JPWO2020066577A1 - Periodontal disease remedy - Google Patents

Abstract

（式中、Ｒ^１は、水素原子又はアシル基を表し、Ｒ^２は、水素原子、アルキル基、シクロアルキル基、アリール基又はアラルキル基を表す。）で表される化合物又は薬学的に許容されるその塩を含む歯周病の治療薬を、歯周病患部に投与し、その後３６０ｎｍ〜７００ｎｍの波長の光を照射する。An object of the present invention is to provide a therapeutic agent and a therapeutic method that enable PDT even for periodontal disease pathogens that are not effective by conventional photodynamic therapy (PDT).
The following formula (I)
[Chemical 1]

(In the formula, R ¹ represents a hydrogen atom or an acyl group, and R ² represents a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group or an aralkyl group) or a pharmaceutically acceptable compound. A therapeutic agent for periodontal disease containing a salt thereof is administered to the affected part of the periodontal disease, and then light having a wavelength of 360 nm to 700 nm is irradiated.

Description

The present invention contains 5-aminolevulinic acid (hereinafter sometimes referred to as "ALA") or a derivative thereof or a salt thereof, and irradiates light having a wavelength of 360 nm to 700 nm ALA-photodynamic therapy (hereinafter referred to as "ALA"). -PDT "), which relates to a therapeutic agent for periodontal disease.

In recent years, photodynamic therapy (hereinafter referred to as PDT), which is a method of treating a target site by administering a compound that reacts with light and irradiating with light, has been developed. PDT is attracting attention as a new cancer treatment method considering Quality Of Life (QOL) because it is easy to treat, has low bioinvasiveness, and can preserve organs. ..

ALA, one of the drugs used for PDT, is known as an intermediate in the pigment biosynthesis pathway widely present in animals, plants and fungi, and is usually biosynthesized from succinyl-CoA and glycine by ALA synthesizer. .. Although ALA itself is not photosensitive, it is metabolically activated into protoporphyrin IX (hereinafter also referred to as "PpIX") by a series of enzymes in the heme biosynthetic pathway in cells, and is directly directed to tumor tissue and new blood vessels. It is known that when the PpIX accumulation site is accumulated and the PpIX accumulation site is irradiated with laser light, cancer cells are denatured and necrotic by active oxygen species typified by singlet oxygen and / or hydroxyl radical, superoxide, etc. generated by excitation of light. Has been done.

Since Professor Kennedy of Queens University of Canada reported in 1986 that skin cancer can be treated by applying ALA and irradiating it with light (see, for example, Non-Patent Document 1), various sites using ALA have been used. Diagnosis and treatment methods for lesions of ALA have been reported. For example, it was developed based on the finding that when ALA is administered into the body, PpIX induced from ALA accumulates in cancer and emits fluorescence by light irradiation. Tumor diagnostic agents and the like have been proposed (see, for example, Patent Documents 1 and 2).

On the other hand, in the field of periodontal disease treatment, as phototherapy in a broad sense, treatment using various light energies such as high-power laser has been performed for about 20 years, and recently, light Antimicrobial photodynamic therapy (a-PDT), which uses a photochemical reaction in combination with a dye, is attracting attention as a new means.

Enterococcus faecalis (E. faecalis), which is one of the causative bacteria of periodontal disease, was cultured in sterilized single root teeth using the culture solution AC Broth (manufactured by Aldrich) for 4 weeks, and in a 100 μmol methylene blue solution. After washing, it was irradiated with light of 660 nm. It is known that Fecalis does not die (see Patent Document 3).

In addition, semiconductor lasers with a higher energy wavelength of 405 nm were used by Porphyromonas gingivalis (P. gingivalis), Prevotella intermedia (P. intermedia), which are the causative agents of periodontal disease. When Fecalis was irradiated, P.I. Gingivalis and P.I. In the intermedia, the number of cells decreased, but E. It is known that the number of cells of Fecalis did not decrease (Non-Patent Document 2).

Japanese Patent No. 2731032 Japanese Unexamined Patent Publication No. 2006-124372 Special Table 2017-534412

J.C Kennedy, R.H Pottier and DC Pross, Photodynamic therapy with endogeneous protoprophyrin IX: basic principles and present clinical experience, J. Photochem., Photobiol. B: Biol., 6 (1990) 143-148 International Journal of Photoenergy, Volume 2014, Article ID 387215, 7 pages

PDT is non-invasive, has almost no side effects or physical pain to the patient, and although it is a safe and easy treatment method, some of the bacteria that cause periodontal disease are not effective by conventional methods. I faced the problem of being there.
An object of the present invention is to provide a therapeutic agent, a therapeutic method, etc. that enable PDT even for periodontal disease pathogens that are not effective with conventional PDT.

The present inventors have described E.I. It is thought that the reason why Fecalis does not die even when irradiated with light is that PpIX cannot be biosynthesized in the cells, and E.I. Fecalis was cultivated and irradiated with light, but the bacteria did not die. Therefore, as a result of various studies on the culture conditions, E.I. It was found that Fecalis could be killed, and the present invention was completed.

（式中、Ｒ^１は、水素原子又はアシル基を表し、Ｒ^２は、水素原子、アルキル基、シクロアルキル基、アリール基又はアラルキル基を表す。）で表される化合物又は薬学的に許容されるそれらの塩（以下「ＡＬＡ類」ということがある。）を含み、歯周病患部に式（I）で表される化合物又は薬学的に許容されるそれらの塩を投与し、その後３６０ｎｍ〜７００ｎｍの波長の光を照射する５−アミノレブリン酸−光線力学的療法のための歯周病の治療薬。
（２）歯周病における歯周病原因菌が、Ｅ．フェカリスである（１）に記載の治療薬。
（３）３６０ｎｍ〜７００ｎｍの波長の光が、青色発光ダイオードである（１）に記載の治療薬。
また本発明の他の実施態様は以下のとおりである。
（４）上記式（I）で表される化合物又は薬学的に許容されるそれらの塩を歯周病患部に投与し、その後３６０ｎｍ〜７００ｎｍの波長の光を照射する歯周病の治療方法。
（５）歯周病患部に投与し、その後３６０ｎｍ〜７００ｎｍの波長の光を照射する歯周病の治療に用いるための上記式（I）で表される化合物又は薬学的に許容されるそれらの塩。
（６）歯周病患部に投与し、その後３６０ｎｍ〜７００ｎｍの波長の光を照射する歯周病の治療薬の製造に用いるための上記式（I）で表される化合物又は薬学的に許容されるそれらの塩の利用。That is, the present invention is as follows specified by the following matters.
(1) The following formula (I)

(In the formula, R ¹ represents a hydrogen atom or an acyl group, and R ² represents a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group or an aralkyl group) or a pharmaceutically acceptable compound. These salts (hereinafter sometimes referred to as "ALAs") are contained, and the compound represented by the formula (I) or pharmaceutically acceptable salts thereof is administered to the affected part of the periodontal disease, and then 360 nm. 5-Aminolevulinic acid-a therapeutic agent for periodontal disease for photodynamic therapy, which irradiates light with a wavelength of ~ 700 nm.
(2) Periodontal disease-causing bacteria in periodontal disease are E.I. The therapeutic agent according to (1), which is Fecalis.
(3) The therapeutic agent according to (1), wherein the light having a wavelength of 360 nm to 700 nm is a blue light emitting diode.
In addition, other embodiments of the present invention are as follows.
(4) A method for treating periodontal disease in which a compound represented by the above formula (I) or a pharmaceutically acceptable salt thereof is administered to the affected area of periodontal disease and then irradiated with light having a wavelength of 360 nm to 700 nm. ..
(5) Compounds represented by the above formula (I) or pharmaceutically acceptable compounds for use in the treatment of periodontal disease, which are administered to the affected area of periodontal disease and then irradiated with light having a wavelength of 360 nm to 700 nm. Salt.
(6) The compound represented by the above formula (I) or pharmaceutically acceptable for use in the production of a therapeutic agent for periodontal disease, which is administered to the affected area of periodontal disease and then irradiated with light having a wavelength of 360 nm to 700 nm. The use of those salts to be done.

Treatment of periodontal disease using the therapeutic agent for periodontal disease of the present invention is non-invasive, has almost no side effects or physical pain to the patient, is safe and easy, and is ineffective by conventional methods. Periodontal disease caused by bacteria that cause periodontal disease can also be treated.

FIG. 1 shows E.I. After culturing Fecalis under anaerobic conditions by adding ALA to various concentrations, the measurement results by the dilution plate method for measuring the number of cells in the culture solution obtained by irradiating with light are shown. show. FIG. 2 shows E.I. After culturing Fecalis under aerobic conditions by adding ALA and ALA + EDTA to various concentrations, a dilution plate method is used to measure the number of cells in the culture solution obtained by irradiating with light. The measurement result is shown.

The therapeutic agent for periodontal disease of the present invention is not particularly limited as long as it is a therapeutic agent for ALA-PDT that contains ALA and is used in ALA-PDT that irradiates light having a wavelength of 360 nm to 700 nm. 5-Aminolevulinic acid-photodynamic diagnosis (ALA-PDD) to detect PpIX accumulation sites by irradiating excitation light with a wavelength of 360 nm to 420 nm before ALA-PDT irradiating light with a wavelength of 360 nm to 700 nm. However, a therapeutic agent that does not require such ALA-PDD can be particularly preferably exemplified. Further, the treatment system to which the therapeutic agent for periodontal disease of the present invention is applied may be any system provided with an ALA-PDT device, and may be provided with an ALA-class administration device or an ALA-PDD. ..

In the compound represented by the formula (I), R ¹ represents a hydrogen atom or an acyl group, and R ² represents a hydrogen atom, a linear or branched alkyl group, a cycloalkyl group, an aryl group or an aralkyl group.

Examples of the acyl group in R ¹ include linear or branched carbons such as formyl group, acetyl group, propionyl group, butyryl group, isobutyryl group, valeryl group, isovaleryl group, pivaloyl group, hexanoyl group, octanoyl group and benzylcarbonyl group. Examples thereof include an alkanoyl group having a number of 1 to 8 and an aloyl group having 7 to 14 carbon atoms such as a benzoyl group, a 1-naphthoyl group and a 2-naphthoyl group. In the present invention, the acyl group includes an alkoxycarbonyl group such as a methoxycarbonyl group, an ethoxycarbonyl group, an n-propoxycarbonyl group, or an isopropoxycarbonyl group for convenience.

The alkyl group in R ^2, a methyl group, an ethyl group, a propyl group, an isopropyl group, butyl group, isobutyl group, sec- butyl group, tert- butyl group, a pentyl group, an isopentyl group, a neopentyl group, a hexyl group, a heptyl group , A linear or branched alkyl group having 1 to 8 carbon atoms such as an octyl group can be mentioned.

As ^{the cycloalkyl group in R 2,} there are saturated or partially unsaturated bonds such as a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, a cyclooctyl group, a cyclododecyl group, and a 1-cyclohexenyl group. Cycloalkyl groups having 3 to 8 carbon atoms can be mentioned.

Examples ^{of the aryl group in R 2} include an aryl group having 6 to 14 carbon atoms such as a phenyl group, a naphthyl group, an anthryl group and a phenanthryl group.

The aralkyl group in R ^2, the aryl moiety can be the same example as above aryl group, the alkyl moiety can be the same example as above alkyl group, specifically, a benzyl group, phenethyl group, phenylpropyl group, phenylbutyl group , Benzhydryl group, trityl group, naphthylmethyl group, naphthylethyl group and other aralkyl groups having 7 to 20 carbon atoms can be mentioned.

The above R ¹ and R ² may have a substituent within a chemically acceptable range, if necessary, and examples of such a substituent include a halogen atom, an alkyl group, a haloalkyl group, and an alkoxy group. , Nitro group, aryl group and the like.

As the compound represented by the formula (I), it is preferable to act as any ALA derivative of ALA capable of forming PpIX in vivo, for example, an ALA prodrug or an intermediate capable of forming ALA in vivo. Examples of the ALA prodrug which is converted (for example, enzymatically) to porphyrin without forming ALA as a body are mentioned, and among them, ALA ester is preferably mentioned.

Examples of ALA esters include ALA methyl ester, ALA ethyl ester, ALA n-propyl ester, ALA n-butyl ester, ALA n-pentyl ester, ALA n-hexyl ester, ALA n-octyl ester, and ALA 2-methoxyethyl ester. , ALA 2-methyl-n-pentyl ester, ALA 4-methyl-n-pentyl ester, ALA 1-ethyl-n-butyl ester, ALA 3,3-dimethyl-n-butyl ester, ALA benzyl ester, ALA 4- Isopropylbenzyl ester, ALA 4-methylbenzyl ester, ALA 2-methylbenzyl ester, ALA 3-methylbenzyl ester, ALA 4- (t-butyl) benzyl ester, ALA 4- (trifluoromethyl) benzyl ester, ALA 4- Methoxybenzyl ester, ALA 3,4- (dichloro) benzyl ester, ALA 4-chlorobenzyl ester, ALA 4-fluorobenzyl ester, ALA 2-fluorobenzyl ester, ALA 3-fluorobenzyl ester, ALA 2,3,4 5,6-Pentafluorobenzyl ester, ALA 3-nitrobenzyl ester, ALA 4-nitrobenzyl ester, ALA 2-phenylethyl ester, ALA 4-phenylbutyl ester, ALA 3-pyridyl-methyl ester, ALA 4-phenylbenzyl Esters, N-[(1-acetyloxy) ethoxycarbonyl] ALA benzyl ester, N-formyl-ALA methyl ester, N-acetyl-ALA ethyl ester, N-propionyl-ALA methyl ester, N-butyryl-ALA methyl ester, Examples thereof include N-formyl-ALA ethyl ester, N-acetyl-ALA ethyl ester, N-propionyl-ALA ethyl ester, and N-butyryl-ALA ethyl ester.

The compound represented by the formula (I) can be administered as various salts for increasing solubility depending on the form of administration. Examples of the salt of the compound represented by the formula (I) include a pharmacologically acceptable acid addition salt, metal salt, ammonium salt, organic amine addition salt and the like. As acid addition salts, for example, each inorganic acid salt such as hydrochloride, hydrobromide, hydroiodide, phosphate, nitrate, sulfate, formate, acetate, propionate, toluenesulfonic acid Salt, succinate, oxalate, lactate, tartrate, glycolate, methanesulfonate, butyrate, valerate, citrate, fumarate, maleate, malate, etc. Examples thereof include organic acid addition salts. Examples of the metal salt include alkali metal salts such as lithium salt, sodium salt and potassium salt, alkaline earth metal salts such as magnesium and calcium salt, and metal salts such as aluminum and zinc. Examples of the ammonium salt include alkylammonium salts such as ammonium salt and tetramethylammonium salt. Examples of the organic amine salt include each salt such as triethylamine salt, piperidine salt, morpholine salt, and toluidine salt. These salts can also be used as a solution at the time of use.

As the ALA derivative, various esters such as ALA or ALA methyl ester, ALA ethyl ester, ALA propyl ester, ALA butyl ester and ALA pentyl ester, and these hydrochlorides, phosphates and sulfates are preferably mentioned, and ALA is preferable. Hydrochlorides and ALA phosphates are particularly preferred.

The ALA derivative can be produced by any known method of chemical synthesis, microbial production, or enzymatic production. In addition, the ALA derivative may form a hydrate or a solvate, and either of them may be used alone or in combination of two or more.

When the above ALA derivative is prepared as an aqueous solution, care must be taken not to make the aqueous solution alkaline in order to prevent decomposition of the ALA derivative. If it becomes alkaline, it can be prevented from decomposing by removing oxygen.

In addition to the ALAs, the following photosensitizers can be mixed and used or co-administered as needed.
Hematoporphyrin derivatives (HpD); Hematoporphyrins such as Photofrin (registered trademark) (Quadra Logic Technologies, Vancouver, Canada), Hematoporphyrin IX (HpIX); Siehoff, Wesselbrenerkov, Germany); Tetra (m-hydroxyphenyl) porphyrin (m-THPC), Bacterioporphyrin (Scotia Pharmaceutical Company, Sally, UK), Mono-L-Asparatyl Porphyrin e6 (NPe6) (Japan) Petrochemical Company, California, USA), Chlorin e6 (Porphyrin Products), Benzoporphyrin (Quadra Logic Technologies, Vancouver, Canada) (eg, benzoporphyrin derivative monoacid ring A, BPD-MA), Purpurine (PDT) Porphyrins such as pharmaceutical companies, California, USA (eg tin-ethyl etiopurpurin, SnET2); phthalocyanine (eg zinc- (Quadralogic Technologies, Vancouver, Canada), some aluminum -Or silicon phthalocyanine, which may be sulfonated, in particular _{sulfonated phthalocyanine such as aluminum phthalocyanine disulfonic acid (AlPcS 2a} ), aluminum phthalocyanine tetrasulfonic acid (AlPcS ₄ ); porphysen; hypocrellin; protoporphyrin. IX (PpIX); hematoporphyrin diester; uroporphyrin; coproporphyrin; juuteroporphyrin; polyhematoporphyrin (PHP) and its precursors and derivatives; tetracycline (eg, Topicycline®), Shire (Shire)).
These can be used alone or in combination of two or more.

In addition, ALAs can be administered with compounds having other activities that can enhance the photosensitizing effect and thus promote PDT. Examples of such compounds include chelating agents, and more specifically, in the literature on aminopolycarboxylic acid, metal detoxification, or the literature on chelation of paramagnetic metal ions in contrasting agents used in magnetic resonance imaging. Examples thereof include the listed chelating agents, and more specifically, ethylenediamine-N, N, N', N'-tetraacetic acid (EDTA), 1,2-diaminocyclohexane-N, N, N', N. '-Tetraacetic acid (CDTA), diethylenetriamine-N, N, N', N'', N'-pentetic acid (DTPA), 1,4,7,10-tetraazacyclododecane-1,4,7, Examples thereof include 10-tetraacetic acid (DOTA), desferrioxamine or well-known derivatives / analogs thereof, which can be used alone or in admixture of two or more.
When having a chelating agent, the chelating agent is typically used at a concentration of 0.05 to 20% (w / v), for example at a concentration of 0.1 to 10% (w / v).

In the present invention, administration of ALAs to a periodontal disease-affected portion means local administration to or around the periodontal disease-affected portion, and topical application is particularly preferable. Topical administration into the periodontal pocket can be performed by using techniques known in the art, such as syringes, catheters or other suitable drug delivery systems.

Examples of the dosage form of the therapeutic agent of the present invention include gels, creams, ointments, sprays, lotions, aerosols, and external liquids.

In the therapeutic agent of the present invention, the concentration of ALAs contained varies depending on various factors including the chemical properties, chemical composition, administration method and degree of the disease to be treated, but is 20% (w). A concentration range of less than / v) is preferred, more preferably 0.05 to 16% (w / v), particularly preferably 0.5 to 14% (w / v), for example 1.5 to 12.0%. A range of (w / v) or 2-10% (w / v) can be preferably exemplified.

The method for treating periodontal disease of the present invention is itself photosensitized when a light-responsive compound is administered to a periodontal disease-causing bacterium and PDT for treating periodontal disease is performed by irradiating with light. ALAs that have no action were administered to the affected area of periodontal disease, and PpIX induced via the pigment biosynthesis pathway accumulated in the cells of the periodontal disease-causing bacteria and accumulated in the periodontal disease-causing bacteria cells. By irradiating PpIX with light to excite it, surrounding oxygen molecules are photoexcited, and the active oxygen species typified by single term oxygen and / or hydroxyl radical, superoxide, etc. generated by the excitation of the resulting light , It is a treatment method for periodontal disease utilizing the fact that it has a cell-killing effect due to its strong oxidizing power.

The therapeutic agent of the present invention is administered in and around the affected area of periodontal disease, particularly in the periodontal pocket, and a specific time elapses before the site to be treated for obtaining the desired photosensitizing effect is exposed. It is preferable to do so. Prior to exposure, excess therapeutic agent is preferably removed.

The time from administration to exposure depends on the type of ALA, the conditions to be treated and the form of administration. The time is, for example, about 3 to 6 hours, preferably 0 to 90 minutes, 5 to 90 minutes, 30 to 90 minutes, and particularly preferably 10 to 50 minutes.

After the therapeutic agent of the present invention is administered to a periodontal disease-affected portion such as a periodontal pocket, the periodontal disease-affected portion such as in the periodontal pocket needs to be placed under anaerobic conditions. In the present invention, to put the periodontal disease-affected part under anaerobic conditions means to put the periodontal disease-affected part under anaerobic conditions when the periodontal disease-affected part is not under anaerobic conditions. If the periodontal diseased area is already under anaerobic conditions, maintain that state, and even if the periodontal diseased area is already under anaerobic conditions, the present invention When the anaerobic condition disappears when the therapeutic drug is administered, it means to return to the original state or to bring the anaerobic condition. In order to keep the periodontal disease-affected part such as the periodontal pocket under anaerobic conditions, for example, a method of covering the area around the periodontal disease-affected part with a material capable of blocking oxygen can be considered. Further, since the periodontal disease-affected portion is originally a portion close to the tooth root and the environment in which the periodontal disease-causing bacteria are propagated is anaerobic, the present invention includes the portion in which the periodontal disease-causing bacteria are propagated. It is also conceivable to use physical or chemical methods to allow the therapeutic agent to penetrate.

After administration of the therapeutic agent of the present invention, photoactivation can be performed using a light source known in the art, for example, a light emitting diode such as a blue LED or a red LED, a semiconductor such as a blue semiconductor laser or a red semiconductor laser. A laser, a discharge lamp having a strong blue or red emission spectrum, and the like can be mentioned, but a blue LED or a red LED can be preferably exemplified because the device becomes compact and is advantageous in terms of cost and portability. can. The wavelength of the light used for irradiation can be selected in order to obtain a more efficient photosensitization effect, and examples thereof include light in the range of 360 to 700 nm. The energy density is preferably in the range of 10~200J / cm ^2, more preferably in the range of 10 to 100J / cm ^2, the range of 20~60J / cm ² is particularly preferred.

The power density of the light source used, if the range the effects of the present invention is not particularly restricted, for example, preferably in the range of ^{15~400mW / cm 2, 15~50mW / cm} 2, 5~40mW / cm ^{The range of 2} , 10 to 35 mW / cm ² is more preferable, and the range of 15 to 35 mW / cm ² is particularly preferable.

Further, the irradiation light may be continuous light or pulsed light, but pulsed light is more preferable in that damage to a normal skin surface can be reduced by using pulsed light.

The light irradiation time depends on the energy density and the power density, but is preferably 5 to 30 minutes, preferably 15 minutes. The irradiation may be performed only once, or for example, the irradiation interval may be 1 to 10 minutes, and the light irradiation amount may be divided into several parts and used as light irradiation.

Examples of the excitation light irradiation device include a small-diameter optical fiber for a light source. A semiconductor laser or LED light source having a strong radiation illuminance to enable excitation of PpIX and a narrow irradiation area to enable accurate automatic identification is preferable, and the excitation light is guided to the outside from one end. It is preferable to have an excitation light light source unit that emits light to the light source, and specific examples of the excitation light light source unit include a small-diameter optical fiber.

As described above, in the treatment method using the therapeutic agent of the present invention, ALA-PDD can also be performed. ALA-PDD uses the fact that PpIX accumulated in the cells of the periodontal disease-causing bacteria emits red fluorescence when the PpIX accumulated in the cells of the periodontal disease-causing bacteria is irradiated with purple light before the ALA-PDT of the present invention. This is a determination method for identifying the attachment site. Examples of the ALA-PDD device used in the ALA-PDD step include an excitation light irradiation device of PpIX, a red fluorescence detection device peculiar to PpIX in an excited state, or a device in which these are integrated. As the light emitted from the PpIX excitation light irradiation device, light having a wavelength at which red fluorescence peculiar to PpIX can be observed by exciting PpIX is preferable, and purple close to ultraviolet light belonging to the absorption peak of PpIX belonging to the so-called Soret band. The light having a wavelength in the range of at least 360 nm to 420 nm may be used, and examples thereof include 360 to 420 nm and 403 to 410 nm, with 408 nm being preferable.

As the light source for irradiating the excitation light in the ALA-PDD step, a known light source can be used, and examples thereof include a purple LED, preferably a flashlight type purple LED, and a light source such as a semiconductor laser. A purple LED, which makes the device compact and is advantageous in terms of cost and portability, particularly a flashlight type purple LED and a purple semiconductor diode can be preferably exemplified. Further, in order to detect the PpIX accumulation site and to detect the fluorescence of red, specifically, the fluorescence having a wavelength of 610 nm to 750 nm, preferably 625 to 638 nm, for determining the propagation range of the periodontal disease pathogen to be irradiated. Examples of the red fluorescence detection device include a detection device with the naked eye and a detection device with a CCD camera.

Examples of the ALA-PDD device in which the excitation light irradiation device and the red fluorescence detection device are integrated include a light source and a small-diameter optical fiber for measurement, and the excitation to be irradiated in the ALA-PDD step for exciting the accumulated PpIX. As a light source, a semiconductor having a strong radiation illuminance to enable detection of PpIX even for a small propagation site of periodontal disease-causing bacteria and a narrow irradiation area to enable accurate automatic identification. A laser light source is preferable, and it is preferable to have an excitation light light guide unit that guides the excitation light and emits it to the outside from one end. Specific examples of the excitation light light guide unit include a small-diameter optical fiber. As an element used as a light source, a semiconductor mixed crystal such as InGaN can be used, and purple light can be oscillated by changing the blending ratio of InGaN. Specifically, a compact laser diode having a diameter of about 5.6 mm can be preferably exemplified. A device in which the port for 4 laser outputs from the laser diode and the port for spectrum measurement are about the size of a desktop PC connected by a built-in high-sensitivity spectrometer can be exemplified. Further, in the light receiving step of receiving the fluorescence emitted by PpIX excited by the excitation light, a small-diameter optical fiber for measurement is used, and the small-diameter optical fiber for measurement is integrated with the small-diameter optical fiber for a light source, and the received fluorescence is received. Is guided to the detector to determine the PpIX accumulation site.

Hereinafter, the present invention will be specifically described with reference to examples, but the technical scope of the present invention is not limited to these examples.

E. Faecalis JCM5803 brain heart infusion medium (BHI) (Difco Laboratories, Detroit , Mich, USA) at, for 8 hours at 37 ° C., and adjusted to 0.8 × 10 ⁹ CFU / ml. Then, under anaerobic conditions at _{37 ℃ (N 2 80%,} CO 2 10%, H 2 10%) was allowed to stand 6 hours to obtain a bacterial cell sample was adjusted to 1.0 × 10 ⁹ CFU / ml .. ALA was dissolved in 1.2 ml of PBS (nacalai tesque, Tokyo), and 0.3 ml of 10N NaOH solution was further added to prepare an ALA solution having a pH of 5.0. The above-adjusted E.I. Fecalis cells and the above-adjusted ALA solution were added to adjust the final concentrations of ALA to 0% (w / v), 0.05% (w / v), 0.5% (w / v), 5% (w / w / v), adjusted to 10% (w / v). Stand in a dark place under anaerobic conditions at 37 ° C., and after 2 hours, a blue LED (wavelength 397 ± 13 nm, output 35 mW, irradiation diameter 15.5 mm, average power density 19 mW / cm ² , G-Light Prima-II Plus, GC, Tokyo) was irradiated on the optical stage for 1 minute. Based on the dilution plate method, the culture broth obtained by irradiation with light was diluted with BHI so that OD = 0.1, and each diluted culture broth was cultured on an agar medium for 1 day. The result is shown in FIG. For those adjusted to the final concentration of ALA of 0% (w / v), a group in which light irradiation by the blue LED was not performed was also provided.
The colonies on each agar medium in FIG. 1 had ALA concentrations of 0% (w / v), 0.05% (w / v), and 0.5% (w / v) from the top for each row. It represents colonies (1 to 6 in FIG. 1) formed from a culture medium of 5, 5% (w / v), 10% (w / v), 0% (w / v) + non-irradiated culture medium.

As is clear from FIG. 1, when the final concentration of ALA is 5% (w / v) and 10% (w / v), the E.I. It was suggested that the viable cell count of Fecalis was significantly reduced, and that the therapeutic agent of the present invention could be used to treat periodontal disease without invading the human body.

[Reference example 1]
E. Faecalis JCM5803 brain heart infusion medium (BHI) (Difco Laboratories, Detroit , Mich, USA) at, for 8 hours at 37 ° C., and adjusted to 0.8 × 10 ⁹ CFU / ml. ALA was dissolved in PBS (nacalai tesque, Tokyo) in 1.2 ml, and 0.3 ml of 10N NaOH solution was added to prepare an ALA solution having a pH of 5.0. An ALA-EDTA solution was prepared by adding EDTA to the above-prepared ALA solution so as to have a concentration of 0.05% (w / v). The above-adjusted E.I. Fecalis cells and the above-adjusted ALA solution or ALA + EDTA solution were added to adjust the final concentrations of ALA to 0% (w / v), 0.05% (w / v), and 0.5% (w / v), respectively. .. After 2 hours and 4 hours, the blue LED (wavelength 397 ± 13 nm, output 35 mW, irradiation diameter 15.5 mm, average power density 19 mW / cm ² , G-Light Prima) was allowed to stand in a dark place at 37 ° C. under aerobic conditions. -II Plus, GC, Tokyo) was irradiated on the optical stage for 1 minute. Based on the dilution plate method, the culture solution obtained by irradiation with light was diluted 1-fold, 10-fold, 100-fold, and 1000-fold with BHI, and each diluted culture solution was cultured on an agar medium for 1 day. The result is shown in FIG.
The results of 1-fold dilution, 10-fold dilution, 100-fold dilution, and 1000-fold dilution of the culture solution are shown from the left in FIG. 2, and the colonies on each agar medium are the culture solutions to which the ALA solution is added from the top for each row. Colonies formed from cultures having ALA concentrations of 0% (w / v), 0.05% (w / v), and 0.5% (w / v) (1 to 3 in FIG. 2). ), The ALA concentration in the culture solution to which the ALA + EDTA solution was added was formed from the culture solution of 0% (w / v), 0.05% (w / v), 0.5% (w / v). Represents a colony (4-6 in FIG. 2).

As is clear from FIG. 2, when the cells were cultured under aerobic conditions and irradiated with light, E.I. The viable cell count of Fecalis was completely the same even when ALA was added as in the case of blank, and the effect of adding ALA was not observed.

The therapeutic agent and the therapeutic method of the present invention are useful in the field of treating periodontal disease because they are effective against periodontal disease-causing bacteria that have not been effective by the conventional methods.

Claims (6)

The following formula (I)

(In the formula, R ¹ represents a hydrogen atom or an acyl group, and R ² represents a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group or an aralkyl group) or a pharmaceutically acceptable compound. 5-Aminolevulinic acid, which contains the salt of hydrogen, is administered to the affected area of periodontal disease with the compound represented by the formula (I) or the pharmaceutically acceptable salt thereof, and then irradiated with light having a wavelength of 360 nm to 700 nm. A therapeutic agent for periodontal disease for photodynamic therapy (ALA-PDT). The therapeutic agent according to claim 1, wherein the periodontal disease-causing bacterium in periodontal disease is Enterococcus faecalis. The therapeutic agent according to claim 1, wherein the light having a wavelength of 360 nm to 700 nm is a blue light emitting diode. A method for treating periodontal disease, in which a compound represented by the above formula (I) or a pharmaceutically acceptable salt thereof is administered to the affected part of periodontal disease, and then light having a wavelength of 360 nm to 700 nm is irradiated. A compound represented by the above formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of periodontal disease, which is administered to the affected area of periodontal disease and then irradiated with light having a wavelength of 360 nm to 700 nm. Compounds represented by the above formula (I) or pharmaceutically acceptable compounds for use in the production of therapeutic agents for periodontal disease, which are administered to the affected area of periodontal disease and then irradiated with light having a wavelength of 360 nm to 700 nm. Use of salt.

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