Isoquinolidinobenzodiazepines

1. A compound having a structure of Formula (I) or (II):

wherein:

the dotted bond shown between —C(R a )— and —N(R b )— or —C(R a′ )— and —N(R b′ )— is independently a single bond or a double bond;

when a double bond is present between —C(R a )— and —N(R b )—, the —C(R a )— is olefinic and has a substituent R a and R b of the —N(R b )— is not present;

when a single bond is present between —C(R a )— and —N(R b )—, the —C(R a )— is saturated and has a hydrogen substituent in addition to the R a substituent and R b of the —N(R b )— is present;

when a double bond is present between —C(R a′ )— and —N(R b′ )—, the —C(R a′ )— is olefinic and has a substituent R a′ and R b′ of the —N(R b′ )— is not present;

when a single bond is present between —C(R a′ )— and —N(R b′ )—, the —C(R a′ )— is saturated and has a hydrogen substituent in addition to the R a′ substituent and R b′ of the —N(R b′ )— is present;

each of R a and R a′ is independently H or OH;

if present, each of R b and R b′ is independently H, L-R x or L-S c ;

R 2 , R 2′ , R 3 , R 3′ , R 4 , R 4′ , R 6′ and R 6 are each independently selected from H, OH, C 1 -C 10 alkyl, C 2 -C 10 alkenyl or C 2 -C 10 alkynyl;

each of R 5 or R 5′ is independently NH 2 , CO 2 H, H, OH, C 1 -C 10 alkyl, C 2 -C 10 alkenyl, C 2 -C 10 alkynyl, -L-R x or -L-S c ;

each of R 7 and R 7′ is H;

R 8 is:

H, NH 2 , CO 2 H, -L-R x , or -L-S c , wherein the carbon to which R 8 is attached also has a hydrogen substituent; or

an exo olefin having the structure

wherein the carbon to which R 8 is attached has no other substituent;

X is methylene;

each of Y and Y′ is independently O, S, or NH;

each of Z and Z′ is independently H, R, OH, OR, SH, SR, NH 2 , or NHR, where each R is independently unsubstituted C 1 -C 12 alkyl

L-R x is a linker L attached to a reactive moiety R x ;

-L-S c is a linker L attached to a substance S c ;

wherein:

L is a linker comprising the structure —HN-PEG-C(O)—X AA —, wherein PEG has 1-50 ethylene glycol units, and X AA is an amino acid sequence;

R x is a reactive moiety selected from the group consisting of a iodoacetamide, selenocysteine, succinimidyl, maleimidyl, cyclooctynyl, aminooxy, bisulfonyl, sulfonyl, and isothiocyanate moiety;

S c is a target binding agent selected from a protein, a portion of a protein, a peptide or a nucleic acid; and

when -L-R x or -L-S c is present in the compound of formula I or II, only one of R b , R b′ , R 5 , R 5′ , R 8 is L-R x or -L-S c ; and

wherein said compound is enantiomerically pure.

2. An antibody-drug conjugate having a structure of Formula III:

wherein:

is an antibody or antibody fragment that binds to a cancer marker on an AML cell;

W—R M is a linking moiety formed by W and R x , wherein W is a moiety attached to a natural or unnatural amino acid residue of the antibody/antibody fragment and R x is a reactive moiety linking L-IQB to the antibody, wherein R x is a succinimidyl, maleimidyl, cylooctynyl, aminooxy, bisulfonyl, sulfonyl, or isothiocyanate moiety, such that W—R M is a disulfide, a thiolated succinimidyl, an amino substituted succinimidyl, a (cyclooctyl)-1, 4 triazolyl, oxime substituted N-glycan, oxime, a substituted bis-sulfopropyl, a sulfonamidyl, an amide, or a thiocarbamate moiety;

L is a linker comprising the structure —HN-PEG-C(O)—X AA —, wherein PEG has 1-50 ethylene glycol units, and X AA is an amino acid sequence;

j is a number of 1 to 10; and,

IQB is a compound having a structure of Formula (I) or (II):

wherein:

the dotted bond shown between —C(R a )— and —N(R b )— or —C(R a′ )— and —N(R b′ )— is independently a single bond or a double bond;

when a double bond is present between —C(R a )— and —N(R b )—, the —C(R a )— is olefinic and has a substituent R a and R b of the —N(R b )— is not present;

when a single bond is present between —C(R a )— and —N(R b )—, the —C(R a )— is saturated and has a hydrogen substituent in addition to the R a substituent and R b of the —N(R b )— is present;

when a double bond is present between —C(R a′ )— and —N(R b′ )—, the —C(R a′ )— is olefinic and has a substituent R a′ and R b′ of the —N(R b′ )— is not present;

when a single bond is present between —C(R a′ )— and —N(R b′ )—, the —C(R a′ )— is saturated and has a hydrogen substituent in addition to the R a′ substituent and R b′ of the —N(R b′ )— is present;

each of R a and R a′ is independently H, OH, or —O—P, where P is a protecting group;

if present, each of R b and R b′ is independently H, or -L;

R 2 , R 2′ , R 3 , R 3′ , R 4 , R 4′ , R 6′ and R 6 are each independently selected from H, OH, C 1 -C 10 alkyl, C 2 -C 10 alkenyl or C 2 -C 10 alkynyl;

each of R 5 or R 5′ is independently NH 2 , CO 2 H, H, OH C 1 -C 10 alkyl, C 2 -C 10 alkenyl, C 2 -C 10 alkynyl, or -L;

each of R′ and R 7′ is H;

R 8 is:

H, NH 2 , CO 2 H, or -L, wherein the carbon to which R 8 is attached also has a hydrogen substituent; or

an exo olefin having the structure

wherein the carbon to which R 8 is attached has no other substituent;

X is:

C 1-12 alkylene, optionally wherein the alkylene chain is interrupted by one or more hetero atoms selected from the group consisting of O, S, and NH; or

—(CH 2 ) m -Q-(CH 2 ) p —, wherein m and p are each independently 0, 1 or 2;

Q has a structure of formula:

wherein each of R 9 , R 10 and R 11 is H, NH 2 , CO 2 H, -L; and

J is CH or N;

each of Y and Y′ is independently O, S, or NH;

each of Z and Z′ is independently H, R, OH, OR, SH, SR, NH 2 , or NHR, where each R is independently unsubstituted C 1 -C 12 alkyl; and

wherein only one of R b , R b′ , R 5 , R 5′ , R 8 , R 9 , R 10 , and R 11 is -L; and

wherein said IQB compound is enantiomerically pure.

3. The antibody-drug conjugate of claim 2 , having a structure of Formula III:

wherein:

is an antibody or antibody fragment that binds to a cancer marker on a AML cell;

W—R M is a linking moiety formed by W and R x , wherein W is a moiety attached a natural or unnatural amino acid residue of the antibody/antibody fragment and R x is a reactive moiety linking L-IQB to the antibody, wherein R x is a succinimidyl, maleimidyl, cylooctynyl, aminooxy, bisulfonyl, sulfonyl, or isothiocyanate moiety, such that W—R M is a disulfide, a thiolated succinimidyl, an amino substituted succinimidyl, a (cyclooctyl)-1, 4 triazolyl, oxime substituted N-glycan, oxime, a substituted bis-sulfopropyl, a sulfonamidyl, an amide, or a thiocarbamate moiety;

L is a linker comprising the structure —NH-PEG 8 -C(O)-VAL-ALA;

j is a number of 1 to 10; and,

IQB is a compound having a structure of Formula (I) or (II):

wherein:

the dotted bond shown between —C(R a )— and —N(R b )— or —C(R a′ )— and —N(R b′ )— is independently a single bond or a double bond;

each of R a and R a′ is independently H or OH;

each of R b and R b′ is not present, or is independently H, or a bond linked to linker L;

R 2 , R 2′ , R 3 , R 3′ , R 4 , R 4′ , R 6′ and R 6 are each independently selected from H, OH, C 1 -C 10 alkyl, C 2 -C 10 alkenyl or C 2 -C 10 alkynyl;

each of R 5 or R 5′ is independently NH 2 , CO 2 H, H, OH C 1 -C 10 alkyl, C 2 -C 10 alkenyl, C 2 -C 10 alkynyl, or a bond linked to linker L;

each of R 7 and R 7′ is H;

R 8 is:

H, NH 2 , CO 2 H, or a bond linked to linker L, wherein the carbon to which R 8 is attached also has a hydrogen substituent; or

an exo olefin having the structure

wherein the carbon to which R 8 is attached has no other substituent;

X is methylene;

each of Y and Y′ is independently O, S, or NH;

each of Z and Z′ is independently H, R, OH, OR, SH, SR, NH 2 , or NHR, where each R is independently unsubstituted C 1 -C 12 alkyl and

wherein only one of R b , R b′ , R 5 , R 5′ , R 8 is a bond linked to linker L; and

wherein said IQB compound is enantiomerically pure.

4. The antibody-drug conjugate of claim 2 , wherein W is attached directly or indirectly to the amino acid residue of the antibody/antibody fragment.

5. The antibody-drug conjugate of claim 2 , wherein R x is a succinimidyl, maleimidyl, cylooctynyl, aminooxy, bisulfonyl, sulfonyl, or isothiocyanate moiety.

6. The antibody-drug conjugate of claim 2 , wherein W—R M is a disulfide, a thiolated succinimidyl, an amino substituted succinimidyl, a (cyclooctyl)-1, 4 triazolyl, oxime substituted N-glycan, oxime, a substituted bis-sulfopropyl, a sulfonamidyl, an amide, or a thiocarbamate moiety.

7. The antibody-drug conjugate of claim 2 , wherein IQB is a moiety having the structure:

8. The antibody-drug conjugate of claim 2 , wherein —W-RM-L-IQB is a moiety having a structure of Formula IV:

9. The antibody-drug conjugate of claim 2 , wherein the antibody/antibody fragment is humanized.

10. The antibody-drug conjugate of claim 2 , wherein the antibody/antibody fragment is modified to introduce a non-natural cysteine residue.

11. The antibody-drug conjugate of claim 2 , wherein the antibody or antibody fragment is anti-CLL1.

12. The antibody-drug conjugate of claim 2 , wherein the antibody/antibody fragment is anti-CLL1 and IQB is a moiety having the structure:

13. The antibody-drug conjugate of claim 2 , wherein

the antibody or antibody fragment is anti-CLL1;

L is a linker comprising the structure:

W—R M is a thiolated succinimidyl; and

IQB is a moiety having the structure:

14. The antibody-drug conjugate of claim 2 , wherein the drug is conjugated to the non-natural cysteine residue.

15. The antibody-drug conjugate of claim 2 , wherein j is 1 to 3.

16. A pharmaceutical composition comprising a conjugate of claim 2 and a pharmaceutically acceptable carrier.

17. A method of treating acute myeloid leukemia (AML) comprising administering to a subject having AML a therapeutically effective amount of a conjugate of claim 2 .

18. The compound of claim 1 , wherein the IQB compound has the structure of Formula I.

19. The compound of claim 18 , wherein the L is a linker comprising the structure —NH-PEG 8 -C(O)-VAL-ALA-.

20. The compound of claim 19 , wherein L is a linker comprising the structure:

21. The compound of claim 1 , wherein the compound of Formula I has the structure:

22. The compound of claim 11 , wherein the anti-CLL-1 antibody has a light variable region comprising SEQ ID NO: 1 and a heavy variable region comprising SEQ ID NO:2.

23. The method of claim 17 , wherein the antibody-drug conjugate has the structure of Formula III:

wherein:

is an antibody or antibody fragment that binds to a cancer marker on a AML cell;

W—R M is a linking moiety formed by W and R x , wherein W is a moiety attached a natural or unnatural amino acid residue of the antibody/antibody fragment and R x is a reactive moiety linking L-IQB to the antibody, wherein R x is a succinimidyl, maleimidyl, cylooctynyl, aminooxy, bisulfonyl, sulfonyl, or isothiocyanate moiety, such that W—R M is a disulfide, a thiolated succinimidyl, an amino substituted succinimidyl, a (cyclooctyl)-1, 4 triazolyl, oxime substituted N-glycan, oxime, a substituted bis-sulfopropyl, a sulfonamidyl, an amide, or a thiocarbamate moiety;

L is a linker comprising the structure —NH-PEG 8 -C(O)-VAL-ALA-;

j is a number of 1 to 10; and,

IQB is a compound having a structure of Formula (I) or (II):

wherein:

the dotted bond shown between —C(R a )— and —N(R b )— or —C(R a′ )— and —N(R b′ )— is independently a single bond or a double bond;

each of R a and R a′ is independently H or OH;

each of R b and R b′ is not present, or is independently H, or a bond linked to linker L;

R 2 , R 2′ , R 3 , R 3′ , R 4 , R 4′ , R 6′ and R 6 are each independently selected from H, OH, C 1 -C 10 alkyl, C 2 -C 10 alkenyl or C 2 -C 10 alkynyl;

each of R 5 or R 5′ is independently NH 2 , CO 2 H, H, OH C 1 -C 10 alkyl, C 2 -C 10 alkenyl, C 2 -C 10 alkynyl, or a bond linked to linker L;

each of R 7 and R 7′ is H;

R 8 is:

H, NH 2 , CO 2 H, or a bond linked to linker L, wherein the carbon to which R 8 is attached also has a hydrogen substituent; or

an exo olefin having the structure

wherein the carbon to which R 8 is attached has no other substituent;

X is methylene;

each of Y and Y′ is independently O, S, or NH;

each of Z and Z′ is independently H, R, OH, OR, SH, SR, NH 2 , or NHR, where each R is independently unsubstituted C 1 -C 12 alkyl; and

wherein only one of R b , R b′ , R 5 , R 5′ , R 8 is a bond linked to linker L; and

wherein said IQB compound is enantiomerically pure.

24. The method of claim 23 , wherein the IQB compound has the structure of Formula I.

25. The method of claim 24 , wherein L is a linker comprising the structure:

26. The method of claim 17 , wherein the compound of Formula I has the structure:

27. The method of claim 17 , wherein the anti-CLL-1 antibody has a light variable region comprising SEQ ID NO: 1, and a heavy variable region comprising SEQ ID NO:2.

28. The method of claim 27 , wherein the —W—R M -L-IQB is a moiety having a structure of Formula IV:

29. A method of treating acute myeloid leukemia (AML) comprising administering to a subject having AML a therapeutically effective amount of a compound of claim 1 .

30. The method of claim 29 , wherein the IQB compound has the structure of Formula I.

31. The method of claim 29 , wherein: the IQB compound has the structure of Formula II.

32. The method of claim 30 , wherein the IQB compound has the structure of Formula I, wherein,

each of R a and R a′ is independently H, or OH;

if present, each of R b and R b′ is H;

R 2 , R 2′ , R 3 , R 3′ , R 4 , R 4′ , R 5 , R 5′ , R 6′ , R 6 , R 7 and R 7′ , are each H;

X is C 1-12 alkylene;

each of Y and Y′ is O; and

each of Z and Z′ is independently OR, where each R is independently unsubstituted C 1 -C 3 alkyl.

33. The method of claim 29 , wherein: the IQB compound of Formula I has the following structure: