Drug-conjugates with a targeting molecule and two different drugs
There is disclosed an improved ADC (antibody drug conjugate) type composition having at least two different drug payloads conjugated to a single targeting protein. More specifically, the present disclosure attaches a first drug conjugate to a dual Cysteine residue on a targeting protein and a second drug conjugate with a different drug to a Lys residue on the targeting protein.
1. A method of preparing a dual-drug conjugate, the method comprising conjugating a first moiety comprising D 1 to a lysine side chain of A and conjugating a second moiety comprising D 2 to cysteine side chains of A, thereby producing a dual-drug conjugate which is a compound of Formula I
or a pharmaceutically acceptable salt thereof,
wherein:
A is an antibody or antigen-binding antibody fragment;
D 1 is an active agent;
each L 1 is independently a linker comprising at least one N (nitrogen) atom;
the nitrogen between the antibody or antigen-binding antibody fragment and the carbonyl of C(O)-L 1 -D 1 is the nitrogen of a Lys side chain of the antibody or antigen-binding antibody fragment;
D 2 an active agent, wherein D 1 and D 2 are different active agents;
each L 2 is independently a linker;
the E-component is selected from the group consisting of
L 3 is —(CH 2 )—;
L 4 is —(CH 2 )—;
m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and
n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
2. The method of claim 1 , wherein the first moiety is conjugated before conjugation of the second moiety.
3. The method of claim 1 , wherein the first moiety is conjugated after conjugation of the second moiety.
4. The method of claim 1 , wherein conjugating the first moiety to the lysine side chain of A comprises reacting a compound of formula I-A with a compound of formula I-B to provide a compound of formula I-C:
wherein G is —F, —Cl, —Br, —I, —N 3 , —OR, SR, —ONRR, RC(═O)O—, or RSO 2 —O—; and R is optionally substituted alkyl, or optionally substituted aryl.
5. The method of claim 1 , wherein conjugating the second moiety to the cysteine side chains of A comprises reacting a compound of formula I-D with a compound of formula I-E to provide a compound of formula I-F:
wherein each X is —Cl, —Br, —I, or RSO 2 —O—; and R is optionally substituted alkyl, or optionally substituted aryl.
6. The method of claim 5 , wherein the cysteine side chains were reduced with a reducing agent to prepare compound I-D.
7. The method of claim 1 , wherein conjugating the second moiety to the cysteine side chains of A comprises reacting a compound of formula II-A with a compound of formula I-E to provide a compound of formula II-B:
wherein each X is —Cl, —Br, —I, or RSO 2 —O—; and R is optionally substituted alkyl, or optionally substituted aryl.
8. The method of claim 1 , wherein conjugating the first moiety to the lysine side chain of A comprises reacting a compound of formula II-A with a compound of formula I-B to provide a compound of formula I-F:
wherein G is —F, —Cl, —Br, —I, —N 3 , —OR, SR, —ONRR, RC(═O)O—, or RSO 2 —O—; and R is optionally substituted alkyl, or optionally substituted aryl.
9. The method of claim 1 ,
wherein:
is:
10. The method of claim 1 , wherein L 1 comprises
where X 1 is N (nitrogen) or CH; Y 1 is N (nitrogen), or CH; and p is 0, 1, or 2.
11. The method of claim 1 , wherein L 1 comprises
12. The method of claim 1 , wherein D 1 is a tubulin binder, DNA alkylator, DNA intercalator, enzyme inhibitor, immune modulator, peptide, or nucleotide.
13. The method of claim 1 , wherein D 2 is a tubulin binder, DNA alkylator, DNA intercalator, enzyme inhibitor, immune modulator, peptide, or nucleotide.
14. The method of claim 1 , wherein L 1 or L 2 comprises —(CH 2 ) n — where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
15. The method of claim 1 , wherein L 1 or L 2 includes —(CH 2 CH 2 O) n — where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
16. The method of claim 1 , wherein the
S-linked portion of
comprises a modified L-Alanine residue.
17. The method of claim 1 , wherein each
is:
wherein G is —NH—.
18. The method of claim 1 , wherein each
is:
19. The method of claim 1 , wherein the dual-drug conjugate is:
or a pharmaceutically acceptable salt thereof, wherein A is an antibody or antigen-binding antibody fragment.
20. The method of claim 1 , wherein A is an antibody.
21. The method of claim 1 , wherein A is an antigen-binding antibody fragment.