Treatment protocol

a treatment protocol and technology of a treatment protocol, applied in the field of diagnostic tests, can solve the problems of affecting and requiring time-consuming steps. to achieve the effect of enhancing the antiangiogenic effect of primary drugs

Inactive Publication Date: 2013-01-10
WEISENTHAL LARRY MARK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This method enables efficient detection and quantification of endothelial cells, indicating conditions such as diabetes and cardiovascular issues, and can assess therapeutic effects, while also potentiating antiangiogenic drug effects with nontoxic levels of DMSO and ethanol.

Problems solved by technology

Detecting such cells in the circulatory system is, however, problematic because of interference from other cells, such as platelets.
Methods that involve separation of cells by type, or antibody labeling such as the use of antibodies directed to CD31, require time-consuming steps and often require expensive equipment.
Direct antineoplastic drugs result in the death of the tumor cells themselves, rather than the endothelial cells.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Detection of Endothelial Cells

[0052]Using the procedures set forth in Preparation A, a pancreatic carcinoid tumor was visualized after 96 hours in suspension culture in the presence and absence of 2.5 mg / ml bevacizumab. The cultures were stained with the above-described Fast Green / H&E stain / counterstain. FIG. 1B shows the results in a control culture that has not been treated with bevacizumab. It is apparent that the endothelial cells are barely visible. FIG. 1A shows the results of the control culture (not treated with bevacizumab) when CD31 staining is used to identify the endothelial cells. Again, they are not particularly conspicuous. FIG. 1C shows the results when the culture has been treated with bevacizumab, resulting in endothelial cell death. The appearance of the endothelial cells is quite striking. The characteristic “blueberry pancake” appearance described in the above-referenced WO 2007 / 075440 publication is observed.

example 2

Comparison of Various Chemotherapeutic Agents

[0053]The cultures of Example 1 were treated with various candidate chemotherapeutic drugs in comparison to bevacizumab at 2.5 mg / ml and stained as described in Example 1. The various drugs were rated by scoring on a 0+ to 5+ scale for the anti-microvascular effects, with 5+ being the highest possible score. As shown in FIGS. 2A-2H, gefitinib scored only 0; sunitinib scored 1+, sorafenib scored 3+ and both erlotinib and bevacizumab scored 4+. The “nib” drugs are kinase inhibiting drugs.

[0054]Further results are tabulated in Table 1 where slides were scored for anti-microvascular effects as described above and also for direct antitumor effects as described on the Weisenthal.org website. For this determination, the results provided in Table 1 are 10× multiples of the scoring system of the previous paragraph. The table also shows the concentration of each drug employed. Bevacizumab was dispensed both in a vehicle containing 0.9% NaCl and in ...

example 3

Synergistic Effects of Combinations

[0058]A specimen of poorly differentiated breast cancer was used in the procedures set forth in the previous examples and tested with various drugs and drug combinations as described above. FIGS. 3A-3H show the results. FIG. 3A shows the appearance after staining with Fast Green / H&E on a control culture. FIGS. 3B, 3C and 3D show the appearance when treated with erlotinib, imatinib and bevacizumab alone. None appeared particularly effective, although bevacizumab was scored as +1. Combinations of bevacizumab with either erlotinib (FIG. 3E) or imatinib (FIG. 3F) showed much better results. FIGS. 3G and 3H are simply greater magnifications of the results shown in FIG. 3F.

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Abstract

Combination treatments with an antiangiogenesis agent and non-toxic blood levels of ethanol and / or DMSO are disclosed.

Description

CROSS-REFERENCE TO RELATED APPLICATION[0001]The present application is a divisional of U.S. application Ser. No. 12 / 994,105 having an international filing date of 22 May 2009, which is the national phase of PCT application PCT / US2009 / 045062 having an international filing date of 22 May 2009, which claims benefit under 35 U.S.C. §119(e) to provisional U.S. application No. 61 / 055,413 filed 22 May 2008. The contents of the above patent applications are incorporated by reference herein in their entirety.TECHNICAL FIELD[0002]The invention relates to diagnostic tests useful in evaluative screening of subjects for general wellbeing. More particularly, the invention is directed to an inexpensive and convenient assay for circulating endothelial cells and for detection of endothelial cells in general. In addition, the invention relates to an improved protocol for treatment of conditions characterized by unwanted neovasculature, including neoplastic conditions.BACKGROUND ART[0003]It is general...

Claims

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Application Information

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Patent Type & AuthorityApplications(United States)
IPC IPC(8): A61K31/045A61K31/10A61P9/00A61K39/395
CPCA61K39/3955A61K45/06G01N33/5091G01N33/52G01N2510/00A61K2300/00A61P9/00
InventorWEISENTHAL, LARRY MARK
OwnerWEISENTHAL LARRY MARK