# US 2017/0283442 A1 · Methods and Compositions for Improved F-18 Labeling of Proteins, Peptides and Other Molecules

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- Number: US 2017/0283442 A1
- Document ID: US20170283442A1
- Status: Published application
- Kind: A1
- Application number: 15/621,571
- Filed: 2017-06-13
- Published: 2017-10-05
- Earliest claimed: 2007-01-11
- Assignees: Immunomedics, Inc.
- Applicants: Immunomedics, Inc.
- Inventors: Christopher A. D'Souza; William J. McBride; David M. Goldenberg
- Web page: https://patentgrep.com/patent/US20170283442A1

## CPC

- C07F 5/069: {without C-aluminium linkages}
- C07B 59/002: {Heterocyclic compounds}
- C07D 207/452: with hydrocarbon radicals, substituted by hetero atoms, directly attached to the ring nitrogen atom
- C07D 255/02: not condensed with other rings
- C07D 403/12: linked by a chain containing hetero atoms as chain links
- C07D 405/12: linked by a chain containing hetero atoms as chain links
- C07D 475/04: with a nitrogen atom directly attached in position 2
- C07K 7/06: having 5 to 11 amino acids
- C07B 2200/05: Isotopically modified compounds, e.g. labelled
- C07F: ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM (metal-containing porphyrins C07D487/22)
- C07B: GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR (preparation of carboxylic acid esters by telomerisation C07C67/47; telomerisation C08F)
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

The present application discloses compositions and methods of synthesis and use of 18F- or 19F-labeled molecules of use in PET, SPECT and/or MR imaging. Preferably, the 18F or 19F 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 18F or 19F 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 18F- or 19F-metal complex may be conjugated to the molecule to be labeled at a reduced temperature, e.g. room temperature.

## Claims

### Claim 1

A method of imaging comprising:
a) labeling a bifunctional chelating agent with a complex of (i) 18F or 19F, and (ii) a group IIIA metal;
b) attaching the 18F or 19F labeled bifunctional chelating agent to a molecule to form an 18F- or 19F-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 18F- or 19F-labeled molecule to a subject; and
d) imaging the distribution of the 18F- or 19F-labeled molecule by positron emission tomography (PET), magnetic resonance imaging (MM) or single photon emission computer tomography (SPECT).

### Claim 2 (depends on 1)

The method of claim 1, wherein the 18F-metal or 19F-metal complex is attached to the bifunctional chelating agent at a temperature between 90° C. and 110° C.

### Claim 3 (depends on 1)

The method of claim 1, wherein the bifunctional chelating agent is attached to the molecule at room temperature.

### Claim 4 (depends on 1)

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.

### Claim 6 (depends on 1)

The method of claim 1, wherein the group IIIA metal is aluminum.

### Claim 7 (depends on 1)

The method of claim 1, wherein a metal-18F or metal-19F complex is attached to the bifunctional chelating agent by heating in an aqueous medium at a temperature between 50° C. and 110° C.

### Claim 8 (depends on 1)

The method of claim 1, wherein the efficiency of labeling with 18F is at least 35% at 50° C.

### Claim 9 (depends on 1)

The method of claim 1, wherein the specific activity of the labeled molecule is at least 4,000 Ci/mmol.

### Claim 10 (depends on 1)

The method of claim 1, wherein a metal-18F or metal-19F 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.

### Claim 11 (depends on 1)

The method of claim 1, wherein multiple copies of the bifunctional chelating agent are attached to the molecule.

### Claim 12 (depends on 1)

The method of claim 1, wherein the 18F- or 19F-labeled molecule is produced in less than 30 minutes from the start of the method.

### Claim 13 (depends on 1)

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.

### Claim 14 (depends on 1)

The method of claim 1, wherein the yield of labeled molecule is at least 95%.

### Claim 15 (depends on 11)

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.

### Claim 16 (depends on 15)

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.

### Claim 17 (depends on 15)

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).

### Claim 18 (depends on 1)

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.

### Claim 19 (depends on 1)

The method of claim 1, wherein the 18F-metal or 19F-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.

### Claim 20 (depends on 1)

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.