KR101919081B1 - Composition for prevention or treatment of muscular atrophy or sarcopenia and improvement of muscular functions comprising human placenta extract - Google Patents
Composition for prevention or treatment of muscular atrophy or sarcopenia and improvement of muscular functions comprising human placenta extract Download PDFInfo
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- KR101919081B1 KR101919081B1 KR1020170058863A KR20170058863A KR101919081B1 KR 101919081 B1 KR101919081 B1 KR 101919081B1 KR 1020170058863 A KR1020170058863 A KR 1020170058863A KR 20170058863 A KR20170058863 A KR 20170058863A KR 101919081 B1 KR101919081 B1 KR 101919081B1
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- placenta extract
- muscle
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- human placenta
- muscular
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- A—HUMAN NECESSITIES
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Abstract
Description
본 발명은 인간 태반 추출물을 유효성분으로 포함하는 근 위축증 또는 근육감소증의 예방 또는 치료용 및 근육 기능 개선용 조성물에 관한 것이다.The present invention relates to a composition for preventing or treating muscular dystrophy or muscular dysgenesis comprising human placenta extract as an active ingredient and a composition for improving muscle function.
근육을 형성하는 근육세포는 근필라멘트로 구성되어 있는데, 이는 세포의 크기를 변하도록 하여 수축을 유발한다. 근육 조직이 수축할 때 장력이 발생하게 되는데, 이를 근력이라고 한다. 근육은 이러한 근력을 통하여 개체의 이동, 자세 유지, 체액분비 등을 담당하는 신체기관이다.Muscle cells that form muscles are composed of muscle filaments, which cause the size of the cells to change causing shrinkage. When muscle tissue contracts, tension is generated, which is called muscle strength. Muscles are the body organs responsible for movement, posture, and secretion of fluid through these muscles.
근육질환은 점진적인 근력감소로 인해 발생하는 진행성 질환으로, 대개 염색체 이상으로 발생하며 돌연변이, 염증, 신진대사의 이상, 내분비 기능의 장애, 독성 등 다양한 원인이 있다. 이러한 원인들로 인해 발생하는 근육 질환으로는 근이영양증(muscular dystrophy), 근육긴장증(myotonia), 근 위축증(muscular atrophy), 근육감소증(sarcopenia)등이 있다.Muscle disease is a progressive disease caused by gradual decrease in muscle strength, usually occurs as a chromosome abnormality, and has various causes such as mutation, inflammation, abnormal metabolism, impaired endocrine function, and toxicity. Muscular diseases caused by these causes include muscular dystrophy, myotonia, muscular atrophy, and sarcopenia.
한편, 인간 태반 추출물(Human placenta extract, HPE)은 다양한 성장인자, 사이토카인 및 기타 생리활성물질을 포함하며, 피로의 경감, 항산화 등의 용도로 널리 이용되고 있다(Lee KK, et al., Evid Based Complement Alternat . Med ., vol. 2012,(2012) p.130875). 또한, 인간 태반 추출물은 동물을 이용한 연구에서 간 재생을 통해 간 기능 향상을 유도하고, TGF-β 및 VEGF를 생산함으로써 상처 치료에 영향을 미친다고 알려져 있다. 그러나, 인간 태반 추출물에 대한 많은 관심에도 불구하고 인간 태반 추출물의 기능에 대해서는 아직 완전히 연구되지 않았으며, 특히 근 질환에 대한 연구는 진행된 바 없다.Meanwhile, human placenta extract (HPE) contains various growth factors, cytokines and other physiologically active substances, and is widely used for relieving fatigue and antioxidation (Lee KK, et al ., Evid Based Complement Alternat . Med ., Vol. 2012, (2012) p.130875). In addition, human placenta extracts are known to induce liver function improvement through liver regeneration in animal studies and to affect wound healing by producing TGF-β and VEGF. However, despite the great interest in human placenta extract, the function of human placenta extract has not yet been thoroughly studied, and studies of muscle disorders have not been conducted.
이에, 본 발명자는 근육 질환을 예방 또는 치료할 수 있는 물질을 연구, 개발하기 위하여 노력하던 중, 인간 태반 추출물을 C2C12 근아세포(myoblast cell)에 처리함에 따라 산화스트레스에 대한 세포 보호작용을 확인함으로써 본 발명을 완성하였다.Accordingly, the inventors of the present invention have made efforts to research and develop substances capable of preventing or treating muscle diseases, and have found that by treating human placenta extract with C2C12 myoblast cells, Thereby completing the invention.
본 발명의 목적은 태반 추출물을 유효성분으로 포함하는 근육 질환 예방 또는 치료용 조성물을 제공하는 것이다.It is an object of the present invention to provide a composition for preventing or treating a muscle disorder comprising placenta extract as an active ingredient.
본 발명의 다른 목적은 태반 추출물을 유효성분으로 포함하는 근육 기능 개선용 조성물을 제공하는 것이다.Another object of the present invention is to provide a composition for improving muscle function comprising placenta extract as an active ingredient.
상기 목적을 달성하기 위해, 본 발명은 태반 추출물을 유효성분으로 포함하는 근 위축증(muscular atrophy) 또는 근육감소증(sarcopenia)의 예방 또는 치료용 약학적 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating muscular atrophy or sarcopenia comprising placenta extract as an active ingredient.
상기 다른 목적을 달성하기 위해, 본 발명은 태반 추출물을 유효성분으로 포함하는 근육 기능 개선용 약학적 조성물 또는 식품 조성물을 제공한다.To achieve these and other objects, the present invention provides a pharmaceutical composition or food composition for improving muscle function comprising placenta extract as an active ingredient.
본 발명에 따른 조성물은 산화스트레스 상황에 있는 근육세포의 증식을 증가시키고 세포 내 활성산소량을 감소시키며, 미토콘드리아 합성의 마스터 단백질인 PGC1-alpha의 발현량을 증가시켜 미토콘드리아 합성을 증가시킬 수 있다. 또한, 본 발명의 조성물은 오토파지를 활성화 시키고, 근 위축 모델에서 근 볼륨을 증가시킴으로써 근 위축증 또는 근육감소증의 예방 또는 치료, 및 근육 기능 개선에 유용하게 활용될 수 있다.The composition according to the present invention can increase the proliferation of muscle cells in the oxidative stress state, decrease the intracellular active oxygen amount, and increase the expression of PGC1-alpha, the master protein of mitochondrial synthesis, thereby increasing mitochondrial synthesis. In addition, the composition of the present invention can be usefully used for preventing or treating muscular dystrophy or muscular dysfunction, and improving muscle function by activating autophagy and increasing muscle volume in an atrophy model.
도 1a는 H2O2 500 uM을 최고 농도로 정하여 1/2 희석으로 최종 128배 희석한 농도까지 처리한 후, 24시간 뒤 CCK-8 solution을 이용하여 세포 생존율을 확인한 그래프이다.
도 1b는 라이넥 10%를 최고농도로 정하여 1/2 희석으로 최종 128배 희석한 농도까지 처리한 후, 24시간 뒤 세포 생존율을 확인한 그래프이다.
도 1c는 라이넥(5%) 및 H2O2(250 uM) 처리 후, 4시간 뒤 세포 생존율을 나타낸 그래프이다.
도 1d는 라이넥(5%) 및 H2O2(250 uM) 처리 후, ROS probe (H2-DCF, DCF/thermo/ D339/15 uM)를 사용하여 세포질의 ROS 염색을 통해 microplate reader기를 이용하여 형광량을 측정한 것을 나타낸 그래프이다.
도 2a는 C2C12 세포에 라이넥(5%) 및 H2O2(250 uM) 처리 후, 세포의 용해물을 확보하여 웨스턴블롯을 이용하여 단백질 발현량을 비교한 것을 나타낸 것이다.
도 2b는 C2C12 세포에 라이넥(5%) 및 H2O2(250 uM) 처리 후, mito-tracker를 이용하여 미토콘드리아를 염색하고, 마이크로 플레이트를 이용하여 형광량을 측정하여 대조군(Con) 대비 상대값을 나타낸 것이다.
도 3은 C2C12 세포에 라이넥(5%) 및 H2O2(250 uM) 처리 후, 세포의 용해물을 확보하여 웨스턴블롯을 통해 단백질 발현량을 비교하여 나타낸 것이다.
도 4a 및 도 4b는 EMG 모니터링 장치를 이용하여 Normal, 보톡스(BTX) 처리군 및 BTX와 라이넥 처리군, 총 3개 집단의 근전도 변화를 관찰 초기 및 마지막 날짜에 확인한 그래프이다.
도 5a는 대조군, BTX 처리군, BTX 및 라이넥 처리군의 마우스를 희생시킨 다음 우측 뒷다리의 비복근을 실체현미경(Stereoscopic microscope)을 이용하여 육안으로 평가한 것이다.
도 5b는 PRIMOSLITE 소프트웨어(PRIMOSLITE Vesrsion 5.8E)를 이용하여 대조군, BTX 처리군, BTX 및 라이넥 처리군에 속하는 마우스의 우측 뒷다리 비복근의 질량을 분석하여 나타낸 것이다.
도 5c는 각 집단의 마우스의 우측 다리 비복근을 PRIMOSLITE를 이용하여 스캔한 후, 볼륨을 측정하여 값을 그래프로 나타낸 것이다. 이때, 결과값은 마우스 한 마리 당 3번 측정한 값의 평균이다.FIG. 1A is a graph showing the cell survival rate using a CCK-8 solution after 24 h after treatment with 500 μM of H 2 O 2 at the highest concentration and diluting to a final dilution of 128 times with 1/2 dilution.
FIG. 1B is a graph showing the cell survival rate after 24 hours after treatment with a final concentration of 10% linac to a final dilution of 128 by 1/2 dilution.
Figure 1c is a graph showing cell viability after 4 hours of treatment with lane (5%) and H 2 O 2 (250 uM).
Utilization 1d is Lai neck (5%) and H 2 O 2 (250 uM) an microplate reader with a ROS staining of the cytoplasm using, ROS probe (H2-DCF, DCF / thermo / D339 / 15 uM) after treatment And the amount of mold light is measured.
Figure 2a shows that the securing to the Rai-neck (5%) and H 2 O 2 (250 uM) After treatment, cell lysates of the C2C12 cells using Western blot comparison of protein expression levels.
FIG. 2b shows the results of treatment of C2C12 cells with lignect (5%) and H 2 O 2 (250 μM), followed by staining mitochondria using a mito-tracker and measuring the amount of fluorescence using a microplate. Relative value.
Figure 3 compares the amount of protein expression by western blot securing cell lysates after treatment with lanectin (5%) and H 2 O 2 (250 uM) on C2C12 cells.
FIGS. 4A and 4B are graphs showing EMG changes in three groups, Normal, Botox (BTX), BTX and the lyne treatment group, using an EMG monitoring device at the initial and final dates of observation.
FIG. 5A is a visual evaluation of the gastrocnemius muscle of the right hind leg using a stereoscopic microscope after sacrificing the mice of the control group, the BTX treatment group, the BTX and the lynek treatment group.
FIG. 5B shows the analysis of the mass of the right hind gland gastrocnemius of mice belonging to the control group, the BTX treatment group, the BTX treatment, and the linac treatment group using the PRIMOS LITE software (PRIMOS LITE Vession 5.8E).
FIG. 5C is a graph showing values obtained by scanning the right leg gait of each group of mice using a PRIMOS LITE and measuring the volume. At this time, the result is the average of three measurements per mouse.
본 발명은 일 측면으로, 태반 추출물을 유효성분으로 포함하는 근 위축증(muscular atrophy) 또는 근육감소증(sarcopenia)의 예방 또는 치료용 약학적 조성물을 제공한다.In one aspect, the present invention provides a pharmaceutical composition for preventing or treating muscular atrophy or sarcopenia comprising placenta extract as an active ingredient.
본 발명에서 "태반 추출물(placenta extract)"은 인간 또는 동물의 태반에서 수득된 추출물이다. 상기 추출물은 태반에 산 및/또는 효소를 처리하여 수득될 수 있다. 이때, 동물은 포유동물, 구체적으로 인간, 소, 말, 양, 또는 돼지 등의 유래일 수 있다.In the present invention, "placenta extract" is an extract obtained in human or animal placenta. The extract can be obtained by treating the placenta with an acid and / or an enzyme. Here, the animal may be derived from a mammal, particularly a human, cow, horse, sheep, or pig.
상기 "태반"은 임신과 함께 생성되는 장기로서, 모체에서 태아로 영양물과 효소의 공급, 태아에서 생기는 노폐물과 탄산가스 등을 모체로 배출, 생체에 있어서 이물질인 병원균이나 약물의 태아로의 이행저지 및 태아의 내분비조정 등의 기능을 한다.The term " placenta " is a organs produced with the pregnancy, and it is a organ produced by feeding the nutrients and enzymes from the mother to the fetus, exhausting the waste materials and carbon dioxide generated in the fetus to the mother, inhibiting the foreign body, And endocrine regulation of the fetus.
태반에는 아미노산류, 단백질류, 당류, 핵산류, 지질류, 무기물, 효소류, 호르몬류 등을 함유한다. 이때, 아미노산류는 아스파라긴산, 글루타민산, 루이신, 라이신, 글리신, 알라닌, 세린, 스레오닌, 페닐알라닌, 타이로신, 메치오닌, 히스티딘 등이 있다. 단백질 및 효소류는 알부민, 글로불린, 산성 및 알칼리성 포스파타제, 하알로니다제 등을 포함한다. 당류는 글루코스, 갈락토스, 리보스 등을 포함한다. 핵산류는 우라실, 크산틴, 히포크산틴 등을 포함하며, 지질류는 라우린산, 팔미틸산, 리놀레인산 등이 있다. 무기물은 Na, K, Ca, P, Fe, Cl 등을 포함하며, 호르몬류에는 고나도트롬빈, 락토겐, 스테로이드 호르몬 등이 있다.The placenta contains amino acids, proteins, sugars, nucleic acids, lipids, minerals, enzymes, hormones and the like. At this time, the amino acids include aspartic acid, glutamic acid, leucine, lysine, glycine, alanine, serine, threonine, phenylalanine, tyrosine, methionine and histidine. Proteins and enzymes include albumin, globulin, acidic and alkaline phosphatase, and allodynia. Saccharides include glucose, galactose, ribose, and the like. The nucleic acids include uracil, xanthine, hypoxanthin and the like, and the lipids include lauric acid, palmitic acid, and linoleic acid. Minerals include Na, K, Ca, P, Fe, and Cl, and hormones include gonadothrombin, lactogens, and steroid hormones.
본 발명의 일 실시예에서는, 인간 태반 추출물로서 녹십자웰빙에서 제조한 라이넥주(Laennec Inj.)를 사용하여 근 위축 모델에서 근 위축증 및 근육감소증에 대한 예방 및 치료 효과를 관찰하였다.In one embodiment of the present invention, prophylactic and therapeutic effects of muscular atrophy and muscle hypoxia were observed in an atrophy model using Laennec Inj. Manufactured by Green Cross Inc. as human placenta extract.
본 발명에서 용어 "근 위축증"이란 근육이 위축되는 질환으로서, 팔다리 근육이 좌우대칭적으로 점점 위축되어가는 질환이다. 근 위축증에는 여러 종류가 있으며, 가장 많이 발생하는 질환으로는 근 위축성 측색경화증(amyotrophic lateral sclerosis, ALS)과 척수성진행성 근 위축증(spinal progressive muscular atrophy)이 있다.In the present invention, the term " muscular atrophy " refers to a disease in which muscles are contracted, and limb muscles are gradually contracted symmetrically. There are many types of muscle atrophy, and the most common diseases are amyotrophic lateral sclerosis (ALS) and spinal progressive muscular atrophy.
근 위축성 측색경화증은 루게릭병(Lou Gehrig's disease)이라고도 불리우며, 뇌간과 척수에 있는 하운동신경원세포와 함께 대뇌피질과 상운동신경계를 침범하여 생기는 퇴행성 질환이다. 팔다리와 얼굴 주위의 근육이 점차적으로 마르고 힘이 없어지며, 팔다리를 움직일 때 뻣뻣해지는 증세가 나타난다. 그러나, 저리거나 아픈 감각 증상이 없으며, 의식이 명료하고, 안구운동 장애 또는 배변 및 배뇨 장애가 없다. 대부분 40-70세에 연간 인구 10만 명당 0.4 내지 2명 정도로 발병하며, 남성 환자 수가 여성 환자 수보다 약간 많고, 대부분 발병한지 2-5년 이내에 호흡마비로 목숨을 잃는다. 근 위축성 측색경화증은 가족력이 있을 수 있고, 이 외에 원인이 아직 밝혀져 있지 않다. 상기 질환을 예방 또는 치료할 수 있는 특별한 방법이 아직 없다.Amyotrophic lateral sclerosis, also called Lou Gehrig's disease, is a degenerative disease caused by invasion of the cerebral cortex and upper motor nervous system along with lower motor neurons in the brainstem and spinal cord. Muscles around the limbs and face slowly dry out, lose strength, and become stiff when moving the limbs. However, there is no numbness or sore sensory symptoms, clear consciousness, no eye movement disorder or defecation and dysuria. Most people age 40-70 have an annual outbreak of about 0.4 to 2 people per 100,000 population, and the number of male patients is slightly more than the number of female patients, and most of them die from respiratory paralysis within 2-5 years of their onset. Amyotrophic lateral sclerosis may have a family history, and the cause is unknown yet. There is no specific way to prevent or treat the disease.
척수성진행성 근 위축증은 척수 및 연수의 운동신경세포의 변성에 의하여 전신의 근 위축과 탈력을 일으키는 질환이다. 근육 위축이 손발에서 시작하여 점차 상행하여 목의 근육과 몸통의 근육이 침해된다. 상하지의 힘줄 반사가 약해지고 바빈스키 반사에는 음성이 나타난다. 3세 이후의 어느 연령층에서나 발병하며 긴 경과를 취하고, 조기에 사망하는 일이 없다. 그러나, 갓난 아기에서 상기 질병이 나타나는 것을 "베르트니히-호프만병"이라고 하며, 이 경우 수년 이내에 사망할 수 있다. 척수성진행성 근 위축증을 예방 또는 치료할 수 있는 특별한 방법이 아직 없는 실정이다.Spinal cord sinus rhythm muscular atrophy is a disease that causes muscle atrophy and degeneration of the whole body by degeneration of motor neurons of spinal cord and soft tissue. Muscle atrophy begins at limbs and progresses gradually to inflate muscles of the neck and body. The tendon reflexes of the upper and lower parts are weakened, and the voice of Babinsky is reflected. It occurs in any age group after 3 years, takes a long time, and does not die prematurely. However, the emergence of the disease in newborn babies is referred to as "Bertrnevich-Hoffmann disease", and in this case they can die within a few years. There is no specific way to prevent or treat spinal sinus rhythm muscular atrophy.
본 발명에서 용어 "근육감소증"이란 노화가 진행됨에 따라 근육의 밀도가 감소하고 기능이 점차적으로 약화되어 근육이 감소되는 것을 의미한다. 상기 질환은 노화로 인해 근육이 감소하여, 노인 근육감소증(age-related sarcopenia)라고도 한다. 근육감소증은 노화와 함께 신체활동이 적은 사람에게 발병하지만 이 외에도 뇌에서 몸이 움직이도록 근육에 신호를 보내는 신경 세포의 감소, 성장호르몬, 남성호르몬 등의 감소, 체내 단백질 합성 능력의 감소 등의 원인으로 인해 발병한다. 치료법으로는 근력운동, 약물치료, 호르몬 대체치료 등이 있으며, 호르몬 대체치료를 장기간 진행할 경우 암이나 그 외에 건강상의 문제를 일으킬 수 있다.The term " myopenia " in the present invention means that as the aging progresses, the density of the muscle decreases and the function gradually weakens and the muscle decreases. The disease is also referred to as age-related sarcopenia, as the muscle declines due to aging. Muscle hypoxia causes aging and low physical activity, but it also causes decrease in nerve cells that signal muscles to move the body in the brain, decrease of growth hormone, male hormone, etc. . Treatment includes strength training, medication, and hormone replacement therapy. Prolonged hormone replacement therapy can cause cancer and other health problems.
본 발명은 다른 측면으로, 태반 추출물을 유효성분으로 포함하는 근육 기능 개선용 약학적 조성물을 제공한다.In another aspect, the present invention provides a pharmaceutical composition for improving muscle function comprising placenta extract as an active ingredient.
상기 용어 "근육 기능"이란, 근육의 수축에 의해 힘을 발휘하는 능력을 의미하며, 근육이 저항을 이겨내기 위하여 최대한으로 수축력을 발휘할 수 있는 능력인 근력, 근육이 주어진 중량에 얼마나 오랫동안 또는 얼마나 여러 번 수축과 이완을 반복할 수 있는지를 나타내는 능력인 근지구력, 단시간 내에 강한 힘을 발휘하는 능력인 순발력을 포함한다. 이러한 근력 기능은 간이 주관하며, 근육량에 비례한다. 또한, 상기 용어 "근육 기능 개선"은 근육 기능을 더 좋게 향상시키는 것을 의미한다.The term " muscle function " means an ability to exert a force by contraction of a muscle, such as muscle strength, which is the ability of a muscle to exert its maximum retraction force to overcome resistance, Muscle endurance which is the ability to show whether it can repeat the contraction and relaxation, and the ability to act strongly within a short time. This muscle function is hosted by the liver and is proportional to the muscle mass. The term " improving muscle function " also means improving muscle function better.
상기 근육은 골격근육(skeletal muscle)일 수 있다. 골격근육은 뼈나 힘줄에 붙어서 수의적으로 수축하여 움직임이나 힘을 만드는 연부 조직이다. 근육은 수축만 가능해서 붙어 있는 힘줄이나 뼈를 서로 끌어 당기는 역할만 하지만 뼈에 붙어 있는 위치나 방향에 따라 다양한 동작이 가능하고, 수축하는 정도에 따라 다양한 강도의 힘을 낼 수 있다. 골격근육은 다른 근육과 달리 수의적으로 조절할 수 있어 우리가 원하는 동작을 할 수 있게 한다.The muscle may be a skeletal muscle. Skeletal muscle is a soft tissue that attaches to bones or tendons and contracts to move or force. Muscles are only able to contract, so they pull the attached tendons and bones together, but they can perform various actions depending on the position and direction attached to the bones, and can produce various strengths depending on the degree of contraction. Skeletal muscles, unlike other muscles, can be manipulated voluntarily, allowing us to perform the desired action.
본 발명의 일 실시예에서, 본 발명에 따른 태반 추출물인 라이넥의 농도가 증가할수록, 산화 스트레스 상황에서 근세포의 생존율이 증가함을 확인하였다(도 1b). 또한, 라이넥을 처리한 군에서, 처리하지 않은 군에 비해 근 볼륨이 크게 유지됨을 확인하였다(도 5a 내지 도 5c). 따라서, 본 발명에 따른 태반 추출물은 산화 스트레스 상황에 있는 근육세포의 증식 및 생존율을 증가시키고 세포 내 활성산소량을 감소시키며, 근 볼륨을 크게 증가시킴으로써 근 위축증 또는 근육감소증의 예방 또는 치료, 및 근육 기능 개선에 유용하게 사용될 수 있다.In one embodiment of the present invention, it was confirmed that the survival rate of the myocardial cells was increased as the concentration of the placenta extract, linecen, increased according to the oxidative stress (FIG. 1B). In addition, it was confirmed that the muscle volume was largely maintained in the group treated with the linecn compared to the group not treated (Fig. 5A to Fig. 5C). Therefore, the placenta extract according to the present invention can increase the proliferation and survival rate of muscle cells in the oxidative stress condition, decrease the intracellular active oxygen amount, and significantly increase the muscle volume to prevent or treat muscular dystrophy or myopinia, Can be usefully used for improvement.
또한, 본 발명에 따른 약학적 조성물의 제형은 주사제, 또는 피부 외용제의 형태, 예를 들면 주사제, 크림, 젤, 패치, 분무제, 연고제, 경고제, 로션제, 마이크로니들, 롤러, 및 이의 조합으로 이루어진 군에서 선택되는 형태일 수 있다.In addition, the pharmaceutical composition according to the present invention may be in the form of an injection or an external preparation for skin such as an injection, a cream, a gel, a patch, a spray, an ointment, an alarm, a lotion, a micro needle, a roller, And the like.
또한, 본 발명의 조성물은 치료 부위에 직접 투여하기에 적합한 피부 외용제 형태로 제제화될 수 있으며, 이 경우 피부에 국소 투여하기 적합한 생리학적으로 허용가능한 담체 또는 희석제 등을 추가로 포함할 수 있다. 상기 생리학적으로 허용가능한 담체 또는 희석제로는 물, 생리 식염수, 크림, 로션, 각종 형태의 겔 및 단쇄 알코올 및 글리콜(예: 에틸 알코올 및 프로필렌 글리콜) 등을 포함할 수 있으나, 이에 한정되는 것은 아니다.In addition, the composition of the present invention may be formulated in the form of an external preparation for skin suitable for direct administration to a treatment site. In this case, a physiologically acceptable carrier or diluent suitable for topical administration to the skin may be further included. Such physiologically acceptable carriers or diluents may include, but are not limited to, water, physiological saline, creams, lotions, gels of various types and short-chain alcohols and glycols such as ethyl alcohol and propylene glycol .
또한, 본 발명은 다른 측면으로, 태반 추출물을 유효성분으로 포함하는 근육 기능 개선용 식품 조성물을 제공한다.In another aspect, the present invention provides a food composition for improving muscle function comprising placenta extract as an active ingredient.
본 발명에 따른 식품 조성물은 기능성 식품(functional food), 영양 보조제(nutritional supplement), 건강식품(health food), 식품 첨가제(food additives), 사료, 및 이의 조합으로 이루어진 군에서 선택되는 어느 하나의 제형을 가질 수 있다.The food composition according to the present invention may be any one of formulations selected from the group consisting of functional food, nutritional supplement, health food, food additives, feed, and combinations thereof. Lt; / RTI >
이하, 본 발명을 하기 실시예에 의하여 더욱 상세하게 설명한다. 단, 하기 실시예는 본 발명을 예시하기 위한 것일 뿐, 본 발명의 범위가 이들만으로 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
실시예Example 1. 태반 추출물의 제조 1. Preparation of Placenta Extract
하기 실험에서 사용된 태반 추출물은 녹십자웰빙에서 제공되는 라이넥(Laennec)의 주사 제형을 사용하였으며, 상기 라이넥은 하기의 방법으로 제조하였다. 먼저, 자하거(인간 태반)를 아세톤 처리하여 탈지한 후, 불완전 가수분해물이 생성되지 않도록 펩신 및 염산 처리로 충분히 가수분해하여 태반 추출물인 라이넥을 제조하였다. 상기 라이넥을 적용하기 위하여, 주사 제형의 라이넥을 v/v%로 기본 배지에 희석하여 사용하였다. 이때, 배지는 DMEM (Dulbeco's Modified Eagle's Media) 배지를 사용하였고, 1% 페니실린을 첨가하여 사용하였다.The placenta extract used in the following experiment was the injection form of Laennec provided in Green Cross Wellness, and the lyne was prepared by the following method. First, the human placenta was degreased with acetone treatment, and hydrolyzed sufficiently by treatment with pepsin and hydrochloric acid to prevent incomplete hydrolyzate production, thereby preparing a placenta extract, linecn. To apply the lynech, the linac of the injectable formulation was diluted in v / v% to the basal medium. At this time, the medium used was DMEM (Dulbeco's Modified Eagle's Media) medium and 1% penicillin was added thereto.
실험예Experimental Example 1. 산화 스트레스 상황에서 근육 세포에 대한 태반 추출물의 효과 확인 1. Determination of the effect of placental extract on muscle cells in oxidative stress
세포 생존율 측정 방법 중 하나인 Cell Counting Kit-8(CCK-8) 분석 기법 및 형광 프로브 H2DCFDA를 사용하여 산화 스트레스 상황에서 근육 세포에 대한 태반 추출물의 효과를 확인하였다. Using the Cell Counting Kit-8 (CCK-8) assay and the fluorescent probe H2DCFDA, one of the methods for measuring cell viability, the effect of placental extract on muscle cells in oxidative stress was examined.
단일 세포로 현탁 되어있는 C2C12 세포를 96-웰 플레이트에 웰당 1x104개의 세포를 시딩(seeding)한 후, 37℃에서 24시간동안 CO2 배양기 안에서 배양하였다. 배양 후, 각각 다음과 같이 처리하였다: i) H2O2 500 uM를 최고 농도로 설정하여 1/2 희석으로 최종 128배 희석한 농도까지 C2C12 배지에 24시간동안 처리하였다; ii) 라이넥 10%를 최고 농도로 설정하여 1/2 희석으로 최종 128배 희석한 농도까지 C2C12 배지에 24시간동안 처리하였다; iii) C2C12 배지에 라이넥 5%를 24시간동안 처리한 후, H2O2를 4시간동안 처리하였다. 그 다음, CCK-8 용액을 각각의 농도로 H2O2 처리된 C2C12 배지와 1:10 비율로 희석한 뒤 37℃에서 1시간동안 CO2 배양기 안에서 배양하였다.C2C12 cells suspended in a single cell were seeded on a 96-well plate at 1 × 10 4 cells per well and cultured in a CO 2 incubator at 37 ° C. for 24 hours. After incubation, each was treated as follows: i) treated with C2C12 medium for 24 hours to a final dilution of 128 by 1/2 dilution with 500 uM H 2 O 2 set at the highest concentration; ii) treated with C2C12 medium for 24 hours to a final dilution of 128-fold with 1/2 dilution with 10% linac set to the highest concentration; iii) The C2C12 medium was treated with 5% linoleum for 24 hours and then treated with H 2 O 2 for 4 hours. The CCK-8 solution was then diluted 1: 10 with C2C12 medium treated with H 2 O 2 at each concentration and incubated at 37 ° C for 1 hour in a CO 2 incubator.
이후, 분광 광도계(spectrophotometer)를 이용하여 450 nm 파장에서 흡광도를 측정하였다. 이때, reference는 450 nm로 하였다. 각 군의 세포 생존율은 대조군(Con)과 비교하여 백분율(%)로 계산하였다. 그 결과를 도 1a 내지 도 1c에 나타내었다.Then, the absorbance was measured at a wavelength of 450 nm using a spectrophotometer. At this time, the reference was set to 450 nm. Cell viability of each group was calculated as percentage (%) compared with control (Con). The results are shown in Figs. 1A to 1C.
상기 단계 iii)에서 C2C12 배지에 라이넥 5% 및 H2O2를 처리한 후, ROS(reactive oxygen species) 프로브인 20,70-dichlorodihydroflurescein(H2DCFDA)를 30분동안 처리하여 세포 내의 세포질을 ROS 염색하였다. 이후, 유세포 분석기를 통해 세포 내 과산화수소량을 측정하여 마이크로플레이트 리더기(Molecular Devices, USA)를 이용하여 495 nm 및 525 nm 파장에서 형광량을 측정함으로써 세포질 내 H2O2 질량을 측정하였다. 그 결과를 도 1d에 나타내었다.In step iii), the C2C12 medium was treated with 5% linoleic acid and H 2 O 2 , and then the ROS (reactive oxygen species) probe, 20,70-dichlorodihydroflurescein (H2DCFDA) Respectively. Then, by using a flow cytometer cell measures the amount of hydrogen peroxide in a microplate reader to measure the fluorescence amount at 495 nm and 525 nm wavelengths using a (Molecular Devices, USA) cytoplasm in H 2 O 2 Mass was measured. The results are shown in Fig. 1d.
산화 스트레스에 의한 C2C12 세포의 사멸을 측정한 결과, 농도가 증가할수록 세포 생존율이 감소하는 것을 확인하였고, 이는 산화 스트레스가 세포에 대해 세포 독성을 보이는 것을 의미한다. 또한, 각각의 해당 농도를 24시간동안 처리한 후 세포 생존율의 절반에 해당하는 농도는 250 uM임을 알 수 있었다. 상기 농도를 이후 실험에 적용하였다 (도 1a). 인간 태반 추출물이 C2C12 세포 독성 및 생존율에 미치는 영향을 확인하기 위하여 실시한 실험에서, 라이넥의 농도가 증가할수록 세포 생존율이 증가하는 것을 확인하였고, 이는 라이넥이 농도 10%까지는 세포에 대한 독성이 없고, 세포 성장 효과를 가지는 것을 의미한다 (도 1b).As a result of measuring the death of C2C12 cells by oxidative stress, it was confirmed that the cell survival rate decreases with increasing concentration, which means that oxidative stress is cytotoxic to the cells. Furthermore, it was found that the concentration corresponding to half of the cell survival rate after 250 hours treatment of each of the respective concentrations was 250 uM. This concentration was then applied to the experiment (FIG. 1A). In order to confirm the effect of human placenta extract on C2C12 cell toxicity and survival rate, it was confirmed that the cell viability was increased with increasing concentration of ligne, which was not toxic to cells up to 10% , And has a cell growth effect (Fig. 1B).
또한, 산화 스트레스에 의한 C2C12 세포 사멸에 대하여 인간 태반 추출물의 보호 작용을 확인한 결과, 라이넥을 전처리한 집단에서 H2O2 단독 처리군에 비해 비교군 대비 유의하게 생존율이 증가하는 것을 확인하였다. 이는 인간 태반 추출물이 산화 스트레스로부터 세포를 보호하는 것을 의미한다 (도 1c). 또한, 인간 태반 추출물의 C2C12 세포의 세포질 내의 ROS 제거 능력을 확인하기 위해 실시한 실험에서, 라이넥을 전처리한 스트레스 유도 집단의 세포질 내 ROS가 스트레스 단독 유도 집단에 비하여 유의하게 감소하는 것을 확인하였다. 이는 인간 태반 추출물이 세포 내 ROS를 감소하는 기능을 가지고 있음을 의미한다 (도 1d).In addition, the protective effect of human placenta extract against C2C12 cell death by oxidative stress was confirmed to be significantly higher in the group pretreated with linac than in the group treated with H 2 O 2 alone. This means that the human placenta extract protects cells from oxidative stress (Fig. 1C). In addition, in an experiment conducted to confirm ROS removal ability in the cytoplasm of C2C12 cells of the human placenta extract, it was confirmed that the intracellular ROS of the stress inducing group pretreated with the lynek was significantly reduced compared to the stress alone group. This means that the human placenta extract has a function of reducing intracellular ROS (FIG. 1d).
실험예Experimental Example 2. 세포 내 미토콘드리아 합성과 태반 추출물의 상관관계 2. Relationship between intracellular mitochondria synthesis and placenta extract
웨스턴블롯 분석 기법(western blot analysis)을 사용하여 세포 내 미토콘드리아 합성에 관여하는 단백질의 발현량을 측정함으로써 산화 스트레스 상황에서 태반 추출물의 역할을 확인하였다.Western blot analysis was used to determine the role of placenta extract in oxidative stress by measuring the amount of protein expression involved in intracellular mitochondrial synthesis.
C2C12 세포를 96-웰 플레이트에 웰당 1x104 세포를 시딩한 후, 라이넥 5%를 24시간동안 처리하였고 산화적 스트레스를 유도하기 위해 H2O2를 4시간동안 처리하였다. 이후, 시약이 포함된 모든 배지를 워시 아웃 한 후, 세포 스크래퍼를 이용하여 배양된 세포를 수집하였다. 수집된 세포를 포스파타제 억제제가 포함된 PRO-PREP 단백질 추출 용액(Intron Biotechnology, South korea)와 혼합하였다. 상기 혼합 용액을 얼음 위에서 30분간 인큐베이션한 후, 12,000 rpm으로 20분동안 원심 분리하였다. 이후, 단백질이 포함된 상층액을 수득한 후, BCA 단백질 분석 키트(Thermo Fisher Scientific, USA)를 이용하여 단백질을 정량하였다.C2C12 cells were seeded in 96-well plates at 1x10 4 cells per well, treated with 5% linoleum for 24 hours and treated with H 2 O 2 for 4 hours to induce oxidative stress. Thereafter, all of the medium containing the reagent was washed out, and the cultured cells were collected using a cell scraper. The collected cells were mixed with a PRO-PREP protein extract solution containing a phosphatase inhibitor (Intron Biotechnology, South Korea). The mixed solution was incubated on ice for 30 minutes and then centrifuged at 12,000 rpm for 20 minutes. Thereafter, the supernatant containing the protein was obtained, and the protein was quantified using a BCA protein assay kit (Thermo Fisher Scientific, USA).
30 ㎍의 단백질들을 100℃에서 5분동안 가열하였고, 환원제가 포함된 12% 아크릴아마이드 겔을 이용하여 SDS-PAGE 분석(sodium dodecyl sulfate poly acrylamide gel electrophoresis assay)을 통해 가열된 각 단백질들을 전기영동 하였다. 전기영동을 통해 아크릴아마이드겔 상에서 크기 별로 분리된 단백질들을 웨스턴블롯 기법을 사용하여 PVDF(poly-vinyl difluoride) membrane으로 이동시켰다. 이후, membrane이 다른 단백질로 오염되는 것을 막기 위해, 5% 탈지유(skim milk)를 사용하여 블로킹(blocking)하였다. 화학발광 이미지 시스템(chemiluminescence imaging system)을 이용하여 membrane을 관찰하였고, Image J 1.38 버전 소프트웨어를 이용하여 데이터를 분석하였다. 그 결과를 도 2a에 나타내었다.30 의 of proteins were heated at 100 캜 for 5 minutes, and each protein heated by SDS-PAGE analysis (sodium dodecyl sulfate polyacrylamide gel electrophoresis assay) was electrophoresed using a 12% acrylamide gel containing a reducing agent . Proteins separated in size on acrylamide gels by electrophoresis were transferred to a poly-vinyl difluoride (PVDF) membrane using Western blotting. Thereafter, the membrane was blocked with 5% skim milk to prevent contamination with other proteins. Membranes were observed using a chemiluminescence imaging system and data were analyzed using Image J 1.38 version software. The results are shown in Fig.
또한, 미토콘드리아의 질량을 측정하기 위해, mito-tracker를 사용하여 상기 라이넥 5% 또는 H2O2를 처리한 C2C12 세포의 미토콘드리아를 형광 염색하였다. 이후, 마이크로 플레이트를 사용하여 형광량을 측정하였고, 대조군 대비 상대 값으로 표기하였다. 그 결과를 도 2b에 나타내었다.Further, the mitochondria of C2C12 cells with a mito-tracker processes the license or
도 2a에 나타난 바와 같이, 미토콘드리아 신생합성의 핵심인자로 밝혀진 SIRT1, P-AMPK, PGC1-alpha 단백질의 발현량이 라이넥을 처리한 두 집단에서 처리하지 않은 두 집단에 비해 증가함을 확인하였다. 이는 인간 태반 추출물이 미토콘드리아 신생합성의 마스터 단백질인 PGC1-alpha의 발현량을 AMPK에 의존하여 증가시킨다는 것을 의미한다. 또한, 도 2b에 나타난 바와 같이, mito-tracker를 이용하여 각 집단의 미토콘드리아 질량을 비교한 결과, 라이넥을 처리한 두 집단에서 처리하지 않은 두 집단에 비해 증가함을 확인하였다. 이는 마스터 단백질인 PGC1-alpha의 발현량이 미토콘드리아 질량과 직접적인 상관관계가 있음을 의미한다.As shown in FIG. 2A, the expression levels of SIRT1, P-AMPK and PGC1-alpha proteins, which were found to be key factors for mitochondrial neogenesis, were found to be increased in the two groups treated with linacs compared to the two groups not treated with linacs. This means that the human placenta extract increases the expression level of PGC1-alpha, the master protein of mitochondrial neogenesis, depending on AMPK. In addition, as shown in FIG. 2B, the mitochondrial masses of the respective groups were compared using the mito-tracker, and it was confirmed that the two groups treated with the linecn were increased in comparison with those of the untreated group. This indicates that the expression level of the master protein, PGC1-alpha, has a direct correlation with the mitochondrial mass.
실험예Experimental Example 3. 세포의 3. Cellular 오토파지Auto Phage (( autophagyautophagy ) 조절에 태반 추출물이 미치는 영향Effect of Placenta Extract on Control
웨스턴블롯 분석 기법을 사용하여 오토파지 마커 단백질의 발현량을 측정함으로써 태반 추출물이 세포의 오토파지 조절에 미치는 영향을 확인하였다.Western blot analysis was used to determine the effect of placenta extract on the autophagy of the cells by measuring the expression level of the autophage marker protein.
상기 실시예 2의 실험과 동일한 방법으로 C2C12 세포에 라이넥 5% 및 H2O2를 처리한 후, 세포를 용해시켜 세포의 용해물을 수득하였고, 인간 태반 추출물이 오토파지 조절에 미치는 영향을 확인하기 위해 웨스턴블롯 분석 기법을 사용하여 오토파지 마커인 LC3 및 Beclin-1 단백질의 발현량을 비교하였다. 또한, 노인성 근 감소증 환자의 경우 오토파지가 적절치 않게 조절되어 근육 세포 내의 p62 단백질이 축적된다는 이전 연구에 근거하여, 인간 태반 추출물이 스트레스 상황에서 플럭스 조절에 미치는 영향을 확인하기 위해 p62 단백질의 발현량을 평가하였다. 그 결과를 도 3에 나타내었다.C2C12 cells were treated with 5% linoleic acid and H 2 O 2 in the same manner as in Example 2, and cells were lysed to obtain cell lysates. The effect of human placenta extract on autophagy control We used Western blot analysis to compare the expression levels of the autophage markers LC3 and Beclin-1 protein. In order to examine the effect of human placenta extract on the flux regulation in the stress state, the expression level of p62 protein was determined based on the previous study that autophagy was inappropriately regulated and accumulation of p62 protein in muscle cells in the senile dyslipidemia patients . The results are shown in Fig.
도 3에 나타난 바와 같이, 라이넥 5%를 전처리한 집단에서 처리하지 않은 집단에 비해 오토파지 마커인 LC3 및 Beclin-1 두 단백질 모두 발현량이 증가한 것을 확인하였다. 또한, 스트레스 환경(H2O2 처리 집단)에서 LC3 II 밴드가 I 밴드에 비해 발현량이 증가한 것을 확인하였다. 이는 인간 태반 추출물이 오토파지를 상향조절 할 뿐만 아니라, 스트레스 상황에서 플럭스를 원활하게 조절한다는 것을 의미한다. 결과적으로, 인간 태반 추출물이 스트레스 상황에서 오토파지 플럭스를 적절하게 조절하여 p62 단백질의 축적을 감소시킨다는 것을 확인할 수 있었다.As shown in FIG. 3, the expression levels of both the LC3 and Beclin-1 proteins, which are autophagy markers, were increased in the group treated with 5% linac, compared with the group not treated with the 5% linac. In the stress environment (H 2 O 2 Treated group), the expression level of LC3 II band was increased compared with that of I band. This means that the human placenta extract not only up-regulates the auto-phage but also smoothly controls the flux under stress conditions. As a result, it was confirmed that the human placenta extract properly regulates the autophage flux under stress conditions, thereby reducing accumulation of p62 protein.
실험예Experimental Example 4. 4. 근지배력에On my power 대한 태반 추출물의 효과 확인 Confirmation of the effect of placenta extract
근육 볼륨에 미치는 영향이 신경근 차단의 회복에 의하여 일어날 수 있는 변수를 확인하기 위하여 신경근 지배 시험법을 사용함으로써 관찰 초기와 마지막 날짜에 근전도 변화를 확인하였다.Effects on muscle volume To confirm the variables that could be caused by the restoration of the nerve root block, we used the neuromuscular control test method to confirm the EMG changes at the initial and last days of observation.
생후 6주된 무모생쥐(중앙실험동물, 서울, 한국) 18마리를 구입하여 Normal, BTX 처리군 및 BTX와 라이넥 처리군으로 나누어 실험을 진행하였다. 이때, 18마리의 무모생쥐는 각 군에 대해 무작위로 선택하여 배정하였다. 총 관찰기간 중 7일 후, 30 μl의 식염수로 희석한 보툴리눔 톡신(Allergan, USA) 0.5 단위를 인슐린 실린지를 이용하여 BTX 처리군 및 BTX와 라이넥 처리군 무모생쥐의 우측 뒷다리 비복근에 주입하였다. 본 실험에서 음식과 물은 자율식이로 진행하였으며, 독성 유발 7일 전부터 16일 후까지 3.6 ml/kg의 태반 추출물을 주 3회 우측 뒷다리 비복근에 피하주사 하였다. 이후, EMG(electromyography) 모니터링 장치를 이용하여 관찰 5일차 및 16일차에 근전도를 측정하였다. 이때, EMG 측정은 다리 근육을 조절하는 척추 신경을 전기적으로 자극한 후, 활성 신호를 받은 전극으로부터 전극의 반응 크기를 측정하여 수행하였다. 그 결과를 도 4a 및 도 4b에 나타내었다.Sixteen male rehabilitated mice (Central laboratory animals, Seoul, Korea) were purchased from 18 rats and divided into normal, BTX, BTX, and laneect. At this time, 18 hairless mice were randomly selected for each group. Seven days after the total observation period, 0.5 units of botulinum toxin (Allergan, USA) diluted in 30 μl of saline was injected into the BTX treated group and the right hindlimb gastrocnemius muscle of BTX and lynx treated group reared mice using an insulin syringe. In this experiment, food and water proceeded autonomously. The placenta extract of 3.6 ml / kg was injected subcutaneously in the right hind gland gastrocnemius muscle three times a week from 7 days before toxicity to 16 days after toxicity. Then, EMG (electromyography) monitoring device was used to measure EMG on the 5th day and 16th day of observation. At this time, the EMG measurement was performed by electrically stimulating the vertebral nerve regulating the leg muscles, and measuring the reaction magnitude of the electrode from the electrode receiving the activation signal. The results are shown in Figs. 4A and 4B.
도 4a 및 도 4b에 나타난 바와 같이, 보툴리눔 톡신을 주입한 BTX 처리군 및 BTX와 라이넥 처리군 모두 대조군(Normal)에 비해 떨어진 근 지배력을 확인하였고, 두 군간의 차이는 발견되지 않았다. 이를 통해, 근 볼륨이 차이가 나는 것은 신경 재지배로 이루어지는 것이 아닌, 태반 추출물에 의하여 발생하는 것임을 확인할 수 있었다.As shown in FIGS. 4A and 4B, BTX treatment group injected with botulinum toxin, and BTX and linac treated group were observed to have less muscle power than the control group, and no difference was found between the two groups. These results suggest that the difference in muscle volume is caused by placenta extract rather than nerve regeneration.
실험예Experimental Example 5. 근 위축 모델에서 태반 추출물의 효과 5. Effect of Placenta Extract on the Atrophy Model
실체현미경과 PRIMOSLITE 5.8E 버전 소프트웨어를 사용하여 무모생쥐의 비복근을 육안적으로 평가하고 3차원적인 부피를 비교하여 측정함으로써 근 위축에 대한 태반 추출물의 보호효과를 확인하였다.Using a stereomicroscope and PRIMOS LITE 5.8E version software, we evaluated the protective effects of the placenta extract against muscle atrophy by grossly evaluating the gastrocnemius muscle of uninjured mice and comparing the three dimensional volume.
상기 실험예 4에서 근전도 변화를 관찰한 후, 관찰 마지막 날에 각 집단의 무모생쥐를 희생하여 우측 뒷다리에서 피부를 떼어내고 실체현미경을 이용하여 비복근을 육안적으로 평가함으로써 근육 부피를 비교하였다. 그 결과를 도 5a에 나타내었다. 또한, 각 집단의 무모생쥐 우측 뒷다리의 비복근을 잘라 PRIMOSLITE 소프트웨어를 이용하여 스캔함으로써 비복근의 질량 및 볼륨을 측정하였다. 이때, 볼륨 측정은 자동적으로 '0'으로 설정되도록 하여 평가를 진행하였으며, 생쥐 한 마리당 3번씩 측정하여 평균을 결과값으로 나타내었다. 그 결과를 도 5b 및 도 5c에 나타내었다.After observing EMG changes in Experimental Example 4, the skin was removed from the right hind leg at the sacrifice of hairless mice in each group on the last day of observation, and muscle volume was compared by grossly evaluating the gastrocnemius using a stereomicroscope. The results are shown in Fig. 5A. We also measured the mass and volume of the gastrocnemius muscle by scanning the gastrocnemius muscle of the right hind limb of each group using PRIMOS LITE software. At this time, the evaluation was performed by setting the volume measurement to '0' automatically, and the average value was measured by measuring three times per mouse. The results are shown in Figs. 5B and 5C.
도 5a 내지 도 5c에 나타난 바와 같이, 라이넥을 처리한 집단에서 처리하지 않은 집단에 비해 근 볼륨이 크게 유지됨을 확인하였다.As shown in FIGS. 5A to 5C, it was confirmed that the muscle volume was maintained to a large extent in the group treated with the linac, compared with the group not treated.
Claims (8)
상기 약학적 조성물의 제형이 주사제, 크림, 젤, 패치, 분무제, 연고제, 경고제, 로션제, 마이크로니들, 롤러, 및 이의 조합으로 이루어진 군으로부터 선택되는 어느 하나인 것을 특징으로 하는, 약학적 조성물.The method according to claim 1,
Wherein the formulation of the pharmaceutical composition is any one selected from the group consisting of injections, creams, gels, patches, sprays, ointments, alerts, lotions, microneedles, rollers, and combinations thereof. .
상기 식품 조성물이 기능성 식품, 영양 보조제, 건강식품, 식품 첨가제, 사료, 및 이의 조합으로 이루어진 군으로부터 선택되는 어느 하나의 제형을 갖는 것을 특징으로 하는, 식품 조성물.8. The method of claim 7,
Wherein the food composition has any one of the formulations selected from the group consisting of a functional food, a nutritional supplement, a health food, a food additive, a feed, and a combination thereof.
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| WO2021182913A1 (en) * | 2020-03-12 | 2021-09-16 | (주)녹십자웰빙 | Composition, comprising placenta extract, for preventing or treating liver disease and improving liver function |
| KR20220161415A (en) | 2020-04-14 | 2022-12-06 | (주)녹십자웰빙 | Antiviral composition containing placenta extract |
| KR20220164524A (en) | 2020-04-14 | 2022-12-13 | (주)녹십자웰빙 | Antiviral composition containing microRNA derived from placental extract |
| KR20220167294A (en) | 2020-04-14 | 2022-12-20 | (주)녹십자웰빙 | Antiviral composition containing placenta-derived material |
| WO2023085456A1 (en) * | 2021-11-10 | 2023-05-19 | (주)녹십자웰빙 | Placenta hydrolysate and method for producing same |
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| KR100642080B1 (en) * | 2006-02-11 | 2006-11-13 | 케이에이취팜 주식회사 | A food composition containing swine placenta extraction for improving menopausal disorder, health foods comprising of it and preparation method thereof |
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| WO2021182913A1 (en) * | 2020-03-12 | 2021-09-16 | (주)녹십자웰빙 | Composition, comprising placenta extract, for preventing or treating liver disease and improving liver function |
| KR20220161415A (en) | 2020-04-14 | 2022-12-06 | (주)녹십자웰빙 | Antiviral composition containing placenta extract |
| KR20220164524A (en) | 2020-04-14 | 2022-12-13 | (주)녹십자웰빙 | Antiviral composition containing microRNA derived from placental extract |
| KR20220167294A (en) | 2020-04-14 | 2022-12-20 | (주)녹십자웰빙 | Antiviral composition containing placenta-derived material |
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