JP2005281186A - Wound healing agent, agent for amelioration, promotion of healing or prevention of injure or the like of tendon or ligament, functional food and medicine - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a useful wound healing agent, an agent for amelioration, promotion of the healing and prevention of injure, or the like, of a tendon or a ligament, an agent for ameliorating or enhancing the tendon or the ligament, an agent for amelioration, promotion of the healing and prevention of injure or rupture of the tendon or the ligament of a racehorse, an agent for ameliorating or enhancing the tendon or the ligament of the racehorse, and functional food and medicine containing the wound healing agent. <P>SOLUTION: The wound healing agent contains a peptide as a degradation product obtained by degrading a collagen or a gelatin by a collagenase and represented by an amino acid sequence of (Gly-X-Y)n (wherein, Gly is a glycine residue; and X and Y are each an arbitrary amino acid residue except the glycine) as an active ingredient. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a wound healing agent, improvement against tendon damage or tear, healing promotion or prevention agent, tendon improvement or strengthening agent, improvement against ligament damage or tear, healing promotion or prevention agent, ligament improvement or strengthening agent, Racehorse flexor tendon inflammation, damage or rupture improvement, healing promotion or prevention agent, racehorse flexion tendon improvement or strengthening agent, racehorse ligament damage or rupture improvement, healing promotion or prevention agent, racehorse ligament It relates to improving or enhancing agents, functional foods and pharmaceuticals.

  Collagen accounts for about 30% of the protein that makes up the body, and is present throughout the skin, blood vessels, viscera and bone tissue. In the skin, 70% of the dermis is made of collagen, and the fascia enclosing individual muscles is also made of collagen. Furthermore, it is known that the main component of the cell matrix that surrounds the outside of the cell body and serves as a scaffold for supporting the cell population is collagen, and from the past, collagen and collagen obtained by heat-denaturing collagen, and their Hydrolysates have been widely used as raw materials for cosmetics and foods, and further as biofunctional materials for pharmaceuticals.

  In recent years, many studies have been conducted on the pharmacological effects of oral intake of collagen proteins and their degradation products. So far, the main effects include osteoblast proliferation promotion action, bone strengthening action (for example, see Patent Documents 1 and 2), which leads to prevention / improvement of osteoporosis, and improvement of the deterioration of biological tissue function with aging. Numerous effects have been clarified, such as the metabolism promoting action of living tissues (see, for example, Patent Documents 3 and 4), the skin metabolism promoting action, and the skin activation action (see, for example, Patent Documents 4 and 5). .

  Furthermore, according to the study by the present inventors, a collagen tripeptide having an amino acid sequence of Gly-XY, which is obtained by specifically degrading a collagen component or a gelatin component using a collagenase enzyme, is obtained by using conventional collagen, gelatin, Or it becomes clear that it is digested and absorbed more quickly and efficiently than the hydrolyzate, and exhibits various functions including the above-mentioned functions (see Patent Document 6). Collagen production promoters, functional foods, pharmaceuticals, osteoblast or fibroblast growth promoters, bone strengthening or density improvers, biological tissue metabolism promoters, skin activators, skin or bone additives It is possible to provide agents for preventing or improving functional deterioration due to age, chronic rheumatism, osteoarthritis improving agents, and wound healing promoters such as fracture healing became.

JP-A-9-255588 Japanese Patent Laid-Open No. 11-12192 Japanese Unexamined Patent Publication No. 7-278012 Japanese Unexamined Patent Publication No. 9-67262 JP 2000-201649 JP 2002-255847 JP

  Healing of Achilles tendon rupture and ligament injury that occurs mainly during sports often takes a long time, and many athletes lose their lives as a result of a delay in return. Moreover, the problem of flexor tendinitis that develops at the flexor tendon corresponding to the human Achilles tendon is becoming serious not only in humans but also in racehorses. 1,200 racehorses develop flexor tendinitis annually, severely ill horses are forced to retire, and even if they are expected to recover, the return to race is significantly delayed, but the recurrence rate is high, and tendonitis for racehorses Is considered an incurable disease. Under such circumstances, wound healing agents, functional foods, and pharmaceuticals having an effect of promoting healing of tendon rupture, ligament injury, flexor tendonitis and the like are eagerly desired.

  The present invention has been made to solve such problems, and is a novel and useful wound healing agent, improvement against tendon damage or rupture, healing promotion or prevention agent, tendon improvement or strengthening agent, ligament damage. Or improvement for rupture, healing or prevention agent, ligament improvement or strengthening agent, inflammation of racehorse flexor tendon, improvement for injury or tear, healing promotion or prevention agent, racehorse flexor tendon improvement or strengthening agent, racehorse An object of the present invention is to provide an improvement agent for ligament damage or rupture, an agent for promoting or preventing healing, an agent for improving or reinforcing a ligament of a racehorse, a functional food containing the wound healing agent, and a medicine.

  The present inventors have been able to efficiently digest and absorb a collagen tripeptide having an amino acid sequence of Gly-XY, which is obtained by specifically degrading a collagen component or a gelatin component using a collagenase enzyme, even in a small amount. Promotes collagen production, promotes osteoblast or fibroblast proliferation, strengthens or increases bone density, promotes metabolism of living tissue, activates skin, prevents or improves functional deterioration due to aging of skin or bone, chronic rheumatism, It has been clarified that it has functions such as improving osteoarthritis and promoting healing of fractures (see Patent Document 6). Furthermore, the present inventors have found that the collagen tripeptide also has healing promoting effects such as tendon rupture, ligament damage, flexor tendinitis and the like.

  That is, the wound healing agent according to the present invention is a degradation product obtained by degrading collagen or gelatin with collagenase, and the amino acid sequence is (Gly-XY) n (wherein Gly represents a glycine residue). , X and Y represent any amino acid residue other than glycine, and n represents a positive integer).

The wound healing agent according to the present invention is a degradation product obtained by degrading collagen or gelatin with collagenase, and the amino acid sequence is (Gly-XY) n (wherein Gly represents a glycine residue, X And Y represents any amino acid residue other than glycine, and n represents a positive integer).
In the wound healing agent according to the present invention, the degradation product preferably contains 5% by mass or more of a tripeptide whose amino acid sequence is represented by Gly-XY.

Further, the wound healing agent according to the present invention is a degradation product obtained by degrading collagen or gelatin with collagenase, and has an amino acid sequence of Gly-XY (where Gly represents a glycine residue, X and Y represents an arbitrary amino acid residue other than glycine).
The wound healing agent according to the present invention promotes wound healing and contributes to early recovery.

  The wound healing agent according to the present invention preferably contains ascorbic acid or a derivative thereof. In this case, the effect of promoting wound healing can be enhanced.

The improvement, healing promotion or preventive agent for tendon damage or tear according to the present invention is characterized by comprising the aforementioned wound healing agent. The agent for improving, healing promoting or preventing tendon damage or tear according to the present invention has the effect of improving, promoting or preventing healing of tendon damage or tear.
The tendon improving or reinforcing agent according to the present invention is characterized by comprising the aforementioned wound healing agent. The tendon improving or reinforcing agent according to the present invention has an effect of improving or strengthening a tendon.

The agent for improving, healing or preventing ligament damage or rupture according to the present invention is characterized by comprising the aforementioned wound healing agent. The agent for improving, healing or preventing ligament damage or rupture according to the present invention has the effect of improving, promoting or preventing ligament damage or rupture.
The ligament improving or reinforcing agent according to the present invention is characterized by comprising the aforementioned wound healing agent. The ligament improving or reinforcing agent according to the present invention has an effect of improving or reinforcing the ligament.

The agent for improving, accelerating or preventing healing of inflammation, damage or tear of a flexor tendon of a racehorse according to the present invention is characterized by comprising the aforementioned wound healing agent. The agent for improving, healing or preventing the bending, tendon inflammation, damage or tear of the racehorse according to the present invention has the effect of improving, promoting or preventing the inflammation, damage or tear of the flexor tendon of the racehorse.
The agent for improving or reinforcing a flexor tendon of a racehorse according to the present invention comprises the aforementioned wound healing agent. The agent for improving or reinforcing the flexor tendon of the racehorse according to the present invention has the effect of improving or strengthening the flexor tendon of the racehorse.

The agent for improving, healing or preventing a ligament damage or rupture of a racehorse according to the present invention comprises the aforementioned wound healing agent. The improvement, healing promotion or prevention agent for damage or rupture of the ligament of the racehorse according to the present invention has an effect of improving, promoting or preventing the damage or rupture of the ligament of the racehorse.
The ligament improving or reinforcing agent for racehorses according to the present invention is characterized by comprising the aforementioned wound healing agent. The ligament improving or reinforcing agent for a racehorse according to the present invention has an effect of improving or strengthening the ligament of a racehorse.

  That is, an improvement, healing promotion or prevention agent for tendon damage or rupture according to the present invention. An improvement or strengthening agent for tendon according to the present invention. An improvement, healing promotion or prevention agent for ligament damage or rupture according to the present invention. Improvement or reinforcement agent for ligaments according to the invention- Improvement, healing promotion or prevention agent for inflammation, damage or tearing of the tendon of the racehorse according to the invention- Improvement or reinforcement agent for the tendon of the racehorse according to the invention is the aforementioned Wound healing agent ・ Improvement, healing promotion or prevention of damage or rupture of ligament of racehorse according to the present invention ・ Improvement or strengthening agent of racehorse ligament according to the present invention (hereinafter referred to as a tendon improving agent) is collagen Alternatively, a degradation product obtained by degrading gelatin with collagenase, the amino acid sequence of which is (Gly-XY) n (where Gly represents a glycine residue, and X and Y are An active ingredient includes a peptide represented by any amino acid residue other than lysine, and n represents a positive integer. Preferably, the tendon-improving agent is a degradation product obtained by degrading collagen or gelatin with collagenase, and the amino acid sequence is (Gly-XY) n (wherein Gly represents a glycine residue, X And Y represents any amino acid residue other than glycine, and n represents a positive integer). More preferably, in the agent for improving tendon and the like, the degradation product contains 5% by mass or more of a tripeptide whose amino acid sequence is represented by Gly-XY. An agent for improving tendons and the like is a degradation product obtained by degrading collagen or gelatin with collagenase, and has an amino acid sequence of Gly-XY (where Gly represents a glycine residue, and X and Y represent glycine. It is preferable to use a tripeptide represented by (representing any amino acid residue other than) as an active ingredient. The tendon and the like improving agent preferably contains ascorbic acid or a derivative thereof.

  The tendon improving agent may be used for mammals other than humans, racehorses, birds, reptiles, and fish. The target tendons may be Achilles tendons, shoulders, arms, or other parts. The target ligament may be any part. The flexion tendon and ligament of the target racehorse are preferably the flexion tendon and ligament of the leg.

The functional food according to the present invention contains 0.005% by mass or more of the above-described wound healing agent or tendon improving agent.
The pharmaceutical product according to the present invention is characterized by containing 0.005% by mass or more of the aforementioned wound healing agent or tendon improving agent.

  In the present invention, collagenase is derived from bacteria such as Clostridium histolyticum and Bacillus, actinomycetes such as Streptomyces parvulus, lactic acid bacteria, yeasts or fungi, and has a unique amino acid sequence of collagen: (Gly-XY) n on the amino group side of glycine An enzyme that specifically cleaves is used. Collagenase is a gene recombination enzyme obtained by incorporating these enzyme genes into specific vectors by genetic engineering and producing them in other cells or animals such as lactic acid bacteria and yeast, and has similar substrate specificity. It may be a collagenase-like enzyme having

  Collagenase may be used in a free form, or may be used as an immobilized enzyme in which collagenase is bound to various carriers by a physical adsorption method or a chemical binding method. Examples of the enzymatic degradation method using collagenase include (a) a batch method, (b) a column method, and (c) a method combining these. Various combinations of the production line by the methods (a) to (c) and the form of collagenase to be used can be employed.

  The tripeptide has an amino acid sequence of (Gly-Ala-X1), X1 is Ala, Arg, Asp, Hyp, Lys, Ser or Val, (Gly-Asp-X2), X2 is Ala or Thr, (Gly- Gln-X3), X3 is Glu, Hyp or Ser, (Gly-Pro-X4), X4 is Ala, Arg, Hyp, Hyl, Gln, Met, Ile, Lys, Pro, Ser, Thr or Val, (Gly- Ser-X5), X5 is Ala, Glu or Hyp, (Gly-Lys-X6), X6 is Asp, Pro or Ser, (Gly-Leu-Hyp), or (Gly-Val-X7), X7 is Hyp or It is preferably composed of one or a mixture of two or more tripeptides represented by the general formula of Arg. In particular, the tripeptide has an amino acid sequence of (Gly-Ala-Arg), (Gly-Ala-Hyp), (Gly-Ala-Lys), (Gly-Pro-Ala), (Gly-Pro-Arg) , (Gly-Pro-Hyp) and (Gly-Pro-Ser) are preferred to be a mixture of tripeptides represented by the general formulas. The tripeptide may be obtained by a synthesis technique using amino acids or a fermentation technique.

  The wound healing agent, tendon improving agent, functional food and pharmaceutical product according to the present invention may contain either or both of calcium and vitamin D. Further, it may contain glycyrrhizic acid ester or a derivative thereof. Moreover, hyaluronic acid may be included. The wound healing agent, tendon and other improving agents, functional foods and pharmaceuticals according to the present invention may be in the form of a liquid, a solid, a capsule, or the like. Moreover, you may add other additives, such as a sweetening agent, an excipient | filler, a filler, a bulking agent, a surface active agent, a coloring agent, a fragrance | flavor, and an antiseptic | preservative. In addition to oral administration, the wound healing agent, tendon-improving agent and pharmaceutical agent according to the present invention may be administered by methods such as anal administration, intravenous administration, subcutaneous administration, and intramuscular administration. It is preferable to appropriately change the dose according to the age, weight, symptoms, etc. of the administration subject.

  According to the present invention, a novel and useful wound healing agent, improvement against tendon damage or tear, healing promotion or prevention agent, tendon improvement or strengthening agent, improvement against ligament damage or tear, healing promotion or prevention agent, ligament Improvement or strengthening agent, racehorse flexor tendon inflammation, damage or rupture improvement, healing promotion or prevention agent, racehorse flexion tendon improvement or enhancement agent, racehorse ligament damage or rupture improvement, healing promotion or prevention An agent, a racehorse ligament improvement or strengthening agent, a functional food containing the wound healing agent, and a medicine can be provided.

  The wound healing agent according to the embodiment of the present invention is a peptide composition obtained by specifically degrading a collagen component or a gelatin component using a collagenase enzyme, and is a tripeptide (Gly-XY) having an amino acid sequence of Gly-XY The collagen tripeptide) contains 5% or more. In the amino acid sequence Gly-XY, X and Y are any amino acid residue other than glycine (Gly), such as proline, alanine, or hydroxyproline.

  The wound healing agent of the embodiment of the present invention is a method for preparing a peptide composition described in JP-A-7-82299, a method for preparing a non-antigenic stabilizer described in JP-A-9-176196, or It can be prepared by the same method as the preparation method of the low antigenic stabilizer described in JP-A-11-12196. In general, the collagen component or the gelatin component is specifically decomposed using a collagenase enzyme. Is obtained.

  The wound healing agent according to the embodiment of the present invention is manufactured by a bioreactor method using a batch method or a column method using an immobilized collagenase enzyme in order to improve the enzyme recovery rate, using a gelatin component or a collagen component as a starting material. be able to. That is, collagenase enzyme is bound to various carriers such as chitopearl by physical adsorption method or chemical bonding method, and packed in a chromatography column, solubilized gelatin solution or decomposition does not occur at a constant temperature. Collagen denatured at a moderate temperature, preferably 40-45 ° C., is passed through the column for enzymatic degradation. In this case, the feeding rate of the collagen raw material is appropriately selected according to the activity of the immobilized enzyme and the required degree of degradation.

  A degradation product containing a peptide represented by the amino acid sequence (Gly-X—Y) n obtained by this adjustment method, preferably containing 5% or more of a collagen tripeptide, more preferably a collagen tripeptide, Wound healing agent, improvement against tendon damage or tear, healing promotion or prevention agent, tendon improvement or strengthening agent, improvement against ligament damage or tear, healing promotion or prevention agent, ligament improvement or strengthening agent, racehorse flexor tendon Inflammation, injury or tear improvement, healing promotion or prevention agent, racehorse flexor tendon improvement or strengthening agent, racehorse ligament injury or tear improvement, healing promotion or prevention agent, racehorse ligament improvement or enhancement It can be used as an agent.

  The effect of the wound healing agent according to the embodiment of the present invention is further increased by blending ascorbic acid or a derivative thereof into the peptide composition containing 5% or more of collagen tripeptide.

  The wound healing agent according to the embodiment of the present invention is digested and absorbed as quickly and efficiently as collagen or gelatin, which is known to be a protein that is easily digested and absorbed, and hydrolysates thereof. The

  The functional food according to an embodiment of the present invention includes 0.005% by mass or more of the wound healing agent. The functional food may be a health food, a health functional food such as a nutritional functional food or a food for specified health use, promote collagen production, promote osteoblast or fibroblast proliferation, strengthen bone In addition to functionalities such as density enhancement, vital tissue metabolism promotion, skin activation, prevention or improvement of functional deterioration due to aging of skin or bone, chronic rheumatism, osteoarthritis improvement and fracture healing promotion, tendon rupture, ligament Promotes healing of injuries, tendonitis, etc.

  The pharmaceutical of the embodiment of the present invention is characterized by containing 0.005% by mass or more of the wound healing agent. The category of pharmaceuticals includes quasi-drugs and cosmetics, promotes collagen production, promotes proliferation of osteoblasts or fibroblasts, strengthens or increases bone density, promotes metabolism of biological tissues, activates skin, skin or bone In addition to functionalities such as prevention or improvement of functional deterioration due to aging, chronic rheumatism, improvement of osteoarthritis and promotion of fracture healing, it has the ability to promote healing of tendon rupture, ligament injury, flexor tendonitis and the like.

  Collagen or gelatin, which is a starting material of the wound healing agent according to the embodiment of the present invention, includes bone, cartilage, skin, tendon, or shark, salmon, other fish, etc. such as cattle, pigs, birds, whales and other birds and beasts. What was prepared using fish skin, scales, etc. as a raw material can be used. Collagenase uses an enzyme that specifically cleaves the amino group side of glycine in the amino acid sequence unique to collagen: (Gly-X-Y) n.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this Example does not restrict | limit the scope of the present invention at all. Further, in the wound healing agent according to the present invention, a method of blending collagen tripeptide or collagen peptide containing 5% by mass or more of collagen tripeptide into functional foods and pharmaceuticals is performed in accordance with a method already known. be able to.

A wound healing agent was produced by the following method.
50 g of high-purity gelatin (manufactured by Zerais) was dissolved in 1,000 ml of 20 mM Tris-HCl buffer (pH 7.4) /0.1 M NaCl while heating, and then cooled to 50 ° C. The immobilized enzyme for enzymatic degradation was prepared by binding 100 mg of collagenase enzyme (purified from type IV, manufactured by Washington) to 50 g of particle carrier. The amount bound to the carrier was calculated by measuring the change in absorbance at 280 nm before and after the binding, and the binding rate was 99% or more. At the time of use, this immobilized enzyme was packed in a column type bioreactor and washed and equilibrated well with 20 mM Tris-HCl buffer (pH 7.4) /0.1 M NaCl buffer.

  Collagen or gelatin as a raw material was applied to the collagenase enzyme-immobilized column prepared in the above step, and enzymatic degradation was performed by the column method. During this time, the flow rate was controlled at 20-60 ml / min, and the column temperature was controlled at 39 ± 1 ° C. The enzyme reaction finished solution that came out of the column was collected and filtered through a 0.45 μm filter. The filtrate was pulverized by spray drying (spray dryer).

  The obtained collagen peptide having a collagen tripeptide content of 5% by mass or more (about 15% by mass in this example) was freeze-dried, and then the N-terminal amino acid was assayed by the Edman degradation method. As a result, 97.5% of the amino acids on the N-terminal side were found to be glycine, and it was proved to retain the (Gly-XY) n structure, which is an amino acid sequence characteristic of collagen according to the present invention. It was. The wound healing agent of this example is composed of the collagen peptide thus produced.

<Pharmacokinetic study>
Oral administration of the wound healing agent obtained in Example 1: collagen tripeptide labeled with radioisotope: tritium (3H) (hereinafter 3H-collagen tripeptide) or 3H-proline as a comparative control to rats. Then, the absorption and distribution of the test substance in the rat body were examined by (1) measurement of radioactivity concentration in plasma and (2) whole body autoradiography.

(1) Measurement of plasma radioactivity concentration As a result of measurement of plasma radioactivity concentration, plasma radioactivity concentration was already 1.4 μg eq./mL 5 minutes after oral administration of 3H-collagen tripeptide, Thereafter, it increased with time and reached the maximum concentration: 42.7 μg eq./mL. Subsequently, a gradual decrease was observed, and it was confirmed that the absorption rate was faster than that of the amino acid used as a control: 3H-proline.

(2) Whole body autoradiography After the rats administered with 3H-collagen tripeptide were killed by ether anesthesia, whole body autoradiogram was prepared, and the radioactivity distribution to various tissues and changes with time were examined. Whole body autoradiogram after 24 hours confirmed distribution to tendons, ligaments, bone marrow, periosteum, thymus, gastrointestinal tract, mucous membrane, skin and the like. In the whole body autoradiogram with 5 and 12 administrations, the distribution density was further observed, and the distribution to each tissue became clear. The whole body autoradiography is shown in FIG.

<Promoting collagen production>
To 5% FBS-Eagle MEM medium, 0 μM or 1 μM ascorbic acid (ASA), and the wound healing agent obtained in Example 1 at a concentration of 0 μg / mL or 3.0 μg / mL: collagen tripeptide added to the culture medium Then, normal human fibroblasts were cultured for 3 days in the culture solution. After completion of the culture, each culture supernatant was collected, and type I collagen secreted in each culture supernatant was measured by ELISA using rabbit anti-human type I collagen antiserum. As a result, as shown in FIG. 2, a. The ability to produce collagen is increased by adding the wound healing agent obtained in Example 1 alone, b. Collagen production is induced by adding only 1 μM ASA, c. It was revealed that the wound healing agent obtained in Example 1 further enhances collagen production induced by addition of ASA.

<Fracture healing promotion effect>
A fracture model rat was prepared by artificially fractured the left femur of a 7-week-old SD male rat. The wound healing agent obtained in Example 1 once a day for 12 weeks from the day after the model was prepared: Collagen tripeptide was orally administered with a sonde at 0,80,500 mg / kg. The day after the administration was completed, the femur (left and right) was removed, soft X-ray photography (using Softex-M-60, Softex-M-60) and bone strength measurement (using Malto-MZ500D) were performed. The healing state was determined.

(1) Soft X-ray photograph From the soft X-ray photograph (FIG. 3), the wound healing agent obtained by Example 1: collagen tripeptide administered group (80,500 mg / kg), the control group (0 mg / kg) ), The continuity of cortical bone increased, the fracture line tended to be thinner, and a tendency to promote fracture healing was confirmed.

(2) Measurement of bone strength When a three-point bending test (distance between fulcrums: 18 mm, front surface is upward, proximal side is right side and fractured at 20 mm / 60 s) was performed on the left and right femur shafts, a. Breaking time, b. Breaking displacement, c. Breaking load, d. Maximum load, e. For both parameters of stiffness, a large difference was observed in the control group (0 mg / kg) between the measurements of the left femur with and without the fracture treatment. In contrast, in the group (80,500 mg / kg) administered with the wound healing agent: collagen tripeptide obtained in Example 1, the difference was small, and it was restored to the same level as that of healthy bone (right femur). Was confirmed (see FIGS. 4 to 8).

<Healing promotion effect when Achilles tendon is torn>
The 7-week-old SD male rat left Achilles tendon was artificially cut to produce an Achilles tendon cut model rat, which was obtained according to Example 1 once a day for 2, 3 and 4 weeks. Wound healing agent: Collagen tripeptide was orally administered at 0.80 mg / kg with a sonde. On the next day after the administration end, the tendon around the Achilles tendon was taken out, and a tissue observation and a tensile test were performed to determine the degree of repair of the Achilles tendon.

(1) Tissue observation In the second week, the wound healing agent obtained in Example 1: collagen tripeptide administered group (80 mg / kg) showed more inflammation near the cut site than the control group (0 mg / kg). Suppression was observed, and vascular proliferation and a tendency to increase the amount of elastic fibers were observed. Furthermore, the number of osteoclasts in the tumor was small, and the shape of the bone and the state of the joint site between the tendon and the bone were close to the normal side (right), indicating a tendency for increased repair. At 4 weeks, most of the control group seemed to be restored, but the collagen tripeptide administration group was closer to normal.

(2) Tensile test A suture was wound around the tendon part and the muscle part, and the wound thread part was compressed and held, and further, a tensile test (50 mm / min) was carried out by pressing the tumor part. In the eyes, both the breaking load and the maximum load were high in the collagen tripeptide administration group (80 mg / kg), showing values closer to the normal side (right), and a tendency to promote repair was observed (FIG. 9).

Wound healing agent of Example 2 of the present invention: whole body auto showing the absorption and distribution in the rat body 24 hours after (A) 1 administration, (B) 5 administrations, (C) 12 administrations of collagen peptide It is a figure of radiography. In Example 3 of the present invention, human normal fibroblasts are cultured in (A) wound healing agent: a culture solution added with collagen peptide, and (B) wound healing agent: a culture solution added with collagen peptide and ascorbic acid, respectively. It is a graph which shows the density | concentration of the collagen secreted in the culture supernatant. For the fracture model rat artificially fractured in Example 4 of the present invention, wound healing agent: collagen tripeptide (group 1) 0 mg / kg (control group), (group 2) 80 mg / kg, ( (Group 3) Soft X-ray photographs of femurs (left and right) extracted after oral administration of 500 mg / kg. For the fracture model rat artificially fractured in Example 4 of the present invention, wound healing agent: collagen tripeptide (group 1) 0 mg / kg (control group), (group 2) 80 mg / kg, ( 3 group) is a graph showing the rupture time as a ratio of left and right femurs when a three-point bending test is performed on the diaphysis of femurs (left and right) extracted after oral administration of 500 mg / kg, respectively. For the fracture model rat artificially fractured in Example 4 of the present invention, wound healing agent: collagen tripeptide (group 1) 0 mg / kg (control group), (group 2) 80 mg / kg, ( 3 group) is a graph showing the fracture displacement as a ratio of left and right femurs when a three-point bending test is performed on the diaphysis of femurs (left and right) extracted after oral administration of 500 mg / kg. For the fracture model rat artificially fractured in Example 4 of the present invention, wound healing agent: collagen tripeptide (group 1) 0 mg / kg (control group), (group 2) 80 mg / kg, ( 3 group) is a graph showing the breaking load as a ratio of the left and right femurs when a three-point bending test is performed on the diaphysis of the femurs (left and right) extracted after oral administration of 500 mg / kg, respectively. For the fracture model rat artificially fractured in Example 4 of the present invention, wound healing agent: collagen tripeptide (group 1) 0 mg / kg (control group), (group 2) 80 mg / kg, ( 3 group) is a graph showing the maximum load as a ratio of left and right femurs when a three-point bending test is performed on the diaphysis of femurs (left and right) extracted after oral administration of 500 mg / kg. For the fracture model rat artificially fractured in Example 4 of the present invention, wound healing agent: collagen tripeptide (group 1) 0 mg / kg (control group), (group 2) 80 mg / kg, ( 3 group) is a graph showing the stiffness of a femoral bone (left and right) extracted after oral administration of 500 mg / kg, respectively, in a ratio of left and right femurs when a three-point bending test is performed on the diaphysis. The wound healing agent: Collagen tripeptide was orally administered at 0 mg / kg (control group) or 80 mg / kg to the Achilles tendon amputation model rat in which the Achilles tendon was artificially cut in Example 5 of the present invention. It is a graph which shows the breaking load when a tension test is implemented about a tendon, and a graph which shows the maximum load.

Claims (15)

A degradation product obtained by degrading collagen or gelatin with collagenase, wherein the amino acid sequence is (Gly-XY) n (where Gly represents a glycine residue, and X and Y are any amino acids other than glycine) Wound healing agent characterized by comprising as an active ingredient a peptide containing a peptide represented by the following formula: n represents a residue and n represents a positive integer. A degradation product obtained by degrading collagen or gelatin with collagenase, wherein the amino acid sequence is (Gly-XY) n (where Gly represents a glycine residue, and X and Y are any amino acids other than glycine) A wound healing agent comprising a peptide represented by the following formula: n represents a residue and n represents a positive integer. The wound healing agent according to claim 1 or 2, wherein the degradation product contains 5 mass% or more of a tripeptide whose amino acid sequence is represented by Gly-XY. A degradation product obtained by degrading collagen or gelatin with collagenase, wherein the amino acid sequence is Gly-XY (where Gly represents a glycine residue, and X and Y represent any amino acid residue other than glycine) A wound healing agent comprising a tripeptide represented by the formula: The wound healing agent according to any one of claims 1 to 4, wherein ascorbic acid or a derivative thereof is blended. An agent for improving, accelerating or preventing healing of tendon damage or tear, comprising the wound healing agent according to any one of claims 1 to 5. A tendon improving or reinforcing agent comprising the wound healing agent according to any one of claims 1 to 5. An agent for improving, healing, or preventing ligament damage or rupture, comprising the wound healing agent according to any one of claims 1 to 5. A ligament improving or reinforcing agent comprising the wound healing agent according to any one of claims 1 to 5. An agent for improving, promoting or preventing healing of inflammation, damage or rupture of a flexor tendon of a racehorse, comprising the wound healing agent according to any one of claims 1 to 5. An agent for improving or reinforcing a flexor tendon of a racehorse, comprising the wound healing agent according to any one of claims 1 to 5. An agent for improving, promoting or preventing healing of a ligament injury or rupture of a racehorse comprising the wound healing agent according to any one of claims 1 to 5. An agent for improving or strengthening a ligament of a racehorse, comprising the wound healing agent according to any one of claims 1 to 5. A functional food comprising 0.005% by mass or more of the wound healing agent according to any one of claims 1 to 5. A pharmaceutical comprising 0.005% by mass or more of the wound healing agent according to any one of claims 1 to 5.