# US 9,751,946 B2 · Anti-CLL-1 antibodies and immunoconjugates

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

- Number: US 9,751,946 B2
- Document ID: US9751946B2
- Status: Granted
- Kind: B2
- Application number: 14/852,369
- Filed: 2015-09-11
- Published: 2016-03-17
- Granted: 2017-09-05
- Earliest claimed: 2014-09-12
- Assignees: Genentech, Inc.
- Applicants: Genentech, Inc.
- Inventors: Bing Zheng; Andrew Polson; Cecilia Chiu; Wei-Ching Liang; Yan Wu
- Web page: https://patentgrep.com/patent/US9751946B2

## CPC

- C07K 16/2851: {against the lectin superfamily, e.g. CD23, CD72}
- A61K 31/5517: condensed with five-membered rings having nitrogen as a ring hetero atom, e.g. imidazobenzodiazepines, triazolam
- A61K 31/706: containing six-membered rings with nitrogen as a ring hetero atom
- A61K 47/68: the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K 47/6817: {Toxins}
- A61K 47/6867: {the tumour determinant being from a cell of a blood cancer}
- A61K 47/6889: {Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment}
- A61K 51/1069: {the tumor cell being from blood cells, e.g. the cancer being a myeloma}
- A61K 51/1093: {conjugates with carriers being antibodies}
- A61P 35/00: Antineoplastic agents
- A61P 35/02: specific for leukemia
- C07K 16/3061: {Blood cells}
- G01N 33/57505: of the blood, e.g. leukaemia
- A61K 2039/505: {comprising antibodies}
- C07K 2317/14: Specific host cells or culture conditions, e.g. components, pH or temperature
- 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 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
- C07K 2317/52: Constant or Fc region; Isotype
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K 2317/77: Internalization into the cell
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- C07K 2317/94: Stability, e.g. half-life, pH, temperature or enzyme-resistance
- G01N 2333/7056: Selectin superfamily, e.g. LAM-1, GlyCAM, ELAM-1, PADGEM
- 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
- 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 invention provides anti-CLL-1 antibodies and immunoconjugates and methods of using the same.

## Claims

### Claim 1

An isolated monoclonal anti-CLL-1 antibody, wherein the antibody comprises
(a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:8;
(b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:11;
(c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:10;
(d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:5;
(e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:6; and
(f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:7.

### Claim 2 (depends on 1)

The antibody of claim 1, wherein the antibody comprises one or more engineered free cysteine amino acid residues.

### Claim 3 (depends on 2)

The antibody of claim 2, wherein the one or more engineered free cysteine amino acid residues is located in the light chain.

### Claim 4 (depends on 3)

The antibody of claim 3, wherein the one or more engineered free cysteine amino acid residues in the light chain comprises K149C according to Kabat numbering.

### Claim 5 (depends on 1)

The antibody of claim 1, wherein the antibody is a humanized antibody.

### Claim 6 (depends on 1)

The antibody of claim 1, wherein the antibody is an antibody fragment that binds CLL-1.

### Claim 7 (depends on 1)

The antibody of claim 1, wherein the antibody is an IgG1, IgG2a, or IgG2b antibody.

### Claim 8 (depends on 1)

The antibody of claim 1, wherein the antibody is an IgG1 antibody.

### Claim 9 (depends on 4)

The antibody of claim 4, wherein the antibody is an IgG1 antibody.

### Claim 10 (depends on 1)

The antibody of claim 1, wherein the antibody is conjugated to a label.

### Claim 11 (depends on 10)

The antibody of claim 10, wherein the label is a positron emitter.

### Claim 12 (depends on 11)

The antibody of claim 11, wherein the positron emitter is 89Zr.

### Claim 13

An isolated monoclonal anti-CLL-1 antibody, wherein the antibody comprises (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34 and (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO:32.

### Claim 14 (depends on 13)

The antibody of claim 13, wherein the antibody comprises one or more engineered free cysteine amino acid residues.

### Claim 15 (depends on 14)

The antibody of claim 14, wherein the one or more engineered free cysteine amino acid residues is located in the light chain.

### Claim 16 (depends on 15)

The antibody of claim 15, wherein the one or more engineered free cysteine amino acid residues in the light chain comprises K149C according to Kabat numbering.

### Claim 17 (depends on 13)

The antibody of claim 13, wherein the antibody is a humanized antibody.

### Claim 18 (depends on 13)

The antibody of claim 13, wherein the antibody is an antibody fragment that binds CLL-1.

### Claim 19 (depends on 13)

The antibody of claim 13, wherein the antibody is an IgG1, IgG2a, or IgG2b antibody.

### Claim 20 (depends on 13)

The antibody of claim 13, wherein the antibody is an IgG1 antibody.

### Claim 21 (depends on 16)

The antibody of claim 16, wherein the antibody is an IgG1 antibody.

### Claim 22 (depends on 13)

The antibody of claim 13, wherein the antibody is conjugated to a label.

### Claim 23 (depends on 22)

The antibody of claim 22, wherein the label is a positron emitter.

### Claim 24 (depends on 23)

The antibody of claim 23, wherein the positron emitter is 89Zr.

### Claim 25 (depends on 1)

An immunoconjugate comprising:
the antibody of claim 1 and
a cytotoxic agent.

### Claim 26 (depends on 1)

An immunoconjugate having the formula Ab-(L-D)p, wherein:
(a) Ab is the antibody of claim 1;
(b) L is a linker that links the Ab to D;
(c) D is a cytotoxic drug; and
(d) p ranges from 1-8.

### Claim 27 (depends on 26)

The immunoconjugate of claim 26, wherein the cytotoxic drug is selected from a maytansinoid, a calicheamicin, a pyrrolobenzodiazepine, and a nemorubicin derivative.

### Claim 28 (depends on 26)

The immunoconjugate of claim 26, wherein D is a pyrrolobenzodiazepine of Formula A:
wherein the dotted lines indicate the optional presence of a double bond between C1 and C2 or C2 and C3;
R2 is selected from H, OH, ═O, ═CH2, CN, R, OR, ═CH—RD, ═C(RD)2, O—SO2—R, CO2R and COR, and optionally further selected from halo or dihalo, wherein RD is independently selected from R, CO2R, COR, CHO, CO2H, and halo;
R6 and R9 are independently selected from H, R, OH, OR, SH, SR, NH2, NHR, NRR′, NO2, Me3Sn and halo;
R7 is selected from H, R, OH, OR, SH, SR, NH2, NHR, NRR′, NO2, Me3Sn and halo;
Q is selected from O, S and NH, and RH is either H or R; or Q is O and RH is SO3M, wherein M is a metal cation;
R and R′ are each independently selected from optionally substituted C1-8 alkyl, C3-8 heterocyclyl and C5-20 aryl groups, and optionally in relation to the group NRR′, R and R′ together with the nitrogen atom to which they are attached form an optionally substituted 4-, 5-, 6- or 7-membered heterocyclic ring;
R12, R16, R19 and R17 are as defined for R2, R6, R9 and R7 respectively;
R″ is a C3-12 alkylene group, which chain may be interrupted by one or more heteroatoms and/or aromatic rings that are optionally substituted; and
X and X′ are independently selected from O, S and NH.

### Claim 29 (depends on 26)

The immunoconjugate of claim 26, wherein D has the structure:
wherein n is 0 or 1.

### Claim 30 (depends on 26)

The immunoconjugate of claim 26, wherein D has the structure:

### Claim 31 (depends on 26)

The immunoconjugate of claim 26, wherein D is a nemorubicin derivative.

### Claim 32 (depends on 31)

The immunoconjugate of claim 31, wherein D has the structure:

### Claim 33 (depends on 30)

The immunoconjugate of claim 30, wherein the immunoconjugate has the structure:

### Claim 34 (depends on 33)

The immunoconjugate of claim 33, wherein p ranges from 2-5.

### Claim 35 (depends on 33)

The immunoconjugate of claim 33, wherein p is 1.

### Claim 36 (depends on 33)

The immunoconjugate of claim 33, wherein p is 2.

### Claim 37 (depends on 33)

The immunoconjugate of claim 33, wherein Ab comprises an engineered free cysteine amino acid residue located in the light chain at K149C according to Kabat numbering and wherein Ab is an IgG1 antibody.

### Claim 38 (depends on 26)

A pharmaceutical formulation comprising the immunoconjugate of claim 26 and a pharmaceutically acceptable carrier.

### Claim 39 (depends on 38)

The pharmaceutical formulation of claim 38, further comprising an additional therapeutic agent.

### Claim 40 (depends on 39)

The pharmaceutical formulation of claim 39, wherein the additional therapeutic agent is selected from 5-azacytidine and decitabine.

### Claim 41 (depends on 40)

The pharmaceutical formulation of claim 40, wherein the additional therapeutic agent is 5-azacytidine.

### Claim 42 (depends on 40)

The pharmaceutical formulation of claim 40, wherein the additional therapeutic agent is decitabine.

### Claim 43 (depends on 13)

An immunoconjugate comprising:
the antibody of claim 13 and
a cytotoxic agent.

### Claim 44 (depends on 13)

An immunoconjugate having the formula Ab-(L-D)p, wherein:
(a) Ab is the antibody of claim 13;
(b) L is a linker that links the Ab to D;
(c) D is a cytotoxic drug; and
(d) p ranges from 1-8.

### Claim 45 (depends on 44)

The immunoconjugate of claim 44, wherein the cytotoxic drug is selected from a maytansinoid, a calicheamicin, a pyrrolobenzodiazepine, and a nemorubicin derivative.

### Claim 46 (depends on 44)

The immunoconjugate of claim 44, wherein D is a pyrrolobenzodiazepine of Formula A:
wherein the dotted lines indicate the optional presence of a double bond between C1 and C2 or C2 and C3;
R2 is selected from H, OH, ═O, ═CH2, CN, R, OR, ═CH—RD, ═C(RD)2, O—SO2—R, CO2R and COR, and optionally further selected from halo or dihalo, wherein RD is independently selected from R, CO2R, COR, CHO, CO2H, and halo;
R6 and R9 are independently selected from H, R, OH, OR, SH, SR, NH2, NHR, NRR′, NO2, Me3Sn and halo;
R7 is selected from H, R, OH, OR, SH, SR, NH2, NHR, NRR′, NO2, Me3Sn and halo;
Q is selected from O, S and NH, and R″ is either H or R; or Q is 0 and R″ is SO3M, wherein M is a metal cation;
R and R′ are each independently selected from optionally substituted C1-8 alkyl, C3-8 heterocyclyl and C5-20 aryl groups, and optionally in relation to the group NRR′, R and R′ together with the nitrogen atom to which they are attached form an optionally substituted 4-, 5-, 6- or 7-membered heterocyclic ring;
R12, R16, R19 and R17 are as defined for R2, R6, R9 and R7 respectively;
R″ is a C3-12 alkylene group, which chain may be interrupted by one or more heteroatoms and/or aromatic rings that are optionally substituted; and
X and X′ are independently selected from O, S and NH.

### Claim 47 (depends on 44)

The immunoconjugate of claim 44, wherein D has the structure:
wherein n is 0 or 1.

### Claim 48 (depends on 44)

The immunoconjugate of claim 44, wherein D has the structure:

### Claim 49 (depends on 44)

The immunoconjugate of claim 44, wherein D is a nemorubicin derivative.

### Claim 50 (depends on 49)

The immunoconjugate of claim 49, wherein D has the structure:

### Claim 51 (depends on 48)

The immunoconjugate of claim 48, wherein the immunoconjugate has the structure:

### Claim 52 (depends on 51)

The immunoconjugate of claim 51, wherein p ranges from 2-5.

### Claim 53 (depends on 51)

The immunoconjugate of claim 51, wherein p is 1.

### Claim 54 (depends on 51)

The immunoconjugate of claim 51, wherein p is 2.

### Claim 55 (depends on 51)

The immunoconjugate of claim 51, wherein Ab comprises an engineered free cysteine amino acid residue located in the light chain at K149C according to Kabat numbering and wherein Ab is an IgG1 antibody.

### Claim 56 (depends on 44)

A pharmaceutical formulation comprising the immunoconjugate of claim 44 and a pharmaceutically acceptable carrier.

### Claim 57 (depends on 56)

The pharmaceutical formulation of claim 56, further comprising an additional therapeutic agent.

### Claim 58 (depends on 57)

The pharmaceutical formulation of claim 57, wherein the additional therapeutic agent is selected from 5-azacytidine and decitabine.

### Claim 59 (depends on 58)

The pharmaceutical formulation of claim 58, wherein the additional therapeutic agent is 5-azacytidine.

### Claim 60 (depends on 58)

The pharmaceutical formulation of claim 58, wherein the additional therapeutic agent is decitabine.

### Claim 61

An immunoconjugate comprising a structure:
wherein Ab is an isolated monoclonal anti-CLL-1 antibody, wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:8; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:11; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:10; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:5; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:6; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:7;
wherein the antibody comprises an engineered free cysteine residue located in the light chain at K149C according to Kabat numbering; and
wherein p is 2.

### Claim 62 (depends on 61)

The immunoconjugate of claim 61, wherein the antibody is a humanized antibody.

### Claim 63 (depends on 62)

The immunoconjugate of claim 62, wherein the antibody is an IgG1, IgG2a, or IgG2b antibody.

### Claim 64 (depends on 63)

The immunoconjugate of claim 63, wherein the antibody is an IgG1 antibody.

### Claim 65 (depends on 61)

A pharmaceutical formulation comprising the immunoconjugate of claim 61 and a pharmaceutically acceptable carrier.

### Claim 66 (depends on 65)

The pharmaceutical formulation of claim 65, further comprising an additional therapeutic agent.

### Claim 67 (depends on 66)

The pharmaceutical formulation of claim 66, wherein the additional therapeutic agent is selected from 5-azacytidine and decitabine.

### Claim 68 (depends on 67)

The pharmaceutical formulation of claim 67, wherein the additional therapeutic agent is 5-azacytidine.

### Claim 69 (depends on 67)

The pharmaceutical formulation of claim 67, wherein the additional therapeutic agent is decitabine.

### Claim 70

An immunoconjugate comprising a structure:
wherein Ab is an isolated monoclonal anti-CLL-1 antibody, wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:8; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:11; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:10; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:5; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:6; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:7;
wherein the antibody comprises an engineered free cysteine residue located in the light chain at K149C according to Kabat numbering; and
wherein p is 1.

### Claim 71 (depends on 70)

The immunoconjugate of claim 70, wherein the antibody is a humanized antibody.

### Claim 72 (depends on 71)

The immunoconjugate of claim 71, wherein the antibody is an IgG1, IgG2a, or IgG2b antibody.

### Claim 73 (depends on 72)

The immunoconjugate of claim 72, wherein the antibody is an IgG1 antibody.

### Claim 74 (depends on 70)

A pharmaceutical formulation comprising the immunoconjugate of claim 70 and a pharmaceutically acceptable carrier.

### Claim 75 (depends on 74)

The pharmaceutical formulation of claim 74, further comprising an additional therapeutic agent.

### Claim 76 (depends on 75)

The pharmaceutical formulation of claim 75, wherein the additional therapeutic agent is selected from 5-azacytidine and decitabine.

### Claim 77 (depends on 76)

The pharmaceutical formulation of claim 76, wherein the additional therapeutic agent is 5-azacytidine.

### Claim 78 (depends on 76)

The pharmaceutical formulation of claim 76, wherein the additional therapeutic agent is decitabine.

### Claim 79

An immunoconjugate comprising a structure:
wherein Ab is an isolated monoclonal anti-CLL-1 antibody, wherein the antibody comprises (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34 and (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO:32;
wherein the antibody comprises an engineered free cysteine residue located in the light chain at K149C according to Kabat numbering; and
wherein p is 2.

### Claim 80 (depends on 79)

The immunoconjugate of claim 79, wherein the antibody is a humanized antibody.

### Claim 81 (depends on 80)

The immunoconjugate of claim 80, wherein the antibody is an IgG1, IgG2a, or IgG2b antibody.

### Claim 82 (depends on 81)

The immunoconjugate of claim 81, wherein the antibody is an IgG1 antibody.

### Claim 83 (depends on 79)

A pharmaceutical formulation comprising the immunoconjugate of claim 79 and a pharmaceutically acceptable carrier.

### Claim 84 (depends on 83)

The pharmaceutical formulation of claim 83, further comprising an additional therapeutic agent.

### Claim 85 (depends on 84)

The pharmaceutical formulation of claim 84, wherein the additional therapeutic agent is selected from 5-azacytidine and decitabine.

### Claim 86 (depends on 85)

The pharmaceutical formulation of claim 85, wherein the additional therapeutic agent is 5-azacytidine.

### Claim 87 (depends on 85)

The pharmaceutical formulation of claim 85, wherein the additional therapeutic agent is decitabine.

### Claim 88

An immunoconjugate comprising a structure:
wherein Ab is an isolated monoclonal anti-CLL-1 antibody, wherein the antibody comprises (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34 and (b) a light chain variable region comprising the amino acid sequence of SEQ ID NO:32;
wherein the antibody comprises an engineered free cysteine residue located in the light chain at K149C according to Kabat numbering; and
wherein p is 1.

### Claim 89 (depends on 88)

The immunoconjugate of claim 88, wherein the antibody is a humanized antibody.

### Claim 90 (depends on 89)

The immunoconjugate of claim 89, wherein the antibody is an IgG1, IgG2a, or IgG2b antibody.

### Claim 91 (depends on 90)

The immunoconjugate of claim 90, wherein the antibody is an IgG1 antibody.

### Claim 92 (depends on 88)

A pharmaceutical formulation comprising the immunoconjugate of claim 88 and a pharmaceutically acceptable carrier.

### Claim 93 (depends on 92)

The pharmaceutical formulation of claim 92, further comprising an additional therapeutic agent.

### Claim 94 (depends on 93)

The pharmaceutical formulation of claim 93, wherein the additional therapeutic agent is selected from 5-azacytidine and decitabine.

### Claim 95 (depends on 94)

The pharmaceutical formulation of claim 94, wherein the additional therapeutic agent is 5-azacytidine.

### Claim 96 (depends on 94)

The pharmaceutical formulation of claim 94, wherein the additional therapeutic agent is decitabine.