Tubulysins and protein-tubulysin conjugates

A compound having the following formula

• or a pharmaceutically acceptable salt thereof, wherein
• BA is a binding agent;
• L is a linker covalently bound to BA and to T;
• T is
• wherein
• X is —O— or —NR5;
• R5 is hydrogen, —CH3—, —(CH2)2—OH, —(CH2)2—NH2, —CH2—C(O)—OH, —(CH2)2—O—(CH2)2—NH2, —(CH2CH2—O)2—(CH2)2—NH2, or —C(O)—CH2—NH2;
• R2 is —O—C(O)—NH—CH2—CH(OH)—CH2OH, —NH—C(O)CH3, —O—CH2CH3, —O—(CH2)3—OH, —O—C(O)CH3, —O—C(O)—NH—(CH2)2—OH, or —O—C(O)—NH—(CH2CH2O)2—(CH2)2OH;
• R3 is —OH, —NH2, —NH—(CH2)2OH, —NH—CH2—C(O)—NH2, —NH—CH2—C(O)—OH, —NH—C(O)—CH2NH2, —NH—[(CH2)2OH]—C(O)—NH2, —NH—CH2—(CH2O)2—(CH2)2—NH2, or —NH—CH2—C(O)—NH—(CH2CH2O)2—(CH2)2NH;
• R4 is hydrogen or —F;
• R7 when present is —CH3;
• R6 is —OH or —NH—C(O)OH;
• Q is —CH2— or —O—;
• R1 is —C1-C8 alkyl, —C1-C8 alkenyl, or —C1-C8 alkynyl;
• r is three or four;
• and
• k is an integer from one to thirty;
• wherein T is not a compound selected from the Table below
• ID.
• Structure
• P1
• P2
• P3
• P4
• P5
• P6
• P7
• P8
• P9
• P10
• P12
• P13
• P14
• P15
• P16
• P17
• P18
• P19
• P20
• P21
• P22
• P25
• P26
• P27
• P28
• P31
• P32
• P34
• P35
• P36
• P51
• IVq
• IVu
• IVvA
• IVvB
• Vb
• Ve
• IX
• X
• D-5a
• or a pharmaceutically acceptable salt thereof, covalently bound to L.

A compound having the structure of Formula (I)

• or a pharmaceutically acceptable salt thereof, wherein
• X is —O— or —NR5,
• R5 is a hydrogen, —CH3, —(CH2)2—OH, —(CH2)2—NH2, —CH2—C(O)—OH, —(CH2)2—O—(CH2)2—NH2, —(CH2CH2—O)2—(CH2)2—NH2, or —C(O)—CH2—NH2;
• R2 is —O—C(O)—NH—CH2—CH(OH)—CH2OH, —NH—C(O)CH3, —O—CH2CH3, —O—(CH2)3—OH, —O—C(O)CH3, —O—C(O)—NH—(CH2)2—OH, —O—C(O)—NH—(CH2CH2O)3—CH2NH—C(O)CH2NH2, or —O—C(O)—NH—(CH2CH2O)2—(CH2)2OH;
• R3 is —OH, —NH2, —NH—(CH2)2OH, —NH—CH2—C(O)—NH2, —NH—CH2—C(O)—OH, —NH—C(O)—CH2NH2, —NH—[(CH2)2OH]—C(O)—NH2, —NH—CH2—(CH2O)2—(CH2)2—NH2, —N(CH2CH2OH)(C(O)CH2NH2), or —NH—CH2—C(O)—NH—(CH2CH2O)2—(CH2)2NH2;
• R4 is hydrogen or —F
• R7 when present is —CH3;
• R6 is —OH, —NHCH2C(O)OH, or —NH—C(O)OH;
• Q is —CH2— or —O—;
• R1 is —C5 alkyl or —C5 alkynyl;
• r is three or four;
• and
• wherein T is not a compound selected from the table below
• ID.
• Structure
• P2
• P5
• P6
• P7
• P10
• P12
• P13
• P14
• P15
• P16
• P17
• P18
• P19
• P20
• P21
• P22
• P25
• P26
• P27
• P28
• P31
• P32
• P34
• P35
• P36
• P51
• IVq
• IVu
• IVvA
• IVvB
• Vb
• Ve
• IX
• X
• and
• D-5a

A linker-payload having the formula

• L-T
• or a pharmaceutically acceptable salt thereof, wherein
• L is a linker covalently bound to T;
• T is
• covalently bound to L, wherein
• X is —O— or —NR5;
• R5 is hydrogen, —CH3, —(CH2)2—OH, —(CH2)2—NH2, —CH2—C(O)—OH, —(CH2)2—O—(CH2)2—NH2, —(CH2CH2—O)2—(CH2)2—NH2, —C(O)—CH2—NH2, a first N-terminal amino acid residue, a first amino acid residue;
• or
• R5 is a covalent bond to L;
• or
• R5 is —(CH2)2—OH or —CH2—C(O)—OH with a covalent bond to L from the terminal oxygen in —(CH2)2—OH or —CH2—C(O)—OH;
• or
• R5 is —(CH2)2—NH2, —(CH2)2—O—(CH2)2—NH2, —(CH2CH2—O)2—(CH2)2—NH2, —C(O)—CH2—NH2, a first N-terminal amino acid residue, or a first amino acid residue with a covalent bond to L from the nitrogen in any one of —(CH2)2—NH2,
• —(CH2)2—O—(CH2)2—NH2, —(CH2CH2—O)2—(CH2)2—NH2, or —C(O)—CH2—NH2, the first N-terminal amino acid residue, or the first amino acid residue;
• R2 is —O—C(O)—NH—CH2—CH(OH)—CH2OH, —NH—C(O)CH3, —O—CH2CH3, —O—(CH2)3—OH, —O—C(O)CH3, —O—C(O)—NH—(CH2)2—OH, —O—C(O)—NH—(CH2CH2O)3—CH2NH—C(O)CH2NH2, or —O—C(O)—NH—(CH2CH2O)2—(CH2)2OH;
• or
• R2 is —O—C(O)—NH—CH2—CH(OH)—CH2OH, —O—(CH2)3—OH, —O—C(O)—NH—(CH2)2—OH, or —O—C(O)—NH—(CH2CH2O)2—(CH2)2OH with a covalent bond to L from a terminal oxygen in any one of —O—C(O)—NH—CH2—CH(OH)—CH2OH, —O—(CH2)3—OH, —O—C(O)—NH—(CH2)2—OH, or —O—C(O)—NH—(CH2CH2O)2—(CH2)2OH;
• or
• R2 is —O—C(O)—NH—(CH2CH2O)3—CH2NH—C(O)CH2NH2 with a covalent bond to L from the terminal nitrogen in —O—C(O)—NH—(CH2CH2O)3—CH2NH—C(O)CH2NH2;
• R3 is —OH, —NH2, —NH—(CH2)2OH, —NH—CH2—C(O)—NH2, —NH—CH2—C(O)—OH, —NH—C(O)—CH2NH2, —NH—[(CH2)2OH]—C(O)—NH2, —NH—CH2—(CH2O)2—(CH2)2—NH2, —N(CH2CH2OH)(C(O)CH2NH2), or —NH—CH2—C(O)—NH—(CH2CH2O)2—(CH2)2NH2;
• or
• R3 is —OH, —NH—(CH2)2OH, —NH—CH2—C(O)—OH, or —N(CH2CH2OH)(C(O)CH2NH2) with a covalent bond to L from a terminal oxygen in any one of —OH, —NH—(CH2)2OH, —NH—CH2—C(O)—OH, or —N(CH2CH2OH)(C(O)CH2NH2);
• or
• R3 is —NH2, —NH—CH2—C(O)—NH2, —NH—C(O)—CH2NH2, —NH—[(CH2)2OH]—C(O)—NH2, —NH—CH2—(CH2O)2—(CH2)2—NH2, —N(CH2CH2OH)(C(O)CH2NH2), or —NH—CH2—C(O)—NH—(CH2CH2O)2—(CH2)2NH2 with a covalent bond to L from a terminal nitrogen in any one of —NH2, —NH—CH2—C(O)—NH2, —NH—C(O)—CH2NH2, —NH—[(CH2)2OH]—C(O)—NH2, —NH—CH2—(CH2O)2—(CH2)2—NH2, —N(CH2CH2OH)(C(O)CH2NH2), or —NH—CH2—C(O)—NH—(CH2CH2O)2—(CH2)2NH2;
• R4 is hydrogen or —F;
• R7 when present is —CH3;
• R6 is —OH, —NHCH2C(O)OH, or —NH—C(O)OH;
• or
• R6 is —OH, —NHCH2C(O)OH, or —NH—C(O)OH with a covalent bond to L from the terminal oxygen in any one of —OH, —NHCH2C(O)OH, or —NH—C(O)OH;
• Q is —CH2— or —O—;
• R1 is —C5 alkyl, —C6 alkyl or —C5 alkynyl;
• r is three or four;
• and
• k is an integer from one to thirty;
• wherein T is not a compound selected from the table below
• ID.
• Structure
• P2
• P5
• P6
• P7
• P10
• P12
• P13
• P14
• P15
• P16
• P17
• P18
• P19
• P20
• P21
• P22
• P25
• P26
• P27
• P28
• P31
• P32
• P34
• P35
• P36
• P51
• IVq
• IVu
• IVvA
• IVvB
• Vb
• Ve
• IX
• X
• and
• D-5a