WO2015050160A1

WO2015050160A1 - Anti-aging pharmaceutical composition for muscle containing as active ingredient parathyroid hormone or derivative thereof

Info

Publication number: WO2015050160A1
Application number: PCT/JP2014/076289
Authority: WO
Inventors: 圭祐萩原
Original assignee: 国立大学法人大阪大学
Priority date: 2013-10-03
Filing date: 2014-10-01
Publication date: 2015-04-09

Abstract

The present invention provides an anti-aging pharmaceutical composition for muscle containing as the active ingredient parathyroid hormone, a parathyroid hormone derivative, or a salt thereof. Preferably the parathyroid hormone derivative is teriparatide. The anti-aging pharmaceutical composition is capable of preventing age-related reduction in muscular strength and preventing or treating sarcopenia and locomotive syndrome.

Description

Pharmaceutical composition for preventing aging of muscle, comprising parathyroid hormone or a derivative thereof as an active ingredient

The present invention relates to a pharmaceutical composition for preventing muscle aging, which comprises parathyroid hormone or a derivative thereof as an active ingredient.

In recent years, the decrease in muscle strength and muscle mass associated with aging is called “Sarcopenia” and has attracted much attention as being related to bedridden. It is said that sarcopenia involves various factors such as nutrient intake, hormones, and inflammatory response in addition to the decrease in physical activity that changes with aging, and it is estimated that 50% or more are affected at age 80 and over. When sarcopenia occurs, it causes a vicious circle of easy falls / falls, fractures, body movement limitations, and progression of sarcopenia, and induces a bedridden state. There is a report that mortality rate is 2.3 times higher when sarcopenia is merged at the age of 70 or more than when not merging. According to the welfare white paper, it is estimated that the number of bedridden elderly people will reach 2.3 million in 2025 in Japan, and prevention and treatment of sarcopenia is an important issue.

At present, the most effective means for prevention and treatment of sarcopenia is strength training (Non-Patent Documents 1 and 2), but there are complications such as joints and circulatory organs, and body movement is already restricted. If so, it is difficult to exercise with sufficient intensity. Clinical studies on essential amino acids, hormonal agents (testosterone, estrogen, growth hormone) and ACE inhibitors have been conducted, but amino acids have no therapeutic effect (Non-patent Document 3), and hormonal agents such as breast cancer and prostate cancer It has been reported that the risk increases and ACE inhibitors have few cases and do not lead to evidence construction (Non-patent Document 4). In other words, at present, sufficient measures have not been taken against sarcopenia, and there is no drug approved as a therapeutic drug.

Also, the concept of “locomotive syndrome” has been proposed by the Japanese Orthopedic Association. Locomotive syndrome refers to a condition in which the mobility function has deteriorated due to a disorder of the musculoskelet. More specifically, one or more of the musculoskeletal organs such as muscle, bone, joint, cartilage, and intervertebral disc have failed. This refers to a state where some obstacles are caused to walking and daily life. As the locomotive syndrome progresses, the risk of needing care increases, so it is considered important to prevent locomotive syndrome and extend healthy life expectancy. Sarcopenia is considered one of the most important factors causing locomotive syndrome, therefore preventing sarcopenia and improving muscle weakness and loss of muscle slows the progression of locomotive syndrome and prevents locomotive syndrome And greatly contribute to improvement.

On the other hand, natural human parathyroid hormone (PTH) is a polypeptide consisting of 84 amino acids secreted from the parathyroid gland, and its main physiological functions are bone resorption and bone formation in the bone, and phosphorus resorption in the kidney. Suppression of calcium and promotion of calcium reabsorption. The biological activity of PTH is on the N-terminal side of the structure, and the 1-34 peptide chain has been shown to have almost all of the biological activities of PTH to the same extent.
The N-terminal fragment 1-34 of PTH is referred to as teriparatide and is widely used as an osteoporosis therapeutic agent (Asahi Kasei Pharma “Teribbon®”, Eli Lilly “Forteo®”). In addition, it has been reported that teriparatide has a strong fracture-suppressing action, which is considered to be caused by an increase in bone density caused by teriparatide administration (Non-patent Document 5). However, the effect of teriparatide on muscle has not been reported.

An object of the present invention is to provide a pharmaceutical composition useful for preventing muscle aging, particularly a pharmaceutical composition useful for preventing or treating sarcopenia and locomotive syndrome by suppressing the decrease in muscle strength accompanying aging.

The present invention includes the following inventions in order to solve the above problems.
[1] A pharmaceutical composition for preventing muscle aging, which contains parathyroid hormone, a parathyroid hormone derivative or a salt thereof as an active ingredient.
[2] The pharmaceutical composition according to [1], wherein the parathyroid hormone derivative is teriparatide.
[3] The pharmaceutical composition according to the above [1] or [2], which is for suppressing muscle weakness.
[4] The pharmaceutical composition according to any one of the above [1] to [3], which is used for prevention and / or treatment of sarcopenia.
[5] The pharmaceutical composition according to any one of [1] to [3], which is used for prevention and / or treatment of locomotive syndrome.

[6] A method for preventing muscle aging, comprising administering an effective amount of a parathyroid hormone, a parathyroid hormone derivative or a salt thereof to a mammal.
[7] A method for suppressing muscle weakness, comprising administering an effective amount of a parathyroid hormone, a parathyroid hormone derivative or a salt thereof to a mammal.
[8] A method for preventing and / or treating sarcopenia, comprising administering an effective amount of parathyroid hormone, a parathyroid hormone derivative or a salt thereof to a mammal.
[9] A method for preventing and / or treating locomotive syndrome, comprising administering an effective amount of parathyroid hormone, a parathyroid hormone derivative or a salt thereof to a mammal.
[10] Use of parathyroid hormone, a parathyroid hormone derivative or a salt thereof for producing a pharmaceutical composition for preventing muscle aging.
[11] Use of parathyroid hormone, a parathyroid hormone derivative or a salt thereof for producing a pharmaceutical composition for suppressing muscle weakness.
[12] Use of a parathyroid hormone, a parathyroid hormone derivative or a salt thereof for producing a pharmaceutical composition for preventing and / or treating sarcopenia.
[13] Use of a parathyroid hormone, a parathyroid hormone derivative or a salt thereof for producing a pharmaceutical composition for preventing and / or treating locomotive syndrome.
[14] A parathyroid hormone, a parathyroid hormone derivative or a salt thereof for use in preventing muscle aging.
[15] A parathyroid hormone, a parathyroid hormone derivative, or a salt thereof for use in suppressing a decrease in muscle strength.
[16] A parathyroid hormone, a parathyroid hormone derivative or a salt thereof for use in the prevention and / or treatment of sarcopenia.
[17] A parathyroid hormone, a parathyroid hormone derivative or a salt thereof for use in the prevention and / or treatment of locomotive syndrome.

According to the present invention, a pharmaceutical composition useful for preventing muscle aging can be provided. The pharmaceutical composition of the present invention can suppress a decrease in muscle strength associated with aging, and is very useful as a pharmaceutical for preventing or treating sarcopenia and a pharmaceutical for preventing or treating locomotive syndrome.

It is a figure which shows the average body weight of each group at the age of 20 weeks in the teriparatide administration test using an aging promotion model mouse (SAMP8). In a teriparatide administration test using an aging promoting model mouse (SAMP8), it is a figure showing the average body weight of each group at the age of 30 weeks. In the teriparatide administration test using an aging promoting model mouse (SAMP8), it is a figure showing the results of bone mass measurement by 2D-CT of representative individuals in each group at the age of 20 weeks. In a teriparatide administration test using an aging promoting model mouse (SAMP8), it is a figure showing the average bone mass of each group at the age of 20 weeks. In the teriparatide administration test using an aging promoting model mouse (SAMP8), it is a figure showing the results of bone mass measurement by 2D-CT of representative individuals in each group at the age of 30 weeks. In a teriparatide administration test using an aging promoting model mouse (SAMP8), it is a figure showing the average bone mass of each group at the age of 30 weeks. In a teriparatide administration test using an aging promoting model mouse (SAMP8), it is a figure showing the average grip strength of each group at the age of 20 weeks. In a teriparatide administration test using an aging promoting model mouse (SAMP8), it is a figure showing the average grip strength of each group at the age of 30 weeks. In a teriparatide administration test using an aging promoting model mouse (SAMP8), the results of observation of the fast and slow muscles of the soleus muscle of representative individuals in each group at the age of 30 weeks with immunostained specimens with anti-SERCA1 antibody FIG. In the teriparatide administration test using an aging promotion model mouse (SAMP8), it is a figure which shows the result of having analyzed the UCP-3 expression level in the muscle of each group at the age of 30 weeks by Western blotting. In the teriparatide administration test using an aging promotion model mouse (SAMP8), it is a figure which shows the result of having analyzed the expression level of PGC-1 (alpha) in the muscle of each group at the age of 30 weeks by Western blotting. In the teriparatide administration test using senescence accelerated model mouse (SAMP8), it is a figure which shows the result of having analyzed the TNF- (alpha) expression level in the muscle of each group at the age of 30 weeks by Western blotting.

The present invention provides a pharmaceutical composition for preventing muscle aging, which contains parathyroid hormone (hereinafter referred to as “PTH”), a parathyroid hormone derivative (hereinafter referred to as “PTH derivative”) or a salt thereof as an active ingredient.
The origin of PTH used as an active ingredient in the present invention is not particularly limited, and PTH derived from various organisms can be used. Preferred is PTH derived from a mammal. Examples of mammals include, but are not limited to, humans, mice, rats, cows, pigs and the like. Among these, it is particularly preferable to use human PTH. As human PTH, natural human PTH, synthetic human PTH, recombinant human PTH and the like can be suitably used. Human PTH has the amino acid sequence shown in SEQ ID NO: 1.

PTH derivatives used as active ingredients in the present invention include PTH fragments, mutations in which 1 to several amino acids are deleted, substituted or added in the amino acid sequence of wild-type PTH, but exhibit biological activity of PTH And mutants in which 1 to several amino acids are conservatively substituted in the amino acid sequence of wild-type PTH. Preferably it is a variant of human PTH. As a variant of human PTH, a fragment of human PTH, a variant in which 1 to several amino acids are deleted, substituted or added in the amino acid sequence of human PTH (SEQ ID NO: 1), but exhibiting the biological activity of PTH, Examples include a variant in which one to several amino acids are conservatively substituted in the amino acid sequence of human PTH (SEQ ID NO: 1).

The human PTH fragment may be any fragment as long as it exhibits the biological activity of PTH, but is preferably a fragment containing 34 amino acids on the N-terminal side of human PTH, more preferably the N-terminal of human PTH. It is a fragment (teriparatide) consisting of 34 amino acids (SEQ ID NO: 2) on the side. As long as the fragment of human PTH shows the biological activity of PTH, the amino acid may be conservatively substituted.

PTH derivatives include those having a modified carboxy terminus. Examples of the modified carboxy terminus include carboxylate (—COO ⁻ ), amide (—CONH ₂ ), ester (—COOR) and the like. R in the ester is, for example, a C1-6 alkyl group such as methyl, ethyl, n-propyl, isopropyl or n-butyl, for example, a C3-8 cycloalkyl group such as cyclopentyl, cyclohexyl, etc., for example, phenyl, α-naphthyl, etc. In addition to C6-12 aryl groups such as benzyl, phenethyl, etc., C7-14 aralkyl groups such as phenyl-C1-2 alkyl groups such as benzyl and phenethyl or α-naphthyl-C1-2 alkyl groups such as α-naphthylmethyl, and the like Examples include pivaloyloxymethyl group, which is widely used as an ester.

PTH derivatives include those in which the side chain of the constituent amino acid is modified with an arbitrary substituent. Although a substituent is not specifically limited, For example, a fluorine atom, a chlorine atom, a cyano group, a hydroxyl group, a nitro group, an alkyl group, a cycloalkyl group, an alkoxy group, an amino group etc. are mentioned. In addition, substituents on the side chain of amino acids in the molecule (for example, —OH, —SH, amino group, imidazole group, indole group, guanidino group, etc.) are suitable protecting groups (for example, formyl group, acetyl, etc. C _And those protected with a C _1-6 acyl group such as a _2-6 alkanoyl group.

The PTH or PTH derivative used as an active ingredient in the present invention may form a salt. Examples thereof include salts with inorganic acids such as hydrochloric acid and sulfuric acid, and salts with organic acids such as acetic acid, tartaric acid, succinic acid, and malic acid. Acetate is preferred, and teriparatide acetate (acetate teriparatide) is particularly preferred.

A PTH or PTH derivative is expressed as a recombinant protein by constructing a recombinant expression vector into which a gene encoding PTH or a fragment thereof is inserted in a suitable host cell, for example, by a known genetic engineering technique. And can be produced by purification. The base sequence of the gene encoding PTH can be obtained from a known data base (DDBJ / GenBank / EMBL, etc.). As the base sequence of the gene encoding human PTH, for example, the base sequence shown in SEQ ID NO: 3 (ACCESSION: NM_000315) can be used.
Moreover, PTH or a PTH derivative can be manufactured using, for example, an in vitro transcription / translation system. PTH or a PTH derivative can be produced by a solid phase synthesis method (Fmoc method, Boc method) or a liquid phase synthesis method according to a known general peptide synthesis protocol.

The pharmaceutical composition of the present invention can effectively prevent muscle aging. In particular, since a decrease in muscle strength accompanying aging can be remarkably suppressed, it is very useful as a pharmaceutical for preventing and / or treating sarcopenia. Moreover, the pharmaceutical composition of the present invention is very useful as a medicament for preventing and / or treating locomotive syndrome. The muscle that is subject to aging prevention is not particularly limited, but skeletal muscle is preferred. Examples of skeletal muscles include the sternocleidomastoid muscle, the great pectoral muscle, the small pectoral muscle, the anterior sawnous muscle, the subclavian muscle, the rectus abdominis muscle, the external oblique muscle, the internal oblique muscle, the transverse abdominal muscle, the lumbar muscle, the monk. Cap muscle, latissimus dorsi, spine standing muscle, levator scapula, rhomboid, triangular muscle, small circular muscle, supraspinatus, subspinous muscle, subscapular muscle, great circular muscle, incisor arm, biceps , Brachial muscles, brachial muscles, triceps, elbows, circular rotator muscles, rectangular rotator muscles, supination muscles, ulnar carpal flexors, radial carpal flexors, long palmar muscles, superficial finger flexors, deep Finger flexor, long thumb flexor, long lateral carpal extensor, short lateral carpal extensor, ulnar carpal extensor, finger extensor, index finger extensor, little finger extensor, long thumb extensor, short thumb extensor Muscles, long thumb abductors, medial muscles (4 muscles), palmar interosseous muscles (3 muscles), dorsal interosseous muscles (4 muscles), little finger abductor, short finger flexor, little finger allele, Short palmar muscle, thumb adductor, short flexor flexor, thumb allele, short finger abductor, rectus femoris, lateral common, intermediate common, medial broad , Iliac muscle, psoas muscle, psoas muscle, sewing muscle, pubic muscle, thigh fascia latae muscle, gluteal muscle, biceps femoris, semi-tendonoid muscle, semi-membranous muscle, mid gluteus Muscle, thin muscle, long adductor, short adductor, major adductor, deep external rotation 6 muscles, gastrocnemius, soleus, popliteal muscle, posterior tibialis, long flexor, long mother flexor, sole Examples include muscle, anterior tibial muscle, long peroneal muscle, short peroneal muscle, third peroneal muscle, long mother's extensor muscle, and long peroneal extensor muscle. Preferred are the soleus, rectus femoris, and spinal column erect muscles.

The pharmaceutical composition of the present invention can be formulated by appropriately blending a pharmaceutically acceptable carrier and an additive with PTH, a PTH derivative or a salt thereof. Specifically, oral preparations such as tablets, coated tablets, pills, powders, granules, capsules, solutions, suspensions, emulsions; parenterals such as injections, infusions, suppositories, ointments, patches, etc. can do. What is necessary is just to set suitably about the mixture ratio of a carrier or an additive based on the range normally employ | adopted in the pharmaceutical field | area. Carriers or additives that can be blended are not particularly limited. For example, various carriers such as water, physiological saline, other aqueous solvents, aqueous or oily bases; excipients, binders, pH adjusters, disintegrants, absorption Various additives such as an accelerator, a lubricant, a colorant, a corrigent, and a fragrance are included.

Additives that can be mixed into tablets, capsules and the like include binders such as gelatin, corn starch, tragacanth, gum arabic, excipients such as crystalline cellulose, corn starch, gelatin, alginic acid and the like. Leavening agents, lubricants such as magnesium stearate, sweeteners such as sucrose, lactose or saccharin, flavorings such as peppermint, red oil and cherry. When the dispensing unit form is a capsule, a liquid carrier such as fats and oils can be further contained in the above type of material. Sterile compositions for injection can be prepared according to normal pharmaceutical practice (for example, dissolving or suspending an active ingredient in a solvent such as water for injection or natural vegetable oil). As an aqueous liquid for injection, for example, isotonic solutions containing physiological saline, glucose and other adjuvants (for example, D-sorbitol, D-mannitol, sodium chloride, etc.) are used. For example, alcohol (eg, ethanol), polyalcohol (eg, propylene glycol, polyethylene glycol), nonionic surfactant (eg, polysorbate 80 ^™ , HCO-50) and the like may be used in combination. As the oily liquid, for example, sesame oil, soybean oil and the like are used, and they may be used in combination with solubilizing agents such as benzyl benzoate and benzyl alcohol. Buffers (eg, phosphate buffer, sodium acetate buffer), soothing agents (eg, benzalkonium chloride, procaine, etc.), stabilizers (eg, human serum albumin, polyethylene glycol, etc.), storage You may mix | blend with an agent (for example, benzyl alcohol, phenol, etc.), antioxidant, etc.

Since the active ingredient of the pharmaceutical composition of the present invention is widely used for the treatment of osteoporosis, the pharmaceutical composition of the present invention can be used in humans and other mammals (for example, rats, mice, rabbits, sheep, pigs, cows, cats). , Dogs, monkeys, etc.).
The daily dose of the pharmaceutical composition of the present invention is not particularly limited as long as it is an amount effective for preventing muscle aging and has few side effects. The daily dose of the pharmaceutical composition of the present invention is preferably set according to the daily dose of a commercially available osteoporosis therapeutic agent.

Hereinafter, the present invention will be described in detail by way of examples, but the present invention is not limited thereto.

[Example 1: Effect of teriparatide on muscle strength of senescence accelerated model mice]
(1) Animals Used SAMP8 (7 weeks old, male) which is an aging promoting model mouse and SAMR1 (7 weeks old, male) which is a normal aging mouse were obtained from SLC Japan.

(2) Experimental method (2-1) Breeding and administration The experimental protocol and breeding conditions were approved by the Osaka University Animal Experiment Committee, and the experiment was conducted in accordance with the “National Institute of Health Research and Guide for Laboratory Animal Care and Use”. Carried out.
Tested after one week of habituation. The animals were reared in a specific pathogen-free breeding room with a 12-hour light-dark cycle, and were allowed to freely take normal food (MF, Oriental Yeast Co., Ltd.) and tap water. SAMP8 and SAMR1 were divided into two groups, a teriparatide administration group (n = 5) and a solvent administration group (n = 5), respectively, and were raised to 20 or 30 weeks of age. The general condition of all mice was observed once every 2 weeks during the breeding period.

(2-2) Preparation of specimen A mouse serum albumin dissolved in physiological saline to a final concentration of 0.1% was used as a solvent, and 40 micrograms of teriparatide dissolved in 1 ml of solvent was used as a teriparatide administration liquid. Specifically, 100 mg / vial of mouse serum albumin was dissolved in 100 ml of physiological saline in a safety cabinet to prepare 0.1% mouse serum albumin physiological saline. Except for dissolving teriparatide, approximately 1.5 ml was dispensed for administration, frozen at −30 ° C., and thawed before use. The teriparatide dosing solution was prepared by dissolving 0.5 mg / vial teriparatide in 37.5 ml of solvent in a safety cabinet to prepare 1.2 μg / 90 μl teriparatide dosing solution. About 1.5 ml was dispensed, frozen at −30 ° C. and thawed before use.

(2-3) Administration Body weight was measured every Monday, and teriparatide 40 μg / kg B.W. (0.4 μg / 10 g B.W.) or solvent was administered subcutaneously on Monday, Wednesday and Friday based on the body weight. . The dose volume per animal was 30 μl / 10 g BW.

(2-4) Sample collection After weighing at 20 and 30 weeks of age, the mice were euthanized, and the lower limbs (including the femur and tibia) were removed. The femur and tibia were subjected to tissue preparation. In addition, the soleus and rectus femoris were removed, immediately frozen in liquid nitrogen, and then stored at −80 ° C. Cryopreserved soleus and rectus femoris were thawed at a later date, homogenized, and subjected to Western blotting. In addition, the paravertebral column standing muscle was excised, the thoracic vertebrae to the coccyx were removed, and the bone mass of the first lumbar vertebra was measured.

(2-5) Bone Mass Measurement 2D bone measurement TRI / 2D-BON (Ratok System Engineering Co., Ltd., Tokyo) was used. Using the cut lumbar spine, cancellous bone and cortical bone were taken out from the designated area according to the manual, the measurement range was designated, and 2D trabecular structure measurement was performed.

(2-6) Measurement of grip strength The grip strength of a mouse was measured using a small animal grip strength measuring device GPM-100B (Merquest, Toyama City). Place the mouse's forelimb heel, display, and middle heel on the net, confirm that it is firmly fixed, then pull the mouse horizontally to the rear and apply force until the mouse releases the net. The maximum grip strength at the moment of separation was measured, and the unit was expressed in grams.

(2-7) Tissue specimen In order to observe the fast and slow muscles in the soleus, tissue specimens of the tibia were prepared according to a conventional method. Using anti-SERCA1 antibody (Anti-SERCA1 ATPase Rabbit monoclonal Ab, Abcam (ab133275)) using SERCA1 (fast-twitch skeletal muscle sarcoplasmic reticulum Ca (2+) ATPase) as an index went.

(2-8) Analysis of UCP-3 expression level in muscle by Western blotting Cryopreserved muscle was homogenized with RIPA buffer supplemented with a protease inhibitor cocktail (Nacalai Tesque). Centrifugation was performed at 14,000 rpm for 5 minutes at 4 ° C, and the supernatant was collected. Protein quantification was performed using a BCA protein assay kit (Thermo Fisher Scientific Inc.). An equal amount of protein was used, heated at 95 ° C. for 5 minutes, and subjected to SDS-PAGE. For SDS-PAGE, a 15% polyacrylamide gel was used. After transfer to the PVDF membrane, anti-UCP-3 antibody (Anti-UCP-3 Rabbit polyclonal Ab, abcam (ab3477)) was diluted 1000 times and incubated at 4 ° C. for 16 hours, followed by horseradish peroxidase labeled secondary antibody (GE (Healthcare Bio-Sciences Corp.) was developed using Immunostar Zeta (Wako Pure Chemical Industries), photographed with RX-U film (Fuji Film), captured with GT-X970, and evaluated.

(2-9) Analysis of expression level of PGC-1α in muscle by Western blotting Using a 20-20% gradient polyacrylamide gel for SDS-PAGE, anti-PGC-1α antibody (Anti-PGC-1α Mouse mAb ( 4C1.3), Calbiochem (ST1202)) is diluted 1000-fold, and anti-GAPDH antibody (GAPDH (14C10) Rabbit monoclonal Ab (Cell Signaling Technology)) is diluted 10000-fold. Western blotting was performed as in 8).

(2-10) Analysis of TNF-α expression level in muscle by Western blotting Using a 20-20% gradient polyacrylamide gel for SDS-PAGE, anti-TNF-α antibody (TNF-α rabbit polyclonal Ab (CST (# 3707)) was diluted 500-fold and anti-GAPDH antibody (GAPDH (14C10) Rabbit monoclonal Ab (Cell Signaling Technology)) was diluted 10000-fold in the same manner as (2-8) above. Western blotting was performed.

(3) Results (3-1) Body weight The average body weight of each group at the age of 20 weeks is shown in FIG. 1, and the average body weight of each group at the age of 30 weeks is shown in FIG.
1 and 2, P8 represents a SAMP8 solvent administration group, P8 + T represents a SAMP8 teriparatide administration group, R1 represents a SAMR1 solvent administration group, and R1 + T represents a SAMR1 teriparatide administration group (FIGS. 4, 6, 7 and 8 are also shown). the same). At 20 and 30 weeks of age, the average body weight of SAMP8 was lower than the average body weight of SAMR1. At 20 weeks of age, there was no significant difference between the solvent-administered group and the teriparatide-administered group for both SAMP8 and SAMR1 (Welch test). At 30 weeks of age, in SAMP8, the average body weight was significantly higher in the teriparatide administration group than in the solvent administration group (Welch test, p = 0.0377).

(3-2) Bone Mass The results of 2D-CT of representative individuals in each group at the age of 20 weeks are shown in FIG. 3, and the average bone mass in each group is shown in FIG. The results of 2D-CT of representative individuals in each group at 30 weeks of age are shown in FIG. 5, and the average bone mass of each group is shown in FIG. 3 and 5, P8 represents SAMP8 and R1 represents SAMR1. At 20 weeks of age, there was no significant difference in average bone mass between SAMP8 and SAMR1, and neither SAMP8 nor SAMR1 was significant between the solvent-administered group and the teriparatide-administered group (Welch's test). ). Even at 30 weeks of age, no significant difference in average bone mass was observed between SAMP8 and SAMR1. On the other hand, there was no statistically significant difference at the age of 30 weeks, but both TAMP8 and SAMR1 tended to have higher average bone mass in the teriparatide administration group than in the solvent administration group. That is, in any mouse, an increase in bone mass based on the original pharmacological action of teriparatide was confirmed.

(3-3) Muscle Strength The average grip strength of each group at the age of 20 weeks is shown in FIG. 7, and the average grip strength of each group at the age of 30 weeks is shown in FIG. Both SAMP8 and SAMR1 showed a tendency for grip strength at 30 weeks of age to be lower than at 20 weeks of age. At the age of 20 weeks, no significant difference was observed between the solvent administration group and the teriparatide administration group (Welch's test). At 30 weeks of age, the average grip strength of the teriparatide-administered group was significantly higher than that of the solvent-administered group in the aging-promoted model mouse SAMP8 (Welch test, p = 0.0397). On the other hand, in SAMR1 of normal aging mice, no significant difference was observed between the solvent administration group and the teriparatide administration group (Welch's test, p = 0.4526).

(3-4) Analysis of fast and slow muscles In sarcopenia, it is known that fast muscles decrease and slow muscles increase. Therefore, the fast muscle (SERCA1 positive) and the slow muscle (SERCA1 negative) of the soleus muscle specimen were observed by immunostaining with anti-SERCA1 antibody. The results are shown in FIG. In the figure, the light gray part is the fast muscle and the whitish part is the whitish muscle. In the SAMP8 solvent administration group, the amount of slow muscle was increased, but in the SAMP8 teriparatide administration group, the amount of slow muscle was reduced, which was equivalent to SAMR1.

(3-5) Analysis of UCP-3 expression level in muscle UCP (Uncoupling protein) is a mitochondrial inner membrane protein, and UCP-3 is often present in muscle tissues such as skeletal muscle. Are known. Therefore, the expression level of UCP-3 in the muscle of each group of mice was analyzed by Western blotting. The results are shown in FIG. In the SAMP8 solvent administration group, the expression level of UCP-3 was decreased, but in the SAMP8 teriparatide administration group, the decrease in the expression level was suppressed.

(3-6) Analysis of expression level of PGC-1α in muscle It is known that the strength of muscle fibers depends on the expression level of PGC-1α in the physiological range (reference: Lin J et al. Transcriptional co-activator PGC-1 alpha drives the formation of slow-twitch muscle fibers. Nature. 2002 Aug 15; 418 (6899): 797-801.). Therefore, the expression level of PGC-1α in the muscle of each group of mice was analyzed by Western blotting. The results are shown in FIG. The expression level of PGC-1α was decreased in the SAMP8 solvent administration group, but the expression level of PGC-1α was recovered in the SAMP8 teriparatide administration group, which was equivalent to SAMR1.

(3-7) Analysis of expression level of TNF-α in muscle The expression level of TNF-α in the muscle of each group of mice was analyzed by Western blotting. The results are shown in FIG. In the SAMP8 solvent administration group, the expression level of TNF-α was higher than that in SAMR1, and it was considered that inflammation occurred in the muscle. However, in the SAMP8 teriparatide administration group, the expression level of TNF-α was suppressed.

(4) Discussion As described above, when teriparatide was administered, increase in bone mass based on the original pharmacological action was confirmed in both SAMP8 of senescence-accelerating model mice and SAMR1 of normal aging mice. Was observed only in a model mouse for aging promotion (SAMP8). Moreover, by administering teriparatide to an aging promoting model mouse (SAMP8), the fast muscle increased and the balance between the fast muscle and the slow muscle was improved. As the molecular mechanism, the expression level of UCP-3 and the expression level of PGC-1α were also increased by the administration of teriparatide. Therefore, the mitochondrial function via UCP-3 is considered to be restored. It was. Furthermore, since the expression level of TNF-α in the muscle of the aging promoting model mouse (SAMP8) was suppressed by administration of teriparatide, it was considered that the inflammation of the muscle due to aging was suppressed.
The present inventors have already shown that an aging-promoting model mouse (SAMP8) is suitable as a sarcopenia model (The 55th Annual Meeting of the Japan Geriatrics Society, General Presentation O-4, May 2013) Issued on the 7th of May). That is, from this result, it was considered that sarcopenia can be prevented or treated by teriparatide administration.

The present invention is not limited to the above-described embodiments and examples, and various modifications are possible within the scope shown in the claims, and technical means disclosed in different embodiments are appropriately combined. The obtained embodiment is also included in the technical scope of the present invention. Moreover, all the academic literatures and patent literatures described in this specification are incorporated herein by reference.

Claims

A pharmaceutical composition for preventing muscle aging, comprising parathyroid hormone, a parathyroid hormone derivative or a salt thereof as an active ingredient.
The pharmaceutical composition according to claim 1, wherein the parathyroid hormone derivative is teriparatide.
3. The pharmaceutical composition according to claim 1 or 2, which is used for suppressing muscle weakness.
The pharmaceutical composition according to any one of claims 1 to 3, which is used for prevention and / or treatment of sarcopenia.
The pharmaceutical composition according to any one of claims 1 to 3, which is used for prevention and / or treatment of locomotive syndrome.