JP5256578B2 - Preventive or therapeutic agent for pruritic skin disease - Google Patents

Description

  The present invention relates to a prophylactic or therapeutic agent for pruritic skin disease, particularly a prophylactic or therapeutic agent for atopic dermatitis, which contains gafshokuso.

  Pruritic skin diseases are various diseases accompanied by itch such as atopic dermatitis, urticaria, contact dermatitis, xerosis, and urticaria. Histamine has long been known as a stagnation-inducing substance, and antihistamines have been used to suppress itching. However, since there are many reports of itching that antihistamines are ineffective, it is clear that there is a mechanism of itching that does not involve histamine, and research is currently underway from various viewpoints. Normally, in normal skin, the peripheral sensory nerves that convey itch extend only to the border between the epidermis and dermis, but in dry skin, atopic dermatitis, psoriasis, allergic contact dermatitis, etc. In recent years, it has been reported that the peripheral sensory nerves that are transmitted have extended into the epidermis, and this contributes to severe itching (see Non-Patent Documents 1 to 5). It is considered that nerve growth factor (hereinafter referred to as NGF as appropriate) produced mainly from keratinocyte cells is involved in the peripheral sensory nerve elongation (see Non-Patent Document 6). PC12 cells derived from rat adrenal medullary pheochromocytoma are frequently used for evaluating the action on nerve cells because PC12 cells differentiate into nerve-like cells upon stimulation with NGF (see Non-patent Document 7).

  Gafushokusou (鵞 鵞 不 食草) is a dried whole plant of the Asteraceae kingfisher (aka Centipeda minima). Gafushoku is a herbal medicine that has long been known to be effective against headaches, colds, allergic rhinitis and rheumatoid arthritis. Moreover, the effect | action with respect to a whitening effect and rough skin is known (refer patent document 1). However, the effect on pruritic skin diseases such as atopic dermatitis is not known.

JP 2000-247829 A Tobin, D., Nabarro, G., de la Faille, HB, van Vloten, WA, van der Putte, SCJ, Schuurman, H.-J., 1992. Increased number of immunoreactive nerve fibers in atopic dermatitis. J. Allergy Clin. Immunol. 90, 613-622. Urashima, R., Mihara, M., 1998. Cutaneous nerves in atopic dermatitis. Virchows Arch. 432, 363-370 Masahiko Toyoda et al., 1997, Changes in the cutaneous nervous system with cyclosporine administration in patients with atopic dermatitis, JSJ 107 (10): 1257-1279 Kenji Takamori, 2000, mechanism of itchiness by dry skin, skin 54 (5 increase): 52-56 Kinkelin, I., Motzing, S., Koltenzenburg, M., Brocker, E.B., 2000. Increase in NGF content and nerve fiber sprouting in human allergic contact eczema.Cell Tissue Res. 302, 31-37 Pincelli, C., Yaar, M., 1997. Nerve growth factor: Its significance in cutaneous biology.J Invest Dermatol Symp Proc. 2, 31-36 Greene, L.A., Tischler, A.M., 1976.Establishment of a noradrenergic clonal line of rat adrenal pheochromocytome cells which respond to nerve growth factor.Proc Natl Acad Sci USA.73, 2424-2428.

  An object of the present invention is to provide a preventive or therapeutic agent for pruritic skin disease, particularly an atopic dermatitis preventive or therapeutic agent, which suppresses itching associated with pruritic skin disease.

  As a result of intensive studies to solve the above problems, the present inventors have suppressed peripheral sensory nerve elongation caused by NGF, and atopic dermatitis, urticaria, contact dermatitis, psoriasis, urticaria, etc. Was found to have an excellent ameliorating effect on pruritic skin diseases.

  As a result of evaluating the inhibitory effect on peripheral sensory nerve elongation induced by NGF, the present inventors used PC12 cells that are frequently used for evaluating the action on nerve cells because they have properties very similar to those of nerve cells, It was found that Gafushokuso strongly inhibits peripheral sensory nerve elongation caused by NGF.

  Furthermore, the present inventors used a NC / Nga mouse that spontaneously develops atopic dermatitis-like skin lesions, which is one of pruritic skin diseases, by continuously administering gafushoxu, We evaluated the preventive and therapeutic effects on cervical dorsal dermatitis, which may be caused by As a result, it was found that Gafushokuso has a very strong NC / Nga mouse dermatitis prevention and treatment effect. It was also found that spontaneous scratching behavior, which is an index of itchiness, and transdermal moisture transpiration, which is an index of skin barrier function, have similarly strong preventive and therapeutic effects.

That is, the present invention
1. It is a prophylactic or therapeutic agent for pruritic skin diseases containing gafshokuso.
2. It is a preventive or therapeutic agent for atopic dermatitis containing gafushoxou.

  According to the present invention, it is possible to suppress peripheral sensory nerve elongation by NGF and prevent or treat pruritic skin diseases, and in particular, prevent and treat atopic dermatitis.

  The agent for preventing or treating pruritic skin disease as used in the present invention is used as a pharmaceutical, food for specified health use, food, etc. for the purpose of preventing pruritus caused by pruritic skin disease.

  In the present invention, gafushoku is used in the form of a herbal powder and a herbal extract. A herbal extract can be obtained using water, ethanol, propylene glycol, 1,3-butylene glycol or a mixture thereof as an extraction solvent.

  The effective dose of gafushox can be appropriately increased or decreased depending on the patient's weight, age, sex, etc., but is 0.01 to 10 g, preferably 0.1 to 1 g, per day as the amount of drug substance. Can be administered one to several times a day.

  The present invention contains additives such as excipients, disintegrants, binders, lubricants, antioxidants, coating agents, colorants, flavoring agents, surfactants, plasticizers, etc. Can be made into granules, powders, capsules, tablets, chewable tablets, dry syrups, solutions, ointments, creams, patches.

  In addition, an antihistamine, an antiallergic agent, a local anesthetic, an anti-inflammatory agent, a steroid agent, a moisturizer, a bactericidal agent, a refreshing agent, vitamins, and other herbal medicines can be added as long as the effects of the present invention are not impaired. .

  Hereinafter, the present invention will be described in more detail with reference to examples and test examples. In addition, Gafushokuso is blended as an extract, and the blending amount is the amount of the drug substance.

Gafushoku 50g
5g sodium edetate
Liquid paraffin (# 70) 50g
Squalane 100g
Setostearyl alcohol 60g
20g of beeswax
15g glyceryl monostearate
20g sorbitan monolaurate
Methyl paraben 2g
1g propylparaben
Purified water appropriate amount Each of the above components was mixed and uniformly emulsified, and a proper amount of perfume was added to obtain 500 g of a cream.

Gafushoku 10g
1,3-butylene glycol 5g
Lactic acid 2g
50g ethanol
Benzalkonium chloride 3g
Purified water appropriate amount The above ingredients were mixed and homogenized to obtain 500 g of a lotion preparation.

Gafushoku 30g
Lactose 1870g
500g microcrystalline cellulose
Low substituted hydroxypropylcellulose 500g
Talc 50g
Hardened castor oil 50g
Each of the above components and amounts were weighed and mixed uniformly, and then the obtained mixed powder was tableted to give a tablet weight of 300 mg by a direct compression method to obtain a tablet.

Gafushoku 25g
Lactose 425g
450g microcrystalline cellulose
Talc 50g
After weighing and mixing the above components and amounts uniformly, the obtained mixed powder was filled into No. 1 hard capsules in an amount of 250 mg to obtain capsules.

Test Example 1: Inhibitory effect on NGF-induced PC12 cell process elongation (test method)
After chopping gafsoxo, 10 times the amount of 50% ethanol was added, and the mixture was extracted by heating at about 80 ° C. After filtration, the ethanol was distilled off under reduced pressure, and further concentration was carried out to obtain a gafusoxou extract. PC-12 cells were cultured in DMEM medium containing 10% FBS, 5% HS, 50 U / mL penicillin and 50 μg / mL streptomycin at 37 ° C. under 5% CO ₂ . The protrusion elongation was quantified using the Neuroite Outgrowth Quantification Assay Kit (CHEMICON, NS200). After adding type I collagen solution to the plate, the chamber was immersed and incubated at 37 ° C. for 2 hours. During this time, the cells were peeled off with a versene solution, and the viable cells were adjusted to a concentration of 2 × 10 ⁶ cells / mL. The chamber was transferred to a plate supplemented with differentiation medium (serum-free medium containing 300 ng / mL 7S-NGF and / or 100 μg / mL gaffoxo extract), and 100 μL of cell suspension was added to each chamber. After culturing for 3 days, the neurite outgrowth was stained with neurite staining solution, the cells at the top of the chamber were wiped off, eluted with neurite eluate, and the absorbency at 540 nm was measured and quantified.

(Test results)
When the value of the NGF added group was 0% and the value of the non-added group (control) was 100%, the inhibition rate of the Gafshox extract was as high as 100.2%. From these results, it was suggested that gafushokoku extract strongly suppresses the nerve cell elongation induced by NGF.

Test Example 2: Preventive action against NC / Nga mouse atopic dermatitis-like symptoms (test method)
A test gaffoxo extract prepared in the same manner as in Test Example 1 was used for the test. A 50% ethanol was used as a solvent, and this extract was used as a test gaffoxo extract prepared to a drug substance amount of 10% (w / v). As a reagent, 50% ethanol as a control drug and a test gaffoxo extract were used. As test animals, NC / Nga male mice having a body weight of about 30 g and having no dermatitis, 8 mice per group were used. Mice were allowed to live with animals with dermatitis for a week to induce atopic dermatitis-like symptoms. Thereafter, 100 μl of 50% ethanol was applied to the control group, and 100 μl of the test gaffoxo extract was applied to the back of the neck once a day for 7 weeks (5 days after continuous application, 2 days without application). The skin symptoms in each group were observed before each weekly application and at the end of the application test. The rostral dorsal skin symptom was classified into 7 levels according to the criteria shown in Table 1, and determined as 0 to 6 points (FIG. 1). In addition, the number of spontaneous scratching behaviors was measured for 24 hours after the end of continuous application using a pruritus behavior measurement system (NS-SCT16, Neuroscience) (FIG. 2). Furthermore, the transdermal water transpiration rate at the back of the neck was measured using a tevameter (TM210, Integral) for 24 hours after the last observation of skin symptoms (FIG. 3). Statistical analysis of judgment and measurement results was performed by Student's t-test, and the significance level was 5%.

(Test results)
The test gafushoxou extract administration group significantly suppressed the onset of dermatitis in NC / Nga mice. In addition, the number of scratching behavior, which is an index of itchiness, and the amount of transdermal moisture transpiration, which is an index of skin barrier function, were significantly suppressed. Therefore, it became clear that Gafushoku prevents the onset of atopic dermatitis. (*: P <0.05, **: P <0.01)

Test Example 3: Treatment for NC / Nga mouse atopic dermatitis-like symptoms (test method)
A test gaffoxo extract prepared in the same manner as in Test Example 1 was used for the test. In the test, NC / Nga male mice having a body weight of about 30 g and 8 mice per group were used as test animals in both the control group and the gaffox group. In the gaffoxo group, 100 μl of test gaffoxo extract was applied to the back of the neck once a day for 4 weeks (5 days after continuous application, 2 days without application). Observation of skin symptoms in each group was performed once / week. The skin symptoms on the rostral dorsal region were determined according to the same criteria as in Test Example 1 (FIG. 4). Moreover, the number of spontaneous scratching behaviors was measured in the same manner as in Test Example 1 (FIG. 5). Statistical analysis of judgment and measurement results was performed by Student's t-test, and the significance level was 5%.

(Test results)
The test gafushoxo extract administration group significantly suppressed dermatitis in the onset NC / Nga mice. Furthermore, the number of scratching behavior, which is an index of itchiness, was also significantly suppressed. Therefore, it has been clarified that Gafushoxou extract treats the symptoms of atopic dermatitis. (*: P <0.05, **: P <0.01)

Test Example 4: NC / Nga mouse treatment for atopic dermatitis-like symptoms (oral administration)
(Test method)
A test gaffoxo extract prepared in the same manner as in Test Example 1 was used for the test. A test gaffo extract was prepared using 0.2% carboxymethylcellulose so as to give a dose of 1000 mg / kg, and used for the test. As test animals, NC / Nga male mice having a body weight of about 30 g and 8 mice per group were used. The test Gafshoxo extract was orally administered once a day for 4 weeks (5 days after continuous administration, 2 days after non-administration). The observation of skin symptoms in each group was performed once a week 5 days after continuous administration of each reagent.

  A total of three skin symptoms of the face, auricle and rostral back were classified into four levels according to the criteria shown in Table 2, and each scored 0 to 3 points, and the total was used as an index (FIG. 6). Statistical analysis of the judgment results was performed by Student's t-test, and the significance level was 5%.

(Test results)
The test gafushoxo extract administration group significantly suppressed dermatitis in the onset NC / Nga mice. Therefore, it has been clarified that gafushoxou extract treats the symptoms of atopic dermatitis even by internal use. (*: P <0.05, **: P <0.01)

  The preventive or therapeutic agent for pruritic skin disease of the present invention can be used as a very useful preventive or therapeutic agent for pruritic skin disease, based on an unprecedented concept of returning normal excess sensory nerves to normal.

It is the result of having observed the dermatitis onset inhibitory effect of a NC / Nga mouse, the dermatitis score was shown on the vertical axis, and the test period was shown on the horizontal axis. It is the result of measuring the scratching behavior inhibitory effect of NC / Nga mice, and the vertical axis represents the number of scratching behaviors and the horizontal axis represents the administration group. It is the result of measuring the transdermal water transpiration suppression effect of NC / Nga mice, where the vertical axis represents the transdermal water transpiration and the horizontal axis represents the administration group. It is the result of having observed the dermatitis inhibitory effect of the onset NC / Nga mouse, The dermatitis score was shown on the vertical axis | shaft and the test period was shown on the horizontal axis. It is the result of measuring the scratching behavior inhibitory effect of the onset NC / Nga mouse, and the vertical axis represents the number of scratching behavior and the horizontal axis represents the administration group. It is the result of measuring the scratching behavior inhibitory effect of NC / Nga mice that developed by oral administration, the number of scratching behaviors on the vertical axis, and the administration group on the horizontal axis.

Claims (1)

A prophylactic or therapeutic agent for atopic dermatitis or xeroderma based on a peripheral sensory nerve elongation-inhibiting action, characterized by containing gafushoxo.

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