JP2009263317A - Therapeutic agent for skin disease - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a therapeutic agent for skin disease using PDT (Photodynamic Therapy). <P>SOLUTION: A chlorin derivative is shown by formula (I), wherein X is O or S, Y is -(CH<SB>2</SB>)<SB>n</SB>- etc., and R is -OH, -OCH<SB>3</SB>, etc. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a skin disease therapeutic agent used for photophysical chemical diagnosis / treatment (PDT: Photodynamic Therapy) comprising a novel chlorin derivative or a pharmacologically acceptable salt thereof as an active ingredient.

  As a new treatment method for cancer, a photophysicochemical diagnosis / treatment method (hereinafter referred to as “photodynamic therapy”: PDT: Photodynamic Therapy) is performed. In this method, a certain porphyrin derivative is administered by a method such as intravenous injection and selectively accumulated in cancer (tumor) tissue, and then only the cancer tissue is selectively destroyed by irradiation with laser light. Is a treatment method that uses two characteristics of selective accumulation in a cancer tissue and a photosensitizing action of a porphyrin derivative.

The present inventors have been diligently researching a porphyrin derivative that can be used for this PDT, and have provided an iminochlorin aspartic acid derivative referred to as ATX-S10.
Among the iminochlorin aspartic acid derivatives or pharmacologically acceptable salts thereof, particularly a compound that is a sodium salt, that is, a compound that is named ATX-S10 · Na, has an ability to accumulate in cancer tissue, as well as neovascularization. It is a compound that has a remarkably high selective accumulation property. Therefore, it has been confirmed that it is extremely effective as a therapeutic agent for PDT for the treatment of tumors using its excellent characteristics (Patent Document 1).

  On the other hand, inflammatory keratosis, which is caused by inflammation caused by new blood vessels, is caused by the spread of blood vessels in the dermis of the skin and the infiltration of white blood cells such as lymphocytes into the skin and the skin epidermis. Is a skin symptom in which "keratosis" that causes thickening of the stratum corneum and thickening of the stratum corneum occurs at the same time. Anti-inflammatory agents such as steroids, epidermal growth inhibitors such as retinoids, and UV therapy (PUVA therapy) are implemented. However, it is not a definitive treatment leading to complete cure. In recent years, it has been found that treatment with PDT using 5-aminolevulinic acid hydrochloride (5-ALA) is effective. However, 5-ALA is a biosynthetic precursor of protoporphyrin IX, and the compound properties of protoporphyrin IX, that is, the low neovascularization, and the longest wavelength absorption edge are 630 nm, provide a sufficient therapeutic effect. I can't.

  One of the keratosis is actinic keratosis. This actinic keratosis is also called senile keratosis and is said to be a “cancer precursor” because it may progress to skin cancer in the future. It is a keratosis caused by long-term exposure to sunlight that damages the nuclear DNA DNA of skin cells and makes it difficult to repair with age. Symptoms are crisp, slightly raised erythema, or gray or brown pigment spots that appear on the skin surface and seem to resemble eczema or warts. It is preferable to treat and excise. Treatment with surgery, excision with a carbon dioxide laser, cryotherapy, anticancer agent-containing ointment, etc. are carried out, but treatment with PDT is also effective.

  By the way, ATX-S10 · Na proposed by the present inventors as a result of the study by the present inventors is also excellent in the accumulation of inflammatory cells caused by new blood vessels other than cancer and ophthalmology, and applied PDT. As a skin disease therapeutic agent, it was proved to be effective as a therapeutic agent for inflammatory keratosis (dermatitis such as psoriasis), and as a skin disease therapeutic agent applying PTX as ATX-S10 · Na A patent application has already been filed for this point (Patent Document 2).

However, the ointment containing ATX-S10 · Na has low skin permeability, and the stratum corneum must be removed by tape stripping in order to reach the effective therapeutic amount in the affected area.
International Publication WO 98/14453 Japanese Patent Laid-Open No. 2004-026717

In view of the above-described present situation, the present inventors have studied to develop a skin disease therapeutic agent for PDT that does not require tape stripping and reduces the burden on the patient. That is, what is effective as a skin disease treatment agent used in PDT therapy requires that the drug itself has high skin permeability and that it is fat-soluble so that it can be uniformly dissolved and dispersed in the base as an ointment. Is done. Based on this viewpoint, several chlorin derivatives were synthesized and examined. As a result, the novel chlorin derivative represented by the formula (I) described later was found to be extremely effective, and the present invention was completed. I came to let you.
Therefore, an object of the present invention is to provide a skin disease therapeutic agent using PDT (Photodynamic Therapy).

  The present invention for solving such a problem has the following formula (I):

(Where
X represents O or S;
Y represents — (CH ₂ ) _n — or — (CHOH) —,
R represents —OH, —O (CH ₂ ) _m CH ₃ —, —O (CH ₂ ) ₁ —OH or —S (CH ₂ ) ₁ —SH;
n represents an integer of 0 to 10,
m represents an integer of 0 to 11,
l represents an integer of 2 to 12)
A dermatological treatment agent for photodynamic therapy (PDT) comprising a chlorin derivative represented by the formula (1) or a pharmacologically acceptable salt thereof as an active ingredient.

  Among them, the present invention particularly relates to the following formula (Ia):

(In the formula, X represents O or S, Y represents — (CH ₂ ) _n —, and n is an integer of 0 to 4).
Or a pharmacologically acceptable salt thereof is preferred.

  Specifically, the present invention provides photodynamics comprising 0.1 to 20.0% by weight of the chlorin derivative represented by the above formula (I) or a pharmacologically acceptable salt thereof with respect to the formulation weight. It is an ointment for treating skin diseases for therapy (PDT).

  More specifically, in the present invention, the ointment base is FAPG (H) ointment base, FAPG (K) ointment base, PEG ointment base, PEG-PG ointment base, petrolatum ointment base, SR petrolatum ointment. A skin disease treatment ointment for photodynamic therapy (PDT) that is selected from a base, SR petrolatum-IPM ointment base or plastibase ointment base.

  That is, the basic aspect of the present invention is characterized in that skin diseases are treated with PDT using the chlorin derivative represented by the above formula (I) or a pharmacologically acceptable salt thereof.

The ointment containing the chlorin derivative represented by the above formula (I) or a pharmacologically acceptable salt thereof provided by the present invention does not require tape stripping and has good permeability to the affected skin area. Yes, and therefore does not burden the patient in PDT therapy in the treatment of skin diseases.
Further, since tape stripping is not required, the treatment itself has an advantage that it can be easily performed, and a new treatment system can be provided.

As described above, the active ingredient in the skin disease therapeutic agent by PDT therapy provided by the present invention is a chlorin derivative represented by the formula (I) or a pharmacologically acceptable salt thereof (hereinafter simply referred to as “chlorin derivative”). It is.
This chlorin derivative is different from ATX-S10, which is an iminochlorin aspartic acid derivative that has been provided by the present inventors, so that it has high skin permeability and lipid solubility. And the stability of the ointment preparation itself is very good.

  Among such chlorin derivatives represented by the formula (I), the following formula (Ia):

Or a pharmacologically acceptable salt thereof, specifically, a compound represented by the following abbreviation.
PG-P-Me: a chlorin derivative in which X is O and Y is —CH ₂ — in formula (Ia),
EG-P-Me: a chlorin derivative in which in formula (Ia), X is O and Y is a direct bond,
ET-P-Me: A chlorin derivative in which X is S and Y is a direct bond in formula (Ia).

These chlorin derivatives can be obtained, for example, as follows.
That is, it is prepared by dissolving a corresponding photoprotoporphyrin derivative in a suitable organic solvent and reacting with a diol compound such as 1,3-propanediol, ethylene glycol, 1,2-ethanedithiol or a dithiol compound. can do.

  The organic solvent used for the reaction is not particularly limited and can be arbitrarily selected as long as it does not directly affect the reaction. Specific examples include aromatic hydrocarbons such as benzene, toluene and xylene, and ether solvents such as ether, tetrahydrofuran and dioxane. Of these, tetrahydrofuran is preferably used.

The reaction temperature and reaction time are also not particularly limited, and the stirring treatment is preferably performed at 0 to 60 ° C., preferably at room temperature for about 0.5 to 10 hours.
The reaction requires the presence of a mild acid catalyst, and examples of such an acid catalyst include methanesulfonic acid, toluenesulfonic acid, p-toluenesulfonic acid and the like, and among them, p-toluene. By using sulfonic acid, the target chlorin derivative can be obtained in high yield.

Specific examples of some of the preparation methods are shown below.
Production Example 1: Preparation of a chlorin derivative in which X is O and Y is —CH ₂ — in the formula (Ia) (Preparation of PG-PM)
1.0 g of photoprotoporphyrin was dissolved in 50 mL of tetrahydrofuran, and 4.8 mL of 1,3-propanediol (40 times equivalent) and 150 mg of p-toluenesulfonic acid monohydrate (0.5 times equivalent) were added to this solution. Was added and stirred for 2 hours. After completion of stirring, 1% aqueous sodium hydrogen carbonate solution was added and the deposited precipitate was collected by filtration and purified by a silica gel column to obtain 0.9 g of a new chlorin derivative (referred to as PG-P-Me). When the obtained derivative was subjected to mass spectrometry by LC-MS (electrospray, ESI), it showed m / z: 681 and was confirmed to be the intended chlorin derivative.

Production Example 2: Preparation of chlorin derivative in which X is O and Y is a direct bond in formula (Ia) (Preparation of EG-P-Me)
Dissolve 1.0 g of photoprotoporphyrin in 50 mL of tetrahydrofuran, and add 4.0 mL (40 times equivalent) of ethylene glycol and 150 mg (0.5 times equivalent) of p-toluenesulfonic acid monohydrate to the solution. By the same treatment as in Example 1, 0.9 g of a novel chlorin derivative (referred to as EG-P-Me) was obtained. When the obtained derivative was subjected to mass spectrometry by LC-MS (electrospray, ESI), m / z: 667 was shown, which was confirmed to be the intended chlorin derivative.

Production Example 3: Preparation of a chlorin derivative in which X is S and Y is a direct bond in the formula (Ia) (Preparation of ET-P-Me)
1.0 g of photoprotoporphyrin is dissolved in 50 mL of tetrahydrofuran, and 6.0 mL of 1,2-ethanedithiol (40 times equivalent) and 150 mg of p-toluenesulfonic acid monohydrate (0.5 times equivalent) are added to this solution. And 0.4 g of novel chlorin derivative (referred to as ET-P-Me) was obtained by the same treatment as in Production Example 1. When the obtained derivative was subjected to mass spectrometry by LC-MS (electrospray, ESI), it showed m / z: 699, which was confirmed to be the intended chlorin derivative.

In the present invention, these chlorin derivatives are formulated as a skin disease treatment ointment for PDT. Various bases can be used as the ointment base, and the formulation of an ointment preparation that can ensure the stability of the preparation over time was examined. As a result, it was found that a non-aqueous ointment base could be a very good ointment.
Specific examples of such non-aqueous ointment bases include FAPG (H) ointment base, FAPG (K) ointment base, PEG ointment base (macrogol ointment base), and PEG-PG ointment base. , Petrolatum ointment base, SR petrolatum ointment base, SR petrolatum-IPM ointment base, plastibase ointment base.

  Specifically, the FAPG (H) ointment base is an ointment base made of stearyl alcohol, stearic acid and propylene glycol. The FAPG (K) ointment base is an ointment base made of stearyl alcohol, stearic acid and polyethylene glycol 400. The PEG ointment base (macrogol ointment base) is an ointment base composed of polyethylene glycol 400 and polyethylene glycol 4000, and the PEG-PG ointment base is an ointment base composed of propylene glycol and polyethylene glycol 4000.

  Vaseline ointment base is an ointment base composed of petrolatum and liquid paraffin, and SR petrolatum ointment base is an ointment base composed of SR petrolatum (5% salicylic acid-petroleum) and SR petrolatum- The IPM ointment base is an ointment base composed of SR petrolatum and isopropyl myristate. Furthermore, the plastibase ointment base is an ointment base made of plastibase (95% liquid paraffin and 5% polyethylene resin).

It should be noted that the ointment base used in the present invention is not limited to those described above, and it is needless to say that an ointment base generally used in pharmacology can be used. It is also acceptable to add other stabilizers pharmacologically or pharmacologically and transdermal absorption enhancers such as l-menthol.
According to the study by the present inventors, it has been found that the macrogol ointment base is particularly preferable among the above-described ointment bases.

  In the present invention, the amount of the chlorin derivative represented by the formula (I) to be contained in these ointment bases is such that the chlorin derivative, which is the compounded active ingredient, is percutaneously absorbed and accumulated at the diseased site, and irradiated with a laser beam. It is sufficient that an amount sufficient to kill the target cell is added. According to the study by the present inventors, it was found that a sufficient effect can be obtained if the blending amount is 0.1 to 20% by weight based on the weight of the preparation.

When the blending amount is less than 0.1% by weight, the intended therapeutic effect cannot be increased, and even when blended in an amount of 20.0% by weight or more, no further effect can be obtained.
The blending amount cannot be generally specified depending on the type of active ingredient to be contained, and the stability of the active ingredient to be contained is not greatly affected by the concentration of the active ingredient and the ointment base to be used. Within the range, it is possible to make various changes according to the application.

  In order to produce the ointment preparation of the present invention, a widely used method for preparing an ointment preparation is used, and a chlorin derivative, which is an active ingredient, may be uniformly kneaded in an ointment base.

When the ointment preparation of the present invention obtained as described above is used for PDT, it is effective by directly applying to a skin disease site such as actinic keratosis, inflammatory keratosis, epidermis cancer, infection, etc. Then, the chlorin derivative is accumulated, and then the disease can be effectively treated by irradiating the site with light such as a laser beam.
In application of this ointment, the chlorin derivative provided by the present invention does not require tape stripping and has good permeability to the affected skin area, and therefore imposes a burden on patients in PDT in the treatment of skin diseases. This has the advantage that the treatment itself can be easily performed.
In applying the ointment, it is more effective to keep the application site sealed in order to obtain an ODT effect (occlusive dressing technique).

  Various laser beams and lamps can be used for light irradiation at a diseased site after applying an ointment containing a chlorin derivative provided by the present invention. For example, a titanium sapphire laser, a semiconductor laser, an OPO-YAG laser, a xenon lamp, a halogen lamp, a metal halide lamp, a light emitting diode, or the like can be used. Among them, it is more effective to use a semiconductor laser for a chlorin derivative having an absorption wavelength near 660 to 670 nm.

  Thus, the disease can be effectively treated by applying the ointment preparation of the present invention to the diseased site and absorbing the active ingredient percutaneously and then irradiating the diseased site with light.

  Hereinafter, the present invention will be described in detail with specific formulation examples, test examples, and the like, but the present invention is not limited to these examples.

Specific formulation examples of ointments containing the chlorin derivative of formula (I) provided by the present invention are described below.
Formulation Example 1: Preparation and stability test of PEG ointment base (macrogol ointment) [Preparation of ointment]
Polyethylene glycol 4000 (PEG 4000, average molecular weight of about 3,000; manufactured by Wako Pure Chemical Industries, Ltd.) 3.0 g, polyethylene glycol 400 (PEG 400, average molecular weight of about 360-440; manufactured by Wako Pure Chemical Industries, Ltd.) 6.6 g and l- Menthol (0.1 g) was heated and dissolved and mixed, and 0.3 g of the new chlorin derivative (PG-P-Me) obtained in Production Example 1 was added and mixed there. The mixture was allowed to cool with stirring to prepare the intended macrogol ointment of the present invention.

[Stability test]
The macrogol ointment obtained above was stored in a refrigerator for 3 months in a polypropylene container, and the purity of the active ingredient (PG-P-Me) was measured by HPLC to evaluate its stability. Purity before the start of storage and purity after storage for 3 months were determined by area percentage by HPLC.
As a result, it was found that the purity of the active ingredient (PG-P-Me) was not observed and was stable.

  Next, the actual treatment effect of PDT on the ointment containing the novel chlorin derivative provided by the present invention will be described.

Test Example 1: Preparation of a papilloma (papilloma) model mouse 0.2 μM 7,12-dimethylbenzanthracene (DMBA) was applied once to the back of a 12-week-old female hairless mouse (HOS: Hr Shizuoka Laboratory Center). After 2 weeks, 0.02 μM 12-O-tetradecanoyl phorbol-13-acetate (TPA) was applied twice a week.
As a result, many papillomas having a diameter of 1 to 2 mm appeared after 2 months.

Test Example 2: Treatment effect of ointment using papilloma model mouse by PDT therapy [Method]
Macrogol ointment (containing PG-P-Me) prepared in Formulation Example 1 was applied to the papilloma portion of the papilloma model mouse prepared in Test Example 1 to a large extent, and the application site was kept sealed for 4 hours. Thereafter, the ointment was wiped off.
The papilloma part from which the ointment was removed was irradiated with a laser using a semiconductor laser (150 mW / cm ² , 670 nm, LD670-05; manufactured by Hamamatsu Photonics).
After irradiation, the naked eye was appropriately observed.

[result]
The results are shown in FIGS.
Each figure is a photograph of a papilloma (papilloma) portion of a papilloma model mouse.
1 and 2 are photographs showing the results of control in which laser irradiation (200 J / cm ² ) was performed without applying the ointment of the present invention.
FIG. 1 is a photograph before irradiation, and FIG. 2 is a photograph two days after irradiation (48 hours). In the control to which the ointment of the present invention was not applied, disappearance of papilloma was not confirmed.

3 to 6 are photographs showing the results of applying an ointment and irradiating with laser (200 J / cm ² ).
FIG. 3 is a photograph before irradiation. FIG. 4 shows the results after 5 hours of irradiation, FIG. 5 after 3 days of irradiation, and FIG. 6 after 7 days of irradiation.
By applying the ointment for treatment of skin diseases of the present invention and performing PDT therapy, papilloma shrinkage was observed even after 5 hours of irradiation, and papilloma completely disappeared after 7 days of irradiation.

7 to 9 are photographs showing the results of applying an ointment and irradiating with laser (100 J / cm ² ).
FIG. 7 is a photograph before irradiation. FIG. 8 shows the result after 3 hours of irradiation, and FIG. 9 shows the result after 24 hours of irradiation.
By applying the skin disease treatment ointment of the present invention and performing PDT therapy, a significant reduction in papilloma was observed 24 hours after irradiation.

10 to 14 are photographs showing the results of applying an ointment and irradiating with laser (50 J / cm ² ).
FIG. 10 is a photograph before irradiation. FIG. 11 shows the results after 3 hours of irradiation, FIG. 12 after 24 hours of irradiation, FIG. 13 after 48 hours of irradiation, and FIG. 14 after 6 days of irradiation.
By applying the ointment for treating skin diseases of the present invention and performing PDT therapy, a significant reduction in papilloma was observed over time, and the papilloma disappeared completely 6 days after irradiation.

  As described above, the present invention provides a chlorin derivative for use as photodynamic therapy (PDT), and the chlorin derivative provided by the present invention can be used as a skin-applied ointment preparation. (Percutaneous absorbability) is very good. Therefore, by applying to skin diseases such as actinic keratosis, inflammatory keratosis, epidermis cancer, infectious diseases, such as papilloma, chlorin derivatives are effectively accumulated at the diseased site, and laser irradiation Can effectively treat skin diseases, and its medical value is tremendous

6 is a photograph of a control before laser irradiation in Test Example 2. 4 is a control photograph 48 hours after laser irradiation in Test Example 2. 6 is a photograph before laser irradiation (200 J / cm ² ) in Test Example 2. It is the photograph in the test example 2 after laser irradiation (200 J / cm < ² >) 5 hours. 3 is a photograph 3 days after laser irradiation (200 J / cm ² ) in Test Example 2. 7 is a photograph 7 days after laser irradiation (200 J / cm ² ) in Test Example 2.

4 is a photograph before laser irradiation (100 J / cm ² ) in Test Example 2. It is a photograph in laser irradiation (100 J / cm ² ) 3 hours after Test Example 2. It is a photograph in laser irradiation (100 J / cm ² ) 24 hours after Test Example 2.

6 is a photograph before laser irradiation (50 J / cm ² ) in Test Example 2. It is a photograph in laser irradiation (50 J / cm ² ) 3 hours after Test Example 2. It is a photograph in laser irradiation (50 J / cm ² ) 24 hours after Test Example 2. 4 is a photograph 48 hours after laser irradiation (50 J / cm ² ) in Test Example 2. 6 is a photograph 6 days after laser irradiation (50 J / cm ² ) in Test Example 2.

Claims (3)

Formula (I):

(Where
X represents O or S;
Y represents — (CH ₂ ) _n — or — (CHOH) —,
R represents —OH, —O (CH ₂ ) _m CH ₃ —, —O (CH ₂ ) ₁ —OH or —S (CH ₂ ) ₁ —SH;
n represents an integer of 0 to 10,
m represents an integer of 0 to 11,
l represents an integer of 2 to 12)
A dermatological treatment agent for photodynamic therapy (PDT) comprising a chlorin derivative represented by the formula (I) or a pharmacologically acceptable salt thereof as an active ingredient.   The photodynamic therapy (PDT) according to claim 1, wherein the chlorin derivative represented by the formula (I) or a pharmacologically acceptable salt thereof is contained in an amount of 0.1 to 20.0% by weight based on the weight of the preparation. Ointment for the treatment of skin diseases.   Ointment base is FAPG (H) ointment base, FAPG (K) ointment base, PEG ointment base, PEG-PG ointment base, petrolatum ointment base, SR petrolatum ointment base, SR petrolatum-IPM ointment base The ointment for skin disease treatment for photodynamic therapy (PDT) according to claim 4, wherein the agent is selected from an agent or a plastibase ointment base.

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