The present invention relates to the field of topical formulation to diagnose cancerous and pre-cancerous oral lesions. More specifically the present invention relates to a formulation which can easily and effectively penetrate through cancerous cells than normal cells which can be easily detected through naked
eyes.
BACKGROUND ART
[0002] India has the one of the most numbers of tobacco consumers in the world which makes India among the top three of the countries having most numbers of oral carcinomas. India itself accounts for the one third of the total oral cancer cases across the world! According to a study there were 119,992 new oral cancer cases within the country and 72,616 deaths due to the same in the year 2018. Due to these drastic figures the country required a diagnostic kit which is Economic, efficient, painless and non-invasive in nature.
[0003] Cancer has been one of the leading causes of death worldwide also. Different types of cancers are difficult to effectively get diagnosed and treat. Accordingly, there is a need in the art for new and effective compositions and methods for assessing and treating various cancers.
[0004] Various tedious approaches have been conducted to detect cancer including detection of the level of protein bio markers, using formulations comprising anti-epidermal growth factor receptor (EGFR) antibody-drug conjugate (ADC), and many more.
[0005] JP5812545B2 discloses In a preferred embodiment of the present invention, for example, the following is provided A method of detecting lung cancer in an individual, the method comprising detecting the level of TFPI in a sample from the individual, wherein a high level of TFPI indicates that the individual has lung cancer .

[0006] US7960512B2 discloses molecules, particularly polypeptides, more particularly immunoglobulins (e.g., antibodies), comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, which variant Fc region binds FcyRIIIA and/or FcyRIIA with a greater affinity, relative to a comparable molecule comprising the wild-type Fc region. The molecules of the invention are particularly useful in preventing, treating, or ameliorating one or more symptoms associated with a disease, disorder, or infection. The molecules of the invention are particularly useful for the treatment or prevention of a disease or disorder where an enhanced efficacy of effector cell function (e.g., ADCC) mediated by FcyR is desired, e.g., cancer, infectious disease, and in enhancing the therapeutic efficacy of therapeutic antibodies the effect of which is mediated by ADCC.
[0007] In order to overcome the aforementioned drawbacks, there is a need in the art to provide a topical formulation which can detect and differentiate between normal cells and cancerous cells/pre-cancerous cells.
OBJECTS OF THE INVENTION
[0008] An object of the present invention is to develop a topical formulation for the mass screening of the oral cancer for at-risk population.
[0009] Another object of the present invention is to differentiate between the normal and cancerous cells by the change in color to blue or purple of the cancerous cells which can be visualized by the naked eyes.
[0010] Another object of the present invention to increase flexibility time for treatment, patient friendliness, efficiency of detection therapy, and/or patient safety.
[0011] Another object of the present invention is to improve cost-effectiveness and convenience of use.

[0012] The foregoing and other objects of the present invention will become readily apparent upon further review of the following detailed description of the embodiments as illustrated in the accompanying drawings.
SUMMARY OF THE INVENTION
[0013] The present invention relates to the field of topical formulation to diagnose cancerous and pre-cancerous oral lesions. More specifically the present invention relates to a formulation which can easily and effectively penetrate through cancerous cells than normal cells which can be easily detected through naked
eyes.
[0014] A formulation, comprising: i)Tolonium Chloride in the range of 0.5 - 1% w/w; ii) Hydrogen peroxide or Ascorbic acid in the range of 0.05 - 0.2% w/w; iii) Acetic acid or citric acid in the range of 4 - 5% w/w; iv) Sodium acetate or Citrate in the range of 0.1 - 2% w/w; v) Ethanol in the range of 4 - 6% w/v; vi) Povidone solution or Lugol's iodine in the range of 0.1 - 3% w/w; v) Acetone in the range of 0.01 - 0.5% w/v; and vi) Flavoring agent in the range of 0.01 - 0.2% vii) water.
[0015] While the invention has been described and shown with reference to the preferred embodiment, it will be apparent that variations might be possible that would fall within the scope of the present invention.
DETAILED DESCRIPTION OF THE INVENTION
[0016] While the present invention is described herein by way of example using embodiments and illustrative drawings, those skilled in the art will recognize that the invention is not limited to the embodiments of drawing or drawings described and are not intended to represent the scale of the various components. Further, some components that may form a part of the invention may not be illustrated in certain figures, for ease of illustration, and such omissions do not limit the embodiments outlined in any way. It should be understood that the drawings and

the detailed description thereto are not intended to limit the invention to the particular form disclosed, but on the contrary, the invention is to cover all modifications, equivalents, and alternatives falling within the scope of the present invention as defined by the appended claim.
[0017] As used throughout this description, the word "may" be used in a permissive sense (i.e., meaning having the potential to), rather than the mandatory sense, (i.e., meaning must). Further, the words "a" or "an" mean "at least one" and the word "plurality" means "one or more" unless otherwise mentioned. Furthermore, the terminology and phraseology used herein are solely used for descriptive purposes and should not be construed as limiting in scope. Language such as "including," "comprising," "having," "containing," or "involving," and variations thereof, is intended to be broad and encompass the subject matter listed thereafter, equivalents, and additional subject matter not recited, and is not intended to exclude other additives, components, integers, or steps. Likewise, the term "comprising" is considered synonymous with the terms "including" or "containing" for applicable legal purposes. Any discussion of documents, acts, materials, devices, articles, and the like are included in the specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention.
[0018] In this disclosure, whenever a composition or an element or a group of elements is preceded with the transitional phrase "comprising", it is understood that we also contemplate the same composition, element, or group of elements with transitional phrases "consisting of, "consisting", "selected from the group of consisting of, "including", or "is" preceding the recitation of the composition, element or group of elements and vice versa.
[0019] The present invention is described hereinafter by various embodiments with reference to the accompanying drawing, wherein reference numerals used in the accompanying drawing correspond to the like elements throughout the

description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiment set forth herein. Rather, the embodiment is provided so that this disclosure will be thorough and complete and will fully convey the scope of the invention to those skilled in the art. In the following detailed description, numeric values and ranges are provided for various aspects of the implementations described. These values and ranges are to be treated as examples only and are not intended to limit the scope of the claims. In addition, several materials are identified as suitable for various facets of the implementations. These materials are to be treated as exemplary and are not intended to limit the scope of the invention.
[0020] The present invention pertains to the formulation incorporated with iodophor that helps to differentiate between the normal and cancerous cells.
[0021] The formulation comprises of Tolonium Chloride (0.5 - 1%), Hydrogen peroxide/Ascorbic acid (0.05 - 0.2%), acetic acid (4 - 5%)/citric acid (0.5 -1.5%), Sodium acetate/ Citrate (0.1 - 2%), ethanol (4 - 6%), Povidone solution/Lugol's Iodide (0.1 - 3%), acetone (0.01 - 0.5%), Flavouring agent (0.01 - 0.2%) and water (QS).
[0022] The iodine solution used in the present invention can be selected from one or more forms of iodine including iodophors and commercially available iodine formulation. The formulation can be used as a solo or in combinations in different suitable ratios in the formulation to detect oral cancer and may also be used to cure/treat cancer.
[0023] In the povidone solution, K-30, K-90, K-15 can be used to make required percentage (5-10%) of iodine solution that can be used in the formulation for the best efficacy. Lugol's iodine can be used directly in the formulation otherwise can be separately made and use in the final formulation.
[0024] The flavoring agent included in the present formulation may include but not limited to, menthol, chocolate, butter scotch, pineapple, vanilla other

commercially available flavoring agent can also be used without losing any efficacy of the formulation.
[0025] The steps involved in usage of the formulation of the present invention for the detection of oral cancer includes: i) Rinsing mouth of the patient or user thoroughly with water ii) Rinsing mouth with 1% acetic acid solution for 30 seconds, iii) With the help of cotton applicator, the formulation is applied to the suspected area and the near-by tissues and kept for 30 to 60 seconds, iv) The mouth of patient is again rinsed with 1% acetic acid solution for about 30 seconds. v) Mouth is rinsed with water vi) Visualization of the purple or deep blue color of the cells can be done through naked eyes.
[0026] The formulation has the good permeability towards the cancer cells more than normal cells which can differentiate between cancerous/pre-cancerous from normal cells that can be observed through naked eyes as blue/purple color stain. It does so on the basis of the excess amount of the nucleic acid present in the cancerous and pre-cancerous cells.
[0027] Tolonium chloride is a vital, acidic cell staining metachromasic dye which has affinity towards anions such as sulphates, carboxylates and phosphates. An oral dysplasia or carcinoma has abnormally high number of nucleic acid due to which cationic groups of the dye are attracted towards the nucleic acid and stain them. Due to metachromasia there is shift in the absorption spectra of the dye which makes it appear purple. The presence of the purple colour after applying formulation indicates the Prescence of the cancerous or pre-cancerous tissue. Accordingly, the formulation can stain any given acidic tissues in the body like mast cells and endocrine cell granules and as it stands the tolonium chloride based iodophor containing formulation, can also differentiate between gastric malignant and benign ulcers.
[0028] The dye can also be activated to produce singlet oxygen in the 810nm to 860nm of the infrared region. This increases the ROS within the cell and causes apoptosis which can even kill microbes and possibly cancer cells as well. Due to

these properties the formulation can be used in PDT (photodynamic therapy) for neutralizing microbes and treating periodontitis.
[0029] The solution has the components like iodophors which will increase the permeability of the tolonium chloride into the carcinoma cells due to their ability to open small pores in the cell membranes. Along with these, other components include a preservative for increasing the shelf life of the formulation like sodium benzoate and ascorbic acid. Also, buffering agents are added to increase the stability of tolonium chloride in the formulation.
RESULTS
[0030] Different formulations were tested for the results:
[0031] Formulation I: Tolonium Chloride (0.5 - 1%), Hydrogen peroxide (0.05
- 0.2%) and/or Ascorbic acid (0.05 - 0.2%), favouring agent (0.1 -0.3%), Acetic acid (4% - 5%), ethanol (4.5 - 6%), Povidone solution (0.5 - 2%), Acetone (0.01
- 0.05%) and water (QS).
[0032] Formulation II: Tolonium Chloride (0.5 - 1%), Hydrogen peroxide (0.05
- 0.2%) and/or Ascorbic acid (0.05 - 0.2%), favouring agent (0.1 -0.3%), Acetic
acid (4% - 5%), ethanol (4.5 - 6%), Povidone solution (0.5 - 2%), and water
(QS).
[0033] Formulation III: Tolonium Chloride (0.5 - 1%), Hydrogen peroxide (0.05 - 0.2%) and/or Ascorbic acid (0.05 - 0.2%), favouring agent (0.1 -0.3%), Acetic acid (4% - 5%), ethanol (4.5 - 6%), Acetone (0.01 - 0.05%) and water (QS).
[0034] Formulation IV: Tolonium Chloride (0.5 - 1%), Hydrogen peroxide (0.05 - 0.2%) and/or Ascorbic acid (0.05 - 0.2%), favouring agent (0.1 -0.3%), Acetic acid (4% - 5%), Sodium Acetate (0.1 - 0.2%), ethanol (4.5 - 6%), Povidone solution (0.5 - 2%) and water (QS).

[0035] Formulation V: Tolonium Chloride (0.5 - 1%), Hydrogen peroxide (0.05
- 0.2%) and/or Ascorbic acid (0.05 - 0.2%), favouring agent (0.1 -0.3%), citric
acid (0.5 - 1.5%), Sodium Citrate (0.1 - 0.2%), ethanol (4.5 - 6%), Povidone
solution (0.5 - 2%) and water (QS).
[0036] Formulation VI: Tolonium Chloride (0.5 - 1%), Hydrogen peroxide (0.05 - 0.2%) and/or Ascorbic acid (0.05 - 0.2%), favouring agent (0.1 -0.3%) (0.1 -0.3%), citric acid (0.5 - 1.5%), Sodium Citrate (0.1 - 0.2%), ethanol (4.5 -6%), Povidone solution (0.5 - 2%) and water (QS).
[0037] Formulation VII: Tolonium Chloride (0.5 - 1%), Hydrogen peroxide (0.05 - 0.2%) and/or Ascorbic acid (0.05 - 0.2%), favouring agent (0.1 -0.3%), citric acid (0.5 - 1.5%), Sodium Citrate (0.1 - 0.2%), ethanol (4.5 - 6%), Lugol's Iodide (0.1 - 3%) and water (QS).
[0038] Formulation VII: Tolonium Chloride (0.5 - 1%), Hydrogen peroxide (0.05
- 0.2%) and/or Ascorbic acid (0.05 - 0.2%), favouring agent (0.1 -0.3%), Acetic
acid (4% - 5%), ethanol (4.5 - 6%), Polyvinylpyrrolidone (0.5 - 4%), Acetone
(0.01 - 0.05%) and water (QS).
[0039] Formulation VIII: Tolonium Chloride (0.5 - 1%), Hydrogen peroxide (0.05 - 0.2%) and/or Ascorbic acid (0.05 - 0.2%), favouring agent (0.1 -0.3%), Acetic acid (4% - 5%), ethanol (4.5 - 6%), Polyvinylpyrrolidone (0.5 - 4%), and water.
[0040] ADVANTAGES and APPLICATIONS
1. Use as oral cancer screening and diagnosis.
2. Can be used in periodontitis treatment.
3. Can be used to identify the uterine cervical carcinoma.
4. Use as a photosensitizer in various PDT based therapy.
5. Use as a stain for staining the acidic tissues and cells.
[0041] In an alternative embodiment, the formulation of the present invention can

be used to treat multidrug resistant microbes at normal conditions and/or in presence of Infrared Radiation stimulation of different wavelengths.
[0042] Further, the operations need not be performed in the disclosed order, although in some examples, an order may be preferred. Also, not all functions need to be performed to achieve the desired advantages of the disclosed system and method, and therefore not all functions are required.
[0043] Various modifications to these embodiments are apparent to those skilled in the art from the description and the accompanying drawings. The principles associated with the various embodiments described herein may be applied to other embodiments. Therefore, the description is not intended to be limited to the 5 embodiments shown along with the accompanying drawings but is to be providing the broadest scope consistent with the principles and the novel and inventive features disclosed or suggested herein. Accordingly, the invention is anticipated to hold on to all other such alternatives, modifications, and variations that fall within the scope of the present invention and appended claims.

CLAIMS
I/We Claim:

1) A formulation, comprising:
i) Tolonium Chloride in the range of 0.5 - 1% w/w;
ii) Hydrogen peroxide or Ascorbic acid in the range of 0.05 - 0.2% w/w;
iii) Acetic acid or citric acid in the range of 0.5 - 5% w/w;
iv) Sodium acetate or Citrate in the range of 0.1 - 2% w/w;
v) Ethanol in the range of 4 - 6% w/v;
vi) Povidone solution in the range of 0.1 - 3% w/w;
vii) Acetone in the range of 0.01 - 0.5% w/v; and
viii)Flavouring agent in the range of 0.01 - 0.2%.
2) The formulation as claimed in claim 1, wherein said formulation is prepared by
incorporating water.
3) The formulation as claimed in claim 1, wherein the said formulation works best
with an iodophor like povidone, lugol,s iodine etc.
4) The formulation as claimed in claim 1, wherein said flavoring agent may
include but not limited to food flavoring agents
5) The formulation as claimed in claim 1, wherein said formulation may be used
as an effective agent to detect the presence of cancerous and pre-cancerous cells.