amyloid in an individual at a first time point by administering an amyloid-reactive peptide containing the amino acid sequence described in one of SEQ ID NOs: 1-14, conjugated to a detectable label, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs. b. To provide treatment for amyloid-related diseases. c. Administer an amyloid-reactive peptide containing the amino acid sequence described in one of SEQ ID NOs: 1-14, conjugated to a detectable label, to an individual, and quantify the amount of amyloid in the individual at a second time point by detecting the amount of the detectable label in the individual's tissues or organs. d. Comparing amyloid quantified at a first time point in a tissue or organ with amyloid quantified at a second time point in the tissue or organ, and e. Adjusting treatment for amyloid-related diseases. A method for managing the treatment of amyloid-related diseases in an individual, including the treatment of amyloid-related diseases in an individual.

[0314] 20A. The method according to Embodiment 19A, wherein the treatment is modified if the amyloid disease is stable between the first and second time points.

[0315] 21A. The method according to embodiment 20A, wherein treatment is resumed if the amyloid load is increased.

[0316] 22A. The method according to any one of Embodiments 9A to 12A and 14A to 21A, wherein the amount of amyloid-reactive peptide exceeding a threshold indicates that an individual has an amyloid-related disease.

[0317] 23A. The method according to Embodiment 22A, wherein the amyloid-related disease is an early stage of amyloid-related disease.

[0318] 24A. The method according to any one of Embodiments 1A to 23A, wherein the amyloid-reactive peptide comprises SEQ ID NO: 13.

[0319] 25A. The method according to any one of Embodiments 1A to 24A, wherein the detectable label is a radioactive label.

[0320] 26A. Detectable labels are 11 C, 18 F, 111 In, 99m Tc, 89 Zr, 68 Ga and 123 I, 124 I and 125 A method according to any one of Embodiments 1A to 25A, selected from the group consisting of I.

[0321] 27A. Detectable labels 124 I is the method described in any one of Embodiments 1A to 26A.

[0322] 28A. Doses of approximately 0.1 to 25 mCi 124 A method according to any one of Embodiments 1A to 27A, comprising administering an I-labeled amyloid-reactive peptide to an individual.

[0323] 29A. Doses of approximately 0.3 mCi, 1 mCi, or approximately 2 mCi 124 A method according to any one of Embodiments 1A to 28A, comprising administering an I-labeled amyloid-reactive peptide to an individual.

[0324] 30A. A dose of approximately 1 mCi (±10%) 124A method according to any one of Embodiments 1A to 29A, comprising administering an I-labeled amyloid-reactive peptide to an individual.

[0325] 31A. A method according to any one of Embodiments 1A to 30A, comprising administering a dose of Tc-99 of approximately 20 mCi to an individual.

[0326] 32A. A method according to any one of Embodiments 1A to 31A, comprising administering approximately 0.3 mg, 1 mg, 1.5 mg, or approximately 2 mg of amyloid-reactive peptide to an individual.

[0327] 33A. The method according to any one of Embodiments 1A to 32A, wherein an amyloid-reactive peptide is administered intravenously.

[0328] 34A. The method according to any one of Embodiments 1A to 33A, further comprising determining organ-specific standard uptake ratios for organs within an individual.

[0329] 35A. The method according to Embodiment 34A, wherein the organ-specific SUVR for an individual is selected from the group consisting of mean SUVR, maximum SUVR, and peak SUVR.

[0330] 36A. The method according to any one of Embodiments 1A to 35A, further comprising determining organ-specific SUV for an individual.

[0331] 37A. The method according to Embodiment 36A, wherein the organ-specific SUV for an individual is selected from the group consisting of SUV mean, SUV maximum, and SUV peak.

[0332] 38A. The method according to any one of Embodiments 1A to 37A, further comprising determining the organ-specific dose per gram percentage (%ID) for an individual.

[0333] 39A. The method according to any one of Embodiments 1A to 38A, wherein the sensitivity for detecting amyloid using an amyloid-reactive peptide is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%.

[0334] 40A. A method according to any one of Embodiments 1A to 39A, wherein the sensitivity is approximately 99% or approximately 100%.

[0335] 41A. The method according to any one of Embodiments 1A to 40A, wherein the specificity of the amyloid-reactive peptide is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%.

[0336] 42A. The method according to any one of Embodiments 1A to 41A, wherein the specificity of the amyloid-reactive peptide is approximately 99% or 100%.

[0337] 43A. The method according to any one of Embodiments 1A to 42A, further comprising determining one or more health-related quality of life indicators for an individual.

[0338] 44A. The method according to any one of Embodiments 1A to 43A, further comprising detecting one or more biomarkers associated with amyloid-related disease.

[0339] 45A. The method according to Embodiment 44A, wherein the biomarker associated with amyloid-related disease is selected from the group consisting of troponin T, NTproBNP, urinary protein levels, UACR, EGFR, and alkaline phosphatase levels.

[0340] 46A. The method according to any one of Embodiments 1A to 45A, wherein the tissue or organ of the individual is selected from the group consisting of lungs, fat, heart, kidneys, pancreas, joints, spine, liver, spleen, adrenal gland, bone lesions, choroid plexus, pituitary gland, uterus, bone marrow, musculoskeletal tissue, gastrointestinal tract, and prostate.

[0341] 47A. The method according to any one of Embodiments 1A to 8A, 13A and 22A to 46A, wherein the threshold is the SUVR average threshold.

[0342] 48A. If the organ or tissue is the liver, the mean SUVR threshold is approximately 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, or 1.6; if the organ or tissue is the spleen, the mean SUVR threshold is approximately 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5; if the organ or tissue is the kidney, the mean SUVR threshold is approximately 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, or 2.3; and if the organ or tissue is the pancreas, The method according to Embodiment 47A, wherein the mean SUVR threshold is approximately 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, or 1.7, and if the organ or tissue is the heart, the mean SUVR threshold is approximately 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, or 2.1, or if the organ or tissue is the adrenal gland, the mean SUVR threshold is approximately 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, or 1.4.

[0343] 49A. The method according to Embodiment 48A, wherein the mean SUVR threshold is 1.31 when the organ or tissue is the liver, 1.21 when the organ or tissue is the spleen, 1.88 when the organ or tissue is the kidney, 1.40 when the organ or tissue is the pancreas, 1.19 when the organ or tissue is the heart, or 1.35 when the organ or tissue is the adrenal gland.

[0344] 50A. The method according to any one of Embodiments 1A to 8A, 13A and 22A to 46A, wherein the threshold is the SUVR maximum threshold.

[0345] 51A. If the organ or tissue is the liver, the maximum SUVR threshold is 10.6, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5 or 16; if the organ or tissue is the spleen, the maximum SUVR threshold is 3.8, 4, 4.5, 5, 5.5 or 5.9; and if the organ or tissue is the kidney, the maximum SUVR threshold is 7.7, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5 or 1 The method according to Embodiment 50A, wherein the maximum SUVR threshold is 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11 or 11.4 if the organ or tissue is the pancreas, 4.9, 5, 5.5, 6, 6.5, 7 or 7.5 if the organ or tissue is the heart, or 1, 1.1, 1.2, 1.3, 1.4, 1.5 or 1.6 if the organ or tissue is the adrenal gland.

[0346] 52A. The method according to Embodiment 51A, wherein the maximum SUVR threshold is 13.26 when the organ or tissue is the liver, 4.86 when the organ or tissue is the spleen, 9.68 when the organ or tissue is the kidney, 9.48 when the organ or tissue is the pancreas, 6.20 when the organ or tissue is the heart, or 1.31 when the organ or tissue is the adrenal gland.

[0347] 53A. The method according to any one of Embodiments 1A to 52A, further comprising performing a cardiac biopsy if amyloid is detected in the heart.

[0348] 54A. The method according to any one of Embodiments 1A to 53A, further comprising performing additional amyloid imaging on the individual.

[0349] 55A. The method according to any one of Embodiments 1A to 54A, further comprising performing additional amyloid imaging on the individual before administering the amyloid-reactive peptide.

[0350] 56A. The method according to Embodiment 55A, wherein additional amyloid imaging of the individual includes ECHO, CMR, bone scintigraphy, or positron emission tomography imaging.

[0351] 57A. Additional amyloid imaging of the individual 99m Tc-PyP, 99m Tc-DPD, 99m Tc-HMDP, 99m Tc-MDP, other bone scintigraphy tracers, 124 I and 18 F-Florbetapyr, 18 F-Frutemtamol and 18 The method according to embodiment 56A, further comprising a tracer selected from the group consisting of F-fluorbetabene.

[0352] 58A. The method according to any one of Embodiments 1A to 57A, wherein the diagnosis, prognosis, or response is confirmed by an indicator of the individual's health-related quality of life, the presence or amount of one or more biomarkers associated with amyloid-related disease, cardiac biopsy, and / or additional amyloid imaging of the individual.

[0353] 59A. A method according to any one of Embodiments 1A to 58A, wherein an individual is diagnosed with amyloid cardiomyopathy.

[0354] 60A. The method according to any one of Embodiments 1A to 59A, wherein the individual is suspected of having amyloid cardiomyopathy.

[0355] 61A. The method according to any one of Embodiments 1A to 60A, wherein the amyloid-related disease is systemic or localized amyloidosis.

[0356] 62A. The method according to any one of Embodiments 1A to 61A, wherein the amyloid-related disease is cardiac amyloidosis.

[0357] 63A. The method according to any one of Embodiments 1A to 62A, wherein the amyloid-reactive peptide has pan-amyloid specificity.

[0358] 64A. Amyloid-reactive peptides include immunoglobulin light chain (AL), immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin (ATTR wild-type; ATTR mutant), apolipoprotein AI (AApoAI), apolipoprotein AII (AApoAII), apolipoprotein AIV (AApoAIV), gelzolin (AGel), apolipoprotein C-II (AApoCII), apolipoprotein C-III (AApoCIII), lysozyme (ALys), and leukocytes. A method according to any one of Embodiments 1A to 63A, which binds to a chemotactic factor (ALECT2), fibrinogen α variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP); α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), IAAP, ALβ4, or ALβ1 amyloid.

[0359] 65A. The method according to any one of Embodiments 1A to 64A, wherein amyloid-reactive peptides are detected using PET, PET / CT, gamma machinography, SPECT, and / or SPECT / CT.

[0360] 66A. The method according to any one of Embodiments 1A to 65A, wherein the amyloid-related disease is selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AApoCII, AApoCIII, AGel, Alys, ALECT2, AFib, ACys, ACa, AMed, AIAPP, APro, AIns, APrP, and Aβ amyloidosis.

[0361] 67A. The method according to embodiment 66A, wherein the amyloid-related disease is cardiac AL amyloidosis.

[0362] 68A. The method according to embodiment 66A, wherein the amyloid-related disease is cardiac ATTR amyloidosis.

[0363] 69A. The method according to any one of Embodiments 1A to 68A, wherein the individual has a genetic predisposition to amyloid-related disease.

[0364] 70A. The method according to any one of Embodiments 1A to 69A, wherein the individual has a family history of amyloid-related disease.

[0365] 71A. The method according to any one of Embodiments 1A to 70A, wherein the individual is elderly.

[0366] 72A. The method according to any one of Embodiments 1A to 71A, wherein the individual has an early-stage amyloid-related disease.

[0367] 73A. The method according to any one of Embodiments 1A to 72A, wherein the individual has early-stage systemic amyloidosis.

[0368] 74A. The method according to any one of Embodiments 1A to 73A, wherein the individual has early-stage cardiac amyloidosis.

[0369] 75A. The method according to any one of Embodiments 1A to 74A, wherein the individual has early-stage AL amyloidosis.

[0370] 76A. The method according to Embodiment 75A, wherein early-stage AL amyloidosis is diagnosed according to the Mayo Clinic system.

[0371] 77A. The method according to embodiment 76A, wherein the early stage AL amyloidosis is stage 1 AL amyloidosis.

[0372] 78A. The method according to any one of Embodiments 1A to 77A, wherein the individual has early-stage ATTR amyloidosis.

[0373] 79A. The method according to Embodiment 78A, wherein early-stage ATTR amyloidosis includes stage 1 ATTR amyloidosis.

[0374] 80A. The method according to any one of Embodiments 1A-12A, 16A-18A, and 20A-79A, further comprising providing treatment for amyloid-related disease.

[0375] 81A. The method according to any one of Embodiments 13A to 15A, 19A and 80A, wherein the treatment for amyloid-related disease is selected from the group consisting of transthyretin stabilizers (e.g., tafamidis, acolamidis), transthyretin silencers (e.g., patisirane, inotercene, butricirane, eprotercene), gene editing approaches, anti-amyloid approaches utilizing monoclonal antibodies, therapies targeting plasma cell clones (e.g., daratumumab, bortezomib), and antibody-peptide fusions comprising an antibody-peptide fusion protein containing a second amyloid-reactive peptide and an antibody that binds to amyloid fibril.

[0376] 82A. The method according to Embodiment 81A, wherein the second amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 14, comprising 0, 1, 2, 3, or 4 amino acid substitutions, insertions, or deletions.

[0377] 83A. The method according to Embodiment 81A or 82A, wherein the antibody-peptide fusion protein comprises a heavy chain and a light chain, and an amyloid-reactive peptide is ligated at the C-terminus of the antibody's light chain via a spacer.

[0378] 84A. Antibody-peptide fusion protein, (i) an amyloid-reactive peptide containing the amino acid sequence described in Sequence ID No. 2, and (ii) Antibodies that bind to human amyloid fibrils The method according to any one of Embodiments 81A to 83A, wherein the antibody comprises a heavy chain and a light chain, the heavy chain of the antibody comprises a heavy chain variable region (VH), and the light chain of the antibody comprises a light chain variable region (VL), wherein VH comprises CDR-H1 comprising the amino acid sequence described in SEQ ID NO: 106, CDR-H2 comprising the amino acid sequence described in SEQ ID NO: 107, and CDR-H3 comprising the amino acid sequence described in SEQ ID NO: 108, and VL comprises CDR-L1 comprising the amino acid sequence described in SEQ ID NO: 103, CDR-L2 comprising the amino acid sequence described in SEQ ID NO: 104, and CDR-L3 comprising the amino acid sequence described in SEQ ID NO: 105, wherein the amyloid-reactive peptide and the antibody are linked at the C-terminus of the light chain, and the amyloid-reactive peptide is linked to the antibody via a spacer comprising the amino acid sequence described in SEQ ID NO: 83.

[0379] 85A. A method according to any one of Embodiments 1A to 84A, wherein a peptide containing an amyloid-reactive peptide and an N-terminal leader sequence is administered to an individual.

[0380] 86A. The method according to Embodiment 85A, wherein the peptide containing the amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO: 100.

[0381] 87A. The method according to Embodiment 86A, wherein a detectable label is bound to one or more amino acids at positions 1 to 6 of the amino acid sequence described in Sequence ID No. 100.

[0382] 88A. Detectable labels 124 The method described in Embodiment 87A, which is I.

[0383] 89A. 124 The method according to Embodiment 88A, wherein I is bound to amino acid 4 of SEQ ID NO: 100.

[0384] 90A. The method according to any one of Embodiments 1A to 89A, further comprising administering potassium iodide to a solid.

[0385] 91A. The method according to Embodiment 90A, wherein the administration of potassium iodide is performed by administering 130 mg of potassium iodide to the patient for 7 days, starting one day before the administration of the amyloid-reactive peptide.

[0386] 92A. The method according to Embodiment 91A, wherein the administration of potassium iodide is performed by administering 130 mg of potassium iodide to the patient for 3 days, starting approximately 30 minutes before the administration of the amyloid-reactive peptide.

[0387] 93A. The method according to any one of Embodiments 90A to 92A, wherein potassium iodide is administered orally.

[0388] 94A. A peptide comprising the amino acids described in Sequence ID No. 100, wherein the peptide comprises a detectable label bound to one or more amino acids at positions 1 to 6.

[0389] 95A. Detectable signs 124 Peptide I, according to Embodiment 94A.

[0390] 96A. 124 The peptide according to Embodiment 95A, wherein I is bound to amino acid 4 of SEQ ID NO: 100.

[0391] Examples The following embodiments are described solely for illustrative purposes and are not intended to limit the scope of this disclosure.

[0392] This disclosure is not intended to be limited in scope to any particular embodiment disclosed, which is described, for example, to illustrate various aspects of this disclosure. Various modifications to the compositions and methods described will become apparent from the description and teachings herein. Such modifications are achievable without departing from the true scope and spirit of this disclosure and are intended to be included within the scope of this disclosure.

[0393] Example 1: 124 Cardiac amyloid detection by PET / CT imaging of I-p5+14 This example describes the safety and efficacy of a novel amyloid-reactive peptide, referred to as p5+14, labeled with iodine-124, in patients with various types of systemic amyloidosis.

[0394] Test design This is in patients with any type of systemic amyloidosis, pre-symptomatic subjects with germline TTR mutations, and healthy subjects. 124 This was a multi-part, open-label trial to evaluate the safety, dosimetry, and biodistribution of the I-p5+14 peptide. In Part 5, one patient received repeated imaging at approximately two-year intervals (Table 5). Patients with systemic AL amyloidosis were initially recruited to evaluate radiodosemetry before enrolling additional patients. Due to the early termination of this trial to allow for the transfer of the IND to a commercial partner for clinical development, the enrollment target was not met (Table 5).

[0395] This trial is for the safe and accurate detection of amyloid deposition in patients with systemic amyloidosis as an imaging agent. 124 This was an exploratory Phase 1 / Ph2 evaluation of I-p5+14. The primary objective was to assess the safety and tolerability of the study drug using systemic and organ-specific dose measurements (Part 1) as well as standard safety indicators (vital signs, adverse events, and laboratory indicators).

[0396] Secondary objectives were broadly designed to evaluate the following: 1. In all patients with systemic amyloidosis, and within each major amyloid type patient subset, e.g., AL, ATTR, ALECT2, in anatomical sites where biopsy-proven or clinically suspected amyloid deposition was present, 124 Is it possible to observe organ-specific retention of I-p5+14?

[0397] 2. In patients who can be appropriately evaluated, PET data-based assessment of renal and cardiac conditions 124 Correlation between I-p5+14 uptake (defined as mean and maximum concentration per unit volume [Bq / cc] or SUVR) and clinical metrics of organ function such as serum NT proBNP, creatinine, BUN, or eGFR.

[0398] 3. Subjects without abnormal amyloid deposition, i.e., healthy controls 124 Distribution of I-p5+14.

[0399] 4. In asymptomatic subjects with known pro-amyloid mutations in transthyretin 124 In vivo distribution of I-p5+14.

[0400] 5. Can changes in amyloid load in patients be recognized through repeated imaging?

[0401] This trial enrolled n=50 patients with diagnosed and identified types of systemic amyloidosis, 2 asymptomatic subjects with identified variant TTR at risk of amyloidosis (these were not evaluable and were excluded from organ-based and patient-based sensitivity measurements), and 5 healthy controls (a total of 57 unique subjects). In addition, one participant from Part 2 was included. 124 Participants successfully underwent repeated exposure to I-p5+14 and follow-up imaging. Participants were not randomized or blinded for their disease status. All subjects 124The patient was administered the I-p5+14 PET imaging agent. During the evaluation of PET / CT image data, the experimental nuclear medicine specialist (radiologist) was blinded to the diagnosis and suspected organ lesions of the patient or subject.

[0402] This study consisted of five parts. In Part 1 (n=3 patients with extensive AL amyloidosis), patients participated in a radioactivity dose escalation study to evaluate radiation dose measurement and pharmacokinetics. 124 Patients received a single intravenous (IV) bolus of I-p5+14 (less than 2 mg of p5+14 peptide). Patient 1 received 0.31 mCi (0.84 mg p5+14), Patient 2 received 1.05 mCi (1.06 mg p5+14), and Patient 3 received 1.03 mCi (0.95 mg p5+14). Whole-body and organ-specific dose measurements were estimated based on imaging data using the OLINDA-EXM method (1).

[0403] In parts 2, 3, and 4, we registered an additional 54 participants. 124 I-p5+14(1.98±0.19mCi) 124 A single intravenous (IV) infusion of 1 and 1.42 ± 0.19 mg of p5+14) was administered. In Part 5, one patient who was imaged during Part 2, 124 I underwent a second infusion of I-p5+14 (2.08 mCi and 1.49 mg p5+14) and repeated PET / CT scans.

[0404] Radiolabeled drug products were synthesized using aseptic techniques in a biosafety cabinet within a certified cleanroom at the Radiochemistry Facility in Knoxville, University of Tennessee Medical Center. After testing for radiochemical purity, stability, biological activity, and endotoxin levels, the products were mixed in a sterile phosphate buffer of 35 mL or less containing 5% w / v human serum albumin (USP) as a stabilizer and 0.5 mg / mL ascorbic acid (USP) as an antioxidant. 124 The I-p5+14 dosage was formulated. Participants received 30 mL delivered at a flow rate of 3 mL / min. 124 I-p5+14 was administered intravenously (IV), followed by a 30 mL saline flush at the same rate. Iodine-124 was purchased from 3D Imaging (Little Rock, AR) and prepared according to the information provided in DMF#025853. [Table 4]

[0405] Patient identification and imaging protocols To confirm the diagnosis of amyloidosis and evaluate clinical evidence for organ lesions in 13 predetermined organs and tissues, this study required an initial pre-screening of medical records after obtaining signed medical release consent (Figure 1). Following written consent, on day 1, participants underwent a physical examination and blood draw (venotomy), followed by the initiation of a 7-day course of oral potassium iodide administration. Subjects received 3 mL / min 124 Thirty minutes prior to the 30 mL intravenous (IV) infusion of I-p5+14, the patient received an administration of acetaminophen and an antihistamine, followed by a 30 mL saline flush at the same rate.

[0406] Blood samples were taken again on day 3. Follow-up telephone calls were made on days 9 and 28 for patients with systemic amyloidosis and pre-symptomatic subjects. Healthy subjects recruited after 42 patients had been imaged underwent safety follow-up on days 3, 9, 28, and 56, and provided blood samples on days 28 and 56 (Figure 1).

[0407] Subject population Participants were recruited locally and from across the United States. Patients included men and women over 18 years of age with a diagnosis of a known type of amyloidosis. Healthy subjects were over 50 years of age, without sex or race restrictions, without known germline mutations in the TTR gene, without a diagnosis of amyloidosis or type 2 diabetes, and without first- or second-degree relatives with confirmed or suspected familial amyloidosis. One patient with evidence of AL and positive organ uptake underwent repeated imaging approximately 24 months later.

[0408] Due to known interactions between heparin and peptide p5+14, patients exposed to heparin within 7 days prior to infusion were excluded.

[0409] A total of n=25 patients with AL amyloidosis (n=23 with systemic AL and n=2 with focal AL); n=20 patients with ATTR amyloidosis (n=15 with ATTRv and n=5 with ATTRwt); 2 patients (n=2) with ALECT2 amyloidosis; and (n=3) patients with other amyloid types were imaged (Table 3). In addition, n=5 healthy volunteers and n=2 asymptomatic TTRv carriers were enrolled. [Table 5] TIFF2026533995000007.tif64138

[0410] Synthesis of radioactive tracers and PET / CT imaging Peptide p5+14 was synthesized by AmbioPharm Inc. (North Augusta, SC) and supplied as 3 mg aliquots of lyophilized powder, stored at -20°C. Iodine-124 was purchased from 3D Imaging (Little Rock, AR; DMF# 025853). Patient batches of radioactive tracers were prepared on the day of use at the UT radiochemical facility using soluble iodogen as an oxidizing agent. Images were acquired using a Siemens Biograph PET / CT (Siemens, Knoxville, TN) with approximately 30 seconds of low-dose CT (120 kVp, effective 50 mAs). PET images were first acquired from the knee to the ankle using a 2 x 10 minute bed position, followed by imaging from the crown of the head to the thigh using a 5 minute bed position. Finally, a 10 minute gated cardiac PET image was acquired.

[0411] PET data were reconstructed using a 3D OSEM algorithm with attenuation and prompt gamma-ray correction, a 168×168 image matrix, and an image resolution of approximately 8 mm full width at half maximum. CT data were reconstructed using a moderate smoothing kernel and a 4 mm reconstruction increment.

[0412] Image interpretation PET / CT images were visually evaluated for uptake in 13 organs and tissues using the XD General Oncology Review application in Mirada Medical DBx (Build 1.2.0.59). Maximum Intensity Projection (MIP) and PET / CT images were created using Inveon Research Workplace (IRW) software (Ed. 4.2 [4.2.0.15], Siemens Preclinical Solutions). PET / CT image evaluation was performed by radiologists blinded to clinical data. 124I-p5+14 uptake was manually quantified by measuring radioactivity per unit volume (Bq / cc). The standard uptake ratio (SUVR) was determined for all organs by scaling the mean radioactivity concentration by the blood pool (radioactivity in the lumen of the thoracic aorta).

[0413] In patients with AL or ATTR-related amyloidosis, 124 The correlation between cardiac uptake of I-p5+14 (mean SUVR) and serum NT-proBNP at the same time was evaluated. Similar correlations were examined for renal radiotracer uptake, as well as for serum urea nitrogen (BUN), creatinine, and eGFR.

[0414] endpoint The primary endpoint was radiation dose measurement and safety determination based on the collection of adverse events (AEs), which were reported according to the MedDRA (Medical Dictionary for Regulatory Activities) after initial classification using the CTCAE criteria. Additional efficacy-based secondary and exploratory endpoints, as well as two indicators of interest, were also initiated.

[0415] evaluation Other clinical evaluations besides serum biomarker measurements were not performed as part of this study. Safety evaluations were conducted as follows: 124 These tests were performed before and after I-p5+14 infusion. Vital signs were collected immediately before and for 50 minutes thereafter, along with the infusion of the radioactive tracer. Blood samples were taken for clinical chemistry, complete blood count, C-reactive protein, lactate dehydrogenase, and NTproBNP testing (Figure 1). In healthy subjects, additional blood samples were collected at 28 and 56 days post-infusion to evaluate anti-p5+14 antibodies using in-house peptide capture ELISA.

[0416] Estimation of Amyloid Organ Positivity The true standards of diagnosis and organ lesions in this study were derived from medical records and based on standard diagnostic methods. Tissue biopsies were performed in most patients. Organ lesions were tallied during pre-screening. Cardiac amyloidosis was generally based on a combination of serum biomarkers (NT-proBNP), echocardiography, magnetic resonance imaging, nuclear medicine imaging, and endocardial biopsy. Amyloid lesions in other organs relied on a combination of organ-specific or distant biopsy and serum biomarkers.

[0417] statistics The sample size was determined by the rarity of systemic amyloidosis in the population, the expected cumulative rate, and the exploratory nature of the study. Given the number of subjects, a comprehensive subset analysis of patients with AL and ATTR, as well as healthy volunteers, was not feasible. All subjects who completed the study were included in the image analysis, with the exception of pre-symptomatic participants whose image quality was deemed unacceptable.

[0418] PET / CT imaging of abdominal and thoracic organs 124 Patient-based and organ-based sensitivity was determined by comparing the visual uptake of I-p5+14 with clinically known or expected organ lesions. Sensitivity and specificity (percent agreement) were calculated using the exact method, and 95% confidence intervals (95% CI) were calculated using the Wilson-Brown method (Graphpad Prism v9.3.0). Patient-based sensitivity (i.e., positive in any organ or tissue) was determined. 124 Participants with a confirmed diagnosis of amyloidosis accompanied by I-p5+14 uptake were similarly calculated. Cardiac specificity was evaluated in a cohort of healthy subjects where the absence of amyloid was assumed based on the recruitment criteria.

[0419] Cardiac data in patients and healthy subjects 124 Comparison of I-p5+14 uptake was performed using an unpaired two-sided t-test with α=0.05. In the kidney or heart... 124The correlation between I-p5+14 uptake (SUVR mean) and BUN (mg / dL), creatinine (mg / dL), eGFR, and serum NT-proBNP (pg / mL) was determined using Spearman analysis (α=0.05). Calculations were performed using SPSS Version 28 (Armonk, NY: IBM Corp.) and Prism.

[0420] ethical supervision The trial protocol was approved by the U.S. Food and Drug Administration and conducted with the support of an Investigational New Drug (IND) submission (IND# 132282). Further approval was obtained from the Institutional Review Board (IRB) at the UT Graduate School of Medicine (Knoxville, TN). All patients provided written consent regarding pre-screening of medical records and imaging protocols prior to participation in the trial.

[0421] result Breakdown / Trends of Participants Fifty-seven subjects (n=57) were enrolled (Table 4). For the purposes of most analyses, healthy volunteers (n=5) and asymptomatic TTRv carriers (n=2) constituted the amyloid-free control group. The first three patients were administered 11.4 MBq, 38.9 MBq, and 38.1 MBq, respectively, and were imaged seven times over 48 hours post-infusion. Subsequent subjects (n=54) were administered 73.3 ± 7.0 MBq and 1.42 ± 0.19 mg of peptide, and PET / CT images were acquired 5.2 hours post-infusion (IQ, 4.93–5.42). The first n=22 patients were also imaged approximately 24 hours post-infusion, but this was discontinued to minimize CT radiation dose as the data did not provide additional information for interpreting the findings at 5 hours post-infusion. All subjects completed the study.

[0422] Patient background and baseline characteristics 51 percent (51%) of the subjects were male. The mean age of all subjects was 64.8 ± 10.2 years. In the AL amyloidosis population, 52% (n=13 / 25) were λ isotype and 40% (n=10 / 25) were κ isotype. 8 percent (n=2 / 25) were unreported (Table 5). Patients with ATTR amyloidosis included 25% ATTRwt (n=5 / 20). The ATTRv population included eight different germline mutations, the majority of which were T60A (n=6 / 20) (Table 5). Cardiac lesions were clinically observed in 52% (n=13 / 25) and 55% (n=11 / 20) of patients with AL or ATTR amyloidosis, respectively. Cardiac amyloidosis was clinically expected in only one patient among those with other types of amyloidosis (ALys). In the AL patient cohort, extracardiac amyloid was found in medical records in the spleen (n=1 / 25), liver (n=3 / 25), and kidney (n=7 / 25).

[0423] Manufacturing of radioactive tracers 124 Each batch of I-p5+14 was evaluated for reactivity with amyloid-like fibrils, and the mean percentage of binding peptides was equal at 97.5 ± 1.5% in the prepared doses. 124 The average radiochemical purity of I-p5+14 was 98.8 ± 0.7%. All other release criteria, specifically pyrogenicity, sterility, color, pH, peptide identity, peptide purity, and particulate matter, met the release criteria.

[0424] Dosimetry and pharmacokinetics The average whole-body effective radiation dose for each sex was estimated to be 0.23 (±0.02) mSv / MBq, with the highest organ dose received by the bladder (0.94 ± 0.25 mSv / MBq). Radioactivity excretion (elimination) occurred primarily via the renal and gastrointestinal pathways, accompanied by rapid loss of radioactivity from the central compartment. The estimated whole-blood elimination half-life was 21.9 ± 7.6 hours.

[0425] In healthy subjects 124Cardiac uptake and abdominal / thoracic biodistribution of I-p5+14 In healthy subjects (n=5), radioactivity was observed in the stomach and bladder, as well as in saliva from the parotid and salivary glands, renal pelvis, ureters, and esophagus. In healthy subject #4, radioactivity in the liver and kidneys was considered higher than the blood pooled background, and consequently, these sites were considered positive (Figure 2A). In the heart, no visible evidence of radioactivity was found in the LV or right ventricular (RV) wall or IVS in any of the healthy subjects (Figure 2B). Therefore, 124 Visual evaluation of the I-p5+14 distribution showed a specificity (negative percentage agreement) of 100% for the heart and spleen, and 80% for the liver and kidney (Table 5). [Table 6]

[0426] In patients 124 Cardiac uptake of I-p5+14 124 Cardiac uptake of I-p5+14 was readily visible by PET / CT imaging in patients with ATTR or AL amyloidosis (Figure 3A), as well as in patients with AApoA1 and ALys-related amyloidosis. Cardiac uptake of the radiotracer was observed in patients with known cardiac lesions and in patients without identified cardiac lesions (Figure 3B). In most cases, radioactivity was observed in the LV wall and IVS, associated with diverse lesions in the RV wall and atria.

[0427] heart 124 The sensitivity for I-p5+14 uptake was 93% (n=13 / 14, [95% CI: 68.5-99.6]) and 100% (n=11 / 11, [95% CI: 74.2-100]) in patients with AL and ATTR amyloidosis, respectively. For the overall patient population, cardiac sensitivity was 96% (n=25 / 26, [95% CI: 81.1-99.8]) (Table 6). 124Cardiac retention of I-p5+14 was observed in 27% (n=3 / 11) of AL patients and 67% (n=6 / 9) of ATTR patients who did not have clinical evidence of cardiac lesions based on a review of medical records. Gated cardiac imaging was not evaluated. [Table 7] TIFF2026533995000010.tif205150

[0428] heart 124 Quantitative analysis of I-p5+14 uptake and correlation with serum NTproBNP. No difference was observed in mean cardiac SUVR between the AL patient cohort and the ATTR patient cohort (Figure 4A). Cardiac uptake of radioactive tracers was significantly higher in patients with systemic amyloidosis than in healthy subjects (Figure 4A). Mean cardiac SUVR was significantly higher in the PET-positive (PET+) cohort than in PET-negative (PET-) patients (Figure 4A).

[0429] In patients with AL amyloidosis, a statistically significant correlation (rS=0.48, p=0.018) was observed between mean cardiac SUVR and serum NTproBNP (Figure 4B). No correlation was observed in patients with ATTR amyloidosis (rS=0.28, p=0.231) (Figure 4C). In the post-hoc analysis of AL amyloidosis patient data, mean cardiac SUVR was scaled by serum creatinine for each patient to account for diverse renal function impairments. A very significant correlation (rS=0.61, p=0.0017) was obtained when comparing this composite variable with serum NTproBNP (Figure 4D). No correlation was observed between mean renal SUVR and serum creatinine, BUN, or eGFR.

[0430] In patients 124 Extracardiac intracellular distribution of I-p5+14 124I-p5+14 binding was visually observed in one or more of the n=13 pre-determined organs in 94% of registered patients (n=44 / 47, [95% CI: 82.8-97.8]) (Table 6). Two patients diagnosed with AL (n=2) and one patient diagnosed with ATTR showed positive visual adhesion in any of the organs or tissues evaluated. 124 It did not have Ip5+14 acquisition. 124 Extracardiac connectivity of I-p5+14 was observed in the kidneys, liver, spleen, pancreas, lungs, adrenal glands, bones and joints, subcutaneous and abdominal fat, pituitary gland, choroid plexus, and bone lesions (in patients with multiple myeloma and AL amyloidosis) (Figures 5A-5B). Despite the small number of clinical observations, the sensitivity for hepatic and splenic amyloid in all patients was 100% (n=4 / 4, 95% CI: 51.0-100) and 100% (n=2 / 2, 95% CI: 17.8-100) (Table 6). The kidney, as an excretory organ, had a lower sensitivity of 79% (n=11 / 14, 95% CI: 52.4-92.4) (Table 6). 124 I-p5+14 uptake was also observed in the spleen and liver in 37.5% (n=9 / 24) and 18% (n=4 / 22), respectively, of patients without clinical evidence of organ-related lesions. 124 I-p5+14 uptake was observed in the spleen (35%, n=7 / 20), liver (24%, n=5 / 20), and kidney (45%, n=9 / 20) of patients with ATTR amyloidosis in the absence of clinical evidence of organ disease.

[0431] Immunogenicity of p5+14 In all samples, there was no evidence of anti-peptide antibodies in ELISA when compared to two control samples of normal human serum.

[0432] safety Safety analysis was performed on all subjects. Overall, 124I-p5+14 was well tolerated, and there were no deaths, serious drug-related adverse events (SAEs), or treatment-related adverse events (TEAEs) leading to discontinuation of the study (Table 8). Only one patient (1.8%, n=1 / 57) experienced drug-related TEAEs, namely pruritus and dysuria, both of which resolved without sequelae. Approximately half of the subjects (49.1%, n=28 / 57) reported drug-unrelated TEAEs, most of which were mild. [Table 8]

[0433] Consideration Current anatomical imaging techniques, including echocardiography and cardiac magnetic resonance imaging, provide valuable structural and functional information after a diagnosis of amyloidosis has been confirmed. 99m Tc-PyP and similar osteoaffin-facing agents have been used to detect cardiac amyloid. However, they appear to bind to microcalcifications associated with amyloid deposition (Dietemann S, Nkoulou R. Ann Nucl Med. 2019;33:624-628), and their relationship with amyloid fibrils is not fully understood. Therefore, non-invasive imaging methods using reagents that directly bind to amyloid deposits, are quantifiable, and can detect cardiac amyloidosis before the onset of obvious anatomical changes and organ dysfunction would offer significant clinical benefits.

[0434] Peptide p5+14 is a synthetic peptide with pattern recognition ability that binds to amyloid via multivalent electrostatic interactions (Antoni G, Lubberink M, Estrada S, et al. J Nucl Med. 2013;54:213-220). In preclinical studies, the peptide bound to synthetic amyloid-like fibrils and tissue sections containing various types of amyloid. When radiolabeled with iodine-124, the peptide is suitable for PET / CT imaging, providing quantitative, high-resolution in vivo distribution data. Iodine-124 is a non-persistent radionuclide produced in a cyclotron with a half-life of 4.2 days, and is enzymatically cleaved from the peptide by intracellular enzymes during renal catabolism, leading to its physiological uptake in the gastric lumen, salivary glands, and thyroid gland (Cuddy SAM, Bravo PE, Falk RH, et al. JACC Cardiovasc Imaging. 2020;13:1325-1336). This process is used to enable amyloid imaging in the kidney, a clearance organ.

[0435] this is, 124 This was the first Phase 1 / 2 trial of I-p5+14 in humans, enrolling patients with eight different types (subtypes) of amyloidosis: ALκ, ALλ, ATTRwt, ATTRv, Alys, AGel, ALECT2, and AApoA1. The sensitivity of organ-specific uptake of the radiotracer was evaluated by comparison with data in the patients' medical records (which were used as the true standard in this early-stage trial). Incidental imaging findings were also recorded after visual examination of PET images. Overall, 124 The 2mCi dose of I-p5+14 (approximately 1.4 mg of peptide) was well tolerated, and no deaths or drug-related SAEs were observed. The 1mCi dose used in subsequent studies... 124The whole-body effective dose (8.9 mSv) from I-p5+14 injection is comparable to numerous thallium-201 and technetium-99m cardiac evaluations (Genovesi D, Vergaro G, Giorgetti A et al., JACC Cardiovasc Imaging. 2021;14:246-255). With the exception of one AL patient diagnosed with cutaneous amyloid nodules and one patient with renal amyloidosis who responded well to treatment and was presumed to have achieved complete regression of amyloid deposition, all patients diagnosed with amyloidosis in this study had at least one abdominal or thoracic organ or tissue, including the heart. 124 Visual uptake of I-p5+14 was observed (18,19). One patient diagnosed with systemic ATTRwt amyloidosis with peripheral neuropathy and Congo red-positive ligamentum flavum findings tested negative.

[0436] Cardiac uptake of radioactive tracers was observed in 100% of ATTR patients whose clinical evidence supported a diagnosis of cardiac amyloidosis. Furthermore, positive cardiac PET / CT imaging was observed in six ATTR patients without clinical evidence of cardiac lesions. 99m The study included patients with Tc-PyP imaging and normal serum NTproBNP levels. These data were obtained from: 124 PET / CT imaging of I-p5+14 suggests that it may be possible to detect early, pre-symptomatic cardiac amyloid.

[0437] Among the major abdominal and thoracic organs, clinical findings and 124 The sensitivity (positive percentage agreement) with I-p5+14 imaging was strongest for the heart (96.2%; 95% CI: 80.4-99.9; n=26). The presence of cardiac amyloid was not confirmed concurrently and relied on clinical reports in medical records, which is a limitation of the study design, but it suggests that cardiac amyloid persists even in patients responding to treatment. PET / CT imaging revealed the external cardiac region. 124Incidental uptake of I-p5+14 has been observed, particularly in the liver and spleen of patients with AL amyloidosis, which are organs that generally contain asymptomatic amyloid deposits (Martin EB, Williams A, Richey T et al., Sci Rep. 2016;6:22695). ATTR amyloidosis is primarily considered a cardiac or neurological disease, but it is a systemic pathology. In patients with ATTR amyloidosis... 124 PET / CT imaging of I-p5+14 showed diverse anatomical uptake, including, for example, the lungs, liver, kidneys, pancreas, ball-and-socket joints, spine, and abdominal fat. These data showed that 124 I-p5+14 imaging suggests that it can not only detect cardiac amyloidosis but also extracardiac diseases that may contribute to a reduced quality of life and affect the rate of cardiac amyloid removal in patients receiving amyloid-degrading drugs.

[0438] PET imaging is inherently a quantitative modality. 124 I-p5+14 uptake can be readily assessed by various analytical methods, including the determination of organ-specific SUVR, a semi-quantitative indicator in PET. This ratio was used in this study to adjust for differences in the clearance rates of radiotracers. Clearance rates can be significantly affected by renal impairment, particularly in patients with AL amyloidosis and in a large number of relatively rare types with ubiquitous renal lesions. In this study, mean cardiac SUVR measurements were indistinguishable between the AL and ATTR populations. However, the AL, ATTR, and other amyloid populations had significantly higher mean cardiac SUVRs compared to a small cohort of healthy controls.

[0439] Serum NT-proBNP is a validated biomarker for overall survival in patients with AL amyloidosis. 124Cardiac retention of I-p5+14 showed a positive and significant correlation with measurements taken at the same time, but not in ATTR patients. This correlation increased when renal function, estimated by serum creatinine levels, was included in the analysis (Dorbala S, Ando Y, Bokhari S et al., J Nucl Cardiol. 2020;27:659-673).

[0440] Current amyloid treatments have been shown to have a greater impact in patients with cardiac amyloidosis in the earlier stages of the disease; therefore, early diagnosis of the disease is essential. Substances that can directly detect amyloid deposition before the onset of symptoms or when symptoms are mild can be advantageous. As new amyloid removal therapies are developed, means of directly measuring amyloid load, quantifying changes, and demonstrating effectiveness will become extremely valuable. 124 Non-invasive and quantitative techniques that can detect and monitor cardiac and extracardiac depositions, such as PET / CT imaging using I-p5+14, would be particularly useful.

[0441] conclusion Data from this initial human trial will enable the detection of not only systemic amyloid deposition but also cardiac amyloidosis. 124 This supports the overall safety and efficacy of I-p5+14. Based on these data, orphan drug designation for the diagnosis of AL and ATTR amyloidosis was granted in the United States and Europe.

[0442] Example 2: Inclusion and Exclusion Criteria This embodiment describes the inclusion and exclusion criteria for the tests disclosed in Example 1.

[0443] The following inclusion and exclusion criteria were used in the study.

[0444] 1. The patient had a confirmed diagnosis of systemic amyloidosis (SA) based on either histological confirmation of biopsy containing deposits of apple-green birefringent Congo-red affinity material, genetic screening and the presence of amyloid-related pathology, or amyloid-specific imaging studies. The type of amyloidosis (AL, ATTR, ALECT2, or other) was also characterized.

[0445] 2. Patients enrolled in Part 1 (n=3) had extensive AL amyloidosis, defined as biopsy-proven or clinically detectable lesions in at least two organs (excluding the peripheral nervous system).

[0446] 3. All patients in Parts 1 and 2 were 18 years of age or older. There were no restrictions based on gender or race.

[0447] 4. Women of childbearing potential (those who have not been surgically sterilized, who are not postmenopausal [typically understood to mean more than two years have passed since their last menstrual period without the intervention of medication], and who are capable of conception) had to test negative for pregnancy in clinical tests performed by the facility's physician.

[0448] 5. Patients who have previously received treatment, are currently receiving treatment, or are taking other drug-based anti-amyloid regimens were eligible to participate in the trial (Parts 1 and 2).

[0449] 6. The patient provided signed written informed consent and complied with eligibility requirements, scheduled visits, and follow-up studies.

[0450] 7. Due to annual dose measurement (exposure) limitations, patients who have previously participated in another nuclear medicine amyloid imaging clinical trial protocol are eligible to enroll in this trial only after at least 12 months have passed since their previous radiotracer injection.

[0451] Prior to participation in Part 3 (n=2) of this examination, the following inclusion criteria were required: 1. The patient had a clear germline mutation in the transthyretin (TTR) gene, which is considered to be at risk for amyloidosis, but had no clinical evidence of SA.

[0452] 2. All patients in Part 3 were over 50 years old. There were no restrictions based on gender or race.

[0453] 3. Women of childbearing potential (those who have not been surgically sterilized, who are not postmenopausal [typically understood to mean more than two years have passed since their last menstrual period without the intervention of medication], and who are capable of conception) had to test negative for pregnancy in clinical tests performed by the facility's physician.

[0454] 4. Participants provided signed written informed consent and complied with eligibility requirements, scheduled visits, and follow-up studies.

[0455] 5. Due to annual dose measurement limitations, subjects who have previously participated in another nuclear medicine amyloid imaging clinical trial protocol are eligible to enroll in this trial only after at least 12 months have passed since their previous radiotracer injection.

[0456] Prior to participating in Part 4 of the exam (n=5), the following inclusion criteria were required: 1. Healthy control subjects (HCs) were generally healthy adults, either male or female, who had no diagnosis of amyloidosis, no first or second-degree relatives (parents, siblings, children, aunts, uncles, nieces, nephews) with confirmed or suspected familial amyloidosis, and did not have type 2 diabetes mellitus.

[0457] 2. All Part 4 subjects were 30 years of age or older.

[0458] 3. Women of childbearing potential (those who have not been surgically sterilized, who are not postmenopausal [typically understood to mean more than two years have passed since their last menstrual period without the intervention of medication], and who are capable of conception) had to test negative for pregnancy in clinical tests performed by the facility's physician.

[0459] 4. The patient provided signed written informed consent and complied with eligibility requirements, scheduled visits, and follow-up studies.

[0460] 5. Due to annual dose measurement limitations, patients who have previously participated in another nuclear medicine imaging clinical trial protocol are eligible to enroll in this trial only after at least 12 months have passed since their last radiotracer injection.

[0461] Prior to taking Part 5 of the exam (n=1), the following inclusion criteria were required: 1. The patient has successfully completed Part 2 or Part 3 of this clinical trial, and the patient's CT / PET images demonstrate the presence of abnormal amyloid deposition in the abdominal and thoracic organs.

[0462] 2. Patients receive a dose of 2 mCi during Part 2 or 3 of this trial. 124 The patient had received I-p5+14 and had visual evidence of radioactive tracer uptake in amyloid-related abdominal and thoracic organs.

[0463] 3. Repeated exposure to the test drug was performed at least 6 months after the initial exposure.

[0464] 4. Patients submitted serum samples to exploratoryally assess the presence of p5+14 peptide-reactive antibodies, and the results were reviewed by the principal investigator prior to a second exposure to the study drug.

[0465] 5. The patient met all inclusion criteria for participation in Part 2 or Part 3, including signing informed consent regarding Part 5.

[0466] The following criteria excluded candidates from participating in Parts 1 and 2 of the exam: 1. Individuals with significant comorbidities (e.g., an Eastern Cooperative Oncology Group [ECOG] score of 3 or higher), uncontrolled infections, or other serious illnesses.

[0467] 2. Patients with a sustained SpO2 of 92% or less, as recorded in their medical records.

[0468] 3. Patients who required dialysis.

[0469] 4. Pregnant or breastfeeding women of childbearing potential who have tested positive for pregnancy in a clinical examination conducted by a facility physician (women who have not been surgically sterilized, are not postmenopausal [typically understood to mean more than two years have passed since their last menstrual period without the intervention of medication], and are of fertile capacity).

[0470] 5. Patients who have received any amyloid-affinity radiotrace within the past 12 months as part of an investigational clinical trial (not standard treatment).

[0471] 6. Patients who have been exposed to heparin or heparin-based drugs within 7 days prior to the imaging study.

[0472] 7. Patients with a known allergy to acetaminophen or iOSAT iodine therapy.

[0473] The following criteria excluded candidates from participating in Part 3 of the exam: 1. Individuals with significant comorbidities (e.g., an Eastern Community Oncology Group [ECOG] score of 3 or higher), uncontrolled infections, or other serious illnesses.

[0474] 2. Subjects with a sustained SpO2 of 92% or less, as recorded in their medical records.

[0475] 3. Pregnant or breastfeeding women of childbearing potential who have tested positive for pregnancy in a clinical examination conducted by a facility physician (women who have not been surgically sterilized, are not postmenopausal [typically understood to mean more than two years have passed since their last menstrual period without the intervention of medication], and are of fertile capacity).

[0476] 4. Subjects who had clinical evidence of amyloidosis based on standard clinical criteria.

[0477] 5. Subjects with multiple neurological disorders of unknown cause.

[0478] 6. Subjects who, within the past 12 months, received any amyloid-affinity radiotrace as part of an investigational clinical trial (not standard treatment).

[0479] 7. Subjects who have been exposed to heparin or heparin-based medications within 7 days prior to the imaging study.

[0480] 8. Subjects with a known allergy to acetaminophen or iOSAT iodine treatment.

[0481] 9. Subjects with clinical signs of SA based on routine serum and urine biomarker studies, radiological or nuclear medicine imaging studies, or peripheral nerve evaluation.

[0482] The following criteria excluded subjects from participation in Part 4: 1. Individuals with significant comorbidities (e.g., an Eastern Community Oncology Group [ECOG] score of 2 or higher (meaning the individual is able to walk and perform light or sedentary work, such as light housework or office work, but is physically limited to strenuous activity)), uncontrolled infections, or other serious illnesses.

[0483] 2. Individuals with a sustained SpO2 of 92% or less, as recorded in their medical records.

[0484] 3. Pregnant or breastfeeding women of childbearing potential who have tested positive for pregnancy in a clinical examination conducted by a facility physician (women who have not been surgically sterilized, are not postmenopausal [typically understood to mean more than two years have passed since their last menstrual period without the intervention of medication], and are of fertile capacity).

[0485] 4. Subjects who have received any amyloid-affinity radiotrace as part of an investigational clinical trial (not standard treatment) within the past 12 months.

[0486] 5. Subjects who have been exposed to heparin or heparin-based drugs within 30 days prior to the imaging study.

[0487] 6. Subjects with a known allergy to acetaminophen or iOSAT iodine therapy.

[0488] 7. Subjects who had been diagnosed with type 2 diabetes or were taking medication to manage type 2 diabetes.

[0489] 8. An active infection requiring systemic antiviral or antimicrobial therapy that is not completed by day 1 of the trial.

[0490] 9. A known history of or positive test result for human immunodeficiency virus (HIV), hepatitis C, or chronic hepatitis.

[0491] 10. Uncontrolled hypertension (blood pressure > 160 / 100 mmHg).

[0492] 11. Smoking more than 20 cigarettes a day.

[0493] 12. Diagnosis of left ventricular ejection fraction-preserving heart failure.

[0494] 13. In the opinion of the principal investigator, there were any other conditions that could make the subject unsuitable for inclusion or prevent the subject from participating in or completing the study.

[0495] The following criteria excluded subjects from participation in Part 5: 1. Any patient who has experienced a clinically significant adverse event related to prior exposure to the study drug.

[0496] 2. Any subject who meets either of the exclusion criteria for Part 2 or Part 3 (if applicable).

[0497] Example 3: 124 I-P5+14( 124 Cardiac amyloid quantification using I-p5+14) versus 18F-florbetapil: Pilot PET / CT study This example shows that in participants with amyloid cardiomyopathy (CMP) and controls, the myocardium 124 To quantify I-p5+14 uptake and its diagnostic value 18 This document describes a pilot study aimed at comparing F-florbetapir with F-florbetapir. The primary objective of this pilot study was to (1) compare participants with amyloid CMP and controls. 124 (2) Quantifying I-p5+14 cardiac uptake in AL-CMP and ATTRwt-CMP 124 Ip5+14 and 18 To compare the diagnostic value of F-florbetapir, and (3) 124 The objective was to measure the correlation between I-p5+14 myocardial uptake and indicators of cardiac structure and function, serum biomarkers, functional status, and quality of life as markers of amyloid load.

[0498] method Participant inclusion This trial was approved by the Mass General Brigham Human Research Committee, and each participant provided written informed consent (protocol number 2021P000085). 124The use of I-p5+14 was approved by the Food and Drug Administration under Investigational New Drug Application (155808). Between July 2021 and June 2023, the following 46 participants were enrolled: 12 AL-CMPs, 12 ATTRwt-CMPs, 2 hereditary amyloid CMPs (1 mutant ATTR [ATTRv] with Thr60Ala [p.T80A], 1 apolipoprotein A-IV amyloidosis [AApoAIV]), and 12 control participants without amyloidosis. 18 Eight control participants from a previous study using F-florbetapyr PET / CT were included (Dorbala S, Vangala D, Semer J, et al. Eur J Nucl Med Mol Imaging. 2014;41:1652-1662). One AL-CMP participant was, 124 I-p5+14 was excluded due to acquisition errors during imaging. Participants with hereditary amyloidosis were not included in the statistical analysis due to their small number (N=2), but were visually presented for comparison. AL-CMP was diagnosed using standard criteria for systemic AL amyloidosis, including evidence of cardiac lesions by biopsy and imaging or endocardial biopsy with confirmation of amyloid type by immunohistochemistry or mass spectrometry. ATTRwt amyloidosis was defined as grade 2 / 3 cardiac amyloidosis. 99m Tc-pyrophosphate ( 99m AL amyloidosis was diagnosed by exclusion of AL amyloidosis by Tc-PYP)SPECT / CT scan and serum free light chain assay and immunofixed electrophoresis of serum and urine, or by endocardial biopsy with immunohistochemistry or mass spectrometry. Hereditary amyloid CMP was diagnosed by biopsy with immunohistochemistry or mass spectrometry, detection of mutant TTRs by gene sequencing as needed, and evidence of cardiac lesions by imaging or endocardial biopsy.

[0499] PET / CT acquisition Participants with amyloid CMP were 124 I-p5+14 and 18 I underwent molecular imaging of amyloidosis using F-florbetapyr PET / CT.124 (Excluding one AL-CMP participant for whom only I-p5+14 PET / CT was available). The controls underwent one PET / CT: 12 participants 124 I-p5+14, 8 people 18 F-Florbetapir. Imaging was performed using a Discovery MI PET / CT scanner (GE Healthcare, Chicago, IL, USA) with CT scouting for patient positioning and low-dose chest CT scans for attenuation correction and image fusion. Acquisition and quantification methods were specific to each radiotracer, taking into account differences in radiotracer characteristics. 124 I-p5+14 was intravenously infused over 5 minutes. Cardiac PET acquisition was performed 5 hours after infusion for 30 minutes in static mode only, to allow clearance of pooled blood radioactivity, based on previous data regarding radiotracer kinetics. The median net infused radioactivity, limited by concerns regarding exposure to participants and staff, was 1.00 mCi (interquartile range [IQR] 0.94–1.07), and the median effective dose was 9.01 mSv (IQR 8.55–9.47, including 0.5 mSv from a low-dose CT scan, based on 0.23 mSv / MBq). The inventors protected the thyroid with 130 mg of potassium iodide for 7 days, starting 1 day prior to radiotracer infusion. 18F-florbetapil was injected as a bolus 1 minute after the start of PET acquisition in list mode. Static images of the heart were reconstructed using data from 4 to 30 minutes after radioactive tracer injection, based on previous publications. The median net injected radioactivity was 7.15 mCi (IQR 6.33–8.40), and the median effective dose was 5.58 mSv (IQR 4.99–6.46, including 0.5 mSv from a low-dose CT scan). 124 The I-p5+14 scan is, 18 The scan was performed a median of 1.5 days after the F-florbetapir scan (IQR 1-4.5, range 1-12 days).

[0500] PET / CT quantitative Myocardial uptake was measured volumetrically on static images using PMOD software (PMOD Technologies LLC, Zurich, Switzerland). Because manual tracing of myocardial contours on each slice was unreliable and inaccurate in controls, the inventors used automated iso-contouring in PMOD to exclude the ventricular blood pool and depict the myocardium in a reproducible manner. First, the region of interest (VOI) was manually traced on PET emission images guided by fused CT images to define the contours of the left ventricle (LV) and right ventricle (RV), including the lumen blood pool, with the interventricular septum assigned to the LV trace. A specific contour threshold was selected for each radiotracer, taking into account differences in radiotracer dynamics and target-to-background ratios. Blood pool radioactivity concentrations were measured in a 10 mm diameter left atrial (LA) spherical VOI. 18 For F-florbetapyr, a threshold of 2 × mean pooled blood radioactivity was used, as in our previous studies. 124 For I-p5+14, we used a threshold of mean + 2 standard deviations of the blood pooled radioactivity concentration, which better depicted myocardial volume and was particularly noticeable in mild cases (Figure 6).

[0501] 124 To perform automated contour plotting on I-ebzamitide PET images, the inventors first manually traced a region of interest (VOI) on a fused PET / CT image to define the overall contours of the left and right ventricles (LV and RV). Next, they measured the blood pooled radioactivity concentration in a 10 mm diameter left atrial (LA) spherical VOI. Then, they used the contour plotting function of the PMOD software with a threshold of mean + 2 standard deviations (SD) of the LA blood pooled radioactivity concentration, which adequately depicted myocardial volume in cases with varying levels of uptake. In contrast, in the last column, 18 A threshold of 2 × mean LA blood pool radioactivity concentration, as used in F-florbetapil, would ignore the relevant portion of myocardial volume. This is why the inventors 124This explains why a specific threshold (LA+2SD) was used for I-ebzamitide.

[0502] The inventors' primary LV and RV uptake indices were percentage of injected radioactivity (%ID), calculated as VOI mean radioactivity concentration × VOI volume / injected radioactivity. This indice is adjusted for injected radioactivity but not for body weight, the latter being unnecessary for radiotracers that accumulate in the heart and specific organs rather than the whole body. The inventors further analyzed standardized uptake values ​​(SUV mean, SUV maximum) as mean or maximum VOI radioactivity concentration / (injected radioactivity / body weight), cardiac amyloid radioactivity (CAA) as VOI SUV mean × VOI volume, and target-to-background ratio (TBR) as VOI mean radioactivity concentration / blood pool mean radioactivity concentration (identical to the SUV ratio, where corrections for injected radioactivity and body weight in SUV are offset in the ratio). The threshold for abnormally high uptake values ​​was defined by maximizing the Euden index (sensitivity + specificity - 1) in receiver operating characteristic (ROC) analysis to identify the cutoff with optimal sensitivity and specificity for classifying CMP cases versus controls, giving equal weight to false-positive and false-negative cases.

[0503] Structural and functional markers of amyloid loading Reports and, where available, images from echocardiograms (N=23), cardiac MRIs (N=13), and 99mTc-PYP SPECTs (N=11) performed clinically within one year of PET / CT were retrieved. Older 99mTc-PYP SPECT results were also included if they were grade 3, as no further changes were anticipated. The median time lag to PET / CT was 1.7 months (IQR 0.7–4.1) for echocardiograms, 2.7 months (IQR 0.7–5.0) for MRI, and 4.5 months (IQR 2.5–13.8) for SPECT. Ventricular septal thickness, LV mass index, ejection fraction, myocardial contractility (MCF = stroke volume / myocardial volume), global longitudinal strain (GLS), delayed gadolinium enhancement, ECV, and 99mTc-PYP grade were assessed.

[0504] Clinical tests, functional status, and quality of life Serum levels of troponin T, N-terminal pro B-type natriuretic peptide (NT-proBNP), and creatinine were measured in all participants. Functional status was assessed using the New York Heart Association classification (NYHA) and the 6-minute walk distance. Quality of life was measured using the Minnesota Living with Heart Failure Questionnaire (MLWHFQ; 21 questions, total score 0-105 points, higher scores indicate worse symptoms), the Kansas City Cardiomyopathy Questionnaire (KCCQ; 23 questions, total score 0-100 points, higher scores indicate less symptoms), and the 36-Item Short Form Survey (SF-36; 36 questions, T-score 0-100 converted for a normalized population, higher scores indicate better quality of life).

[0505] statistical analysis Continuous variables were presented as medians with IQRs and compared using the Kruskal-Wallis test, followed by Dunn tests for pairwise comparisons. Multiple comparison adjustments were performed using the Benjamini-Hochberg method. Categorical variables were presented as frequencies with percentages and evaluated using Fisher's exact test. Paired variables were compared using Wilcoxon signed-rank tests or McNemar tests as needed. Correlations were quantified using Spearman's ρ with 95% confidence intervals (CI) after filler correction. Two-tailed p-values ​​were presented, and values ​​less than 0.05 were considered statistically significant. Data were analyzed using R version 4.3.0 (R Core Team.R: A language and environment for statistical computing.R Foundation for Statistical computing, Vienna, Austria) with the tidyverse, DescTools, gtsummary, rstatix, pROC, and correlation packages.

[0506] result Patient characteristics In the amyloid CMP participants (N=24), the median age was 74 years (IQR 69–78), and 22 were male (92%). ATTRwt-CMP participants (N=12) were older than AL-CMP participants (N=12), had lower heart rates, and poorer cardiac metrics such as LV mass index, ejection fraction, and GLS. However, other demographics, clinical features, quality of life, functional status, and biomarkers were similar (Table 9). At trial inclusion, 10 AL-CMP participants were in hematological remission after plasma cell targeted therapy (83%), 2 were undergoing treatment and not in remission (17%), while 10 ATTRwt-CMP participants were receiving tafamidis (83%). In the control group (N=20), the median age was 62 years (IQR 58–67), and 13 were male (65%). [Table 9] TIFF2026533995000013.tif143163

[0507] 124 I-P5+14 PET / CT for LV and RV myocardial uptake Radioactive tracer injection was well tolerated in all participants, and no adverse effects were observed. Regardless of amyloid type, in all amyloid CMP participants, 124 Significant cardiac uptake was observed in I-p5+14, but none of the control participants showed visible cardiac uptake. Furthermore, grade 1 indeterminate 99m In participants who showed Tc-PYP SPECT / CT, 124 I-p5+14 PET / CT showed clear myocardial uptake, which was confirmed by endocardial biopsy showing TTR amyloid deposition (Figure 7). 124 I-p5+14 LV %ID differed between groups (overall p<0.001): the median for AL-CMP was 1.48 (IQR 1.12–1.89), for ATTRwt-CMP 2.12 (1.66–2.47), and for controls 0.00 (0.00–0.01, Table 8). Several uptake indices tended to be higher in ATTRwt-CMP than in AL-CMP, particularly in the mean LV SUV and RV indices (Figure 8, Table 10). Based on the Euden index, high LV %ID, LV CAA, and LV TBR completely differentiated amyloid CMP cases from controls, while the mean LV SUV, maximum SUV, and RV indices showed some overlap (Figure 8, Table 10). Indeed, several control participants exhibited heterogeneous background radioactivity with small spots of radioactivity concentrations exceeding the autocontour threshold based on blood pooling. As a result, the average SUV fell within the range for amyloid CMP, but it did not visually suggest amyloid deposition. In contrast, %ID and CAA were close to zero in the control participants because these indicators include VOI volume and therefore better differentiated amyloid deposition in CMP cases from heterogeneous background radioactivity in the control participants. [Table 10] TIFF2026533995000015.tif32162

[0508] 18 LV and RV myocardial uptake by F-fluorbetapyr PET / CT Same amyloid CMP participant ( 18 In a different control participant (excluding one AL-CMP participant in whom F-florbetapyr PET / CT was not performed), the inventors found that, 124 Similar to I-p5+14 18 Similar results were obtained with F-florbetapyr (Figure 9, Table 10). 18 F-florbetapir LV %ID also differed between groups (overall p<0.001): the median was 1.03 (IQR 0.78–2.03) for AL-CMP, 1.10 (0.89–1.44) for ATTRwt-CMP, and 0.04 (0.00–0.29) for controls. LV myocardial uptake indices did not differ between AL-CMP and ATTRwt-CMP participants, although the range of values ​​was higher for AL-CMP. High LV %ID and LV CAA completely differentiated amyloid CMP cases from controls, while mean LV SUV, LV TBR, and RV indices showed some overlap.

[0509] 124 I-P5+14 pairs 18 F-Florbetapyr-based myocardial radioactive tracer uptake index When comparing LV myocardial radiotracer uptake within the same participants, the %ID, SUV mean, SUV maximum, and CAA for LV and RV were as follows in AL-CMP: 124 Ip5+14 and 18 It was similar to F-fluorbetapyr, but in ATTRwt-CMP, 124 I-p5+14 was higher (all p ≤ 0.002, Figure 10, Table 10). LV and RV TBR were higher in AL-CMP and ATTRwt-CMP. 124 I-p5+14 18F-florbetapir showed higher results (all p ≤ 0.002), which reflects the different hemodynamics of the two tracers.

[0510] 124 Correlation between I-P5+14 uptake and direct and indirect markers of amyloid load, functional status, and quality of life. Across the entire cohort, the inventors stated that 124 We found moderate correlations between I-p5+14 LV %ID and NYHA classification (ρ=0.47), MLWHFQ total score (ρ=0.59), KCCQ overall score (ρ=-0.62), SF-36 physical T score (ρ=-0.50), and SF-36 mental T score (ρ=-0.41), as well as strong correlations with troponin T (ρ=0.73) and NT-proBNP (ρ=0.72) (Table 9). Furthermore, in AL-CMP and ATTRwt-CMP participants, we found that, 124 I-p5+14 LV %ID and, 18 Moderate to strong correlations were found between F-florbetapir LV %ID (ρ=0.50), ventricular septal thickness (echocardiogram ρ=0.78, MRI ρ=0.58), LV mass index (echocardiogram ρ=0.70, MRI ρ=0.82), LV MCF (echocardiogram ρ=-0.66, MRI ρ=-0.62), LV GLS (ρ=0.54), and LV ECV (ρ=0.51). [Table 11]

[0511] Consideration This pilot study involves a novel panamyloid-binding radiotracer in participants with amyloid CMP. 124 The myocardial uptake of I-p5+14 was quantified, and its diagnostic value was compared with that of 18F-florbetapil in amyloid CMP. 124 We measured the correlation between I-p5+14 myocardial uptake and markers of amyloid load. 124Visual interpretation of I-p5+14 PET / CT images identified cardiac amyloidosis in all participants with AL, ATTRwt, ATTRv, or AApoAIV CMP. 124 Metrics (indicators) of I-p5+14 LV and RV myocardial quantitative uptake were significantly higher in participants with either AL-CMP or ATTRwt-CMP than in control participants. Importantly, 124 I-p5+14 LV %ID, CAA, and TBR completely differentiated all known cases of AL-CMP and ATTRwt-CMP from the control group. Similar results were observed with 18F-florbetapil, except for TBR, in the same amyloid CMP participants. 124 This supports the validity of the findings for I-p5+14. Therefore, LV %ID and CAA were the most accurate metrics for identifying amyloid CMP in both radiotracers. When comparing radiotracers, with the exception of TBR, 124 I-p5+14 consistently showed higher myocardial values ​​in ATTRwt-CMP. Furthermore, among AL-CMP and ATTRwt-CMP participants, 124 I-p5+14 LV %ID correlates moderately to strongly with indirect metrics of amyloid load from echocardiography and MRI (ventricular septal thickness, LV mass index, MCF, GLS, ECV), and reflects a reasonable quantification of amyloid load. Furthermore, 124 I-p5+14 LV %ID correlates moderately with 18F-florbetapyr LV %ID, reflecting different binding sites, different binding affinities, and / or the quantification of different components of amyloid deposition. In fact, 124 I-p5+14 is specifically designed to bind to the electrically negative surface of glycosaminoglycans and amyloid fibrils, while 18F-florbetapyr is a stilbene, structurally similar to thioflavin T, which is known to bind to the β-sheet surface along channels formed by cross-strand ladders within amyloid fibrils. 124The correlation between I-p5+14 LV %ID and ECV was also moderate, which may be due to ECV measuring fibrosis, inflammation, and / or edema in addition to amyloid deposition. The correlation with functional status and quality of life was moderate, and the correlation with cardiac biomarkers was strong, which suggests that 124 This further supports the validity of cardiac amyloid quantification using I-p5+14 PET / CT. Finally, 124 I-p5+14 infusion was well tolerated by all participants, and no adverse effects were observed.

[0512] 124 I-p5+14 is an innovative radiotracer based on the synthetic polypeptide p5+14, developed to bind to the electrically negative surface of glycosaminoglycans and amyloid fibrils in all types of amyloid deposition. This polypeptide has been successfully tested in a mouse model of serum amyloid A amyloidosis and in a variety of radiotracers in humans with multiple types of systemic amyloidosis. This study was conducted in a cohort of participants with amyloid CMPs derived from multiple precursor proteins. 124 It is novel in that it allows for the systematic evaluation of myocardial uptake using I-p5+14, and also 18 This is the first comparison with F-florbetapyr. Our findings contribute to the accurate and safe diagnosis of AL-CMP and ATTRwt-CMP, as well as to the identification of hereditary amyloid CMP. 124 The use of I-p5+14 PET / CT imaging is supported. These results suggest that this pan-amyloid-binding radiotracer may reduce the need for endocardial biopsy to diagnose cardiac amyloidosis, potentially limiting biopsies to scan-positive patients where confirmation of amyloid type is necessary. Our results also support the use of I-p5+14 PET / CT imaging. 124 This suggests that I-p5+14 can quantify cardiac amyloid load. In fact, LV 124I-p5+14 uptake correlated with structural and functional LV indices of amyloid load, as well as cardiac biomarkers and heart failure-related QOL (quality of life) before treatment. 124 Imaging with I-p5+14 allowed for the identification of amyloid presence in the heart and other organs, quantification of amyloid levels, and specific prediction of potential major effects and organ-specific side effects. Post-treatment imaging, on the other hand, allowed for the measurement of organ responses. While ECV may be an alternative approach for quantifying amyloid load in the heart, it is not specific to amyloidosis and may represent inflammation or fibrosis (processes that can be induced by macrophage-mediated amyloid removal). Therefore, targeted imaging of cardiac amyloid using specific molecular radiotracers may offer significant advantages compared to other methods.

[0513] Recently, β-amyloid-targeting PET radiotracers [ 11 C-Pittsburgh-B compound, 18 F-Florbetapyr, 18 F-Fluorbetabene and 18 F-flutematamol has emerged as a useful means for evaluating AL-CMP. However, these radioactive tracers are not suitable for ATTR-CMP, especially 18 F-fluorbetapyr and 18F -The ability to detect florbetabene is limited. Non-invasive diagnosis of ATTR-CMP is usually performed using SPECT bone affinity radiotracers. 99m Tc-Pyrophosphate [ 99m Tc-PYP], 99m Tc-3-diphosphono-1,2-propanodicarboxylic acid [ 99m Tc-DPD], 99m Tc-hydroxymethylenediphosphonate [ 99mThis is based on Tc-HMDP). Grade 2 / 3 uptake of these radiotracers has been reported to be nearly 100% specific to ATTR-CMP, but with a sensitivity of only 74%. In fact, bone-affinity SPECT radiotracers may lack sensitivity for detecting early disease in ATTR-CMP and are unreliable for diagnosing AL-CMP, some forms of ATTRv-CMP, and other rare forms of amyloid CMP. Furthermore, these data were obtained from large centers evaluating selected patients with advanced disease or high pre-test probabilities, and therefore may overestimate the diagnostic performance of SPECT in non-specialized centers. Moreover, a series of quantitative tests using bone-affinity radiotracers are limited by the relatively low sensitivity of SPECT compared to PET and the early stages of development of quantitative SPECT techniques. Importantly, the mechanism of myocardial uptake of bone-affinity radiotracers is not publicly known, which impairs the interpretation of post-treatment changes. In contrast, PET panamyloid-binding radiotracers 124 I-p5+14 can provide higher quantitative accuracy than SPECT bone affinity radiotracers and may be more sensitive to ATTR-CMP than β-amyloid-specific PET radiotracers. In particular, in this study, the inventors observed that among ATTRwt-CMP participants, 18 F-florebetapil is better than 124 Significantly higher myocardial uptake was observed in I-p5+14. These participants also showed higher levels than AL participants. 124 While there was a tendency to show I-p5+14 uptake, uptake was more similar across amyloid types with 18F-florbetapir. In parallel, ATTRwt-CMP participants had worse structural and functional metrics, such as LV mass index and GLPS, on echocardiography and cardiac MRI than AL-CMP participants. Therefore, in ATTRwt-CMP... 124 Higher myocardial uptake values ​​measured by I-p5+14 may better reflect the severity of cardiomyopathy and may be useful in diagnosing the disease early.18 It may be more sensitive than F-florbetapir. There is a strong need for techniques that provide specific and quantitative estimations of both cardiac AL and ATTR amyloid loads to evaluate organ responses to amyloid removal therapy. 124 I-p5+14 has the potential to fill that gap.

[0514] in conclusion, 124 I-p5+14 PET / CT imaging is a promising novel radiotracer that can accurately detect multiple types of amyloid deposition, distinguish between the presence and absence of cardiac amyloidosis, and quantify cardiac amyloid load. In ATTRwt-CMP, 124 Amyloid quantification using I-p5+14 may be more accurate than amyloid quantification using 18F-florbetapil. These findings should be validated for the early diagnosis and therapeutic monitoring of cardiac amyloidosis. 124 To further evaluate the clinical usefulness of I-p5+14 PET / CT imaging, larger-scale, multicenter collaborative studies are needed.

[0515] Example 4: 124 Monitoring of amyloid progression and regression of I-p5+14 using PET / CT imaging. This example describes the use of a novel amyloid-reactive peptide called p5+14, labeled with iodine-124, in patients with various types of systemic amyloidosis. 124 This report describes the monitoring of amyloid progression and regression using semi-quantitative analysis of PET / CT imaging in I-p5+14).

[0516] method This applies to patients with any type of systemic amyloidosis, pre-symptomatic subjects with germline TTR mutations, and healthy subjects as described in Example 1. 124 This was a multipart, open-label study to evaluate the safety, dose measurement, and in vivo distribution of the I-p5+14 peptide. In this example, from baseline... 124To visualize and quantify changes in organ-specific uptake of the I-p5+14 peptide, nine AL patients and ten ATTR patients underwent repeated PET / CT imaging. The imaging protocol is described in Example 1.

[0517] By measuring the radioactivity per unit volume (Bq / cc) and standard uptake ratio (SUVR) scaled by blood pooling radioactivity, the radioactivity in organs can be assessed. 124 The uptake of I-p5+14 peptide was quantified. Correlations between serum NTproBNP, left ventricular (LV) wall and interventricular septum (IVS) thickness, and overall longitudinal strain and SUVR were calculated.

[0518] result In nine patients with AL amyloidosis, organ function was measured by standardized uptake ratio (SUVR) and serum NTproBNP. 124 Changes in I-p5+14 peptide uptake are shown in Figure 11A. In the AL patient population, 1 / 9 (11%) showed a decrease in mean SUVR to near normal, 7 / 9 (78%) showed a stable mean cardiac SUVR, 1 / 9 (11%) showed an increase in mean cardiac SUVR without a change in NTproBNP, 8 / 9 (89%) showed a stable NTproBNP, and 1 / 9 (11%) showed an increase in NTproBNP.

[0519] In nine patients with ATTR amyloidosis, organ function was measured by standardized uptake ratio (SUVR) and serum NTproBNP. 124 Changes in I-p5+14 peptide uptake are shown in Figure 11B. In the ATTR patient population, 2 / 10 (20%) showed a decrease of more than 20% in the mean cardiac SUVR, and 7 / 10 (70%) showed a stable mean SUVR. 1 / 10 (10%) showed an increase in the mean cardiac SUVR without a change in NTproBNP, 9 / 10 (90%) showed stable NTproBNP levels, and 1 / 10 (10%) showed a decrease in NTproBNP.

[0520] Across all patients, the change in NTproBNP between imaging time points was: 124 Changes in cardiac uptake in I-p5+14 correlated with these changes (Table 12, Figure 12). When each group was evaluated individually, changes in serum NTproBNP and in both AL patients and ATTR patients correlated. 124 No correlation was detected between changes in cardiac uptake for I-p5+14. [Table 12]

[0521] In AL patients, correlations were detected between SUVR and the LV wall and GLS, but no correlation was detected with IVS (Figure 13A-C). In ATTR patients, correlations were detected between SUVR and the LV wall and IVS, but no correlation was detected with GLS (Figure 13D-F). In all patients, correlations were detected between SUVR and the LV wall, IVS, and GLS (Figure 13G-I).

[0522] Consideration This example uses PET / CT imaging. 124 Semi-quantitative analysis of I-p5+14 uptake has shown that it enables monitoring of amyloid progression and regression, as well as a method for studying the relationship between amyloid, amyloidosis, organ function, and patient treatment outcomes.

[0523] Example 5: 124 Early cardiac amyloid detection by PET / CT imaging of I-p5+14 This study concerns the diagnosis of cardiac amyloidosis. 124 This is a Phase 3 evaluation of the efficacy of I-p5+14. The primary objective is to assess the diagnosis of cardiac amyloidosis based on visual scan interpretation. 124 The objective is to evaluate the sensitivity and specificity of PET / CT imaging using I-p5+14.

[0524] A secondary objective is to track the incidence of treatment-related adverse events (AEs) and changes from baseline in clinical laboratory values ​​from day 1 to day 30. 124 The objective is to evaluate the safety of a single intravenous administration of I-p5+14.

[0525] The exploratory objective is 1) the diagnosis of cardiac ATTR amyloidosis. 124 1) To evaluate the effectiveness of I-p5+14, and 2) Regarding the diagnosis of cardiac AL amyloidosis. 124 3) To evaluate the effectiveness of I-p5+14, and to differentiate between AL cardiac amyloidosis and ATTR cardiac amyloidosis. 124 To evaluate the effectiveness of I-p5+14, 4) 124 To evaluate the correlation between the quantitative cardiac uptake of I-p5+14 and amyloid content in endocardial biopsy, cardiac biomarkers, clinical stage, participant-reported heart failure health status, and standard structural and functional measurements using imaging, 5) Biopsy or multimodal imaging 124 The objective of I-p5+14 is to assess participants' preferences regarding PET / CT imaging.

[0526] Inclusion and exclusion criteria The following inclusion criteria will be used for this test.

[0527] 1) You must understand the test procedures and be able to provide signed informed consent (as defined in Section 10.1.3). This includes compliance with the Informed Consent Form (ICF) and the requirements and limitations described in this Protocol.

[0528] 2) A man or woman aged 18 or older.

[0529] 3) The participant is suspected of having cardiac amyloidosis and is willing to undergo diagnostic evaluation for cardiac amyloidosis (e.g., echocardiography, CMR, bone affinity tracer cardiac SPECT, extracardiac or endocardial cardiac biopsy). Participants may be enrolled before or during the diagnostic evaluation for cardiac amyloidosis.

[0530] 4) As part of the examination, the patient can undergo PET / CT imaging, including the ability to lie supine for approximately one hour.

[0531] 5) For women of childbearing age: During treatment and 124 You agree to maintain abstinence (refrain from heterosexual intercourse) or to use a method of contraception with an annual failure rate of less than 1% for at least 30 days after taking I-p5+14.

[0532] a. A woman is considered fertile if she has postmenarche, has not reached menopause (absence of 12 months or more without a specified cause other than menopause), and has not undergone surgical sterilization (removal of the ovaries and / or uterus). The definition of fertile may be adapted to align with local guidelines or requirements.

[0533] b. Examples of contraceptive methods with an annual failure rate of less than 1% include bilateral tubal ligation, male sterilization, established and appropriate use of oral contraceptives that suppress ovulation, hormone-releasing intrauterine devices (IUDs), and copper IUDs.

[0534] i. Any method of contraception with an annual failure rate of more than 1% is not permitted, such as spermicide-containing caps, diaphragms or sponges, or male or female condoms with or without spermicide.

[0535] ii. The reliability of sexual abstinence should be assessed in relation to the duration of the clinical trial and the participants' preferred normal lifestyle. Periodic abstinence (e.g., calendar method, ovulation method, symptomatic body temperature method, or post-ovulation method) and withdrawal are not acceptable methods of contraception.

[0536] 6) For men: agree to maintain abstinence or take contraceptive measures as defined below, and agree to refrain from donating sperm: a. If a man has a fertile female partner, he must abstain from sexual activity during the treatment period and for at least 120 days after the last dose of the investigational drug (30 days + 90 days of spermatogenesis cycles), or use condoms plus an additional method of contraception with a combined annual failure rate of less than 1%. The man must refrain from donating sperm during this same period.

[0537] The following exclusion criteria will be used in this test.

[0538] 1) Confirmed diagnosis of cardiac amyloidosis.

[0539] 2) Diagnosis of confirmed systemic amyloidosis (e.g., renal AL or ATTR peripheral neuropathy) without known cardiac lesions. Participants who are amyloid-positive only from carpal tunnel tissue or laminectomy are permitted unless they have other known organ lesions.

[0540] 3) At the time of enrollment, the patient is receiving treatment with an approved therapy (e.g., tafamidis) or is participating in a clinical trial for the treatment of cardiac ATTR amyloidosis. For AL and ATTR: Patients may enter the screening period for clinical trials for treatment after their ICF is signed and the participant is enrolled in this trial, but must not receive experimental therapy until after their visit for the 30-day safety follow-up in this trial.

[0541] 4) You are pregnant or breastfeeding.

[0542] 5) Being mentally or legally incapacitated, having significant emotional problems at the time of the examination, or having a history of mental illness.

[0543] 6) There is a known allergy to potassium iodide.

[0544] 7) Receiving hemodialysis or peritoneal dialysis.

[0545] 8) eGFR is less than 15 ml / min / 1.73m2.

[0546] 9) Myocardial infarction within 3 months of screening.

[0547] 10) Having severe claustrophobia or any medical condition that would prevent the completion of the imaging protocol.

[0548] 11) The principal investigator has a medical condition that could potentially interfere with the trial results or pose an additional risk to the participant.

[0549] 12) 124 Within 7 days prior to I-p5+14 administration, the patient received heparin or a heparin analog (e.g., enoxaparin, dalteparin, fondaparinux).

[0550] 13) Known, untreated thyroid disease (other than mild elevation of thyroid-stimulating hormone with normal T4 levels).

[0551] Subject population and identification This trial will enroll participants suspected of having cardiac amyloidosis. Enrollment will continue until at least 100 participants with positive cardiac amyloidosis (based on adjudication committee assessment) and 45 participants with negative cardiac amyloidosis (based on adjudication committee assessment) are enrolled, ensuring that the enrollment requirements for the primary endpoint are met. An independent clinical adjudication committee with expertise in cardiac amyloidosis will be established to review information obtained during the diagnostic evaluation (e.g., clinical history and laboratory tests, endocardial biopsy, extracardiac biopsy, echocardiography, cardiac MRI, and bone affinity tracer cardiac SPECT) to determine whether the data support a diagnosis of cardiac amyloidosis.

[0552] The screening period is a maximum of 30 days. Eligibility criteria are confirmed and informed consent is obtained. For women of childbearing potential (WOCBP), all WOCBPs are treated as follows: 124A serum pregnancy test (urgent serum β-hCG pregnancy test) is performed before administration of I-p5+14. The pregnancy test results are available and in all WOCBPs. 124 A negative result must be confirmed before administering I-p5+14.

[0553] The treatment period (period of symptom onset under treatment) is from day 1 to day 30. 124 If nuclear medicine imaging (e.g., SPECT with bone affinity tracer) is performed prior to I-p5+14 administration, the patient's visit on day 1 of the trial should take place at least 24 hours after the most recent nuclear medicine scan. 124 If a Tc-99m labeled SPECT scan is performed after an I-p5+14 PET / CT scan, 124 To minimize downscatter from I-p5+14 to SPECT scan, it may be necessary to wait until the fifth day visit or later.

[0554] The Standard of Truth (SoT) diagnostic evaluation period is for the diagnosis of cardiac amyloidosis. If all necessary diagnostic tests are not completed by day 30, the diagnostic evaluation period is extended until the last diagnostic test is completed or day 60, whichever comes first.

[0555] Test intervention and imaging protocols During the first day of the visit, the subjects presented with p5+14 at a dose level of 1 mCi (±10%) of 2 mg or less. 124 Receive a single dose of I-p5+14. 124 I-p5+14 is delivered intravenously by a push at a rate of approximately 1 mL / 5 seconds.

[0556] 124 Before administering I-p5+14: 1) Regarding all WOCBP, 124 1) A serum pregnancy test (urgent serum β-hCG pregnancy test) is performed before administration of I-p5+14 and a negative result is confirmed. 2) Participants are required to reduce the uptake of radioactive iodine in the thyroid gland. 1241) Start oral therapy with 130 mg of KI (e.g., iOSAT, potassium iodide oral solution USP, 65 mg / mL) once daily for 3 days from day 1, starting at least 30 minutes before administration of I-p5+14; 2) Record heart rate, blood pressure, weight, height and concomitant medications; 3) In a cohort of approximately 25 participants, assess the corrected QT (QTc) interval. 124 Obtain a triple 12-lead electrocardiogram (ECG) within 2 hours prior to administering I-p5+14.

[0557] 124 After completion of I-p5+14 administration: 1) A cohort of approximately 25 participants who underwent baseline QTc evaluation will receive a triple 12-lead ECG; 2) Participants will undergo PET / CT scans.

[0558] PET / CT imaging is, 124 This will be performed on the first day of the visit following the administration of I-p5+14. Participants will be: 124 Four hours (±60 minutes) after administration of I-p5+14, a PET / CT scan is performed from the top of the head to the upper thigh. The total PET / CT scan time is estimated to be 45 minutes, including 20 minutes at the cardiac bed position.

[0559] All participants have a baseline 12-lead ECG obtained during screening. For a cohort of approximately 25 participants, 124 Within 2 hours before administering I-p5+14, 124 Immediately after the end of I-p5+14 administration and 124 30 minutes (+10 minutes) after I-p5+14 administration, take a triple ECG to assess the QTc interval.

[0560] Image interpretation Participants are classified into one of the following categories based on (i) visual evaluation of PET / CT images by an independent radiologist and (ii) the status of cardiac amyloid disease based on clinical truth criteria: a) True Positive (TP): Visually positive from PET / CT images 124a) Participants with I-p5+14 cardiac uptake and confirmed positive cardiac amyloidosis according to the true criteria, b) True negative (TN): Visually negative from PET / CT images 124 c) Participants with I-p5+14 cardiac uptake and confirmed negative cardiac amyloidosis by the true criterion, false positive (FP): visually positive from PET / CT images 124 d) Participants with I-p5+14 cardiac uptake and confirmed negative cardiac amyloidosis by the true criterion, false negative (FN): visually negative from PET / CT images 124 Participants with I-p5+14 cardiac uptake and confirmed positive cardiac amyloidosis according to the truth criterion.

[0561] Sensitivity is defined as the number of TP participants divided by the total number of TP and FN participants: Sensitivity = TP / (TP + FN).

[0562] Specificity is defined as the number of TN participants divided by the total number of TN and FP participants: Specificity = TN / (TN + FP).

[0563] endpoint The primary objective of this study is to assess the diagnosis of cardiac amyloidosis. 124 The objective is to evaluate the effectiveness of I-p5+14. The corresponding co-primary endpoint is as follows: 1) Diagnosis of cardiac amyloidosis based on visual scan interpretation. 124 1) Sensitivity of PET / CT imaging using I-p5+14, and 2) Diagnosis of cardiac amyloidosis based on visual scan interpretation. 124 Specificity of PET / CT imaging using I-p5+14. The secondary primary hypotheses are as follows: 1) Null hypothesis: Regarding the diagnosis of cardiac amyloidosis. 124 Sensitivity of imaging using I-p5+14 = 0.65, Alternative hypothesis: Regarding the diagnosis of cardiac amyloidosis. 124 Sensitivity of imaging using I-p5+14 > 0.65, and 2) Null hypothesis: Regarding the diagnosis of cardiac amyloidosis124 Specificity of imaging using I-p5+14 = 0.55, Alternative hypothesis: Regarding the diagnosis of cardiac amyloidosis. 124 The specificity of imaging using I-p5+14 is >0.55. Sensitivity and specificity are calculated for each of the three radiologists. The trial is considered successful if both the sensitivity and specificity null hypotheses are rejected by at least the same two of the three radiologists. Sensitivity and specificity summaries are provided along with two-sided 95% confidence intervals (CIs) using the exact method. For each of sensitivity and specificity, if the aforementioned null hypotheses are rejected by the binomial test, the lower limit of the two-sided 95% CI will mathematically exceed the 65% (sensitivity) or 55% (specificity) threshold for the secondary primary endpoint.

[0564] Subgroup analysis of the primary endpoint will be performed using two-sided 95% confidence intervals (α=0.025) for the following populations: 1) type of systemic amyloidosis (AL, ATTR), 2) sex (female, male), and 3) age (age > 65, age ≤ 65).

[0565] Example 6: Compared with 99mTc-PYP 124 Early cardiac amyloid detection by PET / CT imaging of I-p5+14 In the case of 99mTc-PYP 124 To evaluate the potential consequences of switching between diagnostic methods using PET / CT imaging and those using I-p5+14, 24 individuals who had previously been tested for cardiac amyloidosis were included. 124 I-p5+14 was re-evaluated using PET / CT imaging. 124 PET / CT imaging of I-p5+14 was performed according to the method described in Example 1.

[0566] The results are shown in Table 13 below. Of the seven individuals who tested positive for cardiac amyloid with 99mTc-PYP, all seven tested positive. 124He was diagnosed as positive for cardiac amyloid with I-p5+14. Of the 17 individuals diagnosed as negative for cardiac amyloid with 99mTc-PYP, 11 were 124 I-p5+14 was determined to be positive for cardiac amyloid on PET / CT imaging. These results, compared to 99mTc-PYP, 124 This demonstrates improved sensitivity in detecting cardiac amyloid using I-p5+14. [Table 13]

[0567] Example 7: 124 Diagnostic threshold using I-p5+14 acquisition In 16 individuals without systemic amyloidosis and 27 individuals with systemic amyloidosis (AL, ATTR, or other), organ-specific SUVR was quantified using computer vision analysis of PET / CT images from each individual. 124 Organ-specific uptake of I-p5+14 was calculated. Organ-specific SUVR values ​​were calculated for the liver, spleen, kidneys, pancreas, heart, and adrenal glands.

[0568] To quantify cardiac uptake of radioactive tracers, automated organ segmentation was used. The heart and aorta were automatically segmented on CT data using a 3D pre-trained convolutional neural network. The contours were applied to PET data to quantify mean-normalized uptake values ​​and other uptake parameters in the heart.

[0569] The diagnostic threshold was calculated by determining the 97.5th percentile of the organ-specific SUVR mean value in healthy individuals, relative to the organ-specific SUVR mean value for each organ. For each organ, the diagnostic threshold was calculated by taking the maximum SUVR and the standard deviation of intra-organ uptake (SUVR). hetero ), the coefficient of variation (SUVR heterogeneity) is calculated as the sum of total uptake within the organ normalized by the SUVR peak, the voxel size of the image (SUVR total), and the SUVR mean. covThe same method was used to calculate the value of ).

[0570] If amyloid exceeding the thresholds shown in Table 14 was detected in an organ, the individual was diagnosed with systemic amyloidosis. [Table 14]

Claims

1. A method for diagnosing early-stage amyloid-related disease in individuals at risk of developing amyloid-related disease, wherein the method is: a. Administering an individual an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14, conjugated to a detectable label, and b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual. The method comprising, where the amount of amyloid-reactive peptide exceeding a threshold indicates that the individual has an amyloid-related disease, and the amyloid-related disease is an early stage of amyloid-related disease.

2. The method according to claim 1, for determining that an individual is at risk of amyloid-related disease based on the presence of a gene mutation, having multiple myeloma, having amyloid-positive laminectomy tissue, having amyloid-positive tissue derived from carpal tunnel release surgery, having monoclonal immunoglobulinemia of unknown significance (MGUS), having left ventricular impulse-preserving heart failure (HFpEF), having left ventricular impulse-reducing heart failure (HFrEF), being from a susceptible ethnic group, or being elderly.

3. The method according to claim 2, wherein the gene mutation is located within the transthyretin protein.

4. The method according to claim 2, wherein the gene mutation is located within the fibrinogen α protein.

5. The method according to any one of claims 1 to 3, wherein the individual does not exhibit symptoms of amyloidosis.

6. The method according to any one of claims 1 to 3, wherein the individual has neurological symptoms of amyloid disease.

7. The method according to any one of claims 1 to 6, wherein the individual has been previously diagnosed as not having amyloid-related disease.

8. A method for diagnosing early-stage amyloid-related disease in an individual suspected of having amyloid-related disease, the method being: a. Administering an individual an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14, conjugated to a detectable label, and b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual. The method comprising, where the amount of amyloid-reactive peptide exceeding a threshold indicates that the individual has an amyloid-related disease, and the amyloid-related disease is an early stage of amyloid-related disease.

9. a. Quantifying amyloid in an individual by administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14, conjugated to a detectable label, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs. b. Administer an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs: 1 to 14, conjugated to a detectable label, to an individual, and quantify amyloid in the individual at a second time point by detecting the amount of the detectable label in the individual's tissues or organs, and c. Determining the prognosis of an individual by comparing the amyloid quantified at a first time point in the tissue or organ with the amyloid quantified at a second time point in the same tissue or organ. A method for determining the prognosis of individuals with amyloid-related diseases, including [specific example].

10. The method according to claim 8, wherein the prognosis of an individual with amyloid-related disease is based on detecting the amount of amyloid-reactive peptide in the heart only.

11. The method according to claim 8, wherein the prognosis of an individual with amyloid-related disease is based on detecting the amount of amyloid-reactive peptide in the heart and kidneys.

12. The method according to claim 8, wherein the prognosis of an individual with amyloid-related disease is based on detecting the amount of amyloid-reactive peptide in the heart and kidneys and all other organs.

13. a. Administering an amyloid-reactive peptide containing the amino acid sequence described in any one of SEQ ID NOs: 1 to 14, conjugated to a detectable label, b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual, and c. Initiating treatment for amyloid-related disease when the amount of amyloid-reactive peptide exceeds a threshold. Treatment methods for amyloid-related diseases, including those mentioned above.

14. The method according to claim 13, wherein the individual has an early-stage amyloid-related disease.

15. a. Quantifying amyloid in an individual by administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14, conjugated to a detectable label, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs. b. To provide treatment for amyloid-related diseases. c. Administer an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14, conjugated to a detectable label, to an individual, and quantify amyloid in the individual at a second time point by detecting the amount of the detectable label in the individual's tissues or organs, and d. To determine whether the treatment is effective by comparing the amyloid quantified at the first time point in the tissue or organ with the amyloid quantified at the second time point in the same tissue or organ. A method for monitoring the treatment of amyloid-related diseases in an individual, including [specific example].

16. The method according to any one of claims 1 to 15, further comprising administering an amyloid-reactive peptide containing an amino acid sequence described in any one of Sequence IDs 1 to 14 conjugated to a detectable label to an individual, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs, thereby quantifying amyloid in the individual at a second time point, optionally a third time point, a fourth time point, and / or a fifth time point.

17. The method according to claim 15 or 16, wherein the first time point and the second time point are separated by at least six weeks.

18. A method for selecting a treatment for amyloid-related disease in an individual, the method being: a. Administering an individual an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14, conjugated to a detectable label, and b. Detecting the amount of amyloid-reactive peptide by detecting the amount of detectable label in the tissues or organs of an individual. The method comprising, wherein if amyloid is detected in the heart, treatment for amyloid-related disease is performed, and if amyloid is not detected in the heart, alternative therapy is performed.

19. a. Quantifying amyloid in an individual by administering an amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14, conjugated to a detectable label, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs. b. To provide treatment for amyloid-related diseases. c. An amyloid-reactive peptide containing an amino acid sequence described in any one of SEQ ID NOs: 1 to 14, conjugated to a detectable label, is administered to an individual, and the amount of the detectable label in the individual's tissues or organs is detected to quantify the amyloid in the individual at a second time point. d. Comparing amyloid quantified at a first time point in a tissue or organ with amyloid quantified at a second time point in the same tissue or organ, and e. Adjusting treatment for amyloid-related diseases. A method for managing the treatment of amyloid-related diseases in an individual, including the treatment of amyloid-related diseases in an individual.

20. The method according to claim 19, wherein if the amyloid disease is stable between the first and second time points, the treatment is modified.

21. The method according to claim 19, wherein treatment is resumed if the amyloid load is increasing.

22. The method according to any one of claims 9 to 12 and 14 to 21, wherein the amount of amyloid-reactive peptide exceeding a threshold indicates that an individual has an amyloid-related disease.

23. The method according to claim 22, wherein the amyloid-related disease is an early stage of amyloid-related disease.

24. The method according to any one of claims 1 to 23, wherein the amyloid-reactive peptide comprises SEQ ID NO:

13.

25. The method according to any one of claims 1 to 24, wherein the detectable label is a radioactive label.

26. The detectable label is １１ C, １８ F, １１１ In, ９９ｍ Tc, ８９ Zr, ６８ Ga and １２３ I, １２４ I and １２５ I, selected from the group consisting of, the method according to any one of claims 1 to 25.

27. Detectable signs １２４ The method according to any one of claims 1 to 26, wherein I.

28. Doses of approximately 0.1 to approximately 25 mCi １２４ The method according to any one of claims 1 to 27, comprising administering an I-labeled amyloid-reactive peptide to an individual.

29. Doses of approximately 0.3 mCi, 1 mCi, or 2 mCi １２４ The method according to any one of claims 1 to 28, comprising administering an I-labeled amyloid-reactive peptide to an individual.

30. A dose of approximately 1 mCi (±10%) １２４ The method according to any one of claims 1 to 29, comprising administering an I-labeled amyloid-reactive peptide to an individual.

31. The method according to any one of claims 1 to 30, comprising administering a dose of Tc-99 of approximately 20 mCi to an individual.

32. The method according to any one of claims 1 to 31, comprising administering to an individual about 0.3 mg, 1 mg, 1.5 mg, or about 2 mg of an amyloid-reactive peptide.

33. The method according to any one of claims 1 to 32, comprising administering an amyloid-reactive peptide intravenously.

34. The method according to any one of claims 1 to 33, further comprising determining organ-specific standard uptake ratios for organs within an individual.

35. The method according to claim 34, wherein the organ-specific SUVR for an individual is selected from the group consisting of the mean SUVR, the maximum SUVR, and the peak SUVR.

36. The method according to any one of claims 1 to 35, further comprising determining organ-specific SUVs for an individual.

37. The method according to claim 36, wherein the organ-specific SUV for an individual is selected from the group consisting of SUV mean, SUV maximum, and SUV peak.

38. The method according to any one of claims 1 to 37, further comprising determining an organ-specific dose per gram percentage (%ID) for an individual.

39. The method according to any one of claims 1 to 38, wherein the sensitivity for detecting amyloid using an amyloid-reactive peptide is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%.

40. The method according to any one of claims 1 to 39, wherein the sensitivity is approximately 99% or approximately 100%.

41. The method according to any one of claims 1 to 40, wherein the specificity of the amyloid-reactive peptide is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100%.

42. The method according to any one of claims 1 to 41, wherein the specificity of the amyloid-reactive peptide is about 99% or 100%.

43. The method according to any one of claims 1 to 42, further comprising determining one or more health-related quality of life indicators for an individual.

44. The method according to any one of claims 1 to 43, further comprising detecting one or more biomarkers associated with amyloid-related disease.

45. The method according to claim 44, wherein a biomarker associated with amyloid-related disease is selected from the group consisting of troponin T, NTproBNP, urinary protein levels, UACR, EGFR, and alkaline phosphatase levels.

46. The method according to any one of claims 1 to 45, wherein the tissue or organ of the individual is selected from the group consisting of lungs, fat, heart, kidneys, pancreas, joints, spine, liver, spleen, adrenal gland, bone lesions, choroid plexus, pituitary gland, uterus, bone marrow, musculoskeletal tissue, gastrointestinal tract, and prostate.

47. The method according to any one of claims 1 to 8, 13, and 22 to 46, wherein the threshold is the SUVR average threshold.

48. If the organ or tissue is the liver, the mean SUVR threshold is approximately 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, or 1.6; if the organ or tissue is the spleen, the mean SUVR threshold is approximately 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5; if the organ or tissue is the kidney, the mean SUVR threshold is approximately 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, or 2.3; and if the organ or tissue is the pancreas, The method according to claim 47, wherein the average SUVR threshold is approximately 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, or 1.7, and if the organ or tissue is the heart, the average SUVR threshold is approximately 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, or 2.1, or if the organ or tissue is the adrenal gland, the average SUVR threshold is approximately 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, or 1.

4.

49. The method according to claim 48, wherein the mean SUVR threshold is 1.31 when the organ or tissue is the liver, 1.21 when the organ or tissue is the spleen, 1.88 when the organ or tissue is the kidney, 1.40 when the organ or tissue is the pancreas, 1.19 when the organ or tissue is the heart, or 1.35 when the organ or tissue is the adrenal gland.

50. The method according to any one of claims 1 to 8, 13, and 22 to 46, wherein the threshold is the SUVR maximum threshold.

51. If the organ or tissue is the liver, the SUVR maximum threshold is 10.6, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, or 16; if the organ or tissue is the spleen, the SUVR maximum threshold is 3.8, 4, 4.5, 5, 5.5, or 5.9; and if the organ or tissue is the kidney, the SUVR maximum threshold is 7.7, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, or 11. The method according to claim 50, wherein the maximum SUVR threshold is 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11 or 11.4 if the organ or tissue is the pancreas, or 4.9, 5, 5.5, 6, 6.5, 7 or 7.5 if the organ or tissue is the heart, or 1, 1.1, 1.2, 1.3, 1.4, 1.5 or 1.6 if the organ or tissue is the adrenal gland.

52. The method according to claim 51, wherein the maximum SUVR threshold is 13.26 when the organ or tissue is the liver, 4.86 when the organ or tissue is the spleen, 9.68 when the organ or tissue is the kidney, 9.48 when the organ or tissue is the pancreas, 6.20 when the organ or tissue is the heart, or 1.31 when the organ or tissue is the adrenal gland.

53. The method according to any one of claims 1 to 52, further comprising performing a cardiac biopsy if amyloid is detected in the heart.

54. The method according to any one of claims 1 to 53, further comprising performing additional amyloid imaging on the individual.

55. The method according to any one of claims 1 to 54, further comprising performing additional amyloid imaging on the individual before administering the amyloid-reactive peptide.

56. The method according to claim 55, wherein additional amyloid imaging of the individual includes ECHO, CMR, bone scintigraphy, or positron emission tomography imaging.

57. Additional amyloid imaging of the individual, ９９ｍ Tc-PyP, ９９ｍ Tc-DPD, ９９ｍ Tc-HMDP, ９９ｍ Tc-MDP, other bone scintigraphy tracers, １２４ I and １８ F-Florbetapyr, １８ F-Frutemethamol and １８ The method according to claim 56, further comprising a tracer selected from the group consisting of F-florbetabene.

58. The method according to any one of claims 1 to 57, wherein the diagnosis, prognosis, or response is confirmed by an indicator of the individual's health-related quality of life, the presence or amount of one or more biomarkers associated with amyloid-related disease, cardiac biopsy, and / or additional amyloid imaging of the individual.

59. A method according to any one of claims 1 to 58, wherein an individual is diagnosed with amyloid cardiomyopathy.

60. The method according to any one of claims 1 to 59, wherein the individual is suspected of having amyloid cardiomyopathy.

61. The method according to any one of claims 1 to 60, wherein the amyloid-related disease is systemic or localized amyloidosis.

62. The method according to any one of claims 1 to 61, wherein the amyloid-related disease is cardiac amyloidosis.

63. The method according to any one of claims 1 to 62, wherein the amyloid-reactive peptide has pan-amyloid specificity.

64. Amyloid-reactive peptides include immunoglobulin light chain (AL), immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin (ATTR wild type; ATTR mutant), apolipoprotein AI (AApoAI), apolipoprotein AII (AApoAII), apolipoprotein AIV (AApoAIV), gelzolin (AGel), apolipoprotein C-II (AApoCII), apolipoprotein C-III (AApoCIII), lysozyme (ALys), and leukocytes. The method according to any one of claims 1 to 63, comprising binding to a chemotactic factor (ALECT2), fibrinogen α variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP); α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), IAAP, ALβ4, or ALβ1 amyloid.

65. The method according to any one of claims 1 to 64, wherein amyloid-reactive peptides are detected using PET, PET / CT, gamma machinography, SPECT, and / or SPECT / CT.

66. The method according to any one of claims 1 to 65, wherein the amyloid-related disease is selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AApoCII, AApoCIII, AGel, ALys, ALECT2, AFib, ACYs, ACaal, AMed, AIAPP, APro, AIns, APrP, and Aβ amyloidosis.

67. The method according to claim 66, wherein the amyloid-related disease is cardiac AL amyloidosis.

68. The method according to claim 66, wherein the amyloid-related disease is cardiac ATTR amyloidosis.

69. The method according to any one of claims 1 to 68, wherein the individual has a genetic predisposition to amyloid-related disease.

70. The method according to any one of claims 1 to 69, wherein the individual has a family history of amyloid-related disease.

71. The method according to any one of claims 1 to 70, wherein the individual is elderly.

72. The method according to any one of claims 1 to 71, wherein the individual has an early-stage amyloid-related disease.

73. The method according to any one of claims 1 to 72, wherein the individual has early-stage systemic amyloidosis.

74. The method according to any one of claims 1 to 73, wherein the individual has early-stage cardiac amyloidosis.

75. The method according to any one of claims 1 to 74, wherein the individual has early-stage AL amyloidosis.

76. The method according to claim 75, wherein early-stage AL amyloidosis is diagnosed according to the Mayo Clinic system.

77. The method according to claim 76, wherein the early stage of AL amyloidosis is stage 1 AL amyloidosis.

78. The method according to any one of claims 1 to 77, wherein the individual has early-stage ATTR amyloidosis.

79. The method according to claim 78, wherein early-stage ATTR amyloidosis includes stage 1 ATTR amyloidosis.

80. The method according to any one of claims 1 to 12, 16 to 18, and 20 to 79, further comprising providing treatment for amyloid-related disease.

81. The method according to any one of claims 13 to 15, 19 and 80, wherein the treatment for amyloid-related disease is selected from the group consisting of transthyretin stabilizers (e.g., tafamidis, acolamidis), transthyretin silencers (e.g., patisirane, inotercene, butricirane, eprotercen), gene editing approaches, anti-amyloid approaches utilizing monoclonal antibodies, therapies targeting plasma cell clones (e.g., daratumumab, bortezomib), and antibody-peptide fusions comprising an antibody-peptide fusion protein containing a second amyloid-reactive peptide and an antibody that binds to amyloid fibril.

82. The method according to claim 81, wherein the second amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 14, comprising 0, 1, 2, 3, or 4 amino acid substitutions, insertions, or deletions.

83. The method according to claim 81 or 82, wherein the antibody-peptide fusion protein comprises a heavy chain and a light chain, and an amyloid-reactive peptide is linked to the C-terminus of the antibody's light chain via a spacer.

84. Antibody-peptide fusion proteins, (i) an amyloid-reactive peptide containing the amino acid sequence described in Sequence ID No. 2, and (ii) Antibodies that bind to human amyloid fibrils The method according to any one of claims 81 to 83, wherein the antibody comprises a heavy chain and a light chain, the heavy chain of the antibody comprises a heavy chain variable region (VH), and the light chain of the antibody comprises a light chain variable region (VL), wherein VH comprises CDR-H1 comprising the amino acid sequence described in SEQ ID NO: 106, CDR-H2 comprising the amino acid sequence described in SEQ ID NO: 107, and CDR-H3 comprising the amino acid sequence described in SEQ ID NO: 108, and VL comprises CDR-L1 comprising the amino acid sequence described in SEQ ID NO: 103, CDR-L2 comprising the amino acid sequence described in SEQ ID NO: 104, and CDR-L3 comprising the amino acid sequence described in SEQ ID NO: 105, wherein the amyloid-reactive peptide and the antibody are linked at the C-terminus of the light chain, and the amyloid-reactive peptide is linked to the antibody via a spacer comprising the amino acid sequence described in SEQ ID NO:

83.

85. The method according to any one of claims 1 to 84, comprising administering a peptide containing an amyloid-reactive peptide and an N-terminal leader sequence to an individual.

86. The method according to claim 85, wherein the peptide containing the amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO:

100.

87. The method according to claim 86, wherein a detectable label is bound to one or more amino acids at positions 1 to 6 of the amino acid sequence described in Sequence ID No.

100.

88. Detectable signs １２４ The method according to claim 87, wherein I.

89. １２４ The method according to claim 88, wherein I is bonded to amino acid 4 of SEQ ID NO:

100.

90. The method according to any one of claims 1 to 89, further comprising administering potassium iodide to an individual.

91. The method according to claim 90, wherein the administration of potassium iodide comprises administering 130 mg of potassium iodide to the patient for 7 days, starting one day before the administration of the amyloid-reactive peptide.

92. The method according to claim 91, wherein the administration of potassium iodide comprises administering 130 mg of potassium iodide to the patient for three days, starting approximately 30 minutes before the administration of the amyloid-reactive peptide.

93. The method according to any one of claims 90 to 92, wherein the administration of potassium iodide is by oral administration.

94. A peptide comprising the amino acids described in Sequence ID No. 100, wherein the peptide includes a detectable label bound to one or more amino acids at positions 1 to 6.

95. Detectable signs １２４ The peptide according to claim 94, wherein I.

96. １２４ The peptide according to claim 95, wherein I is bound to amino acid 4 of SEQ ID NO: 100.

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