Molecular probes and methods of use

Patent metadata

Field	Value
Publication number	US-10925980-B2
Application number	US-201715821943-A
Country	US
Kind code	B2
Filing date	Nov 24, 2017
Priority date	Aug 4, 2014
Publication date	Feb 23, 2021
Grant date	Feb 23, 2021

Abstract

A molecular probe includes the following formula:

P-L-C
wherein P is a EDB-FN targeting peptide, C is a contrast agent; and L is a non-peptide linker that covalently links the peptide to the contrast agent.

First claim

Having described the invention, the following is claimed:

1. A method of detecting the presence, location and/or distribution of cancer cells expressing EDB-FN comprising:
contacting a tissue of a subject with a molecular probe, the molecular probe comprising a linear peptide that includes an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9;
a contrast agent selected from at least one of a metal chelating agent or a metallofullerene; and
a non-peptide linker that covalently links the peptide to the contrast agent, wherein the linker is selected from at least one of a carboxamide and a maleimide, wherein the molecular probe targets and binds to EDB-FN in the tissue; and
detecting the molecular probe in the tissue of the subject bound to EDB-FN using an imaging modality, wherein the detected presence, location and/or distribution of the molecular probe bound to EDB-FN in the tissue is correlated with the presence, location and/or distribution of EDB-FN expressing cancer cells in the tissue.

2. The method of claim 1 , wherein the contacting step is in vivo, ex vivo, or in vitro.

3. The method of claim 1 , wherein the probe is systemically administered to a subject having or suspected of having cancer.

4. The method of claim 1 , wherein the cancer is selected from at least one of breast cancer, liver cancer, stomach cancer, colon cancer, pancreatic cancer, ovarian cancer, lung cancer, kidney cancer, prostate cancer, testicular cancer, glioblastoma, sarcoma, bone cancer, brain cancer, head and neck cancers, or skin cancer.

5. The method of claim 1 , wherein the subject is administered the probe to determine cancer aggressiveness, and wherein a greater amount of detected and/or distributed probe bound to EDB-FN compared to a control is indicative of an increase in cancer aggressiveness.

6. The method of claim 1 , wherein the non-peptide linker is a non-peptide aliphatic or heteroaliphatic linker.

7. The method of claim 1 , wherein the non-peptide linker includes an alkylene dicarboxamide when covalently linking the peptide and contrast agent.

8. The method of claim 1 , wherein the contrast agent is a magnetic resonance imaging (MRI), positron emission tomography (PET), or single photon emission computed tomography (SPECT) contrast agent.

9. The method of claim 1 , wherein the contrast agent is a metal chelating agent selected from at least one of diethylenetriaminepentaacetate (DTPA), 1,4,7,10-tetraazadodecanetetraacetate (DOTA), 1,4,7,10-tetraazadodecane-1,4,7-triacetate (DO3A), ethylenediaminetetraacetate (EDTA), 1,4,7,10-tetraazacyclotridecanetetraacetic acid (TRITA), 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid (TETA), 1,4,7,10-tetraazadodecanetetramethylacetate (DOTMA), 1,4,7,10-tetraazadodecane-1,4,7-trimethylacetate (DO3MA), N,N′,N″,N′″-tetraphosphonatomethyl-1,4,7,10-tetraazacyclododecane (DOTP), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrakis(methylene methylphosphonic acid) (DOTMP), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrakis(methylene phenylphosphonic acid) (DOTPP), or N,N′-ethylenedi-L-cysteine.

10. The method of claim 1 , the molecular probe contacted with the tissue of a subject having the formula:

wherein:
P 1 is a linear peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9;
L 2 is an amino group of the peptide P 1 , and
M is a metal selected from the group consisting of Gd +3 , Eu +3 , Tm +3 , Dy +3 , Yb +3 , Mn +2 , Fe +3 , 55 Co, 64 Cu, 67 Cu, 47 sc, 66 Ga, 68 Ga, 90 Y, 97 Ru, 99 mTC, 111 In, 109 Pd, 153 Sm, 177 Lu, 186 Re, and 188 Re, and n is an integer from 0 to 100; or salts thereof.

11. The method of claim 1 , wherein the metallofullerene is Gd3N@C80.

12. A method of detecting the presence, location and/or distribution of cancer cells expressing EDB-FN comprising:
contacting a tissue of a subject with a molecular probe having the formula:

wherein:
P 1 is a linear peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9;
L 1 is an optional spacer,
L 2 is an amino group of the peptide P 1 or the spacer, and
M is a metal selected from the group consisting of Gd +3 , Eu +3 , Tm +3 , Dy +3 , Yb +3 , Mn +2 , Fe +3 , 55 Co, 64 Cu, 67 Cu, 475 c, 66 Ga, 68 Ga, 90 Y, 97 Ru, 99 mTC, 111 In, 109 Pd, 153 Sm, 177 Lu, 186 Re, and 188 Re; or salts thereof, wherein the molecular probe targets and binds to EDB-FN in the tissue, and
detecting the molecular probes in the tissue of the subject bound to EDB-FN using an imaging modality, wherein the detected presence, location and/or distribution of the molecular probe bound to EDB-FN in the tissue is correlated with the presence, location and/or distribution of EDB-FN expressing cancer cells in the tissue.

13. The method of claim 12 , the molecular probe contacted with the tissue of a subject wherein L 1 of the probe is selected from at least one of a polyalkyleneoxide, polyvinyl alcohol, polyethylene glycol (PEG), polypropylene glycol (PPG), copolymer of ethylene and propylene glycol, polyoxyethylene (POE), polyurethane, polyphosphazene, polysaccharides, dextran, polyvinylpyrrolidones, polyvinyl ethyl ether, polyacryl amide, polyacrylate, or polycyanoacrylates.

14. The method of claim 12 , the probe being detectable upon contacting the tissue of the subject by at least one of positron emission tomography (PET) imaging and magnetic resonance imaging (MRI).

15. A method of detecting the presence, location and/or distribution of cancer cells expressing EDB-FN comprising:
contacting a tissue of a subject with a molecular probe having the formula:

wherein:
P 1 is a linear peptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9;
L 2 is an amino group of the peptide P 1 , and
M is a metal selected from the group consisting of Gd +3 , Eu +3 , Tm +3 , Dy +3 , Yb +3 , Mn +2 , Fe +3 , 55 Co, 64 Cu, 67 Cu, 47 Sc, 66 Ga, 68 Ga, 90 Y, 97 Ru, 99 mTc, 111 In, 109 Pd, 153 Sm, 177 Lu, 186 Re, and 188 Re, and
n is an integer from 0 to 100; or salts thereof, wherein the molecular probe targets and binds to EDB-FN in the tissue, and
detecting the molecular probes in the tissue of the subject bound to EDB-FN using an imaging modality, wherein the detected presence, location and/or distribution of the molecular probe bound to EDB-FN in the tissue is correlated with the presence, location and/or distribution of EDB-FN expressing cancer cells in the tissue.

16. The method of claim 15 , the probe being detectable upon contacting the tissue of the subject by at least one of positron emission tomography (PET) imaging and magnetic resonance imaging (MRI).

Assignees

• Univ Case Western Reserve

Inventors

• Lu Zheng-Rong
• Han Zheng

Classifications

• A61K51/088

  conjugates with carriers being peptides, polyamino acids or proteins (antibodies A61K51/10) · CPC title

• A61K49/14 Primary

  Peptides, e.g. proteins · CPC title

• A61K49/0032

  Methine dyes, e.g. cyanine dyes · CPC title

• A61K49/0056

  Peptides, proteins, polyamino acids · CPC title