Compositions and methods for detecting ccr2 receptors

Abstract

Among the various aspects of the present disclosure is the provision of compositions of imaging agents and methods for use in detecting, monitoring, and evaluating CCR2 associated diseases, disorders, and conditions.

What is claimed is: 
     
         1 . An imaging agent comprising:
 a CCR2 binding peptide;   a radiolabel; and   a nanoparticle, a chelator, or a linker.   
     
     
         2 . The imaging agent of  claim 1 , further comprising a linker. 
     
     
         3 . The imaging agent of  claim 1 , wherein the CCR2 binding peptide comprises:
 (i) a linear ECL1 peptide or a cyclized ECL1i peptide;   (ii) an amino acid sequence, Thr-Phe-Leu-Lys (SEQ ID NO: 17);   (iii) an amino acid sequence, SEQ ID NO: 17, comprising one or more chemical modifications that confers resistance to proteolysis;   (iv) an amino acid sequence, SEQ ID NO: 17, comprising one or more conservative substitutions;   (v) an amino acid sequence, Thr-Phe-Leu-Lys-Cys (SEQ ID NO: 1);   (vi) an amino acid sequence, SEQ ID NO: 1 comprising one or more chemical modifications that confer resistance to proteolysis;   (vii) an amino acid sequence, SEQ ID NO: 1 comprising one or more conservative substitutions;   (viii) an amino acid sequence, X1-TFLKC-X2 (SEQ ID NO: 2), wherein X1 is absent, is glycine, or represents an amino acid sequence selected from the group consisting of AG, LG, YLG, and HYLG; and X2 independently is absent, is methionine, or represents an amino acid sequence selected from the group consisting of MA, MAN, MANG, MANGF, MANGFV, MANGFVW, MANGFVWE, and MANGFVWEN;   (ix) an amino acid sequence, SEQ ID NO: 2 comprising one or more chemical modifications that confer resistance to proteolysis;   (x) an amino acid sequence, SEQ ID NO: 2 comprising one or more conservative substitutions;   (xi) an amino acid sequence, X1-TFLK-X3 (SEQ ID NO: 18), wherein X1 is absent, is glycine, or represents an amino acid sequence selected from the group consisting of AG, LG, YLG, and HYLG; and X3 independently is absent or is alanine;   (xii) an amino acid sequence, SEQ ID NO: 18, comprising one or more chemical modifications that confer resistance to proteolysis; and   (xiii) an amino acid sequence, SEQ ID NO: 18, comprising one or more conservative substitutions.   
     
     
         4 . The imaging agent of  claim 1 , wherein
 the CCR2 binding peptide comprises amino acids and all or a portion of the amino acids are in L configuration or in D configuration;   the imaging agent is stored in a physiological pH, optionally, at about pH of 7.4;   the CCR2 binding peptide is covalently linked to the nanoparticle; or   the CCR2 binding peptide is no more than 18 amino acids in length.   
     
     
         5 . The imaging agent of  claim 1 , wherein the CCR2 binding peptide is selected from the group consisting of:
 LGTFLKC (SEQ ID NO: 3);   HYLGTFLKCMA (SEQ ID NO: 4);   LGTFLKCMA (SEQ ID NO: 5);   HYLGTFLKC (SEQ ID NO: 6);   GTFLKCMANGF (SEQ ID NO: 7);   TFLKCMANGFV (SEQ ID NO: 8);   HYLGTFLKCMANGFVWEN (SEQ ID NO: 9);   LGTFLK (SEQ ID NO: 19);   AGTFLKC (SEQ ID NO: 20);   LGTFLKA (SEQ ID NO: 21);   GTFLK (SEQ ID NO: 22);   AGTFLKA (SEQ ID NO: 23);   a sequence deriving from any of SEQ ID NO: 3 to 10, or 19 to 23 by one or more chemical modifications that confer resistance to proteolysis; and   a sequence deriving from any of SEQ ID NO: 3 to 9 or 19 to 23 by one or more conservative substitutions.   
     
     
         6 . The imaging agent of  claim 5 , wherein the CCR2 binding peptide consists of LGTFLKC (SEQ ID NO: 3). 
     
     
         7 . The imaging agent of  claim 1 , wherein
 (i) the radiolabel comprises  2 H (D or deuterium),  3 H (T or tritium),  11 C,  13 C,  14 C,  64 Cu,  67 Cu,  177 Lu,  13 N,  15 N,  15 O,  17 O,  18 O,  18 F,  89 Sr,  35 S,  153 Sm,  36 Cl,  82 Br,  75 Br,  76 Br,  77 Br,  123 I,  124 I,  125 I,  131 I,  111 In,  67 Ga,  68 Ga,  177 Lu,  186 Re,  188 Re,  201 Tl  99m Tc,  90 Y, or  89 Zr;   (ii) the radiolabel comprises oxygen-15 water, nitrogen-13 ammonia, [ 82 Rb] rubidium-82 chloride, [ 11 C], [ 11 C] 25B-NBOMe, [ 18 F] Altanserin, [ 11 C] Carfentanil, [ 11 C] DASB, [ 11 C] DTBZ, [ 18 F]Fluoropropyl-DTBZ, [ 11 C] ME@HAPTHI, [ 18 F] Fallypride, [ 18 F] Florbetaben, [ 18 F] Flubatine, [ 18 F] Fluspidine, [ 18 F] Florbetapir, [ 18 F] or [ 11 C] Flumazenil, [ 18 F] Flutemetamol, [ 18 F] Fluorodopa, [ 18 F] Desmethoxyfallypride, [ 18 F] Mefway, [ 18 F] MPPF, [ 18 F] Nifene, Pittsburgh compound B, [ 11 C] Raclopride, [ 18 F] Setoperone, [ 18 F] or [ 11 C] N-Methylspiperone, [ 11 C] Verapamil, [ 11 C] Martinostat, Fludeoxyglucose ( 18 F)(FDG)-glucose analogue, [ 11 C] Acetate, [ 11 C] Methionine, [ 11 C] Choline, [ 18 F] Fluciclovine, [ 18 F] Fluorocholine, [ 18 F] FET, [ 18 F] FMISO, [ 18 F] 3′-fluoro-3′-deoxythymidine, [ 68 Ga] DOTA-pseudopeptides, [ 68 Ga] PSMA, or [ 18 F] Fluorodeoxysorbitol (FDS);   (iii) the chelator comprises NHS-MAG 3 , MAG 3 , DTPA, 3p-C-NE3TA, 3p-C-NOTA, 3p-C-DE4TA, ATSM, tetraazamacrocyclic ligands (e.g., DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTA-NHS, pSCN-Bn-DOTA, pNH 2 -Bn-DOTA, TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid, TETA-octreotide (OC)), hexaazamacrobicyclic cage-type ligands (e.g., Sarcophogine chelators), cross-bridged tetraamine ligands (e.g., CB-TE2A (4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane)), 6-Hydrazinopridine-3-carboxylic acid (Hynic), NHS-Hynic, 2,2′,2″-(10-(2-((2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (Maleimido-mono-amide-DOTA); or   (iv) the nanoparticle comprises a nanocluster or nanostructure; organic, inorganic, or lipid nanostructures;   (v) the nanoparticle comprises iron oxide, gold, gold nanoclusters (AuNC), gold nanorods (AuNR), copper (Cu), quantum dots, carbon nanotubes, carbon nanohorn, gadolinium (Gd), dendrimers, dendrons, polyelectrolyte complex (PEC) nanoparticles, calcium phosphate nanoparticles, perfluorocarbon nanoparticles (PFCNPs), lipid-based nanoparticles, liposomes, or micelles; or   (vi) the linker comprises a chemical or physical bond; PEG, TA-PEG-Maleimide, TA-PEG-OMe, TA-PEG, an isothiocyanate group, a carboxylic acid or carboxylate groups, a dendrimer, a dendron, Fmoc-protected-2,3-diaminopropanoic acid, ascorbic acid, a silane linker, minopropyltrimethoxysilane (APTMS), dopamine, 2 thiol groups, 2 primary amines, a carboxylic acid and primary amine, maleimide and thiol, hydrazide and aldehyde, or a primary amine and aldehyde, an amide, a thioether, a disulfide, an acetyl-hydrazone group, a polycyclic group, a click chemistry (CC) group.   
     
     
         8 . The imaging agent of  claim 1 , wherein
 (i) the chelator is conjugated to the CCR2 binding peptide and the chelator is radiolabeled; or   (ii) the CCR2 binding peptide is conjugated to a nanoparticle.   
     
     
         9 . The imaging agent of  claim 8 , wherein the radiolabel is  64 Cu. 
     
     
         10 . The imaging agent of  claim 8 , wherein the CCR2 binding peptide is conjugated to a nanoparticle comprising a gold nanocluster. 
     
     
         11 . The imaging agent of  claim 10 , wherein the gold nanocluster is loaded with a radiolabel. 
     
     
         12 . The imaging agent of  claim 1 , wherein the chelator comprises a tetraazamacrocyclic ligand; DOTA; TETA; or 2,2′,2″-(10-(2-((2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (Maleimido-mono-amide-DOTA). 
     
     
         13 . The imaging agent of  claim 1 , wherein the chelator is conjugated to a cysteine residue of the CCR2 binding peptide. 
     
     
         14 . The imaging agent of  claim 1 , wherein the imaging agent comprises:
 (i) a  64 Cu-DOTA-ECL1i PET or SPECT imaging agent;   (ii) an ECL1i peptide conjugated to a gold nanocluster, wherein the gold nanocluster is loaded with  64 Cu;   (iii)  64 Cu-DOTA-ECL1i;   (iv)  64 CuAuNCs-ECL1i;   (v) a multivalent imaging agent (optionally,  64 CuAuNCs-ECL1i), exhibiting extended pharmacokinetics for long-term CCR2 receptor detection and targeted theranostics; or   (vi) a monovalent (optionally,  64 Cu-DOTA-ECL1i), exhibiting fast pharmacokinetics for efficiently for rapid or serial imaging of CCR2 receptors.   
     
     
         15 . The imaging agent of  claim 1 , wherein the imaging agent
 is a PET imaging agent;   is a SPECT imaging agent;   targets CCR2 receptors;   detects CCR2 receptor up-regulation; or   detects elevated CCR2 expression.   
     
     
         16 . A method of detecting a CCR2 receptor comprising:
 administering to a subject an imaging agent of  claim 1 ; and   detecting the imaging agent.   
     
     
         17 . The method of  claim 16 , wherein detecting the CCR2 receptor comprises positron emission tomography (PET) imaging, and single photon emission computed tomography (SPECT) imaging, mass spectrometry, gamma imaging, magnetic resonance imaging (MRI), magnetic resonance spectroscopy, fluorescence spectroscopy, CT,
 ultrasound, or X-ray.   
     
     
         18 . The method of  claim 17 , wherein the method detects a CCR2 associated disease, disorder, or condition is selected from the group consisting of:
 (i) atherosclerosis; abdominal aortic aneurysm; acquired metabolic disease; acute cystitis; acute lung injury; acute proliferative glomerulonephritis; acute or chronic sinusitis; age-related macular degeneration; alcoholic hepatitis; allergic asthma; allergic conjunctivitis; allergic rhinitis; alveolitis; angiostenosis; anthracosis; ariboflavinosis; arteriosclerosis; artery disease; arthritis; asthma; atherogenesis; atheroma; atherosclerosis; atopic dermatitis; autoimmune disease; autoinflammation; bacterial infection; bacteriuria; bladder cancer; bone cancer; bone inflammation disease; brain trauma; breast cancer; bronchiolitis; bronchiolitis obliterans syndrome; cancer; cardiac infarction; cardiovascular disease; carotid artery disease; CCR2 associated neurological disorders; Cd3zeta deficiency; central nervous system disease; cerebral aneurysms; cervical cancer; chagas disease; chorioamnionitis; chronic heart failure; chronic lung disease; chronic lymphocytic leukemia; chronic myelocytic leukemia; chronic obstructive pulmonary disease (COPD); chronic respiratory viral infection; chronic urticaria; colitis; colon cancer; complex regional pain syndrome; coronary artery aneurysm; crescentic glomerulonephritis; Crohn's Disease; cystitis; cytomegalovirus retinitis; degeneration of macula and posterior pole; demyelinating disease; dengue shock syndrome; denture stomatitis; dermatosis syndrome; diabetes; diabetes mellitus, noninsulin-dependent; diabetic angiopathy; diabetic complications; diabetic macular edema; diabetic microangiopathy; diabetic nephropathy; diabetic retinitis; diabetic retinopathy; diastolic cardiomyopathies; encephalitis; endocervicitis; endometrial stromal sarcoma; endometriosis; Erdheim-Chester disease; extrapulmonary tuberculosis; extrinsic cardiomyopathy; eye disease; fibroid lung; fungal pneumonia; gingivitis; glomerulonephritis; gum disease; Hamman-Rich syndrome; head and neck cancer, herpes simplex virus keratitis; HIV-1; Hodgkin's disease; hyperhomocysteinemia; idiopathic anterior uveitis; idiopathic interstitial pneumonia; idiopathic pulmonary fibrosis; inflammation after cataract surgery; inflammatory bowel diseases; inflammatory disease; influenza; interstitial lung disease; invasive staphyloccocia; ischemia of lower members of the heart; ischemia-reperfusion injury; Israeli tick typhus; Kawasaki disease; keratitis; kidney disease; leptospirosis; limb ischemia; lipid pneumonia; lipodystrophy; lipoid nephrosis; lung cancer; lung disease; lung injury; lung transplantation; macular degeneration, age-related, 1; macular holes; malaria; malignant myeloma; mast-cell leukemia; meningitis; mesangial proliferative glomerulonephritis; metabolic disease; microvascular complications of diabetes 1; monocytic leukemia; multiple myeloma; multiple sclerosis; mycobacterium tuberculosis; myocardial infarction; myocarditis; necrosis; neovascular inflammatory disease; nephritis; nephrosclerosis; neural tube defects; neuritis; neuroinflammation; nonspecific interstitial pneumonia; obesity; ophthalmic disorder; organ allograft rejection; overnutrition; pain; pain from a sciatic nerve; papillary conjunctivitis; pelvic inflammatory disease; periodonitis; periodontal diseases; periodontitis; peripheral artery disease; peripheral pain; peritonitis; pleural tuberculosis; pleurisy; pneumoconiosis; pneumonia; post-thrombotic syndrome; primary graft dysfunction (PGD) (a reperfusion injury after transplant); proliferative glomerulonephritis; prostate cancer; psoriasis; psoriatic arthritis; pulmonary alveolar proteinosis; pulmonary fibrosis; pulmonary sarcoidosis; purulent labyrinthitis; pyelonephritis; radiculopathy; renal fibrosis; renal insufficiency; reperfusion disorders; respiratory system disease; restenosis; retinal degeneration; retinal vascular occlusion; retinal vasculitis; retinal vein occlusion; rheumatoid arthritis; rhinoscleroma; sarcoidosis; sarcoidosis 1; scleritis; secondary progressive multiple sclerosis; severe acute respiratory syndrome; silicosis; solid tumor; stachybotrys chartarum; stomach cancer; stromal keratitis; systemic lupus erythematosus; transient cerebral ischemia; transplant arteriosclerosis; trypanosomiasis; tuberculosis; tuberculous meningitis; type II diabetes; ulcerative colitis; ureteral disease; urinary system disease; urinary tract obstruction; uveitis; vangl1-related neural tube defect; vascular disease; vascular permeability and attraction of immune cells during metastasis; vasculitis; verruciform xanthoma of skin; viral infection; viral encephalitis; viral meningitis; vitreoretinopathy; or xanthogranulomatous pyelonephritis;   (ii) acute lung injury; inflammation; primary graft dysfunction (PGD); asthma; pulmonary fibrosis; COPD; atherosclerosis; lung transplant; lung injury; COPD; atherosclerosis; cancer; prostate cancer; organ transplant; metabolic disease, type II diabetes; multiple sclerosis; rheumatoid arthritis; pain; pulmonary fibrosis; or reperfusion in a lung transplant;   (iii) inflammation associated with a CCR2 associated disease, disorder, or condition; and   (iv) inflammation associated with lung injury, graft transplantation, atherosclerosis, tumor cells, or cancer.   
     
     
         19 . The method of  claim 16 , further comprising:
 (i) evaluating or monitoring a CCR2 associated disease, disorder, or condition; or   (ii) administering a CCR2 antagonist to a subject.   
     
     
         20 . A pharmaceutical composition comprising the imaging agent of  claim 1 .