# US 2023/0272084 A1 · B7-h3 directed antibody drug conjugates

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- Number: US 2023/0272084 A1
- Document ID: US20230272084A1
- Status: Published application
- Kind: A1
- Application number: 18/151,146
- Filed: 2023-01-06
- Published: 2023-08-31
- Earliest claimed: 2016-04-15
- Assignees: MacroGenics, Inc.
- Applicants: MacroGenics, Inc.
- Inventors: Deryk T. LOO; Ling HUANG; Leslie S. JOHNSON; Thomas SON; Juniper A. SCRIBNER; Ezio BONVINI
- Web page: https://patentgrep.com/patent/US20230272084A1

## CPC

- C07K 16/2827: {against B7 molecules, e.g. CD80, CD86}
- A61K 39/395: Antibodies (agglutinins A61K38/36 {; as drug carriers A61K47/50}); Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K 47/6803: {Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates}
- A61K 47/68031: {the drug being an auristatin}
- A61K 47/6811: {the drug being a protein or peptide, e.g. transferrin or bleomycin}
- A61K 47/6851: {the antibody targeting a determinant of a tumour cell}
- A61K 47/6879: {the immunoglobulin having two or more different antigen-binding sites, e.g. bispecific or multispecific immunoglobulin}
- A61P 35/00: Antineoplastic agents
- A61K 2039/505: {comprising antibodies}
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C07K 2317/55: Fab or Fab'
- C07K 2317/77: Internalization into the cell
- 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

The present invention is directed to novel B7-H3-binding molecules capable of binding to human and non-human B7-H3, and in particular to such molecules that are cross-reactive with B7-H3 of a non-human primate (e.g., a cynomolgus monkey). The invention additionally pertains to B7-H3-binding molecules that comprise Variable Light Chain and/or Variable Heavy Chain (VH) Domains that have been humanized and/or deimmunized so as to exhibit a reduced immunogenicity upon administration to recipient subjects. The invention particularly pertains to bispecific, trispecific or multispecific B7-H3-binding molecules, including bispecific diabodies, BiTEs, bispecific antibodies, trivalent binding molecules, etc. that comprise: (i) such B7-H3-binding Variable Domains and (ii) a domain capable of binding to an epitope of a molecule present on the surface of an effector cell. The invention is also directed to pharmaceutical compositions that contain any of such B7-H3-binding molecules, and to methods involving the use of any of such B7-H3-binding molecules in the treatment of cancer and other diseases and conditions. The invention also particularly pertains to a molecule that comprises the human B7-H3 binding domain of a humanized anti-human B7-H3 antibody conjugated to at least one drug moiety (a “B7-H3-ADC”). The invention is also directed to pharmaceutical compositions that contain such B7-H3-ADCs, and to methods involving the use of any of such B7-H3-ADCs in the treatment of cancer and other diseases and conditions.

## Claims

### Claim at position 1

1-32. (canceled)

### Claim 33

An anti B7-H3 antibody drug conjugate (B7-H3-ADC) that comprises the formula:
Ab-(LM)m-(D)n,
wherein:
Ab is a humanized B7-H3 antibody or B7-H3 binding fragment thereof that binds to B7-H3 and comprises:
(i) the CDRL1 sequence RASESIYSYLA, the CDRL2 sequence NTKTLPE and the CDRL3 sequence QHHYGTPPWT in its Variable Light Chain (VL) domain; and
(ii) the CDRH1 sequence SYGMS, the CDRH2 sequence TINSGGSNTYYPDSLKG and the CDRH3 sequence HDGGAMDY in its Variable Heavy Chain (VH) domain;
D is a cytotoxic drug moiety that comprises a duocarmycin cytotoxin;
LM comprises at least one bond or a Linker Molecule that covalently links Ab and D;
m is an integer between 0 and n and denotes the number of Linker Molecules of the B7-H3-ADC;
and
n is an integer between 1 and 10 and denotes the number of cytotoxic drug moieties covalently linked to the B7-H3-ADC molecule.

### Claim 34 (depends on 33)

The B7-H3-ADC of claim 33, wherein said Ab comprises:
(i) a humanized Variable Light Chain (VL) domain comprising the amino acid sequence of SEQ ID NO: 20; and
(ii) a humanized Variable Heavy Chain (VH) domain comprising the amino acid sequence of SEQ ID NO: 21;

### Claim 35 (depends on 34)

The B7-H3-ADC of claim 34, wherein said Ab further comprises an Fe domain of a human IgG.

### Claim 36 (depends on 35)

The B7-H3-ADC of claim 35, wherein said human IgG is a human IgG1, IgG2, IgG3, or IgG4.

### Claim 37 (depends on 35)

The B7-H3-ADC of claim 35, wherein said human IgG is a human IgG1.

### Claim 38 (depends on 35)

The B7-H3-ADC of claim 35, wherein said Fc Domain is a variant Fc Domain that comprises:
(a) one or more amino acid modifications that reduces the affinity of the variant Fc Domain for an FcγR; and/or
(b) one or more amino acid modifications that enhances the serum half-life of the variant Fc Domain.

### Claim 39 (depends on 38)

The B7-H3-ADC of claim 38, wherein said modifications that reduce the affinity of the variant Fc Domain for an FcγR comprise the substitution of L234A; L235A; or L234A and L235A, wherein the numbering is that of the EU index as in Kabat.

### Claim 40 (depends on 38)

The B7-H3-ADC of claim 38, wherein said modifications that enhance the serum half-life of the variant Fc Domain comprise the substitution of M252Y; M252Y and S254T; M252Y and T256E; M252Y, S254T and T256E; or K288D and H435K, wherein the numbering is that of the EU index as in Kabat.

### Claim 41 (depends on 33)

The B7-H3-ADC of claim 33, wherein at least one of said LM moieties is a Linker Molecule.

### Claim 42 (depends on 41)

The B7-H3-ADC of claim 41, wherein said LM Linker Molecule comprises a cleavable linker.

### Claim 43 (depends on 42)

The B7-H3-ADC of claim 42, wherein said cleavable linker is a peptidic linker.

### Claim 44 (depends on 43)

The B7-H3-ADC of claim 43, wherein said peptidic linker is a valine-citrulline dipeptide linker.

### Claim 45 (depends on 42)

The B7-H3-ADC of claim 42, wherein said LM Linker Molecule further comprises a self-eliminating spacer between the cleavable linker and D.

### Claim 46 (depends on 45)

The B7-H3-ADC of claim 45, wherein said self-eliminating spacer comprises a para-aminobenzyloxycarbonyl moiety.

### Claim 47 (depends on 42)

The B7-H3-ADC of claim 42, wherein said Linker Molecule further comprises a maleimide linker moiety between the cleavable linker and Ab.

### Claim 48 (depends on 40)

The B7-H3-ADC of claim 40, wherein LM comprises the formula:
[V-(W)k-(X)1-A]
and said conjugate comprises the formula:
Ab-[V-(W)k-(X)1-A]-D
wherein:
V is a cleavable linker,
(W)k-(X)1-A is an elongated, self-eliminating spacer system, that self-eliminates via a 1, (4+2n)-elimination,
W and X are each a 1, (4+2n) electronic cascade spacer, being the same or different,
A is either a spacer group of formula (Y)m, wherein Y is a 1, (4+2n) electronic cascade spacer, or a group of formula U, being a cyclisation elimination spacer,
k, l and m are independently an integer of 0 (included) to 5 (included),
n is an integer of 0 (included) to 10 (included),
with the provisos that:
when A is (Y)m: then k+l+m≥1, and
if k+l+m=1, then n>l;
when A is U: then k+1≥1,
W, X, and Y are independently selected from compounds having the formula:
or the formula:
wherein: Q is —R5C═CR6—, S, O, NR5, —R5C═N—, or —N═CR5—
P is NR7, O or S
a, b, and c are independently an integer of 0 (included) to 5 (included);
I, F and G are independently selected from compounds having the formula:
wherein R1, R2, R3, R4, R5, R6, R7, R8, and R9 independently represent H, C1-6 alkyl, C3-20 heterocyclyl, C5-20 aryl, C1-6 alkoxy, hydroxy (OH), amino (NH2), mono-substituted amino (NRxH), di-substituted amino (NRx1Rx2), nitro (NO2), halogen, CF3, CN, CONH2, SO2Me, CONHMe, cyclic C1-5 alkylamino, imidazolyl, C1-6 alkylpiperazinyl, morpholino, thiol (SH), thioether (SRx), tetrazole, carboxy (COOH), carboxylate (COORx), sulphoxy (S(═O)2OH), sulphonate (S(═O)2ORx), sulphonyl (S(═O)2Rx), sulphixy (S(═O)OH), sulphinate (S(═O)ORx), sulphinyl (S(═O)Rx), phosphonooxy (OP(═O)(OH)2), and phosphate (OP(═O)(ORx)2), where Rx, Rx1 and Rx2 are independently selected from a C1-6 alkyl group, a C3-20 heterocyclyl group or a C5-20 aryl group, two or more of the substituents R1, R2, R3, R4, R5, R6, R7, R8, or R9 optionally being connected to one another to form one or more aliphatic or aromatic cyclic structures;
U is selected from compounds having the formula:
wherein:
a, b and c are independently selected to be an integer of 0 or 1;
provided that a +b+c=2 or 3;
R1 and/or R2 independently represent H, C1-6 alkyl, the alkyl being optionally substituted with one or more of the following groups: hydroxy (OH), ether (ORx), amino (NH2), mono-substituted amino (NRxH), disubstituted amino (NRx1Rx2), nitro (NO2), halogen, CF3, CN, CONH2, SO2Me, CONHMe, cyclic C1-5 alkylamino, imidazolyl, C1-6 alkylpiperazinyl, morpholino, thiol (SH), thioether (SRx), tetrazole, carboxy (COOH), carboxylate (COORx), sulphoxy (S(═O)2OH), sulphonate (S(═O)2ORx), sulphonyl (S(═O)2Rx), sulphixy (S(═O)OH), sulphinate (S(═O)ORx), sulphinyl (S(═O)Rx), phosphonooxy (OP(═O)(OH)2), and phosphate (OP(═O)(ORx)2), where Rx, Rx1 and Rx2 are selected from a C1-6 alkyl group, a C3-20 heterocyclyl group or a C5-20 aryl group; and
R3, R4, R5, R6, R7 and R8 independently represent H, C1-6 alkyl, C3-20 heterocyclyl, C5-20 aryl, C1-6 alkoxy, hydroxy (OH), amino (NH2), mono-substituted amino (NRxH), disubstituted amino (NRx1Rx2), nitro (NO2), halogen, CF3, CN, CONH2, SO2Me, CONHMe, cyclic C1-5 alkylamino, imidazolyl, C1-6 alkylpiperazinyl, morpholino, thiol (SH), thioether (SRx), tetrazole, carboxy (COOH), carboxylate (COORx), sulphoxy (S(═O)2OH), sulphonate (S(═O)2ORx), sulphonyl (S(═O)2Rx), sulphixy (S(═O)OH), sulphinate (S(═O)ORx), sulphinyl (S(═O)Rx), phosphonooxy (OP(═O)(OH)2), and phosphate (OP(═O)(ORx)2), where Rx, Rx1 and Rx2 are selected from a C1-6 alkyl group, a C3-20 heterocyclyl group or a C5-20 aryl group, and two or more of the substituents R1, R2, R3, R4, R5, R6, R7, or R8 are optionally connected to one another to form one or more aliphatic or aromatic cyclic structures.

### Claim 49 (depends on 48)

The B7-H3-ADC of claim 48, wherein said LM linker molecule comprises:
(1) p-aminobenzyloxycarbonyl-p-aminobenzyloxycarbonyl;
(2) p-aminobenzyloxycarbonyl-p-aminobenzyloxycarbonyl-p-aminobenzyloxycarbonyl;
(3) p-ammocinnamyloxycarbonyl;
(4) p-aminocinnamyloxycarbonyl-p-aminobenzyloxycarbonyl;
(5) p-amino-benzyloxycarbonyl-p-aminocinnamyloxycarbonyl;
(6) p-aminocinnamyloxycarbonyl-p-aminocinnamyloxycarbonyl;
(7) p-aminophenylpentadienyloxycarbonyl;
(8) p-aminophenylpentadienyloxycarbonyl-p-arninocinnamyloxycarbonyl;
(9) p-aminophenylpentadienyloxycarbonyl-paminobenzyloxycarbonyl;
(10) p-aminophenylpentadienyloxycarbonyl-p-aminophenylpentadienyloxycarbonyl;
(11) p-aminobenzyloxycarbonyl(methylamino)ethyl(methylamino) carbonyl;
(12) p-aminocinnamyloxycarbonyl(methylamino)ethyl(methylamino) carbonyl;
(13) p-aminobenzyloxycarbonyl-p-aminobenzyloxycarbonyl(methylamino) ethyl(methylamino)carbonyl;
(14) p-aminocinnamyloxycarbonyl-p-aminobenzyloxycarbonyl (methylamino)ethyl(methylamino)carbonyl;
(15) p-aminobenzyloxycarbonyl-p-arninocinnamyloxycarbonyl (methylamino)ethyl(methylamino)-carbonyl;
(16) p-aminocinnamyloxycarbonyl-p-aminocinnamyloxycarbonyl (methylamino)ethyl(methylamino)carbonyl;
(17) p-aminobenzyloxycarbonyl-p-aminobenzyl;
(18) p-aminobenzyloxycarbonyl-p-aminobenzyloxycarbonyl-p-aminobenzyl;
(19) p-aminocinnamyl;
(20) p-aminocinnamyloxycarbonyl-p-aminobenzyl;
(21) p-aminobenzyloxycarbonyl-p-aminocinnamyl;
(22) p-amino-cinnamyloxycarbonyl-p-aminocinnamyl;
(23) p-aminophenylpentadienyl;
(24) p-aminophenylpentadienyloxycarbonyl-p-aminocinnamyl;
(25) p-aminophenylpentadienyloxycarbonyl-p-aminobenzyl; or
(26) p-aminophenylpentadienyloxycarbonyl-p-aminophenylpentadienyl.

### Claim 50 (depends on 41)

The B7-H3-ADC of claim 41, wherein said LM Linker Molecule is conjugated to the side chain of an amino acid of a polypeptide chain of Ab and binds the Ab to a molecule of the cytotoxic drug moiety D.

### Claim 51 (depends on 33)

The B7-H3-ADC of claim 33, wherein said duocarmycin cytotoxin is selected from the group consisting of: duocarmycin A, duocarmycin B1, duocarmycin B2, duocarmycin C1, duocarmycin C2, duocarmycin D, duocarmycin SA, CC-1065, adozelesin, bizelesin, carzelesin (U-80244), seco-duocarmycin and spiro-duocarmycin (DUBA).

### Claim 52 (depends on 51)

The B7-H3-ADC of claim 51, wherein said duocarmycin cytotoxin is seco-duocarmycin.

### Claim 53 (depends on 41)

The B7-H3-ADC of claim 41, wherein said LM Linker Molecule is covalently linked to the Ab via reduced inter-chain disulfides.

### Claim 54 (depends on 33)

The B7-H3-ADC of claim 33, wherein said antibody or said B7-H3 binding fragment thereof comprises a human IgG Constant Domain Kappa Domain.

### Claim 55 (depends on 54)

The B7-H3-ADC of claim 54, wherein said human IgG Constant Domain Kappa Domain comprises the amino acid sequence of SEQ ID NO:1.

### Claim 56 (depends on 55)

The B7-H3-ADC of claim 55, wherein:
Ab is said antibody and said antibody comprises:
(i) a light chain comprising the Variable Light Chain (VL) domain comprising the amino acid sequence of SEQ ID NO:20 and a CL Kappa Domain of SEQ ID NO:1; and
(ii) a heavy chain comprising the Variable Heavy Chain (VH) domain comprising the amino acid sequence of SEQ ID NO:21, a CH1 Domain of SEQ ID NO:3, a Hinge Domain of SEQ ID NO:7 and a Fc Domain comprising a CH2-CH3 Domain of SEQ ID NO:12; and
said LM comprises a Linker Molecule comprising a maleimide linker moiety, a valine-citrulline dipeptide linker, and a para-aminobenzyloxycarbonyl moiety.

### Claim 57 (depends on 33)

A pharmaceutical composition comprising an effective amount of the B7-H3-ADC of claim 33 and a pharmaceutically acceptable carrier, excipient or diluent.

### Claim 58 (depends on 57)

A pharmaceutical composition comprising an effective amount of the B7-H3-ADC of claim 57 and a pharmaceutically acceptable carrier, excipient or diluent.