# US 12,006,354 B2 · Antibody-IL2 engrafted proteins and methods of use in the treatment of cancer

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- Number: US 12,006,354 B2
- Document ID: US12006354B2
- Status: Granted
- Kind: B2
- Application number: 16/616,318
- Filed: 2018-05-22
- Published: 2020-08-27
- Granted: 2024-06-11
- Earliest claimed: 2017-05-24
- Assignees: Novartis AG
- Applicants: Novartis AG
- Inventors: Jonathan Deane; Yaiza Diaz-De-Durana; Michael DiDonato; Christophe Filippi; Glen Spraggon
- Web page: https://patentgrep.com/patent/US12006354B2

## CPC

- A61K 47/6813: {the drug being a peptidic cytokine, e.g. an interleukin or interferon}
- A61K 38/2013: {IL-2}
- A61K 39/3955: {against proteinaceous materials, e.g. enzymes, hormones, lymphokines}
- A61P 35/00: Antineoplastic agents
- C07K 16/46: Hybrid immunoglobulins (hybrids of an immunoglobulin with a peptide not being an immunoglobulin C07K19/00)
- A61K 2039/505: {comprising antibodies}
- C07K 14/55: IL-2
- C07K 16/11: Paramyxoviridae (F); Pneumoviridae (F), e.g. respiratory syncytial virus [RSV]
- C07K 2317/56: variable (Fv) region, i.e. VH and/or VL
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/74: Inducing cell proliferation
- C07K 2317/90: characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K 2317/94: Stability, e.g. half-life, pH, temperature or enzyme-resistance
- C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
- C07K 2319/31: fusions, other than Fc, for prolonged plasma life, e.g. albumin
- C12N 5/0636: {T lymphocytes}
- C12N 5/0682: {Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells (oocytes C12N5/0609)}
- 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 disclosure provides for IL2 engrafted into the CDR sequences of an antibody having preferred therapeutic profiles over molecules known and used in the clinic. In particular, the provided antibody cytokine engrafted protein compositions increase or maintain CD8+ T effector cells while reducing the activity of Treg cells. Additionally, provided compositions convey improved half-life, stability and producibility over recombinant human IL2 formulations such as Proleukin®.

## Claims

### Claim 1

An antibody cytokine engrafted protein comprising:
(a) an IgG class heavy chain comprising an IgG class heavy chain variable region (VH), comprising Complementarity Determining Regions (CDR) HCDR1 comprising SEQ ID NO: 7, SEQ ID NO: 10, SEQ ID NO: 13, SEQ ID NO: 16, SEQ ID NO: 39, SEQ ID NO: 42, SEQ ID NO: 45, or SEQ ID NO: 48, HCDR2 comprising SEQ ID NO: 8, SEQ ID NO: 11, SEQ ID NO: 14, SEQ ID NO: 17, SEQ ID NO: 40, SEQ ID NO: 46 or SEQ ID NO: 49, and HCDR3 comprising SEQ ID NO: 9, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 18, SEQ ID NO: 41, SEQ ID NO: 44, SEQ ID NO: 47 or SEQ ID NO: 50, and wherein the HCDRs individually comprise between 0 to 1 substitution; and
(b) an IgG class light chain comprising an IgG class light chain variable region (VL), comprising LCDR1 comprising SEQ ID NO: 23, SEQ ID NO: 26, SEQ ID NO: 29, SEQ ID NO: 32, SEQ ID NO: 55, SEQ ID NO: 58, SEQ ID NO: 61 or SEQ ID NO: 64, LCDR2 comprising SEQ ID NO: SEQ ID NO: 24, SEQ ID NO: 27, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 56, SEQ ID NO: 59, SEQ ID NO: 62 or SEQ ID NO: 65, and LCDR3 comprising SEQ ID NO: 25, SEQ ID NO: 28, SEQ ID NO: 31, SEQ ID NO: 34, SEQ ID NO: 57, SEQ ID NO: 60, SEQ ID NO: 63 or SEQ ID NO: 66, and wherein the LCDRs individually comprise between 0 to 1 substitution; and
(c) a mutated Interleukin 2 (IL2) molecule engrafted into HCDR1, wherein the HCDR1 comprises SEQ ID NO: 7, SEQ ID NO: 10, SEQ ID NO: 13, SEQ ID NO: 16, SEQ ID NO: 39, SEQ ID NO: 42, SEQ ID NO: 45, or SEQ ID NO: 48, and wherein the mutated IL2 molecule contains a mutation that reduces the affinity of the mutated IL2 molecule to a high affinity IL2 receptor.

### Claim 2

An antibody cytokine engrafted protein comprising:
(a) an IgG class heavy chain comprising an IgG class heavy chain variable region (VH), comprising Complementarity Determining Regions (CDR) HCDR1 comprising SEQ ID NO: 7, SEQ ID NO: 10, SEQ ID NO: 13, SEQ ID NO: 16, SEQ ID NO: 39, SEQ ID NO: 42, SEQ ID NO: 45, or SEQ ID NO: 48, HCDR2 comprising SEQ ID NO: 8, SEQ ID NO: 11, SEQ ID NO: 14, SEQ ID NO: 17, SEQ ID NO: 40, SEQ ID NO: 46 or SEQ ID NO: 49, and HCDR3 comprising SEQ ID NO: 9, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 18, SEQ ID NO: 41, SEQ ID NO: 44, SEQ ID NO: 47 or SEQ ID NO: 50, and wherein the HCDRs individually comprise between 0 to 1 substitution; and
(b) an IgG class light chain comprising an IgG class light chain variable region (VL), comprising LCDR1 comprising SEQ ID NO: 23, SEQ ID NO: 26, SEQ ID NO: 29, SEQ ID NO: 32, SEQ ID NO: 55, SEQ ID NO: 58, SEQ ID NO: 61 or SEQ ID NO: 64, LCDR2 comprising SEQ ID NO: SEQ ID NO: 24, SEQ ID NO: 27, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 56, SEQ ID NO: 59, SEQ ID NO: 62 or SEQ ID NO: 65, and LCDR3 comprising SEQ ID NO: 25, SEQ ID NO: 28, SEQ ID NO: 31, SEQ ID NO: 34, SEQ ID NO: 57, SEQ ID NO: 60, SEQ ID NO: 63 or SEQ ID NO: 66, and wherein the LCDRs individually comprise between 0 to 1 substitution, and
a mutated Interleukin 2 (IL2) molecule engrafted into HCDR1, wherein the HCDR1 comprises SEQ ID NO: 7, SEQ ID NO: 10, SEQ ID NO: 13, SEQ ID NO: 16, SEQ ID NO: 39, SEQ ID NO: 42, SEQ ID NO: 45, or SEQ ID NO: 48, wherein the antibody cytokine engrafted protein preferentially expands T effector cells over regulatory T cells.

### Claim 3

An antibody cytokine engrafted protein comprising:
(a) an IgG class heavy chain comprising an IgG class heavy chain variable region (VH), comprising Complementarity Determining Regions (CDR) HCDR1 comprising SEQ ID NO: 7, SEQ ID NO: 10, SEQ ID NO: 13, SEQ ID NO: 16, SEQ ID NO: 39, SEQ ID NO: 42, SEQ ID NO: 45, or SEQ ID NO: 48, HCDR2 comprising SEQ ID NO: 8, SEQ ID NO: 11, SEQ ID NO: 14, SEQ ID NO: 17, SEQ ID NO: 40, SEQ ID NO: 46 or SEQ ID NO: 49, and HCDR3 comprising SEQ ID NO: 9, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 18, SEQ ID NO: 41, SEQ ID NO: 44, SEQ ID NO: 47 or SEQ ID NO: 50, and wherein the HCDRs individually comprise between 0 to 1 substitution; and
(b) an IgG class light chain comprising an IgG class light chain variable region (VL), comprising LCDR1 comprising SEQ ID NO: 23, SEQ ID NO: 26, SEQ ID NO: 29, SEQ ID NO: 32, SEQ ID NO: 55, SEQ ID NO: 58, SEQ ID NO: 61 or SEQ ID NO: 64, LCDR2 comprising SEQ ID NO: SEQ ID NO: 24, SEQ ID NO: 27, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO: 56, SEQ ID NO: 59, SEQ ID NO: 62 or SEQ ID NO: 65, and LCDR3 comprising SEQ ID NO: 25, SEQ ID NO: 28, SEQ ID NO: 31, SEQ ID NO: 34, SEQ ID NO: 57, SEQ ID NO: 60, SEQ ID NO: 63 or SEQ ID NO: 66, and wherein the LCDRs individually comprise between 0 to 1 substitution; and
(c) a mutated Interleukin 2 (IL2) molecule engrafted into HCDR1, wherein the HCDR1 comprises SEQ ID NO: 7, SEQ ID NO: 10, SEQ ID NO: 13, SEQ ID NO: 16, SEQ ID NO: 39, SEQ ID NO: 42, SEQ ID NO: 45, or SEQ ID NO: 48;
wherein the antibody cytokine engrafted protein stimulates one or more of CD8+ T effector cell proliferation, natural killer (NK) cell proliferation or combination thereof, greater than recombinant IL2 or aldesleukin.

### Claim 4 (depends on 1)

The antibody cytokine engrafted protein of claim 1, wherein the VH comprises SEQ ID NO: 19 or SEQ ID NO: 51.

### Claim 5 (depends on 1)

The antibody cytokine engrafted protein of claim 1, wherein the VL comprises SEQ ID NO: 35, or SEQ ID NO: 67.

### Claim 6 (depends on 1)

The antibody cytokine engrafted protein of claim 1, wherein the IgG class heavy chain is selected from IgG1, IgG2, and IgG4.

### Claim 7 (depends on 1)

The antibody cytokine engrafted protein of claim 1, wherein the IgG class heavy chain comprises SEQ ID NO: 21 or SEQ ID NO: 53.

### Claim 8 (depends on 1)

The antibody cytokine engrafted protein of claim 1, wherein the IgG class light chain comprises SEQ ID NO:69 and the IgG class heavy chain comprises SEQ ID NO:53.

### Claim 9 (depends on 1)

The antibody cytokine engrafted protein of claim 1, wherein the IgG class light chain comprises SEQ ID NO:37 and the IgG class heavy chain comprises SEQ ID NO:21.

### Claim 10 (depends on 1)

The antibody cytokine engrafted protein of claim 1, wherein the IgG class light chain comprises SEQ ID NO:69 and the IgG class heavy chain comprises SEQ ID NO:21.

### Claim 11 (depends on 1)

The antibody cytokine engrafted protein of claim 1, wherein the IgG class light chain comprises SEQ ID NO:37 and the IgG class heavy chain comprises SEQ ID NO:53.

### Claim 12 (depends on 1)

The antibody cytokine engrafted protein of claim 1, wherein the IgG class light chain comprises 95% or more sequence identity with SEQ ID NO:37 and the IgG class heavy chain comprises 95% or more sequence identity with SEQ ID NO:21.

### Claim 13 (depends on 1)

A method of treating cancer in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of the antibody cytokine engrafted protein of claim 1.

### Claim 14 (depends on 13)

The method of claim 13, wherein the cancer is selected from the group consisting of melanoma, lung cancer, colorectal cancer, prostate cancer, breast cancer, and lymphoma.

### Claim 15 (depends on 13)

The method of claim 13, wherein the antibody cytokine engrafted protein is administered in combination with a therapeutic agent.

### Claim 16 (depends on 15)

The method of claim 15, wherein the therapeutic agent is a second antibody cytokine engrafted protein.

### Claim 17 (depends on 15)

The method of claim 15, wherein the therapeutic agent is an immune checkpoint inhibitor.

### Claim 18 (depends on 17)

The method of claim 17, wherein the immune checkpoint is selected from the group consisting of: PD-1, PD-L1, PD-L2, TIM3, CTLA-4, LAG-3, CEACAM-1, CEACAM-5, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and TGFR.

### Claim 19 (depends on 15)

The method of claim 15, wherein the therapeutic agent is a tyrosine kinase inhibitor.

### Claim 20 (depends on 1)

A pharmaceutical composition comprising the antibody cytokine engrafted protein of claim 1 and a pharmaceutically acceptable carrier.

### Claim 21 (depends on 20)

A method of treating cancer in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of the pharmaceutical composition of claim 20.

### Claim 22 (depends on 21)

The method of claim 21, wherein the cancer is selected from the group consisting of melanoma, lung cancer, colorectal cancer, prostate cancer, breast cancer, and lymphoma.

### Claim 23 (depends on 21)

The method of claim 21, wherein the pharmaceutical composition is administered in combination with a therapeutic agent.

### Claim 24 (depends on 23)

The method of claim 23, wherein the therapeutic agent is a second antibody cytokine engrafted protein.

### Claim 25 (depends on 23)

The method of claim 23, wherein the therapeutic agent is an immune checkpoint inhibitor.

### Claim 26 (depends on 25)

The method of claim 25, wherein the immune checkpoint is selected from the group consisting of: PD-1, PD-L1, PD-L2, TIM3, CTLA-4, LAG-3, CEACAM-1,CEACAM-5, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and TGFR.

### Claim 27 (depends on 23)

The method of claim 23, wherein the therapeutic agent is a tyrosine kinase inhibitor.