The ADC of claim 13, wherein the unnatural amino acid is 3-formyltyrosine and the hydroxylamine group of the linker forms an oxime with the 3-formyl group of the unnatural amino acid.ġ5. The ADC of claim 1, wherein the linker has a structure as depicted in structure 4 or 5 before being coupled to the unnatural amino acid, wherein D is a drug moiety:ġ4. The ADC of claim 11, wherein the hydroxylamine of structure 2 or 3 is conjugated to the unnatural amino acid.ġ3. The ADC of claim 1, wherein the linker has a structure as depicted in structure 2 or 3 before being coupled to the unnatural amino acid: wherein Z is is selected from the group consisting of substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heteroalkenyl and substituted or unsubstituted heteroalkynyl wherein D is one or more drug moieties and wherein Y is a cleavage site such as a cleavage site for a cathepsin such as cathepsin B.ġ2. The ADC of claim 9, wherein the hydroxylamine of structure 1 is conjugated to the unnatural amino acid.ġ1.
The ADC of claim 1, wherein the linker has a structure as depicted in structure 1 before being coupled to the unnatural amino acid: wherein R is one or more drug moieties, which are optionally coupled to the hydroxylamine of structure 1 by one or more cleavage sites.ġ0. The ADC of claim 1, wherein Brentuximab is conjugated to two, four, six, or eight drug moieties.ĩ. The ADC of claim 1, wherein the linker comprises a hydroxylamine group and the unnatural amino acid comprises a formyl group ortho of a hydroxyl group in an aromatic ring, and wherein the hydroxylamine group of the linker forms an oxime with the formyl group of the unnatural amino acid after conjugation.Ĩ. The ADC of claim 1, wherein the linker is cleavable, such as by a protease like cathepsin B or wherein the linker comprises a valine-citrulline moiety.ħ. The ADC of claim 1, wherein the unnatural amino acid is a 2-substituted, 3-substituted or 4-substituted tyrosine or a tyrosine derivative substituted at the benzylic position or wherein the unnatural amino acid is 3-nitrotyrosine, 3-aminotyrosine, 3-azidotyrosine, 3-formyltyrosine, 3-acetyltyrosine, or 4-aminophenylalanine.Ħ. 4) or wherein the recognition sequence is VDSVEGEGEEEGEE (SEQ ID No. The ADC of claim 1, wherein the recognition sequence for tubulin tyrosine ligase has at least the amino acid sequence X 1X 2X 3X 4 (SEQ ID NO: 3), wherein X 1 and X 2 is any amino acid, X 3 is E, D or C and X 4 is E or wherein X 2 is G, S, A, V, or F and/or wherein X 1 is E, D, A, K, or P or wherein the recognition sequence is EGEE (SEQ ID No. The ADC of claim 1, wherein the drug moiety is selected from the group consisting of camptothecins, maytansinoids, calicheamycins, duocarmycins, tubulysins, amatoxins, dolastatins and auristatins such as monomethyl auristatin E (MMAE), pyrrolobenzodiazepine dimers, indolino-benzodiazepine dimers, radioisotopes, therapeutic proteins and peptides (or fragments thereof), nucleic acids, PROTACs, kinase inhibitors, MEK inhibitors, KSP inhibitors, and analogues or prodrugs thereof.Ĥ. The ADC of claim 1, wherein the heavy chains of Brentuximab have an amino acid sequence that comprises or consists of SEQ ID NO: 1 or have a sequence identity of at least 95% to SEQ ID NO: 1 and/or wherein the light chains of Brentuximab have an amino acid sequence that comprises or consists of SEQ ID NO: 2 or have a sequence identity of at least 95% to SEQ ID NO: 2 or wherein Brentuximab consists of heavy chains consisting of the amino acid sequence of SEQ ID NO: 1 and light chains consisting of the amino acid sequence of SEQ ID NO: 2.ģ. An antibody-drug conjugate (ADC) comprising: (a) Brentuximab, wherein Brentuximab comprises at the C-terminus of the light chains, the heavy chains or all of the heavy and light chains of the Brentuximab a recognition sequence for tubulin tyrosine ligase and a non-natural amino acid and (b) at least one drug moiety wherein a drug moiety is coupled to each of the non-natural amino acids via a linker.Ģ.