Texaphyrin metal complex mediated ester hydrolysis

• US 5,559,207 A
• Filed: 04/14/1994
• Issued: 09/24/1996
• Est. Priority Date: 03/06/1989
• Status: Expired due to Fees

• Current Assignee: Pharmacyclics Incorporated (Abbvie Incorporated), Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Pharmacyclics Incorporated (Abbvie Incorporated), Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Ross, Kevin L., Miller, Richard A., Sessler, Jonathan L., Magda, Darren, Iverson, Brent, Dow, William C., Wright, Meredith, Smith, Daniel A., Kr al, Vladimir A.
• Primary Examiner(s): Patterson, Jr., Charles L.
• Application Number:
  US08/227,370
• Time in Patent Office: 894 Days
• Field of Search: 530/387.1, 530/300, 536/23.1, 536/1.11, 534/11, 534/15, 534/16, 435/188.5, 435/91.1, 435/135, 435/155
• US Class Current: 530/300
• CPC Class Codes: A61K 41/0038 Radiosensitizing, i.e. administration of pharmaceutical agents that enhance the effect of radiotherapy radiotherapy per se A61N5/10
• CPC Class Codes: A61K 41/0076 PDT with expanded (metallo)porphyrins, i.e. having more than 20 ring atoms, e.g. texaphyrins, sapphyrins, hexaphyrins, pentaphyrins, porphocyanines
• CPC Class Codes: A61K 47/546 Porphyrines; Porphyrine with an expanded ring system, e.g. texaphyrine
• CPC Class Codes: A61K 49/0021 the fluorescent group being a small organic molecule
• CPC Class Codes: A61K 49/0036 Porphyrins used in photodynamic therapy A61K41/0071 or A61K41/0076; used as targeting group or modifying agent for targeting a therapeutic compound A61K47/546
• CPC Class Codes: A61K 49/085 conjugated systems
• CPC Class Codes: A61K 49/106 the complex-forming compound being cyclic, e.g. DOTA
• CPC Class Codes: A61K 49/126 Linear polymers, e.g. dextran, inulin, PEG
• CPC Class Codes: A61K 51/0485 Porphyrins, texaphyrins wherein the nitrogen atoms forming the central ring system complex the radioactive metal
• CPC Class Codes: A61L 2/0011 using physical methods
• CPC Class Codes: B01J 2531/025 Ligands with a porphyrin ring system or analogues thereof, e.g. phthalocyanines, corroles
• CPC Class Codes: B01J 2531/23 Calcium
• CPC Class Codes: B01J 2531/26 Zinc
• CPC Class Codes: B01J 2531/27 Cadmium
• CPC Class Codes: B01J 2531/28 Mercury
• CPC Class Codes: B01J 2531/38 Lanthanides other than lanthanum
• CPC Class Codes: B01J 2531/72 Manganese
• CPC Class Codes: B01J 2531/842 Iron
• CPC Class Codes: B01J 2531/845 Cobalt
• CPC Class Codes: B01J 2531/847 Nickel
• CPC Class Codes: B01J 2531/96 : Water
• CPC Class Codes: B01J 31/061 : Chiral polymers
• CPC Class Codes: B01J 31/1658 : immobilised by covalent linkages, i.e. pendant complexes with optional linking groups, e.g. on Wang or Merrifield resins
• CPC Class Codes: B01J 31/1815 : with more than one complexing nitrogen atom, e.g. bipyridyl, 2-aminopyridine
• CPC Class Codes: C07D 487/22 : in which the condensed system contains four or more hetero rings
• CPC Class Codes: C07H 21/00 : Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

A method of phosphate ester hydrolysis including incubating a solution of an aqueous phosphate ester with a texaphyrin metal complex. The metal is a metal cation having catalytic activity for ester bond hydrolysis in aqueous solution, in particular, a lanthanide metal cation, preferably Eu(III) or Dy(III). A preferred substrate is RNA and a preferred texaphyrin is a derivatized texaphyrin having binding specificity, in particular, a texaphyrin covalently coupled to a site-directed molecule, preferably an oligonucleotide.

Claims

1. A method of phosphate ester hydrolysis, comprising

obtaining an aqueous phosphate ester solution, and

incubating the solution with a texaphyrin metal complex, said incubation being under conditions and for a time sufficient to hydrolyze the phosphate ester.

2. The method of claim 1 where the metal is a divalent or a trivalent metal cation having catalytic activity for ester bond hydrolysis in aqueous solution.

3. The method of claim 1 where the metal is a lanthanide cation.

4. The method of claim 1 where the metal is La(III), Nd(III), Sm(III), Gd(III), Tm(III), or Lu(III).

5. The method of claim 1 where the metal is Eu(III) or Dy(III).

6. The method of claim 1 where the phosphate ester is RNA.

7. A method of phosphate ester hydrolysis, comprising incubating an aqueous phosphate ester solution with a texaphyrin metal complex, said incubation being under conditions and for a time sufficient to hydrolyze the phosphate ester, wherein the texaphyrin metal complex has the structure:

##STR3## where;

M is a divalent or a trivalent metal cation having catalytic activity for ester bond hydrolysis in aqueous solution;

R₁, R₂, R₃, R₄, R₅ and R₆ are independently hydrogen, hydroxyl, alkyl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, saccharide, carboxyalkyl, carboxyamidealkyl, a site-directed molecule or a couple to a site-directed molecule or to a catalytic group; and

N is an integer less than or equal to 2.

8. The method of claim 7 where at least one of R₁, R₂, R₃, R₄, R₅ and R₆ is a site-directed molecule or is a couple to a site-directed molecule.

9. The method of claim 7 where the site-directed molecule is an oligonucleotide, a hormone, an antibody, a texaphyrin molecule, a sapphyrin molecule, or a peptide having affinity for a biological receptor.

10. The method of claim 7 where the oxyhydroxyalkyl is C.sub.(n-x) H.sub.((2n+1)-2x) O_x O_y or OC.sub.(n-x) H.sub.((2n+1)-2x) O_x O_y where

n is a positive integer from 1 to 10;

x is zero or a positive integer less than or equal to n; and

y is zero or a positive integer less than or equal to ((2n+1)-2x).

11. The method of claim 7 where the oxyhydroxyalkyl or saccharide is C_n H.sub.((2n+1)-q) O_y R^a_q, OC_n H.sub.((2n++1)-q) O_y R^a_q or (CH₂)_n CO₂ R^a where

n is a positive integer from 1 to 10,

y is zero or a positive integer less than ((2n+1)-q),

q is zero or a positive integer less than or equal to 2n+1,

R^a is independently H, alkyl, hydroxyalkyl, saccharide, C.sub.(m-w) H.sub.((2m+1)-2w) O_w O_z, O₂ CC.sub.(m-w) H.sub.((2m+1)-2w) O_w O_z or N(R)OCC.sub.(m-w) H.sub.((2m+1)-2w) O_w O_z, where

m is a positive integer from 1 to 10,

w is zero or a positive integer less than or equal to m,

z is zero or a positive integer less than or equal to ((2m+1)-2w),

R is H, alkyl, hydroxyalkyl, or C_m H.sub.((2m+1)-r) O_z R^b_r where

m is a positive integer from 1 to 10,

z is zero or a positive integer less than ((2m+1)-r),

r is zero or a positive integer less than or equal to 2m+1, and

R^b is independently H, alkyl, hydroxyalkyl, or saccharide.

12. The method of claim 7 where the carboxyamidealkyl is (CH₂)_n CONHR^a, O(CH₂)_n CONHR^a, (CH₂)_n CON(R^a)₂, or O(CH₂)_n CON(R^a)₂ where

n is a positive integer from 1 to 10,

R^a is independently H, alkyl, hydroxyalkyl, saccharide, C.sub.(m-w) H.sub.((2m+1)-2w) O_w O_z, O₂ CC.sub.(m-w) H.sub.((2m+1)-2w) O_w O_z, N(R)OCC.sub.(m-w) H.sub.((2m+1)-2w) O_w O_z, or a site-directed molecule or a catalytic group where

m is a positive integer from 1 to 10,

w is zero or a positive integer less than or equal to m,

z is zero or a positive integer less than or equal to ((2m+1)-2w),

R is H, alkyl, hydroxyalkyl, or C_m H.sub.((2m+1)-r) O_z R^b_r where

m is a positive integer from 1 to 10,

z is zero or a positive integer less than ((2m+1)-r),

r is zero or a positive integer less than or equal to 2m+1, and

R^b is independently H, alkyl, hydroxyalkyl, or saccharide.

13. The method of claim 12 where R^a is an oligonucleotide.

14. The method of claim 7 where the carboxyalkyl is C_n H.sub.((2n+1)-q) O_y R^c_q or OC_n H.sub.((2n+1)-q) O_y R^c_q where

n is a positive integer from 1 to 10;

y is zero or a positive integer less than ((2n+1)-q),

q is zero or a positive integer less than or equal to 2n+1,

R^c is (CH₂)_n CO₂ R^d, (CH₂)_n CONHR^d, (CH₂)_n CON(R^d)₂ or a site-directed molecule or a catalytic group where

n is a positive integer from 1 to 10;

R^d is independently H, alkyl, hydroxyalkyl, saccharide, C.sub.(m-w) H.sub.((2m+1)-2w) O_w O_z, O₂ CC.sub.(m-w) H.sub.((2m+1)-2w) O_w O_z or N(R)OCC.sub.(m-w) H.sub.((2m+m)-2w) O_w O_z, where m is a positive integer from 1 to 10,

w is zero or a positive integer less than or equal to m,

z is zero or a positive integer less than or equal to ((2m+1)-2w),

R is H, alkyl, hydroxyalkyl, or C_m H.sub.((2m+1)-r) O_z R^b_r where

m is a positive integer from 1 to 10,

z is zero or a positive integer less than ((2m+1)-r),

r is zero or a positive integer less than or equal to 2m+1, and R^b is independently H, alkyl, hydroxyalkyl, or saccharide.

15. The method of claim 14 where R^c is an oligonucleotide.

16. The method of claim 7 wherein the phosphate ester is RNA.

17. The method of claim 7 wherein M is a lanthanide cation or a Lewis acidic cation.

18. The method of claim 7 wherein M is Lu(III), Eu(III), Dy(III), or Tb(III).

19. The method of claim 8 where the site-directed molecule is an oligonucleotide.

20. The method of claim 8 where the site-directed molecule is an oligodeoxyribonucleotide.

21. The method of claim 19 where the oligonucleotide has complementary binding affinity for oncogenes.

22. The method of claim 19 where the phosphate ester is RNA and the oligonucleotide has complementary binding affinity for the RNA in a region proximal to the phosphate ester bond being hydrolyzed.

23. The method of claim 22 where the RNA is viral RNA.

24. The method of claim 22 where the RNA is retroviral RNA.

25. The method of claim 19 where the oligonucleotide has complementary binding affinity for a bacterial nucleic acid.

26. The method of claim 25 where the bacterial nucleic acid is ribosomal RNA.

27. The method of claim 8 where R₅ or R₆ is an oligonucleotide or a couple to an oligonucleotide.

28. The method of claim 8 where R₃ is an oligonucleotide or is a couple to an oligonucleotide.

29. The method of claim 7 where R₁ is (CH₂)₂ CH₂ OH, R₂ and R₃ are CH₂ CH₃, R₄ is CH₃, and R₅ and R₆ are OCH₂ CH₂ CH₂ OH.

30. The method of claim 7 where R₁ is (CH₂)₂ CH₂ OH, R₂ and R₃ are CH₂ CH₃, R₄ is CH₂, R₅ is O(CH₂)_n CO-oligonucleotide where n is 1-7, and R₆ is H.

31. The method of claim 30 where n is 1.

32. The method of claim 30 where n is 3.

33. The method of claim 7 where R₁ is CH₂ CH₂ CH₂ OH, R₂ and R₃ are CH₂ CH₃, R₄ is CH₃, R₅ is O(CH₂ CH₂ O)₂ CH₂ CH₂ OCH₃ and R₆ is a site-directed molecule or a couple to a site-directed molecule.

34. The method of claim 33 where R₆ is O(CH₂)_n CO-oligonucleotide where n is 1-7.

35. The method of claim 19 where the oligonucleotide has complementary binding affinity for a targeted intracellular mRNA.

36. The method of claim 35 where the mRNA is transcribed from an oncogene.

37. The method of claim 35 where the mRNA encodes a growth factor.

38. The method of claim 7 where R₁ -R₆ are as in FIG. 18 for texaphyrins A1-A24.

39. The method of claim 38 wherein M is a lanthanide cation or a Lewis acidic cation.

40. The method of claim 38 wherein M is Lu(III), Eu(III), Dy(III), or Tb(III).