Non-natural amatoxin-type antibody conjugate
Disclosed is a non-natural amatoxin-type antibody conjugate, said conjugate similar to the natural amatoxin is linked to a biopharmaceutically acceptable salt with a target biomolecule so as to obtain stability in blood plasma, and efficiently kill tumor cells in cells.
1. A non-natural amatoxin-antibody conjugate, comprising a toxin moiety having a structural formula (I) or a pharmaceutically acceptable salt thereof, and a biomacromolecule moiety A having a binding affinity to a target,
wherein:
R 1 is H, —OH or —O-L-A;
R 2 is H or —OH;
R 3 is OC 1-6 alkyl;
R 4 is H or -L-A-NH2 or —OH;
R 5 is —NH 2 , —OH, —NH-L-A or —O-L-A;
wherein O is an oxygen atom, N is a nitrogen atom and H is a hydrogen atom,
wherein A represents a biomacromolecule moiety, and
wherein L is a linker, comprising the following structure:
-L 1 -[L 2 ] m -[AA] n -L 3 -,
wherein: L 1 is a linker for connecting to A, and wherein L 1 is selected from:
wherein a wavy line connects to A;
L 2 is a spacer connecting L 1 to AA; m is an integer selected from 1-6; L 3 is a structural fragment connecting to the toxin moiety; AA is a structural fragment consisting of 1 to 6 amino acids, and n is 0 or 1.
2. The non-natural amatoxin-antibody conjugate claim 1 , wherein
R 1 is —OH;
R 2 is —OH;
R 3 is OCH 3 ;
R 4 is H;
R 5 is —NH-L-A, and
wherein L is
and
wherein the left-side wavy line connects to A, and the right-side wave line connects to NH in R 5 .
3. The non-natural amatoxin-antibody conjugate of claim 1 , wherein A comprises an antibody or its antigen-binding fragment thereof, or a nucleic acid aptamer.
4. The non-natural amatoxin-antibody conjugate of claim 3 , wherein the antibody or its antigen-binding fragment thereof-comprises a chimeric antibody, a deimmunized antibody, a humanized antibody, a human antibody, a bispecific antibody, a tetra-specific antibody, a nanobody or their antigen-binding fragments thereof.
5. The non-natural amatoxin-antibody conjugate of claim 3 , wherein the antigen-binding fragment is selected from the group consisting of Fab, F(ab′), Fd, Fv, a single-chain Fv or a disulfide-linked Fv(dsFv).
6. The non-natural amatoxin-antibody conjugate of claim 1 , wherein only one of R 1 , R 4 and R 5 comprises a -L-A structure.
7. The non-natural amatoxin-antibody conjugate of claim 1 , wherein
R 1 corresponds to —H or —OH;
R 2 corresponds to —H or —OH;
R 3 corresponds to —OC 1-6 alkyl;
R 4 corresponds to —NH 2 or —OH;
R 5 corresponds to —NH-L-A or —O-L-A.
8. The non-natural amatoxin-antibody conjugate of claim 1 , wherein L 2 is a substituted or unsubstituted C1-C6 alkyl, a substituted or unsubstituted C3-C20 cycloalkyl, a substituted or unsubstituted C3-C20 heterocycloalkyl, a substituted or unsubstituted C5-C20 aryl, a substituted or unsubstituted C5-C20 heteroaryl, or —(CH 2 CH 2 O)a-, wherein a is an integer selected from 1 to 20.
9. The non-natural amatoxin-antibody conjugate of claim 8 , wherein the L 2 substitution comprises a substitution with a hydroxyl group, a sulfydryl group, halogen, a carboxyl group, an amino group, a phosphate group, a nitro group, a cyano group, a sulfo group, or a substituted or unsubstituted C1-C6 alkyl.
10. The non-natural amatoxin-antibody of claim 9 , wherein the further substituted comprises a substitution with a hydroxyl group, a sulfydryl group, halogen, a carboxyl group, an amino group, a phosphate group, a nitro group, a cyano group, or a sulfo group.
11. The non-natural amatoxin-antibody conjugate of claim 1 , wherein the amino acids are L-amino acids selected from glycine, alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, serine, tyrosine, cysteine, methionine, asparagine, glutamine, threonine, aspartate, glutamate, lysine, arginine or histidine.
12. The non-natural amatoxin-antibody conjugate of claim 1 , wherein the amino acids are selected from phenylalanine, citrulline, valine, lysine, serine, glutamate, aspartate or glycine.
13. The non-natural amatoxin-antibody conjugate of claim 1 , wherein L 3 comprises
wherein a wavy line represents the connection to AA and an asterisk represents the connection to the toxin moiety,
wherein n is 1,
wherein L4 is a carbonyl group or a single bond,
wherein L5 is C2-C12 alkyl, and
wherein R6 and R7 are each independently selected from hydrogen or C1-C6 alkyl.
14. The non-natural amatoxin-antibody conjugate of claim 13 , wherein L4 is a carbonyl group.
15. A pharmaceutical composition comprising the non-natural amatoxin-type antibody conjugate of claim 1 and a pharmaceutically acceptable carrier.
16. A method of making the pharmaceutical composition of claim 15 , comprising combining the non-natural amatoxin-type antibody conjugate of claim 1 with the pharmaceutically acceptable carrier.
17. The pharmaceutical composition of claim 15 , wherein the pharmaceutical composition is an anti-lung cancer drug, an anti-renal cancer drug, an anti-urethral cancer drug, an anti-colorectal cancer drug, an anti-prostate cancer drug, an anti-glioblastoma drug, an anti-ovarian cancer drug, an anti-pancreatic cancer drug, an anti-breast cancer drug, an anti-melanoma drug, an anti-liver cancer drug, an anti-bladder cancer drug, an anti-malignant lymphoma drug, an anti-leukemia drug, an anti-gastric cancer drugs or an anti-esophageal cancer drug.