WO2021068802A1 - Application of nadh and salt and/or nmn thereof in preparation of drug or health care product for enhancing muscle or inhibiting muscle volume reduction - Google Patents
Application of nadh and salt and/or nmn thereof in preparation of drug or health care product for enhancing muscle or inhibiting muscle volume reduction Download PDFInfo
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- 150000003839 salts Chemical class 0.000 title claims abstract description 47
- 210000003205 muscle Anatomy 0.000 title claims abstract description 38
- 239000003814 drug Substances 0.000 title claims abstract description 14
- 229940079593 drug Drugs 0.000 title claims abstract description 13
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 10
- 238000002360 preparation method Methods 0.000 title claims abstract description 6
- 238000011038 discontinuous diafiltration by volume reduction Methods 0.000 title abstract 3
- 230000002708 enhancing effect Effects 0.000 title abstract 3
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims abstract description 47
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 claims abstract description 47
- 239000000203 mixture Substances 0.000 claims abstract description 16
- 210000001665 muscle stem cell Anatomy 0.000 claims description 13
- 238000005728 strengthening Methods 0.000 claims description 7
- 230000007423 decrease Effects 0.000 claims description 6
- BAWFJGJZGIEFAR-NNYOXOHSSA-O NAD(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-O 0.000 claims description 5
- 238000007911 parenteral administration Methods 0.000 claims description 5
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 230000001028 anti-proliverative effect Effects 0.000 claims description 2
- 230000017531 blood circulation Effects 0.000 claims description 2
- 230000002438 mitochondrial effect Effects 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 102000004169 proteins and genes Human genes 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 claims description 2
- 230000003716 rejuvenation Effects 0.000 claims description 2
- 108020005087 unfolded proteins Proteins 0.000 claims description 2
- 201000010099 disease Diseases 0.000 abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 239000004480 active ingredient Substances 0.000 abstract 1
- 238000001647 drug administration Methods 0.000 abstract 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- WOVKYSAHUYNSMH-RRKCRQDMSA-N 5-bromodeoxyuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(Br)=C1 WOVKYSAHUYNSMH-RRKCRQDMSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- WOVKYSAHUYNSMH-UHFFFAOYSA-N BROMODEOXYURIDINE Natural products C1C(O)C(CO)OC1N1C(=O)NC(=O)C(Br)=C1 WOVKYSAHUYNSMH-UHFFFAOYSA-N 0.000 description 5
- 229950004398 broxuridine Drugs 0.000 description 5
- 230000020411 cell activation Effects 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- SXEHKFHPFVVDIR-UHFFFAOYSA-N [4-(4-hydrazinylphenyl)phenyl]hydrazine Chemical compound C1=CC(NN)=CC=C1C1=CC=C(NN)C=C1 SXEHKFHPFVVDIR-UHFFFAOYSA-N 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 241000700159 Rattus Species 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 230000008836 DNA modification Effects 0.000 description 2
- 102000016359 Fibronectins Human genes 0.000 description 2
- 108010067306 Fibronectins Proteins 0.000 description 2
- -1 NMN Chemical compound 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 108010039918 Polylysine Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- FZAQROFXYZPAKI-UHFFFAOYSA-N anthracene-2-sulfonyl chloride Chemical compound C1=CC=CC2=CC3=CC(S(=O)(=O)Cl)=CC=C3C=C21 FZAQROFXYZPAKI-UHFFFAOYSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 210000002808 connective tissue Anatomy 0.000 description 2
- 238000001085 differential centrifugation Methods 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- 210000001087 myotubule Anatomy 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 229920000656 polylysine Polymers 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 210000000689 upper leg Anatomy 0.000 description 2
- 206010028289 Muscle atrophy Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000011888 autopsy Methods 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 230000020763 muscle atrophy Effects 0.000 description 1
- 201000000585 muscular atrophy Diseases 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 230000000276 sedentary effect Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 239000000021 stimulant Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7084—Compounds having two nucleosides or nucleotides, e.g. nicotinamide-adenine dinucleotide, flavine-adenine dinucleotide
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
Provided is an application of NADH and a salt and/or NMN thereof in the preparation of a drug or health care product for enhancing muscle or inhibiting muscle volume reduction. Provided is a drug composition or health care product composition for enhancing muscle or inhibiting muscle volume reduction, containing NADH and a salt and/or NMN thereof, and containing no active ingredients other than NADH and salts and/or NMN thereof. The NADH and salts and/or NMN thereof may be administered enterally or parenterally, and the routes of administration are broad and convenient. The invention is suitable for persons having insufficient exercise and middle-aged and elderly persons, and according to the severity of the disease condition, the drug administration route and dosage are adjusted.
Description
本申请涉及药物或保健品技术领域，尤其涉及NADH及其盐和/或NMN在制备肌肉增强或抑制肌肉量下降的药物或保健品中的应用。This application relates to the technical field of medicines or health care products, in particular to the application of NADH and its salts and/or NMN in the preparation of medicines or health care products for muscle strengthening or inhibiting muscle loss.
在长期运动不足以及年长的人群中，会发生肌肉量的减少，对于老年人和长期久坐不动的人来说，进行足以增加肌肉量的运动是极其困难的。因此这类人群存在期望增加肌肉量或抑制肌肉量下降的情况。另外，运动员等相关人群也存在增强肌肉的需求，除了运动，还会借助于有效的营养保健品或药品，以求更快更高效地达到目的。In long-term under-exercise and elderly people, muscle mass loss will occur. For the elderly and sedentary people, it is extremely difficult to exercise sufficient to increase muscle mass. Therefore, there are situations in such people who desire to increase muscle mass or inhibit muscle mass decline. In addition, athletes and other related groups also have a need to strengthen muscles. In addition to exercise, they will also rely on effective nutrition and health products or drugs to achieve their goals faster and more efficiently.
目前，常用的肌肉增强剂一般为甾类和生长激素等，但这类药物属于兴奋剂或激素类药物，对身体具有副作用，不能长期服用。At present, commonly used muscle-enhancing agents are generally steroids and growth hormones, but these drugs are stimulants or hormone drugs, which have side effects on the body and cannot be taken for a long time.
为了解决上述问题，本申请的技术目的之一在于提供NADH（Nicotinamide adenine dinucleotide，还原型烟酰胺腺嘌呤二核苷酸）及其盐和/或NMN（β-Nicotinamide
Mononucleotide，β-烟酰胺单核苷酸）在制备增强肌肉或抑制肌肉量下降的药物或保健品中的应用。In order to solve the above problems, one of the technical objectives of this application is to provide NADH (Nicotinamide adenine dinucleotide, reduced nicotinamide adenine dinucleotide) and its salt and/or NMN (β-Nicotinamide
Mononucleotide, β-nicotinamide mononucleotide) in the preparation of muscle-strengthening or inhibiting the application of muscle loss drugs or health products.
在本申请的其中一个实施例中，当同时使用NADH及其盐和NMN时，两者的摩尔比例为(1-10):(1-10)，优选为(1-3):(1-3)。In one of the embodiments of the present application, when NADH and its salt and NMN are used at the same time, the molar ratio of the two is (1-10): (1-10), preferably (1-3): (1- 3).
在本申请的其中一个实施例中，NADH及其盐和/或NMN为肠内给药或肠外给药均可，其中肠内给药主要为口服给药，相应地，所述肠内给药的剂型为片剂、粉剂、胶囊剂、颗粒剂、丸剂、混悬剂、糖浆剂、合剂、散剂和滴丸中的任意一种或几种；肠外给药主要为注射给药、肺吸收或局部给药，相应地，所述肠外给药的剂型为注射剂、软膏剂、栓剂和吸入剂中的任意一种或几种。 In one of the embodiments of the present application, NADH and its salts and/or NMN can be administered either enterally or parenterally, wherein the enteral administration is mainly oral administration. Accordingly, the enteral administration The dosage form of the medicine is any one or more of tablets, powders, capsules, granules, pills, suspensions, syrups, mixtures, powders and dripping pills; parenteral administration is mainly injection, pulmonary For absorption or local administration, correspondingly, the dosage form for parenteral administration is any one or more of injections, ointments, suppositories and inhalants. To
在本申请的其中一个实施例中，肠内给药尤其是口服给药时NADH及其盐的单次剂量为0.1~10mg/kg，优选为0.1~4mg/kg。日次数为1~3次，日用量为0.1~30mg/kg，优选是0.1~12mg/kg，更优选0.4~2mg/kg。肠外给药尤其是静脉注射时NADH及其盐的单次剂量为0.1~2mg/kg，优选是0.1~1mg/kg。其中kg为对象个体的体重单位。In one of the embodiments of the present application, the single dose of NADH and its salt during enteral administration, especially oral administration, is 0.1-10 mg/kg, preferably 0.1-4 mg/kg. The daily frequency is 1 to 3 times, and the daily dosage is 0.1 to 30 mg/kg, preferably 0.1 to 12 mg/kg, more preferably 0.4 to 2 mg/kg. The single dose of NADH and its salt during parenteral administration, especially intravenous injection, is 0.1-2 mg/kg, preferably 0.1-1 mg/kg. Wherein kg is the weight unit of the subject.
在本申请的其中一个实施例中，肠内给药尤其是口服给药时NMN的日用量为2~30mg/kg，优选是2~10mg/kg；肠外给药尤其是静脉注射时日剂量为5~10mg/Kg。其中kg为对象个体的体重单位。In one of the embodiments of the present application, the daily dosage of NMN for enteral administration, especially oral administration, is 2-30 mg/kg, preferably 2-10 mg/kg; parenteral administration, especially the daily dosage for intravenous injection It is 5~10mg/Kg. Wherein kg is the weight unit of the subject.
在本申请的其中一个实施例中，所述NADH及其盐和/或NMN的应用范围包括但不限于增肌、塑肌、肌肉保持、治疗医学上诊断为肌肉萎缩及与其相关的病症。In one of the embodiments of the present application, the scope of application of NADH and its salts and/or NMN includes but is not limited to muscle gain, muscle building, muscle maintenance, treatment of medically diagnosed muscle atrophy and related diseases.
本申请的技术方案中，利用NADH及其盐和/或NMN增加肌肉量或抑制肌肉量下降，是基于NADH及其盐或NMN在生物体内可以生成NAD+，而NAD+可以促进肌肉内的毛细血管密度增加，增加血流量，并且还可以引起线粒体非折叠蛋白和抗增殖蛋白的表达，从而使肌肉干细胞恢复活力。In the technical solution of the present application, the use of NADH and its salts and/or NMN to increase muscle mass or inhibit muscle mass decline is based on the fact that NADH and its salts or NMN can generate NAD+ in the organism, and NAD+ can promote the capillary density in the muscle Increase, increase blood flow, and can also cause the expression of mitochondrial unfolded proteins and antiproliferative proteins, thereby rejuvenating muscle stem cells.
本申请的技术目的之二在于提供一种增强肌肉或抑制肌肉量下降的药物组合物或保健品组合物，所述组合物中以NADH及其盐和/或NMN为单一活性组分。The second technical purpose of the present application is to provide a pharmaceutical composition or a health care product composition for strengthening muscles or inhibiting muscle loss, in which NADH and its salts and/or NMN are the single active components.
在本申请的其中一个实施例中，上述组合物中同时包含NADH及其盐和NMN时，两者的摩尔比例为(1-10):(1-10)，优选为(1-3):(1-3)。In one of the embodiments of the present application, when NADH and its salt and NMN are included in the above composition, the molar ratio of the two is (1-10): (1-10), preferably (1-3): (1-3).
在本申请的其中一个实施例中，上述组合物还包括药学上或食品中可接受的辅料。In one of the embodiments of the present application, the above-mentioned composition further includes pharmaceutically or food acceptable excipients.
实施本申请实施例，将具有如下有益效果：Implementing the embodiments of this application will have the following beneficial effects:
本申请公开了NADH及其盐和/或NMN在制备肌肉增强或抑制肌肉量下降的药物或保健品中的应用，服用含NADH及其盐和/或NMN的保健品或药物可有效地增强肌肉或防止肌肉量降低，适用于运动不足人群及中老年人群，希望保持肌肉量的情况，可依据病情的严重程度，调整施药途径和施药量。可以长期服用，对人体无副作用。This application discloses the application of NADH and its salts and/or NMN in the preparation of drugs or health products for muscle strengthening or inhibiting muscle loss. Taking health products or drugs containing NADH and its salts and/or NMN can effectively strengthen muscles Or to prevent the decrease of muscle mass, it is suitable for people with insufficient exercise and middle-aged and elderly people. If you want to maintain muscle mass, you can adjust the route and dosage of the drug according to the severity of the disease. It can be taken for a long time without side effects on the human body.
下面将结合具体实施例对本申请的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本申请一部分实施例，而不是全部的实施例。基于本申请中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本申请保护的范围。The technical solutions of the present application will be clearly and completely described below in conjunction with specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in this application, all other embodiments obtained by a person of ordinary skill in the art without creative work shall fall within the protection scope of this application.
实施例1Example 1
NADH及其盐和/或NMN的体外实验效果：In vitro experimental effects of NADH and its salts and/or NMN:
从6月龄的雄性SD大鼠摘出大腿部肌肉组织，将脂肪及结缔组织等去除或切碎后，在37℃下用1.25mg/mL蛋白酶水溶液处理1小时，通过差速离心分离将肌纤维碎片等去除，播种到涂布有聚赖氨酸及纤连蛋白的板上，在5％CO
2气氛中、37℃下进行24小时培养，之后用PBS进行清洗，由此分离出肌肉干细胞。培养基使用添加有10％马血清的DMEM(达尔伯克改良伊格尔培养基，Dulbecco’s Modified Eagle Medium)，将该培养基简记为“10％HS-DMEM”。
The thigh muscle tissue was extracted from a 6-month-old male SD rat, the fat and connective tissue were removed or minced, and then treated with 1.25 mg/mL protease aqueous solution at 37°C for 1 hour, and the muscle fibers were separated by differential centrifugation The debris and the like are removed, seeded on a plate coated with polylysine and fibronectin, cultured in a 5% CO 2 atmosphere at 37° C. for 24 hours, and then washed with PBS to isolate muscle stem cells. As the medium, DMEM (Dulbecco's Modified Eagle Medium) supplemented with 10% horse serum was used, and the medium was abbreviated as "10% HS-DMEM".
分别将NADH及其盐、NMN、NADH及其盐:NMN（1:1）的组合物溶解于水中后，添加至10％HS-DMEM，制备包含NADH及其盐和/或NMN的10％HS-DMEM，NADH及其盐和/或NMN的浓度为2.5μg/mL。将分离肌肉干细胞的培养基更换为包含NADH及其盐和/或NMN的10％HS-DMEM，进行24小时培养，并使用水(即不含NADH及其盐和NMN的10%HS-DMEM)作为空白对照。Dissolve NADH and its salt, NMN, NADH and its salt: NMN (1:1) composition in water, and add it to 10% HS-DMEM to prepare 10% HS containing NADH and its salt and/or NMN -The concentration of DMEM, NADH and its salts and/or NMN is 2.5 μg/mL. Replace the culture medium for separating muscle stem cells with 10% HS-DMEM containing NADH and its salts and/or NMN, culture for 24 hours, and use water (ie 10% HS-DMEM without NADH and its salts and NMN) As a blank control.
在自培养结束2小时前，向培养基中添加溴脱氧尿苷(BrdU)以使终浓度成为10μM。培养结束后，使用经水冷的添加有0.1％H
2O
2的甲醇将经NADH及其盐和/或NMN处理的肌肉干细胞在4℃下固定10分钟。进一步使用2N盐酸在37℃下进行1小时DNA改性处理，再使用抗BrdU抗体作为一次抗体，使用HRP结合抗小鼠IgG抗体作为二次抗体，用二氨基联苯胺(DAB)使其显色，对BrdU阳性细胞进行检测。求出BrdU阳性细胞在总细胞数中所占的比例，为肌肉干细胞活化率。将结果示于表1。
Two hours before the end of the culture, bromodeoxyuridine (BrdU) was added to the medium so that the final concentration became 10 μM. After the incubation, the muscle stem cells treated with NADH and its salts and/or NMN were fixed at 4°C for 10 minutes using water-cooled methanol added with 0.1% H 2 O 2. Further use 2N hydrochloric acid at 37°C for 1 hour of DNA modification treatment, then use anti-BrdU antibody as the primary antibody, use HRP-conjugated anti-mouse IgG antibody as the secondary antibody, and use diaminobenzidine (DAB) to make it color , To detect BrdU positive cells. The ratio of BrdU-positive cells in the total number of cells is calculated, and it is the activation rate of muscle stem cells. The results are shown in Table 1.
如表1结果所示，与空白对照相比，NADH及其盐、NMN、NADH及其盐:NMN（1:1）组合物的干细胞活化率均高于空白对照组，并且在浓度相同的条件下，效果上NADH及其盐＞NADH及其盐:NMN（1:1）组合物＞NMN，这说明NADH及其盐在人体中NAD+的转化率要高于NMN。As shown in the results of Table 1, compared with the blank control, the stem cell activation rate of NADH and its salt, NMN, NADH and its salt: NMN (1:1) composition are all higher than that of the blank control group, and under the same concentration conditions Next, the effect is NADH and its salt>NADH and its salt: NMN (1:1) composition>NMN, which shows that the NAD+ conversion rate of NADH and its salt in the human body is higher than that of NMN.
实施例2Example 2
NADH及其盐和/或NMN的体内实验效果：In vivo experimental effects of NADH and its salts and/or NMN:
将14周龄的雄性SD大鼠以体重为基准、以每组6只分为对照组、NADH及其盐给药组、NMN给药组和NADH及其盐/NMN组合物给药组4组。对各组每只鼠按体重以500mg/kg的标准分别1天给药1次水、NADH及其盐、NMN和NADH及其盐/NMN组合物，进行7天口服给药。此外，在解剖2小时前以50mg/kg体重的比例腹腔内给药溴脱氧尿苷。Fourteen-week-old male SD rats were divided into four groups: control group, NADH and its salt administration group, NMN administration group, and NADH and its salt/NMN composition administration group based on body weight and 6 per group. . Each mouse in each group was given water, NADH and its salt, NMN and NADH and its salt/NMN composition once a day at the standard of 500 mg/kg body weight, and was orally administered for 7 days. In addition, bromodeoxyuridine was intraperitoneally administered at a rate of 50 mg/kg body weight 2 hours before the autopsy.
从上述大鼠摘出大腿部肌肉组织，将脂肪及结缔组织等去除或切碎后，在37℃下用1.25mg/mL蛋白酶水溶液处理1小时。通过差速离心分离将肌纤维碎片等去除后，播种到涂布有聚赖氨酸及纤连蛋白的板上，在5％CO
2气氛中37℃下进行24小时培养，用PBS进行清洗，分离出肌肉干细胞。作为培养基，使用上述实施例1中使用的10％HS-DMEM。
The thigh muscle tissue was extracted from the above-mentioned rat, fat and connective tissue, etc. were removed or minced, and then treated with a 1.25 mg/mL protease aqueous solution at 37°C for 1 hour. After removing muscle fiber fragments by differential centrifugation, they are seeded on a plate coated with polylysine and fibronectin, incubated in a 5% CO 2 atmosphere at 37°C for 24 hours, washed with PBS, and separated Out muscle stem cells. As the medium, the 10% HS-DMEM used in Example 1 above was used.
使用经水冷的添加有0.1％H
2O
2的甲醇将经分离的肌肉干细胞在4℃下固定10分钟。进一步使用2N盐酸在37℃下进行1小时DNA改性处理后，使用抗BrdU抗体作为一次抗体、使用HRP结合抗小鼠IgG抗体作为二次抗体，用二氨基联苯胺(DAB)使其显色，对BrdU阳性细胞进行检测。求出BrdU阳性细胞在总细胞数中所占的比例，作为肌肉干细胞活化率。此外，以对照组的肌肉干细胞活化率为基准，将各组的肌肉干细胞活化率的比作为肌肉干细胞活化率。将结果示于表2。
The isolated muscle stem cells were fixed at 4°C for 10 minutes using water-cooled methanol added with 0.1% H 2 O 2. After further DNA modification treatment with 2N hydrochloric acid at 37°C for 1 hour, anti-BrdU antibody was used as the primary antibody, HRP-conjugated anti-mouse IgG antibody was used as the secondary antibody, and the color was developed with diaminobenzidine (DAB) , To detect BrdU positive cells. The ratio of BrdU-positive cells to the total number of cells was calculated as the activation rate of muscle stem cells. In addition, based on the muscle stem cell activation rate of the control group, the ratio of the muscle stem cell activation rate of each group was taken as the muscle stem cell activation rate. The results are shown in Table 2.
如表2所示的结果，与对照组相比，其他三组的肌肉干细胞活化效果分别为2.2~2.5倍，说明其相对于对照组具有显著的活化效果，说明NADH及其盐和/或NMN在肌肉增强或抑制肌肉量下降方面具有显著的作用。As shown in Table 2, compared with the control group, the muscle stem cell activation effects of the other three groups were 2.2~2.5 times, respectively, indicating that it has a significant activation effect compared to the control group, indicating that NADH and its salts and/or NMN It has a significant role in muscle strengthening or inhibiting the decrease of muscle mass.
以上所揭露的仅为本申请较佳实施例而已，当然不能以此来限定本申请之权利范围，因此依本申请权利要求所作的等同变化，仍属本申请所涵盖的范围。The above-disclosed are only preferred embodiments of this application, and of course the scope of rights of this application cannot be limited by this. Therefore, equivalent changes made in accordance with the claims of this application still fall within the scope of this application.
Claims (10)
- NADH及其盐和/或NMN在制备增强肌肉或抑制肌肉量下降的药物或保健品中的应用。The application of NADH and its salt and/or NMN in the preparation of medicines or health care products for strengthening muscles or inhibiting the decrease of muscle mass.
- 根据权利要求1所述的应用，其特征在于，所述NADH及其盐和/或NMN增加肌肉量或抑制肌肉量下降，是基于NADH及其盐或NMN在生物体内可以生成NAD+，NAD+可以促进肌肉内的毛细血管密度增加，增加血流量，还引起线粒体非折叠蛋白和抗增殖蛋白的表达，使肌肉干细胞恢复活力。The application according to claim 1, wherein the NADH and its salts and/or NMN increase muscle mass or inhibit the decrease of muscle mass based on the fact that NADH and its salts or NMN can generate NAD+ in the organism, and NAD+ can promote The density of capillaries in the muscle increases, increasing blood flow, and also causing the expression of mitochondrial unfolded proteins and antiproliferative proteins, rejuvenating muscle stem cells.
- 根据权利要求1所述的应用，其特征在于，当同时使用NADH及其盐和NMN时，两者的摩尔比例为（1-10）:（1-10）。The application according to claim 1, characterized in that when NADH and its salt and NMN are used at the same time, the molar ratio of the two is (1-10): (1-10).
- 根据权利要求3所述的应用，其特征在于，当同时使用NADH及其盐和NMN时，两者的摩尔比例为（1-3）:（1-3）。The application according to claim 3, characterized in that when NADH and its salt and NMN are used at the same time, the molar ratio of the two is (1-3): (1-3).
- 根据权利要求1所述的应用，其特征在于，所述NADH及其盐和/或NMN为肠内给药或肠外给药。The application according to claim 1, wherein the NADH and its salts and/or NMN are administered enteral or parenterally.
- 根据权利要求5所述的应用，其特征在于，肠内给药时NADH及其盐的单次剂量为0.1~10mg/kg，肠外给药时NADH及其盐的单次剂量为0.1~2mg/kg，每日给药1~3次。The application according to claim 5, wherein the single dose of NADH and its salt is 0.1-10 mg/kg when administered enterally, and the single dose of NADH and its salt is 0.1-2 mg when administered parenterally. /kg, administer 1 to 3 times a day.
- 根据权利要求5所述的应用，其特征在于，肠内给药时NMN的日剂量为2~30mg/kg，肠外给药时，日剂量为5~10mg/Kg。The application according to claim 5, wherein the daily dose of NMN is 2-30 mg/kg for enteral administration, and 5-10 mg/Kg for parenteral administration.
- 一种增强肌肉或抑制肌肉量下降的药物组合物或保健品组合物，所述组合物除NADH及其盐和/或NMN以外不含其它活性组分。A pharmaceutical composition or a health care product composition for strengthening muscles or inhibiting muscle loss, said composition containing no other active components except NADH and its salts and/or NMN.
- 根据权利要求8所述的药物组合物或保健品组合物，其特征在于，组合物中同时包含NADH及其盐和NMN时，两者的摩尔比例为（1-10）:（1-10）。The pharmaceutical composition or health care product composition according to claim 8, wherein when the composition contains both NADH and its salt and NMN, the molar ratio of the two is (1-10): (1-10) .
- 根据权利要求8所述的药物组合物或保健品组合物，其特征在于，其还包括药学上或食品中可接受的辅料。The pharmaceutical composition or health care product composition according to claim 8, characterized in that it further comprises pharmaceutically or food acceptable excipients.
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