# US 12,006,511 B2 · Recombinant PD-L1 peptides and methods of use

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- Number: US 12,006,511 B2
- Document ID: US12006511B2
- Status: Granted
- Kind: B2
- Application number: 17/046,287
- Filed: 2020-04-14
- Published: 2023-05-04
- Granted: 2024-06-11
- Earliest claimed: 2019-04-15
- Assignees: The Medical College of Wisconsin, Inc.; University of Virginia Patent Foundation
- Applicants: The Medical College of Wisconsin, Inc.; University of Virginia Patent Foundation
- Inventors: Kaothara Oluwakemi Adeyanju; Jeffrey A. Medin; Robert Blake Hill; Lawrence G. Lum
- Web page: https://patentgrep.com/patent/US12006511B2

## CPC

- C07K 14/70532: {B7 molecules, e.g. CD80, CD86}
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/22: Immunosuppressive or immunotolerising
- A61K 40/416: {Antigens related to auto-immune diseases; Preparations to induce self-tolerance}
- C12N 5/0636: {T lymphocytes}
- C12N 2501/2302: Interleukin-2 (IL-2)
- C12N 2501/24: Interferons [IFN]
- C12N 2501/48: Regulators of apoptosis
- C12N 2501/505: CD4; CD8
- C12N 2501/51: B7 molecules, e.g. CD80, CD86, CD28 (ligand), CD152 (ligand)
- C12N 2501/515: CD3, T-cell receptor complex
- C12N 2531/00: Microcarriers
- 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)
- 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)
- 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 disclosure provides soluble truncated mutant programmed death-ligand 1 (PD-L1) peptides. Polynucleotides and vectors encoding the soluble truncated mutant PD-L1 peptides are also provided. Further, methods of using the soluble truncated mutant peptides to promote differentiation of CD4+ effector T (Th1) cells into Foxp3+ regulatory T (Treg) cells are provided.

## Claims

### Claim 1

A soluble mutant programmed death-ligand 1(PD-L1)peptide consisting essentially of SEQ ID NO: 1, wherein the PD-L1 peptide comprises one or more mutations selected from the group consisting of D9E, D105E, Y106W, R108M, Y39I, S1001, V51I, and combinations thereof, and wherein the position of each mutation is numbered relative to SEQ ID NO: 1.

### Claim 2 (depends on 1)

The soluble mutant PD-L1 peptide of claim 1, wherein the PD-L1 peptide comprises two or more mutations selected from the group consisting of D9E, D105E, Y106W, R108M, Y39I, S100I, V51I, and combinations thereof.

### Claim 3 (depends on 1)

The soluble mutant PD-L1 peptide of claim 1, wherein the PD-L1 peptide comprises three or more mutations selected from the group consisting of D9E, D105E, Y106W, R108M, Y39I, S100I, and combinations thereof.

### Claim 4 (depends on 1)

The soluble mutant PD-L1 peptide of claim 1, wherein the PD-L1 peptide comprises four or more mutations selected from the group consisting of D9E, D105E, Y106W, R108M, Y39I, S100I, V51I, and combinations thereof.

### Claim 5 (depends on 1)

The soluble mutant PD-L1 peptide of claim 1, wherein the PD-L1 peptide comprises five or more mutations selected from the group consisting of D9E, D105E, Y106W, R108M, Y39I, S1001, V51I, and combinations thereof.

### Claim 6 (depends on 1)

The soluble mutant PD-L1 peptide of claim 1, wherein the PD-L1 peptide comprises six or more mutations selected from the group consisting of D9E, D105E, Y106W, R108M, Y39I, S1001, V51I, and combinations thereof.

### Claim 7 (depends on 1)

The soluble mutant PD-L1 peptide of claim 1, wherein the PD-L1 peptide comprises the following mutations: D9E, D105E, Y106W, R108M, Y39I, S1001, and V51I.

### Claim 8 (depends on 1)

The soluble mutant PD-L1 peptide of claim 1, wherein the peptide consists essentially of an amino acid sequence selected from:
(a)
(PDL1A; SEQ ID NO: 2)
AFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEMEDKNIIQ
FVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCM
ISYGGADWKRITVKVNAPY,
(b)
(PDL1C; SEQ ID NO: 3)
AFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVIWEMEDKNIIQ
FIHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCM
IIYGGADWKRITVKVNAPY,
and
(c)
(PDL1D; SEQ ID NO: 4)
AFTVTVPKELYVVEYGSNMTIECKFPVEKQLDLAALIVIWEMEDKNIIQ
FIHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCM
IIYGGAEWKMITVKVNAPY.

### Claim 9 (depends on 1)

The soluble mutant PD-L1 peptide of claim 1, wherein the peptide further comprises a tag or agent.

### Claim 10 (depends on 1)

A polynucleotide encoding the soluble mutant PD-L1 peptide of claim 1.

### Claim 11 (depends on 1)

A vector capable of expressing the soluble mutant PD-L1 peptide, the vector comprising a promoter operably connected to a polynucleotide encoding the soluble mutant PD-L1 peptide of claim 1.

### Claim 12 (depends on 11)

The vector of claim 11, further comprising heterologous backbone sequence.

### Claim 13 (depends on 11)

A host cell comprising the vector of claim 11.

### Claim 14 (depends on 1)

A soluble mutant PD-L1 peptide-bead complex, the complex comprising the soluble mutant PD-L1 peptide of claim 1 and a bead, wherein the peptide is covalently or non-covalently attached to the bead.

### Claim 15 (depends on 1, 14)

A method of generating regulatory Foxp3+T cells (Foxp3+Tregs), the method comprising contacting T cells cultured in vitro with the soluble mutant PD-L1 peptide of claim 1 or the soluble mutant PD-L1 peptide-bead complex of claim 14 in an amount effective to generate Treg cells, wherein the soluble mutant PD-L1 peptide comprises SEQ ID NO: 2 (PDL1A), SEQ ID NO: 3 (PDL1C), or SEQ ID NO: 4 (PDL1D).