# US 12,215,324 B2 · Monocyte and macrophage binding aptamers and their application

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 12,215,324 B2
- Document ID: US12215324B2
- Status: Granted
- Kind: B2
- Application number: 17/849,513
- Filed: 2022-06-24
- Published: 2023-01-19
- Granted: 2025-02-04
- Earliest claimed: 2021-06-24
- Assignees: University of Washington; Seattle Children's Hospital
- Applicants: University of Washington; Seattle Children's Hospital
- Inventors: Suzie Hwang Pun; Meilyn Sylvestre; Nataly Kacherovsky; Emmeline Cheng; Ian Cardle; Chris Saxby
- Web page: https://patentgrep.com/patent/US12215324B2

## CPC

- C12N 15/115: Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith {; Nucleic acids binding to non-nucleic acids, e.g. aptamers}
- A61K 39/39: characterised by the immunostimulating additives, e.g. chemical adjuvants
- A61K 40/17: Monocytes; Macrophages
- A61K 40/24: Antigen-presenting cells [APC]
- A61K 40/42: Cancer antigens
- C12N 5/0645: {Macrophages, e.g. Kuepfer cells in the liver; Monocytes}
- C12N 15/1048: {SELEX}
- C12N 15/1065: {Preparation or screening of tagged libraries, e.g. tagged microorganisms by STM-mutagenesis, tagged polynucleotides, gene tags}
- G01N 33/5308: {for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites}
- G01N 33/56972: {White blood cells}
- A61K 2239/31: characterized by the route of administration
- A61K 2239/38: characterised by the dose, timing or administration schedule
- C12N 2310/16: Aptamers
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)
- 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)

## Abstract

Provided herein are compositions comprising aptamers that specifically bind monocytes and/or macrophage and methods for their use. These aptamer compositions can be used in methods for isolating and/or enriching monocytes and/or macrophages or depleting cell populations of monocytes and/or macrophages. Further provided are methods of using the aptamers or cell populations generated using them in the methods disclosed herein for therapies and/or drug delivery.

## Claims

### Claim 1

A composition comprising an aptamer that specifically binds to a monocyte and/or a macrophage, wherein the aptamer comprises a sequence having at least 75% sequence identity to SEQ ID NO:1; and wherein the aptamer can further comprise a number (N) of nucleotides (nt) at each end wherein each nucleotide is selected independently, and wherein each N comprises from 3 nt to 30 nt, from 3 nt to 20 nt, or from 3 nt to 10 nt;
wherein any nucleotide change in the sequence of the aptamer relative to SEQ ID NO:1 occurs within one or more stem regions of SEQ ID NO:1 and co-occurs with one or more compensatory nucleotide changes within the one or more stem regions of SEQ ID NO:1 to maintain double-strandedness of the one or more stem regions of SEQ ID NO:1.

### Claim 2 (depends on 1)

The composition of claim 1, wherein the aptamer comprises a sequence having at least 75% identity to SEQ ID NO:2 and any nucleotide change in the sequence of the aptamer relative to SEQ ID NO:2 occurs within one or more stem regions of SEQ ID NO:2 and co-occurs with one or more compensatory nucleotide changes within the one or more stem regions of SEQ ID NO:2 to maintain double-strandedness of the one or more stem regions of SEQ ID NO: 2; or
wherein the aptamer comprises a sequence having at least 75% identity to SEQ ID NO:3 and any nucleotide change in the sequence of the aptamer relative to SEQ ID NO:3 occurs within one or more stem regions of SEQ ID NO:3 and co-occurs with one or more compensatory nucleotide changes within the one or more stem regions of SEQ ID NO:3 to maintain double-strandedness of the one or more stem regions of SEQ ID NO:3.

### Claim 3 (depends on 1)

The composition of claim 1, wherein the monocyte and/or macrophage expresses CD14.

### Claim 4 (depends on 1)

The composition of claim 1, wherein the macrophage is an M0 or M2 macrophage.

### Claim 5 (depends on 1)

The composition of claim 1, wherein the aptamer comprises at least five single-stranded loops and at least four double-stranded stems.

### Claim 6 (depends on 1)

The composition of claim 1, wherein the aptamer comprises the sequence of SEQ ID NO:1.

### Claim 7 (depends on 1)

The composition of claim 1, wherein the aptamer is attached to a solid support or phase-changing agent.

### Claim 8 (depends on 1)

The composition of claim 1, wherein the aptamer further comprises a detectable moiety, a label, a tag, or a probe.

### Claim 9 (depends on 1)

The composition of claim 1, formulated for use as a drug delivery device, or for use as a sensor.

### Claim 10

A cell displaying an aptamer that specifically binds to a monocyte and/or a macrophage, wherein the aptamer comprises a sequence having at least 75% sequence identity, or differs by less than three nucleotides, to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3;
wherein any nucleotide change in the sequence of the aptamer relative to SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO:3 occurs within one or more stem regions of SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO:3 and co-occurs with one or more compensatory nucleotide changes within the one or more stem regions of SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO:3 to maintain double-strandedness of the one or more stem regions of SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO: 3.

### Claim 11 (depends on 1)

A method for isolating or enriching monocytes and macrophages from a biological sample comprising a plurality of cell types, the method comprising:
(i) contacting the biological sample with an aptamer of claim 1 that specifically binds a monocyte and/or macrophage under conditions that permit forming aptamer-bound cells; and
(ii) separating the aptamer-bound cells from cells not bound to the aptamer.

### Claim 12 (depends on 11)

The method of claim 11, wherein the biological sample comprises a blood sample or a processed blood sample.

### Claim 13 (depends on 11)

The method of claim 11, wherein the aptamer is attached to a solid support or phase-changing agent.

### Claim 14 (depends on 11)

The method of claim 11, wherein the aptamer further comprises a detectable moiety, a label, a tag, or a probe.

### Claim 15 (depends on 1)

A method for depleting monocytes and macrophages from a biological sample comprising a plurality of cell types, the method comprising:
(i) contacting the biological sample with an aptamer of claim 1 that specifically binds a monocyte and/or macrophage under conditions that permit forming an aptamer-bound cell;
(ii) separating the aptamer-bound cells from cells not bound to the aptamer; and
(iii) recovering the cells not bound to the aptamer by removing aptamer-bound cells,
whereby the monocytes and/or macrophages are depleted from the biological sample.

### Claim 16 (depends on 15)

The method of claim 15, wherein the biological sample comprises a blood sample or a processed blood sample.

### Claim 17 (depends on 1)

A pharmaceutical composition comprising an aptamer of claim 1.

### Claim 18 (depends on 11)

A method of treating a disease, the method comprising: administering the isolated or enriched monocytes and/or macrophages of claim 11 or an engineered or differentiated cell thereof to a subject in need thereof, thereby treating the disease.

### Claim 19 (depends on 15)

A method of treating a disease, the method comprising administering the recovered cells of claim 15 or an engineered or differentiated cell thereof to a subject in need thereof, thereby treating the disease.

### Claim 20

A method of delivering a therapeutic agent, the method comprising administering an aptamer comprising a sequence having at least 75% sequence identity to SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO:3, wherein the aptamer is attached to a therapeutic drug or therapeutic cell;
wherein any nucleotide change in the sequence of the aptamer relative to SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO:3 occurs within one or more stem regions of SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO:3 and co-occurs with one or more compensatory nucleotide changes within the one or more stem regions of SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO:3 to maintain double-strandedness of the one or more stem regions of SEQ ID NO:1, SEQ ID NO: 2, or SEQ ID NO: 3.