# US 2019/0314531 A1 · Tumor and immune cell imaging based on pd-l1 expression

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- Number: US 2019/0314531 A1
- Document ID: US20190314531A1
- Status: Published application
- Kind: A1
- Application number: 16/471,678
- Filed: 2017-12-21
- Published: 2019-10-17
- Earliest claimed: 2016-12-23
- Assignees: The Johns Hopkins University
- Applicants: The Johns Hopkins University
- Inventors: Sridhar Nimmagadda; Martin Pomper; Samit Chatterjee; Wojciech Lesniak; Dhiraj Kumar
- Web page: https://patentgrep.com/patent/US20190314531A1

## CPC

- A61K 51/088: {conjugates with carriers being peptides, polyamino acids or proteins (antibodies A61K51/10)}
- A61K 49/0056: {Peptides, proteins, polyamino acids}
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 35/00: Antineoplastic agents
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

The presently disclosed subject matter provides compositions, kits, and methods comprising imaging agents that can detect Programmed Death Ligand 1 (PD-L1). The presently disclosed imaging agents can be used to detect diseases and disorders, such as cancer, infection, and inflammation, in a subject.

## Claims

### Claim 1

An imaging agent comprising a conjugate of a peptide having a binding specificity for programmed death ligand 1 (PD-L1) and a reporting moiety, and optionally a linker, wherein the linker, when present connects the peptide and the reporting moiety, and when the linker is absent, the reporting moiety is attached directly to the peptide through a primary amine of an amino acid of the peptide.

### Claim 2 (depends on 1)

The imaging agent of claim 1, wherein the peptide having a binding specificity for PD-L1 interacts with amino acids Y56, E58, A113, M115, and Y123 of PD-L1.

### Claim 3 (depends on 1, 2)

The imaging agent of claim 1 or claim 2, wherein the peptide having a binding specificity for programmed death ligand 1 (PD-L1) has at least 80% sequence identity to the peptide WL12, DK-A-221, or DK-A-222.

### Claim 4 (depends on 3)

The imaging agent of claim 3, wherein the peptide having a binding specificity for programmed death ligand 1 (PD-L1) has at least 85% sequence identity to the peptide WL12, DK-A-221, or DK-A-222.

### Claim 5 (depends on 4)

The imaging agent of claim 4, wherein the peptide having a binding specificity for programmed death ligand 1 (PD-L1) imaging agent has at least 90% sequence identity to the peptide WL12, DK-A-221, or DK-A-222.

### Claim 6 (depends on 5)

The imaging agent of claim 5, wherein the peptide having a binding specificity for programmed death ligand 1 (PD-L1) has 100% sequence identity to the peptide WL12, DK-A-221, or DK-A-222.

### Claim 7 (depends on 1)

The imaging agent of claim 1, wherein the reporting moiety is selected from the group consisting of a chelating agent, a radiolabeled substrate, a fluorescent dye, a photoacoustic reporting molecule, and a Raman-active reporting molecule.

### Claim 8 (depends on 7)

The imaging agent of claim 7, wherein the reporting moiety is a chelating agent and the chelating agent is selected from the group consisting of DOTAGA (1,4,7,10-tetraazacyclododececane,1-(glutaric acid)-4,7,10-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTASA (1,4,7,10-tetraazacyclododecane-1-(2-succinic acid)-4,7,10-triacetic acid), CB-DO2A (10-bis(carboxymethyl)-1,4,7,10-tetraazabicyclo[5.5.2]tetradecane), DEPA (7-[2-(Bis-carboxymethylamino)-ethyl]-4,10-bis-carboxymethyl-1,4,7,10-tetraaza-cyclododec-1-yl-acetic acid)), 3p-C-DEPA (2-[(carboxymethyl)][5-(4-nitrophenyl-1-[4,7,10-tris (carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]pentan-2-yl)amino]acetic acid)), TCMC (2-(4-isothiocyanotobenzyl)-1,4,7,10-tetraaza-1,4,7,10-tetra-(2-carbamonyl methyl)-cyclododecane), oxo-DO3A (1-oxa-4,7,10-triazacyclododecane-5-S-(4-isothiocyanatobenzyl)-4,7,10-triacetic acid), p-NH2-Bn-Oxo-DO3A (1-Oxa-4,7,10-tetraazacyclododecane-5-S-(4-aminobenzyl)-4,7,10-triacetic acid), TE2A ((1,8-N,N-bis-(carboxymethyl)-1,4,8,11-tetraazacyclotetradecane), MM-TE2A, DM-TE2A, CB-TE2A (4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane), CB-TE1A1P (4,8,11-tetraazacyclotetradecane-1-(methanephosphonic acid)-8-(methanecarboxylic acid), CB-TE2P (1,4,8,11-tetraazacyclotetradecane-1,8-bis(methanephosphonic acid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), NOTA (1,4,7-triazacyclononane-N,N′,N″-triacetic acid), NODA (1,4,7-triazacyclononane-1,4-diacetate); NODAGA (1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid), (NOTAGA) 1,4,7-triazonane-1,4-diyl)diacetic acid DFO (Desferoxamine), NETA ([4-[2-(bis-carboxymethylamino)-ethyl]-7-carboxymethl-[1,4,7]triazonan-1-yl}-acetic acid), TACN-TM (N,N′,N″, tris(2-mercaptoethyl)-1,4,7-triazacyclononane), Diamsar (1,8-Diamino-3,6,10,13,16,19-hexaazabicyclo(6,6,6)eicosane, 3,6,10,13,16,19-Hexaazabicyclo[6.6.6]eicosane-1,8-diamine), Sarar (1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexaazabicyclo[6.6.6] eicosane-1,8-diamine), AmBaSar (4-((8-amino-3,6,10,13,16,19-hexaazabicyclo [6.6.6] icosane-1-ylamino) methyl) benzoic acid), and BaBaSar.

### Claim 9 (depends on 7)

The imaging agent of claim 7, wherein the chelating agent is selected from the group consisting of:

### Claim 10 (depends on 7)

The imaging agent of claim 7, wherein the reporting moiety is a chelating agent and the chelating agent further comprises a radiometal selected from the group consisting of 94mTc, 99mTc, 111In, 67Ga, 68Ga, 86Y, 90Y, 177Lu, 186Re, 188Re, 60Cu, 61Cu, 62Cu, 64Cu, 67Cu, 55Co, 57Co, 47Sc, 225Ac, 213Bi, 212Bi, 212Pb, 153Sm, 166Ho, 152Gd, 82Rb, 89Zr, and 166Dy.

### Claim 11 (depends on 7)

The imaging agent of claim 7, wherein the reporting moiety is a radiolabeled substrate and the radiolabeled substrate comprises a radioisotope selected from the group consisting of 11C, 13N, 15O, 123I, 124I, 125I, 126I, 131I, 75Br, 76Br, 77Br, 80Br, 80mBr, 82Br, 83Br, 18F, and 211At.

### Claim 12 (depends on 11)

The imaging agent of claim 11, wherein the radiolabeled substrate comprises an 18F-labeled substrate.

### Claim 13 (depends on 12)

The imaging agent of claim 12, wherein the 18F-labeled substrate is selected from the group consisting of 2-fluoro-PABA, 3-fluoro-PABA, 2-fluoro-mannitol, and N-succinimidyl-4-fluorobenzoate.

### Claim 14 (depends on 1)

The imaging agent of claim 1, wherein the reporting moiety is directly incorporated into the peptide.

### Claim 15 (depends on 14)

The imaging agent of claim 14, wherein the reporting moiety comprises a radiolabeled amino acid of the peptide.

### Claim 16 (depends on 15)

The imaging agent of claim 15, wherein the radiolabeled amino acid is selected from the group consisting of iodotyrosine and fluorotyrosine.

### Claim 17 (depends on 7)

The imaging agent of claim 7, wherein the reporting moiety is a fluorescent dye and the fluorescent dye is selected from the group consisting of: carbocyanine, indocarbocyanine, oxacarbocyanine, thuicarbocyanine, merocyanine, polymethine, coumarine, rhodamine, xanthene, fluorescein, a boron-dipyrromethane (BODIPY) dye, Cy5, Cy5.5, Cy7, VivoTag-680, VivoTag-S680, VivoTag-S750, AlexaFluor660, AlexaFluor680, AlexaFluor700, AlexaFluor750, AlexaFluor790, Dy677, Dy676, Dy682, Dy752, Dy780, DyLight547, Dylight647, HiLyte Fluor 647, HiLyte Fluor 680, HiLyte Fluor 750, IR Dye 800, IRDye 800CW, IRDye 800RS, IRDye 700DX, ADS780WS, ADS830WS, and ADS832WS.

### Claim 18 (depends on 7)

The imaging agent of claim 7, wherein the reporting moiety is a photoacoustic reporting molecule and the photoacoustic reporting molecule is selected from the group consisting of a dye or a nanoparticle.

### Claim 19 (depends on 18)

The imaging agent of claim 18, wherein the dye comprises a fluorescent dye.

### Claim 20 (depends on 19)

The imaging agent of claim 19, wherein the fluorescent dye is selected from the group consisting of indocyanine-green (ICG), Alexa Fluor 750, Evans Blue, BHQ3, QXL680, IRDye880CW, MMPSense 680, Methylene Blue, PPCy-C8, and Cypate-C 18.

### Claim 21 (depends on 18)

The imaging agent of claim 18, wherein the nanoparticle is selected from the group consisting of a plasmonic nanoparticle, a quantum dot, a nanodiamond, a polypyrrole nanoparticle, a copper sulfide nanoparticle, a graphene nanosheet, an iron oxide-gold core-shell nanoparticle, a Gd2O3 nanoparticle, a single-walled carbon nanotube, a dye-loaded perfluorocarbon nanoparticle, and a superparamagnetic iron oxide nanoparticle.

### Claim 22 (depends on 7)

The imaging agent of claim 7, wherein the reporting moiety is a Raman-active reporting molecule and the Raman-active reporting molecule is selected from the group consisting of a single-walled carbon nanotube (SWNT) and a surface-enhanced Raman scattering (SERS) agent.

### Claim 23 (depends on 22)

The imaging agent of claim 22, wherein the SERS agent comprises a metal nanoparticle labeled with a Raman-active reporter molecule.

### Claim 24 (depends on 23)

The imaging agent of claim 23, wherein the Raman-active reporter molecule comprises a fluorescent dye.

### Claim 25 (depends on 24)

The imaging agent of claim 24, wherein the fluorescent dye is selected from the group consisting of Cy3, Cy5, rhodamine, and a chalcogenopyrylium dye.

### Claim 26 (depends on 7)

The imaging agent of claim 7, wherein the linker is selected from the group consisting of:
(a)
wherein:
Rpt is the reporting moiety;
W1 is selected from the group consisting of C1-C6 alkylene, C3-C6 cycloalkylene, and arylene;
W2 is selected from the group consisting of —NR1—(C═O)—, —NR1—(C═S)—, —(C═O)—NR1—, —(C═S)—NR1—, and —S—, wherein each R1 is independently H or C1-C4 alkyl;
each R2 is independently H or —COOR3, wherein each R3 is independently H, C1-C6 alkyl, C2-C12 aryl or C4-C16 alkylaryl;
b is an integer selected from the group consisting of 0, 1, 2, and 3;
d is an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, and 8; and
wherein the wavy line indicates a point of attachment between the linker and the peptide;
(b) Rpt-X—Y—Z—W3—
wherein:
Rpt is the reporting moiety;
X and Z are each independently C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C1-C8 heteroalkyl, C2-C8 heteroalkenyl, C2-C8 heteroalkynyl, C1-C8 alkoxy, or a bond, each of which may be substituted with 0-5 RA;
Y and W3 are each independently —O—, —S(O)p—, —NH—, —NRB—, —CH═CH—, —CRB═CH—, —CH═CRB—, —NH—CO—, —NH—CO2—, —NRB—CO—, —NRB—CO2—; —CO—NH—, —CO2—NH—, —CO—NRB—, —CO2—NRB—, or a bond;
p is 0, 1, or 2;
RA, for each occurrence, is halogen, hydroxy, amino, cyano, nitro, CO2H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclo, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted mono or dialkylamino, optionally substituted alkylthio, optionally substituted alkylsulfinyl, optionally substituted alkylsulfonyl, optionally substituted mono- or dialkylcarboxamide, optionally substituted aryl, or optionally substituted heteroaryl; and RB, for each occurrence, is optionally substituted alkyl, optionally substituted alkoxy, optionally substituted mono or dialkylamino, optionally substituted alkylthio, optionally substituted aryl, or optionally substituted heteroaryl; or
(c) an amino acid linker.

### Claim 27 (depends on 1)

The imaging agent of claim 1, wherein the imaging agent is selected from the group consisting of a compound of formula (I), formula (II), or formula (III):
wherein:
n is an integer selected from the group consisting of 0 and 1;
L is a linker; and
Rpt is a reporting moiety; and
wherein the reporting moiety or linker, when present, is attached to a primary amine group of an amino acid of the peptide comprising the imaging agent of formula (I), formula (II), or formula (III).

### Claim 27 (depends on 26)

The imaging agent of claim 26, wherein the linker, when present, is attached to an 13ornithine (Orn) primary amine group of the compound of formula (I).

### Claim 28 (depends on 26)

The imaging agent of claim 26, wherein the reporting moiety comprises a DOTAGA chelating agent.

### Claim 29 (depends on 28)

The imaging agent of claim 28, wherein the DOTAGA chelating agent further comprises a 64Cu radiometal.

### Claim 30 (depends on 26)

The imaging agent of claim 26, wherein the compound of formula (I) is:

### Claim 31 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)

An imaging method for detecting Programmed Death Ligand 1 (PD-L1), the method comprising:
(a) providing an effective amount of an imaging agent of any of claims 1-27;
(b) contacting one or more cells or tissues with the imaging agent; and
(c) making an image to detect PD-L1.

### Claim 32 (depends on 31)

The imaging method of claim 31, wherein contacting of the one or more cells or tissues with the imaging agent is performed in vitro, in vivo, or ex vivo.

### Claim 33 (depends on 32)

The imaging method of claim 32, wherein contacting of the one or more cells or tissues with the imaging agent is performed in a subject.

### Claim 34 (depends on 33)

The imaging method of claim 33, wherein the subject is a human, rat, mouse, cat, dog, horse, sheep, cow, monkey, avian, or amphibian.

### Claim 35 (depends on 31)

The imaging method of claim 31, wherein detection of the PD-L1 occurs at about 60-120 minutes or less after administration of the imaging agent to the subject.

### Claim 36 (depends on 31)

The imaging method of claim 31, wherein the imaging method is used to detect a cancer.

### Claim 37 (depends on 36)

The imaging agent of claim 36, wherein the cancer is selected from the group consisting of a blastoma, a carcinoma, a glioma, a leukemia, a lymphoma, a melanoma, a myeloma, a sarcoma, head cancer, neck cancer, head and neck cancer, lung cancer, breast cancer, triple negative breast cancer, prostate cancer, colorectal cancer, esophageal cancer, stomach cancer, leukemia/lymphoma, uterine cancer, skin cancer, endocrine cancer, urinary cancer, pancreatic cancer, gastrointestinal cancer, ovarian cancer, cervical cancer, renal cancer, bladder cancer, brain cancer, adenoma, and a metastatic cancer.

### Claim 38 (depends on 31)

The imaging method of claim 31, wherein the imaging method is used to detect a solid tumor.

### Claim 39 (depends on 38)

The imaging method of claim 38, wherein the solid tumor is in an organ selected from the group consisting of brain, colon, breast, prostate, liver, kidney, lung, esophagus, head and neck, ovary, cervix, stomach, rectum, bladder, uterus, testes, and pancreas.

### Claim 40 (depends on 31)

The imaging method of claim 31, wherein the imaging method is used to detect an infection.

### Claim 41 (depends on 40)

The imaging method of claim 40, wherein the infection is a microbial infection.

### Claim 42 (depends on 41)

The imaging method of claim 41, wherein the microbial infection is selected from the group consisting of an infection due to one or more microorganisms selected from the group consisting of Mycobacterium tuberculosis, E. coli, Klebsiella sp., Enterobacter sp., Proteus sp., Serratia marcescens, Pseudomonas aeruginosa, Staphylococcus spp., including S. aureus and coag.-negative Staphylococcus, Enterococcus sp., Streptococcus pneumoniae, Haemophilus influenzae, Bacteroides spp., Acinetobacter spp., Helicobacter spp., Candida sp., methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VRE).

### Claim 43 (depends on 31)

The imaging method of claim 31, wherein the imaging method is used to detect inflammation.

### Claim 44 (depends on 43)

The imaging agent of claim 43, when in the inflammation is related to a disorder selected from the group consisting of asthma, an autoimmune disease, an autoinflammatory disease, Celiac disease, diverticulitis, glomerulonephritis, hidradenitis suppurativa, a hypersensitivity, an inflammatory bowel disease, interstitial cystitis, otitis, pelvic inflammatory disease, reperfusion injury, rheumatic fever, rheumatoid arthritis, sarcoidosis, transplant rejection, lupus, systemic lupus erythematosus, and vasculitis.

### Claim 45 (depends on 44)

The imaging method of claim 44, wherein the inflammation is caused by rheumatoid arthritis or systemic lupus erythematosus.

### Claim 46 (depends on 31)

The imaging method of claim 31, wherein the imaging method is used to detect one or more immune cells in a tumor.

### Claim 47 (depends on 31)

The imaging method of claim 31, wherein the imaging method is used to detect systemic distribution of immune cells in the tumor or in a subject.

### Claim 48 (depends on 31)

The imaging method of claim 31, wherein the imaging method is used to detect an immune cell response to an infectious disease.

### Claim 49 (depends on 31)

The imaging method of claim 31, wherein the imaging method is used to detect an immune cell response in a tumor or in normal tissue response to an inflammatory disease.

### Claim 50 (depends on 31)

The imaging method of claim 31, wherein the imaging method detects PD-L1 expression levels in the subject.

### Claim 51 (depends on 31)

The imaging method of claim 31, wherein the imaging method measures an occupancy or target engagement by antibodies or peptides or low molecular weight agents of PD-L1 at a tumor site or in normal tissue of the subject.

### Claim 52 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)

A kit for detecting Programmed Death Ligand 1 (PD-L1), the kit comprising the imaging agent of any of claims 1-27.