WO1997035557A1

WO1997035557A1 - External skin-care composition

Info

Publication number: WO1997035557A1
Application number: PCT/JP1997/000880
Authority: WO
Inventors: Takatoshi Murase; Tadashi Hase; Yusuke Shibuya; Yoshinori Nishizawa; Ichiro Tokimitsu
Original assignee: Kao Corporation
Priority date: 1996-03-22
Filing date: 1997-03-18
Publication date: 1997-10-02

Abstract

An external skin-care composition for preventing and improving peeling, pachymenia, skin roughness, disorder of skin texture, pigmentation, degeneration or destruction of dermal components, or itch, contains a substance inhibiting cell adhesion between angioendothelial cells and lymphocytes. The external skin-care composition is useful for a variety of cutaneous troubles.

Description

EXTERNAL SKIN-CARE COMPOSITION

BACKGROUND OF THE INVENTION TECHNICAL FIELD

The present invention relates to an external skin-care composition which is useful in

preventing and improving a variety of cutaneous troubles typified by peeling, pachymenia, skin roughness and pigmentation.

BACKGROUND ART

In recent years, it has been a matter of significant concern to maintain healthy and beautiful skin irrespective of age or sex. However, the skin is delicately affected by

temperature, humidity, ultraviolet rays, aging, diseases, stress, eating habits and the like.

Therefore, various troubles such as the decrement of various functions of the skin and

cutaneous aging occur. The skin also undergoes various changes in appearance, which may

impose mental stress on persons. A typical aging of the skin is formation of "spots" or pigmentation. This is a beauty problem for women.

The skin is anatomically divided into epidermis, dermis and subcutaneous tissue in order from the surface thereof. The epidermis is composed mainly of keratinocytes into

which pigment cells (melanocytes), Langerhans' cells and the like are mixed. The epidermis is divided into four layers of stratum basale. stratum spinosum, stratum granulosum and

stratum comeum under an optical microscope. The keratinocytes divide at an almost constant rate in the stratum basale. which is the lowest stratum, and are successively pushed up from

the stratum basale (keratinization). The keratinocytes gradually flatten during this process

and have a thin, tabular structure at the stratum comeum. The outermost stratum comeum peels as so-called dirt

On the other hand, fibroblasts and mast cells exist in the dermis in a form where they

are surrounded by proteins called the extracellular matrix, such as collagen and elastin

Besides, a great number of minute vessels enter the dermis

In the skm, the cells and proteins as descπbed above maintain healthy and beautiful

skin in close cooperation with each other However, the skin gradually undergoes changes in

its function, properties, structure, appearance and the like due to agmg and external

environments, and so it may become hypersensitive or feel itchy from the viewpoint of

sensation and may have an increase in spots (pigmentation), disorder of skin texture or skm

roughness based on keratonosis from the viewpoint of appearance These various phenomena

are different from diseases and do not interfere with everyday life by themselves However, it is an extremely important matter from a beauty or mental point of view Everybody hopes to

maintain a fresh and healthy skin forever

The usual skin irritation and itch may be caused by various factors, for example,

soaps, shampoo, rinse, shaving, rubbing, drying, temperature changes, sunlight, etc , but are

scarcely said to be diseases Therefore, they have not been regarded as important until now

Antihistam es and steroids heretofore m use as drugs, and antipyretic-analgesic agents such

as lbuprofen are only used for excessive itch and fever It has recently been reported that

substance P antagonists are effective for allergic skin (Japanese Patent Application Laid-Open

No 304649/1995)

There is no strict definition of "skin texture' However, the disorder thereof can be

generally understood to be difierent trom wrinkles in which deep irregularities are recognized

on the skin in appearance, and cause a minute disorder of the ho y layer in the epidermis, so that the gloss and feel of the skin are affected. One cause of such disorders of the skin texture

and skin roughness based on keratinosis is also considered to be the growth and keratinization

cycle of epidermal corneocytes which is changed by some endogenous factor of the vital

body or environmental factor as described above.

The mechanism of cutaneous aging is not clearly known. However, since the skin is

situated at the outermost layer of the vital body and plays a role at the forefront of

biophylaxis, accumulation of damages by environmental factors greatly accelerates the cutaneous aging phenomenon. Pigmentation such as spots, which is a typical symptom of

cutaneous aging, is caused by increase in a melanotic pigment synthesized by melanocytes in the epidermis. As its causative mechanism, it is indicated that ultraviolet rays, female sex

hormones, genetic factors and the like participate therein. However, it is not yet fully

elucidated. Therefore, agents intended for prevention of melanogenesis or reduction of

existing melanin have hitherto been studied as beautifiers. There have heretofore been

proposed arbutin, kojic acid, vitamin C and licorice extracts (Japanese Patent Application

Laid-Open Nos. 23808/1988 and 149706/1989); the root of kudzu (Japanese Patent

Application Laid-Open No. 16709/1989); hydroxystilbene (Japanese Patent Application Laid-

Open No. 38009/1989); 3 -hydroxy chromone (Japanese Patent Application Laid-Open Nos.

1 1 1410/1980 and 143908/1980); and isoflavone (Japanese Patent Application Laid-Open No. 225004/1983).

The dermis situated under the epidermis is also affected by endogenous factors and

exogenous factors and undergoes disorder and/or degeneration of DNA-protein crosslinking of dermal components and crosslinking of proteins such as collagen and elastin, inactivation

of antioxidizing enzymes such as SOD (superoxide dismutase). and hyperoxidation of membranous lipids in cell components and consequent deterioration of cell function. As a result, the functions, properties and appearance of the overall skin are also affected.

Therefore, keeping the dermis healthy is believed to lead to the maintenance of healthy skin from the viewpoints of appearance and function.

SUMMARY OF THE INVENTION

It is accordingly an object of the present invention to provide an external skin-care

composition useful in preventing and improving peeling, pachymenia, skin roughness, disorder of skin texture, pigmentation, degeneration or destmction of dermal components, or

itch.

In view of the foregoing circumstances, it has been found, in the course of studying

the properties of the skin and their changes, and further the role of cell adhesion molecules in the skin, that a great amount of leukocytes, particularly, lymphocytes, may infiltrate into the

skin tissue in some cases even in skin not exhibiting the features of inflammation, such as

flare, swelling and fever, i.e., healthy skin accompanied by no inflammatory symptoms in

appearance, and the infiltrated lymphocytes exert an extremely large influence upon the skin

when formation of spots on the skin, skin roughness, peeling, pachymenia, disorder of skin

texture, and/or degeneration or destruction of dermal components occur, and also that a

substance (cell adhesion inhibiting substance), which prevents the lymphocytes from

infiltrating into the skin tissue by inhibiting cell adhesion between lymphocytes and

angioendothelial cells, can effectively prevent the formation of spots on the skin, skin

roughness, peeling, pachymenia, disorder of skin texture, and degeneration and destmction of

dermal components, thus leading to completion of the present invention.

According to the present invention. there is thus provided an external skin-care composition for preventing and improving peeling, pachymenia, skin roughness, disorder of skin texture, pigmentation, degeneration or destruction of dermal components, or itch, which

comprises a substance inhibiting cell adhesion between angioendothelial cells and lymphocytes.

According to the present invention, there is also provided use of a substance inhibiting

cell adhesion between angioendothelial cells and lymphocytes for the prevention and

improvement of peeling, pachymenia, skin roughness, disorder of skin texture, pigmentation, degeneration or destmction of dermal components, or itch.

According to the present invention, there is further provided a method of preventing and improving peeling, pachymenia, skin roughness, disorder of skin texture, pigmentation, degeneration or destmction of dermal components, or itch, by applying a composition

containing a substance inhibiting cell adhesion between angioendothelial cells and

lymphocytes to the skin to be treated.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

In recent years, studies on various diseases have been advanced at a molecular level,

and intercellular adhesion between leukocytes and angioendothelial cells or the like and infiltration of the leukocytes following this have been found to greatly participate in

immunity and inflammatory diseases. (See "Expression Regulation and Clinical Application

of Adhesion Molecule". Medical View Co., Ltd., (1991 ); Nature, Vol. 364. 149-155 ( 1993); Science. Vol. 247, 456-459 (1990): Annual Review, Immunity, 175-185 (1989); Trends in

Glycoscience and Glycotechnology, Vol. 4, No. 19. 405-414 (1992); Experimental Medicine.

Vol. 10, No. 1 1. 1402-1413 ( 1992); Experimental Medicine. Vol. 1 1 , No. 16. 2168-2175 ( 1993); Science, Vol. 255. 1 125-1 127 (1992); etc.) It has also been found that cell surface adhesion molecules such as ICAM-1 , ELAM-1 and VCAM- 1 participate in intercellular

adhesion. (See Annual Review, Immunity. 175-1 85 (1989); Infection, Inflammation,

Immunity, Vol. 19(2), 129-153 (1989): and Infection. Inflammation-Immunity, Vol. 24(3), 158- 165 (1994)).

More specifically, it has been found that when the vital body is stimulated from the

outside, a great number of leukocytes infiltrate into that site to cause the so-called

inflammatory reaction, so that upon the infiltration of the leukocytes from a vessel into tissue,

the leukocytes and angioendothelial cells adhere to each other through specific cell adhesion

molecules existing on their cell surfaces, and most of the adhered leukocytes destroy the basal

layer of the vessel as they are and exude out of the vessel. The inflammation is a local response to a lesion of a vital tissue. When the tissue is damaged by physical irritation such

as bacterial infection, injury, fever, cold, radiation and electricity, and chemical substances,

main symptoms of inflammation such as flare, swelling, fever and ache clinically appear

through the process of reproduction of chemical mediators, acceleration of vascular

permeability, migration of leukocytes such as neutrophiles and macrophages, infiltration,

granulation, tissue production and healing.

As described above, the cell adhesion between the angioendothelial cells and the

leukocytes and infiltration of the leukocytes following this have heretofore been known to be

extremely important steps in immunity and inflammatory diseases. However, it was not

known that the cell adhesion between the angioendothelial cells and the leukocytes,

particularly, lymphocytes, and the infiltration of the lymphocytes participate in various

cutaneous problems accompanied by no major symptoms of inflammation. Therefore, there has also not yet been proposed the external skin-care composition according to the present invention containing a substance (cell adhesion inhibiting substance) which can prevent the

infiltration of lymphocytes by inhibiting such cell adhesion.

The substance inhibiting cell adhesion between the angioendothelial cells and the

lymphocytes used in the present invention is not limited to any special substance. However,

nonprotein substances are particularly preferred in view of percutaneous absorption. For

example, benzoic acid anilide derivatives, gallic acid derivatives, benzene derivatives and

plant extracts are included. Further, galloyltannin and derivatives thereof, already known

peptides (J. Biological Chem.. 270(37), 21 129-21 136 (1995)), pentoxifylline (J. Invest

Dermatol. 104, 824-828 (1995)). 3-deazaadenosine (J. Immunology. 144(2), 653-661

(1990)), carboxyamide derivatives (J. Medicinal Chem., 37(6). 717-718 (1994)), inositol

polyamines (J. Biological Chem., 269(21), 15061-15066 (1994)), sialyl Lewis X and

derivatives thereof, triterpenic acid derivatives, and lactose and lactosamine derivatives may be mentioned. More preferable examples of the cell adhesion inhibiting substance may include Thujopsis dolabrata (ASUNARO) or extracts therefrom, lignans and morgins.

Thujopsis dolabrata is a genus endemic to Japan and an evergreen tree ranging in

Honshu, Shikoku and Kyushu. The wood thereof is used as a wooden basis for Wajima lacquering. However, the physiological activity of Thujopsis dolabrata has heretofore been

scarcely known though it has been reported to have an antimicrobial effect (Japanese Patent

Application Laid-Open No. 62809/1990), a beautifying effect (Japanese Patent Application

Laid-Open No. 345710/1993) and an osteopathy-preventing effect (Japanese Patent Application Laid-Open No. 340542/1994). However, it was not known that it has a cell adhesion inhibiting effect

In the present invention, a dry ground product composed mainly ol leaves, twigs and

the like (hereinafter referred to as "stock") of Thujopsis dolabrata may be used as it is It is

however preferable to use its extract The extract from Thujopsis dolabrata can be obtained

by grinding mainly leaves, twigs and the like of Thujopsis dolabrata in a dried or undπed

form, and then extracting them either with a solvent or by means of an extractor such as a Soxhlet's extractor at an ordinary temperature or an elevated temperature In the present

invention, the extract from Thujopsis dolabrata means an extract with any of various

solvents, or a dilute or concentrated solution or dry powder thereof

The extract with the solvent is obtained by extracting the stock of Thujopsis dolabrata

with water, an organic solvent (a hydrocarbon solvent such as petroleum ether, n-hexane cyclohexane, toluene or benzene, a halogenated hydrocarbon such as dichloromethane,

chloroform or carbon tetrachloride; an ether solvent such as diethyl ether, an ester solvent such as ethyl acetate, a ketone such as acetone, a basic solvent such as pyridine, a

monohydric or polyhydric alcohol solvent such as butanol. propanol. ethanol, methanol,

polyethylene glycol, propylene glycol or butylene glycol, or the like), an aqueous alcohol or

the like at usually about 3-70 °C

As a preferable specific example of a method for extracting the stock of Thujopsis dolabrata may be mentioned a method in which 100 g of a dry ground product of Thujopsis

dolabrata are immersed m 1 liter of ethanol to conduct extraction for 7 days while sometimes

stirring at room temperature, the resultant extract is filtered, and the filtrate is left at rest for 3

days at 5 °C and then filtered again, thereby obtaining a supematant

Although the above extract mav be used as an active ingredient for the cell adhesion inhibiting agent according to the present invention or other medicines in the form of a solution as extracted, it may be used in the form of a highly-active fraction by concentrating

the extract and then further treating the concentrated extract by a proper separating means, for

example, gel filtration, column chromatography on silica gel, high performance liquid

chromatography and/or the like.

Thujopsis dolabrata or the extract therefrom may be prepared into powder or paste as

it is or after diluted, concentrated or lyophilized to formulate a suitable preparation for use. It may also be used after subjecting it to purification treatments such as deodorization and

decoloring using active carbon or the like as needed.

Lignans have been found in Thujopsis dolabrata of the pine family (Chem. Pharm.

Bull., 20(6), 1 150-1 155 (1972)) and the like in plants, and various synthetic processes have been reported (Natural Product Report, 183-205 (1995); Tetrahedron Lett., 2759-2762 (1969);

Chem. Pharm. Bull., 20(6), 1150-1 155 (1972); etc.). They have heretofore been reported to

have antiviral activity and proliferation inhibiting activity against cancer cells (Planta Med.,

59, 246-249 (1993)) and inhibit bonding of PAF to platelets (Natural Product Report, 183-

205 (1995)). However, it has not been entirely known to date that they have a cell adhesion inhibiting effect.

Preferable examples of the lignans include compounds represented by the following general formula (1 ):

wherein R¹. R^: . R³ and R⁴ are the same or different from one another and individually represent a hydrogen atom, or a hydroxyl, alkyl or alkoxyl group.

In the formula (1), the alkyl group preferably has 1 -10 carbon atoms. Examples

thereof include linear or branched substituents such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl and decyl groups and also alkyl groups

substituted by a hydroxyl group, such as a hydroxyethyl group. The alkoxyl group is

preferably linear or branched and has 1-10 carbon atoms. Examples thereof include methoxy,

ethoxy, n-propyloxy, isopropyloxy, n-butyloxy, sec-butyloxy, tert-butyloxy, pentyloxy,

hexyloxy, heptyloxy, octyloxy, nonyloxy and decyloxy groups.

Of these compounds, those in which in the general formula (1 ), R¹ is a hydrogen

atom, a hydroxyl group or a short-chain alkoxyl group having 1-3 carbon atoms, and R^:, R³ and R⁴ are hydroxyl or methoxy groups are particularly preferred.

Such lignans (1 ) can be extracted from, for example, Thujopsis dolabrata (mainly

leaves and twigs thereof). The extraction is performed by extracting Thujopsis dolabrata or

dry powder thereof with water, an organic solvent (petroleum ether, n-hexane, cyclohexane. carbon tetrachloride, toluene, benzene, dichloromethane. chloroform, ether, ethyl acetate,

butanol. acetone, propanol, ethanol. methanol, pyridine, polyethylene glycol, propylene

glycol. butylene glycol or the like), an aqueous alcohol or the like at usually about 3-70°C.

As a preferable specific example of a method for extracting the stock of Thujopsis

dolabrata, may be mentioned a method in which 100 g of a dry ground product of Thujopsis

dolabrata are immersed in 1 liter of ethanol to conduct extraction for 7 days with occasional

days at 5 °C and then filtered again, thereby obtaining a supematant. The solvent is then

distilled from the resultant extract, and the resultant residue is suitably dissolved in a solvent

such as methanol, ethanol or ethyl acetate and further subjected to column chromatography using water, methanol, ethanol, ethyl acetate, chloroform, dichloromethane, hexane, acetone

or benzene as a eluting solvent and a hydrophilic polymer such as AMBERLITE XAD-2,

DIAION HP-20 or TSK gel HW-40, a SEPHADEX such as SEPHADEX LH-20, reversed

phase silica gel, silica gel, or cellulose as a support to fractionate the intended component

while identifying it by thin-layer chromatography or the like, whereby the intended product is

obtained. The product may be purified by recrystallization making use of a suitable solvent

such as benzene, ether, hexane, acetone, methanol, ethanol or water in some cases.

Various derivatives may be synthesized in accordance with the processes described in known literature (Natural Product Report, 183-205 (1995); Tetrahedron Lett., 2759-2762

(1969): Chem. Pharm. Bull., 20(6), 1 150-1 155 (1972). etc.). No particular limitation is imposed on the origin thereof.

The morgins include compounds represented by the following general formula (2):

wherein R^D and R⁽ are the same or different from each other and individually represent a hydroxyl or alkoxyl group.

Morgin is known to be contained in a madder (Chem. Pharm. Bull., 40(6), 1504-1509

(1992)). The present applicant has found that the morgin has an antimetabolic effect on

arachidic acid as its physiological effect. However, it has not been entirely known to date

that it has a cell adhesion inhibiting effect.

In the formula (2), the alkoxyl group is preferably linear or branched and has 1-6

carbon atoms. Examples thereof include methoxy. ethoxy, n-propyloxy, isopropyloxy, n-

butyloxy, sec-butyloxy, tert-butyloxy, n-pentyloxy and n-hexyloxy groups. In the present invention, those in which R⁵ is an alkoxyl group, and R⁶ is a hydroxyl group are particularly

preferred.

Such morgins (2) can be extracted from, for example, a madder. The extraction is

performed by first extracting a madder with a solvent such as diethyl ether, ethyl acetate, acetone, methanol, ethanol, hexane or water. The solvent is then distilled from the resultant

extract, and the resultant residue is suitably dissolved in a solvent such as methanol, ethanol

or ethyl acetate and further subjected to column chromatography using water, methanol. ethanol, ethyl acetate, chloroform, dichloromethane, hexane, acetone or benzene as an eluting solvent and a hydrophilic polymer such as AMBERLITE XAD-2. DIAION HP-20 or TSK

gel HW-40. a SEPHADEX such as SEPHADEX LH-20. reversed phase silica gel, silica gel,

or cellulose as a support to fractionate the intended component while identifying it by thin-

layer chromatography or the like, whereby the intended product is obtained. The product

may be purified by recrystallization making use of a suitable solvent such as benzene, ether, hexane, acetone, methanol, ethanol or water in some cases.

Various derivatives may be synthesized by suitably alkylating and acylating a morgin (R⁵ = R⁶ = OH in the formula (2)) in accordance with methods known per se in the art.

The amount of such a cell adhesion inhibiting substance to be used is preferably 0.000001 -10 wt.%, particulariy preferably 0.0001-1 wt.% based on the weight of the external

skin-care composition. Incidentally, the amount incoφorated is expressed in terms of solid

content (dry solid content) of the extract in the case of a plant extract such as Thujopsis dolabrata extract.

Into the external skin-care composition according to the present invention, may be

incoφorated additives commonly used in the classical external skin-care compositions, for

example, a water-absorbing gelling agent, a lipophilic gelling agent, a water-absorbing active ingredient, a lipophilic active ingredient, an antiseptic, an antioxidant, a solvent, a perfume

base, a filler and/or a coloring substance. An amount of these additives to be incoφorated may be an amount commonly used in the classical external skin-care compositions and may

be 0.01 -20 wt.% based on the weight of the composition.

Examples of oils which may be used include mineral oils (vaseline and the like),

vegetable oils (sunflower oil and the like), animal oils, synthetic oils (pruserin oil and the like), siiicone-containing oils and fluorinated oils (perfluoropolyether and the like). These oils may be used in combination with fatty alcohols and fatty acids (stearic acid and the like)

Examples of emulsifiers which may be used include glycerol stearate,

POLYSORBATE 60 and a mixture of PEG-6/PEG-32/glycol stearate (product of Gattefosse Co.; trade name: TEFOSE). Examples of the solvent include lower alcohols with ethanol and

isopropanol being particularly preferred. Examples of the water-absorbing gelling agent

include carboxyvinyl polymers, acrylic copolymers, polyacrylamide, polysaccharides an

natural mbber. Examples of the lipophilic gelling agent include modified clay, metal salts of fatty acids and hydrophobic silica.

Examples of the water-absorbing active ingredient include proteins, hydrolyzates of

proteins, amino acids, polyalcohols, urea, allantoin, saccharides and derivatives thereof, vitamins, and hydroxy acids. Examples of the lipophilic active ingredient include retinol and

derivative thereof, tocopherol and derivatives thereof, essential fatty acids, ceramides,

essential oils, and salicylic acid and derivatives thereof.

Examples of medicinally-effective ingredients which may be used include one or

more agents selected from antibacterial agents, antifungal agents, anti-inflammatory agents,

antipruritics, antiviral agents, keratolytic agents, free radical scavengers, antioxidants,

antisebaceous agents, antidandruff agents, antipimple agents and humectants. The

antibacterial agents include antibiotics of the clindamycin phosphate, erythromycine and tetracydine types, and the like. The antifungal agents include imidazole compounds such as

econazole, ketoconazole, miconazole and their salts; polyene compounds such as

amphotericin: allylamines such as terbinafine and octopirox; and the like. The anti¬

inflammatory agents include steroids such as hydrocortisone and betamethasone; ibuprofen and salts thereof, acetylsalicylic acid, acetoaminophen, glycyrrhetinic acid, and the like. The

antipruritics include thenaldine, trimeprazine, cyproheptadine and the like. The antiviral agents include acyclovir and the like.

The keratolytic agents include -hydroxycarboxylic acids and salts thereof, β-

hydroxycarboxylic acids and salts thereof, β-ketocarboxylic acids and salts thereof, amides,

esters, hydroxy acids such as glycolic acid, lactic acid, salicylic acid and citric acid, fruit

acids, 5-octanoyl-5-salicylic acid, and the like. The free radical scavengers include α -

tocopherol and esters thereof, superoxide dismutases, metal-chelating agents, ascorbic acid

and esters thereof, and the like.

The antisebaceous agents include progesterones and the like. The anti dandruff agents

include octopirox. zinc pyrothione and the like. The antipimple agents include retinoic acid,

benzoyl peroxide and the like. The humectants include natural and synthetic ceramides,

hyaluronic acid, cholesterol and salt thereof, collagens, and the like.

The external skin-care composition according to the present invention may be

formulated in the same form as the classical external skin-care compositions commonly used

for topical application and prepared in accordance with methods known per se in the art.

Examples of the forms of the external skin-care composition include aqueous solutions or aqueous alcohol solutions such as lotions; liquid or semiliquid emulsions of the water-in-oil

type prepared by dispersion (H/E) of a fatty phase (E) in an aqueous phase (H); liquid or semiliquid emulsions of the oil-in-water type prepared by reversed phase (E/H): cream or gel

type emulsions having soft consistency; microemuisions; and aqueous gel dispersions,

anhydrous gel dispersions and vesicular dispersions. The proportion of the fatty phase in

each emulsion is preferably 5-80 wt.%, particularly preferably 5-50 wt.% based on the weight of the external skin-care composition according to the present invention.

The external skin-care composition according to the present invention can be

formulated into a cleansing, protecting, treating or caring cream (for example, day and night

cream, cleansing cream, foundation cream or sunscreen cream) for face, hand, leg or body,

liquid foundation, cleansing lotion, skin care body lotion, sunscreen lotion, skin care lotion, gel or foam as necessary for the application intended.

EXAMPLES

The present invention will hereinafter be described by reference to the following

examples. However, the present invention is not limited to these examples.

Preparation Example 1 : Preparation (\) of Thujopsis dolabrata extract

A ground product (1kg) of Thujopsis dolabrata (a rnixture of leaves and twigs) was

immersed in 8 liters of ethanol for 7 days at ordinary temperature to extract an ethanol-

soluble component. A residue obtained by separating the resultant extract solution was then

subjected again to the same process as described above, thereby obtaining 16 liters of an

extract solution in total. The solvent in the extract solution was distilled off, and the resultant

residue was dried to solids under reduced pressure, thereby obtaining 80 g of an extract.

Preparation Example 2: Preparation (2) of Thujopsis dolabrata extract

A ground product (1kg) of Thujopsis dolabrata (a mixture of leaves and twigs) was

immersed in 8 liters of ethanol for 7 days at ordinary temperature to extract an efhanol- soluble component. A residue obtained by separating the resultant extract solution was then subjected again to the same process as described above, thereby obtaining 16 liters of an extract solution in total. After the solvent in the extract solution was distilled off to

concentrate the solution to 1 liter, 30 g of active carbon were added to the concentrated

solution, followed by stirring for 6 hours. After the thus-treated solution was deodorized and

decolored, the active carbon was separated by filtration, and the filtrate was concentrated under reduced pressure, thereby obtaining 65 g of an extract as a syrup.

Preparation Example 3: Preparation (3) of Thujopsis dolabrata extract

An operation was performed in the same manner as in Preparation Example 1 except

that 8 liters of 75% aqueous ethanol were used in place of 8 liters of ethanol, thereby

obtaining 79 g of an extract.

Preparation Example 4: Preparation (4) of Thujopsis dolabrata extract

that 8 liters of n-hexane were used in place of 8 liters of ethanol, thereby obtaining 52 g of an extract.

Preparation Example 5:

A dry ground product (180 g) of Thujopsis dolabrata was immersed in 1 liter of

ethanol to conduct extraction for 7 days while occasionally stirring the mixture at room temperature. The resultant extract solution was filtered, and the filtrate was left at rest for 3

days at 5 °C and then filtered again, thereby obtaining a supematant.

The solvent was then distilled off, and the resultant residue (2.4 g) was subjected to column chromatography (hexane/ethyl acetate, chloroform/methanol, water/methanol) and high performance liquid chromatography (YMC-ODS S-10, product of YMC Co.), thereby

obtaining 25 mg of Compound 1 (in the general formula ( 1 ) , R¹ = H, R² = R³ = R⁴ = OCH-,)_.

Preparation Example 6:

Compound 2 (in the general formula (1), R¹ = OH, R² = R³ = R⁴ = OCH₃) in an amount of 10 mg was obtained in the same manner as in Preparation Example 5.

Preparation Example 7:

Compound 3 (in the general formula (1), R¹ = R^: = R³ =R⁴= OCH₃) in an amount of

21 mg was obtained in the same manner as in Preparation Example 5.

Preparation Example 8:

Compound 2 (10 mg) obtained in Preparation Example 6 was ethylated in accordance

with a method known er se in the art. and the resultant product was purified by column

chromatography on silica gel to obtain 10 mg of Compound 4 (in the general formula (1 ) . R¹ = OC₂H₅, R³ = R⁴ = OCH₃).

Preparation Example 9:

Compound 2 (10 mg) obtained in Preparation Example 6 was propylated in

accordance with a method known per se in the art, and the resultant product was purified by column chromatography on silica gel to obtain 10 mg of Compound 5 (in the general formula

(1 ) , R¹ = OC₃H₇, R² = R³ = R⁴ = OCH₃). Preparation Example 10:

Compound 6 (in the general formula (1), R¹ = H, R2 = R⁴ = OCH₃, R³= OH) in an

amount of 15 mg was obtained in the same manner as in Preparation Example 5

Preparation Example 1 1 :

A dry madder (1 kg) was ground and extracted with 1 liter of methanol. After the

resultant extract solution was concentrated under reduced pressure, the concentrated solution was subjected to liquid-liquid partition chromatography. The resultant organic layer was

concentrated under reduced pressure and then subjected further to partition chromatography

with hexane-90% methanol. After the resultant hexane layer was concentrated and dried to

solids (16 g), the solids were subjected to column chromatography on silica gel (WAKOGEL

C-200) to obtain 250 mg of morgin (in the general formula (2), R⁵ = OCH₃, R⁶= OH).

Preparation Example 12:

Morgin (1 mmol) obtained in Preparation Example 11 was dissolved in methanol (5

ml), and 0.1N NaOCH₃ (catalytic amount) was added to the solution. The mixture was stirred

at room temperature, and the completion of the reaction was identified by TLC (thin-

layer liquid chromatography). Thereafter, an ion-exchange resin (150 mg) was added to the

reaction mixture to neutralize the mixture at room temperature. After the ion-exchange resin was separated by filtration, the filtrate was concentrated to quantitatively obtain a morgin derivative (in the general formula (2), R⁵ = R⁶ OH). Preparation Example 13:

Morgin (1 mmol) obtained in Preparation Example 1 1 .vas dissolved in

dimethylformamide (DMF, 3 ml), and 1.1 mmol of CH₃I and 2 mmol of CaC0₃ were added

to the solution, thereby methylating the hydroxyl group. After the completion of the reaction

was identified by TLC. extraction was conducted with CHC1₃-H₂0 (H\ OH^"), and the extract was subjected to column chromatography on silica gel, thereby obtaining 270 mg of a morgin derivative (in the general formula (2), R⁵ = R⁶ = OCH₃.

Preparation Example 14:

The morgin derivative (1 mmol) obtained in Preparation Example 12 was dissolved in

methanol (5 ml), and the solution was stirred at room temperature in the presence of WSC

(water-soluble carbodiimide, 2 mmol). After the completion of the reaction was identified by

TLC, extraction was conducted with CHC1₃-H₂0 to obtain 272 mg of a morgin derivative (in

the general formula (2), R⁵ = OC₂H₅, R⁶ = OH).

Test Example 1 : Adhesion inhibiting test between leukocvtes and angioendothelial cells

Each test substance was added to human angioendothelial cells turned confluent on a

96-well culture plate so as to give a final concentration of 0.0001 wt.%. After 18 hours,

human IL-1 α was added so as to give a final concentration of 2.5 ng/ml. followed by culture

for 6 hours. After the culture solution was removed, the plate was washed twice with RPM1- 1640 medium, and 200 μl of human peripheral leukocytes (I O⁶ cells/ml) labeled with ¹Cr in

advance were then added to conduct culture. After 30 minutes, nonadhered cells were

removed, and adhered cells were lysed. followed bv measurement of radioactivity. As a result. Thujopsis dolabrata extracts tested were found to have excellent inhibitory activity against cell adhesion between the leukocytes and the angioendothelial cells

as shown in Table 1

Table 1

Test substance Inhibition rate of leukocyte adhesion (%)

Thujopsis dolabrata extract with ethanol 85

Thujopsis dolabrata extract with hexane 90 Thujopsis dolabrata extract with ethyl acetate 86

Test Example 2' Toxicity test against angioendothelial cell (cell form. DNA synthesis^

With respect to moφhological changes, judgment was conducted visually through an inverted microscope. On the other hand, DNA synthesis was evaluated by using uptake of

3H-thymidine as an index in accordance with a method known per se in the art and

determining the uptake rate for the late 8 hours in a 24-hour culture after adding a test

substance by means of a liquid scintillation counter. The concentration of the test substance was controlled to 0.0001 wt.%.

As a result, all Thujopsis dolabrata extracts tested were found to have low toxicity against angioendothelial cells as shown in Table 2

Table 2

Test substance Moφhological change Inhibition rate of DNA synthesis (%)

Thujopsis dolabrata extract with ethanol Underwent no particular change 10

Thujopsis dolabrata extract with hexane Underwent no particular change 11

Thujopsis dolabrata extract with ethyl Underwent no particular change acetate

7

Test Example 3 : Adhesion inhibiting test between leukocvtes and angioendothelial cells

96-well culture plate so as to give a final concentration of 10 nM. After 3 hours, human IL-1

α was added so as to give a final concentration of 2.5 ng/ml, followed by culture for 6 hours.

After the culture solution was removed, the plate was washed twice with RPMI- 1640

medium, and 200 μl of human peripheral leukocytes (IO⁶ cells/ml) labeled with ^{5 l}Cr in

removed, and adhered cells were lysed in 0.1% SDS/50 mM TRIS solution, followed by

measurement of radioactivity.

As a result, the lignans tested were found to have excellent inhibitory activity against

cell adhesion between the leukocytes and the angioendothelial cells as shown in Table 3.

Test Example 4: Toxicity test against angioendothelial cell (protein s^ynthesis)

Protein synthesis was evaluated by using uptake of ³H-leucine as an index in

accordance with a method known per se in the art and determining the uptake rate for the late

4 hours in a 24-hour culture after adding a test substance by means of a liquid scintillation i counter. The concentration of the test substance was controlled to 10 nM.

As a result, all the lignans tested were found to have low toxicity against

angioendothelial cells as shown in Table 3.

Table 3

Test Formula (1) Inhibition Inhibition substance rate of rate of leukocyte protein

R¹ R² R³ R⁴ adhesion synthesis

(%) ^' (%)

Compd. 1 H OCH₃ OCH₃ OCH₃ 80 8

Co pd. 2 OH OCH₃ OCH₃ OCH₃ 83 6

Compd. 3 OCH₃ OCH₃ OCH₃ OCH₃ 74 1

Compd. 4 OC₂H₅ OCH₃ OCH₃ OCH₃ 75 4

Compd. 5 OC₃H₇ OCH₃ OCH₃ OCH₃ 72 6

Compd. 6 H OCH₃ OH OCH₃ 84 6

Compd. 7 OH OCH₃ OH OCH₃ 86 10

Compd. 8 OCH₃ OCH₃ OH OCH₃ 80 2

Test Example 5: Adhesion inhibiting test between leukocvtes and angioendothelial cells

Each test substance was added to human angioendothelial cells turned confluent on a 96-well culture plate so as to give a final concentration of 10 μM. After 18 hours, human IL-

1 α was added so as to give a final concentration of 2.5 ng/ml, followed by culture for 6 hours. After the culture solution was removed, the plate was washed twice with RPMI- 1640

medium, and 200 μl of human peripheral leukocytes (IO⁶ cells/ml) labeled with Cr in

removed, and adhered cells were lysed in 0.1 % SDS/50 mM TRIS solution, followed bv measurement of radioactivity.

As a result, the morgin and derivatives thereof tested were found to have excellent

inhibitory activity against cell adhesion between the leukocytes and the angioendothelial cells as shown in Table 4.

Test Example 6: Toxicity test against angioendothelial cell (cell form. DNA synthesis

With respect to moφhological changes, judgment was conducted visually through an

inverted microscope. On the other hand, DNA synthesis was evaluated by using uptake of

H-thymidine as an index in accordance with a method known per se in the art and

determining the uptake rate for the late 4 hours in a 24-hour culture after adding a test

substance by means of a liquid scintillation counter. The concentration of the test substance

was controlled to 10 μM.

As a result, all the morgin and derivatives thereof tested were found to have low

toxicity against angioendothelial cells as shown in Table 4.

Table 4

Test Formula (2) Inhibition Inhibition Moφhological substance rate of rate of change of leukocyte DNA angioendo¬

R⁵ R⁶ adhesion synthesis thelial cell

(%) ^' (%)

Prepn. Ex. 1 1 OCH₃ OH 94 5 Not changed

Prepn. Ex. 12 OH OH 54 7 Not changed

Prepn. Ex. 13 OCH₃ OCH, 88 3 Not changed

Prepn. Ex. 14 OC_:H₅ OH 93 5 Not changed Test Example 6: Effect of cell adhesion inhibiting substance on peeling

After shaving the backs of white guinea pigs, a cell adhesion inhibiting substance (100 mM; 10 μl) was applied to 1 cm² of each of their backs. After 2 hours, a seal for a patch

test, in which a 1% SDS (sodium dodecyl sulfate) solution had been soaked, was applied

thereto for 30 minutes. Thereafter, the test substance was applied again. This process was

repeated for 7 days. After 7 days, the thus-treated skins were collected, H-E staining was conducted, and photographs were taken through a microscope. Thereafter, pachymenia and

peeled state of the epidermides were determined and observed. The thickness of each

epidermis was expressed in terms of actual measurements, while the degree of peeling was visually evaluated by relative comparison and ranked in 4 grades by which the state of a

control untreated with SDS was ranked as 1, and the state of a sample treated with SDS was ranked as 4. The results are shown in Table 5.

Table 5

Sample applied Thickness of epidermis (mm) Degree of peeling

SDS + ethanol 39 4

Control (untreated with SDS) 15 1

SDS + butyl gallate 19* 2

SDS + benzoic acid anilide 22 2

*P < 0.05

Test Example 7: Effect of cell adhesion inhibiting substance on disorder of skin texture

A cell adhesion inhibiting substance (0.1 %; 15 μ l ) was applied to 1 cm² of a human

skin. After 2 hours, dry nitrogen gas was blown on the applied site for 30 minutes. This process was repeated for 10 days. After 10 days, a skin replica was collected from the thus- treated skin using a HYDROPHILIC EXAFLEX hydrophilic vinylsilicone impression

material (product of D C K.K.). The condition of the skin texture was visually judged from

this replica and ranked in 5 grades from 1 where the skin texture was poor to 5 where the skin texture was good. The results are shown in Table 6.

Table 6

Sample applied Condition of skin texture

Control (normal skin) 4.2

Only nitrogen gas 2.1

Nitrogen gas + Thujopsis dolabrata extract 2.8 (of Preparation Example 2)

Nitrogen gas + morgin 3.0

Test Example 8: Effect of cell adhesion inhibiting substance on itch

A cell adhesion inhibiting substance (100 mM; 15 μl ) was applied to the backs of

Balb/c mice. After 2 hours, a histamine solution was intracutaneously injected into the mice.

Thereafter, the behavior of the mice was recorded over 1 hour by a video camera. The

amount of time the mice were scratching their sites dosed with histamine was counted from the video image to evaluate the effect on itch. The results are shown in Table 7.

Table 7

Sample applied Amount of time scratched (sec)

Not applied (dosed with histamine) 70

Control (undosed with histamine) 33

Butyl gall ate 46*

Galloylglucose 52*

*P < 0.05

Test Example 9: Effect of cell adhesion inhibiting substance on pigmentation

After carefully shaving the back fur of brown guinea pigs with a hair clipper and a shaver, each back was divided into 8 portions each having an area of 1.5 cm². The back was

exposed to ultraviolet rays in a UVB region in an intensity of 0.3 times as much as a minimal

erythema dose (MED) in order to facilitate pigmentation. After the UVB irradiation, a sample (0.1-1 % ethanol solution; 20 μl) to be evaluated was applied to each of the divided

portions. After this process was repeated for 7 days, only the sample was repeatedly applied

for 1 week. After 2 weeks, the degree of pigmentation was determined. Ethanol was applied

in place of the sample as a control to evaluate the effect of the sample. The results are shown in Table 8.

The evaluation was performed by conducting measurement by means of a calorimeter,

and calculating an L* value from the thus-obtained Munsell value to find a value ( L*) by

subtracting an L* value before the UVB irradiation from an L* value after 2 weeks from the

beginning of the UVB irradiation, thereby comparing the degree of pigmentation. The greater -ΔL* value indicates that the degree of pigmentation is more intense. ΔL* = (L* value after UVB irradiation) (L* value before UVB irradiation)

Table 8

Sample applied -ΔL* value

Ethanol (control) 5.7

Morgin 3.3*

Benzoic acid anilide 3.0*

*P < 0.05

Test Example 10: Effect of cell adhesion inhibiting substance on destmction of dermal components

Backs of hairless rats were exposed to ultraviolet rays in a UVB region at an intensity

of 0.3 times as much as a minimal erythema dose (MED) in order to accelerate the

degeneration of dermal components. A sample (0.1-1% ethanol solution; 20 μl per cm²) to be

evaluated was applied to each of the thus-exposed backs of the rats. After this process was

repeated for 20 days, the dermides were collected to conduct Masson's trichrome stain,

thereby analyzing a proportion of collagen fibers stained blue per area unit by means of an

image analyzer. The proportion of bluing per unit area becomes lower as the degree of

degeneration of collagen fibers is higher. Ethanol was applied in place of the sample as a

control to evaluate the effect of the sample. The results are shown in Table 9.

Table 9

Sample applied Degree of bluing (%)

Ethanol (control) 65

Galloylglucose 74*

Benzoic acid anilide 72*

Thujopsis dolabrata extract (of 70 Preparation Example 2)

Unexposed to UVB 85

* P < 0.05

External skin-care compositions according to Examples 1-19 were prepared in

accordance with a method known per se in the art.

Example 1 : Skin lotion

(wt.%)

Cell adhesion inhibiting substance 0.5

(benzoic acid anilide) Glyceryl monostearate 1

Ethanol 15

Propylene glycol 4

Isopropyl palmitate 3

Lanolin 1 1

Methyl p-hydroxybenzoate 0.1

Perfume base, coloring matter Trace amount

Purified water Balance Example 2: Pack composition

(wt.%)

Cell adhesion inhibiting substance 0.9

(pentoxifylline)

Polyvinyl alcohol 20 Glycerol 5 Ethanol 16

Perfume base, coloring matter Trace amount Purified water Balance

Example 3: Face-care gel

(wt.%)

Cell adhesion inhibiting substance 1

(Thujopsis dolabrata extract (of Preparation

Example 3))

Hydroxypropyl cellulose Antioxidant 0.05 Isopropanol 40 Antiseptic 0.3 Purified water Balance

Example 4: Face-care cream

(wt.%) Cell adhesion inhibiting substance 0.2

(Sialyl Lewis X)

Squalane

Stearic acid

Glyceryl monostearate 10 Ethanol

Methyl p-hydroxybenzoate 0.2 Cetanol

Olive oil Vaseline

Natural ceramide 1 Perfume base, coloring matter Trace amount Purified water Balance

Example 5: Toilet lotion

(wt.%)

Cell adhesion inhibiting substance 0.05

(Butyl gallate)

1,3-Butylene glycol 6.5 Polyoxyethylene sorbitan monolaurate

Ethanol 8 Antiseptic Trace amount Perfume base Trace amount Purified water Balance

Example 6: Cream

(wt.%)

Cell adhesion inhibiting substance 0.05

(3-deazaadenosine)

Cholesterol 0.5 Cholesterol isostearate 1 Polyether-modified silicone 1.5 Cyclic silicone 20 Methylphenylpolysiloxane 2 Methylpolysiloxane 2 Magnesium sulfate 0.5 Ethanol J

C arboxymethy 1 chitin 0.5 Perfume base, coloring matter Trace amount Purified water Balance

Example 7: Cream (oil-in-water type emulsion)

(wt.%)

Cell adhesion inhibiting substance 0.05 (methyl gallate)

Glyceryl stearate TWEEN 60

Stearic acid 1 4

Triethanolamine 0 7

Carbomer 0 4

Liquid component (Waxes, oils and esters) 1 1 5

Perhydrosqualene 12.5

Antioxidant 0 04

Perfume base Trace amount

Antiseptic Trace amount

Purified water Balance

Example 8' Toilet lotion

(wt %)

Cell adhesion inhibiting substance 0.05

(Thujopsis dolabrata extract (of

Preparation Example 2))

10-Hydroxyundecanoic acid 0.75

9-Hydroxyundecanoic acid 0.20

8-Hydroxyundecanoic acid 0.05

Arginine 0 4

Lysine 0 4

Polyoxyethylene hardened 1 5 castor oil (40 EO)

J J Methylpolysiloxane.methyl- 0.6 (polyoxyethylene)siloxane copolymer

(SH3775C, product of Dow Coming Toray

Silicone Co., Ltd.)

Glycerol 5.0

1,3-Butylene glycol 3.0

Glycinebetaine 0.2

Trisodium citrate 0.9

Citric acid 0.4

Urea 0.5 ε-Aminocaproic acid 0.1

Ethanol 5.0

Antiseptic q.s.

Perfume base q.s.

Purified water Balance

Example 9: Toilet lotion

(wt.%)

Cell adhesion inhibiting substance 0.50

(morgin) 10-Hydroxyundecanoic acid 0.89 9-Hydroxyundecanoic acid 0.20 8-Hydroxyundecanoic acid 0.02 Triethanolamine 0.4

Potassium hydroxide 0.2

Polyoxyethylene isocetyl ether (20 EO) 0.3

Sodium polyoxyethylene oleyl ether 0.3

phosphate (8 EO)

Sodium polyoxyethylene dialkyl 0.1

phosphate (10 EO) Polyethylene glycol 1500 2.0

Polyoxyethylene methylglucoside ( 10 EO) 1.5

Dipropylene glycol 0.5

Disodium hydrogenphosphate 0.5

Succinic acid 0.3

Ethanol 10.0

Royal jelly extract 2.0

Antiseptic q.s.

Perfume base q.s.

Purified water Balance

Example 10: Cosmetic jellv

(wt.%) Cell adhesion inhibiting substance 0.5

(morgin) 10-Hydroxyundecanoic acid 1.7

J 9-Hydroxyundecanoic acid 0.2

8-Hydroxyundecanoic acid 0.1

Polyoxyethylene isocetyl ether (20 EO) 1 .0

Sodium polyoxyethylene trialkyl 1 .0

phosphate (10 EO)

Sorbitol 0.5

1 ,3-Propanediol 0.5

Xanthan gum 0.5

Tuberose polysaccharide 3.0

Carboxyvinyl polymer (CARBOPOL 940, 0.2

product of Goodrich Company)

Dipotassium glycyrrhetinate 0.1

Potassium hydroxide 0.9

Allantoin 0.1

Tannic acid 0.2

Ethanol 20.0

Antiseptic q.s.

Perfume base q.s.

Purified water Balance

Example 1 1 : Cosmetic jellv

(wt.%)

Cell adhesion inhibiting substance 0.5 (Thujopsis dolabrata extract (of

Preparation Example 3)) 10-Hydroxyundecanoic acid 1.5

9-Hydroxyundecanoic acid 0.4

8-Hydroxyundecanoic acid 0.1

Silicone composition (KSG-17, product 5.0

of Shin-Etsu Chemical Co., Ltd.)

Methylpolysiloxane (KF96A-6cs, product 15.0

of Shin-Etsu Chemical Co., Ltd.)

Methylpolysiloxane (SH244, product of 5.0

Dow Corning Toray Silicone Co., Ltd.)

Methylpolysiloxane,methyl(polyoxy- 2.0

ethylene)siloxane copolymer (SH3771C, product of Dow Corning Toray Silicone

Co., Ltd.)

Methylpolysiloxane,methyl(polyoxy- 2.0

efhylene)siloxane copolymer (SH3775C, product of Dow Corning Toray Silicone

Co., Ltd.)

Methyl cellulose 0.2

Hydroxyethyl cellulose hydroxypropyl- 0.02

trimethylammonium chloride ether

(CATICELLO H-60, product of Kao Coφoration)

dl- α -Tocopherol acetate 0.05

Isostearyl glycyrrhetinate 0.1

Isopropylmethylphenol 0.1

EDTA 0.1

Ethanol 5.0

Antiseptic q.s.

Perfume base q.s.

Purified water Balance

Example 12: Cosmetic emulsion

(wt.)

Cell adhesion inhibiting substance 0.1

(ethyl gallate)

10-Hydroxyundecanoic acid 1.07

9-Hydroxyundecanoic acid 0.36

8-Hydroxyundecanoic acid 0.07

N-(3-Hexadecyloxy-2-hydroxypropyl)-N-2- 0.5 hydroxyethyldecan amide

N-(3-Hexadecyloxy-2-hydroxypropyl)-N-2- 1 .0 hydroxyethylhexadecanamide

N-(3-Tetradecyloxy-2-hydroxypropyl)-N-2- 0.5 bydroxyethyldecanamide Polyoxyethylene hardened castor oil 1.0

( 10 EO)

Methylpolysiloxane,methyl(poIyoxy- 1.0

ethylene)siloxane copolymer (SH3775C,

product of Dow Corning Toray Silicone

Co., Ltd.)

Sorbitan monostearate 0.2

Sodium stearoylmethyltaurine 0.5

Cholesterol 0.8

Cholesterol isostearate 0.2

Monocholesteryl alkenylsuccinate 0.8

Stearic acid 0.2

Palmitic acid 0.3

Myristic acid 0.1

Glycol pentyl dicaprate 4.0

Methylpolysiloxane (KF96A-500cs, product 2.0

of Shin-Etsu Chemical Co., Ltd.)

Isostearyl alcohol 1.2

Cetyl alcohol 1.0

Glycerol 3.5

Lactic acid 0.2

Sodium lactate 0.3

Natural ceramide 0.3 Antiseptic q.s. Perfume base q.s. Purified water Balance

Example 13: Toilet lotion

(wt.%)

Cell adhesion inhibiting substance 0.50

(morgin) 7-Hydroxyoctanoic acid 0.75 7-Hydroxynonanoic acid 0.20 7-Hydroxydecanoic acid 0.05 Arginine 0.4

Lysine 0.4 Polyoxyethylene hardened castor oil 1.5

(40 EO)

Methylpolysiloxane,methyl(polyoxy- 0.6

ethylene)siloxane copolymer (SH3775C,

product of Dow Coming Toray Silicone

Co., Ltd.)

Maltitol 5.0

Ethylene glycol monoethyl ether 0.2

Trisodium citrate 0.9

Succinic acid 0.4 Urea 0.5 ε-Aminocaproic acid 0.1

Ethanol 5.0

Dipotassium glycyrrhetinate 0.01

Antiseptic q.s.

Perfume base q.s.

Purified water Balance

Example 14: Toilet lotion

(wt.%)

Cell adhesion inhibiting substance 0.50

(benzoic acid anilide)

1 1-Hydroxydodecanoic acid 0.73

1 1-Hydroxytridecanoic acid 0.32

1 1 -Hydroxytetradecanoic acid 0.03

Diisopropanolamine 0.4

Potassium hydroxide 0.2

Polyoxyethylene isocetyl ether (20 EO) 0.3

Sodium polyoxyethylene oleyl ether 0.3 phosphate (8 EO)

Sodium polyoxyethylene dialkyl phosphate 0.1

(10 EO)

Polyethylene glycol 1500 2.0 Polyoxyethykne methylglucoside (20 EO) 1 _.5

Isoprene glycol 0.5

Disodium hydrogenphosphate 0.5

Succinic acid 0.3

Ethanol 10.0 Yeast extract 2.0

Antiseptic q.s.

Perfume base q.s.

Purified water Balance

Example 15: Cosmetic jellv

(wt.%) Cell adhesion inhibiting substance 0.5

(Thujopsis dolabrata extract (of

Preparation Example 3))

Ethyl 10-hydroxyundecanoate 1.9

Glyceryl 9-hydroxyundecanoate 0.2

Hydroxyethyl 8-hydroxyundecanoate 0.1

Polyoxyethylene isocetyl ether (20 EO) 1.0

Sodium polyoxyethylene trialkyl 1.0

phosphate (10 EO)

Sorbitol 0.5

1 ,3-Propanediol 0.5 Xanthan gum 0.5

Tuberose polysaccharide 3.0

Carboxyvinyl polymer (CARBOPOL 940, 0.2

product of Goodrich Company)

Potassium hydroxide 0.9

Allantoin 0.1

Zinc sulfocarbolate 0.2 Ethanol 20.0 Antiseptic q.s. Perfume base q.s. Purified water Balance

Example 16: Cosmetic jellv

(wt.%)

Cell adhesion inhibiting substance 0.5

(morgin)

Isopropyl 9-hydroxydecanoate 1.5 Glycine 8 -hydroxy decanoate 0.4 Ethyl 9-hydroxydodecanoate 0.1 Silicone composition (KSG-16, product 5.0 of Shin-Etsu Chemical Co., Ltd.) Methylpolysiloxane (KF96A-6cs, product 15.0 of Shin-Etsu Chemical Co., Ltd.) Methylpolysiloxane (SH244, product of 5.0

Dow Co ing Toray Silicone Co., Ltd.)

Methylpolysiloxane,methyl(polyoxy- 2.0

ethylenejsiloxane copolymer (SH3771C,

product of Dow Corning Toray Silicone

Co., Ltd.)

Mefhylpolysiloxane,methyl(polyoxy- 1.0

ethylenejsiloxane copolymer (SH3775C, product of Dow Coming Toray Silicone

Co., Ltd.)

Methyl cellulose 0.2

Hydroxyethyl cellulose hydroxypropyl- 0.02 trimethylammonium chloride ether

(CATICELLO H-60, product of Kao

Coφoration) dl- a -Tocopherol acetate 0.05

Isostearyl glycyrrhetinate 0.1

lsopropylmethylphenol 0.1

Zinc white 1.5

Ethanol 5.0

Antiseptic q.s.

Perfume base q.s.

Purified water Balance Example 1 7: Cosmetic emulsion

(wt.%)

Cell adhesion inhibiting substance 0.5

(3 -deazaadenosine) Isostearyl 10-hydroxyundecanoate 1.25

Ethyl 9-hydroxyundecanoate 0.46

Glyceryl 8-hydroxyundecanoate 0.09

N-(3-Hexadecyloxy-2-hydroxypropyl)-N-2- 0.5

hydroxyethyldecanamide N-(3-Hexadecyloxy-2-hydroxypropyl)-N-2- 1.0

hydroxyethylhexadecan amide

N-(3-Tetradecyloxy-2-hydroxypropyl)-N-2- 0.5

hydroxyethyldecanamide

Polyoxyethylene hardened castor oil 1.0

(10 EO)

Methylpolysiloxane,methyl(polyoxy- 1.0 ethylene)siloxane copolymer (SH3773C,

product of Dow Coming Toray Silicone

Co., Ltd.)

Sorbitan monostearate 0.2

Cholesterol 0.8

Cholesterol isostearate 0.2

Monocholestervl alkenvlsuccinate 0.8 Stearic acid 0.5

Squalane 4.0

Methylpolysiloxane (KF96A-500cs, product 2.0 of Shin-Etsu Chemical Co., Ltd.)

2-Ethylhexyl p-methoxycinnamate (PERSOLE 2.0

MCX-SA, product of Givaudan K.K.) Cetyl alcohol 1.0 Glycerol 3.5 Lactic acid 0.2 Sodium lactate 0.3 Antiseptic q.s. Perfume base q.s. Purified water Balance

Example 18: Pack

(wt.%)

Cell adhesion inhibiting substance 1.0

(galloylglucose)

10-Hydroxyundecanoic acid 4.0 9-Hydroxyundecanoic acid 0.85 8-Hydroxyundecanoic acid 0.15 Polyvinyl alcohol (GOSENOL EG-30, 15.0 product of The Nippon Synthetic Chemical Industry Co , Ltd.) Aqueous solution of carboxymelhylchithin 5.0

(CHITHIN LIQUID HV-10. product of

Ichimaru Pharcos Co., Ltd.) Triglucopolysaccharide (PULLULAN PL-20, 0.5

product of Hayashibara Company, Ltd.)

Xanthan gum 0.5

Sodium carboxymethyl cellulose 0.5

Titanium oxide 15.0

Aluminum magnesium silicate 1.0 l-Isostearoyl-3-myristoyl-glγcerol 1.0

Diglycerol 1.5

Polyoxyethylene isocetyl ether (20 EO) 1.0

Ethanol 5.0

Antiseptic q.s.

Perfume base q.s.

Purified water Balance

Example 19: Pack

(wt.%) Cell adhesion inhibiting substance 0.5

(3 -deazaadenosine)

10-Hydroxyundecanoic acid 4.0 10-FIydroxydodecanoic acid 0.85

7-Hydroxydecanoic acid 0.15

Polyvinyl alcohol (GOSENOL EG-30, 15.0

product of The Nippon Synthetic

Chemical Industry Co., Ltd.)

Aqueous solution of carboxvmethylchithin 5.0

(CHITHIN LIQUID HV-10, product of

Ichimaru Pharcos Co., Ltd.)

Triglucopolysaccharide (PULLULAN PL-20, 0.5

product of Hayashibara Company, Ltd.)

Xanthan gum 0.5

Sodium carboxymethyl cellulose 0.5

Titanium oxide 15.0

Aluminum magnesium silicate 1.0

1 -Isostearoyl-3-myristoyl-glycerol 1.0

Diglycerol 1.5

Polyoxyethylene isocetyl ether (20 EO) 1.0

Ethanol 5.0

Antiseptic q.s.

Perfume base q.s.

Purified water Balance INDUSTRIAL APPLICABILITY

The external skin-care compositions according to the present invention are useful for

preventing and improving peeling, pachymenia, skin roughness, disorders of skin texture,

pigmentation, degeneration and destmction of dermal components, or itch.

Obviously, numerous modifications and variations of the present invention are possible

in light of the above teachings. It is therefore to be understood that within the scope of the

appended claims, the invention may be practiced otherwise than as specifically described

herein.

Claims

1. An external skin-care composition for preventing and improving peeling,

pachymenia, skin roughness, disorders of skin texture, pigmentation, degeneration or

destmction of dermal components, or itch, which comprises a substance inhibiting cell

adhesion between angioendothelial cells and lymphocytes and a skin-care composition base material.

2. The external skin-care composition according to Claim 1. wherein the cell adhesion

inhibiting substance is contained in an amount of 0.000001 -10 wt.% based on the weight of the

composition.

3. The external skin-care composition according to Claim 1 , wherein the cell adhesion inhibiting substance is contained in an amount of 0.0001 -1 wt.% based on the weight of the

composition.

4. The external skin-care composition according to any one of Claims 1 to 3, wherein

the cell adhesion inhibiting substance is Thujopsis dolabrata (ASUNARO) or an extract

therefrom.

5. The external skin-care composition according to any one of Ciaims 1 to 3, wherein the cell adhesion inhibiting substance is a lignan represented by the following general formula

(1):

wherein R', R², R³ and R⁴ are the same or different from one another and individually represent a hydrogen atom, or a hydroxyl, alkyl or alkoxyl group.

6. The external skin-care composition according to any one of Claims 1 to 3, wherein

the cell adhesion inhibiting substance is a morgin represented by the following general formula

(2):

wherein R⁵ and R⁶ are the same or different from each other and individually represent a

hydroxyl or alkoxyl group.

7. The external skin-care composition according to any one of Claims 1 to 6, which

further comprises one or more agents selected from antibacterial agents, antifungal agents, anti- inflammatory agents, antipmritics. antiviral agents, keratolytic agents, free radical scavengers,

antioxidants, antisebaceous agents, antidandruff agents, antipimple agents and humectants.

8. Use of a substance inhibiting cell adhesion between angioendothelial cells and lymphocytes for the prevention and improvement of peeling, pachymenia, skin roughness,

disorder of skin texture, pigmentation, degeneration or destmction of dermal components, or

itch.

9. The use according to Claim 8, wherein the cell adhesion inhibiting substance is Thujopsis dolabrata (ASUNARO) or an extract therefrom.

10. The use according to Claim 8, wherein the cell adhesion inhibiting substance is a lignan represented by the following general formula (1 ):

wherein R¹, R², R³ and R⁴ are the same or different from one another and individually represent

a hydrogen atom, or a hydroxyl, alkyl or alkoxyl group.

1 1. The use according to Claim 8, wherein the cell adhesion inhibiting substance is a

morgin represented by the following general formula (2):

hydroxyl or alkoxyl group.

12. A method of preventing and improving peeling, pachymenia, skin roughness,

itch, which comprises applying a composition containing a substance inhibiting cell adhesion

between angioendothelial cells and lymphocytes to the skin to be treated.

13. The method according to Claim 12, wherein the cell adhesion inhibiting substance

is contained in an amount of 0.000001-10 wt.% based on the weight of the composition.

14. The method according to Claim 12, wherein the cell adhesion inhibiting substance is contained in an amount of 0.0001-1 wt.% based on the weight of the composition.

15. The method according to Claim 12, wherein the cell adhesion inhibiting substance is Thujopsis dolabrata (ASUNARO) or an extract therefrom.

16. The method according to Claim 12, wherein the cell adhesion inhibiting substance

is a lignan represented by the following general formula (1):

wherein R¹, R², R³ and R⁴ are the same or different from one another and individually represent a hydrogen atom, or a hydroxyl, alkyl or alkoxyl group.

17. The method according to Claim 12, wherein the cell adhesion inhibiting substance is a morgin represented by the following general formula (2):

hydroxyl or alkoxyl group.