# US 11,299,708 B2 · Methods for selective expansion of γδ T-cell populations and compositions thereof

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- Number: US 11,299,708 B2
- Document ID: US11299708B2
- Status: Granted
- Kind: B2
- Application number: 15/563,830
- Filed: 2017-05-12
- Published: 2019-04-25
- Granted: 2022-04-12
- Earliest claimed: 2016-05-12
- Assignees: Adicet Bio, Inc.
- Applicants: Adicet Bio, Inc.
- Inventors: Aya Jakobovits; Orit Foord; Andy An-deh Lin; Marianne Theresa Santaguida; Radhika Chetan Desai; Yifeng Frank Jing; Daulet Kadyl Satpayev; Yan Li
- Web page: https://patentgrep.com/patent/US11299708B2

## CPC

- A61K 35/17: Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00)
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/24: Antigen-presenting cells [APC]
- A61K 40/31: Chimeric antigen receptors [CAR]
- A61K 40/32: T-cell receptors [TCR]
- A61K 40/42: Cancer antigens
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 35/00: Antineoplastic agents
- C07K 16/2809: {against the T-cell receptor (TcR)-CD3 complex}
- C12N 5/0636: {T lymphocytes}
- A61K 2239/31: characterized by the route of administration
- A61K 2239/38: characterised by the dose, timing or administration schedule
- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 2317/74: Inducing cell proliferation
- C07K 2317/75: Agonist effect on antigen
- C07K 2319/03: containing a transmembrane segment
- 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
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention relates to methods for the selective expansion of γδ T-cell population(s), compositions and admixtures thereof and methods for using the same as a therapeutic. Non-engineered and engineered, enriched γδ T-cell populations of the disclosure are useful in the treatment of various cancers, infectious diseases, and immune disorders.

## Claims

### Claim 1

An ex vivo method for producing an enriched δ1 γδ T-cell population from an isolated mixed cell population without prior selection of γδ T-cells and/or depletion of non-γδ T-cells, and under culture conditions that do not comprise IL-4 and wherein said cell population has not been exposed to exogenous IL-4 prior to expansion, said method comprising directly contacting the mixed cell population with at least one antibody that selectively expands δ1 T-cells without significant expansion of δ2 T-cells by binding to an epitope specific to a δ1 variable region of a δ1 TCR to provide clinically-relevant levels of an enriched δ1 γδ T cell population, wherein the at least one antibody comprises a heavy chain variable region/light chain variable region (HCVR/LCVR) sequence pair selected from the group consisting of SEQ ID NOs: 153/180, 154/181, 155/182, 156/183, 157/184, 158/185, 159/186, 160/187, 161/188, 162/189, 163/190, 164/191, 165/192, 166/193, 167/194, 168/195, 169/196, 170/197, 171/198, 172/199, 173/200, 174/201, 175/202, 176/203, 177/204, 178/205, and 179/206.

### Claim 2 (depends on 1)

The method of claim 1, wherein the percentage of δ1 T cells in the enriched γδ T cell population is greater than 60% δ1 T-cells of total viable cells.

### Claim 3

The method of any one of the preceding claims, wherein said method does not comprise expanding the enriched δ1 γδ T-cell population by an antigen presenting cell or an aminophosphate.

### Claim 4 (depends on 1)

The method of claim 1, wherein said at least one antibody that selectively expands δ1 T-cells is immobilized on a surface.

### Claim 5 (depends on 1)

The method of claim 1, wherein said isolated mixed cell population is selected from a peripheral blood sample, a cord blood sample or a tumor.

### Claim 6 (depends on 1)

The method of claim 1, wherein said enriched δ1 γδ T-cell population comprise polyclonal TCR diversity.

### Claim 7 (depends on 1)

The method of claim 1, wherein the enriched δ1 γδ T-cell population is further formulated for administration to a subject.

### Claim 8 (depends on 1)

The method of claim 1, wherein the enriched δ1 γδ T-cell population includes a therapeutically effective amount of γδ T-cells.

### Claim 9 (depends on 1)

The method of claim 1, wherein said δ1 γδ T-cell population is further engineered to stably express one or more tumor recognition moieties.

### Claim 10 (depends on 1)

The method of claim 1, further comprising:
directly contacting at least a portion of the enriched δ1 γδ T-cell population with antigen presenting cells (APCs) in a second γδ T-cell expansion, thereby producing a second enriched γδ T-cell population,
wherein the second enriched γδ T-cell population comprises a clinically relevant number of γδ T-cells.

### Claim 11 (depends on 1)

The method of claim 1, further comprising:
directly contacting at least a portion of the enriched δ1 γδ T-cell population with one or more agents that (a) expand γδ T-cells, and/or (b) deplete αβ T-cells.

### Claim 12 (depends on 1)

The method of claim 1, wherein an antibody that selectively expands δ1 T-cells is an antibody that specifically recognizes the δ1 chain variable region and i) the CDR3 loop created by the restricted juxtaposition of V(D)J1 segments, or ii) the CDR3 loop created by the juxtaposition of V(D)J1 and V(D)J2 but not V(D)J3 segments, or iii) the CDR3 loop created by the juxtaposition of V(D)J1, V(D)J2 and V(D)J3 segments.

### Claim 13 (depends on 1)

The method of claim 1, wherein the antibody binds to an epitope comprising residues Arg71, Asp72 and Lys120 of the δ1 variable region.

### Claim 14 (depends on 1)

The method of claim 1, wherein the antibody has reduced binding to a mutant 61 TCR polypeptide comprising a mutation at K120 at the delta J1 and delta J2.

### Claim 15 (depends on 1)

The method of claim 1, wherein the epitope is selected from a δ1 TCR Bin 1 δ1 epitope, Bin 1b δ1 epitope, Bin 2 δ1 epitope, Bin 2b δ1 epitope, Bin 2c δ1 epitope, Bin 3 δ1 epitope, Bin 4 δ1 epitope, Bin 5 δ1 epitope, Bin 6 δ1 epitope, Bin 7 δ1 epitope, Bin 8 δ1 epitope, or Bin 9 δ1 epitope of a human δ1 TCR.