KR102347731B1 - Composition for activating zinc transporter - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating osteoporosis comprising uridine. Specifically, the composition of the present invention increases Zip protein expression, BMP protein expression, and citrate production in osteoblasts to promote osteoblast differentiation, bone formation and bone length growth, thereby preventing, improving and treating osteoporosis and increasing height. It is characterized by being able to

Description

Zinc transporter activator composition {COMPOSITION FOR ACTIVATING ZINC TRANSPORTER}

The present invention relates to a composition for preventing, improving or treating osteoporosis, comprising uridine or a uridine derivative, and more particularly, by increasing Zip protein expression, BMP protein expression and citrate production in osteoblasts, osteoblast differentiation and It relates to a composition for preventing, improving or treating osteoporosis, characterized in that it promotes bone formation.

Bone is a dynamic tissue that is formed by osteoblasts and is destroyed and resorbed continuously and repeatedly by osteoclasts. Aged bone is removed and new tissue is formed through bone remodeling. Physiological bone remodeling occurs by the balanced action of bone formation and bone resorption. Diseases related to abnormal bone resorption in mammals including humans include bone degradation, malignant hypercalcemia, and metastatic bone disease, but the most common symptom of these diseases is osteoporosis.

Osteoporosis is a disease of the skeletal system in which bone strength is weakened and fractures easily occur. It is a disease caused by excessive bone resorption rather than bone formation as the balance between bone resorption and bone formation is disrupted. Bone strength is determined by the amount and quality of bone, and factors that affect bone quality include bone structure, replacement rate, mineralization, and micro-damage. Until now, there is no satisfactory indicator for overall evaluation of bone quality, so bone density is mainly used for diagnosing osteoporosis.

The causes of osteoporosis include aging, estrogen deficiency, hormonal changes, underweight, smoking, drugs, lack of exercise, and ovarian resection. Osteoporosis is particularly common in postmenopausal women. In addition, osteoporosis occurs due to diseases such as kidney, diabetes, hypertrophy of the thyroid gland, and early menopause. Osteoporosis is a problem with various fractures, especially femur fractures or vertebral fractures, which are easily caused by the weakening of bones rather than the symptoms themselves, and it makes long-term activities difficult, making it impossible to lead a healthy life. In addition, adolescent osteoporosis inhibits bone growth so that bone mass does not accumulate and the length of long bones (long bones) cannot grow, so height growth stops at an earlier age.

Until now, several substances have been developed to be applied to the treatment of osteoporosis, but none have been suggested as a fundamental treatment for osteoporosis, due to the insignificant therapeutic effect. Estrogen, which is most often used as a treatment for osteoporosis, has the disadvantage of having to continue taking it for the rest of your life. Absorption of alendronate from the digestive tract is slow, and there is a problem of causing inflammation in the gastrointestinal tract and esophageal mucosa. Although calcium preparations are known to have excellent effects with few side effects, it has been reported that they are not effective enough, and they are more of a preventive agent than a therapeutic agent. Because osteoporosis is a chronic disease, it is generally considered to require long-term treatment. Therefore, there is a need to develop a new therapeutic agent for osteoporosis, which has less toxicity and side effects and is effective in preventing and treating osteoporosis.

Uridine is a component of ribonucleic acid, and has been known for its preventive and therapeutic effects on arthritis, disc (distal disc), skin aging and wrinkles, dry eye syndrome, atopic dermatitis and dry skin (Korea Patent No. 10). -0977475) There is no known effect of uridine in the prevention and treatment of osteoporosis.

Zip protein (Zrt/Irt-like protein) is a zinc transporter protein that regulates zinc inflow and outflow into the cytoplasm. In humans, there are 14 Zip protein families. Zinc is stored in most bones in the human body and is not only a component of bone, but also an essential cofactor for proteins involved in microstructural stability and bone remodeling. do. In addition, citrate is an intermediate in the metabolic process (TCA cycle) that consumes energy using glucose, and is known as an important factor in the synthesis of bone matrix in osteoblasts.

BMP protein (Bone morphogenetic protein) is a protein with bone induction ability, and is a protein having various activities such as not only bone formation, but also early axis formation, nervous system formation, apoptosis, and cell differentiation. At least 15 types of BMP proteins have been discovered so far, and they are involved in bone growth by inducing stem cells to differentiate into the bone-forming lineage.

In this regard, the present inventors have found that uridine prevents and treats osteoporosis by increasing Zip protein expression, BMP protein expression, and citrate production in osteoblasts, thereby promoting osteoblast differentiation, bone formation, and growth of bone length, and Found that it is possible to grow the height, the present invention was completed.

Korean Patent Publication No. 10-0977475 (2010.08.23)

The present invention can prevent, treat or improve diseases such as osteoporosis and increase height by promoting osteoblast differentiation, bone formation and bone length growth by increasing Zip protein expression, BMP protein expression and citrate production in osteoblasts An object of the present invention is to provide a composition with

In order to achieve the above object, the present invention includes uridine, a uridine derivative or a pharmaceutically acceptable salt thereof, including osteoporosis, Paget's bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, It provides a pharmaceutical composition for preventing or treating at least one disease selected from the group consisting of dwarfism, dwarfism, and growth hormone deficiency.

In one embodiment, the uridine derivative is uracil, uridine monophosphate, uridine diphosphate, triacetyl uridine, tribenzoyl uridine, 5-ethyl uridine, 2-deoxyuridine and isopropylidene uridine. It may be one or more selected from the group consisting of.

In one embodiment, the composition may include one or more of zinc or aspartic acid. In addition, the zinc and aspartic acid may be included in the form of a complex.

In one embodiment, the composition may further comprise vitamin D.

In one embodiment, the composition may promote expression of a Zip protein (Zrt/Irt-like protein) in osteoblasts, and the Zip protein may be a Zip1 protein or a Zip13 protein.

In one embodiment, the composition may promote the expression of a bone morphogenetic protein (BMP), and the BMP protein may be a BMP2 protein.

In one embodiment, the composition may increase citrate production.

In addition, the present invention, including zinc and aspartic acid, osteoporosis, Paget's bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, dwarfism, dwarfism, and selected from the group consisting of growth hormone deficiency Provided is a pharmaceutical composition for preventing or treating one or more diseases.

In one embodiment, the composition may further comprise vitamin D.

In one embodiment, the composition may promote expression of a Zip protein (Zrt/Irt-like protein) in osteoblasts, and the Zip protein may be a Zip1 protein or a Zip13 protein.

In one embodiment, the composition may promote the expression of a bone morphogenetic protein (BMP), and the BMP protein may be a BMP2 protein.

In one embodiment, the composition may increase citrate production.

In addition, the present invention, including uridine, uridine derivatives or salts thereof, osteoporosis, Paget's bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, dwarfism, dwarfism, and growth hormone deficiency It provides a food composition for preventing or improving one or more diseases selected from the group consisting of.

In one embodiment, the uridine derivative is uracil, uridine monophosphate, uridine diphosphate, triacetyl uridine, tribenzoyl uridine, 5-ethyl uridine, 2-deoxyuridine and isopropylidene uridine. It may be one or more selected from the group consisting of.

In one embodiment, the composition may include one or more of zinc or aspartic acid. In addition, the zinc and aspartic acid may be included in the form of a complex.

In one embodiment, the composition may further comprise vitamin D.

In addition, the present invention, including uridine, uridine derivatives or salts thereof, osteoporosis, Paget's bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, dwarfism, dwarfism, and growth hormone deficiency It provides a feed composition for animals for preventing or improving one or more diseases selected from the group consisting of.

In one embodiment, the uridine derivative is uracil, uridine monophosphate, uridine diphosphate, triacetyl uridine, tribenzoyl uridine, 5-ethyl uridine, 2-deoxyuridine and isopropylidene uridine. It may be one or more selected from the group consisting of.

In one embodiment, the composition may include one or more of zinc or aspartic acid. In addition, the zinc and aspartic acid may be included in the form of a complex.

In one embodiment, the composition may further comprise vitamin D.

In addition, the present invention, including zinc and aspartic acid, osteoporosis, Paget's bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, dwarfism, dwarfism, and selected from the group consisting of growth hormone deficiency It provides a food composition for preventing or ameliorating one or more diseases.

In addition, the present invention, including zinc and aspartic acid, osteoporosis, Paget's bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, dwarfism, dwarfism, and selected from the group consisting of growth hormone deficiency It provides a feed composition for animals for preventing or improving one or more diseases.

The composition according to the present invention promotes osteoblast differentiation, bone formation and growth of bone length by increasing Zip protein expression, BMP protein expression, and citrate production in osteoblasts, and has the effect of preventing bone loss, It can be usefully used to prevent, improve or treat diseases such as jet bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, dwarfism, dwarfism and growth hormone.

1 shows the cancellous bone volume (BV/TV) and cancellous bone thickness (Tb. ) and bone density (BMD).

Hereinafter, embodiments and examples of the present invention will be described in detail with reference to the accompanying drawings so that those of ordinary skill in the art to which the present invention pertains can easily carry out. However, the present application may be embodied in various forms and is not limited to the embodiments and examples described herein.

Throughout this specification, when a part "includes" a component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.

The present invention relates to osteoporosis, Paget's bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, It relates to a composition for preventing, improving or treating at least one disease selected from the group consisting of dwarfism, dwarfism, and growth hormone deficiency.

[Formula 1]

The composition may be a pharmaceutical composition, a food composition, or a feed composition for animals.

In one embodiment, the uridine derivative is uracil, uridine monophosphate, uridine diphosphate, triacetyl uridine, tribenzoyl uridine, 5-ethyl uridine, 2-deoxyuridine and isopropylidene uridine. It may be one or more selected from the group consisting of.

In one embodiment, the composition may further include at least one of zinc and aspartic acid, and the zinc and aspartic acid may be included in the form of a complex. In one embodiment, the complex may be a simple mixture of zinc and aspartic acid, and in another embodiment, the complex may be in the form of a chelate of zinc and aspartic acid. A method for preparing the chelate compound of zinc and aspartic acid is not particularly limited and may be prepared by a method known in the art. For example, it can be obtained by adding aspartic acid to the seaweed calcium dispersion, stirring it until it dissolves, adding zinc sulfate and reacting sufficiently, and then freeze-drying the supernatant obtained by centrifugation.

In one embodiment, the composition may further comprise vitamin D. Vitamin D is a fat-soluble vitamin that is a component of bones and teeth. Vitamin D is an essential element of calcium metabolism that helps the small intestine absorb calcium, accumulates calcium absorbed in the body in bones and teeth, promotes calcium reabsorption in the kidneys, and maintains adequate blood calcium concentration. It helps in the accumulation of height and is also involved in height growth. Vitamin D is incorporated in most calcium supplements, and deficiency can lead to rickets and osteoporosis.

In one embodiment, the composition may promote Zrt/Irt-like protein (Zrt/Irt-like protein) expression and bone morphogenetic protein (BMP) expression or increase citrate production in osteoblasts.

Osteoblasts (osteoblasts) are cells that appear during bone development or regeneration and are involved in the formation of the bone matrix. Bone matrix is composed of collagen fibers and an amorphous matrix occupying the interfiber. Osteoblasts line up around the bone being formed to create a bone matrix around the bone. Secondary calcium phosphate deposits in the bone matrix to form bone. Some of the osteoblasts are buried in the bone matrix created by themselves and become osteocytes. When bone growth is over, osteoblasts stop their activity and show a form indistinguishable from fibroblasts.

Zip protein (Zrt/Irt-like protein) is a zinc transporter protein that regulates zinc inflow and outflow into the cytoplasm of osteoblasts. It is an essential cofactor for the proteins involved.

Bone morphogenetic protein (BMP) is a protein that has the ability to induce bone and grow bone length, and is involved in bone growth by inducing stem cells to differentiate into osteogenic lineages.

Citrate is an intermediate in the metabolic process (TCA cycle) that uses glucose to consume energy. Citrate accounts for 1-2% of bone weight, and the concentration of citrate in bone is 5 to 25 times higher than that of other tissues, which confer important properties such as stability, strength and resistance to bone. In addition, citrate is an important factor in the synthesis of bone matrix in osteoblasts.

Therefore, promoting the expression of Zip protein and BMP protein and increasing the amount of citrate production promotes osteoblast differentiation, bone formation, and growth of bone length, which means that it has an effect in preventing or treating osteoporosis and in height growth.

As used herein, "pharmaceutically acceptable salt" refers to a salt that is pharmaceutically acceptable and retains the desired pharmacological activity of the parent compound. The salt is not particularly limited as long as it is pharmaceutically acceptable, and for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, maleic acid, succinic acid, methanesulfonic acid , benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, and the like can be used.

The pharmaceutical composition of the present invention may be administered parenterally or orally according to a desired method, and the dosage may vary depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of disease, etc. Its scope is diverse. In addition, the therapeutically effective amount of the composition may vary depending on the method of administration, the target site, and the condition of the patient.

In one embodiment, the pharmaceutical composition of the present invention is a powder, granules, tablets, soft or hard capsules, suspensions, emulsions, syrups, aerosols, etc. oral dosage forms, ointments, creams, etc., according to a conventional method, respectively. , suppositories, injections and sterile injection solutions, etc., may be formulated and used in any form suitable for pharmaceutical preparations.

Excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and diluents commonly used for the formulation may be further included. For example, excipients that may be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate , cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate, mineral oil and the like may be used, but is not limited thereto. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a suppository base, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, etc. can be used.

The food composition of the present invention may be a health functional food, and the "health functional food" means a food manufactured and processed using raw materials or ingredients useful for the human body, and "functionality" means the structure and function of the human body It refers to ingestion for the purpose of obtaining useful effects for health purposes, such as regulating nutrients or physiological effects.

Food compositions can be manufactured and processed into health functional foods in the form of tea bags, leached teas, health drinks, etc., and can also be processed into other breads, confectionery, ice cream, noodles, and the like. In addition, the above 　 food 　 composition may be a health supplement 　 or a 　 food additive, and the suitability as a "food additive" is evaluated on the item according to the general rules and general test methods of the Food Additives Code, etc. approved by the Ministry of Food and Drug Safety, unless otherwise specified. It is judged according to the relevant standards and standards.

The present invention will be described in more detail through the following examples, but the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[Example 1] Confirmation of cancellous bone volume, thickness and bone density improvement effect

The prevention and improvement effects of osteoporosis according to uridine treatment were confirmed using 6-week-old female rats (Sprague-Dawley) purchased from Orient Bio (Seongnam, Korea).

When the average weight of the rats reached about 150 g, the experiment was performed by dividing the mice into three experimental groups under the following conditions.

(i) Most surgical group without ovarian resection (Sharm)

(ii) Ovariectomy group (OVX-Ctrl)

(iii) Test group (OVX-Uridine): uridine 75mg/kg administered for 20 weeks after ovariectomy

The breeding room environment was maintained at a temperature of 22 ± 2 °C and a humidity of 65 ± 5%, and the light and dark was controlled in a 12-hour cycle (light: 06:00 to 18:00). Thereafter, Trabecular bone volume (BV/TV), Trabecular Thikness (Tb.Th), and Bone Mineral Density (BMD) were measured from the shin bone (Tibia) and shown in FIG. 1 .

As shown in FIG. 1 , the test group administered with uridine after ovariectomy showed superior effects in all of cancellous bone volume, thickness, and bone density compared to the ovariectomy group not administered with uridine. It was confirmed that the increase was even greater compared to the most operated group without resection.

Therefore, it can be confirmed that uridine has an excellent effect of inhibiting bone loss as well as an effect of increasing bone formation and bone density.

[Example 2] Confirmation of promotion of Zip1, Zip13 and BMP2 mRNA expression in osteoblasts

An experiment to confirm the effect of uridine and zinc-aspartic acid complex treatment on the expression of Zip1 mRNA, Zip13 mRNA and BMP2 mRNA in osteoblasts was performed as follows.

In 10% FBS in DMEM medium, osteoblastic progenitor cells MC3T3-E1 Subclone 4 (ATCC ^® CRL-2593TM) were aliquoted into 6 wells at 2 x 10 ⁴ cells/well. 2 days after cell confluence, differentiation medium (10% FBS, 1% penicillin, 1:1000 L-ascorbic acid (Sigma Aldrich), 1:100 β-glycerophosphate disodium salt hydrate (Sigma Aldrich) in DMEM) , uridine, zinc-aspartic acid complex, and a 1:1 mixture of uridine and zinc-aspartic acid complex were treated, respectively, and the medium was exchanged for 6 days to induce osteoblast differentiation. Thereafter, cells were harvested and RNA was isolated using the NucleoSpin RNA Kit (MACHEREY-NAGEL, 740955.50). After quantifying the isolated RNA, it was converted into cDNA using ReverTra Ace q PCR RT Master Mix (TOYOBO, FSQ-201).

The sense strand primer and anti-sense strand primer of the target genes Zip1 mRNA, Zip13 mRNA, and BMP mRNA were put together, and real-time PCR (ViiA7, AB life techonolgies) was performed for each mRNA. After amplification for each cycle, the number of cycle repetitions was determined by observing the cycle corresponding to the effective concentration curve. Using the expression level of the GAPDH gene, osteoblasts (ND) and osteoblasts (OD) not treated with the test substance, The relative gene expression level of the target gene for the test substance treatment group was calculated and shown in Table 1 below.

mRNA
(relative expression) ND
(osteoblasts) OD
(osteoblasts) zinc-asp complex
5 μg/ml zinc-asp complex
10 μg/ml Uridine
5 μg/ml Uridine+
zinc-asp (1:1) complex
5 μg/ml Zip1
1.00 ± 0.21 ^*** 1.22 ± 0.20 1.30 ± 0.19 1.55 ± 0.15 ^*** 1.32 ± 0.17 1.62 ± 0.12 ^*** Zip13 1.00 ± 0.02 ^*** 3.00 ± 0.25 3.38 ± 0.015 3.99 ± 0.28 ^*** 3.27 ± 0.11 3.86 ± 0.09 ^*** BMP2 1.00 ± 0.21 ^*** 1.91 ± 0.20 1.95 ± 0.05 3.25 ± 0.11 ^*** 2.90 ± 0.14 ^** 3.55 ± 0.13 ^***

*<0.05, **<0.01, ***<0.001 vs OD (Osteogenic differentiation)

As shown in Table 1, the expression of Zip1, Zip13 and BMP2 mRNA was significantly increased in the group treated with uridine or zinc-aspartic acid (zinc-asp) complex compared to OD, and uridine and zinc-aspartic acid complex In the group treated with , each mRNA expression level was further increased compared to the group treated with the same amount.

Therefore, uridine can promote the expression of Zip1 protein, Zip13 protein and BMP2 protein in osteoblasts to promote osteoblast differentiation, bone formation, and bone length growth. If you do, you can see that it rises even more.

[Example 3] Confirmation of promotion of citrate production in osteoblasts

To determine whether citrate production is promoted in osteoblasts when uridine and zinc-aspartic acid complex are treated, citrate production was measured using a citrate assay kit (ab83396, Cambridge, Cambridgeshire, UK).

In a differentiation medium, osteoblast MC3T3-E1 cells were treated with uridine and/or zinc-aspartic acid complex and cultured for 6 days, followed by cell lysis. At this time, the assay buffer contained in the kit was added and pipetted to mix the cells and the buffer well, and centrifuged at 16,000 g x 5 min x 4 °C, and the supernatant was collected. The measured values were corrected to values quantified using the Bradford protein assay method, and are shown in Table 2 below.

ND
(osteoblasts) OD
(osteoblasts) zinc-asp complex
5 μg/ml zinc-asp complex
10 μg/ml Uridine
5 μg/ml Uridine+
zinc-asp (1:1) complex
5 μg/ml citrate
(nmol/mg protein) 0.52 ± 0.02 ^** 1.32 ± 0.09 1.45 ± 0.12 ^* 1.67 ± 0.18 ^** 1.42 ± 0.11 ^* 1.81 ± 0.13 ^***

*<0.05, **<0.01, ***<0.001 vs OD (Osteogenic differentiation)

As shown in Table 2, the uridine or zinc-aspartic acid (zinc-asp) complex treatment group significantly increased the amount of citrate compared to the OD, and the uridine and zinc-aspartic acid complexes were treated together. The group had the highest production.

Therefore, uridine can increase osteoblast differentiation and bone formation by promoting the production of citrate, and it can be confirmed that these effects are further enhanced when uridine and zinc-aspartic acid complex are treated together.

Claims (19)

a) uridine, a uridine derivative or a pharmaceutically acceptable salt thereof; and
b) zinc or aspartic acid;
The uridine derivative is selected from the group consisting of uracil, uridine monophosphate, uridine diphosphate, triacetyl uridine, tribenzoyl uridine, 5-ethyl uridine, 2-deoxyuridine and isopropylidene uridine. characterized in that one or more,
Osteoporosis, Paget's bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, dwarfism, dwarfism, and a pharmaceutical composition for the prevention or treatment of one or more diseases selected from the group consisting of growth hormone deficiency.
delete delete The pharmaceutical composition according to claim 1, further comprising vitamin D.
The pharmaceutical composition according to claim 1, wherein the composition promotes the expression of Zip protein (Zrt/Irt-like protein) in osteoblasts.
The pharmaceutical composition according to claim 5, wherein the Zip protein is a Zip1 protein or a Zip13 protein.
The pharmaceutical composition of claim 1 , wherein the composition increases the production of citrate.
The pharmaceutical composition according to claim 1, wherein the composition promotes BMP2 protein (Bone morphogenetic protein 2) expression.
delete a) uridine, uridine derivatives or salts thereof; and
b) zinc or aspartic acid;
The uridine derivative is selected from the group consisting of uracil, uridine monophosphate, uridine diphosphate, triacetyl uridine, tribenzoyl uridine, 5-ethyl uridine, 2-deoxyuridine and isopropylidene uridine. characterized in that one or more,
Osteoporosis, Paget's bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, dwarfism, dwarfism, and a food composition for the prevention or improvement of one or more diseases selected from the group consisting of growth hormone deficiency.
delete delete 11. The food composition of claim 10, further comprising vitamin D.
a) uridine, uridine derivatives or salts thereof; and
b) zinc or aspartic acid;
The uridine derivative is selected from the group consisting of uracil, uridine monophosphate, uridine diphosphate, triacetyl uridine, tribenzoyl uridine, 5-ethyl uridine, 2-deoxyuridine and isopropylidene uridine. characterized in that one or more,
Osteoporosis, Paget's bone disease, rickets, osteomalacia, alveolar bone loss, osteopenia, renal osteodystrophy, short stature, dwarfism, dwarfism, and at least one disease selected from the group consisting of growth hormone deficiency, animal feed composition for the prevention or improvement.
delete delete 15. The animal feed composition according to claim 14, further comprising vitamin D. delete delete

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