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

• US 8,874,380 B2
• Filed: 12/09/2011
• Issued: 10/28/2014
• Est. Priority Date: 12/09/2010
• Status: Active Grant

• Current Assignee: Rutgers University
• Original Assignee: Rutgers University
• Inventors: Akudugu, John M., Neti, Venkata S. V. P., Howell, Roger W.
• Primary Examiner(s): Lin, Jerry
• Application Number:
  US13/315,775
• Publication Number:
  US 20120148491A1
• Time in Patent Office: 1,054 Days
• Field of Search: 702/21
• US Class Current: 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 method for predicting the response of an individual patient'"'"'s cells to therapeutic intervention for a condition from which said patient is suffering, said method comprising the steps of:

(a) exposing populations of said cells to increasing concentrations of a candidate therapeutic agent for said condition;

(b) measuring the incorporation of said therapeutic agent in said populations on a cell-by-cell basis;

(c) plotting the number of cells versus the amount of incorporated therapeutic agent, to obtain distribution plots for said populations;

(d) fitting said distribution plots to a probability density function to obtain a distribution curve and the standard deviation, σ

, for each population; and

(e) identifying the optimal concentration of said therapeutic agent for said patient by identifying changes in slope of a plot of σ

as a function of concentration of said therapeutic agent.

2. The method of claim 1, further comprising the step of predicting the surviving fractions of said cell populations employing a simulation that uses the measured incorporation of said therapeutic agent in each cell and a function that represents the probability that a given cell will survive upon incorporating a given amount of the agent.

3. The method of claim 2 wherein said simulation comprises a Monte Carlo simulation.

4. The method of claim 3, wherein said cells are exposed to increasing concentrations of a plurality of candidate therapeutic agents, and the optimal concentration of each therapeutic agent for said patient is identified.

5. The method of claim 4, wherein the Monte Carlo simulation accounts for the amount of each agent in each cell and the results are used to identify the most effective combination of therapeutic agents, and their therapeutically optimal doses in combination.

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

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

8. The method of claim 7, wherein said high-speed technique comprises fluorescence spectroscopy.

9. The method of claim 8, wherein the fluorescence measurement comprises the mean fluorescence intensity (MFI).

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

11. The method of claim 7, wherein the high-speed technique for assaying therapeutic agent incorporation on a cell-by-cell basis comprises fluorescence microscopy or laser scanning microscopy.

12. The method of claim 1, wherein said probability density function is selected from the group of functions consisting of log normal, normal, Weibull and exponential.

13. The method of claim 12, wherein said probability density function is log normal.

14. The method of claim 3, wherein said Monte Carlo simulation comprises flow-cytometry-assisted Monte Carlo simulation.

15. The method of claim 3, wherein said Monte Carlo simulation comprises fluorescence microscope-assisted Monte Carlo simulation or laser scanning microscope-assisted Monte Carlo simulation.

22. The method of claim 2, further comprising determining the surviving fraction of cells, plotting, said surviving fraction versus the amount of incorporated therapeutic agent, and fitting the plot to a probability function selected from the group of functions consisting of exponential and linear-quadratic.

23. A method of high-throughput drug discovery for the treatment of a disease or condition, said method comprising the steps of:

providing a plurality of compounds to be screened for their efficacy in treating said disease or condition; and

predicting the response of a population of cells affected by said disease or condition to therapeutic intervention with every possible combination of said compounds;

wherein said response is predicted using, the method of claim 1.

24. A method of high-throughput drug discovery for treating a type of cancer, said method comprising the steps of:

providing a plurality of compounds to be screened for their efficacy in treating said type of cancer; and

predicting the response of a population of cells affected by said type of cancer to therapeutic intervention with every possible combination of said compounds;

wherein said response is predicted using the method of claim 12.

16. A method for predicting the response of an individual cancer patient'"'"'s cancer cells to therapeutic intervention comprising the steps of:

(a) exposing populations of said cancer cells to increasing, concentrations of a candidate therapeutic agent for said cancer;

(b) measuring by fluorescence spectroscopy the incorporation of said therapeutic agent in said populations on a cell-by-cell basis using a flow cytometer, to provide net mean fluorescence intensity (MFI);

(c) plotting the number of cells versus the net MFI, to obtain a distribution plot for said population;

(d) fitting said distribution plot to a log normal probability density function to obtain a log normal distribution curve and the standard deviation, σ

, for each population;

and

(e) identifying the optimal concentration of said therapeutic agent for said cancer patient from changes in slope of a plot of σ

as a function of concentration of said therapeutic agent.

17. The method of claim 16, further comprising the step of predicting the surviving fractions of said cell populations using a flow-cytometry-assisted Monte Carlo simulation that accounts for the characteristics of said log normal distribution curve.

18. The method of claim 17, wherein said cancer cells are exposed to increasing concentrations of a plurality of candidate therapeutic agents, and the optimal concentration of each therapeutic agent for said cancer patient is identified.

19. The method of claim 18, wherein the simulation results are used to identify the combination of therapeutic agents that affords the highest degree of killing, of said cancer cells.

20. The method of claim 19, wherein the simulation results are used to identify a combination of therapeutic agents that affords a degree of killing of said cancer cells of about 99.99% or greater.

21. The method of claim 19, wherein the simulation results are used to identify one or more therapeutic agents that can be added to a combination of therapeutic agents to facilitate the killing of subpopulations of cells that would otherwise escape killing by said combination.

25. The method of claim 16 wherein said therapeutic agent comprises one or more agents selected from the group consisting of chemotherapeutic agents, radiotherapeutic agents and combinations thereof.