WO1987007505A1

WO1987007505A1 - Wound-healing drug and cosmetics

Info

Publication number: WO1987007505A1
Application number: PCT/JP1987/000364
Authority: WO
Inventors: Masakazu Adachi
Original assignee: Japan Immuno Research Laboratories Co., Ltd.; Otsuka Pharmaceutical Co., Ltd.
Priority date: 1986-06-13
Filing date: 1987-06-09
Publication date: 1987-12-17
Also published as: KR880013570A; JPS63107912A; JPH0669958B2; JPH05306234A; JPH0529333B2; KR940003055B1; JPH0672086B2; JPH05246884A

Abstract

A wound-healing drug, an anti-inflammatory drug, a hair tonic, cosmetics, etc. containing ovomacroglobulin as an effective ingredient. They act on the skin, head skin, hair, etc. to show an excellent wound-healing effect, a hair-growing effect, and a skin- and hair-protecting effect.

Description

Details

Wound healing agent and cosmetics

Technical field

The present invention relates to a wound healing agent, a hair follicle> a cosmetic, and more particularly, to a wound healing agent characterized by containing ovamacroglobulin as an active ingredient. The present invention relates to pharmaceuticals or cosmetics.

Background art

In the treatment of wounds, for example, general trauma, hemorrhoids, pressure sores, etc. and wounds due to major surgery that extends to a deeper part of the body, promotion of granulation at the damaged part in the process And promotion of epidermis formation are important issues ο

At present, compounds that can be used for the treatment of wounds include retinoic acid, allantoin, asiaceous icoside (Aciaceae), and asia at icosi de.銪 etc. are known. And force, while these drugs are displaced also for the stomach ₀ or even still be sufficiently exhibit the action of promoting the granulation and the like, scan terrorism Lee de system during thermal wounds, non-scan terrorist Lee de It is known that the use of anti-inflammatory drugs in the system rather abolishes the resistance]. However, when these drugs are administered, it is necessary to use them in combination with drugs that can activate and enhance the resistance. Is done.

Furthermore, as a drug that has recently been effective in treating wounds, it is isolated from mammalian body fluids and has a molecular weight of about 5300 and is composed of 52 amino acids. Japanese Patent Application Laid-Open No. 57-37871 16) and compositions containing essential amino acids In addition, an egg containing iodine at a high degree of 300 U above 300 U ^ has been proposed to promote wound treatment and reduce muscular disease. It is effective for prevention (Japanese Patent Application Laid-Open No. 59-116162), and a skin cosmetic comprising dried egg white as a base for skin cosmetics (Japanese Patent Publication No. Sho 61-125) No. 6801) has also been proposed, but those which can provide sufficient effects for the treatment of wounds are also possible. O In addition, the whole eggs, egg whites, etc., are used in the presence of a preservative. The formulation containing these had a problem with the storage stability.o In addition, egg whites would not dissolve unless they were alkaline, and wound preparations. It has low affinity with the constituent materials of the softeners and cosmetics, and easily produces a white precipitate containing the protein.

On the other hand, various hair restorers and so-called hair follicles that act on the hair root and promote hair growth have been provided, but those that are still sufficiently effective have not been obtained.

Furthermore, it has an excellent protective effect on skin against rough skin, etc., as well as a protective effect on the skin (lubrication assurance), a treating effect and an excellent affinity. O Development of cosmetics was desired

Disclosure of the invention

SUMMARY OF THE INVENTION The inventors of the present invention have studied diligently, Gogo and Oboma globulin, which have excellent wound healing promotion effects, hair growth promotion effects, skin protection effects, and the like. Wound healing agent, hair restorer, hair restorer with excellent action The present inventors have found that hair fly plans and cosmetics can be obtained, and have completed the present invention.

That is, the object of the present invention is to provide a wound treatment agent, hair fly plan, cosmetics, etc. containing omaclog π-brin as an active ingredient.

Another object of the present invention is to provide a method for treating a wound, which is characterized by applying Ovo macroglobulin to an affected area.

Still another object of the present invention is to provide a hair growth promoting method characterized by applying ovomacroglobulin to scalp and hair. .

Still another object of the present invention is to provide a skin protection method characterized by applying Oboma Gloprolin to the skin.

Brief description of drawings

Fig. 1 is a graph showing the change in body weight of a test mouse in a burn recovery test using a burn wound mouse.o Fig. 2 is an inflammatory model to which the therapeutic agent of the present invention was applied. Fig. 3 is a drawing showing a cross-section of the mouse group after the injury β day]), and Fig. 3 is a drawing showing a cross-sectional view of the control mouse to which the hydrophilic ointment was administered.

FIG. 4 is a drawing showing the appearance of a wounded part of a mouse with a burn wound to which the therapeutic agent of the present invention has been applied 6 days later, and FIG. 5 is a drawing showing the appearance of the wounded part of a mouse with a control group.

FIG. 6 shows a burn-injured mouse to which the therapeutic agent of the present invention was applied. Drawing showing the cross section of the tissue 4 days after the injury to the patient, and Fig. 7 is a drawing showing the new surface of the mouse in the control group o

FIGS. 8 and 9 are drawings showing the appearance of the useful part of the skin-injured mouse to which the therapeutic agent of the present invention was applied 7 days after the injury. D »FIG. 10 O is the mouse in the control group. It is a drawing showing the appearance of the damaged part o

Fig. 11 and Fig. 12 are drawings showing the appearance of the injured area of the wounded mouse on which the therapeutic agent of the present invention was applied 21 days after the injury. Fig. 13 is a mouse in the control group. O This is a drawing showing the external appearance of the damaged part o

FIG. 14 is a drawing showing extension of corneal epithelial cells. FIG. 15 is a drawing showing a tissue section of a periodontal tissue-injured mouse to which the therapeutic agent of the present invention was applied 7 days after the injury! ), Fig. 16 is a drawing showing the cross section of the mouse of the control group o The best mode for carrying out the invention

Examples of the wound treatment agent of the present invention include: ? Wound, cut! ) Wounds, burns, burns, frostbite, skin ulcers, dry skin, skin; * keratosis, cracks, redness, dermatitis, sores of athlete's foot, surgical wounds, corneal wounds, and hemorrhoids : 1 $, remedy for wounds including pressure sores etc.) Alveolar pyorrhea-Includes so-called hair fever, such as anti-inflammatory agents such as millet and sunburn, hair restorer and hair restorer o

For cosmetics, apply to the skin, for example, after shaving D, after using a depilatory cream, after using a detergent, etc.)), or apply to skin with rough skin, for example. It has excellent skin protection and improvement effects, and Sensitive, E mode Li E down shea one against KawaIsao while恃与a smooth feeling ^like: Application Benefits Conclusions zone preparative Ko杲, affinity etc. remarkable and tool - can be improved 'and also - stimulation to KawaIsao Cosmetic water V Cosmetics, liquids, vans, skin cosmetics in the form of dae-yeon, etc. and, for example, Chan 7 °, skin, hair liquid, Scalp, hair scalp and other scalp; various forms of hair and cosmetics applied to sashimi

The agent for treating wounds, hair follicles and cosmetics of the present invention, which is required to contain as an active ingredient, it is essential that it contains McLogbrin, and Macuglin is egg white. Is known as a glycoprotein poverty that exists in the United States.] The method of preparing the services is already known. [Finy (Feen.yB-E.) Et al. Biochemistry-And 'Fisher, 1st Corap.Bi 0 chem.Pliy »i» 1.), 5' 4r A, 28 1 1 9 7 6) J. Bia-chem., 92, 1679-1682 (1992), 93 , 1 2 1 to 1 2 7 (1 9 8: 3) _β and Nagase et al., Si, Op. ), Vol. 2 58, 1 12, 74 β 1 to 7 48 θ (198 3).

The use of this omega π-globulin as an additive for a cell culture medium has already been known (if it is refreshed, Japanese Patent Application Laid-Open No. 60-23979 S9). Egg white, which is a raw material for preparing the above-mentioned Ovomak mouth globulin, is not particularly limited, and various animal's can be used. Egg whites such as vegetative, quail, turkey, etc. are preferred.Oboma globulin preparation methods from egg whites are also not particularly limited, and various types of proteins generally used for separating protein components are used. According to the method, various operations utilizing the physicochemical properties of Oboma Gloglobulin, such as protein precipitant treatment, molecular chromatography (gel filtration), a. on-exchange換Ku Russia Conclusions grayed La off I-1 centrifugal雜, electrophoresis. _¼ dialysis, such as Ru can and either alone or in combination row of cormorants child.

Examples of a method for obtaining ovomacroglobulin from egg white include mixing egg white with a water-soluble solvent such as a tris-hydrochloric acid buffer solution, or mixing the egg white with a water-soluble solvent. After removing insoluble proteins such as opomucine by adding a gel to the gel, the gel may be filtered. Ovomacroglobulin can be obtained as a glycoprotein ₀

The therapeutic agent of the present invention can be used to prepare a monophasic medical preparation according to a known method, except that the above-obtained glomacrin globulin obtained as described above is blended as an active ingredient. O The preparation can be prepared. O The form of the pharmaceutical preparation can be appropriately selected from various forms according to the intended use of the obtained preparation. Examples of the form include liquid coating agents, lotions, aerosols, and linear agents. G> Ointments >> Injectables such as pills ¾ In addition to external preparations, suppositories >> Examples of preparations for local administration such as injections etc.0 Usually used for the preparation of these various forms Diluents 1) Excipients and the like can be used as appropriate.o For example, in the preparation of an ointment as an external preparation, an ordinary hydrophobic or hydrophilic base such as fat, Fatty oils, lanolin, decelin, raffin, roux '' Glycols, high-grade alcohols, glycerin, water, etc. can be used. In addition, the above-mentioned external preparations may contain various additives which are known to be usually added as needed, for example, stabilizers, fragrances, 1 colorants, etc. 0

The amount of the active ingredient to be contained in the therapeutic agent of the present invention, i.e., the amount of Biamacroglobulin is not particularly limited and may be appropriately selected from a wide range.

30% by weight o

Also dosages and methods of the present invention the therapeutic agent in the form status of _¾ formulation, the active ingredient content in the formulation, determined this age I sex and other conditions of the patient to be applied, by Ji fS on the degree of diseases and the like For example, a therapeutic agent in the form of an external preparation can be applied to the affected area one or more times a day by spraying, applying, or the like in an amount sufficient to reach the entire affected area. it can.

Especially in the case of hair fly medicine> For example, it can be applied to hair and scalp 0

When used as a cosmetics department, it is the same as ordinary cosmetics except that Ovomaclog Mouth Brine is contained as an active ingredient. Thus, various forms as described above, for example, lotion as a cosmetic applied to the skin, various forms such as cream 1 liquid solution _foundation, and for example, Chambers / Rinses * Hair liquids, set-downs, etc. Various forms applicable to scalp or hair such as tonic o The preparation into these various forms can be carried out according to a conventional method. >> At that time, various known cosmetic bases and, if necessary, various incense, antioxidants, surfactants The same applies to the use of additives such as preservatives.

The amount of ovomacroglobulin in the cosmetic can be appropriately selected depending on the form of the obtained cosmetic, the desired effect, and the like. 0 1 to 30% by weight >> Preferably, the amount should be in the range of about α ο ο ο ι to about 1% by weight ο

Hereinafter, the present invention will be described in more detail with reference to Reference Examples and Examples.

Reference example 1

Preparation of OvoMacroglobulin:

Egg white 2 O is suspended in a 1 O m Tris-HCl buffer (PH77) containing an equal amount of L% NaC, and added to a leaching alcohol (molecular weight: = 850, manufactured by Tokyo-Danisei Co., Ltd.) to a concentration of 25%, and subjected to continuous kneading and centrifugation (10,000 rpm) to collect the supernatant. Add the same polyethylene glycol to the supernatant obtained above. Add the solution to a concentration of 1 o% and repeat the centrifugation

(L OOOO rpm) was ₀ collected the precipitated »portion was dissolved in the above瑷銜solution, further Centrifugal雜(l OOOO r _Pm, l O min) to the supernatants were collected, Joseph this Column of CLOS-6B (manufactured by Pharmacia)

9 O O mm) and eluted with the same buffer at a flow rate of 316 Z hours.

For the elution category, Tri using casein as a substrate: 7 ° cin inhibition activity was determined by the method of Kitamoto et al. [T. Kitam. to, Μ.

Naka shima, and A. Ikai, J. Biochem., 92, 1679-

1 682 (1982)) and collected the fractions with trypsin inhibitory activity o ''

The active fraction obtained in the next step was reduced using a Pelicon cassette (MilliAria) equipped with a molecular sieve membrane l OOO OO, and a 5 m -The buffer was exchanged with hydrochloric acid buffer solution (pH 77). O The sample obtained was replaced with a 1 Om tris-salt buffer solution (pH 7) containing 1 OmM NaCZ. 7) DEAE triacyl M equilibrated in

(risacryl, manufactured by LBK) 0 column attached to the column (size 50Χ80 mm) ₀ Wash the column with the same buffer ¾ After washing thoroughly, 1 OmM containing 5 OmM NaCZ Tris-HCl buffer (pH 7.7) Contains 675 «^ and 150! 11¾1 NaC

Elution was performed with 107.5 mM Tris-HCl buffer (pH 7: 7) 675 ^ for 25 hours each, for a total of 5 hours. Under these conditions, the trypsin inhibitory activity category is 70 mM One 1 O—

O eluted between 1 2 O m

The above trypsin inhibitory fractions were collected and dialyzed against 1 = M phosphoric acid 缓 mouth liquid (PH74). O After sufficient dialysis, the dialysis solution was freeze-dried (Raboko Olyophilized by

According to the above >> A single purified Ovomacroglobulin purified sample 試料 9-71 was obtained o

Purification sample Nitsu Te * 4 N meta emission scan Le Ho phosphate, IIOV ₄ after 2 4 hours hydrolysis (in dark圧封tube), A Mi Bruno Sana Na La Lee The one (8 3 5 - 5 Type 0, Hitachi High-Speed Amino Acid Analyzer, manufactured by Hitachi, Ltd.) ) Analyzed o

The result is shown in Table 1 below)) o

First

Amino acid content (mol%)

A s 1 Q3

T h r

S e r ao

G 1 u 1 L6

P ro 4.3

G 1 5.1

A 1 5.8

C y 3/2 1.8

V a 1 8.2

Me t 2LO

I 1 e β5 T ry a9

P h β 4,8

4,6

H s L8

A r g 36

Example 1

Inhibition test of vascular permeability by burns:

In this test, a rat skin is burned with an electric iron, and then a dye (Evans Blue) is applied to the rat, and the leaked dye in the bodily fluid that exudes to the burn site To determine the amount! ) The effect of the therapeutic agent of the present invention on the treatment of wounds was examined using the vascular permeability of the test substance as an index.

Test is 0, ie, I j? Was carried out in the following way, body weight 2 0 0 to 2 5 0 of the window office Turn-male rats ₀ each group La, which were divided into five or we made two groups each group After shaving the hair of the target site along the midline on the back of each of the kits with a parican, one of them was cut into a circular shape with a diameter of 1 by an electric iron (scale temperature 100 to 110). (° C) for 20 seconds to create a burn site. The other was left as a non-burn site o

Immediately after the burn and 24 hours after the burn was made, saline was injected intradermally into each of the burn areas and the non-burn areas, and the rats in each group were treated with saline (control group). O Each rat in the other group has 1

A similar amount of ovomacroglobulin solution was intradermally administered to obtain the ovomacrog α-brin administration group (experimental group). O

235 hours after each of the above administrations, Q 5% Evans' Blue dye was intravenously administered to the rats in each group.30 minutes after that, that is, 48 hours after the burn was made, The rat was exsanguinated and killed, and the skin at the burn site and the non-burn site was removed, and the fat adhered was removed.

The KawaIsao of l ea ² square, 1 N - to immersion in K0H solution la ^, 3 7 C at 2 0 hours left Q 6 with o Next dissolved in N - H ₃ P0 _4: A cell tons (5: 1 3) mixture was added, after stirring mixer one, rows 2 5 min centrifugal雜in 3 OOO rpm, and the upper潸¾ collected ₀

The absorbance of the obtained supernatant was measured at a wavelength of 62 nm by a spectrophotometer.] Measured o Measured absorbance by the standard curve prepared beforehand. O The amount of blue was calculated.o This value was shown as the average soil standard error.o

The results are shown in Table 2 below.

Table 2

vfffc ^ J ¹ ¾ na 3 $

W 5 ^ w m. W IK ^ group

5? A

i m £ Z site) 0.2 Q2

, ./'Ft* '

Burn area 28L7 soil & 49 16L2 soil; ¾82

Non-burn area 36 soil 0.41 44 soil CL40

Burn area 1

Non-burn area 2 & 0 Sat 5ι32 11.8 Sat 3L55

See Table 2 above! ? (Burn site) - et bus emissions scan * Blu leakage enumeration value (non-burn site), control group 2 SO earth in (physiological saline-administered group) and 3 2 and which was to pair with _¼ experimental group In the (Obamacroglobulin administration group), 1 L8 soil ass! ? , Oho, by administration of macroglobulin》 It can be seen that vascular permeability is significantly suppressed o

Example 2

Skin inflammation recovery test by hair removal cream:

For this test, a total of 2 O male Balb / c mice weighing 25-3O9 were used.

Hair on the back of the mouse to be tested ¾ Clean the hair with a parical b b Hair removal cream (Maghair Hair Mover, manufactured by Kaneu Corporation) Q5 evenly It was applied. After leaving for 30 minutes, wipe off the applied cream with warm water! ) A skin inflammation model was created.

The above model mice were divided into 5 groups of ⁴ animals per group. The following four groups of mice were subjected to a total of four times each of the following softener samples on the day when the skin inflammation was created, and on the next day _k3 and 6

0L 3f was applied to make experimental groups 1-4

Experimental group 1: Hydrophilic ointment of Japanese Pharmacopoeia (Yoshida Pharmaceutical Co., Ltd.)

Cloth

Experimental group 2 = Obamacro log ο brin α ο ι%

Softening application with

Experimental group 3: Omac 'π-globulin CL O O 5

% Ointment applied

Experimental group 4: Oboma Crop Globulin 上記. Ο Ο 1

% Ointment applied

In addition, the remaining one group was treated as a control group without any treatment.

After the preparation of each experimental group (intraspinal application) (including the control group), the skin condition of the mouse of each group was observed macroscopically and histologically every day. It was performed under a microscope after xylin-jodine staining (HE staining).

As a result, in the control group on the first day of hair loss cream treatment, inflammation occurs in the epidermis and the dermis layer, and inflammatory cells such as histiocytes and neutrophils converge, particularly in the epidermis and the dermis following it. Inflammation was observed, and the substance strongly stained by aeon spread.o On the ^fourth day, the dermal image of the epidermis and the dermis following it replaced the inflamed image. ] ?, the inflammation had spread throughout the hair follicles

Experimental group 1 (hydrophilic ointment applied group) There was no difference between ₀

On the other hand, the experimental groups 2 to 4 (the group to which the ointment containing oomacroglobulin was applied) had better control of the epidermis than the control group and the experimental group 1 because the inflammation of the dermis and hair follicles was well suppressed. D> The dermal image was also found to be well recovered. Incidentally, the effect seemed to be slightly weaker in the group in which the amount of Oboma clobulin was low (Experimental group 4).

In the experimental group 3 (Ovomacroglobulin 0105% ointment-added ointment group), the results of microscopic observation on the 8th day of hair removal cream treatment]), on the 8th day, a set緣像be clearly compared to day 4 above, new hair follicles subcutaneously KumiSatoshi portion that Ru grown was confirmed (see Fig. 2) ₀ this. trend, the experimental group 1 (Ovo's macroglobulin-free hydrophilic soft spine application group) was almost the same (see Fig. 3).

In addition, although the normal shape of the skin portion of the normal mouse is irregular and wavy, in the experimental groups 2 to 4 of the present invention, the repair of the inflamed surface progresses, and Beginning with the uneven wavy shape] ?, and the breeding and growth of hair groups related to hair growth is remarkable! ) Along with that, hair regrowth was recognized earlier (for example, see Fig. 2).

Implementation Kiyoshi 3

Burn recovery test 2

For this test, Ba lb c males weighing 25 to 30 f 5 groups of ⁴ mice per group were used o

The hair on the back of the mouse under test was cut off with a pallicum.] »Take» Apply hair removal cream and leave it for 5 minutes (to cause inflammation) >> O with warm water

Then:

An electric iron set at 400 (»-27» paraffin cutting / melting iron * made by Takashima Shoten) was applied for about 5 seconds to burn.

The next day after burn load]? Every 3 days, each of the following test softenings was applied to each of the _k burns in an amount of 0.2 to each, and the weight variation of the test animals was observed.

Experimental group 1: Japanese Pharmacopoeia hydrophilic ointment (Yoshida Pharmaceutical Co., Ltd.)

Experimental group 2: Ovomacroprin α ο ι%

Soft blue coating with

Experimental group 3: Ointment containing 0.005% of Obomacroglobulin added to the above cartilage

Experimental group 4 = Oboma Kuro Groslin α ο ο 1

% Softening application

One group of [Remaining]? Was treated as a control group without any treatment.

The results of the weight change measurement in the above test are shown in Fig. Ο.The figure shows the results of each group daily up to 4 days, with the day when the burn was caused by the electric iron as “Day 0”. Animal body The weight change ratio (the average weight before burn injury is set to “1” and the weight change instruction for this) is displayed. The horizontal axis indicates the number of days (days) and the vertical glaze indicates the weight change ratio. Middle (Experimental group 4 (Ointment coated with o-macroglobulin α ο ο! ■%); (2) Experimental group 1 (Ovomacroglobin without π-brin added hydrophilicity) Ointment applied group) and (3) indicate control group ₀

The above weight change reverses the degree of the burn well. That is, after the burn is injured, the vascular permeability is increased due to local inflammation. ? Edema, and therefore, the drowsiness increases according to the size of the burn area, and the weight increases ο

Fig. 1 showing the results of weight change! ), And in the OVOMACLOG π-brine coated group (Experimental group 4) of the present invention, the uncoated control group and the hydrophilic ointment coated group without the OVOMACLOG mouth Brine (Experimental group 1) were used.盧抑え浮浮め判判め.

According to the results of the macroscopic observation in the above test >> 6 days after the injury >> Compared to the control group and the experimental group 1, all of the experimental groups 2 to 4 of the present invention showed very good recovery. Is clearly evident from the comparison of Fig. 4 (photograph of the affected area on day 6 of the injury in experimental group 4) and Fig. 5 (same photo of the control group).

Furthermore, histological observation by HE staining in the above test was performed in the same manner as in Test Example 2.o Results> 4 days after burn injury * In the control group and the experimental group 1, the epidermis * dermis part was completely replaced with a burn-specific eosin and dyeable substance ^ * The morphology remained (see Fig. 7 which is a microscopic photograph of the control group 4 days after the burn injury). In contrast, in the group to which the therapeutic agent of the present invention is applied (for example, experimental group 3), the repair of the dermis starts from the portion in contact with the subcutaneous tissue, and the dermis-like tissue has begun to form. ! ) It was confirmed that hair root formation was slightly different (Experimental group 3 4 days after burn injury, ie, soft spine-applied group to which 5% of Bomacroglobulin 0iOO was added). (See Fig. 6, which is a microscopic photograph of).

From the above results, D> According to the application of the wound healing agent of the present invention, the effect of preventing edema after the burn and the effect of recovering the damaged part were confirmed visually.緣 Formation promoting effect) was clearly recognized.

Example 4

Sensory test of the cosmetic of the present invention

Rough a select feeling Jill female 1 O name (2 ^5-3 5 years old), the cosmetics of the present invention shown in the following Formulation Example 1 (scan key down Mi Torque), 2 times after getting up and before going to bed every day, Two weeks after the skin was applied to the skin, the following three items were evaluated for the skin to which the test cosmetic was applied.o

Also, as a reference test, another control cosmetic sample was prepared in the same manner except that oho * macroglobulin was not used in place of the above cosmetic sample of the present invention. Rough skin Panelists of 5 women (tuna in the same year) who feel the feeling> Conducted a sensory test o

In Table ₀ below, the results of the sensory test are shown in Table 3 below, where the numerical value (numerator Z denominator) indicates (one paneler who responded as good or good for each evaluation item). Inventive cosmetics Control cosmetics

Gently! Feeling 9/10 1/5

Smooth skin feeling 10/10 2/5

Eliminating rough skin Q / 1 O 2/5

See Table 3 above! ? However, the cosmetic preparation of the present invention was proved to be excellent in a _four- sensory test due to the formulation of Ovomacroglobulin.

Example 5

Skin recovery test:

(1) the rabbit back, shea Luba over Na Lee off at 2 5 mm X 2 5 mm size, this time the depth L 2 mm min layer KawaHiroshi deficient wound to create 5 Chikarasho ₀ of each wound Osamu薬剤 In order to eliminate the shadow hairpin of the quiz, the following drugs (~ (D)) were applied once a day as the ₀ drugs to be created with an interval of 3 Omm or more between each defective wound. After the application, cover the area with gauze (25 mm Χ 25 ιηπι large) and apply a breathable nylon film to avoid contamination. (2) Tegaderm> made by Sleam Co., Ltd.) * Close the whole with elastic wrapping. O This procedure was carried out by relaying during the observation period. (Drugs)

(A) Hydrophilic serine ointment

Sarasimi Roo 8

Steal Real Call 3 9

Corel mouth 3 9

White Celine 86 f

7 ° mouth Pilpara Pen QO 625

(1 O O ^)

(B) Hydrophilic petrolatum containing 1% ovomacroglobulin

5 <½Ovomacroglobulin aqueous solution 2 Of

(Contains 4θ2 ^^ methyl-parapene) '-Hydrophilic serine base 80

(c) Hydrophilic petrolatum containing α 1% omacroglobulin

α 5% Ovomacroglobulin aqueous solution 20

(Contains 4 Offl ^ methyl parapen)

Hydrophilic petrolatum base 80

(D) O.O Hydrophilic ボ -serine ointment containing 1% ovomacroglobulin

0.05% Ovomacroglobulin aqueous solution 20 f

(Contains 4 O-methyl parapen)

Hydrophilic serine base S Off

(2) Evaluation of So偬healing, immediately after wounding, and 7 _¾ to 1 4 2 1 day, were observed and related to the following: o (Ii) Wound epithelialization

Photographing and evaluating the wound healing process at each time

(Mouth) Organizational observation

'Biopsy of wound surface at each time, HE stained specimens were used to observe collagen morphology and morphology and epithelium formed in granulation tissue o

No side effects

Observed irritation to normal skin (redness) and presence or absence of dermatitis o

(Result)

The size of the wound on the 7th day after applying the ointment containing omac c globulin αα ι% and α ι% respectively to the experimental skin (See Figures lo and), which was slightly reduced (see Figures 8 and 9), and the formation of granulation tissue was observed on histological findings. The wound healing effect of Ovomacroglobin is further enhanced, and the size of the wound is α ο 1% »ο. 1% The ointment-containing ointment group was clearly reduced ο and the difference was also clear in the agglutination. »There was a difference in the formation of granulation tissue, collagen content, epithelial tissue growth, etc. Admitted. On the 21st day of use, compared to the hydrophilic ointment group (see Fig. 13) on the 1st day, the α-o 1% and α 1% -year-old * softening application group containing macroglobulin (1st day) See Figures 1 and 12). Ο Short-term wound healing period ο This effect was anatomical, promoted granulation, increased collagen protein content, and promoted clotting of epithelial cells ο in addition, it was observed Obomaku log Russia Breakfast Li down軟資was applied _k its side effects against normal skin "was not observed the onset of normal skin redness also rather ¾, or汔dermatitis ₀

In addition, the group applied with 10% oho * gloglobulin-containing ointment showed the same effect as the group applied with αι% oomaguroglobulin-containing ointment.

Example 6

Stretching test for corneal epithelial cells 2

2.5 ~ ao 'rabbits, Pento Norbital

Intravenous administration of 30 ^ Zk? (Pittman Moore)]? Anesthesia performed o The cornea was then removed and a 2x4mm strip ^: corneal piece was prepared did.

Oboma cloglobulin 一 TC 05

The corneal pieces obtained on * were cultured for 28 hours in a culture solution dissolved at U.f / mi »or 0t 5iZm £, 5μ? ZW at each temperature. After culturing, the tissue was fixed with 5% glacial acetic acid (95% ethanol), embedded in raffin, and 4 ^ sections were prepared.o This was subjected to normal HE staining Then, the length of the corneal epithelial cells that had been observed and extended under a microscope was measured.

As a result, as shown in Fig. 8, Ovomacroglobulin promoted the extension of corneal epithelial cells in a volume-dependent manner. O In the figure, A represents the control group containing only TC-199 culture solution. Show, B is the same as the above culture solution. Macroglobulin 0.05 ZΛί, C is Q5? / n and D include 5 # / «^, respectively. Implementation Kiyoshi 7

Periodontal fiber wound healing test *

Wounds were formed in the rat periodontal tissue by the following methods 1 and 2, and the therapeutic effect of the therapeutic agent of the present invention was examined.

(Wound formation)

1. Using a 12-week-old rat, insert a hooked probe near the central part of the upper and lower second white teeth of the sulcus, and reciprocate along the crown several times when reaching the bone. Then, the wound is formed. 0 After that, insert K-File No. 40 for root canal treatment right below the contact point of the left second molar, and move between the left side and the right side. Then, wrap the silk thread (blade silk) 4-O once around the second molar, and put the knot on the side o. After confirming hemostasis, apply the paste o above All manipulations were performed under ketalal intraperitoneal anesthesia.o

2. Gingectomy »Incision is made from the distal molar of the third molar to the mesial of the first molar with the replacement blade of # 12 bird 'beak (bird beak). Put it in the same way and reciprocate to remove the gums.]? The 0 bone surface was exposed to about α5 mm o

(Test paste)

(A) Control paste

Hydrophilic petrolatum base

(B) Paste containing Q05% Obo macroglobulin CIO 25% o ^ macroglobulin: d solution 20 f

(Contains 40 / ^ methyl paraffin)

Hydrophilic petrolatum base * aof

(c) Paste containing αι% pomacroglobulin

Q5% OH * Macroglobulin aqueous solution 2 O ^

(Contains 4 OB ^ methyl rapene)

Hydrophilic serine base 80?

* Hydrophilic petrolatum base (equivalent to l O O?)

Sarasimi Tsuru 8?

Stearyl alcohol 39

Restrol 39

White petrolatum '86?

7 ° mouth pill 9 rappen Q0625 ^

(Administration method) ''

Each paste is put on a rattle once a day

1 is applied 5 minutes ο coating amount is average once about Ο. 3 2 Der Ru ο after application, 2 hours was _k or without giving欽水, first-time coating line cormorant o the above operation after confirmation hemostasis was carried out by intraperitoneal anesthesia of all the values down porcine Lumpur (α ι) ₀

( Evaluation method )

Evaluation method, in 1 »2 ₄ 7 days after the start of the experiment ^置Ra Tsu door were sacrificed, retrieve the lesion! ? Breakfast A down fixed, the door re-click π mouth acetic acid demineralization, after La full fin embedded, by the HE staining _¼ or § The emissions staining]? »Inflammatory cell number, a change in the co-error gain down锇維Observed. (Result)

On the first day after application of the external preparation of the present invention, the effects of wound treatment on inflammatory cell infiltration, hemorrhagic dysfunction in tissues, and collagen collagen condition were as follows. ! ? O

① Inflammatory cell infiltration and tissue ② Internal bleeding

In the control group, there were dilatation and hemorrhage of strong capillaries from the gingival sulcus epithelium to the alveolar lamina from the alveolar bone top. >> The _fibrin layer and scattered neutrophils were infiltrated around it. and (a 1 beta Figure) ₀ - how, the present invention external preparation applied group, lamina propria _¼ bleeding is rather Do strongly despite expansion of capillaries are in the lamina propria are already off Lee Bed Li down layer to be crack 'was on and Ru trend _0. also off Lee Bed Li down layer surface of the invasion is only had less o gingival-groove peel "junctional epithelium of neutrophils in the chromatic) 1 seen in the control group However, localized and intense neutrophil infiltration was observed. α ο ο 5% Ovomacroglobulin paste-applied group and αι% Ovomacrog mouth paste-containing paste group There was a tendency that the dilatation of the capillaries in the lamina propria and the bleeding image were smaller, and that the neutrophil infiltration in the epithelium was less. Fig. 15 is a photograph showing the cross section of the structure of the group coated with 0.05% Obomaclog π-purine-containing paste.

② Collagen fiber condition

Collagen fibers were changed by azane staining ο

In HE staining, the control group is located near and around the wound. One 2β—

The collagen fibers around the wound were ruptured, degenerated, etc., whereas the collagen applied near the wound in the group to which the external preparation of the present invention was applied was torn, but the tears were around the wound. Ο α ι% Ovo macroglobulin-containing paste is more likely to spread α α O 5 o / ₀ Ovo macrolog mouth Brine-containing paper The tendency was stronger than that of the group with the applied ο.

Example 8

Hereinafter, examples of preparation and formulation of the preparation of the present invention are shown.

Formulation Example 1: Cosmetic Formulation of the Present Invention

Preparation of skin milk ''

Flow No. 9 Raffin (100-1 lOc ρ) 7 parts

Cetyl alcohol 015 parts

8 5% Glycerin 7 parts

Sai Bomacro Glo Purin Ao Part 1

7 * Mouth 7 ° mouth Bill Lanolin α ο 3 parts

Acidamide sulfate preservative (methyl parapen) 0.2 part

• Fee i

Add water to make the whole amount loo.

Based on the above composition, o-macroglobulin, zomethylamino: 7 ° opening pyrranolinamide, salfate and glycerine After mixing the components, water was added at 75 ° C under stirring to prepare a homogeneous mixture. Off fin, in degrees Chi Le A Le co Lumpur and main switch le Roh 9 Paix down the thereto while maintaining the mixed under挽拌under "75 heated to prepare a uniform mixture _k same temperature The prepared homogeneous mixed solution was gradually added and stirred and mixed, and allowed to cool to room temperature. Finally, a perfume was stirred and mixed with the prepared solution, and the skin milk of the present invention was obtained. Got o

Formulation Example 2: Preparation of Skin Cream of Cosmetic Formulation of the Present Invention

Fluidized raffin (100 ~: L10Cp) 5 parts

Iso ° Pirmi Restate l O part Stearic acid 3 parts

Cetanol 2 parts

85% glycerin l O part

Lioxydiethylene (E0 = 4) stearate 1 copy

* Macroglobulin αοοι part * Methylamino 7 ° mouth pyrranolic acid 0.04 part

Amy doshi * sulfate preservative (methyl rapene) α2 part

Perfume Μ Add heavy water to make 1 ο ο part ο

₀ was prepared present invention scan rk click Li one arm in the same manner as in Formulation Example Te based on the above composition

Formulation 3: Cosmetic formulation of the present invention

Preparation of skin lotion

95% ethyl alcohol 20 parts 8 5% Glycerin 5 parts Lithium xylene (E 0 = 2O) sol as part

Vitanlauric acid monoester Ovomacroglobulin aoo 5 parts Gumethylamino mouth t? Lulanolic acid ama ao 4 parts

ヅ D-Sulfate Preservative (Methyl Lapene) C12 part

Flavors and dyes qs Add water to make the total amount 1 O O

Based on the above composition, first, ovamacroglobulin, methylaminopropyl lanolinic acid amide, sulfate, and glycerin are referred to. After mixing well, I prepared a homogeneous mixture by adding water. O—After dissolving the fragrances, dyes and preservatives in ethyl alcohol >> ^ Roxyxylene A uniform mixture was prepared by adding monoester of rubitan-lauric acid, and the homogeneous mixture prepared above was added thereto under vigorous stirring and passed. Thus, the skin lotion of the present invention was obtained.o

Formulation Example 4: Cosmetic formula of the present invention

Preparation of skin mouth

A skin lotion of the present invention was obtained in the same manner as in Formulation Example 3 except that the amount of globulin added was changed to αοοοοι part. ₀

Formulation Example 5: Formulation of the wound treatment agent of the present invention

Abo 9 Preservative qs

Appropriate amount of flavor

Add distilled water to make the total amount l O O

Distilled water was added to the above-mentioned ovomacroglobulin, preservative and fragrance to make the total amount lOOW, and then sterilized to prepare a wound treatment agent of the present invention in the form of a solution for use in olive oil.

Prescription Kiyoshi 6: Formulation of the wound treatment agent of the present invention

Ovomacroglobulin αθ5 ^ Preservative

Appropriate amount of flavor

Add distilled water to make 1 量全 ο

In the same manner as in Formulation Example 5, a wound treatment agent of the present invention in the form of a spray solution having the above composition was prepared.

Formulation Example 7: Formulation of the wound treatment agent of the present invention '' Preparation of hydrophilic cartilage

O'Macro Groplin Q5?

White cellulose 25 O

Stearel alcohol 22 Of

7 ° lobylene glycol 12 Of sodium sodium lauryl sulfate 15?

Ethyl oxybenzoate or Q25? Methyl benzoyl benzoate La xy benzoate: 7 ° mouth pill a L 5 ^

purified water

Total volume 1 ο ο ο ^ One 3 O—

The above ingredients were blended to prepare the hydrophilic ointment-type wound treatment of the present invention containing Oboma clobulin o Formulation Examples S to 11: Formulation of the therapeutic agent useful for the present invention

Preparation of parenteral ointment

Formulation Example 7> In the same manner, the amount of ovomacroglobulin was changed to O.01, 0.05, O.1? Prepare therapeutic agent o

Industrial applicability

Thus, when the obtained therapeutic agent of the present invention is used as a wound therapeutic agent, it has an excellent effect on promoting wound healing as described above. Demonstrate excellent results o

In addition, when the therapeutic agent of the present invention is used as a hair restorer or a hair restorer, it has an effect of promoting hair growth, and also has an effect of protecting and improving hair, for example, eliminating roughness of hair and improving gloss and the like. O

Furthermore, when used as cosmetics, it can exert an excellent protective and improving effect on the skin, and can be used in place of conventional cosmetics of this type.)) It is also advantageous. The therapeutic agents and cosmetics of the present invention are also excellent in their storage stability and safety.

Claims

The scope of the claims

W. A therapeutic agent selected from wound treatments and hair flies containing J. ovomacroglobin as an active ingredient o

治療 The therapeutic agent according to claim 1, which is a wound therapeutic agent.

3. Wound sores, cuts, burns, burns, frostbite, skin m, dry skin keratosis, cracks, redness, skin] »Flame * Athlete's foot sore, surgical wound, corneal wound _fc hemorrhoid And the wound treatment agent according to claim 2, which is selected from a pressure sore o

4. The wound treatment agent according to claim 2, wherein the wound is burned or heated o

5. The wound treatment agent according to claim 2, wherein the wound is a membrane wound o

6. The wound treatment agent according to claim 2 which is an inflammation treatment agent.

7. The wound treatment agent according to claim 5, which is a treatment agent for alveolar pyorrhea o

8. The therapeutic agent according to claim 1, which is a hair follicle such as a hair restorer or a hair restorer.

9. Obamacroglobulin ¾ α ο ο ο ι 〜 3 ο Weight ⁰ / o The therapeutic agent according to claim 1 containing ο ⁰ / o

10. Cosmetics containing omacroglobulin as an active ingredient o

11. The cosmetic according to claim 10, which is a skin cosmetic.

12. The cosmetic according to claim 11, wherein the form is any one of a lotion _k cream, a milky lotion, and a foundation.

13. Cosmetic according to claim 10 which is a cosmetic for hair o

14. Forming force: 7 ° — Claim 13, which is the difference between the lens, hair liquid, settling and hair tonic. Listed cosmetics ₀

15. Cosmetic according to claim 1, comprising α macroglobulin α αοοο 1 to 011% by weight.

16. A method for treating wounds characterized by applying oho macroglobulin to the affected area ο

17. A hair nourishing or hair-growing method characterized by applying Ovomacro globulin to hair or scalp.

18. A skin protection method characterized by applying Ovomacroglobulin to the skin.

19. Use of OvoMacro Glin for the manufacture of therapeutic agents ο

20. Use according to claim 19, wherein the therapeutic agent is a wound therapeutic agent ο

21. Use according to claim 19, wherein the therapeutic agent is an inflammation therapeutic agent.

22. Use according to claim 19, wherein the therapeutic agent is a hair follicle such as a hair restorer or a hair restorer ο

23. Use of Oboma Kurogloblin for the manufacture of cosmetics -33-for o