# US 11,442,063 B2 · Broad spectrum kinase binding agents

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- Number: US 11,442,063 B2
- Document ID: US11442063B2
- Status: Granted
- Kind: B2
- Application number: 16/426,982
- Filed: 2019-05-30
- Published: 2020-01-02
- Granted: 2022-09-13
- Earliest claimed: 2018-05-30
- Assignees: Promega Corporation
- Applicants: Promega Corporation
- Inventors: Cesear Corona; Poncho Meisenheimer; Matthew Robers; James Vasta
- Web page: https://patentgrep.com/patent/US11442063B2

## CPC

- G01N 33/54353: {with ligand attached to the carrier via a chemical coupling agent (coatings G01N33/54393)}
- C07D 239/48: Two nitrogen atoms
- C07F 5/022: {without C-boron linkages}
- G01N 1/34: Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)}
- G01N 33/573: for enzymes or isoenzymes
- G01N 21/6428: {Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53)}
- G01N 2021/6439: {with indicators, stains, dyes, tags, labels, marks}
- G01N 2333/91205: Phosphotransferases in general
- G01R 33/5601: {involving use of a contrast agent for contrast manipulation, e.g. a paramagnetic, super-paramagnetic, ferromagnetic or hyperpolarised contrast agent}
- G01T 1/1642: {using a scintillation crystal and position sensing photodetector arrays, e.g. ANGER cameras}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- C07F: ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM (metal-containing porphyrins C07D487/22)
- G01R: MEASURING ELECTRIC VARIABLES; MEASURING MAGNETIC VARIABLES (indicating correct tuning of resonant circuits H03J3/12)
- G01T: MEASUREMENT OF NUCLEAR OR X-RADIATION (radiation analysis of materials, mass spectrometry G01N23/00; tubes for determining the presence, intensity, density or energy of radiation or particles H01J47/00)

## Abstract

Provided herein are compounds that are broad-spectrum protein kinase binding agents, detectable tracers comprising such compounds, and method of use thereof for the detection of protein kinases.

## Claims

### Claim 1

A broad-spectrum kinase binding agent of formula:

### Claim 2 (depends on 1)

The broad-spectrum kinase binding agent of claim 1, comprising a non-natural abundance of one or more stable heavy isotopes.

### Claim 3

A broad-spectrum kinase binding agent attached to a functional element or solid surface, comprising a moiety of formula:
attached to the functional element or solid surface.

### Claim 4 (depends on 3)

The broad-spectrum kinase binding agent of claim 3, wherein the solid surface is selected from a sedimental particle, a membrane, glass, a tube, a well, a self-assembled monolayer, a surface plasmon resonance chip, or a solid support with an electron conducting surface.

### Claim 5 (depends on 4)

The broad-spectrum kinase binding agent of claim 4, wherein the sedimental particle is a magnetic particle.

### Claim 6 (depends on 3)

The broad-spectrum kinase binding agent of claim 3, wherein the functional element is selected from a detectable element, an affinity element, and a capture element.

### Claim 7 (depends on 6)

The broad-spectrum kinase binding agent of claim 6, wherein the detectable element comprises a fluorophore, chromophore, radionuclide, electron opaque molecule, a MRI contrast agent, SPECT contrast agent, or mass tag.

### Claim 8 (depends on 3)

The broad-spectrum kinase binding agent of claim 3, wherein CC-1852 or CC-1861 is attached to the functional element directly or via a linker.

### Claim 9 (depends on 8)

The broad-spectrum kinase binding agent of claim 8, wherein the linker comprises —[(CH2)2O]n—, wherein n is 1-20.

### Claim 10 (depends on 9)

The broad-spectrum kinase binding agent of claim 9, wherein the linker is attached to CC-1852 or CC-1861, and/or the functional element, by an amide bond.

### Claim 11 (depends on 3)

The broad-spectrum kinase binding agent of claim 3, comprising a structure of:
wherein X is a functional element or solid surface.

### Claim 12 (depends on 11)

The broad-spectrum kinase binding agent of claim 11, wherein X is a fluorophore.

### Claim 13 (depends on 12)

The broad-spectrum kinase binding agent of claim 12, comprising a structure of:

### Claim 14 (depends on 3)

The broad-spectrum kinase binding agent of claim 3, comprising a structure of:
wherein X is a functional element or solid surface.

### Claim 15 (depends on 14)

The broad-spectrum kinase binding agent of claim 14, wherein X is a fluorophore.

### Claim 16 (depends on 15)

The broad-spectrum kinase binding agent of claim 15, comprising a structure of:

### Claim 17 (depends on 3)

A method of detecting or quantifying kinases in a sample, comprising contacting the sample with a composition of claim 3 and detecting or quantifying the functional element of a signal produced thereby.

### Claim 18 (depends on 3)

A method of isolating kinases from a sample, comprising contacting the sample with a broad-spectrum kinase binding agent of claim 3, and separating the functional element of the solid surface, as well as the bound kinases, from the unbound portion of the sample.

### Claim 19 (depends on 18)

A method of characterizing the identities of the kinases in a sample comprising isolating the kinases from a sample by the method of claim 18, and analyzing the isolated kinases by mass spectrometry.

### Claim 20 (depends on 3)

A method of monitoring interactions between kinases and unmodified biomolecules comprising contacting the sample with a broad-spectrum kinase binding agent of claim 3.