Method for Detecting and Prognosing Pre-Cancerous Cells Using 5, 10, 15, 20-Tetrakis (Carboxyphenyl) Porphine

• US 20100216169A1
• Filed: 03/02/2010
• Published: 08/26/2010
• Est. Priority Date: 11/17/2000
• Status: Active Grant

• Current Assignee: BioAffinity Technologies, Inc.
• Original Assignee: Biomoda, Inc.
• Inventors: Garwin, Jeffrey L.
• Granted Patent: US 7,960,138 B2
• Time in Patent Office: Days
• US Class Current: 435/7.230
• CPC Class Codes: A61K 31/409 having four such rings, e.g. porphine derivatives, bilirubin, biliverdine hemin, hematin A61K31/555
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/574 for cancer

Abstract

Presented is a method of prognosing a patient'"'"'s response to a cancer therapy wherein prior to the therapy contacting a sample of cells from the patient'"'"'s tissue or organ being treated for the cancer with a solution of TCPP to permit binding of the TCPP to components of the abnormal dysplastic or carcinomic cells, if any are present; detecting TCPP fluorescence in the sample, the presence of TCPP fluorescence being indicative that the sample contains dysplastic or carcinomic cells; at intervals during the therapy and subsequent to the therapy performing steps a-c on another sample of cells from the patient'"'"'s tissue or organ being treated for the cancer; and determining if the percentage of abnormal pre-cancerous cells in the samples tested during and subsequent to the therapy are reduced as compared with the sample tested prior to the therapy, the reduction being prognostic of the patients response to the cancer therapy.

Claims

1. A method of prognosing a patient'"'"'s response to a cancer therapy, the method comprising:

a) prior to the therapy, contacting a sample of cells from the patient'"'"'s tissue or organ being treated for the cancer with a solution of TCPP under conditions permitting binding of the TCPP to components of abnormal precancerous or cancerous cells, if any are present;

b) detecting TCPP fluorescence in the sample, the presence of TCPP fluorescence being indicative that the sample contains dysplastic or carcinomic cells;

c) at intervals during the therapy and subsequent to the therapy, performing steps a) to b) on another sample of cells from the patient'"'"'s tissue or organ being treated for the cancer; and

d) determining if the percentage of the abnormal pre-cancerous or cancerous cells in the samples tested during and subsequent to the therapy are reduced as compared with the sample tested prior to the therapy, the reduction being prognostic of the patient'"'"'s response to the cancer therapy.

2. The method of claim 1, further comprising separating the cells by fluorimetric flow cytometry into a population comprising normal cells and a population comprising abnormal precancerous and cancerous cells.

3. A method of prognosing a patient'"'"'s response to a cancer therapy, the method comprising the steps of:

a) prior to the therapy, contacting an unfixed sample of cells from the patient'"'"'s tissue or organ being treated for the cancer with a solution of TCPP under conditions permitting binding of the TCPP to components of abnormal precancerous or cancerous cells, if any are present;

b) detecting TCPP fluorescence in the sample the presence of TCPP fluorescence being indicative that the sample contains dysplastic of carcinomic cells;

c) separating the cells by fluorescence flow cytometry into low or non-fluorescent and high fluorescent populations, the low or non-fluorescent populations being normal or metaplastic, the high-fluorescent population being dysplastic or carcinomic;

d) treating the separated cell populations with a potential cancer therapeutic agent in vitro; and

e) observing the effect of the agent on each of the two cell populations, an observation that the agent is exerting its desired negative effect on the dysplastic or carcinomic cells population and not, or in a reduced amount, on the normal cell population being prognostic of the patient'"'"'s response to the cancer therapy.

4. A kit for detection of dysplastic of carcinomic cells in a sample, the kit comprising TCPP in a container and instructions for its use for detecting of abnormal precancerous or cancerous cells in a sample.

5. The kit of claim 4, further comprising one or more of:

a) components for collecting the sample of cells;

b) components for adhering the sample to a solid support;

c) control cell samples of known cancerous, dysplastic, metaplastic or normal condition; and

d) components for quantitating TCPP fluorescence.

6. A method to determine if a sample of cells contains dysplastic or carcinomic cells, the method comprising the steps of:

contacting the sample with a solution of 5, 10, 15, 20-tetrakis (carboxyphenyl) porphine (TCPP) under conditions permitting binding of the TCPP to components of the dysplastic or carcinomic cells, if any are present, wherein the solution of TCPP comprises the TCPP pre-dissolved in basified alcohol; and

detecting TCPP fluorescence in the sample, the presence of TCPP fluorescence being indicative that the sample contains dysplastic or carcinomic cells.

7. A method of prognosing a patient'"'"'s response to a cancer therapy, the method comprising:

a) prior to the therapy, contacting a sample of cells from the patient'"'"'s tissue or organ being treated for the cancer with a solution of 5, 10, 15, 20-tetrakis (carboxyphenyl) porphine (TCPP) at about room temperature for between about 0.2-3.0 minutes to permit binding of the TCPP to components of abnormal precancerous or cancerous cells, if any are present, wherein the solution of TCPP comprises the TCPP predissolved in basified alcohol;

b) detecting TCPP fluorescence in the sample, the presence of TCPP fluorescence being indicative that the sample contains abnormal precancerous and cancerous cells;

c) at intervals during the therapy and subsequent to the therapy, performing steps a) to b) on another sample of cells from the patient'"'"'s tissue or organ being treated for the cancer; and

d) determining if the percentage of the abnormal pre-cancerous or cancerous cells in the samples tested during and subsequent to the therapy are reduced as compared with the sample tested prior to the therapy, the reduction being prognostic of the patient'"'"'s response to the cancer therapy.

8. The method of claim 7, further comprising separating the cells by fluorimetric flow cytometry into a population comprising normal cells and a population comprising abnormal precancerous and cancerous cells.