COMPOSITION FOR GROWTH PROMOTION, CONTAINING COUMARIC ACID AS ACTIVE INGREDIENT

Abstract

The present invention relates to a growth promotion effect of coumaric acid and, more specifically, to a pharmaceutical composition, a food composition, and an animal feed composition, which are for growth promotion and contain coumaric acid as an active ingredient. The composition containing coumaric acid as an active ingredient, of the present invention, promotes chondrocyte proliferation and growth hormone secretion, thereby finally having an effect of promoting growth, and thus the composition of the present invention is effective in not only growth and skeletal formation of growing children and adolescents, but also height growth therapy by being used alone or in combination with growth hormone therapy.

Claims

1. A method for pormoting growth, the method comprising administering to a subject in need thereof an effective amount of coumaric acid as an active ingredient.

2-3. (canceled)

4. The method according to claim 1, wherein the coumaric acid is any one or more selected from the group consisting of o-coumaric acid, m-coumaric acid, and p-coumaric acid.

5. The method according to claim 1, wherein the growth is bone growth.

6-9. (canceled)

Description

BRIEF DESCRIPTION OF DRAWINGS

[0048] FIG. 1 is a diagram illustrating changes in body weight of rats in administration groups for 10 days.

[0049] FIG. 2 is a graph illustrating total tibial length (Data represent mean (n=5), *p<0.05).

[0050] FIG. 3A is a graph illustrating the height of the growth plate of rats of each administration group, and FIGS. 3B and 4C are graphs illustrating the heights of a proliferative zone and a hypertrophic zone (A: 20 magnification, *p<0.05 and ***p<0.001).

[0051] FIG. 4A is an image of the proximal tibia growth plate, stained with BrdU, in rats of each administration group, and FIG. 5B is a graph illustrating the ratio of BrdU-labeled cartilage cells in rats of each administration group (A: 40 magnification, *p<0.05).

[0052] FIG. 5 is a graph illustrating the growth hormone levels in the blood of rats in each administration group (Data represent mean (n=5), *p<0.05).

[0053] FIG. 6A is an image of the proximal tibia growth plate, stained with BrdU, in rats of each administration group, and FIG. 6B is a graph illustrating the levels of IGF-1 in the blood of rats in each administration group (A: 20 magnification, B: **p<0.01 and ***p<0.001).

DETAILED DESCRIPTION OF EMBODIMENT

[0054] Hereinafter, this invention will be described in detail.

[0055] However, the following examples are merely provided to exemplify this invention, not to limit the scope of this invention.

EXPERIMENTAL EXAMPLE 1

[0056] Assessment of Growth Promoting Effect of Coumaric Acid Using Animal Models

[0057] <1-1> Administration of Coumaric Acid

[0058] Three-week-old male Sprague-Dawley rats were purchased and acclimated for three days. A breeding environment was altered to a 24-hour pattern, a light/dark cycle was maintained at 12-hour intervals, and antibiotic-free normal solid feed was used. In a control group, saline was injected. In a positive control group, Eutropin Injection (LG Bioscience Korea) was subcutaneously injected at a dose of 20 g/kg once a day. The coumaric acid of this invention was administered to the oral cavity of all rats of a coumaric acid group, once daily. The drug administered to each group is as follows:

[0059] Control: normal saline, 1 ml

[0060] Positive Control: growth hormone, 20 g/kg

[0061] Coumaric acid: 100 mg/kg

[0062] After feeding and administering the coumaric acid to the rats for 10 days, autopsies were carried out to confirm the following content.

[0063] <1-2> Changes in the Body Weight of Laboratory Animals

[0064] Body weights before feeding and body weights after feeding for 10 days were measured for each group of Experimental Example <1-1>. The measurement was carried out twice a week. The measured values were statistically analyzed to calculate meanstandard deviation.

[0065] As shown in FIG. 1, the 10-day observation showed that there were almost no difference between the body weights in all of the control group, the experimental group, and growth hormone-administered group. It shows that there is no close correlation between the body weight of a mouse and the effect of coumaric acid administration.

[0066] <1-3> Measurement of Changes in the Total Tibial Length of Laboratory Animals

[0067] To assess the effect of coumaric acid on growth promotion, total tibial length of rats were measured. In Experimental Example <1-1>, after the 10-day administration was completed, right and left tibias (shinbones) were extracted from the control groups and the coumaric acid-fed experimental group according to this invention, all the muscles, fat and ligaments attached to bone tissue were removed, and then the resulting specimen was stored in 70% alcohol, followed by radiographic analysis using an X-ray imaging instrument (OM-FORTE-10121, DK Medical Systems). For the analysis, 55 kV, 320 A X-ray sources were used. The total tibial length was measured by X-ray radiography.

[0068] As shown in FIG. 2, it can be seen that the total tibia length of the growth hormone-administered group was greater than the control group, and the total tibial length of the coumaric acid-administered group was also greater than the control group.

[0069] <1-4> Analysis of Growth Plate in Laboratory Animals

[0070] To assess the growth effect of coumaric acid, the growth plate of each rat was analyzed. In Experimental Example <1-1>, after the 10-day administration was completed, right and left tibias (shinbones) were extracted from the experimental group and the control groups according to this invention, all the muscles, fat and ligaments attached to bone tissue were removed, and then the resulting specimen was stored in 70% alcohol, demineralized, and then dehydrated. Afterward, the resulting specimen was washed with xylene, and then put into a paraffin block. Subsequently, the specimen was cut to a thickness of 4 m, and then the heights of the proliferative zone and the hypertrophic zone in the growth plate were measured using a BX50 microscope (Olympus).

[0071] As shown in FIG. 3, on average, the height of the proliferative zone of the control group was 0.77 mm, and the height of the proliferative zone of the growth hormone-administered group was 1.13 mm. In addition, the height of the proliferative zone of the coumaric acid-administered group was measured at 0.98 mm. It was confirmed that both of the growth hormone-administered group and the coumaric acid-administered group showed significant increases in the height of the proliferative zone, compared to the control group, and particularly, the increase in the height of the proliferative zone of the coumaric acid-administered group was similar to that of the growth hormone-administered group. It was also confirmed that the height of the hypertrophic zone was 1.44 mm in the growth hormone-administered group, and 1.27 mm in the coumaric acid-administered group, both of which were greater than that of the control group (1.10 mm).

[0072] <1-5> Observation of Proliferation of Cartilage Cells in Proximal Tibia in Laboratory Animals

[0073] To confirm whether the effect of coumaric acid on bone growth is caused by the proliferation of cartilage cells, cartilage cells were observed. In Experimental Example <1-1>, after the 10-day administration was completed, rats of the experimental group and the control groups according to this invention were intraperitoneally administered with a dose of 30 mg/kg of 5-bromo-2-deoxyuridine (BrdU) on the day of the experiment. Two hours later, autopsies were carried out to extract legs, and then the legs were stored in 4% PFA for 24 hours. Following embedding and sectioning, cartilage cells which were stained using a BrdU staining kit (Invitrogen, Carlsbad, Calif., USA) were observed.

[0074] As shown in FIG. 4, a ratio of the BrdU-labeled cells with respect to total cells was 0.13% in the control group, 0.18% in the growth hormone-administered group, and 0.18% in the coumaric acid-administered group. In other words, the ratios of the BrdU-labeled cartilage cells were increased in both of the growth hormone-administered group and the coumaric acid-administered group, compared to the control group.

[0075] <1-6> Measurement of Growth Hormone Levels in Blood of Laboratory Animals

[0076] To confirm whether coumaric acid affects the secretion level of the growth hormone, a growth hormone level in the blood of each rat was measured. In Experimental Example <1-1>, after the 10-day administration was completed, blood was collected from the inferior vena cava of the rats in the experimental group and the control groups according to this invention on the day of the experiment, and then centrifuged to obtain serum. The measurement of the growth hormone levels were carried out using a Growth Hormone Rat ELISA Kit (Life Technologies, KRC5311).

[0077] As shown in FIG. 5, the growth hormone levels in blood were 3.97 ng/ml in the control group, 11.89 ng/ml in the growth hormone-administered group, and 7.62 ng/ml in the coumaric acid-administered group, which showed that the growth hormone levels were significantly increased in both of the growth hormone-administered group and the coumaric acid-administered group, compared to the control group.

[0078] <1-7> Measurement of IGF-1 Protein Levels in Blood of Laboratory Animals

[0079] To confirm whether coumaric acid affects the secretion levels of IGF-1, which is a protein that stimulates growth hormone secretion, IGF-1 protein levels were measured. In Experimental Example <1-1>, after the 10-day administration was completed, the rats of the experimental group and the control groups according to this invention were anesthetized on the day of the experiment, and then blood was extracted therefrom using a syringe and centrifuged to obtain serum. The measurement of the IGF-1 levels was carried out using an IGF1 ELISA Kit (Abcam, ab100695).

[0080] As shown in FIG. 6, the IGF-1 protein levels were 0.84 mg/kg in the control group, 1.31 mg/kg in the growth hormone-administered group, and 1.23 mg/kg in the coumaric acid-administered group, which showed that the IGF-1 protein levels were significantly increased in both of the growth hormone-administered group and the coumaric acid-administered group, compared to the control group.

INDUSTRIAL AVAILABILITY

[0081] As described above, this invention relates to a growth promoting effect of coumaric acid, and more particularly, to a pharmaceutical composition, a food composition and an animal feed composition, which contain coumaric acid as an active ingredient, for promoting growth.

[0082] The compositions containing coumaric acid an active ingredient according to this invention have an effect of promoting the proliferation of cartilage cells and growth hormone secretion, resulting in the promotion of growth. Therefore, the compositions of this invention are effective for the growth and skeletal formation of children and adolescents in a growing phase, and also effective in height growth treatment when used alone or in combination with growth hormone therapy, and thus have high industrial availability.