Fluoresence detection using texaphyrins

• US 5,632,970 A
• Filed: 06/07/1995
• Issued: 05/27/1997
• Est. Priority Date: 01/21/1992
• Status: Expired due to Term

• Current Assignee: Pharmacyclics LLC (Abbvie Incorporated)
• Original Assignee: Pharmacyclics Incorporated (Abbvie Incorporated), Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Kr al, Vladimir A., Sessler, Jonathan L., Magda, Darren, Mody, Tarak D., Hemmi, Gregory W.
• Primary Examiner(s): Raymond, Richard L.
• Application Number:
  US08/486,967
• Time in Patent Office: 720 Days
• Field of Search: 514/185, 514/410, 424/9.6, 424/9.61
• US Class Current: 424/9.61
• 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 41/0057 Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
• 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 49/0054 Macromolecular compounds, i.e. oligomers, polymers, dendrimers
• CPC Class Codes: A61K 49/085 conjugated systems
• CPC Class Codes: A61K 49/10 Organic compounds
• 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: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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 2531/36 : Yttrium
• CPC Class Codes: B01J 2531/37 : Lanthanum
• 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 31/1805 : the ligands containing nitrogen
• 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

Texaphyrins are provided for use as radiation sensitizers. Advantageous properties of texaphyrins for use as a radiation sensitizer include: i) a low redox potential which allows radiation-induced hydrated electrons to flow to texaphyrin rather than neutralizing hydroxyl radicals, allowing hydroxyl radicals to cause cellular damage, ii) a relatively stable texaphyrin radical that reacts readily to covalently modify neighboring molecules causing further cellular damage, iii) intrinsic biolocatization, and iv) indifference to the presence or absence of O₂. These properties allow texaphyrins to be particularly effective for treating the hypoxic areas of solid neoplasms. Methods of treatment for an individual having a neoplasm or atheroma include the use of a texaphyrin as a radiation sensitizer and as an agent for photodynamic tumor therapy, or the use of a texaphyrin for internal and for external ionizing radiation. Novel texaphyrins are provided.

Claims

1. A method of determining the localization site of a water-soluble detectable texaphyrin, the method comprising the step of observing fluorescence of the texaphyrin wherein fluorescence indicates the presence of the texaphyrin.

2. The method of claim 1 where the localization site is a neoplasm or an atheroma in a host.

3. The method of claim 2 where the neoplasm is a tumor.

4. The method of claim 1 where the step of observing fluorescence is at a wavelength about 20-35 nm longer than an absorbing wavelength.

5. The method of claim 1 where the localization site is in blood, human serum, liver, atheroma, or tumor tissue.

6. The method of claim 1 wherein the texaphyrin is a water-soluble texaphyrin having at least one hydroxy substituent.

7. The method of claim 1 where the texaphyrin has structure A or B:

##STR5## R₁₁ is alkyl, alkenyl, oxyalkyl or hydroxyalkyl having up to about 3 carbon atoms and having rotational flexibility around a first-bound carbon atom;

where R₆ and R₉ are other than hydrogen, then R₅ and R₁₀ are hydrogen or methyl;

where R₅ and R₁₀ are other than hydrogen, then R₆ and R₉ are hydrogen, hydroxyl, or halide other than iodide; and

N is 0, 1 or 2.

8. The method of claim 7 where the site-directing molecule is an oligonucleotide, a peptide, protein, steroid, hormone, hormone mimic or macrocycle.

9. The method of claim 7 where R₁ is (CH₂)₂ CH₂ OH;

R₂ and R₃ are CH₂ CH₃ ;

R₄ is CH₃ ;

R₅, R₆, R₉, and R₁₀ are H;

R₇ is H, CH₃, OCH₃ or O(CH₂ CH₂ O)₂ CH₂ CH₂ OCH₃ ;

R₈ is a site-directing molecule, or a couple to a site-directing molecule; and

R₁₁ is methyl.

10. The method of claim 7 where R₄ is methyl and R₁ -R₃ and R₅ -R₁₀ are as in Table 1 for texaphyrins A1-A38.

11. The method of claim 7 where R₂ and R₃ are ethyl, R₄ is methyl, and R₁ and R₅ -R₁₁ are as in Table 2 for texaphyrins B1-B28.

12. The method of claim 7 where R₁ is (CH₂)₂ CH₂ OH;

R₂ and R₃ are CH₂ CH₃ ;

R₄ is CH₃ ;

R₅, R₆, R₉, and R₁₀ are H;

R₇ and R₈ are OCH₂ CH₂ CH₂ OH; and

R₁₁ is methyl.

13. The method of claim 7 where R₁ is CH₂ CH₂ CH₂ OH;

R₂ and R₃ are CH₂ CH₃ ;

R₄ is CH₃ ;

R₅, R₆, R₉, and R₁₀ are H;

R₇ and R₈ are O(CH₂ CH₂ O)₂ CH₂ CH₂ OCH₃ ; and

R₁₁ is methyl.

14. 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).

15. 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 or equal to ((2n+1)-q);

q is zero or a positive integer less than or equal to (2n+1); and

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 1to 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), and

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 H, alkyl, hydroxyalkyl, or saccharide.

16. 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 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), and

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 H, alkyl, hydroxyalkyl, or saccharide.

17. 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 or equal to ((2n+1)-q);

q is zero or a positive integer less than or equal to (2n+1); and

R^c is (CH₂)_n CO₂ R^d, (CH₂)_n CONHR^d, or (CH₂)_n CON(R^d)₂ where

n is a positive integer from 1 to 10, and

R^d is 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), and

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 H, alkyl, hydroxyalkyl, or saccharide.

18. The method of claim 7 where the texaphyrin has structure A, and M is a diamagnetic metal cation selected from the group consisting of Lu(III), La(III) and In(III).

19. The method of claim 7 where the texaphyrin has structure A, and M is Y(III).

20. The method of claim 7 where M is selected from the group consisting of Lu(III), La(III), In(III), and Y(III);

R₄ is methyl; and

R₁ -R₃ and R₅ -R₁₀ are as in Table 1 for texaphyrins A1-A38.

21. The method of claim 7 where the localization site is in blood, human serum, liver, atheroma, or tumor tissue.

22. A method of determining the localization site of a water-soluble detectable texaphyrin, the method comprising the step of observing fluorescence of the texaphyrin wherein fluorescence indicates the presence of the texaphyrin, the detectable texaphyrin having structure A:

##STR6## wherein, M is H, a divalent metal cation or a trivalent metal cation;

R₁ -R₄ and R₆ -R₉ are independently hydrogen, halide other than iodide, hydroxyl, alkyl, aryl, haloalkyl other than iodoalkyl, nitro, formyl, acyl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, saccharide, aminoalkyl, oxyaminoalkyl, carboxy, carboxyalkyl, carboxyamidealkyl, a site-directing molecule or a couple to a site-directing molecule;

R₅ and R₁₀ are independently hydrogen, alkyl, aryl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, aminoalkyl, oxyaminoalkyl, carboxyalkyl, carboxyamidealkyl or a couple to a site-directing molecule;

where R₆ and R₉ are other than hydrogen, then R₅ and R₁₀ are hydrogen or methyl;

where R₅ and R₁₀ are other than hydrogen, then R₆ and R₉ are hydrogen, hydroxyl, or halide other than iodide; and

N is 0, 1 or 2.

24. The method of claim 22 where R₄ is methyl and R₁ -R₃ and R₅ -R₁₀ are as in Table 1 for texaphyrins A1-A38.

25. The method of claim 22 where M is a diamagnetic metal cation.

26. The method of claim 24 where M is selected from the group consisting of Gd(III), Lu(III), La(III), In(III), and Y(III).

27. The method of claim 22 where the localization site is in blood, human serum, liver, atheroma, or tumor tissue.

28. The method of claim 22 where R₁ is (CH₂)₂ CH₂ OH;

each of R₂ and R₃ is CH₂ CH₃ ;

R₄ is CH₃ ;

each of R₅, R₆, R₉, and R₁₀ is H; and

each of R₇ and R₈ is O(CH₂)₂ CH₂ OH, or each of R₇ and R₈ is O(CH₂ CH₂)₂ CH₂ CH₂ OCH₃, or R₇ is H, CH₃, OCH₃ or O(CH₂ CH₂ O)₂ CH₂ CH₂ OCH₃ and R₈ is a site-directing molecule or a couple to a site-directing molecule.

29. The method of claim 22 where the site-directing molecule is an oligonucleotide, a peptide, a protein, asteroid, a hormone, a hormone mimic or a macrocycle.

30. The method of claim 22 where M is Lu(III) or Y(III);

R₁ is (CH₂)₂ CH₂ OH;

each of R₂ and R₃ is CH₂ CH₃ ;

R₄ is CH₃ ;

each of R₅, R₆, R₉, and R₁₀ is H; and

each of R₇ and R₈ is O(CH₂ CH₂ O)₂ CH₂ CH₂ OCH₃.

31. The method of claim 22 where M is Lu(III) or Y(III);

R₁ is (CH₂)₂ CH₂ OH;

each of R₂ and R₃ is CH₂ CH₃ ;

R₄ is CH₃ ;

each of R₅, R₆, R₉, and R₁₀ is H;

R₇ is H, CH₃, or OCH₃ ; and

R₈ is an oligonucleotide or a couple to an oligonucleotide.

36. The method of claim 22 where M is selected from the group consisting of Gd(III), Lu(III), La(III), In(III), and Y(III).

37. The method of claim 28 where M is selected from the group consisting of Gd(III), Lu(III), La(III), In(III), and Y(III).

23. A method of determining the localization site of a water-soluble detectable texaphyrin, the method comprising the step of observing fluorescence of the texaphyrin wherein fluorescence indicates the presence of the texaphyrin, the detectable texaphyrin having structure B:

##STR7## wherein, R₁ -R₄ and R₆ -R₉ are independently hydrogen, halide other than iodide, hydroxyl alkyl, aryl, haloalkyl other than iodoalkyl, nitro, formyl, acyl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, saccharide, aminoalkyl, oxyaminoalkyl, carboxy, carboxyalkyl, carboxyamidealkyl, a site-directing molecule or a couple to a site-directing molecule;

R₅ and R₁₀ are independently hydrogen, alkyl, aryl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, aminoalkyl, oxyaminoalkyl, carboxyalkyl, carboxyamidealkyl or a couple to a site-directing molecule;

R₁₁ is alkyl, alkenyl, oxyalkyl or hydroxyalkyl having up to about 3 carbon atoms and having rotational flexibility around a first-bound carbon atom;

where R₆ and R₉ are other than hydrogen, then R₅ and R₁₀ are hydrogen or methyl; and

where R₅ and R₁₀ are other than hydrogen, then R₆ and R₉ are hydrogen, hydroxyl, or halide other than iodide.

32. The method of claim 23 where R₂ and R₃ are ethyl, R₄ is methyl, and R₁ and R₅ -R₁₁ are as in Table 2 for texaphyrins B1-B28.

33. The method of claim 23 where the localization site is in blood, human serum, liver, atheroma, or tumor tissue.

34. The method of claim 23 where R₁ is (CH₂)₂ CH₂ OH;

each of R₂ and R₃ is CH₂ CH₃ ;

R₄ is CH₃ ;

each of R₅, R₆, R₉, and R₁₀ is H;

each of R₇ and R₈ is O(CH₂)₂ CH₂ OH, or each of R₇ and R₈ is O(CH₂ CH₂ O)₂ CH₂ CH₂ OCH₃, or R₇ is H, CH₃, OCH₃ or O(CH₂ CH₂ O)₂ CH₂ CH₂ OCH₃ and R₈ is a site-directing molecule or a couple to a site-directing molecule; and

R₁₁ is CH₃.

35. The method of claim 23 where the site-directing molecule is an oligonucleotide, a peptide, a protein, asteroid, a hormone, a hormone mimic or a macrocycle.

38. A method of determining the localization site of a detectable texaphyrin, the method comprising the step of observing fluorescence of the texaphyrin wherein fluorescence indicates the presence of the texaphyrin, the detectable texaphyrin having the structure:

##STR8## wherein, M is selected from the group consisting of Gd(III), Lu(III), La(III), In(III), and Y(III).

39. The method of claim 38 where the localization site is in blood, human serum, liver, atheroma, or tumor tissue.

40. The method of claim 38 where M is Gd(III).

41. The method of claim 38 where M is Lu(III).

42. The method of claim 38 where M is Y(III).