KR20160044217A - Composition comprising solubilized photosensitizer for diagnosis and treatment of diseases - Google Patents

Abstract

The present invention relates to a composition for diagnosis and treatment, based on solubilization and, more specifically, to a composition comprising a photosensitizer, glyceryl monooleate, glycerin fatty acid ester, and an emulsifier for diagnosis and treatment. The composition for diagnosis and treatment according to the present invention has excellent solubility by having hydrophilicity, has high metabolism speed, has non-specific accumulation to a normal cell, and is capable of specifically bonding to cancer, thereby maximizing efficiency of diagnosing and treating cancer as a result.

Description

[0001] The present invention relates to a pharmaceutical composition for diagnosing or treating a photosensitizer,

The present invention relates to a composition for diagnosis and treatment, and more particularly to a diagnostic and therapeutic composition comprising a photosensitizer, glyceryl monooleate, a glycerin fatty acid ester and an emulsifier.

Cancer has been the leading cause of death for Koreans over the past 20 years, and the likelihood of cancer to be reached if Koreans survive to the average life expectancy (81 years) is 36.4%, or 37.6% , And women (84 years) are estimated to develop cancer in one out of three (33.3%). The national cancer incidence rate is steadily rising from 1.5% in 1999 to 10% 5.3%, and the importance of early diagnosis of cancer is increasing.

Recently, the incidence of cancer in the gastrointestinal tract has been increasing due to the rapid introduction of Western eating habits, and various studies for early diagnosis have been actively conducted.

Photosensitizer is a substance that has a specific action when it touches light and oxygen. In medical treatment, when it is injected into the human body, it shoots laser beam, and when it is irradiated with oxygen in the body, it generates cell destruction material. These photosensitizers are mainly used for photodynamic therapy (PDT), which is administered to the human body for the purpose of diagnosing and treating cancer in a person suspected of having a malignant tumor. The photodynamic therapy is also used for cancer treatment. Arthritis, and the like. The photosensitizer traces cancer and accumulates at high concentration in cancer tissue. When laser beam of specific wavelength is struck on these sites, it exhibits strong fluorescence and selectively necrotizes cancer tissue.

Photosensitizers have been used in clinical practice due to the application of photofrin and photogem based on hematophorphophyrin derivatives (HpD). HpD-like photosensitizers cause tumor necrosis by inducing vascular damage to the tumor and stopping blood flow. In recent years, research and development on the early diagnosis of cancer using such a photosensitizer as a fluorescent substance has been under way. This is advantageous in that the presence of a target substance can be confirmed by using a simple camera and an integrated device without using expensive diagnostic equipment. Many developments have been made on a diagnostic system in which a bio-target substance and a fluorescent substance are conjugated.

However, photosensitizers such as HpD have a disadvantage in that they have a very long time to be excreted in the body, so that they can not see light for a long time until they are metabolized completely. Most of the sensitizers have hydrophobicity due to their structural characteristics, and they retain hydrophobicity even after bonding with the target material, and thus they have many difficulties in practical application in vivo. In the case of sensitizers with increased hydrophilicity, the nonspecific accumulation in normal cells is reduced, and the faster the release in the body, the shorter the duration of the body is. On the other hand, in order to deliver sufficient amount of the therapeutic agent to the tumor tissue, And it is difficult to accurately diagnose cancer cells because it is difficult to penetrate them. Therefore, there is a need for a new formulation capable of enhancing the fluorescence diagnostic efficiency that can maintain hydrophilicity while enhancing the specificity of the cancer tissue.

Korean Patent Application No. 2010-0020279

The object of the present invention is to provide a composition for diagnosis and treatment on the basis of hydrophilic solubilization. Specifically, the present invention provides a composition for diagnosing and treating cancer by providing hydrophilic properties of a fluorescent diagnostic substance, Which is capable of enhancing the diagnostic efficiency of cancer, and which is also capable of treating cancer. Another object of the present invention is to provide a method for producing a diagnostic and therapeutic composition comprising a novel photosensitizer.

According to one aspect of the present invention, there is provided a diagnostic composition comprising a sensitizer, glyceryl monooleate, a glycerin fatty acid ester, and an emulsifier. Preferably, the sensitizer may be a tetrapyrrole derivative, and more preferably, a target specific peptide or antibody is further bound to enhance diagnostic efficiency. The sensitizer is a tetrapyrrole derivative-polymer in which a chemical bond group (-SH, -OH, -COOH, NH2) is modified so that a peptide or an antibody can be bound thereto. More preferably, the polymer is a polyethyleneglycol , PEG).

The sensitizing agent to which the peptide is bound is pheophorbide A-PEG-Maleimide, chlorin e6-PEG-Maleimide or phthalocyanine-PEG-Maleimide.

The diagnostic composition is characterized by comprising 1 to 20 parts by weight of a sensitizer, 10 to 60 parts by weight of glyceryl monooleate, 29 to 50 parts by weight of glycerin fatty acid ester, and 10 to 20 parts by weight of an emulsifier, Lys monooleate is a monoolein, the glycerin fatty acid ester is Lavrapech WL 1394 and the emulsifier is polysorbate 80.

According to another aspect of the present invention, there is provided a process for preparing a glycerol monooleate comprising: 1) mixing and stirring glyceryl monooleate, a glycerin fatty acid ester and an emulsifier; And 2) adding a sensitizer to the mixed solution and stirring the mixture with an ultrasonic disintegrator.

The mixing temperature in step 1) of the present invention is preferably 30 to 40 ° C, and the mixture is stirred in an ultrasonic mill for 1 to 10 minutes, more preferably for 2 to 5 minutes in step 2).

The composition for diagnosis and treatment according to the present invention is hydrophilic and has excellent solubility, has a rapid metabolic rate, and reduces nonspecific accumulation in normal cells and specifically binds to cancer, thereby maximizing the diagnosis and treatment efficiency of cancer .

Fig. 1 shows the compositions of Examples 1 to 3 and Comparative Examples 1 to 3 of the present invention as a result of visual observation (a), (c) and (e) of Comparative Examples 1 to 3; (b), (d) and (f) are the respective compositions of Examples 1 to 3.

The present invention provides a diagnostic composition comprising a sensitizer, glyceryl monooleate, a glycerin fatty acid ester and an emulsifier.

Photosensitive materials used as photoproducts are broadly classified into porphyrin-based compounds and nonporphyrin-based compounds in the form of free bases or metal complexes.

The photosensitive compound may be selected from the group consisting of porphyrin derivatives; Reduced porphyrins in which at least one pyrrole constituting porphyrin is reduced to pyrroline; Chlorin; Bacteriochlorin; Or porphyrin analogues such as phthalocyanine, naphthalocyanine and the like. In addition, a desired compound can be prepared by introducing a functional group such as a carboxylic acid into the side chain structure by various synthetic methods.

Examples of the non-porphyrin compound include hypericin, rhodamine, rose Bengal, psoralen, phenothiazinium dyes or merocyanine. .

Specifically, fluorescent dyes of the BODIPY series marketed by Molecular Probe Co., for example BODIPY FL; BODIPY TMR STP ester; BODIPY TR-X STP ester; BODIPY 630/650-X STP ester, and BODIPY 650/665-X STP ester, and Alexa series fluorescent dyes commercially available from Molecular Probes Co., for example Alexa Fluor 350 carboxylic acid; Alexa Fluor 430 carboxylic acid; Alexa Fluor 488 carboxylic acid; Alexa Fluor 532 carboxylic acid; Alexa Fluor 546 carboxylic acid; Alexa Fluor 555 carboxylic acid; Alexa Fluor 568 carboxylic acid; Alexa Fluor 594 carboxylic acid; Alexa Fluor 633 carboxylic acid; Alexa Fluor 647 carboxylic acid; Alexa Fluor 660 Carboxylic Acid, and Alexa Fluor 680 Carboxylic Acid. Examples of fluorescent dyes available from Amersham-Pharmacia Biotech include Cy3 NHS ester; Cy 5 NHS ester; Cy5.5 NHS ester, and Cy7 NHS ester. Examples of fluorescent dyes commercially available from VisEn Medical, Inc, (Woburn, Mass.) Include VivoTag 680 and VivoTag 750.

In the present invention, the sensitizer is preferably a tetrapyrrole derivative, more preferably a cyclopentadiene derivative.

 The sensitizer may be further conjugated with a target specific ligand to enhance diagnostic efficiency. Target specific binding ligands are ligands that are specific for target cells, eg, tumors, and enable active targeting. They are DNA, RNA, RNAi, abtamers, PNA, antibodies, folic acid, peptides, Etc., but is not limited thereto, and is preferably a peptide.

The target peptide has a target specificity that enables early diagnosis of disease. For example, in cancer, αvβ3 integrin binding RGD peptide, gastric cancer endothelium binding CGNSNPKSC peptide, vascular cell adhesion molecule-1 targeting (VCAM-1) VHSPNKK peptide, matrix metalloproteinase (MMP-2/9 targeting CTTHWGFTLC and SGKGPRQITAL peptide, urokinase plasminogen activator (uPA) binding SGRSA peptide, urokinase-type plasminogen activator

receptor (uPAR) homing FSRYLWS peptides.

The target peptide may be coupled to a sensitizer to increase the target orientation of the sensitizer and may be modified with various linkers for the binding of the peptide. Preferably, the polymer is bound to a fluorescent material conjugated with a maleimide, more preferably a fluorescent material-polymer-maleimide form.

Polymers can be used for all biocompatible polymers. They are highly stable in vivo and have a high bio-distribution in blood, so they should be able to accumulate continuously in cancer tissues for a sufficient time. Possible examples include dextran ), Chitosan, glycol chitosan, poly-L-lysine and poly-aspartic acid and poly (N-2- ( Poly (divinyl ether-comaleic anhydride) (poly (N-2- (hydroxypropyl) methacrylamide) And synthetic polymers such as poly (styrene-co-maleic anhydride), polyethylene glycol, and the like.

In the present invention, it is preferable that the target specific ligand or the tetrapyrrole derivative-polymer modified with maleimide as the sensitizer to which the peptide can be bound,

More preferably, Pheophorbide A-PEG-Maleimide of Formula 1, Chlorin e6-PEG-Maleimide of Formula 2, or Phthalocyanine-PEG-Maleimide of Formula 3 may be used as a sensitizer. These are fluorescent materials optimized for use in biological imaging due to strong fluorescence intensity and low interference with constituents in vivo. Polyethlene glycol (PEG) can be applied to the polymer. The molecular weight is preferably 2,000 to 6,000.

≪ Formula 1 >

(2)

(3)

The diagnostic composition of the present invention preferably has 1 to 20 parts by weight of sensitizer, 10 to 60 parts by weight of glyceryl monooleate, 29 to 50 parts by weight of glycerin fatty acid ester, and 10 to 20 parts by weight of emulsifier.

Wherein the glyceryl monooleate compound is selected from the group consisting of pheasol, monoolein, monopalmitolane, monomyristolein, monoylidyne, monolucin and monoglycerides semisynthetic from triglycerides of vegetable or animal oils , And monoolein is most preferred.

The glycerin fatty acid ester-based compound may be one or more selected from the group consisting of Labrafac WL 1394, Captex 8000, triacetin, tributyrin, tricaproin, tricaprylin, tricaprin and triolein , And Labrafac WL 1394 is the most preferred.

The emulsifier may be selected from the group consisting of poloxamer (polyoxyethylene-polyoxypropylene copolymer, Pluronic), sorbitan ester (Span), polyoxyethylene sorbitan (Tween) and polyoxyethylene ether (Brij) Of these, polyoxyethylene sorbitol is preferable, and polysorbate 80 is more preferable.

Thus, when the target diagnostic and therapeutic substance is solubilized, the diagnostic substance can be activated and precipitated in an aqueous solution, which may be generated when the target substance is not solubilized, to be diagnosed by binding to a cancer target substance.

In addition, the diagnostic composition is preferably diagnosed and treated for cancer, and the cancer can be selected from the group consisting of digestive, urinary, reproductive, respiratory, circulatory, brain and nervous system cancers.

More specifically, the cancer is selected from the group consisting of lung cancer, non-small cell lung cancer, colon cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, uterine cancer, ovarian cancer, rectal cancer, gastric cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma , Cancer of the uterine cervix, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small bowel cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute From a group consisting of leukemia, lymphocytic lymphoma, bladder cancer, renal or ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma and pituitary adenoma But is not limited thereto.

Such diagnostic compositions may be administered by a route selected from oral, intravenous, intraperitoneal, intramuscular, intracranial, intratumoral, intradermal, transdermal, esophageal, abdominal, But is not limited thereto.

Suitable carriers that can be used for parenteral administration are well known to those skilled in the art. An aqueous vehicle including, but not limited to, sodium chloride injection, Ringer's injection, dextrose injection, dextrose and sodium chloride injection, and lactate-containing Ringer's injection; Water-miscible carriers including but not limited to ethyl alcohol, polyethylene glycol, and polypropylene glycol; And non-aqueous carriers including, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

When such a diagnostic composition is used for tumor diagnosis, the effective amount can be determined by a physician on a case-by-case basis. At this time, the severity of the patient's age, sex, weight, or condition to be treated or diagnosed can be considered. As an example, the polymer-photosensitizer conjugate can be administered at 0.001 [mu] g / kg to 10 mg / kg, based on the equivalent of the photosensitizer and the patient's body weight. In certain instances, the conjugate may be administered at a dose of 0.1 mg / kg to 2 mg / kg, based on the photosensitizer.

In the present invention, the fluorescent substance and the target peptide are chemically conjugated to each other to synthesize a fluorescent diagnostic substance for cancer diagnosis, and the peptide fluorescent substance is separated and used by HPLC. Preparation of specific peptide fluorescent conjugates was described in Preparations 1, 2 and 3.

Best Mode for Carrying Out the Invention Hereinafter, the functions and effects of the present invention will be described in more detail through specific embodiments of the present invention. It is to be understood, however, that these embodiments are merely illustrative of the invention and are not intended to limit the scope of the invention.

< Manufacturing example  1> Chlorine e6 - PEG149 - Maleimide (Scheme 1 and 2)

Chlorin e6-PEG149-Maleimide is prepared as in Scheme 2 after preparation of polymer PEG-Ce6 (Scheme 1).

[Reaction Scheme 1]

Reaction 1. Preparation of Polymer PEG-Ce6

77 mg of NHS (N-Hydroxysuccinimide) and 139 mg of DCC (N, N'-Dicyclohexylcarbodiimide) were dissolved in 50 ml of dimethylsulfoxide (DMSO) as shown in Scheme 1 at room temperature. , P2AM-6) were mixed with 262 mg of Chlorin e6 (molecular weight 596.68) and reacted for 24 hours.

Polyethylene glycol - Chlorine  Junction PEG - Ce6 ) Separation and purification

Hydrophobic chromatography (sephadex LH-20) was performed to selectively isolate and purify only the conjugated product of polyethylene glycol-chlorine e6 prepared in the above Reaction Scheme 1. First, polyethylene glycol-chlorine e6 was dissolved in methanol at a concentration of 5 mg / ml, and then methanol containing polyethylene glycol-chlorine e6 was injected into hydrophobic chromatography (sephadex LH-20), and the ratio of water to methanol The yields were recovered at intervals of 1 minute with a ratio of 1: 9 at a ratio of 5: 5. The product obtained by hydrophobic chromatography (sephadex LH-20) was removed from the methanol using an evaporator and dried by freeze-drying.

[Reaction Scheme 2]

Reaction 2. Preparation of Chlorin e6-PEG149-maleimide

200 mg of the polymer PEG-Ce6 separated by the reaction scheme 1 and the purification process were reacted with 10 mg of NHS (N-Hydroxysuccinimide) and 18 mg of DCC (N, N'-Dicyclohexylcarbodiimide) in anhydrous dimethylsulfoxide (DMSO ) Dissolved in 10 ml of a solvent, and 14 mg of maleimide (molecular weight 217.11) was further mixed and reacted for 24 hours. After the reaction, the reaction product was filtered and then dialyzed with primary distilled water for 3 days using a dialysis membrane (Spectra / Por: mol.wt. cutoff size, 3,500) to remove the dimethylsulfoxide and the catalyst used in the reaction . After dialysis, the final reaction was lyophilized to obtain Chlrorin e6-PEG149-Maleimide.

< Manufacturing example  2> Pheophorbide  A ( PheoA ) - PEG149 - Maleimide (Scheme 3)

Pheophorbide A (PheoA) -PEG149-Maleimide is prepared as in Scheme 3 after preparation of polymer PEG-PheoA.

[Reaction Scheme 3]

139 mg of N-hydroxysuccinimide (NHS) and 77 mg of dicyclohexylcarbodiimide (DCC) were dissolved in 50 ml of dimethylsulfoxide (DMSO) in a solvent , 2 g of polyethylene glycol (PEG) 149 (SunBio, P2AM-6) was mixed with 262 mg of phephorbibe-A (molecular weight 592.68) and reacted for 24 hours. The polymer PEG-PheoA prepared after the reaction was reacted with maleimide in the same manner as in Scheme 2 of Preparation Example 1 and the purification process to obtain Pheophorbide A (PheoA) -PEG149-Maleimide.

< Manufacturing example  3> Phthalocyanine - PEG149 - Maleimide (Scheme 4)

Phthalocyanine-PEG149-Maleimide is prepared as Phthalocyanine-PEG149 and then as shown in Scheme 4.

[Reaction Scheme 4]

Phthalocyanine-PEG149 compound was obtained by the same reaction except that Phthalocyanine (PC) was used instead of Pheophorbide A in Scheme 1 of Preparation Example 1. Then, 10 mg of N-hydroxysuccinimide and 18 mg of dicyclohexylcarbodiimide were dissolved in 10 ml of anhydrous dimethylsulfoxide (DMSO) solvent at room temperature, and 14 mg of maleimide (molecular weight 217.11) was further added thereto The mixture was reacted for 24 hours. After filtration, the reaction product is dialyzed with primary distilled water for 3 days using a dialysis membrane (Spectra / Por, mol. Wt. Cutoff size, 1,000) to remove the dimethylsulfoxide and the catalyst used in the reaction. Phthalocyanine-PEG149-Maleimide was obtained by freeze-drying the final reaction product after dialysis.

< Example  1> Chlorine e6 - PEG149 - Maleimide - peptide &Lt; / RTI &gt;

The N-terminal-SSSPIQGSWTWENGK (Cys) WTWKGIIRLEQ -C terminal of the target specific peptide was conjugated to the Chlorin e6-PEG149-Maleimide compound prepared in Preparation Example 1. The conjugated peptide binding compounds were separated using HPLC (C18 high performance liquid chromatography (HPLC) column and Agilent 1100 series HPLC system (Agilent, Santa Clara, Calif.))

36 mg of the isolated Chlorin e6-PEG149-maleimide-peptide was mixed with 1,000 mg of monoolein, 500 mg of Lavrapech WL 1394 and 180 mg of polysorbate 80 at a temperature of 25 ° C for 1 minute, and the resulting mixed solution was dispersed in an ultrasonic mill After stirring for 2 minutes, a diagnostic composition was prepared.

< Example  2> Pheophorbide  A ( PheoA ) - PEG149 - Maleimide - peptide &Lt; / RTI &gt;

The target specific peptide, N-terminal-SSSPIQGSWTWENGK (Cys) WTWKGIIRLEQ -C terminal was conjugated to the Pheophorbide A (PheoA) -PEG149-Maleimide compound prepared in Preparation Example 2. The conjugated peptide binding compounds were separated using HPLC (C18 high performance liquid chromatography (HPLC) column and Agilent 1100 series HPLC system (Agilent, Santa Clara, Calif.))

36 mg of the separated Pheophorbide A (PheoA) -PEG149-maleimide-peptide was mixed with 1,000 mg of monoolein, 500 mg of Lavalex WL 1394 and 180 mg of polysorbate 80 at 25 ° C for 1 minute, After stirring for 2 minutes with an ultrasonic grinder, a diagnostic composition was prepared.

< Example  3> Phthalocyanine - PEG149 - Maleimide - peptide &Lt; / RTI &gt;

The target specific peptide, N-terminal-SSSPIQGSWTWENGK (Cys) WTWKGIIRLEQ-C terminal was conjugated to the Phthalocyanine-PEG149-Maleimide compound prepared in Preparation Example 3. The conjugated peptide binding compounds were separated using HPLC (C18 high performance liquid chromatography (HPLC) column and Agilent 1100 series HPLC system (Agilent, Santa Clara, Calif.))

36 mg of the separated phthalocyanine-PEG149-Maleimide-peptide was mixed with 1,000 mg of monoolein, 500 mg of Lavalex WL 1394 and 180 mg of polysorbate 80 at 25 ° C for 1 minute, After stirring for a minute, a diagnostic composition was prepared.

PEG149-Maleimide-peptide and Phthalocyanine-PEG149-Maleimide-peptide, prepared in Examples 1 to 3, were added to 1 ml of physiological saline, Similarly, the formulations of Comparative Examples 1 to 3 were prepared.

< Comparative Example  1>

Comparative Example 1 was prepared after adding 2 mg of Chlorin e6-PEG149-Maleimide-peptide prepared in Example 1 to 1 ml of physiological saline.

< Comparative Example  2>

2 mg of the Pheophorbide A (PheoA) -PEG149-Maleimide-peptide prepared in Example 2 was prepared in Comparative Example 2 in the same manner as in Comparative Example 1.

< Comparative Example  3>

2 mg of Phthalocyanine-PEG149-Maleimide-peptide prepared in Example 3 was prepared in the same manner as in Comparative Example 1. [

< Test Example  1>

The compositions prepared in Examples 1 to 3 and the compositions prepared in Comparative Examples 1 to 3 were visually observed. As a result, it was confirmed that Comparative Examples 1 to 3 were not dissolved in saline, whereas Examples 1 to 3 were dissolved to maintain hydrophilicity.

< Test Example  2>

The composition prepared in Examples 1 to 3 (formulation) and the composition prepared in Comparative Examples 1 to 3 (physiological saline) were measured with a spectrophotometer (Agilent 8453 (Agilent, Santa Clara, Calif.)) , And it was confirmed that all of the examples (formulations) were excellent in dissolution compared with the comparative example (physiological saline solution).

Transmittance (T%, 730 nm) Saline solution Formulation Pheophorbide A-PEG-peptide 16.2 ± 0.005 62.7 ± 0.03 Ce6-PEG-peptide 49.3 ± 0.009 62.8 ± 0.02 Phtalocyanine-PEG-peptide 33.8 ± 0.007 65.3 ± 0.01

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (16)

A sensitizer, a glyceryl monooleate, a glycerin fatty acid ester, and an emulsifier. The method according to claim 1,
Wherein the sensitizer is a tetrapyrrole derivative. The method according to claim 1,
Wherein the functional group capable of binding the sensitizer with the peptide is a tetrapyrrole derivative-polymer modified by chemically linkable linker using -SH, -OH, -COOH and NH2. The method of claim 3,
Wherein the sensitizer is a tetrapyrrole derivative-polymer in which maleimide is modified so that the peptide can be bound thereto. 5. The method of claim 4,
Wherein the polymer is polyethyleneglycol (PEG). 6. The method according to any one of claims 3 to 5,
Wherein the sensitizer is Pheophorbide A-PEG-Maleimide, Chlorin e6-PEG-Maleimide or Phthalocyanine-PEG-Maleimide. The method according to claim 1,
Wherein the composition comprises 1 to 20 parts by weight of a sensitizer, 10 to 60 parts by weight of glyceryl monooleate, 29 to 50 parts by weight of a glycerin fatty acid ester, and 10 to 20 parts by weight of an emulsifier. The method according to claim 1,
Wherein the glyceryl monooleate is a monoolein. The method according to claim 1,
Wherein the glycerin fatty acid ester is Lavapec WL 1394. The method according to claim 1,
Wherein the emulsifier is polysorbate 80. The method according to claim 1,
Wherein the diagnosis is an inflammation-related disease caused by excessive production of cancer or blood vessels. 12. The method of claim 11,
Cancer is selected from the group consisting of gastric cancer, esophageal cancer, pancreatic cancer, lung cancer, ovarian cancer, colon cancer, liver cancer, head and neck cancer and gallbladder cancer, said inflammatory diseases being ulcerative colitis, Crohns disease, Wherein the composition is selected from the group consisting of intestinal behcets disease, irritable bowel syndrome and colon polyp. The method according to claim 1,
Wherein the composition is diagnosed by orally administered. 1) mixing and stirring glyceryl monooleate, glycerin fatty acid ester and emulsifier; And 2) adding a sensitizer to the mixed solution and agitating the mixture with an ultrasonic pulverizer. 15. The method of claim 14,
Wherein the mixing temperature in step 1) is from 30 to 40 占 폚, and the mixture is stirred in an ultrasonic mill for 1 to 10 minutes in step 2). 16. The method of claim 15,
Lt; RTI ID = 0.0 &gt; 2 &lt; / RTI &gt; to 5 minutes.

Images