# US 11,078,290 B2 · Amino acid sequences directed against RANK-L and polypeptides comprising the same for the treatment of bone diseases and disorders

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

- Number: US 11,078,290 B2
- Document ID: US11078290B2
- Status: Granted
- Kind: B2
- Application number: 15/356,714
- Filed: 2016-11-21
- Published: 2017-06-15
- Granted: 2021-08-03
- Earliest claimed: 2007-05-24
- Assignees: Ablynx N.V.
- Applicants: Ablynx N.V.
- Inventors: Els Anna Alice Beirnaert; Sigrid Cornelis; Hendricus Renerus Jacobus Mattheus Hoogenboom
- Web page: https://patentgrep.com/patent/US11078290B2

## CPC

- A61P 19/00: Drugs for skeletal disorders
- C07K 16/2875: {against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 (against NGF C07K16/22, against TNF C07K16/241)}
- A61K 39/3955: {against proteinaceous materials, e.g. enzymes, hormones, lymphokines}
- A61K 47/60: the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
- A61K 47/6849: {the antibody targeting a receptor, a cell surface antigen or a cell surface determinant}
- A61P 19/08: for bone diseases, e.g. rachitism, Paget's disease
- A61P 19/10: for osteoporosis
- A61P 37/02: Immunomodulators
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 14/765: Serum albumin, e.g. HSA
- C07K 16/24: against cytokines, lymphokines or interferons
- A61K 2039/505: {comprising antibodies}
- C07K 2317/22: from camelids, e.g. camel, llama or dromedary
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C07K 2317/31: multispecific
- C07K 2317/35: Valency
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/567: Framework region [FR]
- C07K 2317/569: Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K 2317/94: Stability, e.g. half-life, pH, temperature or enzyme-resistance
- C07K 2319/31: fusions, other than Fc, for prolonged plasma life, e.g. albumin
- 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)
- 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

The present invention relates to amino acid sequences that are directed against RANK-L, as well as to compounds or constructs, and in particular proteins and polypeptides, that comprise or essentially consist of one or more such amino acid sequences. The invention also relates to nucleic acids encoding such amino acid sequences and polypeptides; to methods for preparing such amino acid sequences and polypeptides; to host cells expressing or capable of expressing such amino acid sequences or polypeptides; to compositions, and in particular to pharmaceutical compositions, that comprise such amino acid sequences, polypeptides, nucleic acids and/or host cells; and to uses of such amino acid sequences or polypeptides, nucleic acids, host cells and/or compositions, in particular for prophylactic, therapeutic or diagnostic purposes.

## Claims

### Claim 1

A method for inhibiting and/or preventing binding of RANK-L to RANK, wherein the method comprises at least the step of contacting RANK-L with at least one polypeptide that specifically binds to RANK-L, or with a composition comprising at least one polypeptide that specifically binds to RANK-L, wherein the at least one polypeptide comprises or essentially consists of 4 framework regions (FR1 to FR4 respectively) and 3 complementary determining regions (CDR1 to CDR3 respectively), in which:
CDR1 is SEQ ID NO: 190, CDR2 is SEQ ID NO: 314, and CDR3 is SEQ ID NO: 438;
CDR1 is SEQ ID NO: 193, CDR2 is SEQ ID NO: 317, and CDR3 is SEQ ID NO: 441;
CDR1 is SEQ ID NO: 196, CDR2 is SEQ ID NO: 320, and CDR3 is SEQ ID NO: 444;
or
CDR1 is SEQ ID NO: 205, CDR2 is SEQ ID NO: 329, and CDR3 is SEQ ID NO: 453.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the at least one polypeptide comprises or essentially consists of 4 framework regions (FR1 to FR4 respectively) and 3 complementary determining regions (CDR1 to CDR3 respectively), in which the CDR sequences have 100% amino acid identity with the CDR sequences of at least one of the amino acid sequences of SEQ ID NOs: 562, 565, 568, 577, 730-745, 757, and 765.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the at least one polypeptide essentially consists of a domain antibody, of a single domain antibody, of a “dAb”, a Nanobody, or a humanized Nanobody.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the at least one polypeptide is a VHH sequence, a partially humanized VHH sequence, a fully humanized VHH sequence, a camelized heavy chain variable domain or a Nanobody that has been obtained by affinity maturation.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the at least one polypeptide essentially consists of a Nanobody that i) has at least 80% amino acid identity with at least one of the amino acid sequences chosen from the group consisting of SEQ ID NOs: 1-22, 560-571, 573-621, 730-749, 757, and 765, in which for the purposes of determining the degree of amino acid identity, the amino acid residues that form the CDR sequences are disregarded;
and in which:
ii) one or more of the amino acid residues at positions 11, 37, 44, 45, 47, 83, 84, 103, 104 and 108 according to the Kabat numbering are
at position 11: L, M, S, V, or W;
at position 37: F, Y, H, I, L, or V;
at position 44: G, E, A, D, Q, R, S, or L;
at position 45: L, R, C, I, L, P, Q, or V;
at position 47: W, L, F, A, G, I, M, R, S, V, or Y;
at position 83: R, K, N, E, G, I, M, Q, or T;
at position 84: P, A, L, R, S, T, D, or V;
at position 103: W, P, R, or S;
at position 104: G or D; and
at position 108: Q, L, or R.

### Claim 6 (depends on 1)

The method according to claim 1, wherein the at least one polypeptide comprises or essentially consists of at least two Nanobodies that are chosen from the group consisting of SEQ ID NOs: 562, 565, 568, 577, 730-745, 757, and 765, or from the group consisting of amino acid sequences that have more than 80% sequence identity with at least one of the amino acid sequences of SEQ ID NOs: 562, 565, 568, 577, 730-745, 757, and 765.

### Claim 7 (depends on 1)

The method according to claim 1, wherein the at least one polypeptide is chosen from the group consisting of amino acid sequences that have more than 80% sequence identity with one or more of the amino acid sequences of SEQ ID NOs: 622-624, 626-630, 632-636, 638, 639, 646-648, 651-654, 656-660, 662-666, 668-672, 674, 675, 682-684, 687-690, 692-696, 698-702, 704-708, 710-711, 718-720, 723-726, 728, 729, 760, 762, 767-771, and 773.

### Claim 8 (depends on 1)

The method according to claim 1, wherein the at least one polypeptide
i) has at least 80% amino acid identity with at least one of the amino acid sequences of SEQ ID NOs: 622-624, 626-630, 632-636, 638, 639, 646-648, 651-654, 656-660, 662-666, 668-672, 674, 675, 682-684, 687-690, 692-696, 698-702, 704-708, 710-711, 718-720, 723-726, 729, 760, 762, 767-771, and 773, in which for the purposes of determining the degree of amino acid identity, the amino acid residues that form the CDR sequences are disregarded;
and in which:
in the Nanobodies that are encompassed in the polypeptide, one or more of the amino acid residues at positions 11, 37, 44, 45, 47, 83, 84, 103, 104 and 108 according to the Kabat numbering are
at position 11: L, M, S, V, or W;
at position 37: F, Y, H, I, L, or V;
at position 44: G, E, A, D, Q, R, S, or L;
at position 45: L, R, C, I, L, P, Q, or V;
at position 47: W, L, F, A, G, I, M, R, S, V, or Y;
at position 83: R, K, N, E, G, I, M, Q, or T;
at position 84: P, A, L, R, S, T, D, or V;
at position 103: W, P, R, or S;
at position 104: G or D; and
at position 108: Q, L, or R.

### Claim 9 (depends on 1)

The method according to claim 1, wherein the at least one polypeptide further comprises one or more other groups, residues, moieties or binding units.

### Claim 10 (depends on 9)

The method according to claim 9, in which said one or more other groups, residues, moieties or binding units provide the polypeptide with increased half-life.

### Claim 11 (depends on 10)

The method according to claim 10, in which said one or more other groups, residues, moieties or binding units that provide the polypeptide with increased half-life are chosen from the group consisting of serum proteins or fragments thereof, binding units that can bind to serum proteins, an Fc portion, and small proteins or peptides that can bind to serum proteins.

### Claim 12 (depends on 10)

The method according to claim 10, in which said one or more other binding units that provide the polypeptide with increased half-life are chosen from the group consisting of binding units that can bind to serum albumin and a serum immunoglobulin.

### Claim 13 (depends on 10)

The method according to claim 10, in which said one or more other binding units that provide the polypeptide with increased half-life are chosen from the group consisting of domain antibodies, single domain antibodies, “dAb”'s, and Nanobodies that can bind to serum albumin or a serum immunoglobulin.

### Claim 14 (depends on 1)

The method according to claim 1, wherein the at least one polypeptide modulates the differentiation and/or proliferation of osteoclasts.

### Claim 15

A polypeptide that specifically binds to RANK-L, wherein the polypeptide comprises or essentially consists of 4 framework regions (FR1 to FR4 respectively) and 3 complementary determining regions (CDR1 to CDR3 respectively), in which:
CDR1 is SEQ ID NO: 190, CDR2 is SEQ ID NO: 314, and CDR3 is SEQ ID NO: 438;
CDR1 is SEQ ID NO: 193, CDR2 is SEQ ID NO: 317, and CDR3 is SEQ ID NO: 441;
CDR1 is SEQ ID NO: 196, CDR2 is SEQ ID NO: 320, and CDR3 is SEQ ID NO: 444; or
CDR1 is SEQ ID NO: 205, CDR2 is SEQ ID NO: 329, and CDR3 is SEQ ID NO: 453.