NZ540800A - Methylene blue diagnostic agent and in vivo diagnostic methods for detection of epithelial cancer - Google Patents
Methylene blue diagnostic agent and in vivo diagnostic methods for detection of epithelial cancerInfo
- Publication number
- NZ540800A NZ540800A NZ54080000A NZ54080000A NZ540800A NZ 540800 A NZ540800 A NZ 540800A NZ 54080000 A NZ54080000 A NZ 54080000A NZ 54080000 A NZ54080000 A NZ 54080000A NZ 540800 A NZ540800 A NZ 540800A
- Authority
- NZ
- New Zealand
- Prior art keywords
- methylene blue
- composition
- cancerous
- rinsing
- blue
- Prior art date
Links
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Disclosed is a method for in vivo detection of premalignant epithelial lesions and carcinomas, including the steps of sequentially rinsing the epithelium with dye stain composition which is selectively retained by cancerous and precancerous tissue, wherein the stain composition consists essentially of (a) methylene blue or (b) leucomethylene blue in combination with a pharmaceutically acceptable oxidising agent, and rinsing the epithelium with a rinse composition for removing unretained stain composition.
Description
New Zealand Paient Spedficaiion for Paient Number 540800
5tfo$oo
NEW ZEALAND PATENTS ACT, 1953
Divided out of No. 530244 Itself Divided out of No. 517519 Dated 30 June 2000
COMPLETE SPECIFICATION
DIAGNOSTIC METHODS FOR DETECTION OF EPITHELIAL CANCER
We, ZILA, INC., of 5227 North Seventh Street, Phoenix, Arizona 85014-2800, United States of America, do hereby declare the invention for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statement:
(followed by page la)
INTELLECTUAL PROPERTY OFFICE OF N.Z.
16 APR 2007
RECEIVED
-la-
DIASNOSTIC METHODS FOR DETECTION OF EPITHELIAL CANCER
Described is a novel diagnostic agent for detection of cancerous and precancerous epithelial tissue. Described is a biological stain composition for in vivo detection of cancerous and precancerous tissue, comprising (a)
methylene blue and a pharmaceutically acceptable aqueous solvent, or (b) leucomethylene blue, a pharmaceutically acceptable oxidizing agent, and optionally a pharmaceutically acceptable aqueous solvent.
According to one aspect, the invention relates to a method for in vivo detection of premaliqnant epithelial lesions and carcinomas.
The method including the steps of sequentially:
rinsing the epithelium with stain composition which is selectively retained by cancerous and precancerous tissue, wherein the stain composition consists essentially of (a) methylene blue or (b) leucomethylene blue in combination with a pharmaceutically acceptable oxidising agent, and rinsing the epithelium with a rinse composition for removing unretained stain composition.
INTELLECTUAL PRO PERT OF N.Z
-lb-
In another aspect the invention provides a use of methylene blue in formulating a topical biological stain composition for use for in vivo detection of cancerous and pre-cancerous epithelial tissue.
Described are such diagnostic procedures and agents useful therein which are especially useful for in vivo screening of patients for possible oral cancer as part of routine dentist's or physician's examinations or procedures, such as periodic dental or physical examinations, dental cleaning, etc.
Described are such procedures and compositions useful therein, which use dye stains that are more readily available and/or less expensive and less complicated to synthesize and/or purify than the dyes employed in prior art procedures.
iNTEI-lECTUAL PROPERTY OPPICF OP M.2.
2 I MAR 2007
Ri=rii=n/En
(followed by page 2a)
Described are such in vivo procedures and compositions employing a dye which/ despite prior art teachings otherwise, is sufficiently non-toxic that it can be employed by rinsing the entire oral cavity and/or gargling.
The term "comprising" as used in this specification and claims means "consisting at least in part of"; that is to say when interpreting statements in this specification and claims which include "comprising", the features prefaced by this term in each statement all need to be present but other features can also be present. Related terms such as "comprise" and "comprised" are to be interpreted in similar manner.
In-vivo diagnostic procedures for detecting premalignant epithelial lesions, such as oral lesions and oral carcinomas/ employing dye compositions that are selectively retained by tissues rendered abnormal by dysplasia, hyperplasia, tumoriegnesis and other active surface lesions, are known in the art. For example, procedures employing fluorescein or fluorescein derivatives are disclosed in Chenz, Chinese Journal of Stomatology (27:44-47(1992)) and Pilurin (Stomatologic
\bOEC i
-2a-
(followed by page 3)
(Russian) 72:44-47 (1993)). These procedures involve application of the dye, followed by visual examination under ultraviolet light to detect cancerous/precancerous tissue, which is selectively fluorescent.
Another prior art procedure involves in vivo application by rinsing with toluidine blue 0, followed by normal visual examination to detect any selectively stained tissue. Such procedures are disclosed, for
example, in U.S. Patent 5,372,801 to Tucci, et al. and in U.S. Patent 4,321,251 to Mashberg. Toluidine blue has been used for decades as a histopathological stain for in-vitro use. Through this use it has become known as a metachromatic dye, staining nuclei rich in DNA and RNA a purple to pink color. The inherent deep blue color of toluidine blue 0 is changed to purple or pink when the dye ife bound to nucleic acid or other acidic cellular macromolecules. Of course, this type of staining is dependent on the dye gaining access to internal subcellular structures such as the nucleus. Such access is readily obtained only by "fixing" a tissue sample with formaldehyde or other reagent that disrupts the cellular membrane without destroying general cellular structure.
In contrast to the mechanism involved in. in-vitro use, the staining of oral tissue in vivo by toluidine blue O is due to its ability to penetrate the cell walls and attach to the mitochondria, which retains the dye longer than components of the extracellular matrix.
The Mashberg procedure involves application of the toluidine blue 0 solution as a rinse of the entire oral cavity, with gargling, followed by rinses with water and
acetic acid to remove dye that is not retained by the cancerous or precancerous tissue. The preliminary diagnosis by the Mashberg procedure is then confirmed by direct application of the toluidine blue 0 composition to the suspect site 10-14 days later. The Tucci '801 patent discloses an improved toluidine blue o composition for use according to the general procedure taught by Mashberg.
An in vivo procedure involving use of Lugol's solution (iodine) and toluidine blue 0 was proposed for detecting esophageal cancer synchronous with upper aerodigestive tract cancers in Papazian,
Gastroenterology Clinique et Biologique 9:16-22 (1985).
More recently, Pomerantz U.S. Patent 5,882,627 disclosed a structurally defined class of oxazine and thiazine dyes that are useful in general accordance with the Mashberg diagnostic protocol. However, the Pomerantz '627 patent; expressly excludes methylene blue from this defined class, asserting that it is too toxic for application by oral rinse and/or gargling procedures.
Contrary, however, to the disclosure of the
Pomerantz '627 patent, methylene blue and its ionic derivatives, in the concentrations and quantities employed for in vivo tissue staining, is safely employed as a topically applied selective stain for detecting and/or delineating epithelial cancerous and precancerous tissue.
Suitable compositions of methylene blue for application of the dye to epithelial tissue are prepared by mixing the dye with a suitable pharmaceutically acceptable solvent- Preferably the pH of the methylene blue solution is adjusted with a suitable pharmaceutically acceptable buffer system to yield a final solution that is substantially isotonic and has a pH in the range of approximately 2.5 to 7.0, preferably 4.0-5.0- This can be accomplished by an acetic acid-sodium acetate buffer system. Other suitable buffer systems include citric acid-sodium citrate or mixed acid salt systems such as citric acid-sodium phosphate and the like.
The solvent used to provide the liquid methylene blue dye compositions of the invention is an aqueous solvent. According to the presently preferred embodiment
of the invention, the solvent included a pharmaceutically acceptable, i.e., non-toxic, non-reactive alcohol, e.g.,
ethyl alcohol. Such solvents do not appreciably interfere with the staining mechanism and do not themselves contribute to the reduction of chromo forms of the dye to leuco forms.
Flavoring, stable to the other components of the dye composition, may be added to improve the palatability of the composition if it is to be used as an oral "rinse."
The amount of methylene blue dye in the liquid composition is preferably adjusted to yield a concentration of approximately 1% by weight of the final composition, although higher concentrations can be employed and lower concentrations are at least partiallv effective. At present, I prefer to employ dye compositions containing from about 0.5 to about 3.5% by weight of the methylene blue component.
Also described are compositions for use in accordance with the methods of the invention, in which any leuco form of the dye present in the composition is oxidized to the chromo form, by inclusion of a
INTELLECTUAL PROPERTY OFPICP OF N.2
2 ] MAR 2007 RECEIVED
pharmaceutically acceptable oxidizing agent, in the manner analogous to that disclosed in the Tucci '801 patent.
EXAMPLES
The following examples are presented in order to illustrate practice of the invention to those skilled in the art arid not by way of limitation of the scope thereof, which is defined only by the appended claims.
Example 1
Preparation of Diagnostic Composition
A diagnostic composition is prepared by mixing each of the indicated components in the following proportions (% by weight):
Purified water U.S.P. 83.85
Glacial Acetic Acid U.S.P. 4.61 Sodium Acetate Trihydrate U.S.P. 2.45
SD18 Ethyl Alcohol 7.48
Hydrogen Peroxide 30%, U.S.P. 0.41
IFF Raspberry IC563457 0.20
Methylene Blue 1.00
Example 2 Preparation of Rinse Solution
A rinse solution is prepared by mixing the following components in the indicated proportions (weight %):
98.70 1.00 0.10 0.20
Example 3 Clinical Effectiveness
Purified Water U.S.P. Glacial Acetic Acid U.S.P. Sodium Benzoate U.S.P. IFF Raspberry ICS63457
The clinical effectiveness of the compositions of Examples 1 and 2, is compared to toluidine blue 0
diagnostic control compositions prepared in accordance with the Tucci '801 patent, using the diagnostic protocol disclosed in the Mashberg '251 patent.
Patients are first screened for oral pathology employing the TBO control composition. After identifying potential cancerous or precancerous pathology, all traces of the TBO are removed by repeatedly rinsing the suspect sites with water and the acetic acid rinse of Example 2.
Those patients exhibiting oral pathology are then used as test subjects for the methylene blue diagnostic composition of Example 1- 2-3 cc of the methylene blue composition is applied by painting the pathologic mucosal surface, followed by rinsing with the rinse mixture and water to remove excess methylene blue composition.
Histological examination of tissue from the areas stained by the methylene blue diagnostic composition of Example 1 confirms that the methylene blue composition is at least as effective as toluidine blue o in identifying and delineating cancerous and precancerous epithelial tissue.
-10-Bxample 4
The procedures of Example 3 are repeated, except that the test and control compositions are applied to the oral mucosa by rinsing, with gargling, instead of by direct application to the locus of the suspect sites. Equivalent results are obtained.
Claims (4)
1. A method for in vivo detection of premalignant epithelial lesions and carcinomas, including the steps of sequentially: rinsing the epithelium with stain composition which is selectively retained by cancerous and precancerous tissue, wherein the stain composition consists essentially of (a) methylene blue or (b) leucomethylene blue in combination with a pharmaceutically acceptable oxidising agent, and rinsing the epithelium with a rinse composition for removing unretained stain composition.
2. A use of methylene blue in formulating a topical biological stain composition for use for in vivo detection of cancerous and pre-cancerous epithelial tissue.
3. A method according to claim 1 substantially as herein described with reference to any example thereof.
4. A use according to claim 2 substantially as herein described with reference to any example thereof. INTELLECTUAL PROPERTY OFFIOF OF \ 2 2 t MAR 2007 REOFlUPn
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
NZ54080000A NZ540800A (en) | 2000-06-30 | 2000-06-30 | Methylene blue diagnostic agent and in vivo diagnostic methods for detection of epithelial cancer |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
NZ54080000A NZ540800A (en) | 2000-06-30 | 2000-06-30 | Methylene blue diagnostic agent and in vivo diagnostic methods for detection of epithelial cancer |
Publications (1)
Publication Number | Publication Date |
---|---|
NZ540800A true NZ540800A (en) | 2007-06-29 |
Family
ID=38190679
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
NZ54080000A NZ540800A (en) | 2000-06-30 | 2000-06-30 | Methylene blue diagnostic agent and in vivo diagnostic methods for detection of epithelial cancer |
Country Status (1)
Country | Link |
---|---|
NZ (1) | NZ540800A (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104483473A (en) * | 2014-12-31 | 2015-04-01 | 广州市微米生物科技有限公司 | Detection reagent for epithelial tissue cancer cells and preparing and detecting method |
CN104483472A (en) * | 2014-12-19 | 2015-04-01 | 武汉大学 | Diagnostic reagent for cervical cancer cell detection and preparation method thereof |
-
2000
- 2000-06-30 NZ NZ54080000A patent/NZ540800A/en unknown
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104483472A (en) * | 2014-12-19 | 2015-04-01 | 武汉大学 | Diagnostic reagent for cervical cancer cell detection and preparation method thereof |
CN104483473A (en) * | 2014-12-31 | 2015-04-01 | 广州市微米生物科技有限公司 | Detection reagent for epithelial tissue cancer cells and preparing and detecting method |
CN104483473B (en) * | 2014-12-31 | 2016-03-02 | 广州市微米生物科技有限公司 | For detection reagent and the preparation and determination methods method of epithelial neoplasms cell |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP0814847B1 (en) | Methods and compositions for in-vivo detection of oral cancers and precancerous conditions | |
Longin et al. | Comparison of anti-fading agents used in fluorescence microscopy: image analysis and laser confocal microscopy study. | |
WO2013097771A1 (en) | Composition of detection agents for epithelial tumour cells and preparation method therefor | |
EP0959906A1 (en) | Delta 1,6 bicyclo [4,4,0] functional dyes for contrast enhancement in optical imaging | |
AU781042B2 (en) | Methylene blue diagnostic agent and diagnostic methods for detection of epithelial cancer | |
CN108844937A (en) | A kind of water soluble fluorescence silicon point and preparation method thereof and application | |
JP2018162986A (en) | Coloring method, coloring material and coloring kit | |
NZ540800A (en) | Methylene blue diagnostic agent and in vivo diagnostic methods for detection of epithelial cancer | |
AU784558B2 (en) | Rhodamine diagnostic agent and diagnostic methods for detections of epithelial cancer | |
RU2178565C2 (en) | Methods and compositions to detect oral malignant tumors and precancerous states in vivo | |
US6967015B1 (en) | Diagnostic method for detecting dysplastic epithelial tissue | |
CN113358615A (en) | Application of melane and fluorescein sodium double staining method in living cell imaging | |
US20080069776A1 (en) | Histofluorescent stain for endoscopy | |
ES2248061T3 (en) | IN VIVO STATION COMPOUNDS AND METHODS OF USE TO IDENTIFY DISPLASIC FABRICS. | |
CN117804876B (en) | Folic acid receptor mediated epithelial tissue cell staining solution and preparation method thereof | |
EP3373009B1 (en) | Fluorescent compounds for the detection of malignant melanoma | |
WO2023181060A1 (en) | Synthesis of toluidine blue o and a kit for mucosal application | |
AU2004324049A1 (en) | Light-directed method for detecting and aiding further evaluation of abnormal mucosal tissue |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
RENW | Renewal (renewal fees accepted) | ||
PSEA | Patent sealed |