Methods for diagnosing amyloidosis

Abstract

The present disclosure relates in some aspects to methods and compositions for detecting amyloid-related diseases in an individual.

Description

Cross-reference to related applications

[0001] This application claims priority to U.S. Provisional Application No. 62 / 581,213, filed September 7, 2023, and U.S. Provisional Application No. 63 / 657,715, filed June 7, 2024, the contents of which are incorporated herein by reference in their entirety. Reference to electronic sequence listing

[0002] The contents of the electronic sequence list (165992001240SEQLIST.xml; size: 40,979 bytes; creation date: September 6, 2024) are incorporated herein by reference in their entirety. Technical Field

[0003] This disclosure relates in some respects to methods for diagnosing amyloid-related diseases. Background of the Invention

[0004] Systemic amyloidosis (SAM) is a progressive protein misfolding disorder characterized by the extracellular deposition of proteinaceous fibrils, extracellular matrix components, and serum proteins. The accumulation of amyloid proteins, particularly in the heart and kidneys (the organs most affected in all types of amyloidosis), leads to organ dysfunction, poor quality of life, and ultimately death. Approximately 20 different proteins have been identified as fibril components in SAM (AimoA, Merlo M, Porcari A, ...). et al. Eur J Heart Fail. 2022); however, patients with variant or wild-type transthyretin- (ATTRv and ATTRwt), immunoglobulin light chain- (AL), and leukocyte chemokine 2- (ALECT2)-associated amyloidosis account for more than 90% of cases diagnosed in the United States (Edwards CV, Rao N, Bhutani D, 2022). et al. Blood. 2021;138:2632–2641).

[0005] Cardiac amyloidosis is the more severe manifestation of amyloidosis associated with AL and ATTR, causing restrictive hypertrophic cardiomyopathy, particularly affecting the left ventricular wall and interventricular septum, with 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 may be involved, leading to heterogeneous clinical presentations and making accurate and rapid diagnosis challenging (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, ParkM-A, Cuddy S, et al. J Nucl Med. 2021;62:716–722). Echocardiography and cardiac magnetic resonance imaging (CMR) can reveal structural abnormalities and functional sequelae associated with amyloidosis10; however, these techniques are not specific to amyloid protein. 99m Tc-pyrophosphate ( 99m Tc-PyP) or 99m Nuclear imaging of Tc-3,3-bisphosphono-1,2-propanedicarboxylic acid (DPD) is commonly used to diagnose cardiac ATTR-associated amyloidosis. These bone-seeking agents bind to cardiac microcalcifications associated with cardiac amyloid and preferentially detect only ATTR-associated cardiac amyloidosis (Antoni G, Lubberink M, Estrada S, et al. J Nucl Med. (2013;54:213–220). In some respects, existing methods are limited by the unavailability of amyloid-specific measures for monitoring changes in amyloid load over time. Therefore, non-invasive and quantitative measures for detecting and monitoring cardiac and extracardiac amyloid deposits are clinically beneficial. This article provides methods and compositions for addressing these and other needs. Invention Overview

[0007] In one aspect, this article provides a method for early diagnosis of amyloid-related disease in an individual at risk of developing the disease, comprising a) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs, wherein an amount of the amyloid-reactive peptide above a threshold indicates that the individual has amyloid-related disease, wherein the amyloid-related disease is in its early stages.

[0008] In some embodiments, an individual is identified as being at risk for amyloid-related diseases based on the presence of a gene mutation, having multiple myeloma, having amyloid-positive laminectomy tissue, having amyloid-positive tissue from carpal tunnel release surgery, having monoclonal gammopathy of undetermined 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 older. In some embodiments, the gene mutation is in transthyretin. In some embodiments, the gene mutation is in fibrinogen α protein.

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

[0010] In another aspect, this article provides a method for early diagnosis of amyloid-related disease in an individual suspected of having it, comprising a) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs, wherein an amount of the amyloid-reactive peptide above a threshold indicates that the individual has amyloid-related disease, wherein the amyloid-related disease is in its early stages.

[0011] In another aspect, this article provides a method for determining the prognosis of an individual suffering from early-stage amyloid-associated disease, comprising: a) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated with any one of SEQ ID NO: 1-14, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs to first quantify amyloid in the individual; b) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated with any one of SEQ ID NO: 1-14, and detecting the amount of the detectable marker in the individual's tissues or organs to second quantify amyloid in the individual; and c) comparing the first quantification of amyloid in the tissues or organs with the second quantification of amyloid in the tissues or organs to determine the prognosis of the individual, wherein the amyloid-associated disease is in its early stage.

[0012] In some embodiments, the prognosis of an individual with amyloid-associated disease is based on the measurement of amyloid-reactive peptides in the heart alone. In some embodiments, the prognosis of an individual with amyloid-associated disease is based on the measurement of amyloid-reactive peptides in the heart and kidneys. In some embodiments, the prognosis of an individual with amyloid-associated disease is based on the measurement of amyloid-reactive peptides in the heart, kidneys, and all other organs.

[0013] In another aspect, this article provides a method for treating amyloid-associated disease in an individual suffering from early-stage amyloid-associated disease, comprising a) administering an amyloid-reactive peptide to the individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; b) detecting the amount of amyloid-reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs; and c) if the amount of amyloid-reactive peptide is above a threshold, administering treatment for amyloid-associated disease.

[0014] In some implementations, the individual is suspected of having amyloid-related disease.

[0015] In another aspect, this article provides a method for monitoring treatment of amyloid-associated disease in an individual suffering from an early stage of amyloid-associated disease, comprising a) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated with any one of SEQ ID NO: 1-14, 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 first quantify amyloid in the individual; b) administering treatment for amyloid-associated disease; c) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated with any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to second quantify amyloid in the individual; and d) comparing the amyloid in the tissues or organs first quantified with the amyloid in the tissues or organs second quantified to determine whether the treatment is effective.

[0016] In some embodiments, the method further includes administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs, to quantify amyloid in the individual a second, optionally a third, fourth, and / or fifth time.

[0017] In some implementations, the first and second times are spaced at least 6 weeks apart.

[0018] In another aspect, this article provides a method for selecting a treatment for an amyloid-associated disease in an individual, comprising a) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs, wherein if amyloid is detected in the heart, treatment for the amyloid-associated disease is administered, and wherein if amyloid is not detected in the heart, an alternative therapy is administered.

[0019] In another aspect, this document provides a method for managing the treatment of amyloid-related diseases in an individual, comprising: a) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated with any one of SEQ ID NO: 1-14, 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 first quantify amyloid in the individual; b) administering the treatment for amyloid-related diseases; c) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated with any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to second quantify amyloid in the individual; and d) comparing the amyloid in the tissues or organs first quantified with the amyloid in the tissues or organs second quantified; and e) adjusting the treatment for amyloid-related diseases.

[0020] In some implementations, if amyloidosis is stable between the first and second treatments, the treatment is changed. In some implementations, if the amyloid burden increases, treatment is restarted.

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

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

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

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

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

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

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

[0028] In some embodiments, the method further includes determining the organ-specific standard uptake ratio (SUVR) for an individual's internal organs. In some embodiments, the organ-specific SUVR for an individual is selected from a group consisting of the mean SUVR, the maximum SUVR, and the peak SUVR.

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

[0030] In some embodiments, the method further includes determining the organ-specific percentage injection dose / gram (%ID) for an individual.

[0031] In some embodiments, the sensitivity of using amyloid reactive peptides to detect amyloid 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 about 99% or 100%.

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

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

[0035] In some implementations, the individual's tissues or organs are selected from the group consisting of the 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.

[0036] In some implementations, the threshold is the average SUVR threshold. In some implementations, if the organ or tissue is the liver, the average 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 average 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 average SUVR threshold is approximately 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, or 2.3. If the organ or tissue is the pancreas, the average 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 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. In some implementations, the average SUVR threshold is 1.31 if the organ or tissue is the liver, 1.21 if the organ or tissue is the spleen, 1.88 if the organ or tissue is the kidney, 1.40 if the organ or tissue is the pancreas, 1.19 if the organ or tissue is the heart, or 1.35 if the organ or tissue is the adrenal gland.

[0037] In some implementations, the threshold is the maximum SUVR threshold. In some implementations, 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, or 10. The maximum SUVR thresholds are 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. In some implementations, the maximum SUVR threshold is 13.26 if the organ or tissue is the liver, 4.86 if the organ or tissue is the spleen, 9.68 if the organ or tissue is the kidney, 9.48 if the organ or tissue is the pancreas, 6.20 if the organ or tissue is the heart, or 1.31 if the organ or tissue is the adrenal gland.

[0038] In some embodiments, the method further includes performing a cardiac biopsy if amyloid is detected in the heart. In some embodiments, the method further includes performing additional amyloid imaging of the individual. In some embodiments, the method further includes performing additional amyloid imaging of the individual prior to the administration of an amyloid-reactive peptide. In some embodiments, additional amyloid imaging of the individual includes ECHO, CMR, bone scintillation, or positron emission tomography. In some embodiments, additional amyloid imaging of the individual further includes a tracer selected from the group consisting of: 99m Tc-PyP, 99m Tc-DPD, 99m Tc-HMDP, 99m Other bone scintillation tracers for Tc-MDP 124 I and 18 F-Fluorobetapir 18 F-Flutemetamol and 18 F-fluorobetabenzene.

[0039] In some implementations, diagnosis, prognosis, or response is confirmed by measures of an 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 implementations, the individual is diagnosed with amyloid cardiomyopathy. In some implementations, the individual is suspected of having amyloid cardiomyopathy.

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

[0042] In some embodiments, the amyloid-reactive peptide is pan-amyloid-specific. In some embodiments, the amyloid-reactive peptide binds to the following amyloid proteins: 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 (AGel), apolipoprotein C-II (AApoCII), apolipoprotein C-II (AApoCIII), lysozyme (ALys), leukocyte chemokine (ALECT2), fibrinogen a variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactodherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), and prion protein (priority). protein (APrP); α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), IAAP, ALβ4 or ALβ1.

[0043] In some implementations, PET, PET / CT, gamma scintillation scanning, SPECT, and / or SPECT / CT are used to detect amyloid reactive peptides.

[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, ACal, AMed, AIAPP, APro, AIns, APrP, and Aβ amyloidosis. In some embodiments, amyloid-related disease is cardiac AL amyloidosis. In some embodiments, amyloid-related disease is cardiac ATTR amyloidosis.

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

[0046] In some implementations, the individual is an elderly person.

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

[0048] In some embodiments, the method further includes administering treatment for amyloid-related diseases. In some embodiments, the treatment for amyloid-related diseases is selected from the group consisting of: transthyretin stabilizers (e.g., tafamidis, acoramidis), transthyretin silencers (e.g., Patisiran, Inotersen, Vutrisiran, Eploetersen), gene editing methods, anti-amyloid methods using monoclonal antibodies, treatments targeting plasma cell clones (e.g., daratumumab, bortezomib), and antibody-peptide fusions comprising a second amyloid reactive peptide and an antibody binding to amyloid fibrils. In some embodiments, the second amyloid reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-14, comprising 0, 1, 2, 3, or 4 amino acid substitutions, insertions, or deletions. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain and a light chain, wherein the amyloid-reactive peptide is linked to the C-terminus of the light chain of the antibody via a spacer. In some embodiments, the antibody-peptide fusion protein comprises: (i) an amyloid reactive peptide comprising the amino acid sequence shown in SEQ ID NO: 2; and (ii) an antibody that binds to human amyloid fibrils, 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 the VH comprises: CDR-H1 comprising the amino acid sequence shown in SEQ ID NO: 106, CDR-H2 comprising the amino acid sequence shown in SEQ ID NO: 107, and CDR-H3 comprising the amino acid sequence shown in SEQ ID NO: 108, and the VL comprises: CDR-L1 comprising the amino acid sequence shown in SEQ ID NO: 103, CDR-L2 comprising the amino acid sequence shown in SEQ ID NO: 104, and CDR-L3 comprising the amino acid sequence shown in SEQ ID NO: 105; wherein the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain, and wherein the amyloid reactive peptide is linked to the antibody via a spacer comprising the amino acid sequence shown in SEQ ID NO: 83.

[0049] In some embodiments, a peptide comprising an amyloid-reactive peptide and an N-terminal leader sequence is administered to an individual. In some embodiments, the peptide comprising the amyloid-reactive peptide comprises the amino acid sequence shown in SEQ ID N: 100.

[0050] In some embodiments, the detectable marker binds to one or more amino acids at positions 1-6 of the amino acid sequence shown in SEQ ID NO: 100. In some embodiments, the detectable marker is... 124 I. In some implementations, 124 I binds to amino acid 4 of SEQ ID NO: 100.

[0051] In some embodiments, the method further includes administering potassium iodide to the individual. In some embodiments, administering potassium iodide comprises administering 130 mg of potassium iodide to the patient for seven days, starting one day before administering the amyloid-reactive peptide. In some embodiments, administering potassium iodide comprises administering 130 mg of potassium iodide to the patient for three days, starting approximately 30 minutes before administering the amyloid-reactive peptide. In some embodiments, the potassium iodide is administered orally.

[0052] In another aspect, this document provides peptides comprising the amino acids shown in SEQ ID NO: 100, wherein the peptide contains a detectable tag binding to one or more amino acids at positions 1-6. In some embodiments, the detectable tag is 124 I. In some implementations, 124 I binds to amino acid 4 of SEQ ID NO: 100. Brief description of the attached diagram

[0053] The accompanying drawings illustrate certain features and advantages of this disclosure. These embodiments are not intended to limit the scope of the appended claims in any way.

[0054] Figure 1 This study describes a multipart, open-label study to evaluate [the patient's condition] in patients with any type of systemic amyloidosis; presymptomatic subjects carrying germline TTR mutations; and healthy subjects. 124 Safety, dosing assays and biodistribution of I-P5+14 peptide.

[0055] Figure 2 depicts the biodistribution of radioactivity in healthy subjects. Figure 2A The maximum intensity projection is shown for PET data obtained 5 hours after infusion from five healthy subjects. Figure 2B The image shows a transaxial PET / CT image of the heart in a healthy subject.

[0056] Figure 3 depicts patients with AL and ATTR-associated amyloidosis. 124 Cardiac uptake of I-p5+14. Figure 3A Displays the maximum intensity projection of cardiac PET data and coronary PET / CT images (image intensity is set individually for optimal visualization). Figure 3BThis image shows transverse PET / CT images of the heart in healthy subjects and patients with AL or ATTR amyloidosis who are considered positive (Clin+) or negative (Clin-) for cardiac amyloidosis in their medical records and have positive PET imaging (PET+).

[0057] Figure 4 depicts the heart. 124 I-P5+14SUVR 平均 PET evaluation and its correlation with serum NTproBNP. Figure 4A The cardiac SUVR was shown in patients with AL, ATTR, other types of amyloidosis, and healthy subjects. 平均 The comparison also included cardiac SUVR in PET-positive and PET-negative AL and ATTR patients. 平均 value. Figure 4B-4C The cardiac SUVR was described in AL patients. 平均 A significant positive correlation with serum NTproBNP ( Figure 4B This was not significant in ATTR patients. Figure 4C ). Figure 4D The cardiac SUVR for each patient was shown as scaled by serum creatinine. 平均 Correlation with serum NTproBNP in AL patients.

[0058] Figures 5A-5B The description depicts amyloidosis in patients with AL or ATTR-associated amyloidosis. 124 Extracardiac uptake of I-p5+14. Lungs, fat, kidneys, pancreas, joints, and spine of ATTR amyloidosis patients ( Figure 5A ) and liver, spleen, kidneys, adrenal glands, bone lesions, choroid plexus, pituitary gland, bone marrow, and pancreas in patients with AL amyloidosis and those with multiple myeloma and amyloidosis ( Figure 5B )middle 124 Intake of I-P5+14.

[0059] Figure 6 The cardiac uptake of a radioactive tracer for quantitative analysis was depicted. 124 Automatic iso-contouring on I-P5+14 PET images. LA: Left atrium. LV: Left ventricle. PET / CT: Positron emission tomography / computed tomography. RV: Right ventricle. SD: Standard deviation. VOI: Target volume. %ID / ml: Percentage injection dose in milliliters.

[0060] Figure 7 It describes the use of equivocal.99m In patients with ATTRwt cardiomyopathy (CMP) undergoing PYP SPECT / CT, the intake of... 124 I-P5+14 PET / CT. The individual presented on echocardiography with heart failure, preserved left ventricular ejection fraction, grade 2 diastolic dysfunction, asymmetric hypertrophy, and impaired overall longitudinal strain. He had a history of surgery for bilateral carpal tunnel syndrome and lumbar spinal stenosis, and there was no evidence of abnormal immunoglobulin light chains. 99m Tc-PyP SPECT / CT was inconclusive (non-diagnostic) and showed grade 1 uptake. Subsequently, cardiac MRI showed diffuse late gadolinium enhancement. ATTRwt-CMP (stage 2 NAC, stage 2 Mayo) was confirmed by endocardial myocardial biopsy with mass spectrometry and normal TTR gene sequencing. The procedure was performed within 6 months. 124 I-P5+14 PET / CT showed clear myocardial uptake. NAC: National Center for Amyloidosis. MRI: Magnetic Resonance Imaging. 99m Tc-PYPSPECT / CT: 99m Tc-pyrophosphate single-photon emission computed tomography / computed tomography.

[0061] Figure 8 Depicting 124 I-p5+14 intake metrics. 124 I-p5+14 LV %ID, CAA, and TBR completely distinguished amyloid CMP cases from controls, but SUV 平均 No. The uptake measurement and calculations are explained in the Methods section. The Youden index was used to define the threshold for high indicators. Dunn's test was used for intergroup comparisons. CAA: Cardiac amyloid activity (SUV units × mL). LV: Left ventricle. SUV: Standardized uptake value. TBR: Target to background ratio. %ID: Percentage injection dose.

[0062] Figure 9 Depicting 18 F-Fluorbetapyr uptake index. 18 F-Fluorbetapyr LV %ID and CAA completely distinguished amyloid CMP cases from controls, but SUV 平均 No TBR. The Youden index was used to define the threshold for high indices. The Dunn test was used for inter-group comparisons.

[0063] Figure 10 Describing the same participants 124 I-p5+14 and 18 F-flubetapyr uptake index. Obtained in the same participants. 124 I-p5+14 and18 F-fluorobetapyr imaging (excluding 1 patient without) 18 AL-CMP participants in F-flubetapyr PET / CT). LV %ID, SUV 平均 and CAA in adoption 124 The I-p5+14 is higher in ATTRwt-CMP. However, LV TBR uses 18 F-Fluorobetapyr was higher in AL-CMP and ATTRwt-CMP. Absorption measurements and calculations are explained in the Methods section. P-values ​​were obtained from paired Wilcoxon signed-rank tests.

[0064] Figure 11A-11B Depicting the SUVR 平均 And NTproBNP measured for patients with amyloidosis AL ( Figure 11A ) and ATTR patients ( Figure 11B )of 124 Changes in I-P5+14 uptake.

[0065] Figure 12 The relationship between the percentage change in NTproBNP and the proportional percentage change in cardiac SUVR was depicted in 19 patients with amyloidosis of AL or ATTR.

[0066] Figure 13A-13I The description covers the effects of SUVR on patients with AL, ATTR, and ALL amyloidosis. 平均 Measurements of the heart 124 The relationship between anatomical and functional parameters of I-P5+14 uptake. In AL patients, left ventricular wall thickness (LV) (Y-axis) and cardiac SUVR. 平均 (X-axis) related ( Figure 13A In AL patients, parameters reflecting left ventricular function—global longitudinal strain (GLS) (Y-axis) and cardiac SUVR—are related. 平均 (X-axis) related ( Figure 13B In AL patients, interventricular septal thickness (IVS) (Y-axis) is related to cardiac SUVR. 平均 (X-axis) uncorrelated ( Figure 13C In patients with ATTR, cardiac LV wall thickness (Y-axis) and cardiac SUVR 平均 (X-axis) related ( Figure 13D In ATTR patients, GLS (Y-axis) and cardiac SUVR 平均 (X-axis) uncorrelated ( Figure 13E In ATTR patients, IVS thickness (Y-axis) and cardiac SUVR 平均 (X-axis) related ( Figure 13FIn patients with amyloidosis, cardiac LV wall thickness (Y-axis) and cardiac SUVR... 平均 (X-axis) related ( Figure 13G In patients with amyloidosis, GLS (Y-axis) and cardiac SUVR 平均 (X-axis) related ( Figure 13H In patients with amyloidosis, IVS thickness (Y-axis) is correlated with cardiac SUVR. 平均 (X-axis) related ( Figure 13I ). Invention Details

[0067] All publications mentioned in this application, including patent literature, scientific articles, and databases, are incorporated herein by reference to the same extent that each individual publication is individually incorporated by reference. Where the definitions described herein contradict or otherwise are inconsistent with those described in patents, applications, publications, and other publications incorporated herein by reference, the definitions described herein shall prevail over those incorporated herein by reference.

[0068] The chapter titles used in this article are for organizational purposes only and should not be construed as limiting the topics described. I. Overview

[0069] This document provides methods for diagnosing and treating amyloidosis. In some embodiments, the methods include administering an amyloid reactive agent and detecting the amount of amyloid reactive peptides by detecting the amount of a detectable marker in an individual's tissues or organs. In some embodiments, the methods provided herein are capable of diagnosing amyloid-related disease in individuals at risk of developing it. In some embodiments, the methods provided herein are capable of diagnosing amyloid-related disease in individuals suspected of having it. In some embodiments, the methods provided herein are capable of determining the prognosis of an individual diagnosed with amyloid-related disease. In some embodiments, the methods provided herein are capable of monitoring treatment in individuals with amyloid-related disease. In some embodiments, the methods provided herein are capable of selecting treatment for individuals with amyloid-related disease.

[0070] Early mortality in newly diagnosed amyloid-related diseases indicates a significant delay in diagnosis (Muchtar E, Gertz MA, Kumar SK, et al.Blood.2017; 129(15):2111-2119). For example, nearly 20% of patients die from AL amyloidosis within 6 months of diagnosis, and this statistic has not shown any improvement over 40 years, suggesting that despite significant advances in the treatment of the disease, patients diagnosed at an advanced stage are not being helped (Hasib Sidiqi, M, and Morie A Gertz. Blood cancer journal vol. 11,590. 15 May. 2021). Once a diagnosis of amyloidosis has been established, current anatomical imaging methods, including echocardiography and cardiac magnetic resonance imaging, provide valuable structural and functional information. 99m Tc-PyP and similar bone-finding agents are used to detect cardiac amyloid; however, their relationship with amyloid fibrils remains unclear. Therefore, a non-invasive imaging approach using reagents that directly bind to amyloid deposits is quantifiable and could detect cardiac amyloidosis before significant anatomical changes and organ dysfunction occur, which would have significant clinical benefits.

[0071] In some embodiments, this document provides a method for diagnosing amyloid-associated disease in an individual at risk of developing such disease, comprising administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs, wherein an amount of the amyloid-reactive peptide above a threshold indicates that the individual has amyloidosis.

[0072] This also provides, in some embodiments, a method of treating amyloid-related diseases, comprising administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label in the individual's tissues or organs; and administering treatment for amyloid-related diseases if the amount of the amyloid-reactive peptide is above a threshold. II. Definition

[0073] As used herein, “amino acid” or “amino acid residue” refers to any naturally occurring amino acid, any non-naturally occurring amino acid, any modified (including derived) amino acid, or any amino acid analogue known in the art. Amino acids can be referred to by their common three-letter abbreviations and single-letter abbreviations.

[0074] The terms amyloid protein, amyloid deposits, amyloid fibrils, and amyloid filaments refer to insoluble fibrin aggregates sharing specific structural features. These protein aggregates possess a tertiary structure; for example, they can be formed by the aggregation of any of several different proteins and consist of an ordered arrangement of β-sheets stacked perpendicular to the fibrous axis. See Sunde. et al. J. Mol. Biol. (1997) 273:729-39. Abnormal accumulation of amyloid proteins in organs can lead to amyloidosis. Although their appearance differs, all amyloid proteins share common morphological properties because they are stained with specific dyes such as Congo red and exhibit a characteristic red-green birefringent appearance in polarized light after staining. Amyloid proteins also share common ultrastructural features and common X-ray diffraction and infrared spectra.

[0075] Amyloidosis refers to a pathological condition or disease characterized by the presence of amyloid proteins (e.g., amyloid deposits). "Amyloidosis" or "amyloid degeneration" is a disease associated with the formation, deposition, accumulation, or persistence of amyloid fibrils. These diseases include, but are not limited to, Alzheimer's disease, Down syndrome, hereditary cerebral hemorrhage with Dutch amyloidosis, and cerebral β-amyloid angiopathy. Other amyloidosis, such as systemic AA amyloidosis, AL amyloidosis, ATTR amyloidosis, ALECT2 amyloidosis, and IAPP amyloidosis in type II diabetes, are also amyloidosis.

[0076] The term "amyloid-forming" refers to proteins that produce or are prone to producing amyloid deposits. For example, certain soluble monomeric proteins can undergo extensive conformational changes, leading to their aggregation into ordered, unbranched fibrils 8 to 10 nm wide, which ultimately form amyloid aggregates. More than thirty proteins have been found to form amyloid deposits (or amyloids) in humans. Not all proteins within this category, such as immunoglobulin light chains, are capable of forming amyloids; some immunoglobulin light chains are non-amyloid-forming, meaning they do not tend to form amyloids. However, other proteins in this category can form amyloid deposits and are therefore amyloid-forming. Furthermore, within the category of light chain proteins, some can be considered more "amyloid-forming" than others based on their ease of forming amyloid fibrils. Some light chain proteins are considered non-amyloid-forming or less amyloid-forming because they do not readily form amyloid fibrils in patients or in vitro.

[0077] As used herein, the term "carrier" includes pharmaceutically acceptable carriers, excipients, or stabilizers that are non-toxic to cells, tissues, mammals, or subjects exposed to them at the doses and concentrations used. Typically, pharmaceutically acceptable carriers are aqueous pH-buffered 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 about 10 residues) peptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as 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®.

[0078] As used herein, the term "effective amount" or "appropriate amount" is an amount sufficient to achieve a beneficial or desired clinical or biochemical outcome. An effective amount may be administered once or multiple times. For the purposes of this invention, an effective amount of amyloid reactant or detector is an amount sufficient to bind and allow for the detection of amyloid.

[0079] As used herein, the terms “imaging agent” or “contrast agent” (these terms are used interchangeably) refer to any reagent that can be used in conjunction with methods for imaging internal regions of a subject by applying and / or detecting an energy source and / or for diagnosing the presence or absence of disease in the subject. Exemplary imaging agents include contrast agents used in conjunction with ultrasound, magnetic resonance imaging, radionuclide imaging, or X-ray (including computed tomography) imaging of a patient, as well as the compositions described herein.

[0080] As used herein, the term "mammal" for the purposes of this invention means any animal classified as a mammal, including humans, livestock and farm animals, as well as zoo, sport or pet animals, such as dogs, cats, cattle, horses, sheep, pigs, etc. In some embodiments, the mammal is a human.

[0081] As used herein, the term “peptide” refers to any peptide or peptide analog structure that contains two or more amino acids or is composed of two or more amino acids, including chemical modifications and derivatives of amino acids.

[0082] As used herein, “purified” or “isolated” molecules refer to biomolecules or synthetic molecules that have been removed from their natural environment and separated or isolated, and that do not contain other components associated with their natural environment.

[0083] As used herein, the term "specific binding" refers to a non-random binding reaction between two molecules, such as between an amyloid reactant and amyloid protein. The term "specific binding" may be used interchangeably with "selective targeting" or "selective association."

[0084] As used herein, the terms "selective targeting" or "selective association" refer to, for example, the selective localization or binding between an amyloid reactant and amyloid proteins, relative to non-amyloid proteins. Amyloid reactants can selectively target multiple types of amyloid proteins.

[0085] As used in this article, the term "subject" refers to a vertebrate. A vertebrate can be a mammal, such as a human. A subject can be a human patient.

[0086] As used in this article, the term "amyloid reactant" refers to a reagent that specifically reacts with or binds to amyloid protein.

[0087] As used in this article, the term "combined" includes both covalent and non-covalent combinations.

[0088] The term “antibody” is used in the broadest sense in this article and specifically covers monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, provided they exhibit the desired biological activity.

[0089] =The term “amyloid reactive peptide” refers to a peptide that binds to any of the various amyloid fibrils, such as, but not limited to, amyloidogenic λ6 variable domain protein (Vλ6Wil) or amyloidogenic immunoglobulin light chain (AL), Aβ (1-40) amyloid-like fibrils or amyloidogenic AP precursor protein, or serum amyloid A (AA). In other embodiments, the amyloid protein bound by the amyloid reactive peptide includes the following amyloid-generating forms: immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin variant (ATTRv), wild-type transthyretin (ATTRwt), apolipoprotein A1 (AApoAI), apolipoprotein A1I (AApoAII), colloidin (AGel), lysozyme (ALys), leukocyte chemokine (ALect2), fibrinogen α variant (AFib), cystatin variant (ACys), calcitonin (Aca1), lactoglucosin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (Alns), prion protein (APrP); α-synuclein (AaSyn), tau (ATau), atrial natriuretic factor (AANF), or other amyloid-generating peptides such as IAAP, ALκ, and ALλ1. II. Diagnostic methods

[0090] This article provides a method for diagnosing amyloid-related disease in individuals at risk of developing the disease, comprising administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs, wherein an amount of the amyloid-reactive peptide above a threshold indicates that the individual has amyloidosis.

[0091] This article provides a method for early diagnosis of amyloid-associated disease in an individual, comprising administering an amyloid-reactive peptide conjugated to a detectable marker to the individual, and detecting the amount of the detectable marker in the individual's tissues or organs, wherein an amount of the amyloid-reactive peptide above a threshold indicates that the individual has amyloid-associated disease, wherein the amyloid-associated disease is in its early stages. In some embodiments, the amyloid-associated disease is systemic amyloidosis. In some embodiments, the amyloid-associated disease is cardiac amyloidosis. In some embodiments, the amyloid-reactive peptide conjugated to the detectable marker is a peptide containing... 124 A peptide of the amino acid represented by SEQ ID NO: 100 conjugated with I. In some embodiments,124 I binds to amino acid 4 of SEQ ID NO: 100.

[0092] The methods provided herein can be used for the early diagnosis of amyloid-related diseases. In some embodiments, the methods can be used to diagnose amyloid-related diseases prior to conventional diagnostic methods. In some embodiments, the individual has been previously diagnosed as not having amyloid-related diseases. In some embodiments, the methods have high sensitivity and specificity for the early diagnosis of amyloid-related diseases. A. Amyloid reactive peptides

[0093] In some implementations, methods for diagnosing the type of amyloidosis include administering an amyloid reactive peptide conjugated to a detectable marker.

[0094] In some embodiments, a method for diagnosing the type of amyloidosis includes administering an amyloid reactive agent comprising an amyloid reactive peptide conjugated to a detectable marker. In some embodiments, the amyloid reactive peptide comprises at least 80%, 85%, 90%, or higher of the same amino acid sequence as any one of the amino acid sequences shown in SEQ ID NO: 1-14, for example, having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% the same amino acid sequence as any one of the amino acid sequences shown in SEQ ID NO: 1-14. In some embodiments, the amyloid reactive peptide used with the methods described herein comprises or consists of about 10 to 55 amino acids. The amyloid-reactive peptide of the present invention may, for example, comprise or consist of 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 amyloidosis includes an amyloid-reactive peptide having the amino acid sequence shown in SEQ ID NO: 13. In some embodiments, a method for diagnosing the type of amyloidosis includes p5+14. In some embodiments, the method for diagnosing amyloidosis includes having with 124 I-conjugated amyloid reactive peptide with the amino acid sequence shown in SEQ ID NO: 13. Table 1. Examples of amyloid reactive peptide sequences

[0095] The amino acids forming all or part of the amyloid reactive peptides used in the methods of the present invention can be naturally occurring amino acids, non-naturally occurring amino acids, post-translational modified amino acids, enzymatically synthesized amino acids, derived amino acids, stereoisomers and modifications designed to mimic the structure of amino acids, etc. The amino acids forming the peptides of the present invention can be one or more of the 20 common amino acids found in naturally occurring proteins, or one or more of modified and uncommon amino acids. The amyloid reactive peptides used with the methods described herein can be prepared by any technique known to those skilled in the art, including chemical synthesis or recombinant methods using standard molecular biology techniques.

[0096] The peptides of the present invention may also comprise one or more modified amino acids. Modified amino acids may be derived amino acids or modified and uncommon amino acids. Examples of modified and uncommon amino acids include, but are not limited to, 2-aminohexanoic acid (Aad), 3-aminohexanoic acid (Baad), β-aminopropionic acid (Bala, β-alanine), 2-aminobutyric acid (Abu, piperidinic acid), 4-aminobutyric acid (4Abu), 6-aminohexanoic acid (Acp), 2-aminoheptanoic acid (Ahe), 2-aminoisobutyric acid (Aib), 3-aminoisobutyric acid (Baib), 2-aminopimelic acid (Apm), 2,4-diaminobutyric acid (Dbu), desmokinin (Des), 2,2'-diaminopimelic acid (Dpm), 2,3-diaminopropionic acid (Dpr), N-ethylglycine (EtGly), and N-ethylasparagine (EtGly). Hydroxylysine (Hyl), allo-hydroxylysine (AHyl), 3-hydroxyproline (3Hyp), 4-hydroxyproline (4Hyp), isodesin (Ide), allo-isoleucine (Alle), N-methylglycine (MeGly, sarcosine), N-methylisoleucine (Melle), 6-N-methyllysine (MeLys), N-methylvaline (MeVal), orthovaline (Nva), ortholeucine (Nle), and ornithine (Orn).

[0097] Other examples of modified and less common 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; the teachings of all these are incorporated herein by reference.

[0098] In some embodiments, the peptide of the present invention may comprise or consist of the following amino acid sequence: SRAQRAQARQARQAQRAQRAQARQARQ. (SEQ ID NO: 17).

[0099] The peptides of the present invention can be fusion proteins containing a second peptide as a leader sequence at the amino terminus, such as CGGY (SEQ ID NO: 18) or GGGYS (SEQ ID NO: 19), labeled with a detection reagent. Therefore, in some embodiments, the amyloid-reactive peptide can have up to 55 amino acids and contain the amino acid sequence shown in CGGYSRAQRAQARQARQAQRAQRAQARQARQ (SEQ ID NO: 20).

[0100] In some embodiments, this document provides a peptide comprising an amyloid-reactive peptide fused with 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 document provides a peptide comprising an amyloid-reactive peptide and the N-terminal leader sequence GGGYS, wherein the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-14. In some embodiments, this document provides a peptide comprising the amino acid sequence GGGYS KAQKA QAKQA KQAQK AQKAQ AKQAK QAQKAQKAQA KQAKQ (SEQ ID NO: 100). In some embodiments, the peptide comprises the amino acid sequence shown in SEQ ID NO: 100 and is conjugated to a detectable tag. In some embodiments, the detectable tag binds to one or more of amino acids 1-6 of SEQ ID NO: 100. In some embodiments, the detectable tag binds to amino acid 4 of SEQ ID NO: 100. In some embodiments, the marker can be detected to bind to the N-terminal leader sequence. In some embodiments, the peptide comprises the amino acid sequence shown in SEQ ID NO: 100 and is conjugated to... 124 I can detect markers. In some implementations, 124 I can detect the binding of the marker to one or more of amino acids 1-6 of SEQ ID NO: 100. In some embodiments, 124 The detectable marker binds to amino acid 4 of SEQ ID NO: 100. In some embodiments, the detectable marker binds to the N-terminus.

[0101] Fusion proteins may contain other leader sequences, such as cell-penetrating peptides (CPP) or blood-brain barrier (BBB) ​​translocating peptides.

[0102] The present invention also provides other peptides and fusion proteins rich in positively charged amino acids for imaging amyloid proteins.

[0103] The peptides of this invention can be prepared by any technique known to those skilled in the art, including chemical synthesis, recombinant methods using standard molecular biology techniques, or isolation of peptides from natural sources. Peptides can be synthesized in solution or on a solid support according to conventional techniques. 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, Gross and Meienhofer, eds., Academic Press, New York, pp. 1-284, 1979, which are each incorporated herein by reference in their entirety.

[0104] Alternatively, recombinant DNA technology can be used, wherein 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.

[0105] In some embodiments, the amyloid reactant can be a naturally occurring peptide and can be obtained by isolating or purifying it from its natural source. Protein purification techniques involve, at one level, the homogenization and coarse fractionation of cells, tissues, or organs into peptide and non-peptide fractions. Other protein purification techniques include, for example, precipitation with ammonium sulfate, polyethylene glycol (PEG), antibodies, etc., or by heat denaturation followed by: centrifugation; chromatographic steps, such as ion exchange, gel filtration, reversed-phase chromatography, hydroxyapatite, and affinity chromatography; isoelectric focusing; gel electrophoresis, such as polyacrylamide gel electrophoresis; and combinations of these and other techniques.

[0106] Various chromatography techniques include, but are not limited to, ion exchange chromatography, gel size exclusion chromatography, affinity chromatography, immunoaffinity chromatography, and reversed-phase chromatography. A particularly efficient method for purifying peptides is rapid high-performance liquid chromatography (FPLC) or even high-performance liquid chromatography (HPLC).

[0107] The order of various purification steps can be changed, or certain steps can be omitted, while still producing a suitable method for preparing substantially purified peptides. The peptides of the present invention can be part of a polypeptide or protein and can be produced by the biochemical or enzymatic fragmentation of the polypeptide or protein. Therefore, the peptides of the present invention can be (a) naturally occurring, (b) produced by chemical synthesis, (c) produced by recombinant DNA technology, (d) produced by the biochemical or enzymatic fragmentation of larger molecules, (e) produced by a method combining methods a to d above, or (f) produced by any other means for producing peptides.

[0108] During chemical synthesis, peptides can be modified at their N- or C-termini to provide improved stability and formulation, resistance to protease degradation, and more. Examples of amino acid modifications include polyethylene glycolation, acetylation, alkylation, formylation, and amidation. Furthermore, a variety of non-naturally occurring amino acids can be introduced along the chain to improve peptide stability.

[0109] Cysteine ​​is also used to facilitate the labeling of the peptides of the present invention via conjugation with biotin, fluorophores, or other ligands. Furthermore, cysteine ​​residues on the leader peptide allow for the formation of covalently bound dimer molecules, which can increase the relative affinity of the peptide for its target. B. Detectable Markings

[0110] In some embodiments, methods for diagnosing the type of amyloidosis include applying an amyloid-reactive peptide conjugated to a detectable label. The conjugation, as used herein, can be covalent and / or non-covalent. In some embodiments, the detectable label is covalently or non-covalently bound to the peptide. Without limitation, this can include 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; fluorescent groups, etc. Any means known in the art for detectably labeling proteins can be used and / or adapted to the methods described herein. For example, the amyloid-reactive peptide can be radiolabeled with a radioisotope, or labeled with a fluorescent tag or chemiluminescent tag. Example radioisotopes include, for example, 11 C 18 F, 111 In、 99m Tc and 123 I, 124 I and 125 I. These and other radioactive isotopes can be incorporated into amyloid reactants or detection dyes. Example fluorescent or chemiluminescent tags include fluorescein, Texas red, rhodamine, Alexa dye, and luciferase, which can be incorporated into amyloid reactants using methods conventional in the art.

[0111] In some embodiments, methods for diagnosing the type of amyloidosis include administering an amyloid reaction agent containing a radiolabeled substance. In some embodiments, the radiolabeled substance is... 11 C 18 F, 111 In、 99m Tc, 89 Zr and 123 I, 124 I or 125I. In some embodiments, the radiolabeled amyloid reactive agent is a radiolabeled amyloid reactive peptide. In some embodiments, the radiolabeled amyloid reactive peptide is... 124 I-labeled amyloid reactive peptides. In other embodiments, methods for diagnosing the type of amyloidosis include administering... 124 I-p5+14. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence shown in SEQ ID NO: 13. In some embodiments, the amyloid-reactive peptide is related to... 124 I. Conjugation. In some embodiments, methods for diagnosing the type of amyloidosis include administering an agent containing... 124 I-conjugated amyloid reactive peptide of SEQ ID NO: 13.

[0112] In another embodiment, a method for diagnosing the type of amyloidosis includes administering a fluorescently labeled amyloid reagent. In some embodiments, the fluorescently labeled amyloid reagent is thioflavin T (ThT).

[0113] In some embodiments, the amyloid reactive agent comprises an amyloid reactive peptide conjugated to a radiolabel. In some embodiments, the amyloid reactive agent comprises a peptide conjugated to an extender. In some embodiments, the amyloid reactive peptide is conjugated to PEG. In some embodiments, the amyloid reactive peptide is conjugated to an antibody.

[0114] In some embodiments, the amyloid reactant specifically binds to amyloid deposits. In some embodiments, the amyloid reactant is capable of detecting the presence, absence, or amount of amyloid in a subject. In some embodiments, the amyloid reactant or dye cross-reacts with amyloid deposits formed from many different proteins. In some embodiments, the amyloid reactant binds to amyloid deposits formed from multiple proteins and / or peptides. In some embodiments, the amyloid reactant binds to amyloid deposits formed from amyloid light chains (AL). In some embodiments, the amyloid reactant binds to amyloid formed from transthyretin (TTR) fibrils. In some embodiments, the amyloid reactant binds to amyloid formed from serum amyloid A (sAA). In some embodiments, the amyloid reactant binds to the following amyloid-generating forms: 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), colloidin (AGel), lysozyme (ALys), leukocyte chemokine (ALECT2), fibrinogen a variant (AFib), cystatin variant (ACys), calcitonin (ACa1), lactoglucosin (AMed), islet amyloid peptide (AIAPP), prolactin (APro), insulin (AIns), prions (APrP); α-synuclein (AαSyn), tua (ATau), atrial natriuretic factor (AANF), or other amyloid-generating peptides such as IAAP, ALκ4, and ALλ1.

[0115] In some embodiments, the amyloid reactant is bound to heparan sulfate glycosaminoglycan (GAG). In some embodiments, GAG is associated with amyloid deposits. The binding of GAG to amyloid fibrils occurs primarily through electrostatic interactions involving negatively charged polyelectrolyte charges and positively charged side-chain residues of the aggregated protein. Similar to catalysts used in the reaction, GAG promotes aggregation, nucleation, and amyloid fibril formation, functioning as a structural template for the self-assembly of highly cytotoxic oligomeric precursors rich in β-sheets into amyloid fibrils. Furthermore, the specific interaction at the shared binding site between amyloid peptides and GAG molecules can contribute to the GAG-mediated amyloid activity.

[0116] In some embodiments, the method includes administering an amyloid-reactive agent to an individual. In some embodiments, the amyloid-reactive agent is administered as a pharmaceutical composition. In some embodiments, the composition comprises an aqueous buffer. The composition may also include a solubilizer and a local anesthetic such as lidocaine to relieve pain at the injection site. The components are provided individually or mixed together in unit dosage forms, for example, as a dried lyophilized powder or anhydrous concentrate in a sealed container such as an ampoule indicating the amount of active agent. When the composition is to be administered by infusion, it can be dispensed using an infusion bottle containing sterile pharmaceutical-grade water or saline. When the composition is to be administered by injection, ampoules of sterile water for injection or saline can be provided so that the components can be mixed prior to administration.

[0117] 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 and detecting amyloid protein and a pharmaceutically acceptable carrier.

[0118] Pharmaceutically viable carriers include solid or liquid carriers or components that can be added to enhance or stabilize a composition or to facilitate its preparation, including but not limited to syrups, water, isotonic saline solutions, 5% glucose solutions, or buffered sodium or ammonium acetate solutions, oils, glycerol, alcohols, etc. 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 include sustained-release materials, such as glyceryl monostearate or glyceryl distearate alone or with waxes. Other suitable drug carriers include, but are not limited to, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, propylene, ethylene glycol, water, ethanol, flavoring agents, preservatives, coloring agents, diluents, granulating agents, lubricants, binders, etc.

[0119] When administering a drug composition intravenously, water is a preferred carrier. Saline, glucose, and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. The composition may also contain small amounts of wetting agents, emulsifiers, or pH buffers, if desired. Such compositions can be in the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations, etc. The composition can be formulated into suppositories using conventional binders and carriers such as triglycerides. Oral formulations may include standard carriers such as pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Examples of other suitable drug carriers are described in EW Martin's "Remington's Pharmaceutical Sciences".

[0120] Methods for imaging amyloid protein include, but are not limited to, magnetic resonance imaging (MRI), computed axial computed tomography (CAT) scans, positron emission tomography (PET), ultrasound imaging, X-ray, radionuclide imaging, single-photon emission computed tomography (SPECT), and multiphoton microscopy.

[0121] To increase the sensitivity of a scan, various contrast agents can be used. Contrast agents used for scanning can include all molecules that attenuate X-rays. For positron emission tomography (PET) and radionuclide imaging, radioisotopes can be used. All positron emission isotopes can be used for PET radionuclide imaging, and all gamma-photon emission isotopes can be used for radionuclide imaging.

[0122] Contrast agents used for ultrasound imaging include positive and negative agents. Positive agents reflect ultrasound energy, thus producing positive (bright) images. Correspondingly, negative agents enhance transmissibility or acoustic transmission, thus producing negative (dark) images. A variety of substances—gases, liquids, solids, and combinations thereof—have been investigated as potential contrast enhancers. Examples of solid particulate contrast agents are disclosed in U.S. Patent No. 5,558,854, including, but not limited to, IDE particles and SHU454. European Patent Application 0231091 discloses oil-in-water emulsions containing highly fluorinated organic compounds for providing enhanced contrast in ultrasound images. Emulsions containing perfluorooctane bromide (PFOB) have also been tested as ultrasound imaging agents. U.S. Patent No. 4,900,540 describes the use of gases or gaseous precursors containing phospholipid-based liposomes as contrast enhancers.

[0123] Several classes of compounds have the potential to serve as MRI contrast agents. These classes include superparamagnetic iron oxide particles, nitro oxides, and paramagnetic metal chelates (Mann et al., 1995). Strongly paramagnetic metals are preferred. Typically, paramagnetic lanthanides and transition metal ions are toxic in vivo. Therefore, it is necessary to conjugate these compounds with organic ligands into chelates. The peptides and fusion peptides of the present invention can be used to enhance the targeting of such chelated metals to amyloid proteins, which allows for a reduction in the total dose originally required to image the composition.

[0124] Developers can be attached to peptides and fusion peptides using known methods. Some attachment methods involve the use of metal chelating complexes, such as organic chelating agents like DTPA. Acceptable chelates are known in the art. They 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 other chelates.

[0125] Several classes of compounds have the potential to serve as MRI contrast agents. These classes include superparamagnetic iron oxide particles, nitro oxides, and paramagnetic metal chelates (Mann et al., 1995). Strongly paramagnetic metals are preferred. Typically, paramagnetic lanthanides and transition metal ions are toxic in vivo. Therefore, it is necessary to conjugate these compounds with organic ligands into chelates. The peptides and fusion peptides of the present invention can be used to enhance the targeting of such chelated metals to amyloid proteins, which allows for a reduction in the total dose originally required to image the composition.

[0126] 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 available in other applications (e.g., X-ray imaging) include, but are not limited to, lanthanum(III), gold(III), lead(II), and especially bismuth(III).

[0127] Radioactive isotopes that can be used to label the peptides and fusion peptides of the present invention suitable for localization studies include gamma emitters, positron emitters, X-ray emitters, and fluorescent emitters. Suitable radioactive isotopes 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 ,gallium67 ,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 , 32 Phosphorus, 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 Yttrium 90 Halogens can be used more or less interchangeably as labelling agents. Gamma-emitting iodine can also be used. 123 Hété 99m This is because such radioactive metals are detectable with gamma cameras and have a favorable half-life for in vivo imaging. Alternatively, positron emitters suitable for PET imaging and having a suitable half-life for peptide imaging, such as 18-fluoro or... 124 Iodine. The peptides and fusion peptides of the present invention can be labeled with indium 111 or technetium 99m via a conjugated metal chelating agent such as DTPA (diethylenetriaminepentaacetic acid) or non-covalently and directly to flanking peptides containing Cys residues or triglycine motifs.

[0128] Radiolabeled peptides or fusion peptides can be produced according to methods well known in the art. For example, they can be iodinated by contact with sodium or potassium iodide and a chemical oxidant such as sodium hypochlorite or an enzymatic oxidant such as lactoperoxidase. The peptides or fusion peptides according to the invention can be produced using technetium via a ligand exchange process. 99mLabeling can be performed, for example, by reducing pertechnate with a stannous solution, chelating the reduced technetium onto a Sephadex column, and then applying the peptide to the column, or by direct labeling techniques, such as incubating the pertechnate, a reducing agent such as SnCl2, a buffer solution such as sodium potassium phthalate solution, and the peptide. As previously mentioned, the intermediate functional groups commonly used to bind radioactive isotopes, which are present in the form of metal ions, to peptides are diethylenetriaminepentaacetic acid (DTPA) and ethylenediaminetetraacetic acid (EDTA).

[0129] Other useful labels include fluorescent labels, chromogenic labels, and biotin labels. Fluorescent labels include, but are not limited to, rhodamine, fluorescein isothiocyanate, sodium fluorescein, renal contrast agents, and Texas erythromylate chloride. In some embodiments, the peptides and fusion peptides of the present invention may be linked to a secondary binding ligand or an enzyme (enzyme tag) that will generate a colored product upon contact with a chromogenic substrate. Examples of suitable enzymes include urease, alkaline phosphatase, (horseradish) catalase, and glucose oxidase. Secondary binding ligands include biotin and avidin or streptavidin compounds. The use of such labels is well known to those skilled in the art, as described, for example, in U.S. Patent Nos. 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149, and 4,366,241, each of which is incorporated herein by reference.

[0130] This invention provides a method for detecting amyloid protein in a subject. The method includes administering a pharmaceutical composition comprising an effective amount of one or more peptides or fusion peptides of the present invention to the subject, and detecting the peptides or fusion peptides that bind to amyloid protein. The peptides can be labeled with imaging agents such as radioisotopes. The peptides have a specific binding affinity for deposits, and the binding is detectable. The binding of the peptides or fusion peptides to amyloid protein can be detected by MRI, CAT scan, PET imaging, ultrasound imaging, SPECT imaging, X-ray imaging, fluorescence imaging, or radionuclide imaging.

[0131] In some cases, methods for diagnosing the type of amyloidosis include administering a detectable amount of amyloid reactant or dye to an individual. The detectable amount to be administered can be based on the type of test to be performed. For example, in some embodiments, the detectable amount of amyloid reactant or dye can be an amount sufficient to be detected by imaging when administered to a subject. The detectable amount of amyloid reactant or test dye to be administered to an individual can vary depending on factors such as the individual's age, sex, and weight, the individual's specific response, dosing, formulation, and instrument-related factors. Optimization of these factors is entirely within the scope of the art. The detectable amount of amyloid reactant can also vary depending on the mode of administration of the amyloid reactant or test dye.

[0132] In some cases, amyloid reactants are administered parenterally, adjacent to cancer, transmucosal, transcutaneous, intramuscular, intravenous, intradermal, subcutaneous, intraperitoneal, intravenous, or intracranial. In other cases, amyloid reactants are administered intravenously. In still other cases, amyloid reactants are administered intraperitoneally.

[0133] Those skilled in the art will further understand that an effective amount of amyloid reactant can be administered in a single dose or by administering multiple doses. In some cases, administration of the amyloid reactant may further include administration of a flushing solution. For example, a flushing solution such as saline may be administered immediately after administration of the amyloid reactant or the detection dye, or some time after administration of the amyloid reactant or the detection dye. In other cases, the amyloid reactant may be metabolized and excreted over a specific period of time following administration. C. Detection of amyloid protein in organs and tissues

[0134] In some embodiments, methods for diagnosing amyloidosis include detecting amyloid protein using an amyloid protein reactant or detection dye. Examples of amyloid proteins that can be detected as part of this method include, but are not limited to, the following amyloid-producing forms: immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin variant (ATTR), amyloid β (Aβ), apolipoprotein AI (AApoAI), apolipoprotein AII (AApoAII), colloidin (AGel), lysozyme (ALys), leukocyte chemokine (ALect2), fibrinogen α variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactoglucosin (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-producing peptides. In some embodiments of this disclosure, methods for diagnosing the type of amyloidosis include detecting ATTR, AL, and / or ALECT2 amyloid proteins. In other embodiments, methods for diagnosing the type of amyloidosis include differentiating between ATTR, AL, and / or ALECT2 amyloid proteins.

[0135] In some embodiments, the method can be used to diagnose amyloid-related diseases. In some embodiments, amyloid-related diseases are AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AIAPP, APro, Alns, 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.

[0136] In some embodiments, methods for diagnosing amyloid-related diseases include administering an amyloid reactive peptide conjugated to a detectable marker to an individual and measuring the amount of the detectable marker in the individual's tissues or organs, wherein an amount of signal from the amyloid reactive peptide above a threshold indicates that the individual has amyloidosis.

[0137] In some embodiments, the method includes administering to an individual a dose of about 0.1 mCi to about 25 mCi of a detectable label-conjugated amyloid-reactive peptide. In some embodiments, the method includes administering to an individual a dose of about 0.3 mCi, 1 mCi, or about 2 mCi of a detectable label-conjugated amyloid-reactive peptide. In some embodiments, the method includes administering to an individual a dose of 1 mCi (±10%) of a detectable label-conjugated amyloid-reactive peptide. In some embodiments, the method includes administering to an individual a dose of about 0.9 mCi to about 1.1 mCi of a detectable label-conjugated amyloid-reactive peptide. In some embodiments, the method includes administering to an individual a dose of 1 mCi (±10%) and not exceeding 2 mCi of a detectable label-conjugated amyloid-reactive peptide. In some embodiments, the detectable label-conjugated amyloid-reactive peptide is... 124 I-labeled amyloid reactive peptide.

[0138] In some embodiments, the method includes administering to an individual a dose of about 0.1 mCi to about 25 mCi. 124 I-labeled amyloid reactive peptide. In some embodiments, the method includes administering to an individual a dose of about 0.3 mCi, 1 mCi, or about 2 mCi. 124 I-labeled amyloid reactive peptide. In some embodiments, the method includes administering to an individual a dose of 1 mCi (±10%). 124 I-labeled amyloid reactive peptides. In some embodiments, the method includes administering to an individual a dose of about 0.9 mCi to about 1.1 mCi. 124 I-labeled amyloid reactive peptides. In some embodiments, the method includes administering to an individual a dose of 1 mCi (±10%) and not exceeding 2 mCi. 124 I-labeled amyloid reactive peptide.

[0139] In some embodiments, the method includes administering to 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 includes administering to 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 includes administering to an individual about 0.3 mg to about 1 mg or about 0.3 mg to about 1.5 mg of amyloid-reactive peptide.

[0140] In some embodiments, the step of detecting amyloid reactants in one or more organs in this method includes determining an organ uptake value for each organ. Organ uptake can be determined by methods known to those skilled in the art. For example, an organ uptake value can indicate the relative or absolute level of amyloid reactants detected in each organ of an individual. In some embodiments, the organ uptake value ratio is a relative uptake value. In some embodiments, the organ uptake value is a standard uptake value (SUV) for each organ. In some embodiments, as those skilled in the art will understand, the standard uptake value can be determined by measuring the amount of amyloid reactants detected in a reactive organ (e.g., the heart) relative to the amount of amyloid reactants detected in a non-reactive tissue or sample (e.g., blood). The amount of amyloid reactants in an organ can be determined, for example, by quantifying a detectable signal from the amyloid reactants in the organ, such as by calculating pixel values ​​in an image. In some embodiments, the standard uptake value is determined as the ratio of the amount of amyloid reactants detected in an organ to the amount of amyloid reactants detected in blood. In some embodiments, the organ uptake value indicates the organ distribution pattern of the amyloid reactants or detection dye. In some embodiments, the method includes determining an organ-specific SUV for an individual. In some embodiments, the organ-specific SUV for an individual is selected from a group consisting of the average SUV, the maximum SUV, and the peak SUV.

[0141] In some embodiments, organ uptake is based on the organ-specific standardized uptake ratio (SUVR). In some embodiments, a blood pool is used as a reference tissue to calculate the SUVR. In some embodiments, the SUVR for each organ is calculated by dividing the amount of amyloid detection agent 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 a group consisting of the SUVR mean, SUVR maximum, 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 control for differences in radioactive tracer clearance rates.

[0142] In some embodiments, the threshold is an average SUVR threshold. In some embodiments, the average SUVR threshold is based on the average SUVR threshold of an organ or tissue. The average SUVR threshold can be selected based on the organ or tissue in which amyloid reactive peptides conjugated with detectable markers are detected. In some embodiments, the organ or tissue is the liver, spleen, kidney, pancreas, heart, or adrenal gland. In some embodiments, detecting amyloid reactive peptides conjugated with detectable markers above the average SUVR threshold allows for early diagnosis of amyloid-related conditions.

[0143] In some embodiments, the threshold is the average SUVR threshold. In some embodiments, if the organ or tissue is the liver, the average 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 average SUVR threshold is 1.31. In some embodiments, if an individual's liver has an average SUVR higher than the threshold of 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, or 1.6, the individual may have systemic amyloidosis or a type of systemic amyloidosis. In some embodiments, if an individual's liver has an SUVR higher than 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 type of systemic amyloidosis.

[0144] In some embodiments, the threshold is the average SUVR threshold. In some embodiments, if the organ or tissue is the spleen, the average 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 average SUVR threshold is 1.21. In some embodiments, if an individual's average SUVR in their spleen is above the threshold of 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5, the individual may have systemic amyloidosis or a type of systemic amyloidosis. In some embodiments, if an individual's average SUVR in their spleen is above 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 type of systemic amyloidosis.

[0145] In some embodiments, the threshold is the average SUVR threshold. In some embodiments, if the organ or tissue is the kidney, the average 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 average SUVR threshold is 1.88 (or between 1.69 and 2.07). In some embodiments, if the average SUVR in an individual's kidney is above the threshold of 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, or 2.3, the individual may have systemic amyloidosis or a type of systemic amyloidosis. In some embodiments, if the average SUVR in an individual's kidney is above 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 diagnosed with systemic amyloidosis or a type of systemic amyloidosis.

[0146] In some embodiments, the threshold is the average SUVR threshold. In some embodiments, if the organ or tissue is the pancreas, the average 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 average SUVR threshold is 1.40. In some embodiments, if the average SUVR in an individual's pancreas is higher than the threshold of 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, or 1.7, the individual may have systemic amyloidosis or a type of systemic amyloidosis. In some embodiments, if the average SUVR in an individual's pancreas is higher than the threshold of 1.1, 1.2, 1.3, 1.6, or 1.7, the individual is diagnosed with systemic amyloidosis or a type of systemic amyloidosis.

[0147] In some embodiments, the threshold is the average SUVR threshold. In some embodiments, if the organ or tissue is the heart, the average 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 average SUVR threshold is 1.19. In some embodiments, if the average SUVR in an individual's heart is higher than 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 may have systemic amyloidosis or a type of systemic amyloidosis. In some implementations, an individual is diagnosed with systemic amyloidosis or a type of systemic amyloidosis if the average SUVR in the individual's heart is above 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.

[0148] In some embodiments, the threshold is the average SUVR threshold. In some embodiments, if the organ or tissue is the adrenal gland, the average 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 average SUVR threshold is 1.135. In some embodiments, if an individual's average SUVR in the adrenal gland is higher than the threshold of 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, or 1.4, the individual may have systemic amyloidosis or a type of systemic amyloidosis. In some embodiments, if an individual's average SUVR in the adrenal gland is higher than 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 type of systemic amyloidosis.

[0149] In some embodiments, the threshold is the maximum SUVR threshold. In some embodiments, the maximum SUVR threshold is based on the maximum SUVR threshold of an organ or tissue. The maximum SUVR threshold can be selected based on the organ or tissue in which amyloid reactive peptides conjugated with detectable markers are detected. In some embodiments, the organ or tissue is the liver, spleen, kidney, pancreas, heart, or adrenal gland. In some embodiments, detecting amyloid reactive peptides conjugated with detectable markers above the maximum SUVR threshold allows for early diagnosis of amyloid-related conditions.

[0150] In some implementations, the threshold is the maximum SUVR threshold. In some implementations, if the organ or tissue is the liver, the maximum SUVR 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, 12.9. 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 implementations, if the organ or tissue is the liver, the maximum SUVR threshold is 13.26.

[0151] In some implementations, the threshold is the maximum SUVR threshold. In some implementations, if the organ or tissue is the spleen, the maximum SUVR 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 implementations, if the organ or tissue is the spleen, the maximum SUVR threshold is 4.86.

[0152] In some implementations, the threshold is the maximum SUVR threshold. In some implementations, if the organ or tissue is the kidney, the maximum SUVR 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 implementations, if the organ or tissue is the kidney, the maximum SUVR threshold is 9.68.

[0153] In some implementations, the threshold is the maximum SUVR threshold. In some implementations, if the organ or tissue is the pancreas, the maximum SUVR 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 implementations, if the organ or tissue is the pancreas, the maximum SUVR threshold is 9.48.

[0154] In some implementations, the threshold is the maximum SUVR threshold. In some implementations, if the organ or tissue is the heart, the maximum SUVR 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 implementations, if the organ or tissue is the heart, the maximum SUVR threshold is 6.2.

[0155] In some implementations, the threshold is the maximum SUVR threshold. In some implementations, if the organ or tissue is an adrenal gland, the maximum SUVR threshold is 1, 1.1, 1.2, 1.3, 1.4, 1.5, or 1.6. In some implementations, if the organ or tissue is an adrenal gland, the maximum SUVR threshold is 1.31.

[0156] In some embodiments, the step of measuring the organ uptake value or organ uptake ratio of the amyloid reactant in one or more organs includes analyzing imaging data. The imaging data can be generated by any procedure known in the art that allows imaging of the amyloid reactant or dye. For example, the amyloid reactant can be detected by positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), or single-photon emission computed tomography (SPECT). In some embodiments, the amyloid reactant can be detected by a combination of imaging methods, such as PET / CT (PET with parallel computed tomography) or PET / MRI (PET with parallel magnetic resonance imaging). The imaging process can produce one or more images of an individual's observed region. In some embodiments, imaging produces more than one image, which can be combined, superimposed, added, subtracted, color-coded, or otherwise fused and mathematically manipulated by any method known in the art. The resulting images can be digital or analog images, which can be displayed as "hard" images on, for example, printing paper, photographic paper, or film, or as images on a screen (e.g., a video or LCD screen).

[0157] In some implementations, the region of interest (ROI) method is used to analyze PET images. In some implementations, the images are planar images. In some implementations, the images are coronal, axial, or sagittal images.

[0158] In some embodiments, the threshold for diagnosing a specific type of amyloidosis is selected based on desired sensitivity. In some embodiments, the cutoff is selected to provide 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 with a detectable marker is between about 80% and about 100%. In some embodiments, the sensitivity for detecting cardiac amyloid using an amyloid-reactive peptide conjugated with a detectable marker 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 with a detectable marker is about 96%.

[0159] In some embodiments, the sensitivity for detecting cardiac amyloid protein in patients with AL amyloidosis is between about 80% and about 100%. In some embodiments, the sensitivity for detecting cardiac amyloid protein in patients with AL amyloidosis 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 protein in patients with AL amyloidosis is about 93%.

[0160] In some embodiments, the sensitivity for detecting cardiac amyloid protein in patients with ATTR amyloidosis is between about 80% and about 100%. In some embodiments, the sensitivity for detecting cardiac amyloid protein in patients with ATTR amyloidosis 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 protein in patients with ATTR amyloidosis is about 100%.

[0161] In some implementations, the threshold is selected based on the desired specificity (i.e., the ability to exclude individuals without amyloid-related disease). In some implementations, the cutoff is selected to provide at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% specificity.

[0162] In some implementations, a threshold is selected to obtain the optimal relationship between sensitivity and specificity based on organ-specific SUV or SUVR values. In some implementations, the threshold that provides the highest Youden index is selected. In some implementations, the Youden index is determined by the formula: Sensitivity + Specificity - 1. In some implementations, if some SUVs or SUVRs provide the same Youden index, the SUV or SUVR that results in the highest specificity is selected.

[0163] In some implementations, for cardiac SUVR in AL amyloidosis 平均 The threshold is between approximately 0.6 and approximately 4. In some implementations, for cardiac SUVR in AL amyloidosis... 平均 The threshold 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.

[0164] In some implementations, for cardiac SUVR in ATTR amyloidosis 平均The threshold is between approximately 0.6 and approximately 4. In some implementations, for cardiac SUVR in ATTR amyloidosis... 平均 The threshold 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.

[0165] In some embodiments, the sensitivity and specificity of the amyloid-reactive peptide conjugated with a detectable marker are defined relative to clinically approved methods for diagnosing amyloid-related diseases. In some embodiments, clinically approved methods are selected from the group consisting of: measures of an individual's health-related quality of life, 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 the amyloid-reactive peptide conjugated with a detectable marker are defined relative to a diagnosis derived from an individual's medical records.

[0166] In some embodiments, the sensitivity of the amyloid-reactive peptide is the specificity of the amyloid-reactive peptide conjugated with a detectable label. In some embodiments, the sensitivity of the amyloid-reactive peptide conjugated with a detectable label is measured as true positives divided by (true positives plus false negatives). In some embodiments, a true positive is the number of individuals correctly diagnosed with amyloid-related disease using the amyloid-reactive peptide conjugated with a detectable label when the individual actually has the disease. In some embodiments, a false negative is the number of individuals incorrectly diagnosed as not having amyloid-related disease using the amyloid-reactive peptide conjugated with a detectable label when the individual actually has the disease. In some embodiments, sensitivity can be defined as a confidence interval for the sensitivity value. In some embodiments, sensitivity can be defined based on a 95% confidence interval for the sensitivity value. In some embodiments, the Wilson-Brown method can be used to calculate the 95% confidence interval.

[0167] In some embodiments, the sensitivity of the amyloid-reactive peptide conjugated with a detectable marker is the sensitivity for detecting amyloid-related diseases after administration of the amyloid-reactive peptide and detection of the radiolabel by amyloid imaging. In some embodiments, the sensitivity of the amyloid-reactive peptide conjugated with a detectable marker is related to the dose of the amyloid-reactive peptide conjugated with the detectable marker administered to the individual.

[0168] 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 as true negatives divided by (true negatives plus false positives). In some embodiments, a true negative is the number of individuals correctly diagnosed as not having amyloid-related disease based on the amyloid-reactive peptide conjugated to a detectable label, when the individual does not actually have amyloid-related disease. In some embodiments, a false positive is the number of individuals diagnosed as having amyloid-related disease based on the amyloid-reactive peptide conjugated to a detectable label, when the individual does 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 based on a 95% confidence interval of the specificity value. In some embodiments, the Wilson-Brown method may be used to calculate the 95% confidence interval.

[0169] In some embodiments, the specificity of the amyloid-reactive peptide conjugated with a detectable marker is the specificity for detecting amyloid-related diseases after administration of the amyloid-reactive peptide and detection of the radiolabel by amyloid imaging. In some embodiments, the specificity of the amyloid-reactive peptide conjugated with a detectable marker is related to the dose of the amyloid-reactive peptide conjugated with the detectable marker administered to the individual. In some embodiments, the detection step is performed in more than one organ of the individual. In some embodiments, the detection step further includes measuring the organ distribution pattern of the amyloid reactant or detection dye, wherein the organ distribution pattern of the amyloid reactant indicates the type of amyloidosis. The type of amyloidosis may be sporadic amyloidosis or have a genetic component, such as hereditary amyloidosis. Some non-limiting examples of amyloidosis include AA amyloidosis, AL amyloidosis, AH amyloidosis, Aβ amyloidosis, ATTR amyloidosis, ALect2 amyloidosis, and IAPP amyloidosis of type II diabetes, Alzheimer's disease, Down syndrome, hereditary cerebral hemorrhage with Dutch amyloidosis, cerebral β-amyloid angiopathy, spongiform encephalohalopathy, thyroid adenoma, Parkinson's disease, Lewis body dementia, tau disease, Huntington's disease, senile systemic amyloidosis, familial hemodialysis, senile systemic aging, aging-related pituitary disorders, iatrogenic syndromes, spongiform encephalopathies, reactive chronic inflammation, thyroid adenoma, myeloma, or other forms of cancer. In some embodiments, the type of amyloidosis is systemic amyloidosis. In some embodiments, the type of amyloidosis is AL amyloidosis, ATTR amyloidosis, or ALECT2 amyloidosis. In some embodiments, amyloidosis is cardiac amyloidosis. In some embodiments, amyloidosis is cardiac AL amyloidosis. In some embodiments, amyloidosis is cardiac ATTR amyloidosis.

[0170] In some embodiments, methods for diagnosing amyloid-related diseases include measuring the organ distribution pattern of amyloid reactants in one or more organs. Not bound by theory, it is considered that the anatomical distribution of amyloid may have a specific pattern in each form of disease. For example, amyloid deposition in ATTR amyloidosis is prevalent in the heart and peripheral nerves, while AL amyloidosis (another common form of amyloidosis) exhibits a variable pattern of amyloid deposition, with amyloid observed in, for example, the heart, spleen, liver, kidneys, peripheral nerves, gastrointestinal tract, muscles, lungs, and lymph nodes. In some embodiments, methods for diagnosing the type of amyloidosis include measuring the organ distribution pattern of amyloid reactants in one or more of the heart, spleen, liver, kidneys, peripheral nerves, gastrointestinal tract, muscles, lungs, brain, and lymph nodes. In some embodiments, one or more organs are thoracic or abdominal organs. In some embodiments, one or more organs are the heart, spleen, liver, or kidneys.

[0171] In other cases, methods for diagnosing amyloid-related diseases include administering an amyloid reaction agent and calculating organ-to-organ ratios for two or more organs. In some cases, calculating organ-to-organ ratios for two or more organs includes calculating the ratio between organ uptake values ​​for a first organ and organ uptake values ​​for a second organ. In some cases, organ-to-organ ratios are selected from the following groups: liver to heart, spleen to heart, spleen to liver, spleen to kidney, kidney to heart, and kidney to liver. In some cases, the organ-to-organ ratio is the heart to spleen ratio. In some cases, the organ-to-organ ratio is between 0 and 1, 1, or greater than 1. In some cases, organ-to-organ ratios indicate the type of amyloidosis in an individual.

[0172] In some embodiments, organ uptake values ​​or organ-to-organ ratios indicate the type of amyloidosis. In some embodiments, organ uptake values ​​or organ-to-organ ratios only indicate the type of amyloidosis if they are above a cutoff or threshold. For example, in some embodiments, if the organ-to-organ ratio for a type of amyloidosis is 1.4, then a diagnosis of that type of amyloidosis will be made if the organ-to-organ ratio calculated for an individual is 1.4 or higher. As another non-limiting example, if the organ uptake value cutoff for a type of amyloidosis is 1.4, then a diagnosis of that type of amyloidosis is inappropriate if the organ-to-organ ratio calculated for an individual is below 1.4. The specific cutoff or threshold used to diagnose the type of amyloidosis can vary depending on the type of amyloidosis, disease progression, patient demographics, the amyloid reagent administered, and the detection method used. In some embodiments, organ uptake values ​​or organ-to-organ cutoffs or thresholds are calculated from data on organ distribution from amyloid reagents or detection dyes. In some implementations, organ uptake values ​​or organ-to-organ cutoffs or thresholds are calculated from data from a population with a specific type of amyloidosis.

[0173] Preferably, a receiver operating characteristic (ROC) curve is used to determine organ uptake values ​​or organ-to-organ uptake ratio cutoffs or thresholds for diagnosing the type of amyloidosis. As understood in the art, a ROC curve is a graph of the performance of a specific characteristic used to distinguish between two groups, patients with amyloidosis and controls (e.g., those without amyloidosis). Data are sorted in ascending order across the entire population (i.e., patients and controls) based on the values ​​of individual characteristics (e.g., organ uptake values). Then, for each value of that characteristic, the true positive rate and false positive rate of the data are determined. The true positive rate (sensitivity) is determined by counting the number of cases above the value of the characteristic under consideration and then dividing by the total number of patients. The false positive rate (specificity) is determined by counting the number of controls above the value of the characteristic under consideration and then dividing by the total number of controls.

[0174] ROC curves can be generated for a single feature and other individual outputs. For example, combinations of two or more features mathematically added together (addition, plotting, multiplication, etc.) to provide a single total value, which can be plotted in an ROC curve. Furthermore, any combination of multiple features that causes the combination to result in a single output value can be plotted in an ROC curve. These combinations of features can include tests. An ROC curve is a graph of the true positive rate (sensitivity) of a test versus the false positive rate (1-specificity) of a test. The area under the ROC curve can be a figure of merit for a given sample population, ranging from 1 to 0 for a complete test that gives a completely randomized response when classifying test subjects. 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 against another. Using ROC curves, a cutoff value can be selected to diagnose an individual's amyloidosis and / or amyloid type with high confidence.

[0175] In some embodiments, the method includes obtaining organ distribution data for amyloid reactants or detection dyes. In some embodiments, the organ distribution data is an image. Images generated using the imaging process implemented in this invention can be analyzed by any method known in the art. For example, in some embodiments, imaging data derived from PET or SPECT scans can be input into a processor that identifies individual pixels or groups of pixels with brightness greater than a predetermined threshold or average background, and the identified pixels can be characterized as indicating the presence of amyloid reactants or dyes. In another embodiment, image data can be derived from scanned images input into a processor. In such embodiments, a similar process for identifying bright spots in an image can be used to locate amyloid reactants or dyes in the image. In some embodiments, image analysis may also include determining the intensity, concentration, strength, or a combination thereof of the output brightness, which can be correlated with the amount of radiolabeled protein in the image, the area or region of the image, or a specific point on the image. Without being bound by theory, regions or points on an image that have greater intensity than other regions or points may have higher concentrations of radiolabeled proteins targeting, for example, amyloid deposits, and therefore may have higher concentrations of radiolabeled amyloid reactants or dyes attached to regions localized by amyloid reactants or dyes.

[0176] In some embodiments, methods for diagnosing the type of amyloidosis include analyzing images by the spatial location of the region of interest targeted by the applied amyloid reactant or detectable dye. In other embodiments, pharmacokinetic analysis of the applied amyloid reactant or dye can provide information about the appropriate timing of the injection. The presence or absence of amyloid can be determined by identifying areas, regions, or points on the image associated with the presence of the radiolabeled protein. For example, in some embodiments, identifying regions or points where the amyloid reactant or detectable dye is concentrated indicates the presence of amyloid. In some embodiments, images associated with the presence of the amyloid reactant are used to diagnose an individual's amyloidosis.

[0177] In some embodiments, the method further includes providing a diagnosis of the type of amyloidosis based on organ distribution patterns. In some embodiments, a specific organ distribution pattern indicates a specific type of amyloidosis. 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.

[0178] In some embodiments, different types of amyloidosis have different relative organ-to-organ ratios. In some embodiments, a particular type of amyloidosis may have a higher liver-to-heart ratio than another. In some embodiments, the heart-to-spleen ratio is higher in individuals diagnosed with AATR than in individuals diagnosed with AL. In some embodiments, the heart-to-spleen ratio is higher in individuals diagnosed with ATTR than in individuals diagnosed with ALECT2.

[0179] In some embodiments, the level of amyloid reactant in the heart is highest in individuals with ATTR. In some embodiments, the level of amyloid reactant in the liver is highest in individuals with AL. In some embodiments, the level of amyloid reactant is highest in the spleen of individuals with ALECT2. In some embodiments, the level of amyloid reactant is highest in the kidneys of individuals with ALECT2.

[0180] In some embodiments, amyloid responder levels are lowest in the heart of individuals with ALECT2. In some embodiments, amyloid responder levels are lowest in the liver of individuals with ATTR. In some embodiments, amyloid responder levels are lowest in the spleen of individuals with ATTR. In some embodiments, amyloid responder levels are lowest in the kidneys of individuals with ATTR.

[0181] In some implementations, the cutoff value for diagnosing a specific type of amyloidosis is selected based on a certain p-value. In some implementations, the cutoff is selected to provide a p-value less than 0.1, less than 0.05, less than 0.01, less than 0.005, or less than 0.001.

[0182] Those skilled in the art will understand that each ratio discussed herein can be readily 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).

[0183] In some embodiments, the diagnosis is confirmed by measures of an 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 diseases. In some embodiments, the diagnosis is confirmed by a cardiac biopsy. In some embodiments, the diagnosis is confirmed by additional amyloid imaging of the individual.

[0184] 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 the administration of the amyloid-beta reaction agent. In some embodiments, 130 mg of potassium iodide is administered for 3 days, starting on the same day as the administration of the amyloid-beta reaction agent, for example, about 30 minutes before the administration of the amyloid-beta reaction agent. D. Patient group

[0185] In some embodiments, the methods for diagnosing the type of amyloidosis are specifically designed for individuals at risk of developing amyloid-related diseases. In some embodiments, individuals at risk of developing amyloid-related diseases have a family history of amyloid-related diseases. In some embodiments, individuals are at risk of developing a genetic form of amyloid-related disease. In some embodiments, the genetic form of amyloid-related disease includes hereditary transthyretin amyloidosis (hATTR).

[0186] In some implementations, an individual's risk of amyloid-related disease is determined based on the presence of a gene mutation. In some implementations, the gene mutation is an ATTR gene mutation. In some implementations, the gene mutation is... TTR Within the gene. In some embodiments, gene mutations include point mutations that result in amino acid substitutions, such as the substitution of valine for methionine at position 30 (Val30 met).

[0187] In some implementations, an individual is identified as being at risk of developing amyloid-related disease based on having multiple myeloma. In some implementations, an individual at risk of developing amyloid-related disease has condensational multiple myeloma. In some implementations, an individual is at risk of developing immunoglobulin light chain amyloidosis (AI) based on having multiple myeloma.

[0188] In some implementations, an individual is identified as being at risk of amyloid-related disease based on having monoclonal gammopathy of undetermined significance (MGUS).

[0189] In some embodiments, an individual's risk of amyloid-related disease is determined based on laminectomy tissue that is amyloid-positive. In some embodiments, an individual's risk of amyloid-related disease is determined based on carpal tunnel release tissue that is amyloid-positive. In some embodiments, an individual's risk of amyloid-related disease is determined based on old age. 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.

[0190] 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: ankle and leg swelling, severe fatigue, shortness of breath, significant weight loss, difficulty swallowing, tingling, numbness or pain in the hands, wrists or feet, enlarged tongue, irregular heartbeat, diarrhea, and easily bruised, thickened or slightly purplish patches on the skin.

[0191] Alternatively, in some implementations, individuals at risk of developing amyloid-related disease do not exhibit symptoms of amyloid-related disease.

[0192] In some embodiments, an individual is suspected of having amyloid-related disease. In some embodiments, an individual is suspected of having amyloid-related disease but has not yet been diagnosed with it. 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, and 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, an individual suspected of having amyloid-related disease displays symptoms of amyloid-related disease. Alternatively, in some embodiments, the individual is at risk of developing amyloid-related disease and does not display symptoms of amyloid-related disease. In some embodiments, an individual suspected of having amyloid-related disease has a genetic predisposition to amyloid-related disease. In some embodiments, an individual suspected of having amyloid-related disease has a family history of any amyloid-related disease. In some embodiments, an individual suspected of having amyloid-related disease is an elderly person. In some embodiments, the individual has early-stage amyloid-related disease. E. Prognosis

[0193] This document also provides a method for determining the prognosis of an individual with amyloid-related disease, comprising: a) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated with any one of SEQ ID NO: 1-14, 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 first quantify amyloid in the individual; b) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated with any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to second quantify amyloid in the individual; and c) comparing the amyloid in the tissues or organs first quantified with the amyloid in the tissues or organs second quantified to determine the individual's prognosis.

[0194] In some embodiments, the individual has amyloidosis selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AIAPP, APro, Alns, APrP, or Aβ amyloidosis. In some embodiments, the individual has 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.

[0195] In some embodiments, the interval between the first and second applications of the amyloid-reactive peptide conjugated with the detectable label is at least one week, two weeks, three weeks, one month, two months, three months, four months, five months, six months, seven months, eight months, nine months, ten months, eleven months, one year, two years, three years, four years, or five years. In some embodiments, the interval between the first and second applications is at least two weeks. In some embodiments, the interval between the first and second applications is at least one month. In some embodiments, the interval between the first and second applications is at least three months.

[0196] In some implementations, the prognosis for individuals with amyloid-associated disease has worsened. In other implementations, the prognosis for individuals with amyloid-associated disease has improved.

[0197] In some embodiments, prognosis is confirmed by measures of an individual's health-related quality of life. 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. III. Treatment Methods

[0198] This article also provides a method for treating amyloid-related diseases, comprising: a) administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs; and c) administering treatment for amyloid-related diseases if the amount of the amyloid-reactive peptide is above a threshold.

[0199] This document provides a method for treating amyloid-related disease in an individual at an early stage, comprising administering an amyloid-reactive peptide conjugated to a detectable marker to the individual, detecting the amount of the detectable marker in the individual's tissues or organs, and administering treatment for said amyloid-related disease if the amount of the amyloid-reactive peptide is above a threshold. In some embodiments, an amount of amyloid-reactive peptide above a threshold indicates that the individual has amyloid-related disease, wherein the amyloid-related disease is in an early stage. 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 the detectable marker is a peptide containing... 124 A peptide of the amino acid represented by SEQ ID NO: 100 conjugated with I. In some embodiments, 124 I binds to amino acid 4 of SEQ ID NO: 100.

[0200] The methods provided herein can be used for the early treatment of amyloid-related diseases. In some embodiments, the methods can be used to treat amyloid-related diseases before the disease can be detected by conventional methods. In some embodiments, the individual has been previously diagnosed as not having amyloid-related diseases. In some embodiments, the methods have high sensitivity and specificity for the early detection and treatment of amyloid-related diseases. A. Application of amyloid reactive peptides

[0201] Some aspects of the present invention provide methods for treating amyloidosis based on the detection of amyloid protein using an amyloid protein reactant or a detection dye. In some embodiments, a method for treating amyloidosis is provided herein, comprising administering an amyloid protein reactant or a detection dye, measuring the organ distribution of the amyloid protein reactant or detection dye, and selecting treatment based on the type of disease.

[0202] In some embodiments, the method of treating amyloidosis includes administering an amyloid reactive agent or a detection dye, and measuring the organ-specific SUV and / or SUVR of the amyloid reactive agent for one or more organs. In some embodiments, the organ distribution pattern of the amyloid reactive agent or dye indicates the type of amyloidosis. In some embodiments, the method further includes selecting treatment based on the type of amyloidosis.

[0203] In some embodiments, the method includes obtaining an organ distribution pattern of an amyloid reactant or detection dye, wherein the organ distribution pattern indicates a specific type of amyloid disease, and administering treatment based on the amyloid disease.

[0204] In some embodiments, a method of treating amyloidosis includes administering an amyloid reactive agent comprising an amyloid reactive peptide conjugated to a detectable marker. In some embodiments, the amyloid reactive agent comprises a peptide, a fusion protein, a small molecule compound, or an antibody or fragment.

[0205] In some embodiments, a method of treating amyloidosis includes administering an amyloid reactive agent comprising an amyloid reactive peptide conjugated to a detectable marker. In some embodiments, the amyloid reactive peptide is a peptide having an amino acid sequence shown in any one of SEQ ID NO: 1-14. In some embodiments, a method of treating amyloidosis includes an amyloid reactive peptide having an amino acid sequence as shown in SEQ ID NO: 13.

[0206] In some embodiments, a method of treating amyloidosis includes administering an amyloid reactant or a detection dye, and measuring the organ-specific SUV and / or SUVR of the amyloid reactant for one or more organs. In some embodiments, a method of treating amyloidosis includes administering an amyloid reactant containing a detectable label to determine organ distribution patterns. In some embodiments, the amyloid reactant includes a fluorescent label, a chemiluminescent label, or a radioactive label. In some embodiments, the amyloid reactant includes a radioactive label. In some embodiments, the radioactively labeled amyloid reactant is... 124 I-p5+14. In other embodiments, the radiolabeled amyloid reactant is flubetapyr, flubetabenzene, and flumetazidine. In some embodiments, a method of treating amyloidosis includes p5+14. In some embodiments, the amyloid reactant is radiolabeled. In some embodiments, the radiolabeling is... 11 C 18 F, 111 In、 99m Tc and 123 I, 124 I or 125 I. In some embodiments, the radiolabeled amyloid reactive agent is a radiolabeled amyloid reactive peptide. In some embodiments, the radiolabeled amyloid reactive peptide is... 124 I-labeled amyloid reactive peptides. In other embodiments, methods for diagnosing the type of amyloidosis include administering... 124 I-p5+14. In other embodiments, the radiolabeled amyloid reactant is flubetapyr, flubetabenzene, or flumetazidine.

[0207] In some other embodiments, the amyloid reactant includes a fluorescent label. In some embodiments, the fluorescently labeled amyloid reactant is ThT. In some embodiments, the amyloid reactant is administered parenterally, adjacent to the cancer cell, transmucosally, percutaneously, intramuscularly, intravenously, intradermally, subcutaneously, intraperitoneally, intravenously, or intracranially. In some cases, the amyloid reactant is administered intravenously or intraperitoneally.

[0208] 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 the administration of the amyloid-beta reaction agent. In some embodiments, 130 mg of potassium iodide is administered for 3 days, starting on the same day as the administration of the amyloid-beta reaction agent, for example, about 30 minutes before the administration of the amyloid-beta reaction agent. B. Detection of amyloid protein levels

[0209] In some embodiments, a method of treating amyloidosis includes administering an amyloid reactive agent or a detection dye, and measuring the organ-specific SUV and / or SUVR of the amyloid reactive agent for one or more organs. In some embodiments, a method of treating amyloidosis includes measuring the organ distribution pattern of the amyloid reactive agent for one or more organs. In some embodiments, a method for treating said type of amyloidosis includes measuring the organ-specific SUV and / or SUVR of the amyloid reactive agent in one or more of the following organs: lung, fat, heart, kidney, pancreas, joint, spine, liver, spleen, adrenal gland, 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 kidney. In some embodiments, the step of measuring the organ distribution pattern of the amyloid reactive agent in one or more organs includes determining an organ uptake value for each organ. In some embodiments, the organ uptake value is a standard uptake value for each organ. In some embodiments, the standard uptake value is determined as the ratio of the amount of amyloid reactive agent detected in the organ to the amount of amyloid reactive agent detected in the blood. In some implementations, organ uptake values ​​indicate the organ distribution pattern of amyloid reactants or detection dyes.

[0210] In some embodiments, measuring the organ-specific SUV and / or SUVR of amyloid reactants includes analyzing imaging data generated by PET, CT, MRI, SPECT, PET / CT, PET / MRI, or other imaging techniques. In some embodiments, the step of measuring the organ-specific SUV and / or SUVR of amyloid reactants includes analyzing the images by spatial location of the region of interest.

[0211] In other embodiments, the step of measuring the organ distribution pattern of the amyloid reactive agent in one or more organs includes calculating organ-to-organ ratios for two or more organs. In some embodiments, calculating organ-to-organ ratios for two or more organs includes calculating the ratio between an organ uptake value for a first organ and an organ uptake value for a second organ. In some cases, the organ-to-organ ratio is selected from the group consisting of: liver and heart, spleen and heart, spleen and liver, spleen and kidney, kidney and heart, and kidney and liver. In some cases, the organ-to-organ ratio is the heart-to-spleen ratio. In some embodiments, the ratio is the reciprocal of any of these ratios.

[0212] In some embodiments, organ distribution patterns indicate the type of amyloidosis. In some embodiments, organ distribution patterns are used to select specific treatments based on the type of amyloidosis. In some embodiments, the method further includes providing a diagnosis of the type of amyloidosis based on organ distribution patterns. In some embodiments, a specific organ distribution pattern indicates a specific type of amyloidosis. 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. C. Application of treatment

[0213] In some embodiments, the method includes treating an individual with an amyloid-related disease or selecting a treatment for it. Some exemplary amyloid diseases that can be diagnosed and / or treated using the methods disclosed herein include, but are not limited to, type II diabetes, Alzheimer's disease, thyroid adenoma, Parkinson's disease, tau proteinosis, senile systemic amyloidosis, familial hemodialysis, senile systemic aging, senile pituitary disorders, iatrogenic syndromes, reactive chronic inflammation, thyroid adenoma, myeloma, or other forms of cancer with AA amyloidosis, AL amyloidosis, AH amyloidosis, Aβ amyloidosis, ATTR amyloidosis, ALECT2 amyloidosis, and IAPP amyloidosis. In some embodiments, the method of treating amyloid disease includes selecting a treatment for systemic amyloidosis. In some embodiments, the method of treating amyloid disease includes selecting a treatment for AL amyloidosis, ATTR amyloidosis, or ALECT2 amyloidosis. In some embodiments, the treatment is a targeted therapy for ATTR amyloidosis, AL amyloidosis, or ALECT2 amyloidosis. In some embodiments, the method of treating amyloidosis includes selecting a treatment for cardiac amyloidosis. In some embodiments, the method of treating amyloidosis includes selecting a treatment for cardiac AL amyloidosis. In some embodiments, the method of treating amyloidosis includes selecting a treatment for cardiac ATTR amyloidosis. In some embodiments, the amyloid-related disease is in an early stage. In some embodiments, the amyloid-related disease is early-stage amyloid-related disease.

[0214] In some embodiments, the method includes treating or selecting treatment for an individual suspected of having amyloid-related disease. In some embodiments, the individual is suspected of having amyloid-related disease but has not yet been diagnosed with it. In some embodiments, the individual has not undergone diagnostic evaluation for amyloid-related disease. In some embodiments, the individual has undergone diagnostic evaluation for amyloid-related disease, and 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 of having amyloid-related disease displays symptoms of amyloid-related disease. Alternatively, in some embodiments, the individual at risk of developing amyloid-related disease does not display 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, the individual suspected of having amyloid-related disease has a family history of any amyloid-related disease. In some embodiments, the individual suspected of having amyloid-related disease is an older person. In some embodiments, the individual has early-stage amyloid-related disease.

[0215] 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.

[0216] In some embodiments, the treatment is an antibody-peptide fusion protein comprising 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 NO: 1-14, comprising 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 shown in SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises: (i) an amyloid-reactive peptide comprising the amino acid sequence shown in SEQ ID NO: 2; and (ii) an antibody that binds to human amyloid fibrils.

[0217] In some embodiments, the antibody binding to human amyloid fibrils comprises a heavy chain and a light chain. In some embodiments, the antibody binding to human amyloid fibrils comprises a heavy chain variable region (VH), and the light chain of the antibody comprises a light chain variable region (VL), as shown in Table 2. In some embodiments, the VH comprises the amino acid shown in SEQ ID NO: 102. In some embodiments, the VL comprises the amino acid shown in SEQ ID NO: 101. Table 2: Exemplary antibodies VH and VL

[0218] 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), wherein the VH and VV contain 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), wherein the VH contains: CDR-H1 containing the amino acid sequence shown in SEQ ID NO: 106, CDR-H2 containing the amino acid sequence shown in SEQ ID NO: 107, and CDR-H3 containing the amino acid sequence shown in SEQ ID NO: 108, and the VL contains: CDR-L1 containing the amino acid sequence shown in SEQ ID NO: 103, CDR-L2 containing the amino acid sequence shown in SEQ ID NO: 104, and CDR-L3 containing the amino acid sequence shown in SEQ ID NO: 105. Table 3: Exemplary Antibody CDRs

[0219] In some embodiments, 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 VSPSV (SEQ ID NO: 83). In some embodiments, the antibody-peptide fusion may be any modified immunoglobulin described in U.S. Patent Publication 12,030,934B2.

[0220] 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 includes 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 includes an antibody or fragment that binds to misfolded TTRs. In some embodiments, the antibody is PRX004. In some embodiments, the treatment includes an oligonucleotide. In some embodiments, the oligonucleotide is a TTR silencer. In some embodiments, the TTR silencer is Partisan (ALN-TTR02), uterisan, inostatin, or AKCEA-TTR-LRx. In some embodiments, the treatment includes an ATTR amyloid disruptor. In some embodiments, the treatment includes doxycycline, tauroursodeoxycholic acid, or serum amyloid P (SAP). In some implementations, the treatment includes organ transplantation. In some implementations, the treatment includes liver transplantation.

[0221] In other embodiments, the treatment is a targeted therapy for AL amyloidosis. In some embodiments, the treatment includes bortezomib, ixazomib, or carfilzomib. In some embodiments, the treatment includes an antibody or fragment. In some embodiments, the treatment includes daratumumab, CAEL-101, elotuzumab, or belantamab mafodotin. In some embodiments, the treatment includes stem cell therapy. In some embodiments, the treatment includes corticosteroids. In some embodiments, the corticosteroid is dexamethasone.

[0222] In some embodiments, the method is used to eliminate potential treatments for patients with amyloidosis. In some embodiments, the method is used to diagnose one type of amyloidosis and exclude treatments for other types of amyloidosis. In some embodiments, the method is used to diagnose ALECT2 and exclude therapies for AL or ATTR amyloidosis.

[0223] In some embodiments, the method is used to differentiate between 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.

[0224] In some embodiments, this document provides a method for monitoring treatment of an individual for amyloid-related disease, comprising: a) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated with any one of SEQ ID NO: 1-14, and detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable label at the individual's tissues or organs to first quantify amyloid in the individual; b) administering treatment for amyloid-related disease; c) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated with any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label at the individual's tissues or organs to second quantify amyloid in the individual; and d) comparing the first quantification of amyloid in the tissues or organs with the second quantification of amyloid in the tissues or organs to determine whether the treatment is effective.

[0225] In some embodiments, the interval between the first and second administrations of the amyloid-reactive peptide is at least one week, two weeks, three weeks, one month, two months, three months, four months, five months, six months, seven months, eight months, nine months, ten months, eleven months, one year, two years, three years, four years, or five years. In some embodiments, the interval between the first and second administrations is at least two weeks. In some embodiments, the interval between the first and second administrations is at least one month. In some embodiments, the interval between the first and second administrations is at least three months.

[0226] In some embodiments, this document provides a method for selecting a treatment for an amyloid-associated disease in an individual, comprising: a) administering an amyloid-reactive peptide to the individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and b) detecting the amount of the amyloid-reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs, wherein if amyloid is detected in the heart, treatment for the amyloid-associated disease is administered, and wherein if amyloid is not detected in the heart, an alternative therapy is administered. IV. Reagent Kit

[0227] Some aspects of the present invention provide kits for diagnosing or detecting the type of amyloid disease in an individual using the methods described herein.

[0228] In some embodiments, the kit includes an amyloid reactant and instructions for use. In some embodiments, the amyloid reactant contains a detectable marker. In some embodiments, the amyloid reactant is... 124I-p5+14. In some embodiments, the amyloid reactant comprises an amino acid sequence containing SEQ ID NO: 13.

[0229] In some implementations, the radioactive label is 11 C 18 F, 111 In、 99m Tc, 123 I, 124 I or 125 I. In some embodiments, the radiolabeled amyloid reactive agent is a radiolabeled amyloid reactive peptide. In some embodiments, the radiolabeled amyloid reactive peptide is... 124 I-labeled amyloid reactive peptides. In other embodiments, methods for diagnosing the type of amyloidosis include administering... 124 I-p5+14. In other embodiments, the radiolabeled amyloid reactant is flubetapyr, flubetabenzene, or flumetazidine.

[0230] In some embodiments, the instructions include instructions for using an amyloid reactant to detect amyloid in one or more organs. In some embodiments, the amyloid reactant is detected in blood, heart, lung, kidney, or spleen.

[0231] In some embodiments, the kit includes instructions for calculating organ-specific SUVs. In some embodiments, the kit includes instructions for calculating organ-specific SUVRs. In some embodiments, the kit includes instructions for calculating the SUVR ratio for one or more organs. In some embodiments, a blood pool is used as a reference tissue to calculate the SUVR. In some embodiments, the SUVR for each organ is calculated by dividing the amount of amyloid detection agent 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.

[0232] In some embodiments, the specification includes instructions for determining organ-to-organ ratios. In some embodiments, the organ-to-organ ratio is liver to heart, spleen to heart, spleen to liver, spleen to kidney, kidney to heart, kidney to liver, or any reciprocal of these ratios.

[0233] In some embodiments, the kit also 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 organ-to-organ ratios.

[0234] In some implementations, the kit includes a therapeutic agent for treating amyloidosis. Exemplary implementation 1. A method for diagnosing amyloid-related disease in individuals at risk of developing the disease, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b. The amount of amyloid reactive peptides can be detected by measuring the amount of a detectable marker in an individual's tissues or organs. The presence of amyloid reactive peptides above the threshold indicates that an individual has amyloidosis. 2. The method of Implementation 1, wherein an individual is identified as being 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 from carpal tunnel release surgery, having monoclonal gammopathy of undetermined 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 older. 3. The method of embodiment 2, wherein the gene mutation is in transthyretin. 4. The method of embodiment 2, wherein the gene mutation is in fibrinogen α protein. 5. The method of any one of embodiments 1-3, wherein the individual does not have symptoms of amyloidosis. 6. The method of any one of embodiments 1-3, wherein the individual has neuropathological symptoms of amyloidosis. 7. A method for diagnosing amyloid-associated disease in an individual suspected of having such disease, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b. The amount of amyloid reactive peptides can be detected by measuring the amount of a detectable marker in an individual's tissues or organs. The presence of amyloid reactive peptides above the threshold indicates that an individual has amyloidosis. 8. A method for determining the prognosis of an individual with amyloid-related disease, comprising: a. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 for the first time; b. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to a second quantification of amyloid in the individual; and c. Compare the amyloid protein quantified for the first time in a tissue or organ with the amyloid protein quantified for the second time in a tissue or organ to determine the individual's prognosis. 9. The method of embodiment 8, wherein the prognosis of an individual with amyloid-related disease is based on the individual detection of the amount of amyloid reactive peptide in the heart. 10. The method of embodiment 8, wherein the prognosis of an individual with amyloid-related disease is based on the detection of the amount of amyloid reactive peptides in the heart and kidneys. 11. The method of embodiment 8, wherein the prognosis of an individual suffering from amyloid-related disease is based on detecting the amount of amyloid reactive peptides in the heart, kidneys, and all other organs. 12. A method for treating amyloid-related diseases, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence shown in any one of SEQ ID NO: 1-14 conjugated to a detectable marker; b. Detecting the amount of amyloid reactive peptides by measuring the amount of detectable markers in an individual's tissues or organs, and c. If the amount of amyloid reactive peptides is above the threshold, it is used for the treatment of amyloid-related diseases. 13. A method for monitoring the treatment of amyloid-related diseases in an individual, comprising: a. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 for the first time; b. For the treatment of amyloid-related diseases; c. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to a second quantification of amyloid in the individual; and d. Compare the amyloid protein measured for the first time in a tissue or organ with the amyloid protein measured for the second time in the tissue or organ to determine whether the treatment is effective. 14. The method of any one of embodiments 1-13, further comprising administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 a second, optionally a third, fourth and / or fifth time. 15. The method of embodiment 13 or 14, wherein the first and second times are spaced at least six weeks apart. 16. A method for selecting individuals for the treatment of amyloid-related diseases, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b. The amount of amyloid reactive peptides can be detected by measuring the amount of a detectable marker in an individual's tissues or organs. If amyloid protein is detected in the heart, treatment for amyloid-related diseases is administered; if amyloid protein is not detected in the heart, replacement therapy is administered. 17. A method for managing the treatment of amyloid-related diseases in an individual, comprising: a. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 for the first time; b. For the treatment of amyloid-related diseases; c. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to a second quantification of amyloid in the individual; and d. Compare the amyloid protein quantified for the first time in a tissue or organ with the amyloid protein quantified for the second time in the tissue or organ. e. Adjust the treatment used for amyloid-related diseases. 18. The method of embodiment 17, wherein if the amyloidosis is stable between the first and second treatments, the treatment is changed. 19. The method of embodiment 18, wherein if the amyloid load increases, treatment is restarted. 20. The method of any one of embodiments 1-19, wherein the amyloid reactive peptide comprises SEQ ID NO: 13. 21. The method of any one of embodiments 1-20, wherein the detectable marker is a radioactive marker. 22. The method of any one of embodiments 1-21, wherein the detectable marker is selected from... 11 C 18 F, 111 In、 99m Tc, 89 Zr、 68 Ga and 123 I, 124 I and 125 Group I. 23. The method of any one of embodiments 1-22, wherein the detectable marker is 124 I. 24. The method of any one of embodiments 1-23, comprising administering to an individual a dose of about 0.1 to about 25 mCi. 124 I-labeled amyloid reactive peptide. 25. The method of any one of embodiments 1-24, comprising administering to an individual a dose of about 0.3 mCi, 1 mCi, or about 2 mCi. 124 I-labeled amyloid reactive peptide. 26. The method of any one of embodiments 1-25, comprising administering Tc-99 to an individual at a dose of about 20 mCi. 27. The method of any one of embodiments 1-26, comprising administering to an individual about 0.3 mg, 1 mg, 1.5 mg or about 2 mg of amyloid reactive peptide. 28. The method of any one of embodiments 1-27, wherein the amyloid reactive peptide is administered intravenously. 29. The method of any one of embodiments 1-28 further includes determining the organ-specific standard uptake ratio for an individual's internal organs. 30. The method of embodiment 29, wherein the organ-specific SUVR for an individual is selected from the group consisting of the SUVR average, the SUVR maximum, and the SUVR peak. 31. The method of any one of embodiments 1-30, further comprising determining an organ-specific SUV for an individual. 32. The method of embodiment 31, wherein the organ-specific SUV for an individual is selected from the group consisting of SUV average, SUV maximum and SUV peak. 33. The method of any one of embodiments 1-32, further comprising determining the organ-specific percentage injection dose / gram (%ID) for an individual. 34. The method of any one of embodiments 1-33, wherein the amyloid reactive peptide has a sensitivity of at least 90%. 35. The method of any one of embodiments 1-34, wherein the sensitivity is about 99% or about 100%. 36. The method of any one of embodiments 1-35, further comprising determining one or more health-related quality of life measures for an individual. 37. The method of any one of embodiments 1-36, further comprising detecting one or more biomarkers associated with amyloid-related diseases. 38. The method of embodiment 37, wherein the biomarkers associated with amyloid-related diseases are selected from the group consisting of troponin T, NTproBNP, urinary protein levels, UACR, EGFR and alkaline phosphatase levels. 39. The method of any one of embodiments 1-38, wherein the tissues or organs of the individual are selected from the group consisting of 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. 40. The method of any one of embodiments 1-39, further comprising performing a cardiac biopsy if amyloid protein is detected in the heart. 41. The method of any one of embodiments 1-40, further comprising performing additional amyloid imaging on the individual. 42. The method of embodiment 41, wherein additional amyloid imaging of the individual includes ECHO, CMR, bone scintillation scanning, or positron emission tomography. 43. The method of embodiment 41, wherein further imaging of an individual's amyloid protein includes a tracer selected from the group consisting of: 99m Tc-PyP, 99m Tc-DPD, 99mTc-HMDP, 99m Tc-MDP, other bone scintillation tracers 124 I and 18 F-Fluorbetapyr 18 F-Fluorometabolone and 18 F-Fluorobetabenzene. 44. The method of any one of embodiments 1-43, wherein a diagnosis, prognosis or response is confirmed by measures of an individual’s health-related quality of life, the presence or amount of one or more biomarkers associated with amyloid-related diseases, cardiac biopsy and / or additional amyloid imaging of the individual. 45. The method of any one of embodiments 1-44, wherein the individual is diagnosed with amyloid cardiomyopathy. 46. ​​The method of any one of embodiments 1-45, wherein the individual is suspected of having amyloid cardiomyopathy. 47. The method of any one of embodiments 1-46, wherein the amyloid-related disease is systemic or localized amyloidosis. 48. The method of any one of embodiments 1-47, wherein the amyloid reactive peptide has pan-amyloid specificity. 49. The method of any one of embodiments 1-48, wherein the amyloid reactive peptide binds to the following amyloid proteins: 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), gellingin (AGel), apolipoprotein C-II (AApoCII), and apolipoprotein C-I. I (AApoCIII), lysozyme (ALys), leukocyte chemotactic factor (ALECT2), fibrinogen a variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactoglobulin (AMed), islet amyloid peptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP); α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), IAAP, ALβ4 or ALβ1. 50. The method of any one of embodiments 1-49, wherein PET, PET / CT, gamma scintillation scanning, SPECT and / or SPECT / CT are used to detect amyloid reactive peptides. 51. The method of any one of embodiments 1-50, wherein amyloid-related diseases are selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AApoCII, AApoCIII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AIAPP, APro, AIns, APrP and Aβ amyloidosis. 52. The method of any one of embodiments 1-51, wherein the individual has a genetic predisposition to amyloid-related diseases. 53. The method of any one of embodiments 1-52, wherein the individual has a family history of amyloid-related diseases. 54. The method of any one of embodiments 1-53, wherein the individual is an elderly person. 55. The method of any one of embodiments 1-54, wherein the individual suffers from early amyloid-related disease. 56. The method of any one of embodiments 1-55, wherein the individual has early AL amyloidosis. 57. The method of embodiment 56, wherein early AL amyloidosis is diagnosed according to the Mayo clinical system. 58. The method of embodiment 57, wherein the early AL amyloidosis is stage 1 AL amyloidosis. 59. The method of any one of embodiments 1-58, wherein the individual has early ATTR amyloidosis. 60. The method of embodiment 59, wherein early ATTR amyloidosis includes stage 1 ATTR amyloidosis. 61. The method of any one of embodiments 1-11, 14-16 and 18-60, further comprising administering a therapy for amyloid-related diseases. 62. The method of any one of embodiments 12-13, 17 and 61, wherein the therapy for amyloid-related diseases is selected from the group consisting of: transthyretin stabilizers (e.g., tafamidix, aclamidix), transthyretin silencers (e.g., Partisan, Inothesen, Vutrisan, Eploetersen), gene editing methods, anti-amyloid methods using monoclonal antibodies, therapy targeting plasma cell clones (e.g., daratumumab, bortezomib), and antibody-peptide fusions comprising an antibody-peptide fusion protein comprising a second amyloid reactive peptide and an antibody binding to amyloid fibrils. 63. The method of embodiment 62, wherein the second amyloid reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-14 containing 0, 1, 2, 3 or 4 amino acid substitutions, insertions or deletions. 64. The method of embodiment 62 or 63, wherein the antibody-peptide fusion protein comprises a heavy chain and a light chain, and wherein the amyloid reactive peptide is linked to the C-terminus of the light chain of the antibody via a spacer. 65. The method of any one of embodiments 62-64, wherein the antibody-peptide fusion protein comprises: (i) an amyloid reactive peptide comprising the amino acid sequence shown in SEQ ID NO: 2; and (ii) An antibody that binds to human amyloid fibrils, 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 the VH comprises: CDR-H1 comprising the amino acid sequence shown in SEQ ID NO: 106, CDR-H2 comprising the amino acid sequence shown in SEQ ID NO: 107 and CDR-H3 comprising the amino acid sequence shown in SEQ ID NO: 108, and the VL comprises: CDR-L1 comprising the amino acid sequence shown in SEQ ID NO: 103, CDR-L2 comprising the amino acid sequence shown in SEQ ID NO: 104 and CDR-L3 comprising the amino acid sequence shown in SEQ ID NO: 105; wherein the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain, and wherein the amyloid reactive peptide is linked to the antibody via a spacer comprising the amino acid sequence shown in SEQ ID NO: 83. 66. The method of any one of embodiments 1-65, wherein an individual is given a peptide comprising an amyloid-reactive peptide and an N-terminal leader sequence. 67. The method of embodiment 66, wherein the peptide comprising the amyloid reactive peptide comprises the amino acid sequence shown in SEQ ID NO: 100. 68. The method of embodiment 67, wherein the detectable tag binds to one or more amino acids at positions 1 to 6 of the amino acid sequence shown in SEQ ID NO: 100. 69. The method of embodiment 68, wherein the detectable marker is 124 I. 70. The method of embodiment 69, wherein... 124 I binds to amino acid 4 of SEQ ID NO: 100. 71. A peptide comprising the amino acids shown in SEQ ID NO: 100, wherein the peptide comprises a detectable tag binding to one or more amino acids at positions 1 to 6. 72. The peptide of embodiment 71, wherein the detectable marker is 124 I. 73. The peptide described in embodiment 72, wherein... 124 I binds to amino acid 4 of SEQ ID NO: 100. 1A. A method for early diagnosis of amyloid-related disease in individuals at risk of developing the disease, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b. The amount of amyloid reactive peptides can be detected by measuring the amount of a detectable marker in an individual's tissues or organs. An amount of amyloid reactive peptides above the threshold indicates that an individual has amyloid-related disease, which is in the early stage of amyloid-related disease. 2A. The method of Implementation 1A, wherein an individual is identified as being 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 from carpal tunnel release surgery, having monoclonal gammopathy of undetermined 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 older. 3A. The method of embodiment 2A, wherein the gene mutation is in transthyretin. 4A. The method of embodiment 2A, wherein the gene mutation is in fibrinogen α protein. 5A. The method of any one of embodiments 1A-3A, wherein the individual does not have symptoms of amyloidosis. 6A. The method of any one of embodiments 1A-3A, wherein the individual has neuropathological symptoms of amyloidosis. 7A. The method of any one of embodiments 1A-6A, wherein the individual has been previously diagnosed as not having amyloid-related disease. 8A. A method for early diagnosis of amyloid-related disease in individuals suspected of having it, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b. The amount of amyloid reactive peptides can be detected by measuring the amount of a detectable marker in an individual's tissues or organs. An amount of amyloid reactive peptides above the threshold indicates that an individual has amyloidosis, with amyloid-associated disease being in its early stages. 9A. A method for determining the prognosis of an individual with early-stage amyloid-related disease, comprising: a. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 for the first time; b. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to a second quantification of amyloid in the individual; and c. The amyloid protein quantification in a tissue or organ for the first time will be compared with the amyloid protein quantification in a tissue or organ for the second time to determine the individual's prognosis, where amyloid-associated disease is in its early stages. 10A. The method of Implementation 9A, wherein the prognosis of an individual with amyloid-related disease is based on the individual detection of the amount of amyloid reactive peptide in the heart. 11A. The method of embodiment 9A, wherein the prognosis of an individual with amyloid-related disease is based on the detection of the amount of amyloid reactive peptides in the heart and kidneys. 12A. The method of embodiment 9A, wherein the prognosis of an individual with amyloid-related disease is based on the detection of the amount of amyloid reactive peptides in the heart, kidneys and all other organs. 13A. A method for treating amyloid-associated disease in an individual with early-stage amyloid-associated disease, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence shown in any one of SEQ ID NO: 1-14 conjugated to a detectable marker; b. Detecting the amount of amyloid reactive peptides by measuring the amount of detectable markers in an individual's tissues or organs, and c. If the amount of amyloid reactive peptides is above the threshold, it is used for the treatment of amyloid-related diseases. 14A. The method of implementation 13A, wherein the individual is suspected of having amyloid-related disease. 15A. A method for monitoring the treatment of amyloid-related disease in individuals with early-stage amyloid-related disease, comprising: a. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 for the first time; b. For the treatment of amyloid-related diseases; c. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to a second quantification of amyloid in the individual; and d. Compare the amyloid protein measured for the first time in a tissue or organ with the amyloid protein measured for the second time in the tissue or organ to determine whether the treatment is effective. 16A. The method of any one of embodiments 1A-15A, further comprising administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 a second, optionally a third, fourth and / or fifth time. 17A. The method described in implementation 15A or 16A, wherein the first and second times are spaced at least six weeks apart. 18A. A method for selecting individuals for the treatment of amyloid-related diseases, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b. The amount of amyloid reactive peptides can be detected by measuring the amount of a detectable marker in an individual's tissues or organs. If amyloid protein is detected in the heart, treatment for amyloid-related diseases is administered; if amyloid protein is not detected in the heart, replacement therapy is administered. 19A. A method for managing the treatment of amyloid-related diseases in an individual, comprising: a. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 for the first time; b. For the treatment of amyloid-related diseases; c. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to a second quantification of amyloid in the individual; and d. Compare the amyloid protein quantified for the first time in a tissue or organ with the amyloid protein quantified for the second time in the tissue or organ. e. Adjust the treatment used for amyloid-related diseases. 20A. The method of embodiment 19A, wherein if the amyloidosis is stable between the first and second treatments, the treatment is changed. 21A. The method of embodiment 20A, wherein if the amyloid load increases, treatment is restarted. 22A. The method of embodiments 9A-12A and 14A-21A, wherein an amount of amyloid reactive peptide above a threshold indicates that an individual has an amyloid-related disease. 23A. The method of embodiment 22A, wherein the amyloid-associated disease is in the early stage of amyloid-associated disease. 24A. The method of any one of embodiments 1A-23A, wherein the amyloid reactive peptide comprises SEQ ID NO: 13. 25A. The method of any one of embodiments 1A-24A, wherein the detectable marker is a radioactive marker. 26A. The method of any one of embodiments 1A-25A, wherein the detectable marker is selected from... 11 C 18 F, 111 In、 99m Tc, 89 Zr、 68 Ga and 123 I, 124 I and 125 Group I. 27A. The method of any one of embodiments 1A-26A, wherein the detectable mark is124 I. 28A. The method of any one of embodiments 1A-27A, comprising administering to an individual a dose of about 0.1 to about 25 mCi. 124 I-labeled amyloid reactive peptide. 29A. The method of any one of embodiments 1A-28A, comprising administering to an individual a dose of about 0.3 mCi, 1 mCi, or about 2 mCi. 124 I-labeled amyloid reactive peptide. 30A. The method of any one of embodiments 1A-29A, comprising administering to an individual a dose of about 1 mCi (±10%). 124 I-labeled amyloid reactive peptide. 31A. The method of any one of embodiments 1A-30A, comprising administering Tc-99 to an individual at a dose of about 20 mCi. 32A. The method of any one of embodiments 1A-31A, comprising administering to an individual about 0.3 mg, 1 mg, 1.5 mg or about 2 mg of an amyloid reactive peptide. 33A. The method of any one of embodiments 1A-23A, wherein the amyloid reactive peptide is administered intravenously. 34A. The method of any one of embodiments 1A-33A, further comprising determining the organ-specific standard uptake ratio for an individual's internal organs. 35A. The method of embodiment 34A, wherein the organ-specific SUVR for an individual is selected from the group consisting of SUVR average, SUVR maximum, and SUVR peak. 36A. The method of any one of embodiments 1A-35A, further comprising determining an organ-specific SUV for an individual. 37A. The method of embodiment 34A, wherein the organ-specific SUV for an individual is selected from the group consisting of SUV average, SUV maximum, and SUV peak. 38A. The method of any one of embodiments 1A-37A, further comprising determining the organ-specific percentage injection dose / gram (%ID) for an individual. 39A. The method of any one of embodiments 1A-38A, wherein the sensitivity of the amyloid reactive peptide for detecting amyloid 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%. 40A. The method of any one of embodiments 1A-39A, wherein the sensitivity is about 99% or about 100%. 41A. The method of any one of embodiments 1A-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%. 42A. The method of any one of embodiments 1A-41A, wherein the specificity of the amyloid reactive peptide is about 99% or 100%. 43A. The method of any one of embodiments 1A-42A, further comprising determining one or more health-related quality of life measures for an individual. 44A. The method of any one of embodiments 1A-43A, further comprising detecting one or more biomarkers associated with amyloid-related diseases. 45A. The method of embodiment 44A, wherein the biomarkers associated with amyloid-related diseases are selected from the group consisting of troponin T, NTproBNP, urinary protein levels, UACR, EGFR and alkaline phosphatase levels. 46A. The method of any one of embodiments 1A-45A, wherein the tissues or organs of the individual are selected from the group consisting of 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. 47A. The method described in embodiments 1A-8A, 13A and 22A-46A, wherein the threshold is the SUVR average threshold. 48A. The method of embodiment 47A, wherein if the organ or tissue is the liver, the average 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 average SUVR threshold is approximately 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5; and if the organ or tissue is the kidney, the average SUVR threshold is approximately 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, or 2.3. If the organ or tissue is the pancreas, the average 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 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. 49A. The method of embodiment 48A, wherein the average SUVR threshold is 1.31 if the organ or tissue is the liver, 1.21 if the organ or tissue is the spleen, 1.88 if the organ or tissue is the kidney, 1.40 if the organ or tissue is the pancreas, 1.19 if the organ or tissue is the heart, or 1.35 if the organ or tissue is the adrenal gland. 50A. The method described in embodiments 1A-8A, 13A and 22A-46A, wherein the threshold is the maximum threshold of SUVR. 51A. The method of embodiment 50A, wherein 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, or 10. The maximum SUVR thresholds are 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. 52A. The method of embodiment 51A, wherein the maximum SUVR threshold is 13.26 if the organ or tissue is the liver, 4.86 if the organ or tissue is the spleen, 9.68 if the organ or tissue is the kidney, 9.48 if the organ or tissue is the pancreas, 6.20 if the organ or tissue is the heart, or 1.31 if the organ or tissue is the adrenal gland. 53A. The method of any one of embodiments 1A-52A, further comprising performing a cardiac biopsy if amyloid protein is detected in the heart. 54A. The method of any one of embodiments 1A-53A, further comprising performing additional amyloid imaging on the individual. 55A. The method of any one of embodiments 1A-54A, further comprising performing additional amyloid imaging of the individual prior to the application of the amyloid reactive peptide. 56A. The method of embodiment 55A, wherein additional amyloid imaging of the individual includes ECHO, CMR, bone scintillation or positron emission tomography. 57A. The method of embodiment 56A, wherein further imaging of amyloid in an individual includes a tracer selected from the group consisting of: 99m Tc-PyP, 99m Tc-DPD, 99m Tc-HMDP, 99m Tc-MDP, other bone scintillation tracers 124 I and 18 F-Fluorbetapyr 18 F-Fluorometabolone and 18 F-Fluorobetabenzene. 58A. The method of any one of embodiments 1A-57A, wherein a diagnosis, prognosis or response is confirmed by measures of an individual’s health-related quality of life, the presence or amount of one or more biomarkers associated with amyloid-related diseases, cardiac biopsy and / or additional amyloid imaging of the individual. 59A. The method of any one of embodiments 1A-58A, wherein the individual is diagnosed with amyloid cardiomyopathy. 60A. The method of any one of embodiments 1A-59A, wherein the individual is suspected of having amyloid cardiomyopathy. 61A. The method of any one of embodiments 1A-60A, wherein the amyloid-associated disease is systemic or localized amyloidosis. 62A. The method of any one of embodiments 1A-61AA, wherein the amyloid-related disease is cardiac amyloidosis. 63A. The method of any one of embodiments 1A-62A, wherein the amyloid reactive peptide has pan-amyloid specificity. 64A. The method of any one of embodiments 1A-63A, wherein the amyloid reactive peptide binds to the following amyloid proteins: 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), colloidin (AGel), apolipoprotein C-II (AApoCII), apolipoprotein C- II (AApoCIII), lysozyme (ALys), leukocyte chemotactic factor (ALECT2), fibrinogen a variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactoglobulin (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. 65A. The method of any one of embodiments 1A-64A, wherein amyloid reactive peptides are detected using PET, PET / CT, gamma scintillation scanning, SPECT and / or SPECT / CT. 66A. The method of any one of embodiments 1A-65A, wherein amyloid-related diseases are selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AApoCII, AApoCIII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AIAPP, APro, AIns, APrP and Aβ amyloidosis. 67A. The method of embodiment 66A, wherein the amyloid-related disease is cardiac AL amyloidosis. 68A. The method of embodiment 66A, wherein the amyloid-related disease is cardiac ATTR amyloidosis. 69A. The method of any one of embodiments 1A-68A, wherein the individual has a genetic predisposition to amyloid-related diseases. 70A. The method of any one of embodiments 1A-69A, wherein the individual has a family history of amyloid-related diseases. 71A. The method of any one of embodiments 1A-70A, wherein the individual is an elderly person. 72A. The method of any one of embodiments 1A-71A, wherein the individual suffers from early amyloid-related disease. 73A. The method of any one of embodiments 1A-72A, wherein the individual suffers from early systemic amyloidosis. 74A. The method of any one of embodiments 1A-73A, wherein the individual suffers from early cardiac amyloidosis. 75A. The method of any one of embodiments 1A-74A, wherein the individual has early AL amyloidosis. 76A. The method of embodiment 75A, wherein early AL amyloidosis is diagnosed according to the Mayo clinical system. 77A. The method of embodiment 76A, wherein the early AL amyloidosis is stage 1 AL amyloidosis. 78A. The method of any one of embodiments 1A-77A, wherein the individual has early ATTR amyloidosis. 79A. The method of embodiment 78A, wherein early ATTR amyloidosis includes stage 1 ATTR amyloidosis. 80A. The method of any one of embodiments 1A-12A, 16A-18A and 20A-79A, further comprising administration for treatment of amyloid-related diseases. 81A. The method of any one of embodiments 13A-15A, 19A and 80A, wherein the treatment for amyloid-related diseases is selected from the group consisting of: transthyretin stabilizers (e.g., tafamidix, aclamidix), transthyretin silencers (e.g., Partisan, Inothesen, Vutrisan, Eploetersen), gene editing methods, anti-amyloid methods using monoclonal antibodies, treatments targeting plasma cell clones (e.g., daratumumab, bortezomib), and antibody-peptide fusions comprising an antibody-peptide fusion protein comprising a second amyloid reactive peptide and an antibody binding to amyloid fibrils. 82A. The method of embodiment 81A, wherein the second amyloid reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-14 containing 0, 1, 2, 3 or 4 amino acid substitutions, insertions or deletions. 83A. The method of embodiment 81A or 82A, wherein the antibody-peptide fusion protein comprises a heavy chain and a light chain, and wherein the amyloid reactive peptide is linked to the C-terminus of the light chain of the antibody via a spacer. 84A. The method of any one of embodiments 81A-83A, wherein the antibody-peptide fusion protein comprises: (i) an amyloid reactive peptide comprising the amino acid sequence shown in SEQ ID NO: 2; and (ii) An antibody that binds to human amyloid fibrils, 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 the VH comprises: CDR-H1 comprising the amino acid sequence shown in SEQ ID NO: 106, CDR-H2 comprising the amino acid sequence shown in SEQ ID NO: 107 and CDR-H3 comprising the amino acid sequence shown in SEQ ID NO: 108, and the VL comprises: CDR-L1 comprising the amino acid sequence shown in SEQ ID NO: 103, CDR-L2 comprising the amino acid sequence shown in SEQ ID NO: 104 and CDR-L3 comprising the amino acid sequence shown in SEQ ID NO: 105; wherein the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain, and wherein the amyloid reactive peptide is linked to the antibody via a spacer comprising the amino acid sequence shown in SEQ ID NO: 83. 85A. The method of any one of embodiments 1A-84A, wherein an individual is given a peptide comprising an amyloid-reactive peptide and an N-terminal leader sequence. 86A. The method of embodiment 85A, wherein the peptide comprising the amyloid reactive peptide comprises the amino acid sequence shown in SEQ ID NO: 100. 87A. The method of embodiment 86A, wherein the detectable marker binds to one or more amino acids at positions 1 to 6 of the amino acid sequence shown in SEQ ID NO: 100. 88A. The method of embodiment 87A, wherein the detectable marker is 124 I. 89A. The method of embodiment 88A, wherein 124 I binds to amino acid 4 of SEQ ID NO: 100. 90A. The method of any one of embodiments 1A-89A, further comprising administering potassium iodide to an individual. 91A. The method of embodiment 90A, wherein the administration of potassium iodide comprises administering 130 mg of potassium iodide to the patient for seven days, starting one day prior to the administration of the amyloid reactive peptide. 92A. The method of embodiment 91A, wherein the administration of potassium iodide comprises administering 130 mg of potassium iodide to the patient for three days, starting approximately 30 minutes prior to the administration of the amyloid reactive peptide. 93A. The method of any one of embodiments 90A-92A, wherein the potassium iodide is administered orally. 94A. A peptide comprising the amino acid shown in SEQ ID NO: 100, wherein the peptide comprises a detectable tag that binds to one or more amino acids at positions 1 to 6. 95A. The peptide of embodiment 94A, wherein the detectable marker is 124 I. 96A. The peptide of embodiment 95A, wherein 124 I binds to amino acid 4 of SEQ ID NO: 100. Example

[0235] The following examples are for illustrative purposes only and are not intended to limit the scope of this disclosure.

[0236] The scope of this disclosure is not limited to the specific embodiments described herein, which are provided to illustrate various aspects of this disclosure. Various modifications to the described compositions and methods will become apparent from the description and teachings herein. Such changes may be practiced without departing from the true scope and spirit of this disclosure and are intended to fall within its scope. Example 1: By 124 Cardiac amyloid detection on PET / CT imaging at I-p5+14

[0237] This example describes the safety and efficacy of a novel amyloid reactive peptide labeled with iodine-124 (named p5+14) in patients with different types of systemic amyloidosis. Research Design

[0238] This is a multipart, open-label study to evaluate 124 Safety, dosimetry, and biodistribution of the I-p5+14 peptide in patients with any type of systemic amyloidosis; presymptomatic subjects carrying germline TTR mutations; and healthy subjects. In Part 5, repeat imaging was performed on individual patients at approximately two-year intervals (Table 5). Patients with systemic AL amyloidosis were initially enrolled to evaluate radiation dosimetry, followed by additional patients. Inclusion targets were not met due to premature termination of the study to allow the transfer of the IND to a commercial partner for clinical development (Table 5).

[0239] This study is a... 124An exploratory Phase 1 / 2 evaluation of I-p5+14 as an imaging agent for the safe and accurate detection of amyloid deposits in patients with systemic amyloidosis. The primary objective was to assess the safety and tolerability of the investigational reagent using systemic and organ-specific measures of dosimetry (Part 1) and standard measures of safety (vital signs, adverse events, and clinical laboratory measures).

[0240] Secondary objectives are extensively designed for evaluation:

[0241] 1. 124 Whether organ-specific retention of I-p5+14 can be observed in all patients with systemic amyloidosis in anatomical sites with biopsy-confirmed or clinically suspected amyloid deposits, and in subgroups of patients with each major amyloid type (e.g., AL, ATTR, ALECT2).

[0242] 2. In patients who can be appropriately evaluated, based on PET data, kidney and heart... 124 The correlation between I-p5+14 uptake (defined as the average and maximum concentration per unit volume [Bq / cc] or SUVR) and clinical indicators of organ function such as serum NTproBNP, creatinine, BUN, or eGFR.

[0243] 3. In subjects without abnormal amyloid deposition, i.e., in healthy controls, 124 Distribution of I-P5+14.

[0244] 4. In asymptomatic subjects with known transthyretin-amyloid mutations. 124 Biodistribution of I-p5+14.

[0245] 5. Repeat imaging can be used to understand whether the amyloid load in patients changes.

[0246] This study included n=50 patients with diagnosed and identified types of systemic amyloidosis, 2 asymptomatic subjects with confirmed TTR mutations at risk of amyloidosis (who were not evaluable and excluded from analysis based on organ- and patient-based sensitivity measures), and 5 healthy controls (a total of 57 unique subjects). Additionally, one participant from Part 2 successfully underwent repeated exposure. 124 I-p5+14 and subsequent imaging. Participants were not randomized or unaware of their disease status. Each participant underwent... 124I-p5+14 PET imaging agent. During the evaluation of PET / CT image data, the research nuclear medicine physician (reader) was unaware of the patient's or subject's diagnosis and suspected organ involvement.

[0247] This study comprised five parts. In Part 1 (n=3 patients with extensive AL amyloidosis), patients participated in a radiation dose escalation study to evaluate radiation dosimetry and pharmacokinetics. Patients received a single IV bolus. 124 I-p5+14 (<2 mg of p5+14 peptide). Patient 1 was given 0.31 mCi (0.84 mg p5+14), patient 2 was given 1.05 mCi (1.06 mg p5+14), and patient 3 was given 1.03 mCi (0.95 mg p5+14). Systemic and organ-specific dosimetry were estimated based on imaging data using the OLIDA-EXM method (1).

[0248] In Parts 2, 3, and 4, an additional 54 participants were included and received a single IV infusion. 124 I-p5+14 (1.98±0.19 mCi) 124 I and 1.42±0.19 mg p5+14). In Part 5, a single patient who was imaged during Part 2 received a second injection. 124 I-p5+14 (2.08 mCi and 1.49 mg p5+14) and repeated PET / CT scans.

[0249] Aseptic techniques were used in a biosafety cabinet within a certified cleanroom at the radiochemistry facility of the University of Tennessee Medical Center, Knoxville (research site) to synthesize radiolabeled pharmaceutical products. After testing for radiopurity, stability, bioactivity, and endotoxin levels, [the products were then processed]. 124 I-p5+14 doses were prepared in ≤35 ml of sterile phosphate-buffered saline containing 5% w / v human serum albumin (USP) as a stabilizer and 0.5 mg / ml ascorbic acid (USP) as an antioxidant. Participants received 30 ml of the solution delivered at a flow rate of 3 ml / min. 124 I-p5+14 was administered via IV infusion, followed by a 30 ml saline flush at the same rate. Iodine-124 was purchased from 3D Imaging (Little Rock, AR) and produced according to the information provided in DMF#025853. Table 4: Distribution of Subjects aOne patient was deemed ineligible due to the use of heparin, and four healthy volunteers were found to be ineligible due to comorbidities. Patient identification and imaging protocols

[0250] This study requires preliminary screening of medical records after obtaining a signed medical release to confirm the diagnosis of amyloidosis and assess clinical evidence of organ involvement in 13 predetermined organs and tissues. Figure 1 Following written consent, on day 1, participants underwent a physical examination and venipuncture, followed by a 7-day course of oral potassium iodide. Subjects received acetaminophen and antihistamines, administered via 30 ml IV infusion at 3 ml / min 30 minutes later. 124 I-p5+14, followed by a rinse with 30 ml of saline at the same rate.

[0251] On day 3, a second venous transection was performed. Patients with systemic amyloidosis and presymptomatic subjects were followed up by telephone on days 9 and 28. Healthy subjects enrolled after imaging of 42 patients were followed up for safety on days 3, 9, 28, and 56, and provided blood samples on days 28 and 56. Figure 1 ). Subject group

[0252] Participants were recruited locally and from around the United States. Patients included men and women >18 years of age diagnosed with a known type of amyloidosis. Healthy participants were >50 years of age, with no sex or race restrictions, no known germline mutations in the TTR gene, no diagnosis of amyloidosis or type 2 diabetes, and no confirmed or suspected first- or second-degree relatives with familial amyloidosis. One patient with AL and positive evidence of organ uptake underwent repeat imaging at ~24 months.

[0253] Patients who had been exposed to heparin within 7 days prior to infusion were excluded due to the known interaction between peptide p5+14 and heparin.

[0254] A total of n=25 patients with AL amyloidosis (n=23 with systemic AL and n=2 with localized AL); n=20 patients with ATTR amyloidosis (n=15 with ATTRV and n=5 with ATTRVT); two (n=2) patients with ALECT2 amyloidosis; and three (n=3) patients with other amyloid types were included for imaging (Table 3). Additionally, n=5 healthy volunteers and n=2 asymptomatic TTRV carriers were also included. Table 5: Demographic characteristics of participants at baseline Radioactive tracer synthesis and PET / CT imaging

[0255] Peptide p5+14 was synthesized by AmbioPharm Inc. (North Augusta, SC) and provided as a lyophilized powder in 3 mg aliquots, stored at -20°C. Iodine-124 was purchased from 3D Imaging (Little Rock, AR; DMF#025853). Patient-batch radiotracers were prepared on the day of use at the UT Radiochemistry Facility using a soluble iodogen as an oxidant. Images were acquired using a Siemens Biograph PET / CT (Siemens, Knoxville, TN) with ~30-second reduced-dose CT (120 kVp, 50 effective mAs). PET images were initially acquired from knee to ankle using 2 × 10-minute bed positions, followed by 5-minute bed positions from crown to thigh. Finally, 10-minute gated cardiac PET images were acquired.

[0256] PET data were reconstructed using a 3D OSEM algorithm with attenuation and instantaneous gamma correction, a 168×168 image matrix, and an image resolution of ~8 mm full width at half maximum. CT data were reconstructed using a medium smoothing kernel and a 4 mm reconstruction increment. Image Explanation

[0257] 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 prepared using Inveon Research Workplace (IRW) software (version 4.2 [4.2.0.15], Siemens Preclinical Solutions). Evaluation of the PET / CT images was performed by readers unaware of the clinical data. Radioactivity in organs was manually quantified by measuring radioactivity per unit volume (Bq / cc). 124 I-p5+14 uptake. The standard uptake ratio (SUVR) was determined for all organs by proportionally treating the mean radioactive concentration in a blood pool (radioactive chamber in the thoracic aorta).

[0258] Assess cardiac uptake in patients with AL- or ATTR-associated amyloidosis. 124I-p5+14 (SUVR) 平均 The correlation between renal radiotracer uptake and serum NT-proBNP during the same period was investigated. Similar correlations were found between renal radiotracer uptake and serum blood urea nitrogen (BUN), creatinine, and eGFR. end

[0259] The primary endpoint was determined based on the collection of adverse events (AEs) reported according to the Medical Dictionary for Regulatory Activities (MedDRA) after initial classification using CTCAE guidelines. Additional efficacy-based secondary and exploratory endpoints, as well as two measures of interest, were also investigated. Evaluate

[0260] Aside from serum biomarker measurements, no clinical assessment was performed as part of this study. During infusion... 124 Safety assessments were performed before and after I-p5+14. Vital signs were collected immediately before the infusion of the radiotracer and thereafter periodically every 50 minutes. Blood was drawn for clinical chemistry, complete blood count, C-reactive protein, lactate dehydrogenase, and NTproBNP tests. Figure 1 In healthy subjects, additional blood samples were drawn at 28 and 56 days post-infusion to assess anti-p5+14 antibodies using an internal peptide-capture ELISA. estimation of amyloid organ positivity

[0261] The factual criteria for diagnosis and organ involvement in this study were derived from medical records and based on standard diagnostic methods. Tissue biopsies were performed in most patients. Organ involvement was listed during pre-screening. Cardiac amyloidosis was typically diagnosed based on a combination of serum biomarkers (NT-proBNP), echocardiography, magnetic resonance imaging, nuclear imaging, and endocardial myocardial biopsy. The involvement of amyloid in other organs depends on a combination of organ-specific biopsy or remote biopsy with serum biomarkers. statistics

[0262] 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 participants, comprehensive subgroup analysis of patients with AL and ATTR, as well as healthy volunteers, was not feasible. All participants who completed the study were included in the image analysis, except for presymptomatic participants whose image quality was considered unacceptable.

[0263] By comparing PET / CT imaging in abdominal and thoracic organs 124Visual uptake of I-p5+14 and clinically known or anticipated organ involvement were used to determine patient- and organ-based sensitivity. Sensitivity and specificity (percentage 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). Similarly, patient-based sensitivity (i.e., positive amyloidosis in any organ or tissue) was calculated. 124 (Confirmed participants with I-p5+14 intake). Cardiac specificity was assessed in a healthy subject cohort where amyloid was assumed to be absent based on recruitment criteria.

[0264] Cardiac angiography was performed on patients and healthy subjects using an unpaired two-tailed t-test. 124 Comparison of I-p5+14 uptake, α=0.05. Spearman analysis (α =0.05) was used to determine renal or cardiac uptake. 124 The correlation between I-p5+14 uptake (SUVR mean) and BUN (mg / dL), creatinine (mg / dL), eGFR, and serum NT-proBNP (pg / mL) was calculated using SPSS Version 28 (Armonk, NY: IBM Corp.) and Prism. Moral supervision

[0265] This study was funded by the U.S. Food and Drug Administration and supported by an Investigational New Drug (IND) application (IND#132282). Further approvals were obtained from the Institutional Review Board (IRB) of the UT Graduate School of Medicine (Knoxville, TN). All patients provided written consent regarding their pre-screening medical records and imaging protocols prior to participation in the study. result Subject arrangement

[0266] Fifty-seven (n=57) participants were included (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 within 48 hours post-infusion. Subsequent participants (n=54) were administered 73.3 ± 7.0 mBq and 1.42 ± 0.19 mg of peptide, and PET / CT images were obtained 5.2 hours post-infusion (IQ, 4.93–5.42). The initial n=22 patients were also imaged within 24 hours post-infusion but were discontinued to minimize CT radiation dose, as the data did not contribute to the interpretation of findings at 5 hours post-infusion. All participants completed the study. Patient demographics and baseline characteristics

[0267] Fifty-one percent (51%) of the participants were male. The mean age of all participants was 64.8 ± 10.2 years. In the AL amyloidosis cohort, 52% (n=13 / 25) were of the λ isotype, and 40% (n=10 / 25) were of the k isotype. Eight percent (n=2 / 25) were not reported (Table 5). Patients with ATTR amyloidosis included 25% of ATTRwt (n=5 / 20). The ATTRV cohort included eight different germline mutations, the majority of which were T60A (n=6 / 20) (Table 5). Clinically, cardiac involvement was noted in 52% (n=13 / 25) of patients with AL or ATTR amyloidosis and 55% (n=11 / 20) of patients with AL or ATTR amyloidosis, respectively. Cardiac amyloidosis was clinically expected only in one of the other types of amyloidosis (ALYs). In the AL patient cohort, extracardiac amyloid was noted in the spleen (n=1 / 25), liver (n=3 / 25), and kidney (n=7 / 25) in medical records. Radioactive tracer production

[0268] Each batch was evaluated for its reactivity with amyloid fibrils. 124 I-p5+14, in which the average percentage of bound peptides is 97.5 ± 1.5%. In the prepared dosage... 124 The mean radioactivity purity of I-p5+14 is 98.8 ± 0.7%. All other disclosure criteria, particularly pyrogenicity, sterility, color, pH, peptide identity, peptide purity, and particle size, are in compliance with disclosure criteria. Dosage determination and pharmacokinetics

[0269] The estimated mean whole-body effective radiation dose for the sex was 0.23 (±0.02) mSv / MBq, with the highest organ dose received by the bladder being (0.94 ± 0.25 mSv / MBq). Radiation elimination primarily occurred via the renal and gastrointestinal pathways, with rapid loss of radiation from the central ventricle. The estimated whole-blood elimination half-life was 21.9 ± 7.6 hours. healthy subjects 124 Cardiac uptake and thorax-abdomen biodistribution of I-p5+14

[0270] In healthy subjects (n=5), radioactivity was observed in the stomach and bladder, as well as in the parotid and salivary glands, renal pelvis, ureter, and esophagus. In healthy subject number 4, radioactivity in the liver and kidneys was considered higher than the blood pool background, and therefore positive in these sites was considered. Figure 2A In the heart, no visible evidence of radiation was found in the LV or right ventricular (RV) wall or IVS in any healthy subject. Figure 2B ).therefore, 124 The visual evaluation of the I-p5+14 distribution yielded 100% specificity (negative percentage consistency) for the heart and spleen, and 80% specificity for the liver and kidney (Table 5). Table 6: Four major organs of healthy subjects 124 I-p5+14 intake among patients 124 Cardiac uptake of I-p5+14

[0271] PET / CT imaging was used to examine patients with ATTR or A1 amyloidosis. Figure 3A It can be readily seen in patients with AApoA1 and Alys-associated amyloidosis. 124 Cardiac uptake of I-p5+14. Cardiac uptake of the radiotracer was observed in patients with known cardiac involvement and in patients with unidentified cardiac involvement. Figure 3B In most cases, radioactivity is observed in the LV wall and IVS, with the RV wall and atria potentially affected.

[0272] In patients with AL and ATTR amyloidosis, the heart... 124The sensitivities for I-p5+14 uptake were 93% (n=13 / 14, [95% CI: 68.5–99.6]) and 100% (n=11 / 11, [95% CI: 74.2–100]), respectively. For the entire patient population, the cardiac sensitivity was 96% (n=25 / 26, [95% CI: 81.1–99.8]) (Table 6). This was observed in 27% (n=3 / 11) of AL patients and 67% (n=6 / 9) of ATTR patients. 124 Cardiac retention of I-p5+14, based on a review of medical records, indicated no clinical evidence of cardiac involvement in these patients. No gated cardiac imaging was evaluated. Table 7: Clinical evaluation and... for 13 evaluated organs and tissues in subjects with systemic amyloidosis 124 Consistency of amyloid distribution between I-p5+14 imaging assessments heart 124 Quantification of I-p5+14 uptake and its correlation with serum NTproBNP

[0273] Cardiac SUVR in AL and ATTR patient groups 平均 There is no difference (in the middle) Figure 4A In patients with systemic amyloidosis, cardiac uptake of the radiotracer was significantly higher than in healthy subjects. Figure 4A In the PET-positive (PET+) group, cardiac SUVR 平均 Significantly higher than PET-negative (PET-) patients ( Figure 4A ).

[0274] In patients with AL amyloidosis, cardiac SUVR was observed. 平均 A statistically significant correlation was found between NTproBNP and serum NTproBNP (rS=0.48, p=0.018). Figure 4B No correlation was observed in patients with ATTR amyloidosis (rS=0.28, p=0.231). Figure 4C In a post-hoc analysis of data from patients with AL amyloidosis, cardiac SUVR was proportionally processed using serum creatinine for each patient. 平均 This was used to explain variable renal function impairment. The composite variable was highly significantly correlated with serum NTproBNP (rS=0.61, p=0.0017). Figure 4D ). In the kidney SUVR 平均No correlation was observed with serum creatinine, BUN, or eGFR. among patients 124 Extracardiac biodistribution of I-p5+14

[0275] In 94% (n=44 / 47, [95% CI: 82.8–97.8]) of the enrolled patients, visual observation was observed in one or more of the predetermined organs in n=13. 124 I-p5+14 binding (Table 6). Two patients diagnosed with AL (n=2) and one patient with ATTR did not have [a specific finding] in any evaluated organ or tissue. 124 A positive I-p5+14 result is visually detectable. It has been 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). 124 I-p5+14 extracardiac conjugate ( Figures 5A-5B Despite a limited number of clinical observations, the sensitivity for liver and spleen amyloid was 100% (n=4 / 4, 95% CI: 51.0–100) and 100% (n=2 / 2, 95% CI: 17.8–100) in all patients (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). In patients without clinical evidence of organ-related involvement, amyloidosis was observed in the spleen and liver in 37.5% (n=9 / 24) and 18% (n=4 / 22) of patients with AL, respectively. 124 I-p5+14 uptake. Furthermore, in the absence of clinical evidence of organ involvement, it 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. 124 Intake of I-p5+14. Immunogenicity of p5+14

[0276] When compared with two control samples of normal human serum, there was no evidence of anti-peptide antibodies in either sample in the ELISA. Security

[0277] Safety analysis was performed on all subjects. Overall, 124I-p5+14 was well tolerated, with no deaths, drug-related serious adverse events (SAEs), or treatment-emergent adverse events (TEAEs) leading to study discontinuation (Table 8). Only one patient (1.8%, n=1 / 57) experienced a study drug-related TEAE, pruritus, and dysuria, both of which resolved without sequelae. Approximately half (49.1%, n=28 / 57) of the subjects reported non-study drug-related TEAEs, most of which were of mild severity. Table 8: Overall Overview of Adverse Events discuss

[0278] Once a diagnosis of amyloidosis is established, current anatomical imaging methods, including echocardiography and cardiac magnetic resonance imaging, provide valuable structural and functional information. 99m Tc-PYP and similar bone-finding agents are used to detect cardiac amyloid; however, they appear to bind to microcalcifications associated with amyloid deposits (Dietemann S, Nkoulou R). Ann Nucl Med. (2019;33:624–628), and their relationship with amyloid fibrils remains unclear. Therefore, a non-invasive imaging approach using reagents that directly bind to amyloid deposits is quantifiable and can detect cardiac amyloidosis before obvious anatomical changes and organ dysfunction, which would have significant clinical benefits.

[0279] Peptide p5+14 is a pattern recognition peptide for amyloid synthesis that binds to amyloid protein via multivalent electrostatic interactions (Antoni G, Lubberink M, Estrada S, ...). et al. J Nucl Med. 2013;54:213–220). In preclinical studies, the peptide was bound to synthetic amyloid-like fibrils and various types of amyloid-loaded tissue sections. When radiolabeled with iodine-124, the peptide is suitable for PET / CT imaging, providing quantitative, high-resolution biodistribution data. Iodine-124 is a non-residual, cyclotron-generated radionuclide with a half-life of 4.2 days, which can be enzymatically stripped from the peptide during renal catabolism by intracellular enzymes, causing physiological uptake in the gastric lumen, saliva, and thyroid gland (Cuddy SAM, Bravo PE, Falk RH, et al.JACC Cardiovasc Imaging. 2020;13:1325–1336). This procedure was used to allow for amyloid imaging in the kidneys (a clearance organ).

[0280] This refers to amyloidosis with eight different types (subtypes) – ALk, ALλ, ATTRwt, ATTRv, ALYs, AGel, ALECT2, and AApoA1. 124 I-p5+14 patients were included in the first human phase 1 / 2 study. The sensitivity of organ-specific uptake of the radiotracer was assessed by comparing data with those from the patients' medical records, which were used as the fact standard in this early study. Incidental imaging findings were also recorded following visual examination of PET images. Overall, a 2 mCi dose... 124 I-p5+14 (~1.4 mg peptide) was well tolerated with no deaths or drug-related SAEs. In subsequent studies, the 1 mCi... 124 The systemic effective dose (8.9 mSV) of I-p5+14 injection is equivalent to many thallium-201 and technetium-99m cardiac evaluations (Genovesi D, Vergaro G, Giorgetti A, et al. JACC Cardiovasc Imaging (2021;14:246–255). All patients diagnosed with amyloidosis in this study had confirmed [a specific abnormality] in at least one thoracic or abdominal organ or tissue (including the heart). 124 Visual uptake of I-p5+14 was observed in one AL patient diagnosed with cutaneous amyloid nodules and one renal amyloidosis patient who responded well to therapy and was presumed to have completely resolved amyloid deposits (18, 19). One patient diagnosed with systemic ATTRWT amyloidosis, peripheral neuropathy, and Congo red-positive ligamentum flavum was negative.

[0281] Cardiac uptake of the radiotracer was observed in ATTR patients with 100% clinical evidence supporting a diagnosis of cardiac amyloidosis. Furthermore, positive cardiac PET / CT imaging was observed in 6 ATTR patients without clinical evidence of cardiac involvement. This includes those with negative cardiac amyloidosis. 99m Patients with Tc-PyP imaging and normal serum NT-proBNP levels. These data suggest that... 124 PET / CT imaging with I-p5+14 can detect early, pre-symptomatic cardiac amyloid.

[0282] In the major thoracic and abdominal organs, clinical findings and 124The sensitivity (positive percentage concordance) between 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 simultaneously confirmed but relied on clinical reports in medical records, a limitation of the study design, but it indicates that cardiac amyloid persists even in patients who respond to treatment. Incidental uptake was observed in extracardiac locations via PET / CT imaging. 124 I-p5+14, particularly in the liver and spleen (organs that typically contain asymptomatic amyloid deposits) of patients with AL amyloidosis (Martin EB, Williams A, Richey T, et al.Sci Rep. 2016;6:22695). Although primarily considered a cardiac or neurological condition, ATTR amyloidosis is a systemic pathology. In patients with ATTR amyloidosis... 124 PET / CT imaging at I-p5+14 showed diverse anatomical uptake involving areas such as the lungs, liver, kidneys, pancreas, ball-and-socket joints, spine, and abdominal fat. These data illustrate... 124 I-p5+14 imaging can detect not only cardiac amyloidosis, but also extracardiac diseases that may lead to poor quality of life and affect the rate of cardiac amyloid removal in patients receiving amyloid degradation therapy.

[0283] PET imaging is an inherently quantitative modality. 124 I-p5+14 uptake can be readily assessed using various analytical methods, including determining organ-specific SUVR (a semi-quantitative measure in PET). In this study, this ratio was used to control for differences in radiotracer clearance, which can be significantly influenced by renal impairment, particularly in patients with AL amyloidosis and in many less common subtypes where renal involvement is prevalent. In this study, mean cardiac SUVR measurements were indistinguishable between the AL and ATTR populations. However, a small group of healthy controls showed significantly higher mean cardiac SUVRs in the ATTR and other amyloid populations.

[0284] Serum NT-proBNP is an empirically validated biomarker for overall survival in patients with AL amyloidosis. In these patients... 124 Cardiac retention at I-p5+14 was positively and significantly correlated with concurrent measurements, but this was not the case in patients with ATTR. 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).

[0285] Current amyloid therapies show a greater impact in patients with early-stage cardiac amyloidosis, thus necessitating early diagnosis. Agents that can directly detect amyloid deposits before symptom onset or when symptoms are mild may be advantageous. With the development of new amyloid clearance therapies, tools that can directly measure amyloid load, quantify changes, and demonstrate efficacy will be invaluable. A non-invasive and quantitative technique for detecting and monitoring cardiac and extracardiac deposits, such as using… 124 PET / CT imaging with I-p5+14 will be particularly useful. in conclusion

[0286] Data from this first human study supports 124 I-p5+14 has been used to detect the overall safety and efficacy of cardiac amyloidosis and systemic amyloid deposits. Based on these data, the United States and Europe have granted orphan designation for its use in diagnosing AL and ATTR amyloidosis. Example 2: Inclusion and Exclusion Criteria

[0287] This embodiment describes the inclusion and exclusion criteria for the studies disclosed in Embodiment 1.

[0288] The following inclusion and exclusion criteria were used in this study.

[0289] 1. Based on histological confirmation of apple green birefringence and Congo red deposition in the biopsy tissue, or the presence of amyloid-related pathology through genetic screening, or amyloid-specific imaging studies, the patient is diagnosed with systemic amyloidosis (SA). Additionally, the type of amyloidosis (AL, ATTR, ALECT2, or others) is characterized.

[0290] 2. Patients included in Part 1 (n=3) had extensive AL amyloidosis, defined as biopsy-confirmed or clinically detectable involvement of at least two organs (excluding the peripheral nervous system).

[0291] 3. All patients in Part 1 and Part 2 are 18 years of age or older. There are no gender or race restrictions.

[0292] 4. Women of fertility potential (who are not surgically sterilized, are not postmenopausal [generally understood as having last menstrual period > 2 years ago without drug intervention], and are of childbearing age) must test negative for pregnancy in a laboratory test performed by an on-site physician.

[0293] 5. Patients who are currently receiving or have received therapy or other drug-based anti-amyloid regimens may be included in the study (Part 1 and Part 2).

[0294] 6. The patient provides a signed, written, informed consent form, and meets the eligibility requirements, scheduled visits, and follow-up studies.

[0295] 7. Due to annual dose measurement restrictions, patients participating in another nuclear medicine amyloid imaging clinical trial protocol may be included in this study no earlier than 12 months after their previous radiotracer injection.

[0296] Prior to participating in Part 3 (n=2) of the trial, the following inclusion criteria were required:

[0297] 1. The patient has a well-defined germline mutation in the transthyretin (TTR) gene, putting them at risk of amyloidosis, and there is no clinical evidence of SA.

[0298] 2. All patients in Part 3 are >50 years old. There are no gender or race restrictions.

[0299] 3. Women of fertility potential (who are not surgically sterilized, are not postmenopausal [generally understood as having last menstrual period > 2 years ago without drug intervention], and are of childbearing age) must test negative for pregnancy in a laboratory test performed by an on-site physician.

[0300] 4. Subjects provide signed, written, and informed consent, and the study meets eligibility requirements, scheduled visits, and follow-up.

[0301] 5. Due to annual dose measurement restrictions, participants in another nuclear medicine amyloid imaging clinical trial protocol may be included in this study no earlier than 12 months after their previous radiotracer injection.

[0302] Prior to participating in Part 4 (n=5) of the trial, the following inclusion criteria must be met:

[0303] 1. Healthy control subjects (HC) are typically healthy adults, male or female, who have not been diagnosed with amyloidosis, do not have a first or second-degree relative (parent, sibling, child, aunt, uncle, niece, nephew) with confirmed or suspected familial amyloidosis, and do not have diabetes (e.g., type 2).

[0304] 2. All participants in Part 4 were 30 years of age or older.

[0305] 3. Women of fertility potential (who are not surgically sterilized, are not postmenopausal [generally understood as having last menstrual period > 2 years ago without drug intervention], and are of childbearing age) must test negative for pregnancy in a laboratory test performed by an on-site physician.

[0306] 4. The patient provides a signed, written, informed consent form, and meets the eligibility requirements, scheduled visits, and follow-up studies.

[0307] 5. Due to annual dose measurement limitations, patients participating in another nuclear medicine amyloid imaging clinical trial protocol may be included in this study no earlier than 12 months after their previous radiotracer injection.

[0308] Prior to participating in Part 5 of the trial (n=1), the following inclusion criteria are required:

[0309] 1. The patient has successfully completed Part 2 or Part 3 of the trial, and abnormal amyloid deposits in the thoracic and abdominal organs have been confirmed by CT / PET images.

[0310] 2. During Part 2 or Part 3 of the trial, the patient received a 2 mCi dose. 124 I-p5+14 provides visual evidence of radiotracer uptake in abdominal and thoracic organs associated with amyloid.

[0311] 3. Repeat exposure to the study reagent must occur at least 6 months after the first exposure.

[0312] 4. Patients submit serum samples for exploratory evaluation of the presence of p5+14 peptide-reactive antibodies, and the results are reviewed by the principal investigator before a second exposure to the study reagent.

[0313] 5. Patients meet all inclusion criteria for participation in Part 2 or Part 3, including signing informed consent for Part 5.

[0314] The following criteria excluded candidates from participating in Part 1 and Part 2 of the trial:

[0315] 1. Those with significant comorbidities (e.g., Eastern Cooperative Oncology Group [ECOG] score of 3 or higher), uncontrolled infections, or other serious illnesses.

[0316] 2. Patients with a persistent SpO2 ≤ 92% (as shown in medical records).

[0317] 3. Patients who require kidney dialysis.

[0318] 4. Women of reproductive potential (who are not surgically sterilized, are not postmenopausal [generally understood as having last menstrual period > 2 years ago without drug intervention], and are fertile) who test positive for pregnancy, are pregnant, or are breastfeeding in a laboratory test performed by an on-site physician.

[0319] 5. Patients who have received any amyloidophilic radiotracers as part of a research clinical trial (non-standard of care) within the past 12 months.

[0320] 6. Patients who have been exposed to heparin or heparin-based drugs within 7 days prior to the imaging study.

[0321] 7. Patients with known allergies to acetaminophen or iOSAT iodine therapy.

[0322] The following criteria exclude candidates from participating in Part 3 of the trial:

[0323] 1. Those with significant comorbidities (e.g., Eastern Cooperative Oncology Group [ECOG] score of 3 or higher), uncontrolled infections, or other serious illnesses.

[0324] 2. Subjects with a sustained SpO2 ≤ 92% (as shown in medical records).

[0325] 3. Women of reproductive potential (who are not surgically sterilized, are not postmenopausal [generally understood as having last menstrual period > 2 years ago without drug intervention], and are fertile) who test positive for pregnancy, are pregnant, or are breastfeeding in a laboratory test performed by an on-site physician.

[0326] 4. Subjects with clinical evidence of amyloidosis based on standard clinical guidelines.

[0327] 5. Subjects suffering from polyneuropathy of unknown origin.

[0328] 6. Subjects who have received any amyloid-positive radiotracers as part of a study clinical trial (non-standard of care) within the past 12 months.

[0329] 7. Subjects who were exposed to heparin or heparin-based drugs within 7 days prior to the imaging study.

[0330] 8. Subjects with known allergies to acetaminophen or iOSAT iodine treatment.

[0331] 9. Subjects with clinical signs of SA based on routine surveys of serum and urine biomarkers, radiographic or nuclear imaging studies, or peripheral nerve evaluations.

[0332] The following criteria excluded participants from Part 4:

[0333] 1. Those with significant comorbidities (e.g., an Eastern Cooperative Oncology Group [ECOG] score of 2 (limited physical activity but not bedridden, and able to perform light or sedentary work, such as light housework or office work) or higher), uncontrolled infections, or other serious illnesses.

[0334] 2. Individuals with a sustained SpO2 ≤ 92% (as shown in medical records).

[0335] 3. Women of reproductive potential (who are not surgically sterilized, are not postmenopausal [generally understood as having last menstrual period > 2 years ago without drug intervention], and are fertile) who test positive for pregnancy, are pregnant, or are breastfeeding in a laboratory test performed by an on-site physician.

[0336] 4. Subjects who have received any amyloid-positive radiotracers as part of a study clinical trial (non-standard of care) within the past 12 months.

[0337] 5. Subjects who were exposed to heparin or heparin-based drugs within 30 days prior to the imaging study.

[0338] 6. Subjects with known allergies to acetaminophen or iOSAT iodine treatment.

[0339] 7. Subjects who have been diagnosed with type 2 diabetes or are taking medication to manage type 2 diabetes.

[0340] 8. Active infections that cannot be completed before day 1 of the study and require systemic antiviral or antimicrobial therapy.

[0341] 9. A known history of human immunodeficiency virus (HIV), hepatitis C, or chronic hepatitis, or a positive test result.

[0342] 10. Uncontrolled hypertension (BP>160 / 100 mm Hg).

[0343] 11. Smoking more than 20 cigarettes a day.

[0344] 12. Diagnosed as heart failure with preserved ejection fraction.

[0345] 13. Any other medical condition that the investigator believes will make the subject unsuitable for inclusion or may interfere with the subject's participation in or completion of the study.

[0346] The following criteria excluded participants from Part 5:

[0347] 1. Any patient who has experienced a clinically significant adverse event related to prior exposure to the study drug.

[0348] 2. Any subject who meets any exclusion criteria (if applicable) in Part 2 or Part 3. Example 3: Using 124 I-P5+14 ( 124 I-p5+14) versus 18F-flubetapyr cardiac amyloid quantification: Preliminary PET / CT study

[0349] This example describes the quantification of myocardial function in participants and controls with amyloid cardiomyopathy (CMP). 124 I-p5+14 intake and its comparison with 18 A preliminary study of the diagnostic value of F-flubetapyr, the main objective of which was (1) to quantify in participants and controls with amyloid CMP. 124 I-p5+14 cardiac uptake, (2) comparison 124 I-p5+14 and 18 The diagnostic value of F-flubetapyr in Al-CMP and AttrWT-CMP, and (3) measurement 124 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. method Participation

[0350] The study was approved by the Mass General Brigham Human Research Committee, and each participant provided written informed consent (Protocol No. 2021P000085). 124 The 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, 46 participants were included: 12 with AL-CMP, 12 with ATTRwt-CMP, 2 with hereditary amyloid CMP (1 with the Thr60Ala [P.T80A] variant ATTR [ATTRv], and 1 with apolipoprotein A-IV amyloidosis [AApoAIV]), and 12 control participants without amyloidosis. Additionally, participants from previous treatments were included. 18 Eight control participants in the F-flubetapyr PET / CT study (Dorbala S, Vangala D, Semer J, et al.Eur J Nucl Med Mol Imaging.2014;41:1652–1662). One AL-CMP participant due to... 124 Acquisition errors during I-p5+14 imaging resulted in exclusion. Participants with hereditary amyloidosis were excluded from 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 biopsy confirming amyloid type by immunohistochemistry or mass spectrometry, and cardiac involvement demonstrated by imaging or endocardial myocardial biopsy. Grade 2 / 3 cardiac involvement was assessed. 99m Tc-pyrophosphate ( 99m Tc-PYP SPECT / CT scans are used to diagnose ATRwt amyloidosis, and serum free light chain assays and serum and urine immunofixation electrophoresis are used to rule out AL amyloidosis, or by endocardial biopsy using immunohistochemistry or mass spectrometry. Hereditary amyloid CMP is diagnosed by biopsy using immunohistochemistry or mass spectrometry; if appropriate, variant TTR is identified by gene sequencing, and cardiac involvement is confirmed by imaging or endocardial biopsy. PET / CT acquisition

[0351] use 124 I-p5+14 and 18 F-flubetapyr PET / CT for molecular imaging of amyloidosis in participants with amyloid CMP (except for one Al-CMP participant who can only undergo this procedure). 124 I-p5+14 PET / CT). A control group underwent one PET / CT scan: 12 patients used... 124 I-p5+14, 8 users 18 F-Fluorbetapi. Imaging was performed using a Discovery MI PET / CT scanner (GE Healthcare, Chicago, IL, USA), with CT scout used for patient localization and low-dose chest CT scans used for attenuation correction and image fusion. Acquisition and quantification methods were specific for each radiotracer, taking into account the differences in radiotracer characteristics. Intravenous injection was administered over 5 minutes. 124I-p5+14. Based on previous data on radiotracer kinetics, cardiac PET was performed only 30 minutes in static mode 5 hours post-injection (to allow for clearance of blood pool activity). The median net injection activity, limited by concerns about exposure to participants and staff, was 1.00 mCi (interquartile range [IQR] 0.94–1.07), with a median effective dose of 9.01 mSv (IQR 8.55–9.47, based on 0.23 mSv / MBq, including 0.5 mSv for low-dose CT scans). We used 130 mg potassium iodide to protect the thyroid for 7 days, starting 1 day prior to radiotracer injection. 18 F-fluorobetapi was administered as a bolus injection 1 minute after PET acquisition began in list mode. Static images of the heart were reconstructed using data from 4 to 30 minutes post-injection of the radiotracer, based on previous publications. The median net active dose 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 for low-dose CT scans). 18 Following F-flubetapyr scan, the median interval was 1.5 days (IQR 1–4.5, range 1–12 days). 124 I-p5+14 scan. Quantitative PET / CT

[0352] Myocardial uptake was measured volumetrically on static images using PMOD software (PMOD Technologies LLC, Zürich, Switzerland). Because manually tracing the myocardial contour on each slice lacked reproducibility and was inaccurate in controls, we used automated iso-contouring in PMOD to exclude ventricular blood pools and reproducibly tracing the myocardium. First, the target volume (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 cavity blood pool, and the interventricular septum was attributed to the LV tracing. Specific iso-contouring thresholds were selected for each radiotracer due to differences in radiotracer kinetics and target-to-background ratios. Blood pool activity concentrations were measured in a 10 mm diameter spherical VOI in the left atrium (LA). 18 For F-flubetapyr, we used a threshold of 2 × mean blood pool active concentration, as in our previous study. 124 I-p5+14, we used the mean of the blood pool active concentration + 2 standard deviations as a threshold, which better depicts myocardial volume, especially in mild cases. Figure 6 ).

[0353] In order to 124 For automatic isocontour mapping of I-Evuzamitide PET images, we first manually traced the target volume (VOI) on the fused PET / CT images to define the overall left and right ventricular (LV and RV) contours. Then, we measured the blood pool active concentration in a 10 mm diameter spherical VOI in the left atrium (LA). Next, we used the isocontour mapping function of PMOD software with a threshold of the mean ± 2 standard deviations (SD) of the LA blood pool active concentration, which appropriately depicts myocardial volume under various uptake levels. Conversely, in the last column, with a threshold of 2 × the mean LA blood pool active concentration, as used for 18E-flubenzurid, the relevant portion of the myocardial volume is ignored. This is our approach. 124 The reason for using a specific threshold for I-evuzamitide (LA+2SD).

[0354] Our primary LV and RV uptake measures are percentage injected dose (%ID), calculated as mean VOI active concentration × VOI volume / injected activity. This measure is adjusted for injected activity but not for body weight, which is unnecessary for radiotracers accumulating in the heart and specific organs rather than throughout the body. We also analyzed standardized uptake (SUV). 平均 SUV 最大 Cardiac amyloid activity (CAA) is calculated as average or maximum VOI activity concentration / (injected activity / body weight) VOI SUV 平均 ×VOI volume, and target to background ratio (TBR) as mean VOI active concentration / mean blood pool active concentration (same as the SUV ratio, where corrections for injected activity and body weight in the SUV are offset in this ratio). In recipient operating characteristic (ROC) analysis, a threshold for abnormally high uptake values ​​is defined by maximizing the Youden index (sensitivity + specificity - 1) to identify the cutoff with optimal sensitivity and specificity, thereby classifying CMP cases and controls, and assigning equal weights to false positives and false negatives. Structural and functional markers of amyloid load

[0355] A search was conducted within one year of PET / CT scans of clinically performed echocardiography (N=23), cardiac MRI (N=13), and... 99m Reports and images of Tc-PYP SPECT (n=11) (if available). For Level 3, include older reports. 99mTc-PYP SPECT results were as expected, with no further changes anticipated. The median time difference for PET / CT was 1.7 months (IQR 0.7–4.1) for echocardiography, 2.7 months (IQR 0.7–5.0) for MRI, and 4.5 months (IQR 2.5–13.8) for SPECT. For interventricular septal thickness, LV mass index, ejection fraction, myocardial contractile fraction (MCF = stroke volume / myocardial volume), global longitudinal strain (GLS), late gadolinium enhancement, ECV, and... 99m The Tc-PYP classification was used for evaluation. Laboratory tests, functional status and quality of life

[0356] 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 class (NYHA) and the 6-minute walk test. Quality of life was measured using the Minnesota Heart Failure Life Questionnaire (MLWHFQ, 21 questions, total score 0-105, higher scores indicate worse symptoms), the Kansas City Cardiomyopathy Questionnaire (KCCQ, 23 questions, total score 0-100, higher scores indicate fewer symptoms), and the 36-Item Short Form Survey (SF-36, 36 questions, converted T-scores used for normalized population 0-100, higher scores indicate better quality of life). Statistical analysis

[0357] Continuous variables were represented as medians with IQR and compared using the Kruskal-Wallis test, followed by the Dunn test for pairwise comparisons. We included adjustments for multiple tests using the Benjamini-Hochberg procedure. Categorical variables were represented as frequencies as percentages and evaluated using Fisher's exact test. Paired variables were compared as needed using the Wilcoxon signed-rank test or the McNemar test. Correlation was quantified using Spearman's ρ with Fieller correction at 95% CI. We present two-tailed p-values, and they are considered statistically significant if < 0.05. 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), using the packages tidyVerse, Desctools, Gtsummary, Rstatix, Proc, and Correlation.d. result Patient characteristics

[0358] In the amyloid CMP participants (N=24), the median age was 74 years (IQR 69–78) and 22 were male (92%). The ATTRwt-CMP participants (N=12) were older than the AL-CMP participants (N=12), had lower heart rates, and worse cardiac parameters such as LV quality index, ejection fraction, and GLS. However, other demographic, clinical characteristics, quality of life, functional status, and biomarkers were similar (Table 9). At study inclusion, 10 AL-CMP participants were in hematologic remission following plasma cell-directed therapy (83%), 2 were receiving therapy but not in remission (17%), and 10 ATTRwt-CMP participants received tafamidix (83%). In the controls (N=20), the median age was 62 years (IQR 58–67) and 13 were male (65%). Table 9: Participant Characteristics 124 I-p5+14 PET / CT LV and RV myocardial uptake

[0359] The radioactive tracer injection was well tolerated in all participants, and no adverse reactions were noted. Amyloid CMP was observed in all participants regardless of amyloid type. 124 Significant cardiac uptake was observed in I-p5+14 participants, while no visual cardiac uptake was observed in any of the control participants. Furthermore, in equivocal grade 1... 99m Among the participants in Tc-PYP SPECT / CT, 124 I-p5+14 PET / CT confirmed clear myocardial uptake, which was confirmed by endocardial myocardial biopsy showing TTR amyloid deposits. Figure 7 ). 124 I-p5+14 LV %ID differed between groups (overall p<0.001): AL-CMP median 1.48 (IQR 1.12–1.89), ATTRwt-CMP 2.12 (1.66–2.47), and control 0.00 (0.00–0.01, Table 8). Several uptake parameters tended to be higher in ATTRwt-CMP than in AL-CMP, particularly LVSUV. 平均 and RV index ( Figure 8 (Table 10). Based on the Youden index, high LV %ID, LV CAA, and LV TBR completely distinguished amyloid CMP cases from controls, while LV SUV 平均 SUV 最大 The RV index shows some overlap ( Figure 8 (Table 10). In fact, some control participants exhibited heterogeneous background activity, with small spots of activity concentration exceeding the automatic isopleth mapping threshold based on the blood pool. This resulted in SUVs within the amyloid CMP range. 平均 However, this does not visually indicate amyloid deposition. In contrast, %ID and CAA were close to zero in control participants because these metrics include VOI volume, thus amyloid deposits in CMP cases were better distinguished from heterogeneous background activity in control participants. Table 10: 124 I-p5+14 and 18 F-flubetapyr myocardial uptake index 18 F-flubetapyr PET / CT LV and RV myocardial uptake

[0360] In the same amyloid CMP participants (except one who did not use it) 18In AL-CMP participants undergoing F-flubetapyr PET / CT, and in different control participants, we found that... 18 F-Fluorbetapyr and its use 124 The results for I-p5+14 are similar ( Figure 9 (Table 10). 18 F-flubetaprazole LV %ID also differed between groups (overall p<0.001): AL-CMP median 1.03 (IQR 0.78–2.03), ATTRwt-CMP 1.10 (0.89–1.44), and controls 0.04 (0.00–0.29). LV myocardial uptake indices did not differ between AL-CMP and ATTRwt-CMP participants, but the range was higher for AL-CMP. High LV %ID and LVCAA completely distinguished amyloid CMP cases from controls, while LV SUV... 平均 The LVTBR and RV indices show some overlap. use 124 I-p5+14 comparison 18 F-flubetapyr myocardial radiotracer uptake index

[0361] When comparing LV myocardial radiotracer uptake among the same participants, in AL-CMP, the following was used: 124 I-p5+14 and 18F-flubetapyr for LV and RV %ID, SUV 平均 SUV 最大 It is similar to CAA, but in ATTRwt-CMP, it adopts... 124 I-p5+14 is higher (all p≤0.002, Figure 10 (Table 10). In AL-CMP and ATTRwt-CMP, the LV and RV TBRs using F-fluorobetapyr were compared with those using... 124 The higher values ​​of I-p5+14 (all p≤0.002) reflect the different blood and tissue kinetics of the two tracers.

[0362] 124 Correlation between I-p5+14 intake and direct and indirect markers of amyloid load, functional status, and quality of life

[0363] Throughout the group, we found 124I-p5+14 LV %ID showed a moderate correlation with NYHA grade (ρ=0.47), MLWHFQ total score (ρ=0.59), KCCQ total score (ρ=-0.62), SF-36 physical T score (ρ=-0.50), and SF-36 mental T score (ρ=-0.41), and a strong correlation 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 The correlation between F-flubetapyr LV %ID (ρ=0.50), interventricular septal thickness (echocardiography ρ=0.78, MRI ρ=0.58), LV mass index (echocardiography ρ=0.70, MRI ρ=0.82), LV MCF (echocardiography ρ=-0.66, MRI ρ=-0.62), LV GLS (ρ=0.54), and LVECV (ρ=0.51) was moderate to strong. Table 11: 124 Correlation between I-P5+14 intake and direct and indirect markers of amyloid load, functional status, and quality of life discuss

[0364] This preliminary study quantified a novel ubiquitin-binding radiotracer. 124 I-p5+14 myocardial uptake in participants with amyloid CMP, its diagnostic value and 18 F-flubetapyr was compared, and its concentration in amyloid CMP was measured. 124 The correlation between I-p5+14 myocardial uptake and markers of amyloid burden was observed. In all participants with AL, ATTRwt, ATTRv, or AApoAIV CMP, the correlation was... 124 Visual interpretation of I-p5+14 PET / CT images identified cardiac amyloidosis. This was observed in all participants with AL-CMP or ATTRwt-CMP. 124 The quantitative myocardial uptake in both I-p5+14 LV and RV was significantly higher than in control participants. Importantly, 124 I-p5+14 LV %ID, CAA, and TBR completely distinguished all cases and controls with known AL-CMP and ATTRwt-CMP. Similar results were observed with 18L-flubetapyr in participants with the same amyloid CMP, except for TBR, supporting this finding. 124The effectiveness of I-p5+14 detection. Therefore, LV %ID and CAA are the most accurate indicators for identifying amyloid CMP using two radiotracers. When comparing radiotracers, besides TBR, 124 I-p5+14 consistently showed higher myocardial values ​​in ATTRwt-CMP. Furthermore, in both A1-CMP and ATTRwt-CMP participants, 124 I-p5+14LV %ID showed a moderate to strong correlation with indirect indicators of amyloid burden from echocardiography and MRI (interventricular septal thickness, LV mass index, MCF, GLS, ECV), indicating effective quantification of amyloid burden. Furthermore, 124 I-p5+14LV %ID and 18 F-Fluorobetapine LV %ID is moderately correlated, indicating differences in binding sites, binding affinities, and / or quantification of different components of amyloid deposits. In fact, 124 I-p5+14 is specifically designed for binding negatively charged surfaces of glycosaminoglycans and amyloid fibrils, 19 and 18 F-Fluorobetaphene is a stilbene with a structure similar to thioflavin T, known to bind to the surface of β-sheets along channels formed by cross-strand ladders within amyloid protoplasts. 27 124 The correlation between I-p5+14 LV %ID and ECV was also moderate, likely because ECV measures fibrosis, inflammation, and / or edema in addition to amyloid deposits. The correlation between I-p5+14 LV %ID and ECV was moderate, and the correlation with cardiac biomarkers was strong, further supporting the adoption of... 124 The effectiveness of I-p5+14 PET / CT in quantifying cardiac amyloid. Finally, 124 I-p5+14 injections were well tolerated in all participants, and no side effects were noted.

[0365] 124 I-p5+14 is a novel radiotracer based on the synthetic peptide p5+14, developed to bind to the negatively charged surfaces of glycosaminoglycans and amyloid fibrils in all types of amyloid deposits. This peptide has been successfully tested in a variety of radiotracers in a mouse model of serum amyloid A amyloidosis and in humans with multiple types of systemic amyloidosis. This study systematically evaluated [the results] in a cohort of participants with amyloid CMPs derived from multiple precursor proteins. 124 The ability of I-p5+14 to be taken up by the myocardium is novel, and it is the first time that it has been compared with... 18Comparison with F-flubetapyr. Our findings support the use of... 124 I-p5+14 PET / CT imaging accurately and safely diagnoses AL-CMP and ATTRwt-CMP, and aids in the identification of hereditary amyloid CMP. These results suggest that this panamyloid-binding radiotracer has the potential to reduce the need for endocardial myocardial biopsy to diagnose cardiac amyloidosis, limiting biopsies to patients with positive scans who require confirmation of amyloid type. Our results also indicate that... 124 I-p5+14 can quantify cardiac amyloid burden. In fact, LV... 124 I-p5+14 uptake is a structural and functional measure of LV (lower ventricle) and amyloid load, and is associated with cardiac biomarkers and quality of life related to heart failure. Pre-treatment use... 124 Imaging with I-p5+14 can identify and quantify the presence of amyloid in the heart and other organs, particularly for predicting potential major effects and organ-specific side effects, while post-therapy imaging can measure organ response. Although ECV may be an alternative method for quantifying amyloid load in the heart, it is not specific to amyloidosis and can also represent inflammation or fibrosis, the latter being a process triggered by macrophage-mediated amyloid removal. Therefore, targeted imaging of cardiac amyloid using specific molecular radiotracers can offer advantages over other methods.

[0366] Recently, PET radiotracers targeting β-amyloid protein ( 11 C-Pittsburgh-B-compounds 18 F-Fluorbetapyr 18 F-Fluorbetaben and 18 F-fluorometholone has emerged as a useful tool for evaluating Al-CMP. However, the ability of these radiotracers to detect ATTR-CMP is limited, especially... 18 F-flubetapyr and 18 F-Fluorobetaben. Non-invasive diagnosis of ATTR-CMP is typically based on SPECT bone affinity (bone-avid) radiotracers (F-Fluorobetabenzene). 99m Tc-pyrophosphate[ 99m Tc-PYP] 99m Tc-3-Diphospho-1,2-propanedicarboxylic acid [ 99m Tc-DPD] 99m Tc-hydroxymethylene diphosphonate [ 99m[Tc-HMDP]. These radiotracers have reportedly exhibited near 100% specificity for grade 2 / 3 uptake, but only 74% sensitivity for ATTR-CMP. In fact, bone affinity SPECT radiotracers may lack sensitivity in detecting early-stage disease in ATTR-CMP and are unreliable for diagnosing Al-CMP, certain forms of ATTRv-CMP, and other rare forms of amyloid CMP. Furthermore, these data are derived from high-volume centers assessing selected patients with advanced disease or a high pre-test probability, potentially overestimating the diagnostic performance of SPECT in non-specialized centers. Additionally, continuous quantitative testing with bone affinity radiotracers is limited by the relatively low sensitivity of SPECT compared to PET and the early stages of developing quantitative SPECT methods. Importantly, the myocardial uptake mechanism of bone affinity radiotracers remains unclear, impairing the interpretation of post-treatment changes. In contrast, PET pan-amyloid-binding radiotracers... 124 I-p5+14 provides higher quantitative accuracy than SPECT bone affinity radiotracers and is more sensitive for ATTR-CMP than β-amyloid-specific PET radiotracers. Notably, in this study, we found that in ATTRwt-CMP participants, using 124 I-p5+14 significantly increased myocardial uptake compared to those using [other methods]. 18 F-Fluorobetapine. These participants also tended to show higher levels of F-fluorobetapine than AL participants. 124 I-p5+14 uptake, and using 18 F-flubetapirid showed more similar uptake across amyloid types. Meanwhile, in terms of echocardiography and cardiac MRI, ATTRwt-CMP participants had worse structural and functional parameters, such as LV mass index and GLPS, than AL-CMP participants. In ATTRwt-CMP... 124 Higher myocardial uptake of I-p5+14 can better reflect the severity of cardiomyopathy and can be compared with... 18 F-Fluorbetapyr provides more sensitive diagnosis of early disease. There is a great need for techniques that provide specific and quantitative estimates of both cardiac AI and ATTR amyloid load to evaluate organ responses to amyloid-depleting therapies. 124 I-p5+14 can fill this gap.

[0367] In short, 124I-p5+14 PET / CT imaging is a promising new radiotracer capable of accurately detecting various types of cardiac amyloid deposits, distinguishing between the presence and absence of cardiac amyloidosis, and quantifying cardiac amyloid burden. In ATTRwt-CMP, it is used... 124 Quantitative amyloid protein levels using I-p5+14 can be compared to those using... 18 F-flubetapyr is more accurate. Larger, multicenter studies are needed to validate these findings and further evaluate the results. 124 Clinical utility of I-p5+14 PET / CT imaging for early diagnosis and treatment monitoring of cardiac amyloidosis. Example 4: By 124 PET / CT imaging at I-p5+14 to monitor the progression and regression of amyloid.

[0368] This embodiment describes the situation when suffering from different types of diseases. Systemic amyloidosis In patients, a novel amyloid-reactive peptide called P5+14, labeled with iodine-124, was detected. 124 Semi-quantitative analysis of PET / CT imaging of I-p5+14 was used to monitor the progression and regression of amyloid. method

[0369] This is a multipart, open-label study to evaluate 124 Safety, dosing assays, and biodistribution of the I-p5+14 peptide in patients with any type of systemic amyloidosis; presymptomatic subjects carrying germline TTR mutations; and healthy subjects as described in Example 1. In this example, 9 patients with AL and 10 patients with ATTR underwent repeat PET / CT imaging for... 124 The organ-specific uptake of I-p5+14 peptide relative to baseline was visualized and quantified. The imaging protocol is described in Example 1.

[0370] Quantification of organ radioactivity by measuring the standard uptake ratio (SUVR) and the amount of radioactivity per unit volume proportionally treated by blood pool radioactivity (Bq / cc) 124 Uptake of I-p5+14 peptide. The correlation between SUVR and serum NTproBNP, left ventricular (LV) wall and interventricular septum (IVS) thickness, and overall longitudinal strain was calculated. result

[0371] In nine patients with AL amyloidosis, organ-specific uptake ratios (SUVR) and serum NT-proBNP levels were measured. 124 Changes in I-p5+14 peptide uptake showed that... Figure 11A In the AL patient population, 1 / 9 (11%) showed SUVR.平均 The SUVR decreased to near normal levels, with 7 / 9 (78%) showing stable cardiac function. 平均 1 / 9 (11%) showed increased cardiac SUVR 平均 NTproBNP remained unchanged, and 8 / 9 (89%) showed stable NTproBNP, of which 1 / 9 (11%) showed increased NTproBNP.

[0372] In 9 patients with ATTR amyloidosis, organ-specific uptake ratios (SUVR) and serum NTproBNP were measured. 124 Changes in I-p5+14 peptide uptake showed that... Figure 11B In the ATTR patient population, 2 out of 10 (20%) showed cardiac SUVR. 平均 A reduction of more than 20%, 7 / 10 (70%) showed stable SUVR 平均 1 / 10 (10%) showed increased cardiac SUVR 平均 NTproBNP remained unchanged, and 9 / 10 (90%) showed stable NTproBNP levels, of which 1 / 10 (10%) showed a decrease in NTproBNP.

[0373] In all patients, the changes in NTproBNP between imaging time points and 124 Changes in cardiac uptake of I-p5+14 are related (Table 12, Figure 12 When each group was assessed separately, no changes in serum NTproBNP were detected in either AL or ATTR patients. 124 Correlation between changes in cardiac uptake of I-p5+14. Table 12: Changes in serum NT-proBNP and 124 Correlation between changes in cardiac uptake of I-p5+14

[0374] In AL patients, an association was detected between SUVR and LV wall and GLS, but not with IVS. Figure 13A -C). In ATTR patients, an association was detected between SUVR and LV wall and IVS, but not with GLS ( Figure 13D -F). For all patients, a correlation was detected between SUVR and LV wall, IVS, and GLS (-F). Figure 13G -I). discuss

[0375] This example demonstrates the use of PET / CT imaging. 124Semi-quantitative analysis of I-p5+14 uptake allows for monitoring the progression and regression of amyloid, as well as a way to study the relationship between amyloid, amyloidosis, organ function, and patient outcomes. Example 5: Early Cardiac Amyloid Detection via PET / CT Imaging with 125I-p5+14

[0376] This study is 124 A phase 3 evaluation of the efficacy of I-p5+14 in diagnosing cardiac amyloidosis. The primary objective was to evaluate the efficacy of I-p5+14 based on visual scan interpretation. 124 I-p5+14 PET / CT imaging is used to diagnose cardiac amyloidosis with high sensitivity and specificity.

[0377] A secondary objective was to evaluate the incidence of treatment-induced adverse events (AEs) and changes in clinical laboratory values ​​from baseline by tracking the changes from day 1 to day 30. 124 Safety of a single intravenous administration of I-p5+14.

[0378] The purpose of exploration is: 1) evaluation 124 The efficacy of I-p5+14 in diagnosing cardiac ATTR amyloidosis; 2) Evaluation 124 The efficacy of I-p5+14 in diagnosing cardiac AL amyloidosis, 3) Evaluation 124 The efficacy of I-p5+14 in differentiating cardiac amyloidosis from AL and ATTR; 4) Evaluation 124 Quantitative cardiac uptake of I-p5+14 correlated with amyloid content in endocardial myocardial biopsies, cardiac biomarkers, clinical stage, participant-reported heart failure health status, and the use of standardized measures of structure and function in imaging; 5) evaluate participants' responses to biopsy or multimodal imaging and 124 Preferred I-p5+14 PET / CT imaging. Inclusion and Exclusion Criteria

[0379] The following inclusion criteria were used in this study.

[0380] 1) Understand the research procedures and be able to provide a signed informed consent form (as specified in Section 10.1.3), which includes compliance with the requirements and limitations listed in the Informed Consent Form (ICF) and this protocol.

[0381] 2) Male or female, ≥18 years old.

[0382] 3) Suspected cases of cardiac amyloidosis who are willing to undergo diagnostic evaluation for cardiac amyloidosis (e.g., echocardiography, CMR, bone affinity tracer cardiac SPECT, extracardiac or endocardial myocardial biopsy, etc.). Participants can be enrolled before or during their diagnostic evaluation for cardiac amyloidosis.

[0383] 4) Ability to perform PET / CT imaging as part of a study, including the ability to lie supine for approximately 1 hour.

[0384] 5) For women of childbearing potential: consent is required during the treatment period and administration of medication. 124 After I-p5+14, abstain from sexual intercourse for at least 30 days or use a contraceptive method with a failure rate of <1% per year. a. A woman is considered to have reproductive potential if she has reached menopause (≥12 months of amenorrhea without a definite cause other than menopause) and has not undergone surgical sterilization (removal of the ovaries and / or uterus). The definition of reproductive potential may be adjusted according to local guidelines or requirements. b. Examples of contraceptive methods with a failure rate of <1% per year include bilateral tubal ligation, male sterilization, established and appropriate use of ovulation-suppressing hormonal contraceptives, hormone-releasing intrauterine devices (IUDs), and copper IUDs. i. Contraceptive methods that do not produce a failure rate of <1% per year, such as caps, diaphragms, or sponges with spermicide, or male or female condoms with or without spermicide, are unacceptable. ii. The reliability of abstinence should be evaluated based on the duration of the clinical trial and the participants' preferred and usual lifestyles. Regular abstinence (e.g., calendar method, ovulation method, symptom-basal body method, or post-ovulation method) and withdrawal are unacceptable methods of contraception.

[0385] 6) For men: consent to abstinence or use of contraception and consent to avoid donating sperm, as defined below: a. For female partners of fertility potential, men must abstain from sex or use condoms plus additional contraception, which together result in a failure rate of <1% per year during the treatment period and for at least 120 days (30 days plus a 90-day spermatogenesis cycle) after the last dose of the study intervention. Men must also avoid donating sperm during the same period.

[0386] The following exclusion criteria were used in this study.

[0387] 1) Confirm the diagnosis of cardiac amyloidosis.

[0388] 2) Confirm a diagnosis of systemic amyloidosis without known cardiac involvement (e.g., renal AL or ATTR peripheral neuropathy). Participants with only carpal tunnel tissue or laminectomy who are amyloid-positive are permitted, provided they do not have other known organ involvement.

[0389] 3) At enrollment, patients were receiving approved treatments (e.g., tafamadis) or were in a clinical trial for the treatment of ATTR cardiac amyloidosis. For AL and ATTR: patients may enter the screening period for clinical trials after signing an ICF and participants are enrolled in the trial, but may not receive the experimental therapy until after the safety follow-up on day 30 of the trial.

[0390] 4) Pregnancy or breastfeeding.

[0391] 5) Incapacitated mentally or legally, exhibiting significant emotional problems at the time of the study, or having a history of mental illness.

[0392] 6) Has a known allergy to potassium iodide.

[0393] 7) Undergo hemodialysis or peritoneal dialysis.

[0394] 8) eGFR less than 15 ml / min / 1.73m2.

[0395] 9) Myocardial infarction within three (3) months of screening.

[0396] 10) Suffering from severe claustrophobia or any medical condition that would prevent the imaging protocol from being completed.

[0397] 11) Having any disease that the researchers believe may confound the study results or pose additional risks to the participants.

[0398] 12) In 124 Heparin or heparin analogues (e.g., enoxaparin, dalteparin, fondaparin) have been administered within seven (7) days prior to I-p5+14 administration.

[0399] 13) Known uncorrected thyroid conditions (except for mild elevation of thyroid-stimulating hormone with normal T4). Subject population and identification

[0400] This study will include participants suspected of having cardiac amyloidosis. To ensure that the inclusion requirements for the primary endpoint are met, inclusion 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 included. An independent clinical adjudication committee with expertise in cardiac amyloidosis will be established to review information obtained during diagnostic evaluation (e.g., clinical history and laboratory findings, endocardial biopsy, extracardiac biopsy, echocardiography, cardiac MR, and bone affinity tracer cardiac SPECT) to determine whether the data support a diagnosis of cardiac amyloidosis.

[0401] The screening period lasts up to 30 days. Eligibility criteria will be confirmed and informed consent will be obtained. For all WOCBPs administered, the treatment will be given to women of reproductive potential (WOCBP). 124 A serum pregnancy test (immediate serum β-hCG pregnancy test) must be obtained before I-p5+14. In all WOCBPs, the pregnancy test result must be available and... 124 The test result was negative before administration of I-p5+14.

[0402] The critical period for treatment is from day 1 to day 30. If in I... 124 If any nuclear medicine study (e.g., SPECT using a bone affinity tracer) is performed prior to I-p5+14 administration, then the Day 1 visit should not be earlier than 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 study, then wait until day 5 or later for a follow-up visit to allow for further observation. 124 Minimizing I-p5+14 scattering into SPECT scans may be necessary.

[0403] The SoT (Sort of True Time) diagnostic evaluation period is used to diagnose cardiac amyloidosis. If all required diagnostic tests are not completed by day 30, the diagnostic evaluation period is extended until either the last diagnostic test is completed or until day 60, whichever is earlier. Research intervention and imaging protocols

[0404] During the first day of visit, the subject will receive a single dose of [treatment / treatment]. 124 I-p5+14, at a dose level of 1 mCi (±10%), wherein not more than 2 mg of p5+14 is administered via intravenous push at a rate of ~1 mL / 5 seconds. 124 I-p5+14.

[0405] In application 124 Before I-p5+14: 1) A serum pregnancy test will be obtained (immediate serum β-hCG pregnancy test), and after administration... 124 Prior to I-p5+14, all WOCBP tests were confirmed to be negative. 2) Participants will begin treatment on day 1 with oral KI 130 mg once daily (e.g., iOSAT, potassium iodide oral solution USP, 65 mg / ml) for 3 days. 124 3) Start more than 30 minutes before I-p5+14 to reduce radioactive iodine uptake in the thyroid gland; 4) Record heart rate, blood pressure, weight, height, and accompanying medications; 5) A cohort of approximately 25 participants will have triplicate 12-lead electrocardiograms (ECGs) used to assess the effects of I-p5+14 administration. 124The corrected QT (QTc) interval was obtained within 2 hours prior to I-p5+14.

[0406] exist 124 After administration of I-p5+14: 1) A cohort of approximately 25 participants with baseline QTc assessment will receive three copies of a 12-lead ECG; 2) Participants will undergo PET / CT scans.

[0407] PET / CT imaging was performed on day 1 following administration of 124I-p5+14. Participants will be administered... 124 Four hours (±60 minutes) after I-p5+14, a PET / CT scan was performed from the base of the crown to the upper thigh. The total PET / CT scan time was estimated at 45 minutes, including 20 minutes in the cardiac bed position.

[0408] All participants will have a baseline 12-lead ECG obtained during screening. A cohort of approximately 25 participants will have triplicate ECGs for assessment of administration. 124 Within 2 hours before I-p5+14 124 Immediately after I-p5+14 administration and administration 124 QTc interval 30 (+10) minutes after I-p5+14. Image Explanation

[0409] Based on (i) independent reader visual evaluation of PET / CT images and (ii) cardiac amyloid disease status based on clinically accurate criteria, participants were categorized into one of the following groups: a) True positive (TP): having positive results from PET / CT images. 124 Participants with visually positive cardiac uptake of I-p5+14 and confirmed as positive cardiac amyloidosis by factual criteria, b) True negative (TN): those with images from PET / CT. 124 Participants with visually negative cardiac uptake (I-p5+14) and confirmed as having negative cardiac amyloidosis by fact criteria, c) False positives (FP): those with images from PET / CT images 124 Participants with visually positive cardiac uptake (I-p5+14) and confirmed negative cardiac amyloidosis by fact criteria, d) False negatives (FN): those with negative results from PET / CT images. 124 Participants with visually negative cardiac uptake of I-p5+14 and confirmed positive cardiac amyloidosis by factual criteria.

[0410] Sensitivity is defined as the number of TP participants divided by the sum of TP and FN participants: Sensitivity = TP / (TP + FN).

[0411] Specificity is defined as the number of TN participants divided by the sum of TN and FP participants: Specificity = TN / (TN + FP). end

[0412] The main purpose of this study is to evaluate 124 The efficacy of I-p5+14 in diagnosing cardiac amyloidosis. The corresponding co-primary endpoints are as follows: 1) ... 124 PET / CT imaging at I-p5+14 has improved sensitivity for diagnosing cardiac amyloidosis based on visual scan interpretation. 2) Using 124 PET / CT imaging at I-p5+14 has specificity for diagnosing cardiac amyloidosis based on visual scan interpretation. The common principal assumptions are: 1) Invalid: using... 124 Imaging at I-p5+14 has a sensitivity of 0.65 for diagnosing cardiac amyloidosis. Alternatively, one could use... 124 Imaging at I-p5+14 showed a sensitivity >0.65 for diagnosing cardiac amyloidosis, and 2) was ineffective: using 124 Imaging at I-p5+14 has a specificity of 0.55 for diagnosing cardiac amyloidosis; alternatively: use 124 Imaging at I-p5+14 has a specificity >0.55 for diagnosing cardiac amyloidosis. Sensitivity and specificity will be calculated for each of the three readers. The study will be considered successful if the null hypothesis for both sensitivity and specificity is rejected by at least two of the three readers. Sensitivity and specificity will be summarized using the Exact method with two-sided 95% confidence intervals (CI). For each of sensitivity and specificity, the lower limit of the two-sided 95% CI will be mathematically higher than the threshold of 65% (sensitivity) or 55% (specificity) for the common primary endpoint if the above null hypothesis is rejected by an exact binomial test. Subgroup analyses of the primary endpoint will be performed using two-sided 95% confidence intervals (α=0.025) for the following groups: 1) type of systemic amyloidosis (AL, ATTR), 2) sex (female, male), 3) age (age >65, age ≤65). Example 6: Compared with 99mTc-PYP, by 124 Early cardiac amyloid detection on PET / CT imaging at I-p5+14

[0413] In order to evaluate the use 124 The potential consequences of switching between PET / CT imaging diagnostic methods of I-p5+14 and 99mTc-PYP, using 124PET / CT imaging at I-p5+14 re-evaluated 24 individuals previously tested for cardiac amyloidosis. The procedure was performed according to the method described in Example 1. 124 PET / CT imaging of I-p5+14.

[0414] The results are shown in Table 13 below. Of the 7 individuals who tested positive for cardiac amyloid using the 99mTc-PYP test, all 7 individuals used... 124 The I-p5+14 test was positive for cardiac amyloid. Of the 17 individuals who tested negative for cardiac amyloid using the 99mTc-PYP test, 11 used... 124 PET / CT imaging at I-p5+14 showed a positive result for cardiac amyloid. These results confirm that, compared to 99mTc-PYP, using... 124 I-p5+14 increases the sensitivity of cardiac amyloid detection. Table 13: Use 124 I-p5+14 and 99mTc-PYP detection of cardiac amyloid protein Example 7: Using 124 Diagnostic threshold for I-p5+14 uptake

[0415] Organ-specific SUVR was calculated using computer vision analysis of PET / CT images from each individual, in 16 individuals without systemic amyloidosis and 27 individuals with systemic amyloidosis (AL, ATTR, or others). 124 Organ-specific uptake of I-p5+14. Organ-specific SUVR values ​​were calculated for the liver, spleen, kidneys, pancreas, heart, and adrenal glands.

[0416] Automatic organ segmentation was used to quantify cardiac uptake of radiotracers. The heart and aorta were automatically segmented on CT data using a 3D pre-trained convolutional neural network. Contours were applied to PET data to quantify mean normalized uptake values ​​and other uptake parameters in the heart.

[0417] By calculating organ-specific SUVR for healthy individuals 平均 The 97.5th percentile of the value is used to calculate the SUVR below each organ-specific value. 平均 The diagnostic threshold for the value was calculated. The same method was also used to calculate the SUVR value for each organ. 最大 Standard deviation of uptake in organs (SUVR) hetero ), SUVR 峰 The sum of all uptakes in an organ normalized by voxel size of the image (SUVR) 总 ) and SUVR heteroDivide by SUVR 平均 Calculated coefficient of variation (SUVR) cov The diagnostic threshold of ).

[0418] If amyloid protein is detected in an organ at a level higher than the threshold in Table 14, the individual is diagnosed with systemic amyloidosis. Table 14: SUVR Thresholds of Organs

Claims

1. A method for early diagnosis of amyloid-related disease in individuals at risk of developing the disease, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b. The amount of amyloid reactive peptides can be detected by measuring the amount of a detectable marker in an individual's tissues or organs. An amount of amyloid reactive peptides above the threshold indicates that an individual has amyloid-related disease, which is in the early stage of amyloid-related disease.

2. The method of claim 1, wherein an individual is identified as being 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 from carpal tunnel release surgery, having monoclonal gammopathy of undetermined significance (MGUS), having heart failure with preserved ejection fraction (HFpEF), having heart failure with reduced ejection fraction (HFrEF), being from a susceptible racial group, or being older.

3. The method of claim 2, wherein the gene mutation is in transthyretin.

4. The method of claim 2, wherein the gene mutation is in fibrinogen α protein.

5. The method of any one of claims 1-3, wherein the individual does not have symptoms of amyloidosis.

6. The method of any one of claims 1-3, wherein the individual has neuropathological symptoms of amyloidosis.

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

8. A method for early diagnosis of amyloid-related disease in individuals suspected of having it, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b. The amount of amyloid reactive peptides can be detected by measuring the amount of a detectable marker in an individual's tissues or organs. An amount of amyloid reactive peptides above the threshold indicates that an individual has amyloidosis, with amyloid-associated disease being in its early stages.

9. A method for determining the prognosis of an individual with amyloid-associated disease, comprising: a. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 for the first time; b. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to quantify amyloid in the individual a second time. and c. Compare the amyloid protein quantified for the first time in a tissue or organ with the amyloid protein quantified for the second time in a tissue or organ to determine the individual's prognosis.

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

11. The method of claim 8, wherein the prognosis of an individual suffering from amyloid-related disease is based on the detection of the amount of amyloid reactive peptides in the heart and kidneys.

12. The method of claim 8, wherein the prognosis of an individual suffering from amyloid-related disease is based on the detection of the amount of amyloid reactive peptides in the heart, kidneys, and all other organs.

13. A method for treating amyloid-related diseases, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence shown in any one of SEQ ID NO: 1-14 conjugated to a detectable marker; b. Detecting the amount of amyloid reactive peptides by measuring the amount of detectable markers in an individual's tissues or organs, and c. If the amount of amyloid reactive peptide is above the threshold, it is used for the treatment of amyloid-related diseases.

14. The method of claim 13, wherein the individual suffers from early amyloid-related disease.

15. A method for monitoring the treatment of amyloid-related diseases in an individual, comprising: a. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 for the first time; b. For the treatment of amyloid-related diseases; c. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to a second quantification of amyloid in the individual; and d. Compare the amyloid protein measured for the first time in a tissue or organ with the amyloid protein measured for the second time in the tissue or organ to determine whether the treatment is effective.

16. The method of any one of claims 1-15, further comprising administering an amyloid reactive peptide to an individual, the amyloid reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the amyloid reactive peptide by detecting the amount of the detectable marker in the individual's tissues or organs, to quantify amyloid in the individual a second, optionally a third, fourth, and / or fifth time.

17. The method of claim 15 or 16, wherein the first and second times are spaced at least six weeks apart.

18. A method for selecting individuals for the treatment of amyloid-related diseases, comprising: a. Administering an amyloid-reactive peptide to an individual, said amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14; and b. The amount of amyloid reactive peptides can be detected by measuring the amount of a detectable marker in an individual's tissues or organs. If amyloid protein is detected in the heart, treatment for amyloid-related diseases is administered; if amyloid protein is not detected in the heart, replacement therapy is administered.

19. A method for managing the treatment of amyloid-related diseases in an individual, comprising: a. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, 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 for the first time; b. For the treatment of amyloid-related diseases; c. Administering an amyloid-reactive peptide to an individual, the amyloid-reactive peptide comprising an amino acid sequence conjugated to any one of SEQ ID NO: 1-14, and detecting the amount of the detectable label in the individual's tissues or organs to quantify amyloid in the individual a second time; d. Compare the amyloid protein levels quantified for the first time in a tissue or organ with those quantified a second time in the same tissue or organ; and e. Adjust the treatment used for amyloid-related diseases.

20. The method of claim 19, wherein if the amyloidosis is stable between the first and second treatments, the treatment is changed.

21. The method of claim 19, wherein treatment is restarted if the amyloid load increases.

22. The method of claims 9-12 and 14-21, wherein an amount of amyloid reactive peptide above a threshold indicates that an individual has an amyloid-related disease.

23. The method of claim 22, wherein the amyloid-associated disease is in the early stage of amyloid-associated disease.

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

13.

25. The method of any one of claims 1-24, wherein the detectable marker is a radioactive marker.

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

27. The method of any one of claims 1-26, wherein the detectable marker is 124 I.

28. The method of any one of claims 1-27, wherein the dose administered to the individual is from about 0.1 to about 25 mCi. 124 I-labeled amyloid reactive peptide.

29. The method of any one of claims 1-28, wherein the dose administered to the individual is about 0.3 mCi, 1 mCi, or about 2 mCi. 124 I-labeled amyloid reactive peptide.

30. The method of any one of claims 1-29, wherein the dose administered to the individual is about 1 mCi (±10%). 124 I-labeled amyloid reactive peptide.

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

32. The method of any one of claims 1-31, wherein an individual is administered about 0.3 mg, 1 mg, 1.5 mg, or about 2 mg of an amyloid reactive peptide.

33. The method of any one of claims 1-32, wherein the amyloid reactive peptide is administered intravenously.

34. The method of any one of claims 1-33, further comprising determining the organ-specific standard uptake ratio for an individual's internal organs.

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

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

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

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

39. The method of any one of claims 1-38, wherein the sensitivity for 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%.

40. The method of any one of claims 1-39, wherein the sensitivity is about 99% or about 100%.

41. The method of any one of claims 1-40, 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 of any one of claims 1-41, wherein the specificity of the amyloid reactive peptide is about 99% or 100%.

43. The method of any one of claims 1-42, further comprising measuring one or more health-related quality of life measures for an individual.

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

45. The method of claim 44, wherein the biomarkers associated with amyloid-related diseases are selected from the group consisting of troponin T, NTproBNP, urinary protein levels, UACR, EGFR, and alkaline phosphatase levels.

46. ​​The method of any one of claims 1-45, wherein the tissue or organ of the individual is selected from the group consisting of lung, fat, heart, kidney, pancreas, joint, spine, liver, spleen, adrenal gland, bone lesion, choroid plexus, pituitary gland, uterus, bone marrow, musculoskeletal tissue, gastrointestinal tract and prostate.

47. The method of claims 1-8, 13 and 22-46, wherein the threshold is the average threshold of SUVR.

48. The method of claim 47, wherein if the organ or tissue is the liver, the average 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 average SUVR threshold is approximately 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5; and if the organ or tissue is the kidney, the average SUVR threshold is approximately 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, or 2.3; If the organ or tissue is the pancreas, the average 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 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 of claim 48, wherein the average SUVR threshold is 1.31 if the organ or tissue is the liver, 1.21 if the organ or tissue is the spleen, 1.88 if the organ or tissue is the kidney, 1.40 if the organ or tissue is the pancreas, 1.19 if the organ or tissue is the heart, or 1.35 if the organ or tissue is the adrenal gland.

50. The method of claims 1-8, 13 and 22-46, wherein the threshold is the maximum threshold of SUVR.

51. The method of claim 50, wherein 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, or 16. 0, 10.5, 11, 11.5 or 11.7; 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, the maximum SUVR threshold is 4.9, 5, 5.5, 6, 6.5, 7 or 7.5; or if the organ or tissue is the adrenal gland, the maximum SUVR threshold is 1, 1.1, 1.2, 1.3, 1.4, 1.5 or 1.

6.

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

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

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

55. The method of any one of claims 1-54, further comprising performing additional amyloid imaging of the individual prior to the application of the amyloid reactive peptide.

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

57. The method of claim 56, wherein further imaging of the individual's amyloid protein comprises a tracer selected from the group consisting of: 99m Tc-PyP, 99m Tc-DPD, 99m Tc-HMDP, 99m Other bone scintillation tracers for Tc-MDP 124 I, and 18 F-Fluorbetapyr 18 F-Fluorometabolone and 18 F-Fluorobetabenzene.

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

59. The method of any one of claims 1-58, wherein the individual is diagnosed with amyloid cardiomyopathy.

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

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

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

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

64. The method of any one of claims 1-63, wherein the amyloid reactive peptide binds to amyloid proteins including: 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), colloidin (AGel), apolipoprotein C-II (AApoCII), and apolipoprotein C. -II (AApoCIII), lysozyme (ALys), leukocyte chemotactic factor (ALECT2), fibrinogen a variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactoglobulin (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.

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

66. The method of any one of claims 1-65, wherein amyloid-associated diseases are selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AApoCII, AApoCIII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AIAPP, APro, AIns, APrP, and Aβ amyloidosis.

67. The method of claim 66, wherein the amyloid-associated disease is cardiac AL amyloidosis.

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

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

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

71. The method of any one of claims 1-70, wherein the individual is an elderly person.

72. The method of any one of claims 1-71, wherein the individual suffers from early amyloid-related disease.

73. The method of any one of claims 1-72, wherein the individual suffers from early systemic amyloidosis.

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

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

76. The method of claim 75, wherein the early AL amyloidosis is diagnosed according to the Mayo clinical system.

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

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

79. The method of claim 78, wherein early ATTR amyloidosis includes stage 1 ATTR amyloidosis.

80. The method of any one of claims 1-12, 16-18 and 20-79, further comprising administration for treatment of amyloid-related diseases.

81. The method of any one of claims 13-15, 19, and 80, wherein the treatment for amyloid-related diseases is selected from the group consisting of: transthyretin stabilizers (e.g., tafamidix, aclamidix), transthyretin silencers (e.g., Partisan, Inothesen, Vutrisan, Eploetersen), gene editing methods, anti-amyloid methods using monoclonal antibodies, treatments targeting plasma cell clones (e.g., daratumumab, bortezomib), and antibody-peptide fusions comprising an antibody-peptide fusion protein, said antibody-peptide fusion protein comprising a second amyloid reactive peptide and an antibody binding to amyloid fibrils.

82. The method of claim 81, wherein the second amyloid reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-14 containing 0, 1, 2, 3 or 4 amino acid substitutions, insertions or deletions.

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

84. The method of any one of claims 81-83, wherein the antibody-peptide fusion protein comprises: (i) an amyloid reactive peptide comprising the amino acid sequence shown in SEQ ID NO: 2; and (ii) An antibody that binds to human amyloid fibrils, 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 the VH comprises: CDR-H1 comprising the amino acid sequence shown in SEQ ID NO: 106, CDR-H2 comprising the amino acid sequence shown in SEQ ID NO: 107 and CDR-H3 comprising the amino acid sequence shown in SEQ ID NO: 108, and the VL comprises: CDR-L1 comprising the amino acid sequence shown in SEQ ID NO: 103, CDR-L2 comprising the amino acid sequence shown in SEQ ID NO: 104 and CDR-L3 comprising the amino acid sequence shown in SEQ ID NO: 105; wherein the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain, and wherein the amyloid reactive peptide is linked to the antibody via a spacer comprising the amino acid sequence shown in SEQ ID NO:

83.

85. The method of any one of claims 1-84, wherein an individual is given a peptide comprising an amyloid-reactive peptide and an N-terminal leader sequence.

86. The method of claim 85, wherein the peptide comprising the amyloid reactive peptide comprises the amino acid sequence shown in SEQ ID NO:

100.

87. The method of claim 86, wherein the detectable marker binds to one or more amino acids at positions 1 to 6 of the amino acid sequence shown in SEQ ID NO:

100.

88. The method of claim 87, wherein the detectable marker is 124 I.

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

100.

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

91. The method of claim 90, wherein administering potassium iodide comprises administering 130 mg of potassium iodide to the patient for seven days, starting one day prior to administering the amyloid reactive peptide.

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

93. The method of any one of claims 90-92, wherein the potassium iodide is administered orally.

94. A peptide comprising the amino acid shown in SEQ ID NO: 100, wherein the peptide comprises a detectable tag binding to one or more amino acids at positions 1 to 6.

95. The peptide of claim 94, wherein the detectable marker is 124 I.

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

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