Claims
- 1. A method for identifying a molecule that binds an irradiated tumor in a subject, the method comprising:
(a) exposing a tumor to ionizing radiation; (b) administering to a subject a library of diverse molecules; and (c) isolating one or more molecules of the library from the tumor, whereby a molecule that binds an irradiated tumor is identified.
- 2. The method of claim 1, wherein the exposing comprises exposing the tumor to less than about 2 Gy ionizing radiation.
- 3. The method of claim 1, wherein the exposing comprises exposing the tumor to at least about 2 Gy ionizing radiation.
- 4. The method of claim 3, wherein the exposing comprises exposing the tumor to about 10 Gy to about 20 Gy ionizing radiation.
- 5. The method of claim 1, wherein the administering further comprises administering the library to isolated tumor cells or to isolated proteins prior to administering the library to the subject.
- 6. The method of claim 1, wherein the administering comprises administering the library by intravascular provision.
- 7. The method of claim 1, wherein the administering comprises administering the library subsequent to radiation exposure.
- 8. The method of claim 7, wherein the administering comprises administering the library 0 hours to about 24 hours following radiation exposure.
- 9. The method of claim 8, wherein the administering comprises administering the library about 4 hours to about 24 hours following radiation exposure.
- 10. The method of claim 1, wherein the subject is a warm-blooded vertebrate.
- 11. The method of claim 10, wherein the subject is a human.
- 12. The method of claim 1, wherein the library of diverse molecules comprises a library of ten or more diverse molecules.
- 13. The method of claim 12, wherein the library of diverse molecules comprises a library of one hundred or more diverse molecules.
- 14. The method of claim 13, wherein the library of diverse molecules comprises a library of a billion or more diverse molecules.
- 15. The method of claim 1, wherein the library of diverse molecules comprises a library of molecules selected from the group consisting of peptides, peptide mimetics, proteins, antibodies or fragments thereof, small molecules, nucleic acids, and combinations thereof.
- 16. The method of claim 15, wherein the library of diverse molecules comprises a library of peptides, antibodies, or a combination thereof.
- 17. The method of claim 1, wherein the molecule that binds an irradiated tumor comprises a ligand that binds a tumor cell, an endothelial cell associated with tumor vasculature, or a blood component.
- 18. The method of claim 1, wherein each of the exposing, administering, and isolating is repeated one or more times.
- 19. A molecule identified by the method of claim 1.
- 20. A method for identifying a molecule that binds an irradiated tumor in a subject comprising:
(a) exposing a tumor and a control tissue to ionizing radiation; (b) administering to the tumor and to the control tissue a library of diverse molecules; (c) detecting one or more molecules of the library that bind to the tumor and that substantially lack binding to the control tissue, whereby a molecule that binds an irradiated tumor is identified.
- 21. The method of claim 20, wherein the exposing comprises exposing the tumor to less than about 2 Gy ionizing radiation.
- 22. The method of claim 21, wherein the exposing comprises exposing the tumor to at least about 2 Gy ionizing radiation.
- 23. The method of claim 22, wherein the exposing comprises exposing the tumor to about 10 Gy to about 20 Gy ionizing radiation.
- 24. The method of claim 20, wherein the administering further comprises administering the library to isolated tumor cells or to isolated proteins prior to administering the library to the subject.
- 25. The method of claim 20, wherein the library of diverse molecules comprises a library of ten or more diverse molecules.
- 26. The method of claim 25, wherein the library of diverse molecules comprises a library of one hundred or more diverse molecules.
- 27. The method of claim 26, wherein the library of diverse molecules comprises a library of a billion or more diverse molecules.
- 28. The method of claim 20, wherein the library of diverse molecules comprises a library of molecules selected from the group consisting of peptides, peptide mimetics, proteins, antibodies or fragments thereof, small molecules, nucleic acids, and combinations thereof.
- 29. The method of claim 28, wherein the library of diverse molecules comprises a library of peptides, antibodies, or a combination thereof.
- 30. The method of claim 20, wherein the molecule that binds an irradiated tumor comprises a ligand that binds a tumor cell, an endothelial cell associated with tumor vasculature, or a blood component.
- 31. The method of claim 20, further comprising:
(a) isolating the tumor and the control tissue, or fractions thereof; and (b) administering the library to the isolated tumor and to the control tissue, or fractions thereof, in vitro.
- 32. A molecule identified by the method of claim 20.
- 33. A method for preparing a composition for x-ray-guided drug delivery, the method comprising:
(a) performing in vivo panning of claim 1, whereby a targeting ligand that binds a radiation-inducible tumor molecule is identified; and (b) conjugating the targeting ligand to a drug, whereby a composition for x-ray-guided drug delivery is prepared.
- 34. A method for preparing a composition for x-ray-guided drug delivery, the method comprising:
(a) performing in vivo panning of claim 20, whereby a targeting ligand that binds a radiation-inducible tumor molecule is identified; and (b) conjugating the targeting ligand to a drug, whereby a composition for x-ray-guided drug delivery is prepared.
- 35. A composition for x-ray guided drug delivery comprising one or more targeting ligands identified by in vivo panning, wherein the one or more targeting ligands each bind to an irradiated tumor.
- 36. The composition of claim 35, wherein the one or more targeting ligands comprises a peptide comprising an amino acid sequence of any one of SEQ ID NOs:1-5, or combinations thereof.
- 37. The composition of claim 35, wherein the one or more targeting ligands comprises a peptide comprising an amino acid sequence of any one of SEQ ID NOs:1-13, or combinations thereof.
- 38. The composition of claim 35, wherein the one or more targeting ligands comprises a single chain antibody comprising:
(a) a polypeptide comprising an amino acid sequence of SEQ ID NO:18 or 20; (b) a polypeptide comprising an amino acid sequence substantially identical to SEQ ID NO:18 or 20; (c) a polypeptide encoded by a nucleotide sequence of SEQ ID NO:17 or 19; (d) a polypeptide encoded by a nucleotide sequence substantially identical to SEQ ID NO:17 or 19; or (e) combinations thereof.
- 39. The composition of claim 35, wherein the irradiated tumor comprises a tumor exposed to less than about 2 Gy ionizing radiation.
- 40. The composition of claim 39, wherein the irradiated tumor comprises a tumor exposed to at least about 2 Gy ionizing radiation.
- 41. The composition of claim 40, wherein the irradiated tumor comprises a tumor exposed to about 10 Gy to about 20 Gy ionizing radiation.
- 42. The composition of claim 35, wherein the targeting ligand binds to an irradiated glioma, a melanoma, or lung carcinoma.
- 43. The composition of claim 35, wherein the targeting ligand binds to an irradiated tumor of two or more tumor types.
- 44. The composition of claim 43, wherein the targeting ligand binds to an irradiated tumor of three or more tumor types.
- 45. The composition of claim 44, wherein the targeting ligand binds to an irradiated glioma, a melanoma, and a lung carcinoma.
- 46. The composition of claim 35, further comprising a detectable label.
- 47. The composition of claim 46, wherein the detectable label is detectable in vivo.
- 48. The composition of claim 47, wherein the detectable label comprises a label that can be detected using magnetic resonance imaging, scintigraphic imaging, ultrasound, or fluorescence.
- 49. The composition of claim 48, wherein the label that can be detected using scintigraphic imaging comprises a radionuclide label.
- 50. The composition of claim 49, wherein the radionuclide label comprises 131I or 99mTc.
- 51. The composition of claim 35, further comprising a therapeutic agent.
- 52. The composition of claim 51, wherein the therapeutic agent is selected from the group consisting of a radionuclide, a cytotoxin, a therapeutic gene, and a chemotherapeutic agent.
- 53. The composition of claim 35, further comprising a drug carrier.
- 54. The composition of claim 53, wherein the drug carrier is selected from the group consisting of a viral vector, a liposome, a plasmid, a microcapsule, and combinations thereof.
- 55. The composition of claim 54, wherein the drug carrier comprises a liposome or a microsphere.
- 56. The composition of claim 51, further comprising a detectable label.
- 57. The composition of claim 56, wherein the detectable label comprises a label that can be detected using magnetic resonance imaging, scintigraphic imaging, ultrasound, or fluorescence.
- 58. The composition of claim 57, wherein the label that can be detected using scintigraphic imaging comprises a radionuclide label.
- 59. The composition of claim 58, wherein the radionuclide label comprises 131I or 99mTc.
- 60. A method for detecting a tumor in a subject comprising:
(a) exposing a suspected tumor to ionizing radiation; (b) administering to the subject one or more targeting ligands identified by in vivo panning, wherein the one or more targeting ligands binds to an irradiated tumor, and wherein the one or more targeting ligands comprises a detectable label; and (c) detecting the detectable label, whereby a tumor is diagnosed.
- 61. The method of claim 60, wherein the exposing comprises exposing the tumor to less than about 2 Gy ionizing radiation.
- 62. The method of claim 61, wherein the exposing comprises exposing the tumor to at least about 2 Gy ionizing radiation.
- 63. The method of claim 62, wherein the exposing comprises exposing the tumor to about 10 Gy to about 20 Gy ionizing radiation.
- 64. The method of claim 61, wherein the administering comprises administering the targeting ligand by intravascular provision.
- 65. The method of claim 60, wherein the administering comprises administering the targeting ligand subsequent to radiation exposure.
- 66. The method of claim 65, wherein the administering comprises administering the targeting ligand 0 hours to about 24 hours following radiation exposure.
- 67. The method of claim 66, wherein the administering comprises administering the targeting ligand about 4 hours to about 24 hours following radiation exposure.
- 68. The method of claim 60, wherein the subject is a warm-blooded vertebrate.
- 69. The method of claim 68, wherein the subject is a human.
- 70. The method of claim 60, wherein the one or more targeting ligands comprises a peptide comprising an amino acid sequence of any one of SEQ ID NOs:1-5, or combinations thereof.
- 71. The method of claim 60, wherein the one or more targeting ligands comprises a peptide comprising an amino acid sequence of any one of SEQ ID NOs:1-13, or combinations thereof.
- 72. The method of claim 60, wherein the one or more targeting ligands comprises a single chain antibody comprising:
(a) a polypeptide comprising an amino acid sequence of SEQ ID NO:18 or 20; (b) a polypeptide comprising an amino acid sequence substantially identical to SEQ ID NO:18 or 20; (c) a polypeptide encoded by a nucleotide sequence of SEQ ID NO:17 or 19; (d) a polypeptide encoded by a nucleotide sequence substantially identical to SEQ ID NO:17 or 19; or (e) combinations thereof.
- 73. The method of claim 60, wherein the detectable label is detectable in vivo.
- 74. The method of claim 73, wherein the detectable label comprises a label that can be detected using magnetic resonance imaging, scintigraphic imaging, ultrasound, or fluorescence.
- 75. The method of claim 74, wherein the label that can be detected using scintigraphic imaging comprises a radionuclide label.
- 76. The method of claim 75, wherein the radionuclide label is 131I or 99mTc.
- 77. The method of claim 75, wherein the detecting comprises detecting the radionuclide label using positron emission tomography, single photon emission computed tomography, gamma camera imaging, or rectilinear scanning.
- 78. The method of claim 60, wherein the tumor is a primary or a metastasized tumor.
- 79. The method of claim 60, wherein the tumor comprises a tumor selected from the group consisting of bladder carcinoma, breast carcinoma, cervical carcinoma, cholangiocarcinoma, colorectal carcinoma, gastric sarcoma, glioma, lung carcinoma, lymphoma, melanoma, multiple myeloma, osteosarcoma, ovarian carcinoma, pancreatic carcinoma, prostate carcinoma, stomach carcinoma, a head, a neck tumor, and a solid tumor.
- 80. The method of claim 79, wherein the tumor is selected from the group consisting of a glioma, a melanoma, and a lung carcinoma.
- 81. The method of claim 60, further comprising simultaneously detecting two or more tumors in a subject.
- 82. The method of claim 81, wherein the two or more tumors in a subject comprise two or more tumor types.
- 83. A method for diagnosing a tumor in a subject comprising:
(a) exposing a suspected tumor to ionizing radiation; (b) isolating the suspected tumor, or fraction thereof; (c) contacting the suspected tumor with one or more targeting ligands identified by in vivo panning, wherein the one or more targeting ligands binds to an irradiated tumor, and wherein the one or more targeting ligands comprises a detectable label; and (d) detecting the detectable label, whereby the tumor is diagnosed.
- 84. The method of claim 83, wherein the exposing comprises exposing the tumor to less than about 2 Gy ionizing radiation.
- 85. The method of claim 84, wherein the exposing comprises exposing the tumor to about at least about 2 Gy ionizing radiation.
- 86. The method of claim 85, wherein the exposing comprises exposing the tumor to about 10 Gy to about 20 Gy ionizing radiation.
- 87. The method of claim 83, wherein the subject is a warm-blooded vertebrate.
- 88. The method of claim 87, wherein the subject is a human.
- 89. The method of claim 83, wherein the isolating comprises biopsing a tissue suspected of comprising a tumor.
- 90. The method of claim 83, wherein the isolating is performed prior to or subsequent to exposing the tissue to ionizing radiation.
- 91. The method of claim 83, wherein the one or more targeting ligands binds to an irradiated tumor.
- 92. The method of claim 83, wherein the one or more targeting ligands comprises a peptide comprising an amino acid sequence of any one of SEQ ID NOs:1-5, or combinations thereof.
- 93. The method of claim 83, wherein the one or more targeting ligands comprises a single chain antibody comprising:
(a) a polypeptide comprising an amino acid sequence of SEQ ID NO:18 or 20; (b) a polypeptide comprising an amino acid sequence substantially identical to SEQ ID NO:18 or 20; (c) a polypeptide encoded by a nucleotide sequence of SEQ ID NO:17 or 19; (d) a polypeptide encoded by a nucleotide sequence substantially identical to SEQ ID NO:17 or 19; or (e) combinations thereof.
- 94. The method of claim 83, wherein the detectable label comprises a fluorophore, an epitope, or a radiolabel.
- 95. The method of claim 83, wherein the tumor is a primary or a metastasized tumor.
- 96. The method of claim 83, wherein the tumor comprises a tumor selected from the group consisting of bladder carcinoma, breast carcinoma, cervical carcinoma, cholangiocarcinoma, colorectal carcinoma, gastric sarcoma, glioma, lung carcinoma, lymphoma, melanoma, multiple myeloma, osteosarcoma, ovarian carcinoma, pancreatic carcinoma, prostate carcinoma, stomach carcinoma, a head tumor, a neck tumor, and a solid tumor.
- 97. The method of claim 96, wherein the tumor is selected from the group consisting of a glioma, a melanoma, and a lung carcinoma.
- 98. A method for x-ray-guided delivery of a therapeutic composition, a diagnostic composition, or a combination thereof, to a tumor in a subject, the method comprising:
(a) exposing the tumor to ionizing radiation; and (b) administering to the subject a therapeutic composition, a diagnostic composition, or a combination thereof, wherein the composition comprises one or more targeting ligands identified by in vivo panning; whereby the therapeutic composition, the diagnostic composition, or the combination thereof, is selectively targeted to the tumor.
- 99. The method of claim 98, wherein the tumor is a primary or a metastasized tumor.
- 100. The method of claim 98, wherein the tumor is selected from a tumor selected from the group consisting of bladder carcinoma, breast carcinoma, cervical carcinoma, cholangiocarcinoma, colorectal carcinoma, gastric sarcoma, glioma, lung carcinoma, lymphoma, melanoma, multiple myeloma, osteosarcoma, ovarian carcinoma, pancreatic carcinoma, prostate carcinoma, stomach carcinoma, a head tumor, a neck tumor, and a solid tumor.
- 101. The method of claim 100, wherein the tumor is selected from the group consisting of a glioma, a melanoma, and a lung carcinoma.
- 102. The method of claim 98, wherein the exposing comprises exposing the tumor to less than about 2 Gy ionizing radiation.
- 103. The method of claim 102, wherein the exposing comprises exposing the tumor to at least about 2 Gy ionizing radiation.
- 104. The method of claim 103, wherein the exposing comprises exposing the tumor to about 10 Gy to about 20 Gy ionizing radiation.
- 105. The method of claim 98, wherein the administering comprises administering the targeting ligand by intravascular provision.
- 106. The method of claim 98, wherein the administering comprises administering the targeting ligand subsequent to radiation exposure.
- 107. The method of claim 106, wherein the administering comprises administering the targeting ligand 0 hours to about 24 hours following radiation exposure.
- 108. The method of claim 107, wherein the administering comprises administering the targeting ligand about 4 hours to about 24 hours following radiation exposure.
- 109. The method of claim 98, wherein the subject is a warm-blooded vertebrate.
- 110. The method of claim 109, wherein the subject is a human.
- 111. The method of claim 98, wherein the therapeutic composition further comprises a radionuclide, a cytotoxin, a therapeutic gene, or a chemotherapeutic agent.
- 112. The method of claim 98, wherein the diagnostic composition further comprises a detectable label.
- 113. The composition of claim 112, wherein the detectable label is detectable in vivo.
- 114. The method of claim 113, wherein the detectable label comprises a label that can be detected using magnetic resonance imaging, scintigraphic imaging, ultrasound, or fluorescence.
- 115. The method of claim 114, wherein the label that can be detected using scintigraphic imaging comprises a radionuclide label.
- 116. The method of claim 115, wherein the radionuclide label is 131I or 99mTc.
- 117. The method of claim 115, further comprising detecting the radionuclide label using positron emission tomography, single photon emission computed tomography, gamma camera imaging, or rectilinear scanning.
- 118. The method of claim 98, wherein the one or more targeting ligands comprises a peptide comprising an amino acid sequence of any one of SEQ ID NOs:1-5, or combinations thereof.
- 119. The method of claim 98, wherein the one or more targeting ligands comprises a single chain antibody comprising:
(a) a polypeptide comprising an amino acid sequence of SEQ ID NO:18 or 20; (b) a polypeptide comprising an amino acid sequence substantially identical to SEQ ID NO:18 or 20; (c) a polypeptide encoded by a nucleotide sequence of SEQ ID NO:17 or 19; (d) a polypeptide encoded by a nucleotide sequence substantially identical to SEQ ID NO:17 or 19; or (e) combinations thereof.
- 120. The method of claim 98, wherein the selective targeting comprises targeting to an irradiated tumor in the absence of targeting to a non-irradiated tumor, to non-irradiated normal tissue, and to irradiated normal tissue.
- 121. A method for identifying a radiation-induced target molecule comprising:
(a) providing a targeting ligand identified by in vivo panning, wherein the targeting ligand binds to an irradiated tumor; (b) exposing the targeting peptide to one or more candidate target molecules; and (c) determining selective binding of the one or more candidate target molecules to the targeting ligand, whereby a radiation-induced target molecule is identified.
- 122. The method of claim 121, wherein the targeting ligand comprises a peptide comprising an amino acid sequence of any one of SEQ ID NOs:1-5, or combinations thereof.
- 123. The method of claim 121, wherein the targeting ligands comprises a single chain antibody comprising:
(a) a polypeptide comprising an amino acid sequence of SEQ ID NO:18 or 20; (b) a polypeptide comprising an amino acid sequence substantially identical to SEQ ID NO:18 or 20; (c) a polypeptide encoded by a nucleotide sequence of SEQ ID NO:17 or 19; (d) a polypeptide encoded by a nucleotide sequence substantially identical to SEQ ID NO:17 or 19; or (e) combinations thereof.
- 124. The method of claim 121, wherein the radiation-induced target molecule comprises a molecule present on a tumor cell, an endothelial cell associated with tumor vasculature, or a blood component.
- 125. A target molecule identified by the method of claim 121.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is based on and claims priority to U.S. Provisional Patent application serial No. 60/328,123, filed Oct. 3, 2001, herein incorporated by reference in its entirety.
GRANT STATEMENT
[0002] This work was supported by grants CA58508, CA70937, CA89888, CA89674, and CA90949 from the U.S. National Institute of Health. Thus, the U.S. government has certain rights in the invention.
Provisional Applications (1)
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Number |
Date |
Country |
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60328123 |
Oct 2001 |
US |