KR101308507B1 - Acne therapeutics and sebum secernent inhibitor which comprise tryptophan, and kits for photodynamic therapy containing the same - Google Patents

Abstract

The present invention relates to novel uses of Tryptophan acne as a therapeutic and sebum secretion inhibitor. More specifically, the present invention relates to a photosensitizer for photodynamic therapy (PDT) containing tryptophan having properties activated by light, and a kit for photodynamic therapy and a composition for photodynamic therapy comprising the same It relates to a novel use as a therapeutic agent for acne and as an inhibitor of sebum secretion. Ultraviolet light or visible light may be used as a light source for activating tryptophan, and it is particularly preferable to use blue light or green light.

Description

ACNE THERAPEUTICS AND SEBUM SECERNENT INHIBITOR WHICH COMPRISE TRYPTOPHAN, AND KITS FOR PHOTODYNAMIC THERAPY CONTAINING THE SAME}

The present invention relates to novel use of Tryptophan and its derivatives as a therapeutic agent for acne and as an inhibitor of sebum secretion.

More specifically, the present invention provides a photosensitizer for photodynamic therapy (PDT) containing tryptophan having a property activated by light, and a kit for photodynamic therapy and a composition for photodynamic therapy comprising the same It relates to a novel use as a therapeutic agent for acne and as an inhibitor of sebum secretion.

Ultraviolet light or visible light may be used as the light source for activating the tryptophan of the present invention, and it is particularly preferable to use blue light or green light.

Photodynamic therapy (PDT) is one of the most promising methods of treatment in recent years. The principle of photodynamic therapy is the single-state oxygen derived from a comprehensive chemical reaction between oxygen, externally supplied light (photons) and light-sensitive substances (photosensitizers). Or free radicals are a method of treating cancer by destroying various lesion sites or cancer cells.

The advantage of this photodynamic therapy is that the diseased cells can be selectively removed while preserving normal cells, thereby eliminating the risk of general anesthesia, and the procedure is easy because the operation can be performed simply by local anesthesia. .

Therefore, photodynamic therapy is not only superior to the specific effect of removing cancer cells, but also can be quickly recovered after the procedure and shortened the hospitalization period, thereby improving the quality of life of patients and facilitating the return to social activities. That has the advantage.

Photodynamic therapy has been studied since the 1980s, and since the clinical procedure was approved in Canada, Germany and Japan in the 1990s, the US FDA approved the treatment of esophageal cancer in January 1996. It has been approved for the treatment of lung cancer. In early 1996, 3,000 photodynamic treatments were performed in approximately 32 countries.

However, the current photodynamic therapy is a problem that the use of the photodynamic therapy in the bulky tumor is limited because the light can not penetrate the entire tumor, the porphyrin represented by most of the photosensitizer is expensive Side effects of phototoxicity occur due to slow metabolism, and there is a problem that the effect of photodynamic therapy cannot be sufficiently seen because the concentration of photosensitizer in the tumor is low.

Photofrin ^™, a photosensitizer used in photodynamic therapy, was approved by the US FDA in 1996 and has a good therapeutic effect and stability. However, it can accumulate in the body for 5 to 6 weeks after administration and cause side effects, and pure Photofrin ^TM is difficult to manufacture and due to absorption at 630 nm, lower than 650-850 nm, which is known to be optimal for photodynamic therapy. Only a few millimeters can penetrate the cell, which has the disadvantage of relatively low therapeutic effect (Chemistry & Industry, Sep 21, 1998, 739-743: Chemical & Engineering News, Nov 2, 1998, 22-27). Therefore, there is an urgent need for the development of new photosensitizers that overcome these problems.

Porphyrins, chlorines, bacteriochlrins, and porphycenes are the most studied compounds known as next-generation photosensitizers (J Org. Chem., 63, 1998, 1646). -1656). Among these, the phephyphytins, which are metal ions removed from chlorophylls, which are widely distributed in plants, are not only absorbed at longer wavelengths than Photofrin ^TM , a hematoporphyrin derivative, but also separated from the high purity. As much as possible, a lot of research has been focused on the next generation of photosensitizers, but the results have not been obtained.

In recent years, attempts have been made to use such photodynamic therapy to treat acne and psoriasis. Aminolevulinic acid (ALA) is mainly used as a light sensitizer for acne treatment. Like this photosensitizer, this drug also needs to avoid light for a long time. The same side effects are severe.

As mentioned above, there have been various researches and efforts on photo sensitizers to date, and some of them have been developed as acne treatments, but there are problems of patients' discomfort in daily life and serious side effects. Since it has not been harvested, there is an urgent need in the art for the development of new photosensitizers for simple and safe treatment and effective application to the human body.

On the other hand, the present inventors focused on the fact that it is not known that tryptophan is effective in treating acne and inhibiting sebum secretion until now, and as a result of intensive efforts, it has been shown that in recent years, when tryptophan is used in combination with light, it has excellent anti-acne effect. The present invention was completed by confirming that it can be used as a treatment for acne and also has a therapeutic effect as an inhibitor of sebum secretion.

The basic object of the present invention is to provide a novel use of tryptophan as a therapeutic agent for acne photodynamics.

Another object of the present invention is to provide a novel photodynamic therapeutic agent as an inhibitor of sebum secretion of tryptophan.

It is still another object of the present invention to provide a kit for photodynamic therapy and a pharmaceutical composition for photodynamic therapy comprising the tryptophan photosensitizer for using tryptophan as an acne treatment or sebum secretion inhibitor.

The basic object of the present invention described above can be achieved by identifying a novel pharmacological effect as Tryptophan as a photosensitizer for treating acne or inhibiting sebum secretion.

Still another object of the present invention is a photosensitizer for photodynamic therapy (PDT) containing tryptophan having a property of being activated by light to treat acne or inhibit sebum secretion and a kit for photodynamic therapy comprising the same And by providing a composition for photodynamic therapy.

Ultraviolet light or visible light may be used as the light source for activating the tryptophan of the present invention, and it is particularly preferable to use blue light or green light.

In order to achieve the above objects, the present invention provides a photosensitizer containing tryptophan of Formula 1 and its derivatives for acne treatment or sebum secretion control.

            Formula 1

The present invention also provides a composition for treating photodynamics containing a therapeutically effective amount of tryptophan having the structure of Formula 1 having photoactivation properties as an active ingredient for acne treatment and sebum secretion control.

In another aspect, the present invention provides a kit for photodynamic therapy comprising a tryptophan having the structure of Formula 1 and a light source for in vivo or ex vivo light irradiation.

Hereinafter, the present invention will be described in more detail.

Tryptophan having the structure of Chemical Formula 1 does not require the use of a separate photosensitizer and acts as a photosensitizer itself, which is photoactivated by light irradiation and sterilization of skin bacteria such as acne and staphylococci. It is characterized by the effect.

Light rays capable of photoactivating tryptophan are light rays in the ultraviolet and visible range, and there is no limitation in the wavelength range. However, in the case of photoactivating tryptophan in vivo, it is preferable to use light of 280 nm or more in consideration of adverse effects on normal tissues, and the longer the wavelength, the lower the energy and the longer the irradiation time. It is preferable to use light rays in the range of 280 nm to 1000 nm, and more preferably, light rays in the range of 300 nm to 750 nm.

In addition, considering the photoactivation efficiency of tryptophan, it is preferable to use long wavelength ultraviolet light between 350 and 400 nm, blue light between 400 and 500 nm, or green light between 500 and 600 nm. In addition, in consideration of the degree of tryptophan activation, the degree of cell penetration and biosafety, blue light or green light is most preferred.

The light source for irradiation of the light beam is selected from the group consisting of an ultrasonic irradiation emitter, a light emitting diode, a laser diode, a dye laser, a metal halide lamp, a flash lamp, a mechanically filtered fluorescent light source, a mechanically filtered incandescent light, and a filament light source. It may be one or more selected from the group consisting of a light source for in vitro light irradiation.

When photoactivating tryptophan by light rays, tryptophan can be irradiated and exposed photoactively for a therapeutically effective pulse duration to light of one or more wavelengths irradiated from the light source.

When photoactivating tryptophan, there is no limit to the intensity (energy) of the light that is irradiated, the lower the intensity of the light, the longer the duration and / or increase the number of irradiation, and the higher the intensity, the shorter the duration. And / or reduce the number of irradiation to photoactivate the tryptophan.

When tryptophan is photoactivated, if the light intensity is too low, it may penetrate into the target tissue appropriately to exhibit an effective tryptophan photoactivation effect, and if the light intensity is too high, side effects such as necrosis of normal tissue may occur. Thus, the intensity of the irradiated light is preferably in the range of 1 to 100 J / cm ² .

In addition, if the pulse duration is too short or the number of irradiation is too small, the light activation effect is insignificant, and if the pulse duration is too long or the irradiation time is too large, side effects such as necrosis of normal tissue may appear.

In addition, as described above, depending on the intensity of the light to be irradiated, when the light intensity is low, the duration or increase the number of irradiation, when the light intensity is high, the duration is short or the number of irradiation is safe and useful effect Can be seen.

Therefore, the pulse duration and the number of irradiation can be appropriately adjusted in consideration of the light intensity range and adverse effects on normal tissue, etc., but it is difficult to define uniformly, but preferably the pulse duration is 0.1 to 500 ms, The number of irradiation may be effective from 1 to 100 times.

For example, in one embodiment of the present invention, the intensity of light was 20 J / cm ² , the pulse length was 15 ms, and the number of times of irradiation of the light was 1 cycle in the acne treatment experiment conducted in the human body. It was. Preferably, the light was activated by a strong light irradiation device such as IPL (Intense Pulse Light).

The tryptophan-containing photodynamic therapy composition contains tryptophan having photosensitive activity in an amount of 0.001% by weight to 99% by weight, preferably 0.001% by weight to 30% by weight. In order to obtain sufficient photosensitizing and therapeutic effects of tryptophan, the tryptophan content is preferably 0.001% by weight or more.

The compositions can be used in liquid, semisolid, solid or aerosol form, and are formulated in the form of, for example, aqueous or non-aqueous suspensions, solutions, creams, ointments, gels, syrups, suppositories, tablets, capsules or microdrop sprays. Can be used.

Thus, the composition may further comprise excipients necessary for such formulation. The composition may further include one or more adjuvants selected from the group consisting of conventional preservatives, stabilizers, buffers, pH adjusters, sweeteners, flavoring agents, dyes, etc., depending on the requirements of storage and administration conditions, such as application form or route of administration. It may contain.

In addition, the composition may further contain one or more other kinds of drugs having a desired therapeutic effect.

Tryptophan having a photosensitizing activity contained in the composition may be photoactivated by light irradiation in advance before administration, or may be photoactivated by light irradiation by a light source after administration.

The photodynamic therapy composition or photodynamic therapy kit of the present invention is effective in the treatment of acne and in particular can control sebum secretion.

The tryptophan of the present invention is used as a new photosensitizer to exhibit acne treatment and sebum secretion. Since the tryptophan of the present invention is effectively activated by visible light, especially blue light or green light, it exhibits excellent antibacterial effect and sebum secretion control effect, and thus, it is expected to be used as a novel acne therapeutic agent and commercial use in the pharmaceutical industry as a sebum secretion inhibitor.

1 is a graph demonstrating the effect of tryptophan and IPL combination on P. acnes.
2 is a graph demonstrating the effect of the combination of tryptophan and IPL on S. aureus.
3 is a graph demonstrating the effect of tryptophan and IPL combination on acne on the face.
4 is a graph demonstrating the effect of the combination of tryptophan and IPL on sebum of the face.

Example

Hereinafter, the present invention will be described in more detail with reference to the following examples. The following examples are merely to illustrate the invention, but are not intended to limit the scope of the invention to the content of these examples.

Example 1 bactericidal effect of acne and skin bacteria by tryptophan and light

Through this experiment, the present inventors have confirmed that the bactericidal effect of tryptophan appears when used with light and tryptophan alone does not show any toxicity.

P. acnes 706486 and S. aureus W-1-14 were used as strains. Mueller Hinton ∥ broth (Becton, Dickinson Co., Sparks, USA) was used as a liquid medium for culturing the strain. S. aureus colonies were diluted in sterile Phosphate Buffered Saline (PBS, Invitrogen Co., NY, USA) and diluted to a McFarland turbidity of 0.5. The concentration of bacteria at this time is 1ⅹ10 ⁸ / ml. After further diluting the bacterial concentration to 2 × 10 ⁴ / ml, 12 mueller Hinton agar (Becton, Dickinson Co., Sparks, USA) plates were spread evenly with 10 μl of spreader, and dried on a clean bench for 30 minutes. . Tryptophan was diluted in Dulbecco's Phosphate Buffered Saline (DPBS, Invitrogen Co., NY, USA) to 20 mM, 10 mM, 5 mM, 1 mM, 0.5 mM. 4 ml of tryptophan of 0 mM (tryptophan-free DPBS (tryptophan 0 mM), 0.5 mM, 1 mM, 5 mM, 10 mM, and 20 mM were reacted in a medium to which 6 bacterial solutions were applied for 3 minutes. IPL (Ellipse Flex DDD, Denmark) was then irradiated with an Intermittent Pulse Laser (hereinafter referred to as IPL) using an applicator fitted with a filter between 400 nm and 720 nm 10 J / cm ² . The tryptophan was discarded, the medium was washed twice with PBS, incubated for 24 hours at 37 ° C., and the colonies were counted in the medium to which 6 different bacterial solutions were applied, and 0 mM, 0.5 mM, 1 mM, 5 mM 4 ml of tryptophan, 10 mM, and 20 mM were reacted for 4 minutes, and then treated with the same method without irradiation of IPL, and the number of colonies was counted. Colonies were counted by incubation for 24 hours in an anaerobic chamber replaced with nitrogen gas.

After tryptophan treatment in P. acnes and S. aureus , the number of colonies in the group that caused photodynamic reaction with IPL and those without IPL were compared. In the case of P. acnes , the number of colonies was significantly lower in the IPL group than in the IPL group, and this effect was observed at all tryptophan concentrations from 0.5 mM to 20 mM.

Colonies did not grow in the group irradiated with 10 mM tryptophan in IPL. In the group not treated with IPL, the number of colonies was reduced to 80% or less of the control group when tryptophan was treated at 5 mM or more, so tryptophan at 5 mM or more appeared to inhibit the growth of P. acnes by itself (FIG. 1). .

In the case of S. aureus, the number of colonies was significantly reduced in the IPL group compared to the group without IPL. Colonies did not grow in the group irradiated with 20 mM tryptophan IPL. Also, As tryptophan concentration increased, the number of colonies growing after IPL irradiation decreased. In the group not treated with IPL, the number of colonies when treated with tryptophan less than 10 mM was not significantly different from that of the control group, but the number of colonies when treated with 10 mM tryptophan was reduced to 80% of the control group. As in the case of P. acnes, tryptophan that is not photoactivated seems to inhibit the growth of S. aureus if its concentration is high enough (FIG. 2).

Example 2 Effect of Acne Improvement by Tryptophan and Light

On one face, the formulation containing 10% of tryptophan was applied, and the other face was irradiated with IPJ at 20J while no drug was applied. The study was repeated three times at two week intervals, and the number of inflammatory lesions was counted for improvement. Compared with the number of inflammatory lesions before treatment, the IPL-only group showed no statistically significant difference, but the tryptophan-treated and IPL-treated group showed a significant statistical difference. It has been shown to be effective in treatment (FIG. 3).

Example 3) Tryptophan and Light Sebaceous glands  Reduction effect

As in Example 2, the formulation containing 10% tryptophan was applied to one face, and the other face was irradiated with IPJ at 20 J without applying any drug. The study was repeated three times at the interval of two weeks, and sebum secretion was measured to observe the degree of improvement. When comparing sebum secretion levels with pretreatment, the IPL alone group showed no statistically significant difference in sebum secretion after 4 and 6 weeks of treatment, but the group treated with tryptophan and IPL showed significant statistical difference. , Tryptophan application and IPL irradiation showed an effect on sebum secretion inhibition (FIG. 4).

Example 4 Skin Stimulation Test

The skin irritation test was conducted by a paper published by Genji Imokawa et al. (Genji Imokawa and Yutaka Mishma, Nichihikaishi, 86 (8): 473, 1976), and closed using a Finn Chamber of Norgesplaster A / S Closed patch test was performed.

10 μl of the formulation containing 10% of tryptophan was absorbed into a paper disc fitted to a Finn Chamber and placed in a Finn Chamber for 48 hours on 10 adult male skins. After 48 hours, the Finn Chamber was immediately removed, washed with running tap water and dried, and the skin condition was determined after 2 hours.

In order to evaluate the degree of skin irritation according to light irradiation, as shown in Example 2, an agent containing 10% tryptophan was applied to one face, and the other side was irradiated with IPJ at 20J for 2 hours without applying any drugs. The condition of the skin was then determined.

Skin irritation was determined according to the following criteria:

Criteria (stimulation intensity)

           -: No change with the naked eye

           O: light redness

           +: Moderate redness

          ++: strong redness and edema

As a result, as shown in Table 1, the transdermal formulation of the present invention was shown to be not toxic at all by not irritating the skin.

Skin irritation judgment result 피부 Skin irritation evaluation by chamber method - Skin irritation evaluation during IPL irradiation after tryptophan application - Skin irritation evaluation after IPL irradiation without tryptophan -

Claims (9)

Comprising tryptophan or a pharmaceutically acceptable salt thereof,
Photo sensitizer for acne.
The method of claim 1,
The tryptophan exhibits photosensitizing activity against light rays ranging from 280 nm to 1000 nm,
Photo sensitizer for acne.
The method of claim 1,
The tryptophan is photoactivated in vitro or in vivo by ultraviolet light of 350 to 400 nm, or blue light of 400 to 500 nm, or green light of 500 to 600 nm,
Photo sensitizer for acne.
4. The method according to any one of claims 1 to 3,
Containing 0.001% to 30% by weight of tryptophan,
Photo sensitizer for acne.
Comprising tryptophan or a pharmaceutically acceptable salt thereof,
Photosensitive sebum secretion inhibitors.
6. The method of claim 5,
The tryptophan exhibits photosensitizing activity against light rays ranging from 280 nm to 1000 nm,
Photosensitive sebum secretion inhibitors.
6. The method of claim 5,
The tryptophan is photoactivated in vitro or in vivo by ultraviolet light of 350 to 400 nm, or blue light of 400 to 500 nm, or green light of 500 to 600 nm,
Photosensitive sebum secretion inhibitors.
8. The method according to any one of claims 5 to 7,
Containing 0.001% to 30% by weight of tryptophan,
Photosensitive sebum secretion inhibitors.
i) a composition containing tryptophan or a pharmaceutically acceptable salt having photosensitive activity in an amount of 0.001% to 30% by weight; And
ii) a kit for treating photodynamic acne and inhibiting photosensitive sebum secretion, comprising a light source generating a light ray having a wavelength ranging from 280 nm to 1000 nm.

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