CN104367587A - Application of nicotinamide mononucleotide in preparation of medicines for promotion of nerve regeneration after cerebral ischemia - Google Patents
Application of nicotinamide mononucleotide in preparation of medicines for promotion of nerve regeneration after cerebral ischemia Download PDFInfo
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- CN104367587A CN104367587A CN201310655963.0A CN201310655963A CN104367587A CN 104367587 A CN104367587 A CN 104367587A CN 201310655963 A CN201310655963 A CN 201310655963A CN 104367587 A CN104367587 A CN 104367587A
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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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Abstract
The invention relates to the technical field of medicines. The present invention provides application of nicotinamide mononucleotide in preparation of medicines for promotion of nerve regeneration after cerebral ischemia, and the nicotinamide mononucleotide can be used as an active pharmaceutical ingredient and a pharmaceutically acceptable supplementary material for preparation of medicine compositions. The nicotinamide mononucleotide compound itself is an endogenous protective substance, and the available data do not report the adverse reaction, so that the nicotinamide mononucleotide as a medicine or a health product is high in safety and wide in therapeutic window.
Description
Technical field
The present invention relates to medical art, be specifically related to the application of nicotinamide mononucleotide. in preparation promotion neuranagenesis medicine or health food.
Background technology
Cerebral infarction is a kind of commonly encountered diseases of serious harm human health, and its disability rate is very high.Apoplexy survivor often leaves over dysfunction in various degree, has a strong impact on the quality of life of patient, brings heavy burden to society and family.Research finds, carries out scientific and rational intervention can effectively reduce case fatality rate and disability rate to cerebral stroke at acute period correlative factor.Traditional view is thought, after cerebral ischemia occurs,
The neuron of the necrosis because of ischemia can not restore or regenerate because nerve exist only in period of embryo and birth is early stage.But recent researches finds to there is a large amount of neural stem cell at the olfactory bulb of adults, Hippocampus and tricorn, and these stem cell can be divided into neurocyte in life process, and this is found to be nervous system injury and disease brings new treatment concept.Neural stem cell can be divided into the various neurocyte such as neuron, neurogliocyte under incentive condition, and this process is subject to the regulation and control of endogenous and extrinsic factor.
According to the record of state food and drug administration, in the market not for the medicine promoting neuranagenesis after cerebral ischemia.
Nicotinamide mononucleotide. (Nicotinamide mononucleotide is called for short NMN) is the endogenous material existed in a kind of body, can carry out iii vitro chemical synthesis at present.Current research finds that it has and falls hypoglycemic effect (see document: Yoshino J; et al.; Nicotinamide mononucleotide; a keyNAD (+) intermediate; treats the pathophysiology of diet-and age-induceddiabetes in mice, Cell Metab.2011Oct5; 14 (4): 528-36.).Whether nicotinamide mononucleotide. can be promoted Differentiation of Neural Stem Cells and be moved, improve function of nervous system, improve survival, quality of life aspect there is not yet bibliographical information.
Summary of the invention
The object of the present invention is to provide the novelty teabag of nicotinamide mononucleotide., namely provide nicotinamide mononucleotide. to promote the application in neuranagenesis medicine or health food in preparation.。
The present inventor, through lot of experiments, surprisingly finds: nicotinamide mononucleotide. has and promotes neuranagenesis effect, and effectively can improve the functional of the cranial nerve caused because of ischemia.The present invention adopts intraluminal middle cerebral artery occlusion in rats blocking-up to prepare cerebral ischemic model, has inquired into the impact of this compound of nicotinamide mononucleotide. on the neuranagenesis in 7-14 after ischemia days further.
The invention provides the application of nicotinamide mononucleotide. in the medicine or health food of preparation promotion neuranagenesis.
Invention further provides nicotinamide mononucleotide. promotes in the medicine of neuranagenesis after cerebral ischemia or health food application in preparation.
Nicotinamide mononucleotide. of the present invention is the nicotinamide mononucleotide. of this area routine, and it is the endogenous material existed in human body, also can synthetic, or is the commodity of direct commercial gained.The chemical structural formula of described nicotinamide mononucleotide. is as shown in formula I:
Nicotinamide mononucleotide. of the present invention can be prepared into pharmaceutical composition as active pharmaceutical ingredient and pharmaceutically acceptable adjuvant.Wherein, described active pharmaceutical ingredient nicotinamide mononucleotide. is the aforementioned nicotinamide mononucleotide. of the present invention.Wherein said pharmaceutically acceptable adjuvant refers to the excipient substance of art of pharmacy routine.Described pharmaceutically acceptable adjuvant comprises: diluent, excipient, binding agent, filler and burst apart in agent one or more, described excipient substance also comprises flavor substance.Wherein said excipient is preferably water; Described binding agent is preferably: cellulose derivative, gelatin or polyvinylpyrrolidone; Described filler is preferably starch, and described disintegrating agent is preferably: calcium carbonate or sodium bicarbonate.Described flavor substance refers to taste for improving pharmaceutical composition or local flavor and the material added, and described flavor substance preferably comprises various flavouring agent, essence and/or sweeting agent.
The dosage form of wherein said pharmaceutical composition is this area regular dosage form.The dosage form of described pharmaceutical composition, according to the composition of its route of administration or excipient substance, preferably comprises: tablet, powder, capsule, injection, drop or transdermal gel preparation.
The route of administration of wherein said pharmaceutical composition preferably comprises: oral administration, drug administration by injection, mucosal drug delivery or transdermal administration.The mode of wherein said drug administration by injection preferably comprises: injection in lumbar injection, subcutaneous injection, intravenous injection, intramuscular injection or lymph node.
In described pharmaceutical composition, the content of nicotinamide mononucleotide. is preferably 0.1% ~ 99.9%, and its content is preferably 0.5 ~ 99.9%, and described percentage ratio is mass percent.
The individuality of described medicine composite for curing is preferably human or animal, and the using dosage of described pharmaceutical composition is preferably 1 ~ 1000mg/ kg body weight/sky, and concrete use amount suitably can adjust according to the age of Different Individual, the state of an illness etc.
The present invention is except special instruction, and described percentage ratio is all mass percent.
On the basis meeting this area general knowledge, above-mentioned each optimum condition, can combination in any, obtains the preferred embodiments of the invention.Agents useful for same of the present invention and raw material are all commercially.
Positive progressive effect of the present invention is:
1, the invention provides nicotinamide mononucleotide. (NMN) promotes in the medicine of neuranagenesis after cerebral ischemia or health food application in preparation.Described compound effectively can promote that more polyneural stem cell is to ripe neural cellular differentiation, improve nervous function damage caused by brain diseases, make up the neurocyte lost in above-mentioned lysis, significantly promote learning and memory function, recover to be significant to neurological impairments.
2, nicotinamide mononucleotide. route of administration kind comparatively horn of plenty, can be applied to by oral, injection or the approach such as mucosal drug delivery the individuality needing treatment according to different dosage form.
3, nicotinamide mononucleotide. compound itself is body Endogenous protective substances, and available data is not yet reported for work its untoward reaction, therefore it can be used as medicine or health product safety high, treatment window width.
Accompanying drawing explanation
Fig. 1 is the hippocampal dentate district BrdU/DCX Positive Cell Counts figure giving nicotinamide mononucleotide. group mice after cerebral ischemic model group and cerebral ischemia;
Wherein A is Double immune fluorescent dyeing schematic diagram; B is cartogram; BrdU/DCX positive cell is newborn migration neuron (arrow logo).
Fig. 2 is the hippocampal dentate district BrdU/NeuN Positive Cell Counts figure giving nicotinamide mononucleotide. group mice after cerebral ischemic model group and cerebral ischemia;
Wherein A is Double immune fluorescent dyeing schematic diagram; B is cartogram; BrdU/NeuN positive cell is newborn mature neuron (arrow logo).
Fig. 3 is the 14 days learning and memory function comparison diagrams giving nicotinamide mononucleotide. group mice after cerebral ischemic model group and cerebral ischemia;
Wherein A is water maze laboratory schematic diagram; B is for escaping the experiment statistics figure that hides; C is the long-distance cartogram in path.
Fig. 4 is the proliferative conditions of In vitro culture neural stem cell after giving control drug and nicotinamide mononucleotide.;
Wherein A is immunofluorescence dyeing schematic diagram; B is cartogram.
Detailed description of the invention
Now in conjunction with the embodiments and accompanying drawing, the present invention is described in detail, but enforcement of the present invention is not limited only to this.
Agents useful for same of the present invention and raw material all commercially maybe can be prepared by literature method.The experimental technique of unreceipted actual conditions in the following example, usually conveniently condition, or according to the condition that manufacturer advises.
Embodiment 1
Arteries and veins in mouse brain is adopted to block preparation brain damage model, model preparation process is as follows: by mice (C57BL/6J, be purchased from Shanghai Si Laike laboratory animal company limited) anesthesia (4% chloral hydrate, 0.1ml/10g, lumbar injection) (1. mice selects 5-0 monofilament nylon suture, and the final diameter on top is 0.150 ~ 0.200mm to implement apoplexy operation afterwards.Veutro cervical region median incision, further blunt separation sternohyoid, breastbone Papillary muscle and omohyoid, expose right carotid (CCA) and right side external carotid artery (ECA).2. the near-end of CCA and the section start of external carotid artery 3-0 suture ligature.3. internal carotid artery (ICA) to relax ligation with 3-0 suture, and determines that it does not have branch to be tied.4. 3mm place under common carotid artery bifurcated, with the ligation that 6-0 suture is lax, in order to labelling arteriotomy position and stop hemorrhage.5. arteriotomy place is near at hand the position of 6-0 suture.6. bolt line carefully injects to intracranial, and the degree of depth of insertion is counted from crotch, mice 9-11mm.Now, slight resistance shows that the top of bolt line has been positioned at the near-end of anterior cerebral artery, blocks the origin of ipsilateral cerebral medium-sized artery and posterior communicating artery.) apoplexy takes out and fasten line Reperfu-sion for 2 hours.
Administration group lumbar injection nicotinamide mononucleotide. (NMN) (being purchased from Sigma Co., USA) 500mg/kg/day (normal saline) successive administration 9 days; Continuous 9 days of matched group injection normal saline.All animals were at 3rd ~ 5 days lumbar injection 5-bromodeoxyuridine nucleoside (5-bromodeoxyuridine, Brdu is purchased from Sigma company) 300mg/kg, got brain and cooks frozen section and carry out immunohistochemical staining after paraformaldehyde heart perfusion is fixing in the 9th day.
Newborn neuron proliferation apoptosis adopts 5-bromodeoxyuridine nucleoside antibody (BrdU is purchased from Abcam company) and two cortin antibody (doublecortin is purchased from Santa Cruz company) to carry out Double immunofluorescence detection; Newborn mature neuron adopts 5-bromodeoxyuridine nucleoside antibody (BrdU is purchased from Abcam company) and neuronal glutamate transporter antibody (neuron NeuN is purchased from Millipore company) to carry out Double immunofluorescence detection.The detection method of Double immunofluorescence can refer to: Pei Wang; Chao-YuMiao; et al.Induction of autophagy contributes to the neuroprotection ofnicotinamide phosphoribosyltransferase in cerebral ischemic stroke; Autophagy8:1,1 – 11; January1,2012.
As shown in Figure 1, nicotinamide mononucleotide. (NMN) effectively can increase newborn migration neuron number.As shown in Figure 2, nicotinamide mononucleotide. (NMN) can significantly improve the ripe neuronal quantity of new life in mouse brain Post stroke Hippocampus DG district.Neuranagenesis after these results explanation nicotinamide mononucleotide. can promote ischemia on animal brain ischemia model.
Embodiment 2
Adopt arteries and veins in mouse brain to block preparation brain damage model, model preparation process is with embodiment 1.
Administration group lumbar injection nicotinamide mononucleotide. (NMN) (being purchased from Sigma Co., USA) 500mg/kg/day (normal saline) successive administration 9 days; Continuous 9 days of matched group injection normal saline.
After 14 days, the learning and memory function of Morris water maze laboratory to two groups of mices is used to evaluate.Morris water maze laboratory step is as follows: mice is faced pool wall and puts into water by (1), puts into East, West, South, North four direction at random as original position.Record mice finds the time of underwater platform (second) and the path (rice) finding platform.In front training several times, if mice finds the time of underwater platform more than 60 seconds, then need guiding mice to find platform and be positioned on platform to stop 10 seconds.(2) mice is fetched, dry after be placed on electric filament lamp under dry, put back in cage.Every mice trains 4 times every day, and twice training time is spaced apart 15 ~ 20 minutes, continuously training 5 days.Record is escaped latent time and is escaped path and carry out statistical analysis to it.
As shown in Figure 3, nicotinamide mononucleotide. (NMN) effectively can strengthen the learning and memory function after focal cerebral ischemia, illustrates that this medicine has the effect promoting nerve regeneratl.
Embodiment 3
Adopt the neural stem cell of In vitro culture, matched group gives normal saline, and nicotinamide mononucleotide. (NMN is purchased from Sigma company) administration, dosage is 300 μMs.
The neural stem cell process of In vitro culture is as follows: get pregnant Mus (pregnant age 14-15 days) cervical dislocation and put to death, and after cleaning, takes out tire Mus and is also separated cerebral tissue, under anatomic microscope, peel meninges and blood vessel off, and cut brain.With microsurgical scissors, shred cerebral tissue, and drip 2ml Accutase digestive enzyme, put into cell culture incubator, digestion 15-20 minute.After digestion, add equal-volume nerve stem cell culture medium and stop digestion.After 200 order nylon wires filter, abandon supernatant, collecting cell after centrifugal 10 minutes for 1000 revs/min.After cell is resuspended, be inoculated in nerve stem cell culture medium.Put into 37 DEG C, in 5% CO2 gas incubator, cultivate 5-7 days.The cell clone of the non-adherent growth of collected by centrifugation, fully piping and druming makes it form individual cells.The neural ball of monoclonal within Secondary Culture 4-7 days, can be obtained.
As shown in Figure 4, nicotinamide mononucleotide. (NMN) effectively can strengthen the propagation of mouse neural stem cells, illustrates that this medicine has the effect promoting nerve regeneratl.
Below the preferred embodiment of the invention is illustrated, but the invention is not limited to described embodiment, those of ordinary skill in the art also can make all equivalent modification or replacement under the prerequisite without prejudice to the invention spirit, and these equivalent modification or replacement are all included in the application's claim limited range.
Claims (2)
1. the application of nicotinamide mononucleotide. in the medicine or health food of preparation promotion neuranagenesis.
2. nicotinamide mononucleotide. promotes the application in the medicine of neuranagenesis after cerebral ischemia or health food in preparation.
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105030807A (en) * | 2015-07-13 | 2015-11-11 | 中国人民解放军第二军医大学 | Application of nicotinamide riboside to preparing medicine for treating cerebral ischemic diseases |
| WO2017185549A1 (en) * | 2016-07-30 | 2017-11-02 | 邦泰生物工程(深圳)有限公司 | Method for preparing nicotinamide mononucleotide 2 |
| CN107412247A (en) * | 2016-05-23 | 2017-12-01 | 上海风劲生物医药科技有限公司 | Application of the nicotinamide mononucleotide in prevention or treatment cerebral hemorrhage medicine is prepared |
| US10392415B2 (en) | 2015-10-02 | 2019-08-27 | Metro International Biotech, Llc | Crystal forms of β-nicotinamide mononucleotide |
| CN110446780A (en) * | 2017-01-31 | 2019-11-12 | 东方酵母工业株式会社 | Proliferation of pluripotent stem cells promotor |
| US10548913B2 (en) | 2015-08-05 | 2020-02-04 | Metro International Biotech, Llc | Nicotinamide mononucleotide derivatives and their uses |
| US10618927B1 (en) | 2019-03-22 | 2020-04-14 | Metro International Biotech, Llc | Compositions and methods for modulation of nicotinamide adenine dinucleotide |
| CN111759877A (en) * | 2020-07-02 | 2020-10-13 | 深圳市旷逸生物科技有限公司 | NMN-containing pharmaceutical composition for improving intelligence and strengthening brain |
| CN112105718A (en) * | 2018-03-22 | 2020-12-18 | 东方酵母工业株式会社 | Pluripotent stem cell differentiation promoter |
| US11180521B2 (en) | 2018-01-30 | 2021-11-23 | Metro International Biotech, Llc | Nicotinamide riboside analogs, pharmaceutical compositions, and uses thereof |
| US11939348B2 (en) | 2019-03-22 | 2024-03-26 | Metro International Biotech, Llc | Compositions comprising a phosphorus derivative of nicotinamide riboside and methods for modulation of nicotinamide adenine dinucleotide |
| US11952396B1 (en) | 2021-05-27 | 2024-04-09 | Metro International Biotech, Llc | Crystalline solids of nicotinic acid mononucleotide and esters thereof and methods of making and use |
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| CN101601679A (en) * | 2009-03-17 | 2009-12-16 | 中国人民解放军第二军医大学 | A kind of application of nicotinamide mononucleotide. |
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2013
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Patent Citations (1)
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| CN101601679A (en) * | 2009-03-17 | 2009-12-16 | 中国人民解放军第二军医大学 | A kind of application of nicotinamide mononucleotide. |
Cited By (21)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105030807B (en) * | 2015-07-13 | 2018-11-06 | 中国人民解放军第二军医大学 | Niacinamide ribose is preparing the application in treating Imaging in Patients with Cerebral Ischemia Disease drug |
| CN105030807A (en) * | 2015-07-13 | 2015-11-11 | 中国人民解放军第二军医大学 | Application of nicotinamide riboside to preparing medicine for treating cerebral ischemic diseases |
| US11464796B2 (en) | 2015-08-05 | 2022-10-11 | Metro International Biotech, Llc | Nicotinamide mononucleotide derivatives and their uses |
| US10548913B2 (en) | 2015-08-05 | 2020-02-04 | Metro International Biotech, Llc | Nicotinamide mononucleotide derivatives and their uses |
| US10392415B2 (en) | 2015-10-02 | 2019-08-27 | Metro International Biotech, Llc | Crystal forms of β-nicotinamide mononucleotide |
| US10392416B2 (en) | 2015-10-02 | 2019-08-27 | Metro International Biotech, Llc | Crystal forms of beta-nicotinamide mononucleotide |
| US11059847B2 (en) | 2015-10-02 | 2021-07-13 | Metro International Biotech, Llc | Crystal forms of β-nicotinamide mononucleotide |
| CN107412247A (en) * | 2016-05-23 | 2017-12-01 | 上海风劲生物医药科技有限公司 | Application of the nicotinamide mononucleotide in prevention or treatment cerebral hemorrhage medicine is prepared |
| WO2017185549A1 (en) * | 2016-07-30 | 2017-11-02 | 邦泰生物工程(深圳)有限公司 | Method for preparing nicotinamide mononucleotide 2 |
| CN108026130A (en) * | 2016-07-30 | 2018-05-11 | 邦泰生物工程(深圳)有限公司 | A kind of method 2 for preparing nicotinamide mononucleotide |
| US11040996B2 (en) | 2016-07-30 | 2021-06-22 | Bontac Bio-Engineering(Shenzhen) Co., Ltd | Method for preparing nicotinamide mononucleotide (NMN) |
| CN110446780A (en) * | 2017-01-31 | 2019-11-12 | 东方酵母工业株式会社 | Proliferation of pluripotent stem cells promotor |
| CN110446780B (en) * | 2017-01-31 | 2024-01-09 | 东方酵母工业株式会社 | Multipotent stem cell proliferation promoter |
| US11634690B2 (en) | 2017-01-31 | 2023-04-25 | Oriental Yeast Co., Ltd. | Agent for accelerating growth of pluripotent stem cells |
| US11180521B2 (en) | 2018-01-30 | 2021-11-23 | Metro International Biotech, Llc | Nicotinamide riboside analogs, pharmaceutical compositions, and uses thereof |
| US11814652B2 (en) | 2018-03-22 | 2023-11-14 | Oriental Yeast Co., Ltd. | Pluripotent stem cell differentiation-promoting agent |
| CN112105718A (en) * | 2018-03-22 | 2020-12-18 | 东方酵母工业株式会社 | Pluripotent stem cell differentiation promoter |
| US10618927B1 (en) | 2019-03-22 | 2020-04-14 | Metro International Biotech, Llc | Compositions and methods for modulation of nicotinamide adenine dinucleotide |
| US11939348B2 (en) | 2019-03-22 | 2024-03-26 | Metro International Biotech, Llc | Compositions comprising a phosphorus derivative of nicotinamide riboside and methods for modulation of nicotinamide adenine dinucleotide |
| CN111759877A (en) * | 2020-07-02 | 2020-10-13 | 深圳市旷逸生物科技有限公司 | NMN-containing pharmaceutical composition for improving intelligence and strengthening brain |
| US11952396B1 (en) | 2021-05-27 | 2024-04-09 | Metro International Biotech, Llc | Crystalline solids of nicotinic acid mononucleotide and esters thereof and methods of making and use |
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