# US 8,771,687 B2 · Methods for treating a tumor using an antibody that specifically binds GRP94

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- Number: US 8,771,687 B2
- Document ID: US8771687B2
- Status: Granted
- Kind: B2
- Application number: 13/309,490
- Filed: 2011-12-01
- Published: 2012-06-14
- Granted: 2014-07-08
- Earliest claimed: 2010-12-02
- Assignees: University of Pittsburgh—Of the Commonwealth System of Higher Education
- Applicants: Soldano Ferrone; Xinhui Wang; Elvira Favoino; Ling Yu; Yangyang Wang
- Inventors: Soldano Ferrone; Xinhui Wang; Elvira Favoino; Ling Yu; Yangyang Wang
- Web page: https://patentgrep.com/patent/US8771687B2

## CPC

- C07K 16/18: against material from animals or humans
- A61K 31/437: the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
- A61K 39/3955: {against proteinaceous materials, e.g. enzymes, hormones, lymphokines}
- A61K 39/39558: {against tumor tissues, cells, antigens}
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61P 35/00: Antineoplastic agents
- A61P 35/04: specific for metastasis
- C07K 16/3053: {Skin, nerves, brain}
- A61K 2039/505: {comprising antibodies}
- C07K 2317/21: from primates, e.g. man
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Combinations of agents that have a synergistic effect for the treatment of a tumor are disclosed herein. These combinations of agents can be used to treat tumors, wherein the cells of the cancer express a mutated BRAF. Methods are disclosed for treating a subject diagnosed with a tumor that expresses a mutated BRAF. The methods include administering to the subject (1) a therapeutically effective amount of an antibody or antigen binding fragment thereof that specifically binds glucose regulated protein (GRP) 94; and (2) a therapeutically effective amount of a BRAF inhibitor. In some embodiments, the tumor is melanoma. In some embodiments the method includes selecting a subject with primary or secondary resistance to a BRAF inhibitor. In further embodiments, treating the tumor comprises decreasing the metastasis of the tumor. In additional embodiments, the BRAF inhibitor comprises PLX4032 or PLX4720.

## Claims

### Claim 1

A method of treating a subject diagnosed with a tumor that expresses a BRAF mutation, comprising
administering to the subject (1) a therapeutically effective amount of a monoclonal antibody or antigen binding fragment thereof that specifically binds glucose regulated protein (GRP) 94; and (2) a therapeutically effective amount of a BRAF inhibitor,
thereby treating the tumor in the subject, wherein the tumor is melanoma.

### Claim 2 (depends on 1)

The method of claim 1, wherein the subject has primary or secondary resistance to the BRAF inhibitor.

### Claim 3 (depends on 1)

The method of claim 1, wherein the BRAF inhibitor is PLX4032 or PLX4720.

### Claim 4 (depends on 1)

The method of claim 1, comprising administering to the subject a therapeutically effective amount of the antigen binding fragment of the monoclonal antibody that specifically binds GRP94.

### Claim 5 (depends on 1)

The method of claim 1, wherein the monoclonal antibody or antigen binding fragment comprises a heavy chain variable domain, and wherein the heavy chain variable domain of the monoclonal antibody comprises the amino acid sequence set forth as amino acids 26-33 of SEQ ID NO: 3, amino acids 51-58 of SEQ ID NO: 3, and amino acids 97-103 of SEQ ID NO: 3.

### Claim 6 (depends on 1)

The method of claim 1, wherein the monoclonal antibody or antigen binding fragment comprises a light chain variable domain, wherein the light chain variable domain of the antibody comprises the amino acid sequence set forth as amino acids 27-32 of SEQ ID NO: 4, amino acids 50-52 of SEQ ID NO: 4, and amino acids 89-97 of SEQ ID NO: 4.

### Claim 7 (depends on 1)

The method of claim 1, wherein treating the tumor comprises decreasing the metastasis of the tumor.

### Claim 8 (depends on 1)

The method of claim 1, wherein the subject is human.

### Claim 9 (depends on 1)

The method of claim 1, wherein the melanoma is superficial spreading melanoma, nodular melanoma, acral lentiginous melanoma, lentiginous malignant melanoma, or mucosal lentiginous melanoma.

### Claim 10

The method of claim, wherein the melanoma is cutaneous or extracutaneous melanoma.

### Claim 11 (depends on 1)

The method of claim 1, wherein the melanoma is intraocular or a clear-cell sarcoma of the soft tissues.

### Claim 12 (depends on 1)

The method of claim 1, further comprising administering one or more additional chemotherapeutic agents.

### Claim 13 (depends on 1)

The method of claim 1, wherein the therapeutically effective amount of the BRAF inhibitor and the therapeutically effective amount of the antibody or antigen binding fragment thereof that specifically binds glucose regulated protein (GRP) 94 are administered simultaneously.

### Claim 14 (depends on 1)

The method of claim 1, wherein cells in the tumor comprise a BRAF mutation.

### Claim 15 (depends on 14)

The method of claim 14, wherein the BRAF mutation is a V600E mutation.

### Claim 16 (depends on 14)

The method of claim 14, wherein the BRAF mutation is R462I, I463S, G464E, G464V, G466A, G466E, G466V, G469A, G469E, N581S, E585K, D594V, F595L, G596R, L597V, T599I, V600D, V600E, V600K, V600R, K601E or A728V.

### Claim 17 (depends on 14)

The method of claim 14, further comprising
detecting a V600E BRAF mutation in a sample from the subject, wherein the sample comprises cells from the tumor.

### Claim 18 (depends on 1)

The method of claim 1, wherein:
the BRAF inhibitor is PLX4032 or PLX4720; and
the monoclonal antibody, or antigen binding fragment thereof comprises a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain of the monoclonal antibody comprises the amino acid sequence set forth as amino acids 26-33 of SEQ ID NO: 3, amino acids 51-58 of SEQ ID NO: 3, and amino acids 97-103 of SEQ ID NO: 3 and the light chain variable domain of the antibody comprises the amino acid sequence set forth as amino acids 27-32 of SEQ ID NO: 4, amino acids 50-52 of SEQ ID NO: 4, and amino acids 89-97 of SEQ ID NO: 4.

### Claim 19

A pharmaceutical composition comprising (1) a therapeutically effective amount of a monoclonal antibody or antigen binding fragment thereof that specifically binds glucose regulated protein (GRP) 94; and (2) a therapeutically effective amount of a BRAF inhibitor.

### Claim 20 (depends on 19)

The pharmaceutical composition of claim 19, wherein the BRAF inhibitor is PLX4032 or PLX4720.

### Claim 21 (depends on 20)

The pharmaceutical composition of claim 20, wherein
the BRAF inhibitor is PLX4032; and
the monoclonal antibody, or antigen binding fragment thereof comprises a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain of the monoclonal antibody comprises the amino acid sequence set forth as amino acids 26-33 of SEQ ID NO: 3, amino acids 51-58 of SEQ ID NO: 3, and amino acids 97-103 of SEQ ID NO: 3 and the light chain variable domain of the antibody comprises the amino acid sequence set forth as amino acids 27-32 of SEQ ID NO: 4, amino acids 50-52 of SEQ ID NO: 4, and amino acids 89-97 of SEQ ID NO: 4.

### Claim 22 (depends on 19)

The pharmaceutical composition of claim 19, formulated for systemic administration.