Deoxo-proline-containing tamandarin and didemnin analogs, dehydro-proline-containing tamandarin and didemnin analogs, and methods of making and using them

1. A tamandarin compound having the structure

wherein:

i) R 1 is selected from the group consisting of

—(N-methyl)leucine-deoxo-proline,

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

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

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

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

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

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

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

—(N-methyl)leucine-dehydro-proline,

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

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

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

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

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

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

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

ii) R 2 and R 3 are one of

(a) R 3 is selected from the group consisting of —CH3 and —H; and 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

(b) R 2 and R 3 together are a substituent having 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 —C2H 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; and

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

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

—(N-methyl)leucine-deoxo-(S)proline,

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

4. The compound of claim 1 , wherein R 2 is

R 3 is methyl, R 4 is an isoleucine side chain, each of R 5 , R 6 , R 8 , and R 9 is —H, R 7 is methoxy, R 10 is a leucine side chain, X is —O—, and Y is —H.

5. The compound of claim 1 , wherein the tamandarin is compound 201 having the structure

6. The compound of claim 1 , wherein the tamandarin is compound 203 having the structure

7. The compound of claim 1 , wherein R 1 is

—(N-methyl)leucine-deoxo-(S)proline-lactate.

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

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

10. The compound of claim 1 , wherein the tamandarin has the following structure, wherein FL is a fluorophore

11. The compound of claim 1 , wherein X is —NH—.

12. A composition comprising the compound of claim 1 and a pharmaceutically acceptable carrier.

13. A support having the tamandarin compound of claim 1 covalently attached thereto.

14. A method of inhibiting protein synthesis in a cell, the method comprising administering the compound of claim 1 to the cell.

15. A method of inhibiting growth of a cell, the method comprising administering the compound of claim 1 to the cell.

16. A method of inhibiting proliferation of a cell, the method comprising administering the compound of claim 1 to the cell.

17. A method of inhibiting tumorigenesis in a cell, the method comprising administering the compound of claim 1 to the cell.

18. A method of enhancing apoptosis of a cell, the method comprising administering the compound of claim 1 to the cell.

19. A compound having a structure selected from the group consisting of

wherein:

i) R 2 and R 3 are one of

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

(b) R 2 and R 3 together are a substituent having 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.

20. The compound of claim 19 , wherein R 13 has the structure

21. The compound of claim 19 , wherein R 13 has the structure

22. A method of inhibiting protein synthesis in a cell, the method comprising administering the compound of claim 19 to the cell.

23. A method of inhibiting growth of a cell, the method comprising administering the compound of claim 19 to the cell.

24. A method of inhibiting proliferation of a cell, the method comprising administering the compound of claim 19 to the cell.

25. A method of inhibiting tumorigenesis in a cell, the method comprising administering the compound of claim 19 to the cell.

26. A method of enhancing apoptosis of a cell, the method comprising administering the compound of claim 19 to the cell.

27. A didemnin compound having the structure

wherein:

i) R 1 is selected from the group consisting of

—(N-methyl)leucine-deoxo-proline,

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

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

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

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

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

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

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

—(N-methyl)leucine-dehydro-proline,

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

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

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

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

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

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

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

ii) R 2 and R 3 are one of

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

(b) R 2 and R 3 together are a substituent having 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; and

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

28. The compound of claim 27 , wherein R 1 is selected from the group consisting of

—(N-methyl)leucine-deoxo-(S)proline,

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

29. The compound of claim 27 , wherein R 1 is selected from the group consisting of

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

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

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

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

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

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

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

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

30. The compound of claim 27 , wherein R 2 is

R 3 is methyl, R 4 is an isoleucine side chain, each of R 5 , R 6 , R 8 , and R 9 is —H, R 7 is methoxy, R 10 is a leucine side chain, X is —O—, and Y is —H.

31. The compound of claim 27 , wherein the didemnin is compound 202 having the structure

32. The compound of claim 27 , wherein the didemnin is compound 204 having the structure

33. The compound of claim 27 , wherein R 1 is

—(N-methyl)leucine-deoxo-(S)proline-lactate.

34. The compound of claim 27 , wherein Y is —H, and wherein R 2 has the structure

35. The compound of claim 27 , wherein R 2 is a lysine side chain and Y is —H.

36. The compound of claim 27 , wherein X is —NH—.

37. A composition comprising the compound of claim 27 and a pharmaceutically acceptable carrier.

38. A support covalently attached with the didemnin compound of claim 27 .

39. A method of inhibiting protein synthesis in a cell, the method comprising administering the compound of claim 27 to the cell.

40. A method of inhibiting growth of a cell, the method comprising administering the compound of claim 27 to the cell.

41. A method of inhibiting proliferation of a cell, the method comprising administering the compound of claim 27 to the cell.

42. A method of inhibiting tumorigenesis in a cell, the method comprising administering the compound of claim 27 to the cell.

43. A method of enhancing apoptosis of a cell, the method comprising administering the compound of claim 27 to the cell.

44. A method of preparing a tamandarin or didemnin compound comprising incorporating a deoxo-proline residue in place of a proline residue of the compound in a chemical reaction to prepare said tamandarin or didemnin analog.

45. The method of claim 44 , wherein the compound comprises an (N-methyl)leucine-proline moiety and wherein the (N-methyl)leucine-proline moiety is replaced by an (N-methyl)leucine-deoxo-proline moiety.

46. The method of claim 45 wherein the (N-methyl)leucine-deoxo-proline is made by

reducing the ester function of proline to an aldehyde function; and

coupling the proline with the (N-methyl)leucine moiety by reductive amination to yield the (N-methyl)leucine-deoxo-proline moiety.

47. The method of claim 46 , wherein the amine moiety of the proline is protected with an amine-protecting group prior to the reductive amination.

48. The method of claim 46 , wherein the ester function of the proline is reduced to an aldehyde function by contacting the proline with a strong base and then contacting the proline with an oxidizing agent.

49. The method of claim 46 , wherein the reductive amination is performed in a non-aqueous solvent in the presence of a strong base and a carboxylic acid catalyst.

50. A method of preparing a tamandarin or didemnin compound the improvement comprising incorporating a dehydro-proline residue in place of a proline residue of the compound in a chemical reaction used to prepare said tamandarin or didemnin compound.

51. The method of claim 50 , wherein the compound comprises an (N-methyl)leucine-proline moiety and wherein the (N-methyl)leucine-proline moiety is replaced by an (N-methyl)leucine-dehydro-proline moiety.

52. The method of claim 50 , wherein the dehydro-proline residue is made by protecting the carboxyl and amino moieties of the 4-hydroxyprolinate, alkyl sulfonylating the 4-hydroxyl moiety, displacing the alkyl-sulfonate moiety with an aryl-selenyl moiety, oxidatively eliminating the aryl-selenyl moiety to yield a dehydro-proline moiety having protected carboxyl and amine moieties, and coupling the dehydro-proline moiety with an amine moiety of the analog.

53. The method of claim 50 , wherein the alkyl-sulfonate moiety is a methyl-sulfonate moiety.

54. The method of claim 50 , wherein the aryl-selenyl moiety is a phenyl-selenyl moiety.

55. The method of claim 50 , wherein the 4-hydroxyprolinate is trans-4-hydroxyprolinate.

56. The compound of claim 1 , wherein the compound is substantially pure.

57. The compound of claim 5 , wherein the compound is substantially pure.

58. The compound of claim 6 , wherein the compound is substantially pure.

59. The compound of claim 10 , wherein the compound is substantially pure.

60. The compound of claim 19 , wherein the compound is substantially pure.

61. The compound of claim 27 , wherein the compound is substantially pure.

62. A composition comprising the compound of claim 19 and a pharmaceutically acceptable carrier.