JP2010229101A - Composition for skin - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an agent for preventing or treating itching skin diseases, which inhibits itches accompanied by itching skin diseases, such as atopic dermatitis, hives, contact dermatitis, psoriasis vulgaris, xeroderma, and prurigo, especially an agent for preventing or treating the atopic dermatitis, and is safe. <P>SOLUTION: It has been found that lactoferrin inhibits peripheral sensory nerve elongation caused by making a nerve growth factor to act on PC12 cells. The lactoferrin exhibits an effect for treating and preventing the pruritus of NC mouse which is a model animal of the atopic dermatitis. It is also found that an effect for clearly improving the skin symptoms of a patient in a therapeutic experiment performed under the agreement of the patient suffering from the atopic dermatitis has been shown. Namely, the composition for the skin safely and quickly inhibits itches accompanied by itching skin diseases such as intractable atopic dermatitis, hives, contact dermatitis, psoriasis vulgaris, xeroderma, and prurigo. Thereby, the composition for the skin, containing the lactoferrin as an active ingredient can safely treat and prevent the skin diseases accompanied by an intractable itching sense, including the atopic dermatitis. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a prophylactic or therapeutic agent for pruritic skin disease containing lactoferrin, particularly a prophylactic or therapeutic agent for atopic dermatitis.

  Pruritic skin diseases include atopic dermatitis, urticaria, contact dermatitis, psoriasis vulgaris, psoriasis and urticaria, but are a diverse group of diseases with different etiologies. Histamine has long been known as a stagnation-inducing substance, and antihistamines have been used to suppress itching. However, since there are many itches in which antihistamines are ineffective, it is clear that there is a itch mechanism that does not involve histamine. Usually, in normal skin, the peripheral sensory nerves that convey itch reach only the basal layer, the boundary between the epidermis and dermis, but psoriasis, atopic dermatitis, psoriasis vulgaris, allergic contact dermatitis Have been reported that a large number of peripheral sensory nerves that convey itch have extended into the epidermis, which contributes to severe itch (see Non-Patent Documents 1 to 5). It is considered that nerve growth factor (hereinafter abbreviated as NGF) produced by keratinocyte cells is mainly involved in the peripheral sensory nerve elongation (see Non-Patent Document 6). PC12 cells are passage cells derived from rat adrenal medullary pheochromocytoma. Since these cells differentiate into nerve-like cells upon stimulation with NGF, they are frequently used for evaluating the action on nerve cells (see Non-Patent Document 7).

  Lactoferrin is a glycoprotein with a molecular weight of about 80,000 daltons isolated as a red protein from milk in 1939. Lactoferrin is abundant in milk, especially colostrum, and human newborns ingest 5-7 grams per day. In addition to being contained in mature neutrophils as lactoferrin granules, it is also contained in exocrine fluids such as tears, saliva, pancreatic juice and vaginal secretions. One of the inventors of the present invention has found that lactoferrin enhances the action of endogenous opioids when pursuing a mechanism of action because of its central and peripheral analgesic effect (Non-patent Document 8). ). Orally administered lactoferrin crosses the blood-brain barrier and is taken into the cerebrospinal fluid, and thus has been shown to have an analgesic effect on stubborn migraine (Non-Patent Document 9). Furthermore, lactoferrin is a type 1 produced when a mononuclear cell is stimulated with an oligonucleotide containing (5 ′) cytosine, phosphate, and guanine (3 ′) when taken orally for 1 to 4 weeks as a liposome preparation. Since interferon is significantly increased, an effect of suppressing viral infection, for example, influenza infection is expected (Non-patent Document 10). In addition, lactoferrin is known to have an improving effect on allergic rhinitis (Non-patent Document 11) and rheumatoid arthritis (Non-patent Document 12) when administered orally. It is a multifunctional protein that is known to have an effect on whitening and rough skin (see Patent Document 1). As an effect of lactoferrin on pruritic skin diseases including atopic dermatitis, only lactoferrin is described as one of a huge group of compounds (Patent Document 2), and the effect, safety, dosage of administration, etc. Specificity was lacking.

  The lactoferrin of this patent is not limited to bovine lactoferrin extracted from fresh milk, lactoferrin extracted from milk of sheep, goats, horses, camels, etc., lactoferrin produced by a recombinant microorganism of human lactoferrin gene and transgenic mammals Includes lactoferrin produced by

JP-A-11-302150 Japanese Patent Publication No. 9-501925

Tobin D. et al. J. Allergy Clin. Immunol. 1992; 90: 613-622 Urashima R. et al. Virchows Arch. 1998; 432: 363-370 Masahiko Toyoda et al., JSHI Journal 1997; 107: 1257-1279 Kenji Takamori, Shinkin 2000; 54: 52-56 Kinkelin I. et al. Cell Tissue Res. 2000; 302: 31-37 Pincelli C. et al. J Invest Dermatol Symp Proc. 2: 31-36 Greene LA. Et al. Proc Natl Acad SciUSA. 1976; 73, 2424-2428. Hayashida K. et al. Am J Physiol Regul Integr Comp Physiol. 2003; 285 (2): R306-12 Ji B et al. Life Sci. 2006; 78: 851-55. Ishikado A. et al. Biofactors. 2004; 21: 69-72. Kruzel ML. Et al. Immunology. 2006; 119: 159-66. Hayashida K. et al. J Vet Med Sci. 2004; 66: 149-54

  The present invention provides a preventive or therapeutic agent for pruritic skin disease that suppresses pruritus of pruritic skin disease and has no side effects, particularly a prophylactic or therapeutic agent for intractable skin diseases such as atopic dermatitis and psoriasis vulgaris The purpose is to do.

  As a result of intensive studies to solve the above problems, the present inventors have found that lactoferrin suppresses peripheral sensory nerve elongation by nerve growth factor (NGF) and has an excellent improvement effect on pruritic skin diseases. The present invention has been completed.

  PC12 cells are frequently used to evaluate the effects on nerve cells. Lactoferrin suppresses NGF-induced differentiation of PC12 cells into peripheral sensory nerves and nerve fiber elongation. In other words, lactoferrin is percutaneously absorbed into the epidermis, suggesting the possibility of suppressing the extension of peripheral sensory nerve fibers that cause severe itching into the epidermis.

  Furthermore, the present inventors used NC / Nga mice that spontaneously develop atopic dermatitis-like skin lesions, one of pruritic skin diseases, by continuously administering lactoferrin by the percutaneous route and the oral route. Then, the preventive and therapeutic effects on cervical dorsal dermatitis considered to be caused by scratching behavior of hind limbs were evaluated. As a result, it was found that lactoferrin has a preventive and therapeutic effect on NC / Nga mouse dermatitis. Furthermore, even in a clinical trial conducted with the consent of atopic dermatitis patients, when macrogol ointment containing 0.15-0.5% lactoferrin was applied to the affected area, itching and redness after 3 to 7 days In addition, symptoms of atopic dermatitis such as eczema were improved.

  Atopic dermatitis becomes chronic and many patients are bothered. Adrenocortical steroid hormones commonly used for the treatment of atopic dermatitis, when administered orally or transdermally, improve the pathological condition dramatically, although temporarily. However, because the treatment is prolonged and treatment is prolonged, steroids are worsened by immunosuppression, hirsutism, skin atrophy, redness, depigmentation, dilation of capillaries, steroid acne, mouth corner Various side effects such as inflammation, eczema / keratosis, worsening warts and purpura occur and cannot be used for a long time. The most important side effect of corticosteroids is that they cannot play their original role as stress buffers. That is, corticosteroid hormones are a symptomatic treatment that surpasses temporarily, and there is a need for a therapeutic agent that can be used safely and for a long period of time and that reliably improves skin symptoms. The present invention meets medical unmet needs as a preventive or therapeutic agent for pruritic skin diseases containing lactoferrin and a prophylactic or therapeutic agent for atopic dermatitis containing lactoferrin.

  According to the present invention, extension of peripheral sensory nerve fibers to the epidermis can be suppressed, and pruritic skin diseases can be prevented or treated, and in particular, atopic dermatitis can be prevented and treated for a long period of time.

  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, cosmetics and the like for the purpose of preventing pruritus caused by pruritic skin disease.

  The lactoferrin in the present invention is used as an oral preparation, an external preparation, a suppository, a food additive, and a cosmetic. Lactoferrin is rapidly hydrolyzed when pepsin is present in the stomach at a pH of 3.5 or lower. Therefore, when administered as an oral preparation, enteric preparations that disintegrate in the small intestine through the stomach and release lactoferrin, or internal use A sustained-release preparation that gradually releases lactoferrin from about 2 hours later is desirable. For topical application, ointments, creams, lotions, gauze adhesive bandages (eg “BandAid” from Johnson & Johnson), emulsions (emulsions, suspensions, etc.), foams, blends Phase or amphiphilic emulsion system (oil / water-water / oil-mixed phase) cream, liposome, paste, powder, etc. The present invention can be applied to nasal mucosal administration using suppositories and powders, rectal administration using suppositories, and the like.

  The effective dose of lactoferrin can be appropriately increased or decreased depending on the patient's body weight, age, sex, etc. In the case of oral administration, it is 0.01-10 g as lactoferrin per day, and preferably 0.1-1 g. It is administered once to several times a day. The concentration range when applied to the skin as a dermatological composition is 0.0001% to 5%, preferably 0.001% to 2%.

  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, baths, 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. It is lactoferrin.

  5 g of lactoferrin, 5 g of lipid obtained from milk fat globule membrane, 50 g of liquid paraffin (# 70), 100 g of squalane, 60 g of cetostearyl alcohol, 20 g of beeswax, 15 g of glyceryl monostearate, 20 g of sorbitan monolaurate, 2 g of methylparaben, 1 g of propylparaben, Appropriate amount of purified water. The above ingredients were mixed and uniformly emulsified, and an appropriate amount of fragrance was added to obtain 500 g of a cream.

  1 g of lactoferrin, 5 g of 1,3-butylene glycol, 2 g of lactic acid, 50 g of ethanol, 3 g of benzalkonium chloride, and appropriate amounts of purified water were mixed together to obtain 500 g of a lotion preparation.

  Lactoferrin 3000g, lactose 1870g, microcrystalline cellulose 1000g, talc 50g, sucrose fatty acid ester 50g, magnesium stearate 30g and weighed and mixed uniformly. Tablets were obtained by tableting to 300 mg. The tablets were sprayed with water-soluble shellac (Freund Sangyo) using a high coater granulator (Freund Sangyo) and coated with an enteric coating to obtain lactoferrin enteric tablets.

  Add 1 g of lactoferrin spray-dried powder (DMV, purity 90%, average particle size 5 μm) to 99 g of macrogol ointment base (1: 1 mixture of macrogol 400 and macrogol 4000) and mix thoroughly in a mortar. To make macrogol ointment.

Test Example 1: Inhibitory effect on NGF-induced PC12 cell process elongation (test method) 100 mg of lactoferrin was dissolved in 100 ml of PBS to prepare 1 mg / ml aqueous solution as lactoferrin. PC-12 cells were cultured in DMEM medium at 37 ° C. and 5% CO2. 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 containing differentiation medium (serum-free medium containing 300 ng / mL 7S-NGF and / or 10 μg / mL lactoferrin), and 100 μL of cell suspension was added to each chamber. After culturing for 3 days, the extension of the process was stained with a neurite stain, the cells at the top of the chamber were wiped off, eluted with a neurite eluate, and the absorbance at 540 nm was measured.

  (Test result) NGF added group value is 0% inhibition rate, NGF non-added (control) PC-12 cells do not extend dendrites as nerve cells, so when the value is 100%, it becomes NGF The inhibition rate in the medium supplemented with 10 μg / mL of lactoferrin was as high as 95%. From this result, it was found that lactoferrin significantly suppressed NGF-induced neuronal cell growth.

Test Example 2: Therapeutic effect on NC / Nga mouse atopic dermatitis-like symptoms (test method) A lactoferrin ointment prepared by the same method as in Example 4 was used in the test. In the test, NC / Nga male mice that developed dermatitis having a body weight of about 30 g and 8 per group were used as test animals in both the control group and the lactoferrin group. In the lactoferrin group, 0.1 g of lactoferrin ointment was applied to the back of the neck once a day for 4 weeks, and only macrogol ointment containing no lactoferrin was applied to the control group. Observation of skin symptoms in each group was performed once / week. Skin symptoms were recorded according to the criteria shown in Table 1, and a significant difference test was performed. The number of spontaneous scratching behaviors was also measured in the same manner (FIG. 2). Statistical analysis of judgment and measurement results was performed by Student's unpaired t-test, and the significance level was 5%.

  The skin symptoms of the face and auricle were scored according to the evaluation criteria shown in Table 1, and the numerical value of each group was expressed as an average value ± standard error. The significant difference was tested by Student's unpaired t-test.

  (Test result) The lactoferrin ointment application group significantly suppressed dermatitis of NC / Nga mice, and the suppression rate of the dermatitis score at 4 weeks was 94% (P <0.01). The number of scratching behavior, which is an index of itchiness, was also significantly suppressed, and the suppression rate of scratching behavior 28 days after the start of the experiment was 58% (P <0.05). Thus, lactoferrin ointment was found to improve the symptoms of atopic dermatitis.

  (Clinical trial 1) With the consent of a patient suffering from refractory atopic dermatitis for many years (a female office worker 38 years old), a macrogol ointment containing 0.15% lactoferrin was applied. I instructed patients who exchanged consent forms like molds to apply 10 g of lactoferrin ointment every week, and apply ointment to the affected area three times daily. The patient was interviewed and photographs of the affected area were taken once a week, and the progress was observed over 12 weeks.

(Results) Before treatment, atopic dermatitis characterized by skin redness and inflammation surrounding the mouth was observed. In addition, there was stomatitis in the right corner of the mouth, and the lips were rough, resulting in a cold sore infection. By applying a macrogol ointment containing 0.15% lactoferrin, these symptoms were greatly improved when visited 6 weeks later.

  (Clinical Trial 2) A woman (35 years old) who developed hay fever before school age took 2 tablets of enteric lactoferrin tablets prepared in Example 3 three times a day from March 2003. Before she took an enteric lactoferrin tablet, she was serious, taking antihistamines every day during the hay fever season, and wearing goggles and a mask to protect her eyes and nose from pollen when going out. Taking antihistamines improved redness of the eyes, nasal congestion, and a large amount of runny nose, but no general itching. This is a case where antihistamines did not improve itching. From the year of taking the enteric lactoferrin tablet, the redness of the eyes, nasal congestion, and runny nose were settled, and at the same time it was not possible to feel itching. Usually, it is said that hay fever is not cured, but in the 2007 pollen season, even if enteric lactoferrin tablets were not taken, symptom of hay fever disappeared and it was diagnosed that hay fever was cured.

  The prophylactic or therapeutic agent for pruritic skin disease of the present invention can be used as a very useful prophylactic or therapeutic agent for pruritic skin disease based on an unprecedented concept of systemic or local administration of lactoferrin.

Claims (4)

A prophylactic or therapeutic agent for pruritic skin diseases containing lactoferrin. 2. The agent for preventing and treating pruritic skin disease according to claim 1, wherein the pruritic skin disease is atopic dermatitis. The prophylactic / therapeutic agent for pruritic skin disease according to claim 1 or 2, wherein the prophylactic / therapeutic agent for pruritic skin disease is an external preparation. The prophylactic / therapeutic agent for pruritic skin disease according to claim 1 or 2, wherein the prophylactic / therapeutic agent for pruritic skin disease is a bath agent.