JP2005145921A - Fluorescent contrast medium for diagnosis and method for fluorescence imaging diagnosis - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a fluorescent contrast medium for diagnosis of cancer or tumor having low toxicity and excellent water solubility, emitting fluorescence in a region transmittable through biological tissues and providing specific imaging of the tumor, cancer and/or blood vessels, and to provide a method for fluorescence imaging diagnosis. <P>SOLUTION: The fluorescent contrast medium for diagnosis comprises an oxonol dye having a polymethine chain. The fluorescent contrast medium for diagnosis is a fluorescent contrast medium for diagnosing the cancer or a fluorescent contrast medium for diagnosing the tumor. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a diagnostic fluorescent contrast agent (hereinafter also referred to as a diagnostic contrast agent or a fluorescent contrast agent) and a fluorescence contrast diagnostic method using the diagnostic fluorescent contrast agent.

  When treating a disease, it is required to detect a morphological change caused in the living body by the disease in an early stage of the disease accurately and quickly by a simple method. Especially when treating cancer, the determination of the location and size of the initial small lesion is essential for early treatment. Examples of methods already known for this purpose include diagnostic imaging, such as endoscopic biopsy, X-ray imaging, MRI and ultrasound imaging. Biopsy is effective in confirming the diagnosis because it can directly observe the lesion, but at the same time, the subject is forced to ache or pain. X-ray imaging and MRI expose the subject to radiation and magnetic fields that are harmful if excessive, and the exposure time increases in proportion to the tracking time when trying to track the time course. Equipment and devices are also large, and a great deal of labor and cost is required for their installation and maintenance. However, near-infrared fluorescence imaging has recently attracted attention as a diagnosis that can be downsized, reduce the labor of operation, and simplify use. It is said that near-infrared light can easily pass through a biological tissue in which 70% or more is made of moisture, and can inspect and diagnose a thickness of 10 cm to 20 cm. Therefore, it has come to be developed as a diagnostic technique for near infrared CT while attracting attention in the field of clinical medicine. In this method, a compound that emits fluorescence when irradiated with excitation light having a near-infrared wavelength of 700 nm to 1000 nm as a contrast agent is irradiated into the living body and has a near-infrared wavelength from outside the body. Irradiation with excitation light, and detection of fluorescence emitted from a compound administered in the body, that is, a so-called fluorescent contrast agent, determines the lesion. As such a fluorescent contrast agent, for example, compounds such as porphyrin compounds and hematoporphyrin that accumulate in tumors are known. These compounds are excited by near-infrared light irradiation to generate triplet oxygen that can oxidize the living body of the lesion, killing cells of the lesion such as cancer, and treating them. Although possible, there is a risk of destroying tissues other than the lesion. On the other hand, contrast methods using known fluorescent dyes such as fluorescein and fluoresamine are known (US Pat. No. 4,945,239), but these fluorescent dyes emit blue to green light with very low light transmission in the living body. This is a problem that can not be detected sufficiently.

Cyanine dyes that emit fluorescence in the near infrared region are expected as fluorescent contrast agents, and various cyanine dye compounds have been studied. Since a fluorescent contrast agent of a cyanine compound was reported, a technique using various peripheral cyanine compounds as a contrast agent has been disclosed in order to change to a compound having a high hydrophilicity, molar extinction coefficient, and quantum yield (for example, Patent Document 1). 2, 3). However, together with the ability to distinguish normal tissue from diseased tissue (contrast power), it is necessary to be completely decomposed and harmless in the living body after contrasting from the living body, or completely discharged (non-accumulating), A safe contrast agent that combines the two has not been found.
JP 2000-95758 A JP-T-2002-526458 JP 2003-160558 A

  An object of the present invention is a cancer that has low water toxicity, excellent water solubility, and emits fluorescence in a region that can be transmitted through living tissue to enable specific imaging of a tumor, cancer, and / or blood vessel. Another object of the present invention is to provide a diagnostic fluorescent contrast agent for tumor and a fluorescent contrast diagnostic method.

  The above object of the present invention is achieved by the following configurations.

  1. A diagnostic fluorescent contrast agent comprising an oxonol dye having a polymethine chain.

  2. 2. The diagnostic fluorescent contrast agent according to 1 above, wherein the diagnostic fluorescent contrast agent is a cancer diagnostic fluorescent contrast agent or a tumor diagnostic fluorescent contrast agent.

  3. 3. The diagnostic fluorescent contrast agent according to 1 or 2 above, wherein the oxonol dye having a polymethine chain is represented by the following general formula (I).

(In the formula, n represents an integer of 3 or 4, Q ₁ and Q ₂ represent a group of atoms forming a carbocyclic or heterocyclic ring, L represents a methine group which may be substituted and may be the same or different. .)
4). 1 to 3 above, wherein the oxonol dye having a polymethine chain represented by the general formula (I) has at least one group selected from a sulfonic acid group, a carboxylic acid group and a phosphoric acid group as an acid group. The diagnostic fluorescent contrast agent according to any one of the above.

  5). The acid group in the oxonol dye having a polymethine chain represented by the general formula (I) has at least one group selected from a sulfonic acid group or a carboxylic acid group, and the total of the acid groups is 2 or more. 5. The diagnostic fluorescent contrast agent according to any one of 1 to 4 above, wherein

  6). 6. The diagnostic fluorescent imaging according to any one of 1 to 5 above, wherein the acid group in the oxonol dye having a polymethine chain represented by the general formula (I) is at least two sulfonic acid groups. Agent.

  7). 7. The diagnostic fluorescent contrast agent according to any one of 1 to 6, wherein the acid group salt is a sodium salt.

  8). The tumor or cancer is at least one in vivo part selected from brain, breast, breast, prostate, colon, lung, liver, pancreas, stomach, lymphoma, uterus, cervix, upper and lower limbs, sarcoma and melanoma 8. The diagnostic fluorescent contrast agent according to any one of 1 to 7, wherein the diagnostic fluorescent contrast agent is provided.

  9. In the fluorescence contrast diagnostic method for diagnosing in vitro using a fluorescent contrast agent, the diagnostic fluorescent contrast agent according to any one of 1 to 8 above is introduced into a living body, and excitation light is irradiated into the living body. A diagnostic method for fluorescence contrast diagnosis, wherein diagnosis is performed in vitro by detecting fluorescence from the diagnostic contrast medium.

  The diagnostic fluorescence contrast agent and the fluorescence contrast diagnosis method according to the present invention have low water toxicity and excellent water solubility, and further emit fluorescence in a region that can be transmitted through living tissue to identify tumors, cancers, and / or blood vessels. It is possible to perform contrasting and has an excellent effect.

  Next, the best mode for carrying out the present invention will be described, but the present invention is not limited thereto.

  Hereinafter, the present invention will be described in detail.

  The oxonol dye having a polymethine chain represented by the general formula (I) preferably used in the present invention will be described.

In the general formula (I), Q ₁ and Q ₂ are carbocycles such as 1,3-cyclopentanedione and 1,3-indandione, and pyrazolone, isoxazolone, barbituric acid, thiobarbituric acid, 3- A group of atoms necessary for forming a heterocyclic ring such as oxythionaphthene, rhodanine, 2-thiohydantoin, 2-thioxazolidinedione, pyrazolopyridine, and quinolone. L represents a methine group, which may be substituted with an alkyl group having 1 to 3 carbon atoms (for example, a methyl group, an ethyl group, etc.), a phenyl group, a benzyl group, a phenethyl group, a hydroxy group, an acyloxy group, a halogen atom, or the like. good. In addition, a substituent of the methine group may form a ring (for example, 5,5-dimethyl-1-cyclohexen-1-yl-3-ylidene).

  The oxonol dye having a polymethine chain represented by the general formula (I) preferably has an acid group (sulfo group, carboxyl group, phosphoric acid group, etc.) in the molecule, and has at least two sulfo groups or carboxy groups. Is more preferable. Examples of the oxonol dye having a polymethine chain represented by the general formula (I) include a pyrazolone oxonol dye represented by the following general formula (I) -1 and barbituric acid and thiobarbi represented by the general formula (I) -2. Examples include tool acid dye compounds. The methine group preferably forms a ring such as 5,5-dimethyl-1-cyclohexen-1-yl-3-ylidene.

In general formula (I) -1. n represents an integer of 3 or 4. R ₁₁ and R ₁₂ represent a hydrogen atom, an alkyl group, an aralkyl group, an aryl group, or a heterocyclic group. Examples of the alkyl group represented by R ₁₁ and R ₁₂ include linear, branched, and cyclic groups such as methyl, ethyl, propyl, isopropyl, n-butyl, and cyclohexyl, and examples of the aralkyl group include benzyl and phenethyl. Examples of the aryl group include phenyl and naphthyl, and examples of the heterocyclic group include benzothiazolyl, pyridyl, pyrimidyl, sulfolanyl, and the like, and an alkyl group, an aralkyl group, and an aryl group are preferable. The alkyl group, aralkyl group, aryl group, and heterocyclic group represented by R ₁₁ and R ₁₂ can have various substituents such as sulfo, carboxy, hydroxy, cyano, halogen (for example, fluorine, chlorine, etc.), alkyl, and the like. (Eg, methyl, isopropyl, trifluoromethyl, t-butyl, ethoxycarbonylmethyl, sulfomethyl, etc.), amino (eg, amino, dimethylamino, sulfoethylamino, piperidino, morpholino, etc.), alkoxy (eg, methoxy, ethoxy, sulfopropoxy, etc.) ), Sulfonyl (eg methanesulfonyl, ethanesulfonyl etc.), sulfamoyl (eg sulfamoyl, dimethylsulfamoyl etc.), acylamino (eg acetamide, benzamide, sulfobenzamide etc.), carbamoyl (Eg, carbamoyl, phenylcarbamoyl, sulfophenylcarbamoyl, etc.), sulfonamide (eg, methanesulfonamide, benzenesulfonamide, etc.), alkoxycarbonyl (eg, ethoxycarbonyl, hydroxyethoxycarbonyl, benzyloxycarbonyl, etc.), aryloxycarbonyl (For example, phenoxycarbonyl, nitrophenoxycarbonyl, etc.), and the like.

The aralkyl group and aryl group represented by R ₁₁ and R ₁₂ preferably have at least one of a sulfo group, a carboxy group and a phosphoric acid group on the aromatic nucleus, and more preferably at least one sulfo group. It is desirable that the group has.

R ₁₃ and R ₁₄ are alkyl groups, aryl groups, aralkyl groups, heterocyclic groups, carboxyl groups, alkoxycarbonyl groups, aryloxycarbonyl groups, carbamoyl groups, ureido groups, thioureido groups, acylamino groups, acyl groups, imide groups, cyano groups. Represents a group, a hydroxy group, an alkoxy group, or an amino group.

Specific examples of the alkyl group, alkoxycarbonyl group, aryloxycarbonyl group, carbamoyl group, acylamino group, alkoxy group, and amino group represented by R ₁₃ and R ₁₄ are the alkyl groups and aralkyls represented by R ₁₁ and R ₁₂ described above. The same thing as the specific example of the substituent introduce | transduced into a group, an aryl group, and a heterocyclic group can be mentioned.

Furthermore, examples of the aryl group represented by R ₁₃ and R ₁₄ include phenyl, sulfopropoxyphenyl, cyanophenyl, carboxyphenyl, nitrophenyl, sulfophenyl, and the like. Aralkyl groups include, for example, benzyl, phenethyl, sulfobenzyl, and the like. For example, furyl, thienyl, etc., ureido groups include, for example, methylureido, phenylureido, etc., thioureido groups, for example, methylthioureido, phenylthioureido, etc., imide groups, for example, succinimide, phthalimide, etc. Examples thereof include acetyl and pivalyl.

As the group represented by R ₁₃ and R ₁₄ , an alkyl group, a carboxy group, an alkoxycarbonyl group, a carbamoyl group, a ureido group, an acylamino group, an acyl group, an imide group, a cyano group, a hydroxy group, an alkoxy group, and an amino group are preferable. Are more preferably an alkyl group, a carboxy group, an alkoxycarbonyl group, a carbamoyl group, an acyl group, a cyano group, an alkoxy group, or an amino group.

In general formula (I) -2, n represents an integer of 3 or 4, W represents an oxygen atom and a sulfur atom, L represents a methine group, R _{21 to} R ₂₄ represent a hydrogen atom, an alkyl group, and an aryl group. Represents an aralkyl group or a heterocyclic group. Examples of the methine group represented by L include those described in the general formula (I).

Examples of the alkyl group represented by R _{21 to} R ₂₄ include the same alkyl groups as R ₁₁ and R ₁₂ listed in the general formula (I) -1, and the alkyl group may have a substituent, As the substituent, for example, the aryl group represented by R ₁₁ and R ₁₂ R _{21 to} R ₂₄ in the general formula (I) -1 is preferably a phenyl group, and the substituent introduced into the phenyl group includes the above-described general formula. (I) -1 includes various substituents introduced into the groups R ₁₁ and R ₁₂ , and at least one of a sulfo group and a carboxy group is present on the aromatic nucleus. It is desirable to have.

The aralkyl group represented by R _{21 to} R ₂₄ is preferably a benzyl group or a phenethyl group, and examples of the substituent introduced onto the aromatic nucleus include the same substituents as the aryl group of R _{21 to} R ₂₄ described above. Can do.

Examples of the heterocyclic group represented by R _{21 to} R ₂₄ include pyridyl, pyrimidyl and the like, and examples of the substituent introduced onto the heterocyclic ring include substitution of the aryl group of R _{21 to} R ₂₄ described above. The same thing as a group can be mentioned.

As the group represented by R _{21 to} R ₂₄ , an alkyl group and an aryl group are preferable, and a barbituric acid and thiobarbituric acid represented by the general formula (I) -2, a carboxyl group in the molecule of the oxonol dye, It is desirable to have at least two of the sulfo groups.

  A method for synthesizing the oxonol dye having a polymethine chain of the present invention will be described.

As a methine chain supplier of an oxonol dye having a polymethine chain represented by the general formula (I) of the present invention,
For example, diamyl compounds as described in P. 249-P251 of “The Cyanine soybeans and related compounds” 1964, Interscience Publishers by Harmer, “The Cyanine, Soybean and Related Compounds” (F. M. Hammer “The cyanines and related compounds” 1964, Interscience Publishers). Can be used.

  The symmetric compound of the present invention can be obtained by reacting 1 mol of methine chain donor with 2 mol of an acidic mother nucleus compound such as pyrazolone. For an asymmetric type compound, first, 1 mol of a methine chain donor and 1 mol of a mother nucleus compound are reacted to synthesize an aminopolymethine-substituted mother nucleus compound, and then another 1 mol of an appropriate mother nucleus compound is reacted. Can be synthesized. These condensation reactions are described in detail, for example, in JP-B-54-38129, JP-B-52-38056, JP-A-49-99620, JP-A-50-91627, etc., and can be referred to.

  Specific examples of the oxonol dye having a polymethine chain represented by the general formula (I) are shown below, but the specific examples of the present invention are not limited thereto.

  It is important that the fluorescent contrast agent to be used in the living body is not accumulated in the body but is quickly discharged out of the body, and is basically a water-soluble condition. The means for improving water solubility is preferably anionic carboxylic acid or sulfonic acid salt, and the near-infrared fluorescent contrast agent of the present invention introduces three or more sulfonic acid groups into the above compound. Thus, the water solubility is remarkably improved.

  In order to obtain excellent water solubility, the number of sulfonic acid groups is preferably 4 or more. In order to facilitate the synthesis, the number of sulfonic acid groups is 10 or less, preferably 8 or less. A measure of water solubility can be determined by measuring the partition coefficient of each compound, for example, by measuring the partition coefficient in a two-phase system of an aliphatic alcohol such as butanol and water. For example, by introducing 3 or more sulfonic acid groups, the distribution coefficient logPo / w of n-butanol / water becomes −1.00 or less.

  A method for determining water solubility in a living body is that it dissolves in physiological saline and does not precipitate or precipitate after a lapse of time at 36 ° C. Any salt that is acceptable when administered in vivo may be any salt that forms a non-toxic salt with the oxonol dye having the polymethine chain of general formula (I).

  Examples thereof include alkali metal salts such as sodium salts and potassium salts; alkaline earth metal salts such as magnesium salts and calcium salts, tryptophan, methionine, lysine, phenylalanine, leucine, isoleucine, valine, threonine and arginine. And amino acid salts such as salts thereof. Particularly preferred are sodium salts that have low toxicity in vivo.

  The diagnostic contrast medium of the present invention is injected into a living body by means such as injection, spraying or application into blood vessels (veins, arteries), oral, intraperitoneal, subcutaneous, intradermal, intravesical, intratracheal (branch), etc. Can be administered.

  Specifically, the diagnostic fluorescent contrast agent used in the extracorporeal fluorescence contrast diagnostic method of the present invention comprises an oxonol dye having a polymethine chain represented by the general formula (I) of the present invention, distilled water for injection, and physiological saline. It is suspended or dissolved in water or a solvent such as Ringer's solution, and pharmacologically acceptable additives such as carriers and excipients may be added as necessary. These additives include substances such as pharmacologically acceptable electrolytes, buffers, detergents, and substances for adjusting osmotic pressure to improve stability and solubility (eg, cyclodextrins, liposomes, etc.). . Various additives commonly used in related fields can be used. The fluorescent contrast medium according to the present invention is preferably manufactured through sterilization treatment for medical use. The fluorescent contrast agent can be administered to the living body site by injection, injection or application, intravascular (vein, artery), oral, intraperitoneal, transdermal, subcutaneous, intravesical or intrabronchial administration. Preferably, the contrast agent is administered into the blood vessel in the form of a solution, emulsion or suspension.

  The dosage of the diagnostic contrast medium of the present invention is not particularly limited as long as it is an amount that can finally detect the site to be diagnosed. The type of the compound emitting near-infrared fluorescence to be used, the age and body of the subject to be administered The dose can be appropriately increased or decreased depending on the size of the organ and the target organ, but is usually 0.1 to 100 mg / kg · body weight, preferably 0.5 to 20 mg / kg · body weight.

  The diagnostic contrast agent of the present invention can also be suitably used as an animal contrast agent, and its administration form, administration route, dosage and the like are appropriately selected according to the body weight and state of the subject animal.

  In the present invention, a series of compounds having two or more, preferably four or more sulfonic acid groups in the oxonol dye molecule having a polymethine chain represented by the general formula (I) can be applied to tumor tissue at a certain concentration. It accumulates and has the property of being easily discharged outside the body when the concentration falls below a certain concentration, and it can be used as a fluorescent contrast agent capable of selectively and specifically imaging a tumor tissue using the characteristics.

  Further, the compound of the present invention is difficult to diffuse out of the blood vessel wall once injected into the blood vessel, and has a high property of remaining in the blood vessel, and can also be used as an angiographic agent.

  The fluorescent contrast method of the present invention is characterized by using the fluorescent contrast agent of the present invention. The measurement method is performed using a method known to those skilled in the art. The conditions of the excitation wavelength, the fluorescence wavelength for detection, and the like are optimized so as to obtain the optimal and best detection state. It is determined appropriately according to the type, subject to be administered and the like.

  The time required to start the measurement using the fluorescence contrast method of the present invention after administering the fluorescent contrast agent of the present invention to the measurement object also varies depending on the type of the fluorescent contrast agent to be administered, the subject to be administered, etc. For example, when administered for the purpose of tumor or cancer imaging, it is preferable to select an elapsed time of 10 minutes to 6 hours after administration. If the elapsed time is less than 10 minutes, the fluorescence is unevenly distributed on the whole and it is difficult to distinguish between the target site and the other site, and if it exceeds 6 hours, the contrast medium is excreted outside the body. For the purpose of angiography, it is preferable to measure at an elapsed time of 1 hour after administration.

  The extracorporeal fluorescent contrast diagnostic method of the present invention is characterized by the use of the fluorescent contrast agent of the present invention. This method is performed according to a known method, and parameters such as the excitation wavelength and the fluorescence wavelength to be detected are appropriately determined so as to be optimal depending on the type of the near-infrared fluorescent contrast agent to be administered and the administration target. The The time taken from the administration of the fluorescent contrast agent of the present invention to the measurement object to the start of measurement by the fluorescence imaging method of the present invention varies depending on the type of near-infrared fluorescent contrast agent used and the administration object. For example, for tumor imaging purposes, the disappearance time is considered to be approximately 4-120 hours after administration. If the disappearance time is too short, the fluorescence is too strong to clearly separate the target site from other sites. If the length is too long, the contrast medium may be excreted from the body. When angiography is required, detection is performed immediately after administration or within about 30 minutes.

  In this method, the fluorescent contrast agent is administered to a detection target, and the detection target is exposed to excitation light from an excitation light source. Next, the fluorescence generated by the excitation light from the near-infrared fluorescent contrast agent is detected by a fluorescence detector. The excitation wavelength varies depending on the near-infrared fluorescent contrast agent used, but is not limited as long as the compound emits fluorescence effectively in the near-infrared region. Preferably, near-infrared light having excellent biological permeability is used.

  The wavelength for exposure is preferably 200 to 1010 nm, more preferably 600 to 1000 nm, and still more preferably 700 to 850 nm, and fluorescence is detected with high sensitivity. In this case, as the fluorescence excitation light source, various laser light sources such as an ion laser, a dye laser, a semiconductor laser, or a normal excitation light source such as a halogen light source or a xenon light source may be used. Various optical filters can be used to obtain. Similarly, when detecting fluorescence, the fluorescence detection sensitivity can be increased by using various optical filters that select only fluorescence from the fluorescent contrast agent.

  The detected fluorescence is data-processed as fluorescence information and used to generate a recordable fluorescence image. The fluorescence image is generated by irradiating a wide area including the target tissue, detecting fluorescence with a CCD camera, and processing the obtained fluorescence information. Alternatively, an optical CT apparatus, an endoscope, or a fundus camera may be used. The fluorescence imaging method of the present invention can visualize systemic diseases, tumors, blood vessels and the like without damaging the living body.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated concretely, the embodiment of this invention is not limited to these.

Example 1
Production of Breast Cancer Carcinoma Model Mice Breast Cancer Carcinogenesis Model Mice Production Senescence Accelerated Mouse, a Carcinogenic Substance 7,12-Dimethylbenz [a] anthracene (DMBA) ) Was administered to produce a breast cancer model mouse. The method for carcinogenesis in mice was performed according to Japanese Patent Application Laid-Open No. 2003-033125. 20 SAMP6 / Ta mice were administered to DMBA at a dose of 0.5 mg / mouse / week for a total of 6 times. As feed, high protein and high calorie CA-1 solid (manufactured by Claire Japan) was given. After the sixth administration of the carcinogen, the period from the first administration up to the 20th week was defined as a drug holiday. Breast cancer and lung metastases of breast cancer were searched histopathologically. DMBA was administered 6 times at 1-week intervals, then the drug was withdrawn from the start of the subsequent administration until the 20th week, and mice with breast cancer (75% breast cancer incidence) were used.

Fluorescence Contrast Test Tumor tissue fragment (2 mm × 2 mm square) of mouse breast cancer was transplanted subcutaneously into the breast of the left breast of BALB / c nude mice (5 weeks old, Claire Japan). Ten days later, when the tumor grew to a diameter of about 5 mm, the mouse was subjected to the test. A titanium sapphire laser was used as the fluorescence excitation light source. Using a ring-type light guide (Sumita Optical Glass Co., Ltd.), the test mice were uniformly irradiated with laser light so that the dispersion of irradiation was within 2%. The irradiation output was adjusted to be about 36 μW / cm ² near the skin surface of the mouse. Fluorescence was excited at the maximum excitation wavelength of each compound, and fluorescence emission from the mouse was detected and imaged through a short wavelength cut filter using a CCD camera (C4880, Hamamatsu Photonics). The cut filter was selected to match the excitation wavelength of the compound (800 nm to 900 nm). The irradiation time was adjusted according to the fluorescence intensity of each compound.

  As a fluorescent contrast agent, each test compound was dissolved in distilled water (0.5 mg / ml) and administered to the mice from the milk ducts and the filaments. The dose was 1 mg / kg each. Twelve minutes after compound administration, the mice were anesthetized with diethyl ether, and a fluorescent image of the whole mouse was imaged. As a sensor, a photodiode (wavelength range: 220 to 1010 nm) of a fluorometer (JASCO FP-6600) was used.

  The conditions are shown below, and the fluorescence sensitivity is expressed as a relative sensitivity with a comparison of 100. It was displayed as a logarithm of the amount of fluorescence generated relative to the reference. The amount of the contrast medium discharged from the body is defined as 100 after the contrast medium is administered, and the accumulated amount discharged from the urethra by the catheter 13 weeks later is added to the body and discharged from the liquid chromatograph 2010A (Shimadzu Corporation). The amount was calculated and the emission rate relative to the initial concentration was determined. The results are shown in Table 1. In addition, the compound shown by No. 50 of the special table 2002-526458 was used as a contrast medium for comparison. In the water solubility test of the compound, 0.5 mg was dissolved in 1 ml of 0.9% physiological saline and left standing at 36 ° C. for 2 weeks to confirm precipitation and precipitation.

◎: No level at all
○: Level that appears to be slightly haze, but disappears by stirring,
Δ: Haze is applied, but the level is not lost by stirring.
×: Level of precipitation,
As evaluated.

  The diagnostic fluorescent contrast agent containing an oxonol dye having a polymethine chain represented by the general formula (I) of the present invention is superior in water solubility, fluorescence sensitivity, and extracorporeal excretion rate compared to a comparative fluorescent contrast agent, It turns out to be effective for the diagnosis of tumors or cancers.

Claims (9)

A diagnostic fluorescent contrast agent comprising an oxonol dye having a polymethine chain. The diagnostic fluorescent contrast agent according to claim 1, wherein the diagnostic fluorescent contrast agent is a fluorescent contrast agent for cancer diagnosis or a fluorescent contrast agent for tumor diagnosis. The diagnostic contrast agent according to claim 1 or 2, wherein the oxonol dye having a polymethine chain is represented by the following general formula (I).

(In the formula, n represents an integer of 3 or 4, Q ₁ and Q ₂ represent a group of atoms forming a carbocyclic or heterocyclic ring, L represents a methine group which may be substituted and may be the same or different. .) The oxonol dye having a polymethine chain represented by the general formula (I) has at least one group selected from a sulfonic acid group, a carboxylic acid group and a phosphoric acid group as an acid group. The diagnostic fluorescent contrast agent according to any one of the above. The acid group in the oxonol dye having a polymethine chain represented by the general formula (I) has at least one group selected from a sulfonic acid group or a carboxylic acid group, and the total of the acid groups is 2 or more. The diagnostic fluorescent contrast agent according to any one of claims 1 to 4, wherein The diagnostic fluorescence according to any one of claims 1 to 5, wherein the acid group in the oxonol dye having a polymethine chain represented by the general formula (I) is at least two sulfonic acid groups. Contrast agent. The diagnostic fluorescent contrast medium according to any one of claims 1 to 6, wherein the salt of the acid group is a sodium salt. The tumor or cancer is at least one in vivo part selected from brain, breast, breast, prostate, colon, lung, liver, pancreas, stomach, lymphoma, uterus, cervix, upper and lower limbs, sarcoma and melanoma The diagnostic fluorescent contrast agent according to any one of claims 1 to 7, wherein the diagnostic fluorescent contrast agent is provided. In a fluorescence contrast diagnostic method for diagnosing in vitro using a fluorescent contrast agent, the diagnostic fluorescent contrast agent according to any one of claims 1 to 8 is introduced into a living body, and excitation light is irradiated into the living body. And diagnosing in vitro by detecting fluorescence from the diagnostic contrast agent.