Methods of inhibiting and treating bacterial biofilms by metal chelators

• US 20050143286A1
• Filed: 10/04/2004
• Published: 06/30/2005
• Est. Priority Date: 04/18/2002
• Status: Active Grant

• Current Assignee: University of Iowa Research Foundation
• Original Assignee: University of Iowa Research Foundation
• Inventors: Greenberg, Everett P., Singh, Pradeep K., Welsh, Michael J.
• Granted Patent: US 7,446,089 B2
• Time in Patent Office: Days
• US Class Current: 514/2.800
• CPC Class Codes: A01N 61/00 Biocides, pest repellants or attractants, or plant growth regulators containing substances of unknown or undetermined composition, e.g. substances characterised only by the mode of action
• CPC Class Codes: A01N 63/50 Isolated enzymes; Isolated proteins peptides A01N37/46
• CPC Class Codes: A23B 4/20 Organic compounds; Microorganisms; Enzymes
• CPC Class Codes: A23B 4/22 Microorganisms; Enzymes; Antibiotics
• CPC Class Codes: A23L 3/34635 Antibiotics
• CPC Class Codes: A23L 3/3526 Organic compounds containing nitrogen
• CPC Class Codes: A23L 3/3571 Microorganisms; Enzymes
• CPC Class Codes: A61K 38/40 Transferrins, e.g. lactoferrins, ovotransferrins
• CPC Class Codes: A61L 2/0082 using chemical substances
• CPC Class Codes: A61L 2/16 using chemical substances
• CPC Class Codes: A61L 2/186 Peroxide solutions
• CPC Class Codes: A61L 2202/24 Medical instruments, e.g. endoscopes, catheters, sharps
• CPC Class Codes: A61P 31/04 Antibacterial agents

Abstract

The invention presented herein provides methods and compositions for the prevention and treatment of bacterial infections. The methods are based on the discovery that depletion of bioavailable iron stimulates surface motility in bacteria thus inhibiting the ability of a bacterial population to develop into a biofilm.

Claims

1. A method of inhibiting biofilm formation by bacteria comprising contacting said bacteria with an effective amount of a metal chelator, to thereby inhibit biofilm formation by said bacteria.

2. The method of claim 1, wherein said metal chelator is a proteinaceous metal chelator.

3. The method of claim 1, wherein said metal chelator is a non-proteinaceous metal chelator.

4. The method of claim 1, wherein the proteinaceous metal chelator is an iron chelator.

5. The method of claim 4, wherein the iron chelator is a naturally occurring polypeptide or fragment thereof.

6. The method of claim 4, wherein the iron chelator is an engineered polypeptide.

7. The method of claim 4, wherein the iron chelator is lactoferrin.

8. The method of claim 4, wherein the iron chelator is conalbumin.

9. The method of claim 1, wherein the bacteria are from the genus Pseudomonas.

10. The method of claim 9, wherein the pseudomonas is from the species aeruginosa.

74. The method of anyone of claims 1, 11, 22, 33, 47, or 68, wherein the bacteria are contacted with said metal chelator prior to a point at which the bacteria stop roaming as individuals.

75. The method of claim 74, wherein said metal chelator is a proteinaceous metal chelator.

76. The method of claim 74, wherein said metal chelator is a non-proteinaceous metal chelator.

100. The method of claim 3, wherein the non-proteinaceous metal chelator is a small molecule metal chelator excluding EDTA, EGTA, deferoxamine, detheylenetriaminepenta acetic acid and etidronate.

101. The method of claim 3, wherein the non-proteinaceous metal chelator is ethylenediamine-N,N,N′

,N′

-tetraacetic acid (EDTA);

the disodium, trisodium, tetrasodium, dipotassium, tripotassiun, dilithium and diammonium salts of EDTA;

the barium, calcium, cobalt, copper, dysprosium, europium, iron, indium, lanthanum, magnesium, manganese, nickel, samarium, strontium, and zinc chelates of EDTA;

trans-1,2-diaminocyclohexane-N,N,N′

,N′

-tetraaceticacid monohydrate;

N,N-bis(2-hydroxyethyl)glycine;

1,3-diamino-2-hydroxypropane-N,N,N′

,N′

-tetraacetic acid;

1,3-diaminopropane-N,N,N′

,N′

-tetraacetic acid;

ethylenediamine-N,N′

-diacetic acid;

ethylenediamine-N,N′

-dipropionic acid dihydrochloride;

ethylenediamine-N,N′

-bis(methylenephosphonic acid)hemihydrate;

N-(2-hydroxyethyl)ethylenediamine-N,N′

,N′

-triacetic acid;

ethylenediamine-N,N,N′

,N′

-tetrakis(methylenephosponic acid);

O,O′

-bis(2-aminoethyl)ethyleneglycol-N,N,N′

,N′

-tetraacetic acid;

N,N-bis(2-hydroxybenzyl) ethylene diamine-N,N-diacetic acid;

1,6-hexamethylenediamine-N,N,N′

,N′

-tetraacetic acid;

N-(2-hydroxyethyl)iminodiacetic acid;

iminodiacetic acid;

1,2-diaminopropane-N,N,N′

,N′

-tetraacetic acid;

nitrilotriacetic acid;

nitrilotripropionic acid;

the trisodium salt of nitrilotris(methylenephosphoric acid);

7,19,30-trioxa-1,4,10,13,16,22,27,33-octaazabicyclo[11,11,11]pentatriacontane hexahydrobromide;

or;

triethylenetetrainie-N,N,N′

,N″

,N′

″

,N′

″

-hexaacetic acid.

11. A method of stimulating surface motility in bacteria comprising contacting said bacteria with a metal chelator.

12. The method of claim 11 wherein said metal chelator is a proteinaceous metal chelator.

13. The method of claim 11 wherein said metal chelator is a non-proteinaceous metal chelator.

14. The method of claim 11, wherein bioavailable iron is limited by said proteinaceous metal chelator, thereby stimulating surface motility in said bacteria.

15. The method of claim 12, wherein the proteinaceous metal chelator is an iron chelator.

16. The method of claim 15, wherein the iron chelator is a naturally occurring polypeptide or fragment thereof.

17. The method of claim 15, wherein the iron chelator is an engineered polypeptide.

18. The method of claim 15, wherein the iron chelator is lactofemin.

19. The method of claim 15, wherein the iron chelator is conalbumin.

20. The method of claim 11, wherein the bacteria are from the genus Pseudomonas.

21. The method of claim 20, wherein the pseudomonas is from the species aeruginosa.

61. The method of any one of claims 7, 18, 29, or 41, wherein the concentration of lactoferrin is between 5 and 50 μ

g/ml.

62. The method of any one of claims 7, 18, 29, or 41, wherein the concentration of lactoferrin is between 15 and 40 μ

g/ml.

63. The method of any one of claims 7, 18, 29, or 41, wherein the concentration of lactoferrin is between 20 and 30 μ

g/ml.

64. The method of any one of claims 7, 18, 29, or 41, wherein the concentration of lactoferrin is 20 μ

g/ml.

22. A method of inhibiting biofilm development by bacteria in a subject comprising administering to said subject an effective amount of a metal chelator, thereby inhibiting biofilm development by said bacteria in said subject.

23. The method of claim 22, wherein said metal chelator is a proteinaceous metal chelator.

24. The method of claim 22, wherein said metal chelator is a non-proteinaceous metal chelator.

25. The method of claim 22, wherein the biofilm development is in a region selected from the group of the airway, cornea, middle ear, heart, prostate, kidney, and bone.

26. The method of claim 22, wherein the proteinaceous metal chelator is an iron chelator.

27. The method of claim 26, wherein the iron chelator is a naturally occurring polypeptide or fragment thereof.

28. The method of claim 26, wherein the iron chelator is an engineered polypeptide.

29. The method of claim 26, wherein the iron chelator is lactoferrin.

30. The method of claim 26, wherein the iron chelator is conalbumin.

31. The method of claim 26, wherein the bacteria are from the genus Pseudomonas.

32. The method of claim 31, wherein the pseudomonas is from the species aeruginosa.

33. A method of treating a subject suffering from a bacterial infection comprising administering to said subject a composition comprising a therapeutically effective amount of a metal chelator, thereby treating the subject suffering from a bacterial infection.

34. The method of claim 33, wherein said metal chelator is a proteinaceous metal chelator.

35. The method of claim 33, wherein said metal chelator is a non-proteinaceous metal chelator.

36. The method of claim 33, wherein said composition further comprises an effective amount of an antibacterial agent.

37. The method of claim 33, wherein said antibacterial agent is an antibiotic.

38. The method of claim 33, wherein the proteinaceous metal chelator is an iron chelator.

39. The method of claim 38, wherein the proteinaceous iron chelator is a naturally occurring polypeptide or fragment thereof.

40. The method of claim 38, wherein the iron chelator is an engineered polypeptide.

41. The method of claim 38, wherein the iron chelator is lactoferrin.

42. The method of claim 38, wherein the iron chelator is conalbumin.

43. The method of claim 33, wherein the bacteria are from the genus Pseudomonas.

44. The method of claim 43, wherein the pseudomonas is from the species aeruginosa.

45. The method of claim 37, wherein the antibiotic is tobramycin, tazobactam, ciprofloxin, piperacillin, semi-synthetic penicillins, amino glycosides, fluoroquinones, cephlosporins, or clindamycins.

46. The composition of claim 43, wherein the antibiotic is selected from the group consisting of tobramycin, tazobactam, ciprofloxin, piperacillin and clindamycin.

47. A method of inhibiting biofilm development on a medical indwelling device in a subject comprising administering to said subject an effective amount of a metal chelator, thereby inhibiting biofilm development on said medical indwelling device.

48. The method of claim 47, wherein said metal chelator is a proteinaceous metal chelator.

49. The method of claim 47, wherein said metal chelator is a non-proteinaceous metal chelator.

50. The method of claim 47, wherein said device is selected from the group consisting of catheters, orthopedic devices, prosthetic devices, vascular stents, urinary stents, pacemakers, and implants.

51. A microbial disinfectant composition comprising an effective amount of a metal chelator.

52. The microbial disinfectant of claim 51, wherein in the metal chelator is a proteinaceous metal chelator.

53. The microbial disinfectant of claim 51, wherein in the metal chelator is a non-proteinaceous metal chelator.

54. The disinfectant of claim 51, further comprising a biocide.

55. The disinfectant of claim 51, further comprising an antibiotic.

56. A pharmaceutical composition comprising a therapeutically effective amount of a metal chelator and a pharmaceutically acceptable carrier.

57. The pharmaceutical composition of claim 56, wherein in the metal chelator is a proteinaceous metal chelator.

58. The pharmaceutical composition of claim 56, wherein in the metal chelator is a non-proteinaceous metal chelator.

59. The pharmaceutical composition of claim 56, further comprising an antibacterial agent.

60. The pharmaceutical composition of claim 59, wherein said antibacterial agent is an antibiotic.

65. A method of inhibiting biofilm formation on a piece of equipment by bacteria comprising contacting said bacteria with an effective amount of a metal chelator, to thereby inhibit biofilm formation by said bacteria.

66. The method of claim 65, wherein said metal chelator is a proteinaceous metal chelator.

67. The method of claim 65, wherein said metal chelator is a non-proteinaceous metal chelator.

68. The method of 65, wherein the equipment is used for a medical, industrial or commercial application.

69. A method of inhibiting biofilm formation on a device comprising,

contacting said device with a compound comprising an effective amount of a metal chelator,

thereby inhibiting the formation of biofilm on said device.

70. The method of claim 69, wherein said metal chelator is a proteinaceous metal chelator.

71. The method of claim 69, wherein said metal chelator is a non-proteinaceous metal chelator.

72. The method of claim 69, wherein contacting comprises bathing said device in a solution comprising an effective amount of a proteinaceous metal chelator.

73. The method of claim 69, wherein contacting comprises coating said device in a solution comprising an effective amount of a proteinaceous metal chelator.

77. A kit for treating a bacterial infection comprising a metal chelator and directions for use.

78. The kit of claim 77, wherein said metal chelator is a proteinaceous metal chelator.

79. The kit of claim 77, wherein said metal chelator is a non-proteinaceous metal chelator.

80. The kit of claim 77, further comprising an antimicrobial agent.

81. The kit of claim 80, wherein the antimicrobial agent is an antibiotic.

82. A method of inhibiting biofilm formation by bacteria comprising

obtaining a metal chelator;

and

contacting said bacteria with an effective amount of a metal chelator

to thereby inhibit biofilm formation by said bacteria.

83. The method of claim 82, wherein said metal chelator is a proteinaceous metal chelator.

84. The method of claim 82, wherein said metal chelator is a non-proteinaceous metal chelator.

85. A method of stimulating surface motility in bacteria comprising

obtaining a metal chelator and

contacting said bacteria with the metal chelator

thereby stimulating surface motility in bacteria.

86. The method of claim 85, wherein said metal chelator is a proteinaceous metal chelator.

87. The method of claim 85 wherein said metal chelator is a non-proteinaceous metal chelator.

88. A method of inhibiting biofilm development by bacteria in a subject comprising obtaining a metal chelator;

and

administering to said subject an effective amount of said metal chelator thereby inhibiting biofilm development by said bacteria in said subject.

89. The method of claim 88, wherein said metal chelator is a proteinaceous metal chelator.

90. The method of claim 88, wherein said metal chelator is a non-proteinaceous metal chelator.

91. A method of treating a subject suffering from a bacterial infection comprising

obtaining a metal chelator;

and

administering to said subject a composition comprising an effective amount of said metal chelator

thereby treating the subject suffering from a bacterial infection.

92. The method of claim 91, wherein said metal chelator is a proteinaceous metal chelator.

93. The method of claim 91 wherein said metal chelator is a non-proteinaceous metal chelator.

94. A method of inhibiting biofilm development on a medical indwelling device in a subject comprising

obtaining a metal chelator;

and

administering to said subject an effective amount of said metal chelator thereby inhibiting biofilm development on said medical indwelling device.

95. The method of claim 94, wherein said metal chelator is a proteinaceous metal chelator.

96. The method of claim 94, wherein said metal chelator is a non-proteinaceous metal chelator.

97. A method of inhibiting biofihm formation on food comprising,

contacting said food with a compound comprising an effective amount of a metal chelator

thereby inhibiting the formation of biofilm on said food.

98. The method of claim 97, wherein said food is further contacted with an antibiotic.

99. The method of claim 98, wherein said food is meat.