# US 2008/0188401 A1 · Compounds

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

- Number: US 2008/0188401 A1
- Document ID: US20080188401A1
- Status: Published application
- Kind: A1
- Application number: 11/948,659
- Filed: 2007-11-30
- Published: 2008-08-07
- Earliest claimed: 2006-11-30
- Assignees: MedImmune, LLC
- Inventors: Simon Charles Cruwys; Steven Godfrey Lane; Philip Mallinder
- Web page: https://patentgrep.com/patent/US20080188401A1

## CPC

- C07K 16/248: {IL-6}
- A61P 1/00: Drugs for disorders of the alimentary tract or the digestive system
- A61P 1/04: for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 3/04: Anorexiants; Antiobesity agents
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 5/14: of the thyroid hormones, e.g. T3, T4
- A61P 7/06: Antianaemics
- A61P 7/08: Plasma substitutes; Perfusion solutions; Dialytics or haemodialytics; Drugs for electrolytic or acid-base disorders, e.g. hypovolemic shock (artificial tears A61P27/04)
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 11/06: Antiasthmatics
- A61P 13/12: of the kidneys
- A61P 15/00: Drugs for genital or sexual disorders (for disorders of sex hormones A61P5/24); Contraceptives
- A61P 17/00: Drugs for dermatological disorders
- A61P 17/04: Antipruritics
- A61P 17/06: Antipsoriatics
- A61P 17/14: for baldness or alopecia
- A61P 19/00: Drugs for skeletal disorders
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 19/06: Antigout agents, e.g. antihyperuricemic or uricosuric agents
- A61P 21/00: Drugs for disorders of the muscular or neuromuscular system
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 27/00: Drugs for disorders of the senses
- 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 31/04: Antibacterial agents
- A61P 31/10: Antimycotics
- A61P 31/12: Antivirals
- A61P 31/16: for influenza or rhinoviruses
- A61P 31/18: for HIV
- A61P 33/00: Antiparasitic agents
- A61P 33/06: Antimalarials
- A61P 35/00: Antineoplastic agents
- A61P 35/02: specific for leukemia
- A61P 37/00: Drugs for immunological or allergic disorders
- A61P 37/02: Immunomodulators
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- G01N 33/6857: {Antibody fragments}
- A61K 2039/505: {comprising antibodies}
- C07K 2317/21: from primates, e.g. man
- C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
- C07K 2317/51: Complete heavy chain or Fd fragment, i.e. VH + CH1
- C07K 2317/515: Complete light chain, i.e. VL + CL
- C07K 2317/52: Constant or Fc region; Isotype
- C07K 2317/56: variable (Fv) region, i.e. VH and/or VL
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/567: Framework region [FR]
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- G01N 2333/5412: IL-6
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- 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)
- 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)

## Abstract

Binding members, e.g. human antibody molecules, which bind interleukin-6 (IL-6) and neutralise its biological effects. Use of binding members for IL-6 in medical treatment e.g. for treating inflammatory diseases and tumours associated with IL-6.

## Claims

### Claim 1

An isolated binding member for human IL-6, which binds human IL-6 with a KD of not more than 30 pM as defined by surface plasmon resonance.

### Claim 2 (depends on 1)

The binding member according to claim 1, wherein the KD is not more than 10 pM.

### Claim 3

An isolated binding member for human IL-6, which binds human IL-6 at residue Phe102 and/or Ser204, the residue numbering being defined according to the full length human IL-6 amino acid sequence SEQ ID NO: 161.

### Claim 4 (depends on 3)

The binding member according to claim 3, wherein the binding member binds human IL-6 at residue Arg207.

### Claim 5

An isolated binding member for human IL-6, wherein the binding member has an IC50 of less than 1 nM in an assay of inhibition of VEGF release from human synovial fibroblasts stimulated with 0.6 pM human IL-1β and 2.4 nM soluble human IL-6Rα.

### Claim 6 (depends on 1)

The binding member according to claim 1, wherein the binding member binds cynomolgus IL-6 and wherein cynomolgus IL-6 has an IC50 less than 10-fold different from the IC50 of unlabelled human IL-6 in a time resolved fluorescence assay for inhibition of labelled human IL-6 binding to the binding member immobilised on a support, wherein the labelled human IL-6 is at a final concentration equal to twice the Kd of its interaction with the binding member.

### Claim 7 (depends on 1)

The binding member according to claim 1, which has an IC50 of less than 100 pM in an assay for inhibition of proliferation of TF-1 cells in response to 20 pM human IL-6.

### Claim 8

An isolated binding member for human IL-6, comprising a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the set of CDRs has 22 or fewer amino acid alterations from a set of CDRs in which:
HCDR1 has amino acid sequence SEQ ID NO: 3;
HCDR2 has amino acid sequence SEQ ID NO: 4;
HCDR3 has amino acid sequence SEQ ID NO: 115;
LCDR1 has amino acid sequence SEQ ID NO: 8;
LCDR2 has amino acid sequence SEQ ID NO: 9; and
LCDR3 has amino acid sequence SEQ ID NO: 120, and wherein
the binding member has an IC50 of less than 100 pM in an assay for inhibition of proliferation of TF-1 cells in response to 20 pM human IL-6.

### Claim 9 (depends on 1)

The isolated binding member for human IL-6 according to claim 1, comprising a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the set of CDRs has 22 or fewer amino acid alterations from a set of CDRs in which:
HCDR1 has amino acid sequence SEQ ID NO: 3;
HCDR2 has amino acid sequence SEQ ID NO: 4;
HCDR3 has amino acid sequence SEQ ID NO: 115;
LCDR1 has amino acid sequence SEQ ID NO: 8;
LCDR2 has amino acid sequence SEQ ID NO: 9; and
LCDR3 has amino acid sequence SEQ ID NO: 120.

### Claim 10 (depends on 8)

The binding member according to claim 8, comprising a set of CDRs having 20 or fewer substitutions from a set of CDRs in which:
HCDR1 has amino acid sequence SEQ ID NO: 3;
HCDR2 has amino acid sequence SEQ ID NO: 4;
HCDR3 has amino acid sequence SEQ ID NO: 115;
LCDR1 has amino acid sequence SEQ ID NO: 8;
LCDR2 has amino acid sequence SEQ ID NO: 9; and
LCDR3 has amino acid sequence SEQ ID NO: 120;
and wherein the binding member optionally has an insertion of one residue to increase the length of the HCDR3 relative to SEQ ID NO: 115, and/or has an insertion of one residue to increase the length of the LCDR3 relative to SEQ ID NO: 120.

### Claim 11 (depends on 8)

The binding member according to claim 8, comprising an HCDR1 wherein Kabat residue 35 is Ile, Thr or Val.

### Claim 12 (depends on 11)

The binding member according to claim 11, wherein HCDR1 is SEQ ID NO: 3.

### Claim 13 (depends on 8)

The binding member according to claim 8, comprising an HCDR2 wherein Kabat residue 64 is Lys or Arg.

### Claim 14 (depends on 13)

The binding member according to claim 13, wherein HCDR2 has amino acid sequence SEQ ID NO: 4.

### Claim 15 (depends on 8)

The binding member according to claim 8, wherein:
Kabat residue 95 in HCDR3 is Trp and/or wherein Kabat residue 101 in HCDR3 is Asp.

### Claim 16 (depends on 8)

The binding member according to claim 8, comprising an HCDR3 wherein:
Kabat residue 96 is Ala or Glu;
Kabat residue 97 is Asp, Glu or Asn;
Kabat residue 98 is Asp, Gly, Glu or His;
Kabat residue 99 is His, Gly or Thr;
Kabat residue 100 is Pro, Tyr, Asn, Arg, Trp or Ala;
Kabat residue 100A is Pro, Tyr, Ala, Arg, Thr, Gly, Asn, Pro or Ser;
Kabat residue 100B is Trp, Tyr, His, Gln, Pro or Thr;
Kabat residue 100C is Ile, Ala, Val, His, Tyr or Leu; and
Kabat residue 102 is Leu, Val, His, Met or Ile.

### Claim 17 (depends on 16)

The binding member according to claim 16, wherein HCDR3 has amino acid sequence SEQ ID NO: 115.

### Claim 18 (depends on 8)

The binding member according to claim 8, wherein Kabat residue 34 in LCDR1 is Ala or Thr.

### Claim 19 (depends on 18)

The binding member according to claim 18, wherein LCDR1 is SEQ ID NO: 8.

### Claim 20 (depends on 8)

The binding member according to claim 8, wherein LCDR2 is SEQ ID NO: 9.

### Claim 21 (depends on 8)

The binding member according to claim 8, comprising an LCDR3 wherein:
Kabat residue 89 is Gln, Met or Ala;
Kabat residue 90 is Gln, Asn, Ser or Ala;
Kabat residue 91 is Ser, Asn, Gly, Ala or His;
Kabat residue 92 is Trp, Tyr, Ser, Lys or Phe;
Kabat residue 93 is Leu, Ser, Lys, Arg or Ala;
Kabat residue 94 is Gly, Thr, Ala or Pro;
Kabat residue 96 is Gly or Trp; and
Kabat residue 97 is Ser or Thr.

### Claim 22 (depends on 21)

The binding member according to claim 21, comprising an LCDR3 having amino acid sequence SEQ ID NO: 120.

### Claim 23

An isolated binding member for human IL-6, comprising an HCDR3, an LCDR3 and/or a set of CDRs of any of antibodies 2, 3, 4, 5, 7, 8, 10, 14, 16, 17, 18, 19, 21, 22 or 23.

### Claim 24 (depends on 23)

The binding member according to claim 23, comprising a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein:
HCDR1 is SEQ ID NO: 3;
HCDR2 is SEQ ID NO: 4;
HCDR3 is SEQ ID NO: 115;
LCDR1 is SEQ ID NO: 8;
LCDR2 is SEQ ID NO: 9; and
LCDR3 is SEQ ID NO: 80.

### Claim 25 (depends on 23)

The binding member according to claim 23, wherein the binding member comprises an antibody molecule comprising an antibody VH domain and an antibody VL domain, wherein the antibody molecule comprises the set of CDRs, wherein the VH domain comprises HCDR1, HCDR2, HCDR3 and a framework and the VL domain comprises LCDR1, LCDR2, LCDR3 and a framework.

### Claim 26 (depends on 25)

The binding member according to claim 25, wherein the antibody molecule comprises an antibody constant region.

### Claim 27 (depends on 26)

The binding member according to claim 26, wherein the antibody molecule is an IgG1.

### Claim 28 (depends on 25)

A binding member according to claim 25, wherein the framework regions of the VH and/or VL domain are germlined to human germline gene segment sequences.

### Claim 29 (depends on 28)

The binding member according to claim 28, wherein the antibody VH domain comprises framework regions germlined to human germline framework Vh3_DP-86_(3-66).

### Claim 30 (depends on 29)

The binding member according to claim 29, wherein VH domain comprises framework regions FR1, FR2, FR3 and FR4 having amino acid sequences wherein FR1 is SEQ ID NO: 167, FR2 is SEQ ID NO: 168, FR3 is SEQ ID NO: 169 and FR4 is SEQ ID NO: 170.

### Claim 31 (depends on 30)

The binding member according to claim 30, wherein the antibody VH domain has the VH domain amino acid sequence shown in SEQ ID NO: 112.

### Claim 32 (depends on 28)

The binding member according to claim 28, wherein the antibody VL domain comprises framework regions germlined to human germline framework Vk1_L12.

### Claim 33 (depends on 32)

The binding member according to claim 32, wherein the VL domain comprises framework regions FR1, FR2, FR3 and FR4 having amino acid sequences wherein FR1 is SEQ ID NO: 171, FR2 is SEQ ID NO: 172, FR3 is SEQ ID NO: 173 and FR4 is SEQ ID NO: 174.

### Claim 34 (depends on 33)

The binding member according to claim 33, wherein the antibody VL domain has the VL domain amino acid sequence shown in SEQ ID NO: 117.

### Claim 35

An isolated antibody molecule comprising a heavy chain comprising amino acid sequence SEQ ID NO: 112 and a light chain comprising amino acid sequence SEQ ID NO: 117.

### Claim 36

An isolated antibody molecule that binds IL-6, wherein the antibody molecule comprises a VH domain amino acid sequence at least 90% identical to SEQ ID NO: 112 and a VL domain amino acid sequence at least 90% identical to SEQ ID NO: 117.

### Claim 37 (depends on 35, 36)

The antibody molecule according to claim 35 or claim 36, wherein the antibody molecule is an IgG.

### Claim 38 (depends on 37)

The antibody molecule according to claim 37, wherein the IgG is IgG1.

### Claim 39 (depends on 25)

The isolated VH domain of an antibody molecule according to claim 25.

### Claim 40 (depends on 25)

The isolated VL domain of an antibody molecule according to claim 25.

### Claim 41 (depends on 1, 36)

A composition comprising an isolated binding member according to claim 1, or an antibody molecule according to claim 36, and a pharmaceutically acceptable excipient.

### Claim at position 42

42-48. (canceled)

### Claim 49 (depends on 1, 36)

A method of treating a disorder associated with IL-6 in an individual, comprising administering a binding member according to claim 1, or an antibody molecule according to claim 36, to the individual.

### Claim 50 (depends on 49)

The method according to claim 49, wherein the disorder is an inflammatory and/or autoimmune disease.

### Claim 51 (depends on 50)

The method according to claim 50, wherein the disorder is rheumatoid arthritis, osteoarthritis, cachexia, chronic obstructive pulmonary disease, Juvenile idiopathic arthritis, asthma, systemic lupus erythematosus, inflammatory bowel disease, Crohn's disease or atherosclerosis.

### Claim 52 (depends on 51)

The method according to claim 51, wherein the disorder is rheumatoid arthritis.

### Claim 53 (depends on 49)

The method according to claim 49, wherein the disorder is a tumour and/or cancer.

### Claim 54 (depends on 1, 36, 39, 40)

An isolated nucleic acid molecule comprising a nucleotide sequence encoding a binding member according to claim 1, a VH domain according to claim 39, a VL domain according to claim 40, or an antibody molecule according to claim 36.

### Claim 55 (depends on 54)

A host cell in vitro transformed with nucleic acid according to claim 54.

### Claim 56 (depends on 55)

A method of producing a binding member, an antibody molecule or an antibody VH or VL domain, comprising culturing host cells according to claim 55 under conditions for production of the binding member, antibody molecule or antibody VH or VL domain.

### Claim 57 (depends on 56)

The method according to claim 56, further comprising isolating and/or purifying the binding member, antibody molecule, VH domain or VL domain.

### Claim 58 (depends on 56)

The method according to claim 56, further comprising formulating the binding member, antibody molecule, VH domain or VL domain into a composition comprising at least one additional component.

### Claim 59

A method for producing an antibody antigen-binding domain for IL-6, the method comprising
providing, by way of addition, deletion, substitution or insertion of one or more amino acids in the amino acid sequence of a parent VH domain comprising HCDR1, HCDR2 and HCDR3, wherein the parent VH domain HCDR1, HCDR2 and HCDR3 are a set of HCDRs as shown in Table 7, a VH domain which is an amino acid sequence variant of the parent VH domain, and optionally combining the VH domain thus provided with one or more VL domains to provide one or more VH/VL combinations; and
testing said VH domain which is an amino acid sequence variant of the parent VH domain or the VH/VL combination or combinations to identify an antibody antigen binding domain for IL-6.

### Claim 60 (depends on 59)

The method according to claim 59, wherein said one or more VL domains is provided by way of addition, deletion, substitution or insertion of one or more amino acids in the amino acid sequence of a parent VL domain comprising LCDR1, LCDR2 and LCDR3, wherein the parent VL domain LCDR1, LCDR2 and LCDR3 are a VL set of CDRs as shown in Table 7, producing one or more VL domains each of which is an amino acid sequence variant of the parent VL domain.

### Claim 61 (depends on 59)

The method according to claim 59, wherein said VH domain which is an amino acid sequence variant of the parent VH domain is provided by CDR mutagenesis.

### Claim 62 (depends on 59)

The method according to claim 59 further comprising producing the antibody antigen-binding domain as a component of an IgG, scFv or Fab antibody molecule.

### Claim 63

A method for producing a binding member that binds IL-6, which method comprises:
providing starting nucleic acid encoding a VH domain or a starting repertoire of nucleic acids each encoding a VH domain, wherein the VH domain or VH domains either comprise a HCDR1, HCDR2 and/or HCDR3 to be replaced or lack a HCDR1, HCDR2 and/or HCDR3 encoding region;
combining said starting nucleic acid or starting repertoire with donor nucleic acid or donor nucleic acids encoding or produced by mutation of the amino acid sequence of an HCDR1, HCDR2, and/or HCDR3 shown in Table 7, such that said donor nucleic acid is or donor nucleic acids are inserted into the CDR1, CDR2 and/or CDR3 region in the starting nucleic acid or starting repertoire, so as to provide a product repertoire of nucleic acids encoding VH domains;
expressing the nucleic acids of said product repertoire to produce product VH domains;
optionally combining said product VH domains with one or more VL domains;
selecting a binding member for IL-6, which binding member comprises a product VH domain and optionally a VL domain; and
recovering said binding member or nucleic acid encoding it.

### Claim 64 (depends on 63)

The method according to claim 63 wherein the donor nucleic acids are produced by mutation of said HCDR1 and/or HCDR2.

### Claim 65 (depends on 63)

The method according to claim 63 wherein the donor nucleic acid is produced by mutation of HCDR3.

### Claim 66 (depends on 63)

The method according to claim 63, comprising providing the donor nucleic acid by random mutation of nucleic acid.

### Claim 67 (depends on 59, 63)

The method according to claim 59 or 63, further comprising attaching a product VH domain that is comprised within the recovered binding member to an antibody constant region.

### Claim 68 (depends on 67)

The method according to claim 67 comprising providing an IgG, scFv or Fab antibody molecule comprising the product VH domain and a VL domain.

### Claim 69 (depends on 68)

The method according to claim 68, further comprising testing the antibody antigen-binding domain or binding member that binds IL-6 for ability to neutralize IL-6.

### Claim 70 (depends on 69)

The method according to claim 69, wherein a binding member that comprises an antibody molecule that binds and neutralizes IL-6 is obtained.

### Claim 71 (depends on 70)

The method according to claim 70, wherein the antibody molecule is an scFv.

### Claim 72 (depends on 70)

The method according to claim 70, wherein the antibody molecule is an IgG.

### Claim 73 (depends on 56)

A method for producing an antibody molecule composition, comprising obtaining an antibody molecule using a method according to claim 56, and formulating the antibody molecule into a composition comprising at least one additional component.