# US 10,501,538 B2 · Anti-LILRB antibodies and their use in detecting and treating cancer

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

- Number: US 10,501,538 B2
- Document ID: US10501538B2
- Status: Granted
- Kind: B2
- Application number: 15/696,972
- Filed: 2017-09-06
- Published: 2018-03-29
- Granted: 2019-12-10
- Earliest claimed: 2015-03-06
- Assignees: Board of Regents, The University of Texas System
- Applicants: The Board of Regents of the University of Texas System
- Inventors: Chengcheng Zhang; Mi Deng; Wei Xiong; Zhiqiang An; Ningyan Zhang; Xun Gui; Junke Zheng
- Web page: https://patentgrep.com/patent/US10501538B2

## CPC

- C07K 16/2803: {against the immunoglobulin superfamily}
- A61K 39/3955: {against proteinaceous materials, e.g. enzymes, hormones, lymphokines}
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61P 35/02: specific for leukemia
- C07K 16/3061: {Blood cells}
- C12N 15/1138: {against receptors or cell surface proteins}
- G01N 33/57505: of the blood, e.g. leukaemia
- G01N 33/5759: {involving compounds localised on the membrane of tumour or cancer cells}
- A61K 2039/505: {comprising antibodies}
- C07K 2317/14: Specific host cells or culture conditions, e.g. components, pH or temperature
- C07K 2317/21: from primates, e.g. man
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C07K 2317/33: Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
- C07K 2317/54: F(ab')2
- C07K 2317/55: Fab or Fab'
- C07K 2317/56: variable (Fv) region, i.e. VH and/or VL
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/531: Stem-loop; Hairpin
- G01N 2333/70503: Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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

The present disclosure is directed to antibodies binding to LILRBs and methods of detecting and treating cancer therewith.

## Claims

### Claim 1

An isolated monoclonal antibody or an antigen-binding fragment that binds to LILRB4 thereof comprising:
(a) a heavy chain variable region comprising the complementary determining regions (CDRs) of SEQ ID NOS: 73, 74 and 75 in that order; and
a light chain variable region comprising the CDR of SEQ ID NO: 76, TAS and SEQ ID NO: 77 in that order;
(b) a heavy chain variable region comprising the CDRs of SEQ ID NOS: 83, 84 and 85 in that order; and a light chain variable region comprising the CDRs of SEQ ID NO: 86, RAS and SEQ ID NO: 87 in that order;
(c) a heavy chain variable region comprising the CDRs of SEQ ID NOS: 98, 99 and 100 in that order; and a light chain variable region comprising the CDRs of SEQ ID NO: 101, EAS and SEQ ID NO: 102 in that order; or
(d) a heavy chain variable region comprising the CDRs of SEQ ID NOS: 166, 167 and 168 in that order; and a light chain variable region comprising the CDRs of SEQ ID NOS: 169, 170 and 171 in that order.

### Claim 2 (depends on 1)

The isolated monoclonal antibody or an antigen-binding fragment thereof of claim 1, wherein the antigen-binding fragment is a recombinant ScFv (single chain fragment variable) antibody, Fab fragment, F(ab')2 fragment, or Fv fragment.

### Claim 3 (depends on 1)

The isolated monoclonal antibody or an antigen binding fragment thereof of claim 1, wherein the isolated monoclonal antibody is a chimeric, humanized, or human antibody.

### Claim 4 (depends on 1)

A pharmaceutical composition comprising the isolated monoclonal antibody or an antigen-binding fragment thereof of claim 1, and a pharmaceutically acceptable carrier.

### Claim 5 (depends on 1)

An isolated nucleic acid that encodes the isolated monoclonal antibody of claim 1.

### Claim 6 (depends on 5)

A vector comprising the isolated nucleic acid of claim 5.

### Claim 7 (depends on 6)

A host cell comprising the vector of claim 6.

### Claim 8 (depends on 7)

The host cell of claim 7, wherein the host cell is a mammalian cell.

### Claim 9 (depends on 7)

The host cell of claim 7, wherein the host cell is a CHO cell.

### Claim 10 (depends on 1)

A hybridoma or engineered cell encoding or producing the isolated monoclonal antibody of claim 1.

### Claim 11 (depends on 7)

A process of producing an antibody, comprising culturing the host cell of claim 7 under conditions suitable for expressing the antibody, and recovering the antibody.

### Claim 12 (depends on 1)

A method of treating or ameliorating the effect of a cancer in a subject, the method comprising administering to said subject a therapeutically effective amount of the antibody or an antigen-binding fragment thereof of claim 1.

### Claim 13 (depends on 12)

The method of claim 12, wherein the cancer is acute myeloid leukemia.

### Claim 14 (depends on 12)

The method of claim 12, wherein the antibody or an antigen-binding fragment thereof is administered intravenously, intra-arterially, intra-tumorally, or subcutaneously.

### Claim 15 (depends on 12)

The method of claim 12, further comprising administering to the subject one or more drugs selected from the group consisting of an anthracycline topoisomerase inhibitor, a daunorubicin, a nucleoside metabolic inhibitor, a cytarabine, a combination of daunorubicin and cytarabine, a daunorubicin and cytarabine liposome for injection, Vyxeos, an all-trans-retinoic acid (ATRA), an arsenic, an arsenic trioxide, a histamine dihydrochloride, Ceplene, an interleukin-2, Proleukin, a gemtuzumab ozogamicin, Mylotarg, a clofarabine, a farnesyl transferase inhibitor, a decitabine, an IDH1 inhibitor, an IDH2 inhibitor, an enasidenib, Idhifa, an IDO inhibitor, an epacadostat, a platinum complex derivative, oxaliplatin, a kinase inhibitor, a tyrosine kinase inhibitor, a PI3 kinase inhibitor, a BTK inhibitor, ibrutinib, a PD-1 antibody, a PD-L1 antibody, a CTLA-4 antibody, a LAG3 antibody, an ICOS antibody, a TIGIT antibody, a TIM3 antibody, an antibody binding to a tumor antigen, an antibody binding to a T-cell surface marker, an antibody binding to a myeloid cell or NK cell surface marker, an alkylating agent, a nitrosourea agent, an antimetabolite, an antitumor antibiotic, an alkaloid derived from a plant, a topoisomerase inhibitor, a hormone therapy medicine, a hormone antagonist, an aromatase inhibitor, and a P-glycoprotein inhibitor.

### Claim 16 (depends on 1)

A method of detecting a cancer cell or cancer stem cell in a sample or subject comprising:
(a) contacting a subject or a sample from the subject with the antibody or an antigen-binding fragment thereof of claim 1; and
(b) detecting binding of said antibody to a cancer cell or cancer stem cell in said subject or sample.

### Claim 17 (depends on 16)

The method of claim 16, wherein the sample is a body fluid or biopsy.

### Claim 18 (depends on 16)

The method of claim 16, wherein the sample is blood, sputum, tears, saliva, mucous, serum, urine or feces.

### Claim 19 (depends on 16)

The method of claim 16, wherein detection comprises immunohistochemistry, ELISA, RIA or Western blot.

### Claim 20 (depends on 16)

The method of claim 16, further comprising performing steps (a) and (b) a second time and determining a change in detection levels as compared to the first time.

### Claim 21 (depends on 1)

The isolated monoclonal antibody or an antigen-binding fragment thereof of claim 1, further characterized as comprising:
(a) a heavy chain variable region having an amino acid sequence at least about 90% identical to SEQ ID NO: 13; and
a light chain variable region having an amino acid sequence at least about 90% identical to SEQ ID NO: 14;
(b) a heavy chain variable region having an amino acid sequence at least about 90% identical to SEQ ID NO: 17; and a light chain variable region having an amino acid sequence at least about 90% identical to SEQ ID NO: 18;
(c) a heavy chain variable region having an amino acid sequence at least about 90% identical to SEQ ID NO: 23; and a light chain variable region having an amino acid sequence at least about 90% identical to SEQ ID NO: 24; or
(d) a heavy chain variable region having an amino acid sequence at least about 90% identical to SEQ ID NO: 222; and a light chain variable region having an amino acid sequence at least about 90% identical to SEQ ID NO: 223.

### Claim 22 (depends on 21)

The isolated monoclonal antibody or an antigen-binding fragment thereof of claim 21, wherein
(a) the heavy chain variable region has an amino acid sequence of SEQ ID NO: 13, and wherein the light chain variable region has an amino acid sequence of SEQ ID NO: 14 ;
(b) the heavy chain variable region has an amino acid sequence of SEQ ID NO: 17, and wherein the light chain variable region has an amino acid sequence of SEQ ID NO: 18;
(c) the heavy chain variable region has an amino acid sequence of SEQ ID NO: 23, and wherein the light chain variable region has an amino acid sequence of SEQ ID NO: 24; or
(d) the heavy chain variable region has an amino acid sequence of SEQ ID NO: 222, and wherein the light chain variable region has an amino acid sequence of SEQ ID NO: 223.

### Claim 23 (depends on 21)

The isolated monoclonal antibody or an antigen-binding fragment thereof of claim 21, wherein the antigen-binding fragment is a recombinant ScFv antibody, Fab fragment, F(ab′)2 fragment, or Fv fragment.

### Claim 24 (depends on 21)

The isolated monoclonal antibody or an antigen binding fragment thereof of claim 21, wherein the isolated monoclonal antibody is a chimeric, humanized, or human antibody.