Method of overcoming therapeutic limitations of nonuniform distribution of radiopharmaceuticals and chemotherapy drugs

• US 9,804,167 B2
• Filed: 10/28/2014
• Issued: 10/31/2017
• Est. Priority Date: 12/09/2010
• Status: Active Grant

• Current Assignee: Rutgers University
• Original Assignee: Rutgers University
• Inventors: Akudugu, John M., Neti, Venkata S., Howell, Roger W.
• Primary Examiner(s): Lin, Jerry
• Application Number:
  US14/526,041
• Publication Number:
  US 20150265730A1
• Time in Patent Office: 1,099 Days
• Field of Search: 702 21
• CPC Class Codes: A61K 31/704 attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin digitoxin A61K31/7048
• CPC Class Codes: A61K 51/0402 carboxylic acid carriers, fatty acids amino acids A61K51/0406
• CPC Class Codes: A61K 51/1051 the tumor cell being from breast, e.g. the antibody being herceptin
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5011 for testing antineoplastic activity
• CPC Class Codes: G01N 33/60 involving radioactive labelled substances

Abstract

Disclosed is a method for predicting the optimal amounts of radiopharmaceutical and/or chemotherapy agents to administer to a patient, by determining the level of saturation of the therapeutic agents in the patient'"'"'s cells. The method comprises measuring cellular incorporation of the candidate therapeutic agents in a target cell population on a cell-by-cell basis. The method is able to identify an optimal cocktail of therapeutic agents for treatment of a disease. A method of high-throughput drug discovery incorporating this method, and a 2-stage targeting method of treating a disease using this method are also disclosed.

Claims

1. A 2-stage targeting method of treating a disease or condition for a patient in need thereof, said method comprising:

(1) identifying and providing a plurality of candidate cell-specific agents relevant to the disease or condition of said patient, wherein said cell-specific agents are two-stage agents comprising;

(a) one or more Stage 1 targeting agents which target diseased or affected cells of said patient; and

(b) one or more Stage 2 binding agents which bind to said Stage 1 targeting agents and are capable of carrying at least one additional agent selected from the group consisting of toxins, radionuclides and fluorochromes, wherein each Stage 2 binding agent can only bind to a single corresponding Stage 1 targeting agent;

(2) injecting said patient with a cocktail of Stage 1 targeting agents via a route appropriate to said disease or condition, and allowing sufficient time for maximum uptake by said diseased or affected cells and substantial clearance of unbound Stage 1 targeting agents;

(3) withdrawing a sample of said patient'"'"'s diseased or affected cells loaded with Stage 1 targeting agents;

(4) treating said sample of said cells in vitro with a cocktail of Stage 2 binding agents, wherein each Stage 2 binding agent in the cocktail carries at least one additional agent which is a unique fluorochrome, and wherein said Stage 2 binding agents bind to said Stage 1 targeting agents loaded into said cells;

(5) quantifying the amount of each Stage 2 binding agent binding to each diseased or affected cell using fluorescence spectroscopy to provide quantitative measured binding data, and plotting the number of cells versus the amount of bound Stage 2 binding agent to obtain distribution plots for said cells;

(6) predicting the response of said diseased or affected cells for every possible combination of Stage 1 targeting agents and Stage 2 binding agents using a Monte Carlo simulation that uses (i) said measured binding in each cell of said Stage 2 binding agent carrying a fluorochrome as the additional agent, and (ii) a function that represents the probability that a given cell will survive upon binding a given amount of an armed Stage 2 binding agent carrying a toxin and/or radionuclide as the additional agent; and

(a) identifying the predicted optimal combination of said armed Stage 2 binding agents when said cells are exposed to increasing concentrations of a plurality of armed Stage 2 binding agents, (b) identifying the predicted optimal concentration of each armed Stage 2 binding agent for said patient, and (c) said Monte Carlo simulation accounts for the predicted amount of each armed Stage 2 binding agent in each cell;

(7) arming each Stage 2 binding agent of said predicted optimal combination with one or more therapeutic agents selected from the group consisting of toxins, radionuclides, and combinations of two or more thereof, to form an armed cocktail;

(8) optionally, repeating step (2); and

(9) injecting said armed cocktail into said patient.

2. The method of claim 1, further comprising repeating steps (3) through (6) with healthy cells of said patient to assess uptake of said Stage 1 targeting agents and Stage 2 binding agents in each healthy cell.

3. The method of claim 1, wherein said individual patient'"'"'s cells comprise cancer cells.

4. The method of claim 1, wherein said incorporation of said therapeutic agent is measured using a high-speed technique.

5. The method of claim 4, wherein said high-speed technique comprises fluorescence spectroscopy.

6. The method of claim 5, wherein the fluorescence measurement comprises the mean fluorescence intensity (MFI).

7. The method of claim 4, wherein the high-speed technique for assaying therapeutic agent incorporation on a cell-by-cell basis comprises flow cytometry.

8. The method of claim 3, wherein the high-speed technique for assaying therapeutic agent incorporation on a cell-by-cell basis comprises fluorescence microscopy or laser scanning microscopy.

9. The method of claim 1, wherein said Monte Carlo simulation comprises flow-cytometry-assisted Monte Carlo simulation.

10. The method of claim 1, wherein said Monte Carlo simulation comprises fluorescence microscope-assisted Monte Carlo simulation or laser scanning microscope-assisted Monte Carlo simulation.