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Treatment of circulating tumor cells using an extracorporeal device

a technology of circulating tumor cells and extracorporeal devices, which is applied in the direction of blood circulation devices, cardiovascular disorders, drug compositions, etc., can solve the problems of rare ctcs shed from solid tumors into blood stream that cannot be separated and concentrated in blood effectively in the online, and the current treatment methods for solid tumors are not effective at eliminating ctcs. , to achieve the effect of effective inducing apoptosis, necrosis and/or autophagy

Inactive Publication Date: 2015-12-03
RAKUTEN MEDICAL INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides methods for inducing cell death in circulating tumor cells (CTCs) in a subject through the administration of a therapeutically effective agent and ex vivo irradiation of the subject's blood with a specific light. The method can also involve the use of a targeting agent or a magnetic particle to specifically bind to CTCs for their separation and treatment. The technical effects include reducing the number of CTCs and inducing cell death in cancer treatment.

Problems solved by technology

One of key factors in these kinds of methods is the extensive extinction of excitation light when passing through skins, soft tissues, or organs.
However, the rare CTCs shed from solid tumor into blood stream could not be separated and concentrated from blood effectively in the online extracorporeal device.
Current treatment methods for solid tumors are not effective at eliminating CTCs since they target only the cells of the tumor.

Method used

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  • Treatment of circulating tumor cells using an extracorporeal device
  • Treatment of circulating tumor cells using an extracorporeal device
  • Treatment of circulating tumor cells using an extracorporeal device

Examples

Experimental program
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Effect test

example 1

Using IRDye® 700DX-EGF Probe and Photoactivating Light to Kill Cancer Cells Overexpressing the EGF Receptor

[0086]This example illustrates the application of photodynamic therapy using an IRDye® 700DX-targeting agent (e.g., EGF-700DX) and photoactivating light to kill targeted cells.

[0087]Cancer cells that overexpress the EGF receptor (e.g., A431 cells) were seeded in an in vitro culture system. The cells were incubated with the targeting agent EGF-700DX and exposed to irradiating light at a wavelength of 690 nm and at 8 J / cm2 or 16 J / cm2.

[0088]FIG. 3 shows a reduction in the number of targeted cells after exposure to irradiating light at 8 J / cm2 or 16 J / cm2. FIGS. 3A-E show a reduction in the number of targeted cancer cells after exposure to irradiating light at 8 J / cm2 or 16 J / cm2. FIGS. 3A-C are control data and FIGS. 3D-E show exposure to 8 J / cm2 or 16 J / cm2, respectively. In addition, FIG. 3E shows significant morphology changes in cells treated with EGF-700DX and irradiating li...

example 2

Methods for Inducing Apoptosis, Necrosis or Autophagy in Cancer Cells Using an IRDye® 700DX-EGF Probe and Photoactivating Light

[0094]A study was performed to optimize the conditions for IRDye®700DX-EGF based photodynamic therapy. In the experiment factors (e.g., parameters and variables) that contribute to the level of apoptosis and / or necrosis are tested. In particular, three factors with two conditions for each were evaluated in a 2×2×2 factorial design of experiments. The factors included: A) probe concentration at 0.5 μM and 2.0 μM, B) irradiation of 4 J / cm2 or 16 J / cm2, and C) probe incubation time of 5 minutes or 10 minutes. All possible treatment conditions were tested. Representative images of the treated cells are provided in FIGS. 6A-6D and 7A-7D.

[0095]Briefly, cancer cells that overexpress EGF (e.g., A431 cells) were plated and incubated with the IRDye® 700DX-EGF probe at the test concentration and at a flow rate of 1.36 mL / min. After the incubation, the cells were washed...

example 3

Methods for Inducing Apoptosis, Necrosis or Autophagy in Cancer Cells Using an IRDye® 700DX-EGF Probe or an IRDye® 700DX-Panitumumab Probe

[0100]This example illustrates that the IRDye® 700DX-EGF probe and the IRDye® 700DX-panitumumab probe can induce cell death in A431 cells which are human epithelial cancer cells that overexpress the EGF receptor. EGF is the ligand for the EGF receptor and panitumumab is a fully human monoclonal antibody specific to the receptor.

[0101]A431 cells were plated and incubated with either probe for 5 minutes, 10 minutes or 20 minutes prior to washing and irradiation at 24 J / cm2. The IRDye® 700DX-EGF probe was used at a concentration of 0.5 μM in complete media and the IRDye® 700DX-panitumumab probe was used at a concentration of 0.1 μM. After irradiation, the cells were cultured at 37° C., 5% CO2 for approximately 24 hours. The NucleoCounter® NC-3000™ cell vitality assay (Chemometec, Allerød, Denmark) was performed. Table 1 provides cell numbers. Table 2...

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Abstract

The present invention relates to methods and compositions for eliminating circulating tumor cells (CTCs) in a patient with or without solid tumors. In particular, the ex vivo method utilizes a photodynamic or magnetic targeting moiety that specifically binds to antigens expressed on the cell surface of CTCs.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims priority to U.S. Provisional Patent Application Nos. 62 / 006,790, filed Jun. 2, 2014; 62 / 017,165, filed Jun. 25, 2014; 62 / 066,807, filed Oct. 21, 2014 and 62 / 082,052, filed Nov. 19, 2014, the teachings all of which are hereby incorporated by reference in their entireties for all purposes.BACKGROUND OF THE INVENTION[0002]Tumor metastasis is the process in which cancer cells spread from the site of a primary solid tumor to one or more other organs in the body. The cancer cells, such as circulating tumor cells (CTCs), shed from the solid tumor, translocate to distant tissues through blood and lymphatic vessels, and eventually proliferate and colonize to form metastases.[0003]Solid tumors are treated by therapies including surgery, radiotherapy, chemotherapy and targeted antibody therapy. Photoimmunotherapy using photosensitizing moiety or biomolecule-fluorophore conjugates has been been described as a possible treatmen...

Claims

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Application Information

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IPC IPC(8): A61K41/00A61M1/36
CPCA61K41/0071A61M1/3618A61M1/3683A61M1/3686A61M1/3621A61K51/0446A61K51/0497A61K51/083A61K51/088A61K51/082A61K47/64A61K47/6415A61K47/642A61K47/6849A61K41/10A61P17/00A61P17/06A61P17/10A61P27/02A61P31/04A61P35/00A61P43/00A61P9/00A61P9/10A61K49/0036A61K49/005A61N5/062A61N2005/0658A61K49/0032
Inventor KOVAR, JOY L.WANG, HAN-WEI
Owner RAKUTEN MEDICAL INC
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