Methods and Compositions for Improved F-18 Labeling of Proteins, Peptides and Other Molecules

Abstract

The present application discloses compositions and methods of synthesis and use of 18 F- or 19 F-labeled molecules of use in PET, SPECT and/or MR imaging. Preferably, the 18 F or 19 F is conjugated to a targeting molecule by formation of a complex with a group IIIA metal and binding of the complex to a bifunctional chelating agent, which may then be directly or indirectly attached to the targeting molecule. In other embodiments, the 18 F or 19 F labeled moiety may comprise a targetable construct used in combination with a bispecific antibody to target a disease-associated antigen. The disclosed methods and compositions allow the simple and reproducible labeling of molecules at very high efficiency and specific activity in 30 minutes or less. In preferred embodiments, the bifunctional chelating agent bound to 18 F- or 19 F-metal complex may be conjugated to the molecule to be labeled at a reduced temperature, e.g. room temperature.

What is claimed is: 
     
         1 . A method of imaging comprising:
 a) labeling a bifunctional chelating agent with a complex of (i)  18 F or  19 F, and (ii) a group IIIA metal;   b) attaching the  18 F or  19 F labeled bifunctional chelating agent to a molecule to form an  18 F- or  19 F-labeled molecule, wherein the molecule is selected from the group consisting of an antibody, antigen-binding antibody fragment, bispecific antibody, affibody, diabody, minibody, scFv, aptamer, avimer, targeting peptide, somatostatin, bombesin, octreotide, RGD peptide and folate;   c) administering the  18 F- or  19 F-labeled molecule to a subject; and   d) imaging the distribution of the  18 F- or  19 F-labeled molecule by positron emission tomography (PET), magnetic resonance imaging (MM) or single photon emission computer tomography (SPECT).   
     
     
         2 . The method of  claim 1 , wherein the  18 F-metal or  19 F-metal complex is attached to the bifunctional chelating agent at a temperature between 90° C. and 110° C. 
     
     
         3 . The method of  claim 1 , wherein the bifunctional chelating agent is attached to the molecule at room temperature. 
     
     
         4 . The method of  claim 1 , wherein the bifunctional chelating agent is selected from the group consisting of NODA-BA, NODA-BAEM, NODA-BM, NODA-butyne, NODA-EA, NODA-EBA, NODA-EPA, NODA-EPN, NODA-HA, NODA-MBA, NODA-MBEM, NODA-MPAA, NODA-MPAA NHS ester, NDOA-MPAEM, NODA-MPAPEG.sub.3M. sub.3, NODA-MPH, NODA-MPN, NODA-2-nitroimidazole, NODA-PA, NODA-PAEM and NODA-propyl amine. 
     
     
         6 . The method of  claim 1 , wherein the group IIIA metal is aluminum. 
     
     
         7 . The method of  claim 1 , wherein a metal- 18 F or metal- 19 F complex is attached to the bifunctional chelating agent by heating in an aqueous medium at a temperature between 50° C. and 110° C. 
     
     
         8 . The method of  claim 1 , wherein the efficiency of labeling with  18 F is at least 35% at 50° C. 
     
     
         9 . The method of  claim 1 , wherein the specific activity of the labeled molecule is at least 4,000 Ci/mmol. 
     
     
         10 . The method of  claim 1 , wherein a metal- 18 F or metal- 19 F complex is attached to the bifunctional chelating agent in an aqueous medium comprising aluminum, trehalose, potassium biphthalate, ethanol and ascorbic acid at a pH between 3.9 and 4.2. 
     
     
         11 . The method of  claim 1 , wherein multiple copies of the bifunctional chelating agent are attached to the molecule. 
     
     
         12 . The method of  claim 1 , wherein the  18 F- or  19 F-labeled molecule is produced in less than 30 minutes from the start of the method. 
     
     
         13 . The method of  claim 1 , wherein the bifunctional chelating agent is attached to the molecule by a click chemistry reaction or by a maleimide-sulfhydryl reaction. 
     
     
         14 . The method of  claim 1 , wherein the yield of labeled molecule is at least 95%. 
     
     
         15 . The method of  claim 11 , further comprising: e) analyzing the distribution of the labeled molecule to detect, diagnose or image the presence of a disease in the subject, wherein the disease is selected from the group consisting of cancer, a cardiovascular disease, an infectious disease, an inflammatory disease, an autoimmune disease, an immune dysfunction disease, graft versus host disease, organ transplant rejection and a neurological disease. 
     
     
         16 . The method of  claim 15 , wherein the labeled molecule binds to an antigen selected from the group consisting of carbonic anhydrase IX, CCL19, CCL21, CSAp, CD1, CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, IGF-1R, CD20, CD21, CD22, CD23, CD25, CD29, CD30, CD32b, CD33, CD37, CD38, CD40, CD40L, CD45, CD46, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD70, CD74, CD79a, CD80, CD83, CD95, CD126, CD133, CD138, CD147, CD154, CXCR4, CXCR7, CXCL12, HIF-1α, AFP, PSMA, CEACAMS, CEACAM-6, c-met, B7, ED-B of fibronectin, Factor H, Flt-1, Flt-3, folate receptor, GRO-β, HMGB-1, hypoxia inducible factor (HIF), HM1.24, insulin-like growth factor-1 (ILGF-1), IFN-γ, IFN-α, IFN-β, IL-2, IL-4R, IL-6R, IL-13R, IL-15R, IL-17R, IL-18R, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-25, IP-10, MAGE, mCRP, MCP-1, MIP-1A, MIP-1B, MIF, MUC1, MUC2, MUC3, MUC4, MUC5, NCA-95, NCA-90, Ia, HM1.24, EGP-1, EGP-2, HLA-DR, tenascin, Le(y), RANTES, T101, TAC, Tn antigen, Thomson-Friedenreich antigens, tumor necrosis antigens, TNF-α, TRAIL receptor R1, TRAIL receptor R2, VEGFR, EGFR, P1GF, complement factors C3, C3a, C3b, C5a, and C5. 
     
     
         17 . The method of  claim 15 , wherein the labeled molecule is an antibody selected from the group consisting of hR1 (anti-IGF-1R), hPAM4 (anti-pancreatic cancer mucin), hA20 (anti-CD20), hA19 (anti-CD19), hIMMU31 (anti-AFP), hLL1 (anti-CD74), hLL2 (anti-CD22), hMu-9 (anti-CSAp), hL243 (anti-HLA-DR), hMN-14 (anti-CEACAM5), hMN-15 (anti-CEACAM6), hRS7 (anti-EGP-1) and hMN-3 (anti-CEACAM6). 
     
     
         18 . The method of  claim 1 , wherein the bifunctional chelating agent is conjugated to a molecule via an amide, ester, anhydride, carbonate, carbamate, dithiocarbamate, ether, thioether, disulfide, urea, thiourea, triazoyl, amine, imine, oxime or hydrazone bond. 
     
     
         19 . The method of  claim 1 , wherein the  18 F-metal or  19 F-metal complex is attached to the bifunctional chelating agent in the presence of an organic solvent selected from the group consisting of ethanol, acetonitrile, MeCN, DMF and THF. 
     
     
         20 . The method of  claim 1 , wherein the molecule is selected from the group consisting of IMP449, IMP460, IMP461, IMP467, IMP469, IMP470, IMP471, IMP479, IMP485, IMP486, IMP487, IMP488, IMP490, IMP493, IMP495, IMP497, IMP500, IMP508 and IMP517.