Didemnin analogs and fragments and methods of making and using them

1. A method of performing a function selected from the group consisting of

I) inhibiting protein synthesis in a cell,

II) inhibiting growth of a cell,

III) inhibiting proliferation of a cell,

IV) inhibiting tumorigenesis in a cell, and

V) enhancing apoptosis of a cell,

the method comprising administering to the cell a didemnin analog having the structure

wherein:

i) R 1 is selected from the group consisting of

—H,

-(tert-butyloxycarbonyl),

-leucine,

—(N-methyl)leucine,

—(N-methyl)leucine-proline,

—(N-methyl)leucine-proline-lactate,

—(N-methyl)leucine-proline-pyruvate,

—(N-methyl)leucine-proline-lactate-(a first fluorophore),

—(N-methyl)leucine-proline-lactate-glutamine-pyroglutamate,

—(N-methyl)leucine-proline-lactate-glutamine-cyclopentanoate,

—(N-methyl)leucine-proline-alanine-leucine-pyroglutamate, and

—(N-methyl)leucine-proline-(N-methyl-alanine)-leucine-pyroglutamate;

ii) R 2 and R 3 are one of

(a) R 2 is selected from the group consisting of an isoleucine side chain, a valine side chain, an alanine side chain, a norleucine side chain, a norvaline side chain, a leucine side chain, a histidine side chain, a tryptophan side chain, an arginine side chain, a lysine side chain, a second fluorophore, and a substituent having the structure

and R 3 is selected from the group consisting of —CH 3 and —H; or

(b) R 2 and R 3 together form the structure

iii) each of R 5 , R 6 , R 7 , R 8 , and R 9 , when present, is independently selected from the group consisting of —H, —OH, —OCH 3 , —CO(C 6 H 5 ), —Br, —I, —F, —Cl, —CH 3 , and —C 2 H 5 ;

iv) R 4 is selected from the group consisting of an isoleucine side chain and a valine side chain;

v) X is selected from the group consisting of —O— and —NH—;

vi) Y is selected from the group consisting of —H and a hydroxyl protecting group;

vii) R 10 is selected from the group consisting of a leucine side chain and a lysine side chain; and

viii) the molecule is not tamandarin A and is not tamandarin B (compound 105).

2. The method of claim 1 , wherein R 1 is selected from the group consisting of

—H,

-(tert-butyloxycarbonyl),

-leucine,

—(N-methyl)leucine,

—(N-methyl)leucine-(S)proline,

—(N-methyl)leucine-(S)proline-(S)lactate,

—(N-methyl)leucine-(S)proline-pyruvate,

—(N-methyl)leucine-(S)proline-(S)lactate-(a first fluorophore),

—(N-methyl)leucine-(S)proline-(S)lactate-glutamine-pyroglutamate,

—(N-methyl)leucine-(S)proline-(S)lactate-glutamine-cyclopentanoate,

—(N-methyl)leucine-(S)proline-alanine-leucine-pyroglutamate, and

—(N-methyl)leucine-(S)proline-(N-methyl-alanine)-leucine-pyroglutamate.

3. The method of claim 1 , wherein R 1 is selected from the group consisting of

—H,

-(tert-butyloxycarbonyl),

-leucine,

—(N-methyl)leucine,

—(N-methyl)leucine-(S)proline,

—(N-methyl)leucine-(S)proline-(S)lactate,

—(N-methyl)leucine-(S)proline-pyruvate,

—(N-methyl)leucine-(S)proline-(S)lactate-(a first fluorophore),

—(N-methyl)leucine-(S)proline-(S)lactate-(S)glutamine-(S)pyroglutamate,

—(N-methyl)leucine-(S)proline-(S)lactate-(S)glutamine-(S)cyclopentanoate,

—(N-methyl)leucine-(S)proline-(S)alanine-(S)leucine-(S)pyroglutamate, and

—(N-methyl)leucine-(S)proline-(N-methyl-S-alanine)-(S)leucine-(S)pyroglutamate.

4. The method of claim 1 , wherein R 1 is —(N-methyl-R-leucine).

5. The method of claim 1 , wherein R 1 is —(N-methyl)leucine-(S)proline-pyruvate.

6. The method of claim 1 , wherein the didemnin analog is selected from the group consisting of

7. The method of claim 1 , wherein Y is —H, and wherein R 2 has the structure

8. The method of claim 1 , wherein R 2 is a lysine side chain and Y is —H.

9. The method of claim 1 , wherein the didemnin analog is selected from the group consisting of

10. The method of claim 1 , wherein the didemnin analog is administered as a composition further comprising a pharmaceutically acceptable carrier.

11. The method of claim 1 , wherein Y is —H, X is —O—, and R 1 is —(N-methyl-R-leucine)-(S)proline-(S)glutamine-(S)pyroglutamate.

12. The method of claim 11 , wherein the didemnin analog is selected from the group consisting of

13. The method of claim 1 , wherein R 4 is a valine side chain, Y is —H, and X is —O—.

14. The method of claim 13 , wherein the didemnin analog is selected from the group consisting of

15. The method of claim 1 , wherein R is —(N-methyl)leucine-(S)proline-(S)lactate-(a first fluorophore).

16. The method of claim 15 , wherein the didemnin analog is selected from the group consisting of

17. The method of claim 1 , wherein R 2 is a second fluorophore and wherein Y is —H.

18. The method of claim 17 , wherein the didemnin analog has the following structure, wherein FL is a fluorophore

19. The method of claim 1 , wherein X is —NH—.

20. The method of claim 19 , wherein the didemnin analog is selected from the group consisting of

21. A method of performing a function selected from the group consisting of

I) inhibiting protein synthesis in a cell,

II) inhibiting growth of a cell,

III) inhibiting proliferation of a cell,

IV) inhibiting tumorigenesis in a cell, and

V) enhancing apoptosis of a cell,

the method comprising administering to the cell a composition comprising a didemnin fragment having the structure

wherein:

i) Y is selected from the group consisting of —H and a hydroxyl protecting group;

ii) X is selected from the group consisting of —O— and —NH—;

iii) R 4 is selected from the group consisting of an isoleucine side chain and a valine side chain;

iv) APG is an amine protecting group; and

v) R 11 is selected from the group consisting of —OH, —NH 2 , —O(allyl), —O(pentafluorophenyl), and a substituent having the structure

wherein:

a) R 1 is selected from the group consisting of —H, and an amine protecting group;

b) R 2 and R 3 are one of

(I) R 2 is selected from the group consisting of an isoleucine side chain, a valine side chain, an alanine side chain, a norleucine side chain, a norvaline side chain, a leucine side chain, a histidine side chain, a tryptophan side chain, an arginine side chain, a lysine side chain, a second fluorophore, and a substituent having the structure

and R 3 is selected from the group consisting of —CH 3 and —H; or

(II) R 2 and R 3 together form having the structure

c) each of R 5 , R 6 , R 7 , R 8 , and R 9 , when present, is independently selected from the group consisting of —H, —OH, —OCH 3 , —CO(C 6 H 5 ), —Br, —I, —F, —Cl, —CH 3 , and —C 2 H 5 ;

d) R 10 is selected from the group consisting of a leucine side chain, a lysine side chain, and a protected lysine side chain; and

e) R 12 is selected from the group consisting of —H, and -2-(trimethylsilyl).

22. A method of performing a function selected from the group consisting of

I) inhibiting protein synthesis in a cell,

II) inhibiting growth of a cell,

III) inhibiting proliferation of a cell,

IV) inhibiting tumorigenesis in a cell, and

V) enhancing apoptosis of a cell,

the method comprising administering to the cell a didemnin analog having the structure

wherein:

i) R 2 and R 3 are one of

(a) R 3 is selected from the group consisting of —CH 3 and —H;

R 2 is selected from the group consisting of an isoleucine side chain, a valine side chain, an alanine side chain, a norleucine side chain, a norvaline side chain, a leucine side chain, a histidine side chain, a tryptophan side chain, an arginine side chain, a lysine side chain, a second fluorophore, and a substituent having the structure

and R 3 selected from the group consisting of —CH 3 and —H; or

(b) R 2 and R 3 together from the structure

ii) each of R 5 , R 6 , R 7 , R 8 , and R 9 , when present, is independently selected from the group consisting of —H, —OH, —OCH 3 , —CO(C 6 H 5 ), —Br, —I, —F, —Cl, —CH 3 , and —C 2 H 5 ;

iii) R 4 is selected from the group consisting of an isoleucine side chain and a valine side chain;

iv) X is selected from the group consisting of —O— and —NH—;

v) Y is selected from the group consisting of —H and a hydroxyl protecting group;

vi) R 10 is selected from the group consisting of a leucine side chain and a lysine side chain; and

vii) R 13 is an enzyme-cleavable moiety that is cleavable by an enzyme selected from the group consisting of a carboxypeptidase, a beta-lactamase, a beta galactosidase, a penicillin V-amidase, a cytosine deaminase, a nitroreductase, an alkaline phosphatase, a beta-glucuronidase, and a catalytic antibody.

23. The method of claim 22 , wherein R 13 has the structure

24. The method of claim 22 , wherein R 13 has the structure

25. The method of claim 22 , wherein the didemnin analog is selected from the group consisting of