JPH08325155A - Immunosuppression improver - Google Patents

Abstract

PURPOSE: To obtain an immunosuppression improver that contains aloesin having excellent ameliorative effect against photoimmunosuppression. CONSTITUTION: This immunosuppression improver contains aloesin, a hydroxy- chromone glycoside which is obtained from Aloe by separation and purification. Aloesin is obtained, for example, by squeezing the above-ground part of Aloe with a juicer mixer, centrifuging the squeeze, adding an equal amount of alcohol to the squeeze to precipitate and remove the polysaccharide, concentrating the filtrate to, dryness, dissolving the concentrate in water, subjecting the solution to the partition extraction with n-butanol and purifying the fraction with a silica gel column. This improver is made into a preparation for external use and the dose of aloesin is 20-500mg/day, preferably 50-200mg/day in 5-6 portions. The acute toxicity of the preparation, LD50, is over 250mg/kg based on aloesin.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an agent for improving immunosuppression, which contains aloesin.

[0002]

2. Description of the Related Art Chronic UV exposure to the skin reduces the general and local immunity of the skin and induces photoimmunosuppression. Further, it is known that photoimmunosuppression is one of the causes of skin cancer occurrence and infectious diseases, and it is considered important to use an immunosuppression-improving agent in order to improve the deterioration of immune function of the skin. . Aloesin is used as a tyrosinase inhibitor (Japanese Patent Laid-Open No. 62-286909), but there is no application example as an immunosuppressant improving agent.

[0003]

Then, when the effectiveness of aloesin was examined, the present invention was completed because it showed an excellent improving effect on photoimmunosuppression.

[0004]

The present invention is an immunosuppression-improving agent characterized by containing aloesin. Aloesin is a hydroxychromone glycoside and can be separated and purified from aloe by the method of Conner et al. (Phytochemistory 29 (3), 941-944, (1990)).

For example, the above-ground portion of aloe is squeezed using a juicer mixer or the like, and then a juice is obtained by a treatment such as centrifugation. Subsequently, an equal amount of alcohol (ethanol, methanol, etc.) is added to precipitate the polysaccharide, and the polysaccharide is removed by centrifugation or filtration, and then concentrated to dryness. An equal amount of n-butanol is added to the product dissolved in water, and partition extraction is performed to obtain a butanol fraction. This butanol fraction can be applied to a silica gel column, a Sephadex LH-20 column, an ODS column or the like to obtain aloesin. The aloesin in the present invention does not necessarily have to be completely purified, and may be a crudely purified product. Further, in order to improve its stability, it is possible to add an antioxidant or the like.

According to the present invention, oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, ultraviolet rays used in cosmetics and medicines are used as long as the effects of aloesin are not impaired. Raw materials such as an absorbent can be blended. Further, the dosage form of the immunosuppressive improving agent of the present invention is an external preparation, a lotion, a water-soluble ointment, an oily ointment,
Emulsive ointment and the like can be mentioned, and it is produced according to a usual formulation technique. The daily dose is 20 mg as aloesin.
~ 500 mg, preferably 50 mg-200 mg, can be administered in 5-6 divided doses. The LD50 for acute toxicity was 250 mg / kg or more as aloesin.

[0007]

EXAMPLES Next, examples will be given to explain the present invention in detail, but the present invention is not limited thereto. The parts of the compounding amounts shown in the examples are parts by weight. Aloesin is
It was prepared by the following method. (Production Example 1) Aloe extract was obtained by squeezing the above-ground portion of 20 kg of Aloe ferox Miller and continuously centrifuging at 10,000 rpm. Subsequently, twice the amount of ethanol was added to the supernatant, the precipitate was removed by centrifugation, and the mixture was concentrated to dryness. After this was dissolved in water, an equal amount of n-butanol was added to carry out partition extraction. This operation was repeated 3 times and the butanol layers were collected to obtain a butanol fraction. The obtained fractions were concentrated to dryness, and then subjected to preparative high performance liquid chromatography using an ODS column (solvent: 30% aqueous methanol solution, flow rate: 40 ml / min, detection: 295 nm) to separate 1 g of aloesin. The obtained aloesin was identified by measuring its NMR spectrum and comparing it with literature values.

Example-1 Lotion formulation Formulation amount 1. Stearyl alcohol 2.5 2. Liquid paraffin 25.0 3. Sodium lauryl sulfate 1.0 4. Propylene glycol 12.0 5. Methyl paraoxybenzoate 0.025 6. Propyl paraoxybenzoate 0.025 7. Aloesin 2.0 8. Purified water 57.45 Manufacturing method: 70 parts of oil phase components 1-2 and water phase components 3-8, respectively.
After melting by heating to ~ 75 ℃, water phase components 3-8 in oil phase components 1-2
Add to emulsify and cool to 30 ° C to obtain the product.

Example-2 Water-soluble ointment Formulation amount 1. Polyethylene glycol 4000 (PEG 4000) 50.0 2. Polyethylene glycol (PEG 400) 49.5 3. Aloesin 0.5 Production method: Dissolve each component uniformly to obtain a product.

Example-3 Oil and Ointment Formulation Amount 1. Purified lanolin 5.0 2. White beeswax 5.0 3. Aloesin 2.0 4. White petrolatum 88.0 Manufacturing method: Each component is uniformly dissolved to obtain a product.

Example-4 Emulsion Ointment-1 Prescription Compounding amount 1. White petrolatum 25.0 2. Stearyl alcohol 22.0 3. Propylene glycol 12.0 4. Sodium lauryl sulfate 1.5 5. Ethyl paraoxybenzoate 0.025 6. Propyl paraoxybenzoate 0.015 7. Aloesin 2.0 8. Purified water 37.46 Manufacturing method: 70 parts of oil phase components 1-2 and water phase components 3-8, respectively.
After melting by heating to ~ 75 ℃, water phase components 3-8 in oil phase components 1-2
Add to emulsify and cool to 30 ° C to obtain the product.

Example-5 Emulsion Ointment-2 Formulation Amount 1. Cholesterol 3.0 2. Stearyl alcohol 3.0 3. White beeswax 8.0 4. Aloesin 1.0 5. White petrolatum 85.0 Manufacturing method: 70-75 ° C. by mixing each component After melting by heating to
Cool to 30 ° C to obtain the product.

[0013]

INDUSTRIAL APPLICABILITY The agent for improving immunosuppression, which contains aloesin of the present invention, shows an excellent improving effect. Next, in order to explain the effect of the present invention in detail, an experimental example will be given. Experimental example Effect of aloesin on photoimmunosuppression induced by UV irradiation a) Test animal Specific-pathogen-free C57BL / 6Ncrj mouse (Japan SL
C) (male) 10-11 weeks old was used. The experimental group is n
= 6 b) Test sample The emulsion ointment-1 of Example-4 and the conventional emulsion ointment of Comparative example-1 (aloesin of Example-4 was replaced with purified water) were used. c) Ultraviolet irradiation Ultraviolet (UV-B) used Toshiba FL20S ・ E. For local immune suppression against contact hypersensitivity (CHS) reaction by UV-B irradiation, the back of the mouse is shaved and aluminum tape is applied to both ears.
Protected against inflammation by UV-B irradiation. After that, 4 to the mouse
UV-B of 400 J / m ² was continuously irradiated for a day. For systemic immune suppression, after shaving the mouse, protect both ears with aluminum tape and UV-B at 10 kJ / m ^2.
Was irradiated once. d) Induction of contact hypersensitivity reaction As an allergen, FITC (Fluorescein isothiocyanate) was dissolved in acetone: di-n-butyl phthalate (1: 1 v / v) at a concentration of 0.5% and used. For local immunosuppression experiments,
Six hours after UV-B irradiation on the 4th day, 0.5% FITC was placed on the back of the mouse.
50 μl was applied and sensitized. In the systemic immunosuppression experiment, the same amount as in the case of the local immunosuppression experiment was applied to the abdomen of the mouse 3 days after UV-B irradiation for sensitization. 5 days after sensitization, 10 μl of 0.5% FITC was applied to the front and back of each mouse's ear for both local and systemic challenge. Ear thickness was measured using a micrometer (Mitutoyo) before and 24 hours after the challenge.
The degree of ear edema was compared to unsensitized mice. e) Aloe application treatment 2 mg / cm ² of each sample was applied to the irradiated skin within 5 minutes after each UV-B irradiation. Margin below

[0014]

[Table 1] Effect of using aloesin obtained from "Production Example 1" on photoimmunosuppression (assuming 100 as control) ─────────────────── ────────────────── Test substance UV-B Local immunosuppression Systemic immunosuppression Improvement effect (%) Improvement effect (%) ─────────── ───────────────────────── None (control) -100.0 100.0 None + 0.0 0.0 Ointment (Comparative Example-1) + 20.2 25.0 Aloesin (Example) -4) ＋ 68.5 71.1 ──────────────────────────────────── As shown above, aloecin The immunosuppression-improving agent of the present invention containing the same showed an excellent improving effect on photoimmunosuppression.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display area A61K 35/78 A61K 35/78 V

Claims (1)

[Claims] 1. An agent for improving immunosuppression, which comprises aloesin.