JP2002226395A - Agent for promoting cure of skin wound - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a wound healing agent for skin wound such as laceration, scratch, surgical incision, skin ulcer and scald. SOLUTION: A remarkable cure promoting action on skin wound is developed by an insulin-like growth factor I in combination with Arg-Pro-Lys-Pro-Gln-Gln- Phe-Phe-Gly-Leu-Met-NH2 or Phe-Gly-Leu-Met-NH2. Accordingly, the promoting effect on the growth of the epidermis and on the heal of skin wound can be developed by the administration of an insulin-like growth factor in combination with a substance P analog and/or its pharmacologically permissible salt.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a skin wound healing promoter comprising at least one substance P analog and a pharmaceutically acceptable salt thereof and an insulin-like growth factor as active ingredients.

[0002]

BACKGROUND OF THE INVENTION Skin wounds are damages to surface tissues such as lacerations, abrasions, surgical incisions, skin ulcers, and burns. The treatment of such skin wounds involves first-aid treatment of the injured site,
It is common to wait for the body to heal naturally by its own resilience.

[0003] However, such natural healing requires a long period of time to recover and pain continues. Therefore, it is necessary to actively promote wound healing by administering a wound healing agent to the wound site. Is desirable.

[0004] In the process of wound healing, new epithelial tissue and connective tissue are formed by the migration and proliferation of cells.
An agent that promotes or stimulates migration, differentiation, and proliferation of cells involved in wound healing can be a wound treatment agent. Lysozyme chloride, solcoserine and the like are known as such wound treatment agents.

[0005] However, existing wound therapeutic agents have a problem in that the effect of promoting wound healing is not sufficient, and the wound cannot be healed in a short period of time. These wound treatments are thought to be due to their small contribution to epidermal recoating, collagen synthesis, improvement of peripheral circulation, granulation, and angiogenesis, which are important factors in the wound healing process. ing.

Meanwhile, substance P analogs are disclosed in US Pat. No. 3,862,114 specifically.
A polypeptide consisting of ~ 12 amino acids,
These polypeptides are known to have an antihypertensive effect. Substance P, one of substance P analogs, is a polypeptide released and produced from nerve endings, and is Arg-Pro-Lys-Pro-Gln-G
ln-Phe-Phe-Gly-Leu-Met-NH
It is a polypeptide consisting of 11 amino acids represented by ₂ , and is known to exhibit vasodilation, smooth muscle contraction, promotion of salivary gland secretion, diuretic action, and the like. In the ophthalmological field, substance P improves the abnormal secretion of conjunctival goblet cells in ocular disorders (International Patent WO95 / 1308).
No. 7 publication) and the dynamics of substance P during inflammation such as keratitis (Journal of the Japanese Ophthalmological Society, 91 , 982-987 (198
7), Journal of the Japanese Ophthalmological Society, 92 , 448-452 (1988)). Furthermore, Phe-Gly-Leu-Met-, which is a tetrapeptide on the C-terminal side of substance P,
It is described in JP-A-10-17489 that NH ₂ (hereinafter referred to as FGLM) has an effect as a therapeutic agent for corneal disorders.

On the other hand, insulin-like growth factor is one of the factors regulating normal human cell growth, such as epidermal growth factor, fibroblast growth factor, platelet-derived growth factor and transforming growth factor. These include insulin-like growth factor-I (hereinafter, IGF-I) and insulin-like growth factor-II (hereinafter, IGF-II). IGF
-I to stimulate thyroid cell proliferation (J. B.
iol. Chem., 264 , 18485-18488 (1989)).
GF-II regulates muscle growth and differentiation (Hu
mol. Gene., 3 , 1117-1121 (1994)).

[0008] However, no action on skin diseases has been reported for any substance P analog or insulin-like growth factor.

[0009]

SUMMARY OF THE INVENTION
It was an interesting task to conduct a pharmacological test on skin epidermis by combining substance P analogs and insulin-like growth factor to examine the effect of promoting skin epidermal elongation and the effect of promoting skin wound healing.

[0010]

Means for Solving the Problems The present inventors carried out the above-mentioned pharmacological tests to obtain substance P or FGLM.
In the presence of a substance P analog represented by (I) and an insulin-like growth factor represented by IGF-I, these agents act synergistically to remarkably promote skin epidermal elongation and skin wound healing. Furthermore, they have also been found to exert an excellent healing effect on intractable diabetic skin ulcers. That is, the skin wound healing promoting agent of the present invention containing at least one of substance P analog and a pharmaceutically acceptable salt thereof and an insulin-like growth factor as active ingredients is useful for producing tears, abrasions, surgical incisions, skin ulcers (diabetic It has a healing promoting effect on skin wounds such as skin ulcers) and burns and other diseases such as gangrene. The skin wound healing promoter of the present invention includes ascorbic acid, ascorbic acid ester, salts of ascorbic acid, pantothenic acid, which have already been recognized as having a wound healing effect.
A salt of pantothenic acid or the like can be added for use.

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION As substance P analogs,
For example, Tyr-Arg-Pro-Lys-Pro-Gl
n-Gln-Phe-Phe-Gly-Leu-Met
_{-NH 2, Arg-Pro-Lys} -Pro-Gln-
Gln-Phe-Phe-Gly-Leu-Met-N
H ₂ (substance P), Arg-Pro-Lys-P
ro-Gln-Gln-Phe-Tyr-Gly-Le
_{u-Met-NH 2, Tyr} -Pro-Gln-Gln
-Phe-Phe-Gly-Gln- Met-NH 2,
Pro-Gln-Gln-Phe-Phe-Gly-L
eu-Met-NH ₂ , Pro-Gln-Gln-Ph
e-Tyr-Gly-Leu-Met-NH ₂ , Gln
-Gln-Phe-Phe-Gly-Leu-Met-
NH ₂ , Asp-Ala-Phe-Tyr-Gly-L
eu-Met-NH ₂ , Gln-Phe-Phe-Gl
y-Leu-Met-NH ₂ , Ala-Phe-Tyr
_{-Gly-Leu-Met-NH 2} , Phe-Phe-
Gly-Leu-Met-NH ₂ , Tyr-Phe-G
ly-Leu-Met-NH ₂ , Gly-Phe-Gl
y-Leu-Met-NH ₂ , Phe-Gly-Leu
_{-Met-NH 2 (FGLM),} Tyr-Gly-Le
u-Met-NH _{2 and the} like. More preferable examples include substance P and FGLM.

The amino acid structures of substance P analogs include L-form, D-form and DL-form, all of which are included in the present invention. A more preferred embodiment of the present invention is a polypeptide having all three-dimensional structures in L-form.

The pharmaceutically acceptable salts of substance P analogs, substance P and FGLM include, for example, hydrochloride, sulfate, phosphate, lactate, maleate, fumarate, oxalate, and the like. Methanesulfonate, p-toluenesulfonate and the like can be mentioned.

[0014] The skin wound healing promoter of the present invention can be formulated using widely used techniques. Ointments, jellies, cataplasms, patches,
Lotions, creams, sprays, aerosols, plasters, suspensions, emulsions, tablets, pills and the like can be exemplified, and can be used as a liquid by selecting an appropriate solvent. In addition, in order to prepare a skin wound healing promoter, fillers, excipients, bases, disintegrants, bulking agents, binders, film agents, lubricants, coloring agents, pH adjustments are used according to the dosage form. Agents, solubilizers, suspending agents, buffers, stabilizers, preservatives, preservatives, surfactants, antioxidants, dispersants, emulsifiers,
Solubilizers, solubilizers and the like can be added.

Examples of the carriers for the above-mentioned preparations include white petrolatum, liquid paraffin, gelled hydrocarbon, cetyl alcohol, polyethylene glycol, gelatin, corn starch, sodium alginate, methylcellulose,
Hydroxyethylcellulose, carboxymethylcellulose, plastic base hydrophilic, gelatin, dextrin, cetyl alcohol, stearyl alcohol, polyethylene glycol, polyvinyl alcohol, methoxyethylene-maleic anhydride copolymer, polyvinyl ether, polymer containing vinylpyrrolidone・ Copolymer, sodium stearate, magnesium stearate, benzalkonium chloride, olive oil,
Fats and oils such as camellia oil and soybean oil, lactose, water and the like.

The amount of the substance P analog and the pharmaceutically acceptable salt thereof contained in the skin wound healing promoter is 0.001 to 10% by weight, more preferably 0.005 to 5% by weight. % Range. Further, the amount of insulin-like growth factor contained in the skin wound healing promoter is 0.0001 to 5% by weight, more preferably 0.001 to 1% by weight.

The skin wound healing promoter of the present invention can be administered in various forms depending on the site of the wound and the degree of the wound. For example, when used as an external preparation, it is desirable to directly apply, spray or apply the present preparation to a required site (affected part) such as the skin.

The dosage of the skin wound healing promoter of the present invention is as follows:
It can be appropriately selected in consideration of symptoms, age, dosage form, and the like. The substance P analogs and pharmaceutically acceptable salts thereof are generally 0.01 to 5000 per day.
mg, preferably 0.1 to 1000 mg, which is administered in combination with (mixed with) insulin-like growth factor in one or several divided doses. In addition, the dose of insulin-like growth factor is usually 0.001 to 1000 mg, preferably 0.01 to 500 mg per day, and the substance P analog and its pharmaceutically acceptable salt are divided once or several times. Is administered in combination (mixed) with at least one of the above. The skin wound healing promoting agent of the present invention is 1 to 1 day.
It can be administered in five divided doses.

In the following, preparation examples and results of pharmacological tests are shown, but these examples are for better understanding of the present invention and do not limit the scope of the present invention.

[0020]

EXAMPLES [Preparation Examples] Representative preparation examples of the present invention are shown below.

Example 1 (Ointment A-1) Substance P 100 mg IGF-I 10 mg Liquid paraffin 10 g White petrolatum suitable amount in 100 g

The concentration of 0.2% (w / w) (ointment A-2), 0.5%
% (W / w) (ointment A-3) and 1.0% (w / w) (ointment A-4) can be prepared.

Example 2 (Ointment B-1) 100 mg of FGLM 100 mg IGF-I 5 mg Liquid paraffin 10 g White petrolatum Appropriate amount

By changing the amount of FGLM added, the concentration became 0.3% (w / w) (ointment B-2), 1% (w / w).
w) (Ointment B-3), 2% (w / w) (Ointment B-4) ointment can be prepared.

Example 3 (Ointment C-1) FGLM 50 mg IGF-I 10 mg Ascorbic acid 3 mg Liquid paraffin 10 g Plasticy base hydrophilic in 100 g

The concentration of 0.3% (w / w) (ointment C-2), 1% (w / w)
w) (Ointment C-3), 3% (w / w) (Ointment C-4) ointment can be prepared.

[Pharmacological test] (1) Effect on skin epidermal elongation After excessive administration of pentobarbital to rats (Spraque-Dawley), ear tissue pieces of about 2 x 4 mm were prepared. 60m
Each test drug [Substance P, F in Table 1]
18 ml of TCM199 medium containing GLM, IGF-I, substance P + IGF-I and FGLM + IGF-I] was added thereto, and three pieces of tissue were added to each dish (12 cases per group: 3 sections). After culturing for 24 hours in a CO ₂ incubator (37 ° C., 5% CO ₂ ), the tissue slices were fixed with ethanol-acetic acid (volume ratio: 95: 5), and three tissue slices per tissue slice according to a conventional method. Was prepared. After staining with hematoxylin-eosin, the length of the epidermal cell layer in which the cut surface was elongated was measured. As a control, a culture obtained by culturing a tissue piece under the same conditions as described above using a culture solution containing no test drug was used. Table 1 shows the results. In addition, the value in a table | surface shows the average value of 12 examples.

[0028]

[Table 1]

(2) Effect on skin wound healing Streptozotocin (70 mg / kg) dissolved in citrate buffer under inhalation of diethyl ether in rats (Wistar)
BW) was injected from the tail vein to prepare diabetic rats (DM Rat) (5 rats per group). Non-diabetic rats (non DM Rat) were prepared by intravenously injecting the same amount of citrate buffer alone into rats (Wistar) (5 rats per group). Four weeks after intravenous injection, the back hairs of all rats were shaved with a hair clipper under diethyl ether inhalation anesthesia and depilated with a depilatory cream. Twenty-four hours later, five layers of full-thickness epidermis and dermis were formed at equal intervals on the back skin using a trepan for skin biopsy having a diameter of 5 mm. It was applied once a day. Immediately before applying the ointment once a day, a photograph of the wound on the back was taken and the area was measured. Non-diabetic rats (nonDM R
Table 2 shows the area measurement results for at), and Table 3 shows the area measurement results for diabetic rats (DM Rat). In addition, the value in each table | surface shows the average value of 5 examples.

[0030]

[Table 2]

[0031]

[Table 3]

[0032]

[0033]

As is evident from Table 1, even when the tissue pieces are cultured using substance P, FGLM and IGF-I alone, the elongation of the skin epidermis is the same as that of the control (no addition). On the other hand, when the tissue pieces were cultured in the presence of substance P or FGLM and IGF-I, the elongation was 2 to 2.5 times that when these agents were used alone (synergistic effect). Is shown. Table 2 shows that the ointment using substance P and IGF-I together promotes the wound healing of the skin more remarkably than when using substance P or IGF-I alone, and Table 3 shows that It shows that it also exerts an excellent effect on diabetic rats (DM Rat). Therefore, when at least one of substance P analogs represented by substance P and FGLM and pharmaceutically acceptable salts thereof and insulin-like growth factor represented by IGF-I are administered in combination (mixed), these are administered. The drugs act synergistically to effectively promote the healing of skin wounds such as lacerations, abrasions, surgical incisions, skin ulcers, especially intractable skin ulcers and burns in diabetic patients.

 ──────────────────────────────────────────────────続 き Continuing from the front page (72) Inventor Katsuhiko Nakata 8916-16 Takayama-cho, Ikoma City, Nara Prefecture Santen Pharmaceutical Co., Ltd. Laboratory F-term (reference) 4C084 AA01 AA02 AA16 BA16 BA18 DB58 MA02 MA13 MA16 MA23 MA28 MA32 MA63 ZA892 ZC542

Claims (8)

[Claims] 1. A skin wound healing promoter comprising a substance P analog or a pharmaceutically acceptable salt thereof and an insulin-like growth factor as active ingredients. 2. 0.001 to 10% by weight of a substance P analog or a pharmaceutically acceptable salt thereof and 0.02% by weight.
A skin wound healing promoter containing 001 to 5% by weight of an insulin-like growth factor as an active ingredient. 3. Arg-Pro-Lys-Pro-Gl
n-Gln-Phe-Phe-Gly-Leu-Met
-NH _2, at least one of Phe-Gly-Leu-Met- NH 2 and salts which are acceptable as their pharmaceutically
A skin wound healing promoter containing Tsutomu insulin-like growth factor-I as an active ingredient. 4. Arg-Pr of 0.001 to 10% by weight
o-Lys-Pro-Gln-Gln-Phe-Phe
_{-Gly-Leu-Met-NH 2} , Phe-Gly-
Leu-Met-NH ₂ and at least one 0.0001 wt% of salts which are acceptable as their pharmaceutically
A skin wound healing promoter containing the insulin-like growth factor-I as an active ingredient. 5. The method of claim 1, wherein the skin wound is a laceration, abrasion, surgical incision,
The skin wound healing promoter according to any one of claims 1 to 4, which is a skin ulcer or a burn. 6. A skin wound healing promoter according to claim 5, wherein the skin ulcer is a diabetic skin ulcer. 7. A skin epidermal elongation promoter comprising a substance P analog or a pharmaceutically acceptable salt thereof and an insulin-like growth factor as active ingredients. 8. 0.001 to 10% by weight of a substance P analog or a pharmaceutically acceptable salt thereof and 0.0
A skin epidermal elongation promoter containing 001 to 5% by weight of an insulin-like growth factor as an active ingredient.