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US-12616751-B2 - Photosensitizing antibody-fluorophore conjugates

US12616751B2US 12616751 B2US12616751 B2US 12616751B2US-12616751-B2

Abstract

Methods of killing cells are described. In particular examples, the method includes contacting a cell having a cell surface protein with a therapeutically effective amount of an antibody-IR700 molecule, wherein the antibody specifically binds to the cell surface protein, such as a tumor-specific antigen on the surface of a tumor cell. The cell is subsequently irradiated, such as at a wavelength of 660 to 740 nm at a dose of at least 1 J cm −2 . The cell is also contacted with one or more therapeutic agents (such as an anti-cancer agent), for example about 0 to 8 hours after irradiating the cell, thereby killing the cell. Also provided are methods of imaging cell killing in real time, using fluorescence lifetime imaging.

Inventors

  • Hisataka Kobayashi
  • Peter Choyke

Assignees

  • THE USA, AS REPRESENTED BY THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES

Dates

Publication Date
20260505
Application Date
20220615

Claims (20)

  1. 1 . A method of treating a tumor and detecting margins of the tumor during a surgical procedure, the method comprising: a) administering a therapeutically effective amount of an antibody-IR700 conjugate to a subject having a tumor, wherein the antibody binds to a cell surface protein on the tumor; b) irradiating the tumor at a wavelength between 660-740 nm and at a dose of at least 1 J cm −2 , thereby treating the tumor in the subject; and c) detecting fluorescence of the antibody-IR700 conjugate following the irradiating step to detect the tumor margins.
  2. 2 . The method of claim 1 , wherein the fluorescence of the antibody-IR700 conjugate is detected at an emission wavelength of 665-740 nm.
  3. 3 . The method of claim 1 , wherein the surgical procedure is an endoscopic procedure.
  4. 4 . The method of claim 1 , wherein the surgical procedure is a surgical resection of the tumor and the irradiating step is performed after the surgical resection of the tumor.
  5. 5 . The method of claim 4 , wherein the resected tumor is a tumor of the brain, breast, bone, cervix, colon, head or neck, liver, lung, ovary, pancreas, prostate, skin, stomach, or uterus.
  6. 6 . The method of claim 1 , wherein the cell surface protein is selected from the group consisting of HER1, HER2, CD20, CD25, CD33, CD52, Lewis Y, CEA, and prostate specific membrane antigen (PSMA).
  7. 7 . The method of claim 1 , wherein the antibody is cetuximab, trastuzumab, pertuzumab, rituximab, panitumumab, basiliximab, or J591, or an antigen binding fragment thereof.
  8. 8 . The method of claim 1 , wherein detecting fluorescence of the antibody-IR700 conjugate following the irradiating step further comprises irradiating the tumor at a wavelength between 660-740 nm at a dose below a threshold dose for cell killing.
  9. 9 . The method of claim 8 , wherein the dose below the threshold dose for cell killing is at least 1/1000 or at least 1/10,000 of the threshold dose for cell killing.
  10. 10 . The method of claim 1 , further comprising detecting fluorescence of the antibody-IR700 conjugate prior to step b) to detect the tumor.
  11. 11 . The method of claim 10 , wherein detecting fluorescence of the antibody-IR700 conjugate prior to step b) further comprises irradiating the tumor at a wavelength between 660-740 nm at a dose below a threshold dose for cell killing.
  12. 12 . The method of claim 11 , wherein the dose below the threshold dose for cell killing is at least 1/1000 or at least 1/10,000 of the threshold dose for cell killing.
  13. 13 . A method of treating a tumor in a subject and monitoring the treatment of the tumor, the method comprising: a) administering a therapeutically effective amount of an antibody-IR700 conjugate to the subject, wherein the antibody specifically binds to a cell surface protein on the tumor; b) irradiating the tumor at a wavelength between 660-740 nm and at a dose of at least 1 J cm −2 , thereby treating the tumor in the subject; and c) detecting fluorescence of the antibody-IR700 conjugate following the irradiating step to monitor the course of treatment.
  14. 14 . The method of claim 13 , wherein the method comprises detecting the margins of the tumor.
  15. 15 . The method of claim 13 , wherein the method comprises monitoring cell killing in real time.
  16. 16 . The method of claim 13 , wherein the antibody is cetuximab, trastuzumab, pertuzumab, rituximab, panitumumab, basiliximab, or J591, or an antigen binding fragment thereof.
  17. 17 . The method of claim 13 , wherein detecting fluorescence of the antibody-IR700 conjugate following the irradiating step further comprises irradiating the tumor at a wavelength between 660-740 nm at a dose below a threshold dose for cell killing.
  18. 18 . The method of claim 17 , wherein the dose below the threshold dose for cell killing is at least 1/1000 or at least 1/10,000 of the threshold dose for cell killing.
  19. 19 . The method of claim 13 , further comprising detecting fluorescence of the antibody-IR700 conjugate prior to step b) to detect the tumor.
  20. 20 . The method of claim 19 , wherein detecting fluorescence of the antibody-IR700 conjugate prior to step b) further comprises irradiating the tumor at a wavelength between 660-740 nm at a dose below a threshold dose for cell killing.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. application Ser. No. 16/694,761 filed Nov. 25, 2019, issued as U.S. Pat. No. 11,364,297, which is a continuation of U.S. application Ser. No. 14/868,040 filed Sep. 28, 2015, now U.S. Pat. No. 10,537,641, which is a divisional of U.S. application Ser. No. 14/126,060 filed Dec. 13, 2013, now U.S. Pat. No. 9,358,306, which is the U.S. National Stage of International Application No. PCT/US2012/044421, filed Jun. 27, 2012, which was published in English under PCT Article 21(2), which is a continuation-in-part of U.S. application Ser. No. 13/180,111 filed Jul. 11, 2011, now U.S. Pat. No. 8,524,239, which claims priority to U.S. Provisional Application No. 61/363,079, filed Jul. 9, 2010, all herein incorporated by reference. FIELD OF THE DISCLOSURE This application relates to antibody-IR700 conjugates, and methods of their use to kill cells that specifically bind to the antibody following irradiation with infrared (NIR) light. Also provided are devices that incorporate NIR light emitting diodes (LEDs) that can also be used with the disclosed conjugates and methods. BACKGROUND Cancer was responsible for about 13% of all human deaths in 2007. Although there are several therapies for cancer, there remains a need for therapies that effectively kill the tumor cells while not harming non-cancerous cells. In order to minimize the side effects of conventional cancer therapies, including surgery, radiation and chemotherapy, molecular targeted cancer therapies have been developed. Among the existing targeted therapies, monoclonal antibodies (MAb) therapy have the longest history, and to date, over 25 therapeutic MAbs have been approved by the Food and Drug Administration (FDA) (Waldmann, Nat Med 9:269-277, 2003); Reichert et al., Nat Biotechnol 23:1073-1078, 2005). Effective MAb therapy traditionally depends on three mechanisms: antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and receptor blockade and requires multiple high doses of the MAb. MAbs have also been used at lower doses as vectors to deliver therapies such as radionuclides (Goldenberg et al., J Clin Oncol 24, 823-834, 2006) or chemical or biological toxins (Pastan et al., Nat Rev Cancer 6:559-565, 2006). Ultimately, however, dose limiting toxicity relates to the biodistribution and catabolism of the antibody conjugates. Conventional photodynamic therapy (PDT), which combines a photosensitizing agent with the physical energy of non-ionizing light to kill cells, has been less commonly employed for cancer therapy because the current non-targeted photosensitizers are also taken up in normal tissues, thus, causing serious side effects, although the excitation light itself is harmless in the near infrared (NIR) range (FIG. 9). SUMMARY OF THE DISCLOSURE Provided herein are antibody-IR700 molecules and methods of their use for killing a target cell, such as a tumor cell. In particular examples the methods are specific in that non-target cells, such as normal cells, are not killed in significant numbers (such as less than 1% of normal cells are killed), but the target cells are. In particular examples the method includes contacting a cell having a cell surface protein with a therapeutically effective amount of an antibody-IR700 molecule, wherein the antibody (or other specific binding agent) specifically binds to the cell surface protein. Specific non-limiting examples of antibody-IR700 molecules include Panitumumab-IR700, Trastuzumab-IR700, and HuJ591-IR700. The cell is irradiated at a wavelength of 660 to 740 nm, such as 660 to 710 nm (for example, 680 nm) at a dose of at least 1 J cm−2 (such as at least 50 J cm−2). The method also includes contacting the cell with one or more therapeutic agents (such as an anti-cancer agent), for example within about 8 hours after irradiating the cell, thereby killing the cell. Such methods can further include detecting the cell with fluorescence lifetime imaging (FLT), for example about 0 to 48 hours after irradiating the cell, thereby permitting detection of cell killing in real-time. Also provided are methods of detecting cell killing in real-time. Such methods can include contacting a cell comprising a cell surface protein with a therapeutically effective amount of one or more antibody-IR700 molecules as described above, irradiating the cell at a wavelength of 660 to 740 nm and at a dose of at least 30 J cm−2 (such as a dose sufficient to shorten IR700 FLT by at least 25% for example 30 to 50 J cm−2), and detecting the cell with fluorescence lifetime imaging about 0 to 48 hours (such as at least 6 hours) after irradiating the cell, thereby detecting the cell killing in real-time. Any target cell can be killed (and in some examples detected in real-time) with the disclosed antibody-IR700 molecules and methods, for example by using one or more antibodies that binds to one or more proteins on the target cell sur