Derivatives of uncialamycin, methods of synthesis and their use as antitumor agents

1. A compound of the formula:

wherein:

Y 1 is —O(CH 2 ) m Y′, —NH(CH 2 ) m Y′, —S(CH 2 ) m Y′, or —(CH 2 ) m NR 1 R 2 , or is taken together with Z 1 as defined below;

wherein:

Y′ is hydroxy, halo, mercapto, alkyl (C1-12) , substituted alkyl (C1-12) , alkenyl (C2-12) , substituted alkenyl (C2-12) , alkynyl (C2-12) , substituted alkynyl (C2-12) , acyl (C≦12) , substituted acyl (C1-12) , acyloxy (C1-12) , substituted acyloxy (C1-12) , alkylamino (C1-12) , or substituted alkylamino (C1-12) ;

m is 1, 2, 3, 4, 5, or 6; and

R 1 and R 2 are each independently selected from hydrogen, hydroxy, alkyl (C1-12) , substituted alkyl (C1-12) , alkenyl (C2-12) , substituted alkenyl (C2-12) , alkynyl (C2-12) , substituted alkynyl (C2-12) , aryl (C6-12) , substituted aryl (C6-12) , aralkyl (C7-12) , substituted aralkyl (C7-12) , heteroaryl (C1-12) , substituted heteroaryl (C1-12) , heterocycloalkyl (C2-12) , substituted heterocycloalkyl (C2-12) , acyl (C1-12) , substituted acyl (C1-12) , acyloxy (C1-12) , substituted acyloxy (C1-12) , alkylamino (C1-12) , substituted alkylamino (C1-12) ; a monovalent amine protecting group, —C(O)O(CH 2 ) n S-A 1 , —C(O)O(CH 2 ) n S(O)-A 1 , or —C(O)O(CH 2 ) n S(O) 2 -A 1 ,

wherein:

A 1 is aryl (C6-12) , substituted aryl (C6-12) , or

wherein:

A 2 is alkenyl (C2-12) , substituted alkenyl (C2-12) , alkynyl (C2-12) , substituted alkynyl (C2-12) , acyl (C1-12) , substituted acyl (C1-12) , acyloxy (C1-12) , substituted acyloxy (C1-12) , alkoxy (C1-12) , substituted alkoxy (C1-12) , alkylamino (C1-12) , substituted alkylamino (C1-12) , dialkylamino (C2-12) , or substituted dialkylamino (C2-12) ; wherein A 2 is not —CO 2 H, —CO 2 CH 3 , —OCH 3 , —OCH 2 CH 3 , —C(O)CH 3 , —NHCH 3 , —NHCH 2 CH 3 , —N(CH 3 ) 2 , —C(O)NH 2 , and —OC(O)CH 3 ; and

n is 1, 2, 3, 4, or 5; or

R 1 and R 2 are taken together and are divalent amine protecting group, alkanediyl (C1-12) , alkylaminodiyl (C1-8) ; alkoxydiyl (C1-8) ; or a substituted version of either of these groups; or

Y 1 is taken with Z 1 and is alkylaminodiyl (C1-8) substituted alkylaminodiyl (C1-8) ; -alkanediyl (C1-6) -NZ 2 -alkanediyl (C1-6) , or -substituted alkanediyl (C1-6) -NZ 2 -substituted alkanediyl (C1-6) ,

wherein:

Z 2 is hydrogen, an amine protecting group, acyl (C6-12) , substituted acyl (C6-12) , —C(O)O(CH 2 ) n S-A 3 , or —C(O)O(CH 2 ) n S(O) 2 -A 3 , and

wherein:

A 3 is aryl (C6-12) , substituted aryl (C6-12) , or

wherein:

 A 4 is alkenyl (C2-12) , substituted alkenyl (C2-12) , alkynyl (C2-12) , substituted alkynyl (C2-12) , acyl (C1-12) , substituted acyl (C1-12) , acyloxy (C1-12) , substituted acyloxy (C1-12) , alkoxy (C1-12) , substituted alkoxy (C1-12) , alkylamino (C1-12) , substituted alkylamino (C1-12) , dialkylamino (C2-12) , or substituted dialkylamino (C2-12) , wherein A 4 is not —CO 2 H, —CO 2 CH 3 , —OCH 3 , —OCH 2 CH 3 , —C(O)CH 3 , —NHCH 3 , —NHCH 2 CH 3 , —N(CH 3 ) 2 , —C(O)NH 2 , and —OC(O)CH 3 ;

Z 1 is absent, hydrogen or taken together with Y 1 as defined above;

R 3 and Z 2 are each independently selected from hydrogen, hydroxy, halo, amino, cyano, nitro, phosphate, or mercapto, or

alkyl (C1-12) , alkenyl (C2-12) , alkynyl (C2-12) , aryl (C6-12) , aralkyl (C7-12) , heteroaryl (C1-12) , heterocycloalkyl (C2-12) , acyl (C1-12) , alkoxy (C1-12) , acyloxy (C1-12) , alkylamino (C1-12) , dialkylamino (C2-12) , amido (C1-12) , or a substituted version of any of these groups;

o is 1, 2, or 3;

R 4 is hydrogen, alkyl (C1-12) , a monovalent amine protecting group, or substituted alkyl (C1-12) ;

R 5 , R 6 , and R 7 are each independently hydrogen, hydroxy, amino, mercapto, —OX 1 , —NX 2 X 3 , or —SX 4 ; or

alkyl (C1-12) , alkoxy (C1-12) , acyloxy (C1-12) , alkylamino (C2-12) , alkylthio (C1-12) , amido (C1-12) , or a substituted version of any of these groups;

wherein:

X 1 is a hydroxy protecting group;

X 2 and X 3 are independently selected from hydrogen, a monovalent amine protecting group, or when X 2 and X 3 are taken together form a divalent amine protecting group; and

X 4 is a thiol protecting group;

R 8 is hydroxy, amino, or mercapto; or

alkoxy (C1-12) , acyloxy (C1-12) , alkylamino (C1-12) , dialkylamino (C2-12) , alkylthio (C1-12) , amido (C1-12) , or a substituted version of any of these groups; and

R 9 , R 10 , R 11 , and R 12 are each independently selected from hydrogen, hydroxy, amino, mercapto, —OX 1 , —NX 2 X 3 , or —SX 4 , or

alkyl (C1-12) , alkenyl (C2-12) , alkynyl (C2-12) , aryl (C6-12) , aralkyl (C7-12) , heteroaryl (C1-12) , heterocycloalkyl (C2-12) , acyl (C1-12) , alkoxy (C1-12) , acyloxy (C1-12) , alkylamino (C1-12) , dialkylamino (C2-12) , alkylthio (C1-12) , amido (C1-12) , or a substituted version of any of these groups; or Y 2 —R 13 ;

wherein:

X 1 is alkyl (C1-12) , substituted alkyl (C1-12) , alkenyl (C2-12) , substituted alkenyl (C2-12) , alkynyl (C2-12) , substituted alkynyl (C2-12) , aryl (C6-12) , substituted aryl (C6-12) , aralkyl (C7-12) , substituted aralkyl (C7-12) , heteroaryl (C1-12) , substituted heteroaryl (C1-12) , heterocycloalkyl (2-12) , substituted heterocycloalkyl (C2-12) , acyl (C1-12) , substituted acyl (C1-12) , or a hydroxy protecting group;

X 2 and X 3 are independently selected from hydrogen, alkyl (C1-12) , substituted alkyl (C1-12) , alkenyl (C2-12) , substituted alkenyl (C2-12) , alkynyl (C2-12) , substituted alkynyl (C2-12) , aryl (C6-12) , substituted aryl (C6-12) , aralkyl (C7-12) , substituted aralkyl (C7-12) , heteroaryl (C1-12) , substituted heteroaryl (C1-12) , heterocycloalkyl (C2-12) , substituted heterocycloalkyl (C2-12) , acyl (C1-12) , substituted acyl (C1-12) , a monovalent amine protecting group, or when X 2 and X 3 are taken together form a divalent amine protecting group;

X 4 is alkyl (C1-12) , substituted alkyl (C1-12) , alkenyl (C2-12) , substituted alkenyl (C2-12) , alkynyl (C2-12) , substituted alkynyl (C2-12) , aryl (C6-12) , substituted aryl (C6-12) , aralkyl (C7-12) , substituted aralkyl (C7-12) , heteroaryl (C1-12) , substituted heteroaryl (C1-12) , heterocycloalkyl (C2-12) , substituted heterocycloalkyl (C2-12) , acyl (C1-12) , substituted acyl (C1-12) , or a thiol protecting group;

Y 2 is alkanediyl (C1-12) or substituted alkanediyl (C1-12) ; and

R 13 is hydroxy, amino, mercapto, —OX 1 , —NX 2 X 3 , or —SX 4 , or heteroaryl (C1-12) , heterocycloalkyl (C2-12) , acyl (C1-12) , alkoxy (C1-12) , acyloxy (C1-12) , alkylamino (C1-12) , dialkylamino (C2-12) , alkylthio (C1-12) , amido (C1-12) , or a substituted version of any of these groups;

p and q are each independently 1 or 2; and

Z 3 is hydrogen, hydroxy, halo, amino, cyano, nitro, phosphate, or mercapto, or

alkyl (C1-12) , alkenyl (C2-12) , alkynyl (C2-12) , aryl (C6-12) , aralkyl (C7-12) , heteroaryl (C1-12) , heterocycloalkyl (C2-12) , acyl (C1-12) , alkoxy (C1-12) , acyloxy (C1-12) , alkylamino (C1-12) , dialkylamino (C2-12) , amido (C1-12) , or a substituted version of any of these groups;

provided that Y 1 is not —NHMe or —NHCH 2 CH 2 NH 2 ; or

a pharmaceutically acceptable salt thereof.

2. The compound of claim 1 , wherein the formula is further defined as:

wherein:

Y 1 is —O(CH 2 ) m Y′, —NH(CH 2 ) m Y′, —S(CH 2 ) m Y′, or —(CH 2 ) m NR 1 R 2 , or taken together with Z 1 as described below;

wherein:

Y′ is hydroxy, halo, mercapto, alkenyl (C2-12) , substituted alkenyl (C2-12) , alkynyl (C2-12) , substituted alkynyl (C2-12) , acyl (C≦12) , substituted acyl (C1-12) , acyloxy (C1-12) , substituted acyloxy (C1-12) , alkylamino (C1-12) , or substituted alkylamino (C1-12) ;

m is 1, 2, 3, 4, 5, or 6; and

R 1 and R 2 are each independently selected from hydrogen, hydroxy, alkyl (C1-12) , substituted alkyl (C1-12) , alkenyl (C2-12) , substituted alkenyl (C2-12) , alkynyl (C2-12) , substituted alkynyl (C2-12) , aryl (C6-12) , substituted aryl (C6-12) , aralkyl (C7-12) , substituted aralkyl (C7-12) , heteroaryl (C1-12) , substituted heteroaryl (C1-12) , heterocycloalkyl (C2-12) , substituted heterocycloalkyl (C2-12) , acyl (C1-12) , substituted acyl (C1-12) , acyloxy (C1-12) , substituted acyloxy (C1-12) , alkylamino (C1-12) , substituted alkylamino (C1-12) ; a monovalent amine protecting group, —C(O)O(CH 2 ) n S-A 1 , —C(O)O(CH 2 ) n S(O)-A 1 , or —C(O)O(CH 2 ) n S(O) 2 -A 1 ,

wherein:

A 1 is aryl (C6-12) or substituted aryl (C6-12) ; and

n is 1, 2, 3, 4, or 5;

R 1 and R 2 are taken together and are a divalent protecting group, alkanediyl (C≦12) , alkylaminodiyl (C≦8) ; alkoxydiyl (C≦8) ; or a substituted version of either of these groups; or

Y 1 is taken with Z 1 and is alkylaminodiyl (C1-8) substituted alkylaminodiyl (C1-8) ; or -alkanediyl (C1-6) -NZ 2 -alkanediyl (C1-6) ,

wherein:

Z 2 is hydrogen, an amine protecting group, acyl (C6-12) , substituted acyl (C6-12) , —C(O)O(CH 2 ) n S-A 3 , or —C(O)O(CH 2 ) n S(O) 2 -A 3 ,

wherein:

A 3 is aryl (C6-12) or substituted aryl (C6-12) ;

Z 1 is absent, hydrogen or taken together with Y 1 as defined above;

R 3 is hydrogen, hydroxy, halo, amino, cyano, nitro, phosphate, or mercapto, or

alkyl (C1-12) , alkenyl (C2-12) , alkynyl (C2-12) , aryl (C6-12) , aralkyl (C7-12) , heteroaryl (C1-12) , heterocycloalkyl (C2-12) , acyl (C1-12) , alkoxy (C1-12) , acyloxy (C1-12) , alkylamino (C1-12) , dialkylamino (C2-12) , amido (C1-12) , or a substituted version of any of these groups;

o is 1, 2, or 3;

R 4 is hydrogen, alkyl (C1-12) , a monovalent amine protecting group, or substituted alkyl (C1-12) ;

R 5 , R 6 , and R 7 are each independently hydrogen, hydroxy, amino, mercapto, —OX 1 , —NX 2 X 3 , or —SX 4 ; or

alkyl (C1-12) , alkoxy (C1-12) , acyloxy (C1-12) , alkylamino (C1-12) , dialkylamino (C2-12) , alkylthio (C1-12) , amido (C1-12) , or a substituted version of any of these groups;

wherein:

X 1 is a hydroxy protecting group;

X 2 and X 3 are independently selected from hydrogen, a monovalent amine protecting group, or when X 2 and X 3 are taken together form a divalent amine protecting group; and

X 4 is a thiol protecting group; and

R 8 is hydroxy, amino, or mercapto; or

alkoxy (C1-12) , acyloxy (C1-12) , alkylamino (C1-12) , dialkylamino (C2-12) , alkylthio (C1-12) , amido (C1-12) , or a substituted version of any of these groups; or

a pharmaceutically acceptable salt thereof.

3. The compound of claim 1 , wherein the formula is further defined as:

wherein:

R 1 and R 2 are each independently selected from hydrogen, hydroxy,

alkyl (C1-12) , substituted alkyl (C1-12) , alkenyl (C2-12) , substituted alkenyl (C2-12) , alkynyl (C2-12) , substituted alkynyl (C2-12) , aryl (C6-12) , substituted aryl (C6-12) , aralkyl (C7-12) , substituted aralkyl (C7-12) , heteroaryl (C1-12) , substituted heteroaryl (C1-12) , heterocycloalkyl (C2-12) , substituted heterocycloalkyl (C2-12) , acyl (C1-12) , substituted acyl (C1-12) , acyloxy (C1-12) , substituted acyloxy (C1-12) , alkylamino (C1-12) , substituted alkylamino (C1-12) ; a monovalent amine protecting group, —C(O)O(CH 2 ) n S-A 1 , —C(O)O(CH 2 ) n S(O)-A 1 , or —C(O)O(CH 2 ) n S(O) 2 -A 1 ,

wherein:

A 1 is aryl (C6-12) or substituted aryl (C6-12) ; and

n is 1, 2, 3, 4, or 5;

R 3 is hydrogen, hydroxy, halo, or alkoxy (C1-12) or substituted alkoxy (C1-12) ;

o is 1, 2, or 3;

R 4 is hydrogen, a monovalent amine protecting group, alkyl (C1-12) , or substituted alkyl (C1-12) ;

R 5 , R 6 , and R 7 are each independently hydrogen, hydroxy, amino, mercapto, —OX 1 , —NX 2 X 3 , or —SX 4 ; or

alkyl (C1-12) or substituted alkyl (C1-12) ;

wherein:

X 1 is a hydroxy protecting group;

X 2 and X 3 are independently selected from hydrogen, a monovalent amine protecting group, or when X 2 and X 3 are taken together form a divalent amine protecting group; and

X 4 is a thiol protecting group; and

R 8 is hydroxy, amino, or mercapto; or

a pharmaceutically acceptable salt thereof.

4. The compound of claim 1 , wherein the formula is further defined as:

wherein:

R 1 and R 2 are each independently selected from hydrogen, hydroxy,

alkyl (C1-12) , substituted alkyl (C1-12) , aryl (6-12) , substituted aryl (C6-12) , aralkyl (C7-12) , substituted aralkyl (C7-12) , acyl (C1-12) , substituted acyl (C1-12) , a monovalent amine protecting group, —C(O)O(CH 2 ) n S—C 6 H 5 , —C(O)O(CH 2 ) n S(O)—C 6 H 5 , or —C(O)O(CH 2 ) n S(O) 2 —C 6 H 5 ,

a monovalent amine protecting group, or R 1 and R 2 are taken together and form a divalent amine protecting group, or

alkyl (C1-12) , aryl (C6-12) , aralkyl (C7-12) , acyl (C1-12) , or a substituted version of any of these groups;

R 3 is hydrogen, hydroxy, halo, or

alkoxy (C1-12) or substituted alkoxy (C1-12) ; and

o is 1, 2, or 3; or

a pharmaceutically acceptable salt thereof.

5. The compound of claim 1 , wherein the formula is further defined as:

wherein:

X 1 is a hydroxy protecting group;

R 1 and R 2 are each independently selected from hydrogen, alkyl (C1-12) , or substituted alkyl (C1-12) ; and

R 3 is hydrogen, alkoxy (C1-12) , or substituted alkoxy (C1-12) ; and

o is 2;

or a pharmaceutically acceptable salt thereof.

6. The compound of claim 1 , wherein Y 1 is —(CH 2 ) m NR 1 R 2 .

7. The compound of claim 1 , wherein m is 1, 2, or 3.

8. The compound of claim 1 , wherein R 1 or R 2 is hydrogen, alkyl (C1-12) or substituted alkyl (C1-12) .

9. The compound of claim 1 , wherein R 3 is hydrogen.

10. The compound of claim 1 , wherein the compound is further defined as:

or a pharmaceutically acceptable salt thereof.

11. The compound of claim 1 , further defined as:

or a pharmaceutically acceptable salt thereof.

12. A conjugate of the formula:

(A 5 -L) r -A 6   (XVII)

wherein:

A 5 is a compound of claim 1 ;

L is a linker;

r is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and

A 6 is a cell targeting moiety.

13. The conjugate of claim 12 , wherein L comprises a polypeptide cleavable by an intracellular enzyme.

14. The conjugate of claim 13 , wherein the enzyme is cathepsin B.

15. The conjugate of claim 12 , wherein L comprises a self-immolating group.

16. The conjugate of claim 12 , wherein A 6 is an antibody whose antigen is a tumor associated antigen.

17. The conjugate of claim 16 , wherein the antigen is mesothelin, glypican-3, or CD70 and the antibody is an anti-mesothelin, anti-glypican-3, or anti-CD70 antibody.

18. The conjugate of claim 12 , wherein A 5 is a compound according to formula I.

19. The conjugate of claim 12 , further comprising a structure of the formula:

wherein:

A 6 is an antibody and r is 1, 2, 3, or 4.

20. The compound of claim 11 further defined as:

or a pharmaceutically acceptable salt thereof.

21. The compound of claim 11 further defined as:

or a pharmaceutically acceptable salt thereof.