Granted Patent B2

US 12,318,454 · App. 18/315,869 · Granted Jun 3, 2025

Inventors: Daniel Paul Teufel (Cambridge, GB); Catherine Lucy Stace (Cambridge, GB); Silvia Pavan (Cambridge, GB); Edward Walker (Cambridge, GB); Leonardo Baldassarre (Cambridge, GB)

Assignee: BicycleRD Limited

A61K47/64A61K31/195A61K47/547A61K51/0482A61K51/08C07K7/02C07K7/08A61P35/00

Abstract

Claims (68)

1. A method of preparing a drug conjugate of formula (IV):

or a salt thereof, wherein:

bicycle represents a peptide ligand specific for MT1-MMP (membrane type 1 metalloprotease) comprising a polypeptide comprising at least three cysteine residues, separated by at least two loop sequences, and a molecular scaffold which forms covalent bonds with the cysteine residues of the polypeptide such that at least two polypeptide loops are formed on the molecular scaffold, wherein the peptide ligand comprises the amino acid sequence

-C i -X 1 -U/O 2 -X 3 -X 4 -G 5 -C ii -E 6 -D 7 -F 8 -Y 9 -X 10 -X 11 -C iii -(SEQ ID NO: 1), or

(β-Ala)-Sar10-AC(D-Ala)NE(1Nal)(D-Ala)CEDFYD(tBuGly)C(SEQ ID NO: 5);

wherein:

C i , C ii and C iii represent first, second and third cysteine residues, respectively;

each of X 1 , X 3 , X 4 , X 10 and X 11 represents any amino acid residue;

U/O 2 represents that position 2 of the loop sequence between C i and C ii is U or O;

U represents a polar, uncharged amino acid residue selected from the group consisting of N, C, Q, M, S and T; and

O represents a non-polar aliphatic amino acid residue selected from the group consisting of G, A, I, L, P and V;

toxin is the cytotoxic agent DM1;

R 3 represents methyl, and R 1 , R 2 and R 4 each represent hydrogen;

m represents 1; and

n represents 1,

the method comprising reacting a compound

or a salt thereof, with a compound

2. The method of claim 1 , wherein the compound

or a salt thereof, reacts with the compound

in a solution comprising about 50% volume dimethylacetamide, about 50% volume sodium acetate buffer at pH about 5.0, and EDTA.

3. The method of claim 1 , wherein X 1 is selected from the group consisting of Y, M, F and V.

4. The method of claim 1 , wherein U/O 2 represents N or G.

5. The method of claim 1 , wherein X 3 is U or Z, wherein (i) U is selected from the group consisting of N, C, Q, M, S and T, and Z is selected from the group consisting of D and E; or

(ii) U is Q and Z is E.

6. The method of claim 1 , wherein X 4 is selected from the group consisting of F, W and Y.

7. The method of claim 1 , wherein X 10 is selected from the group consisting of D and E; and X 11 is selected from the group consisting of G, A, I, L, P and V.

8. The method of claim 1 , wherein the peptide ligand comprises the amino acid sequence selected from the group consisting of:

-C i -Y-N-E-F-G-C ii -E-D-F-Y-D-I-C iii -(SEQ ID NO: 2),

βAla-Sar10-A-CYNEFGCEDFYDIC (SEQ ID NO: 3),

C(D-Ala)NEFGCEDFYDIC (SEQ ID NO: 4),

(β-Ala)-Sar10-AC(D-Ala)NE(1Nal)(D-Ala)CEDFYD(tBuGly)C SEQ ID NO: 5),

-C i -Y/M/F/V-U/O-U/Z-J-G-C ii -E-D-F-Y-Z-O-C iii -(SEQ ID NO: 6),

-C i -Y/M/F/V-N/G-E/Q-F-G-C ii -E-D-F-Y-D-I-C iii -(SEQ ID NO: 7),

-C i -Y/M/F-N/G-E/Q-F-G-C ii -E-D-F-Y-D-I-C iii -(SEQ ID NO: 8),

-C i -Y/M-N-E/Q-F-G-C ii -E-D-F-Y-D-I-C iii -(SEQ ID NO: 9),

-C i -M-N-Q-F-G-C ii -E-D-F-Y-D-I-C iii -(SEQ ID NO: 10);

-C i -F-G-E-F-G-C ii -E-D-F-Y-D-I-C iii -(SEQ ID NO: 11);

-C i -V-N-E-F-G-C ii -E-D-F-Y-D-I-C iii -(SEQ ID NO: 12);

-C i -F-N-E-F-G-C ii -E-D-F-Y-D-I-C iii -(SEQ ID NO: 13);

-C i -Y-N-E-Y-G-C ii -E-D-F-Y-D-I-C iii -(SEQ ID NO: 14); and

-C i -Y-N-E-W-G-C ii -E-D-F-Y-D-I-C iii -(SEQ ID NO: 15).

9. The method of claim 1 , wherein the molecular scaffold is TBMB (1,3,5-tris (bromomethyl) benzene), which forms covalent bonds with the cysteine residues of the polypeptide yielding a tri-substituted 1,3,5-trismethylbenzene structure.

10. A method of preparing a drug conjugate of formula (IV):

or a salt thereof, wherein:

bicycle represents a peptide ligand comprising:

a polypeptide of the amino acid sequence

(β-Ala)-Sar10-AC(D-Ala)NE(1Nal)(D-Ala)CEDFYD (tBuGly)C (SEQ ID NO: 5); and

a molecular scaffold which is TBMB (1,3,5-tris (bromomethyl) benzene), which forms covalent bonds with the cysteine residues of the polypeptide yielding a tri-substituted 1,3,5-trismethylbenzene structure;

toxin is the cytotoxic agent DM1;

R 3 represents methyl, and R 1 , R 2 and R 4 each represent hydrogen;

m represents 1; and

n represents 1,

the method comprising reacting a compound

or a salt thereof,

with a compound

in a solution comprising about 50% volume dimethylacetamide, about 50% volume sodium acetate buffer at pH about 5.0, and EDTA.

11. A compound

or a salt thereof, wherein bicycle represents a peptide ligand specific for MT1-MMP (membrane type 1 metalloprotease) comprising a polypeptide comprising at least three cysteine residues, separated by at least two loop sequences, and a molecular scaffold which forms covalent bonds with the cysteine residues of the polypeptide such that at least two polypeptide loops are formed on the molecular scaffold, wherein the peptide ligand comprises the amino acid sequence

-C i -X 1 -U/O 2 -X 3 -X 4 -G 5 -C ii -E 6 -D 7 -F 8 -Y 9 -X 10 -X 11 -C iii -(SEQ ID NO: 1), or

(β-Ala)-Sar10-AC(D-Ala)NE(1Nal)(D-Ala)CEDFYD(tBuGly)C SEQ ID NO: 5);

wherein:

C i , C ii and C iii represent first, second and third cysteine residues, respectively;

each of X 1 , X 3 , X 4 , X 10 and X 11 represents any amino acid residue;

U/O 2 represents that position 2 of the loop sequence between C i and C ii is U or O;

U represents a polar, uncharged amino acid residue selected from the group consisting of N, C, Q, M, S and T; and

O represents a non-polar aliphatic amino acid residue selected from the group consisting of G, A, I, L, P and V.

12. The compound of claim 11 , or a salt thereof, wherein the peptide ligand comprises the amino acid sequence (β-Ala)-Sar10-AC (D-Ala) NE(1Nal) (D-Ala)CEDFYD(tBuGly)C(SEQ ID NO: 5).

13. The compound of claim 11 , or a salt thereof, wherein the molecular scaffold is TBMB (1,3,5-tris (bromomethyl) benzene), which forms covalent bonds with the cysteine residues of the polypeptide yielding a tri-substituted 1,3,5-trismethylbenzene structure.

Assignments (2)

Priority Claims (2)

Continuity (4)

References Cited (400)