CN108125988A - Ginkolide B is in the application that microglia inflammatory reaction is inhibited to mitigate Alzheimer disease symptoms - Google Patents
Ginkolide B is in the application that microglia inflammatory reaction is inhibited to mitigate Alzheimer disease symptoms Download PDFInfo
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- CN108125988A CN108125988A CN201711500596.1A CN201711500596A CN108125988A CN 108125988 A CN108125988 A CN 108125988A CN 201711500596 A CN201711500596 A CN 201711500596A CN 108125988 A CN108125988 A CN 108125988A
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- bifidobacterium breve
- ginkolide
- alzheimer disease
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
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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
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/14—Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
Abstract
The invention discloses the composition of the treatment Alzheimer disease of bilobalide-containing B and the composition and preparation method thereof and application methods.The active constituent of the composition is ginkolide B, further includes bifidobacterium breve active factors and stabilizer.The invention demonstrates that ginkolide B can improve the cognitive function of Alzheimer disease and inhibit Neuroinflammation, and the composition materials safety is nontoxic, preparation method is simple, cheap, easy to utilize.
Description
Technical field
The present invention relates to the purposes of ginkolide B, and in particular to the preparation of bilobalide-containing composition B and its treatment Ah
Application in the drug of Alzheimer's disease, belongs to field of biological pharmacy.
Background technology
Alzheimer disease (Alzheimer ' sdisease, AD) is a kind of irreversible nervus retrogression disease of progressive
Disease, major pathologic features show as outer amyloid protein (A β) deposition of nerve cell and are formed in senile plaque (SP) and nerve cell
Protein tau Hyperphosphorylationof forms neurofibrillary tangles (NFT).Excessively generating and accumulating for A β is considered as AD main pathogenic
Factor can result in injury of mitochondria, cause oxidative stress, destroy nerve synapse function, induce inflammatory reaction etc..Wherein,
More and more concerns are caused by the A β Neuroinflammations induced, different pathological stages of the cascade of response of inflammation in AD all show
The important feature gone out.Current research thinks that AD is a kind of inflammation damnification of central nervous system, the A β depositions of brain exception and god
It is triggering agent through first fibre matting, activates patient's intracerebral cascade of response of inflammation.On the one hand, the amyloid fiber that intracerebral is gradually assembled
The malnutrition tangled and neuron can be caused by the inflammatory mediator of Deiter's cells secretion.The neuroglia being activated for a long time
Cell is by discharging such as reactive oxygen intermediates, nitric oxide, proteolytic enzyme, complementary factor or excitatory amino acid toxicity
Substance further damages peripheral nerve member.In addition, the various inflammatory mediators of brain secretion can increase A under the action of other conditions
The production quantity of β promotes A beta-aggregations.Microglia is main effector cell in neuroinflamation generating process.In AD patient and
Transgenic animals intracerebral, using A β as the microglia that assembles the neuron of denaturation around the senile plaque of core and largely activate.A
β can directly activate microglia, and proinflammatory enzyme (iNOS and COX-2) and proinflammatory cytokine can be generated after overactivity
(TNF-α, IL-1 β and IL-6).These inflammatory mediators can damage the neuron of surrounding, and it is dead to eventually lead to its denaturation.It is in addition, dead
The neuron died can also further make Activated Microglia, generate more inflammatory factors, these inflammatory mediators can induce more
More Activated Microglias leads to local neure damage, and can promote the generation of A β, constitutes pernicious loop.To sum up
Described, immune reacted with inflammation damnification being persistently activated of intracerebral plays very important work in the occurrence and development of AD
With.In the research process of anti-inflammatory drug, non-steroidal anti-inflammatory drugs shows to delay and treats the potentiality of AD, but due to side effect
Interference, affect its Clinical practice.Ginkgolides is special in ginkgo biloba p.e (Ginkgo biloba extract, EGb)
Some complicated compounds of one kind, animal experiments show that ginkgolides can be by reducing nerve cell apoptosis, protection nerve
Member.Ginkgolides is generally acknowledged platelet-activating factor antagonist, wherein the activity of ginkolide B (ginkgolide B, GB)
It is most strong, there is anti-inflammatory, neuroprotection, maintain a variety of effects such as blood brain barrier integrity, have to neurodegenerative diseases such as AD
Significance.Chinese patent CN 104042607B disclose pharmaceutical composition for treating dementia and preparation method thereof, institute
It states pharmaceutical composition and includes any one of Bilobalide and ginkalide A, C or two kinds of combination, but it does not use ginkgo
Lactone B.Research finds that GB inhibits the SH-SY5Y cell neurotoxicities effect of A β and PrPC induction.In addition, GB can pass through
PI3K/Akt approach adjusts the metabolism of the intracellular A amyloid beta-protein precursors (APP) of SH-SY5Y, and it is horizontal to reduce APP in endochylema.Though
Though right ginkolide B to neuroprotective and its prevent cranial vascular disease research it has been reported that but precise mechanism still not
Very understand.In recent years, by clinical observation and practice repeatedly, it is believed that AD is under the jurisdiction of the traditional Chinese medical science " network disease " scope, the traditional Chinese medical science
Interpretation of the cause, onset and process of an illness key may be " poison damage brain network ".GB is during AD is treated except the dependent interaction link that early period is verified is in addition to target spot, going back
There may be the inflammatory reactions for reducing A β depositions, alleviating intracerebral, protect brain neuron cell from damage, block intracerebral inflammation
The vicious circle of damage.It, will if its inner link with the traditional Chinese medical science " poison damage brain network " can be found for above-mentioned inflammation damnification process
To distinguish that controlling AD provides direct basis from " poison damage brain network " angle.It yet there are no and be used for ginkolide B and bifidobacterium breve
Treat correlative study and the report of Alzheimer disease.
Invention content
The defects of it is an object of the invention to overcome the prior art and deficiency, inventive application ginkolide B and short bifid
The application of the disease of bacillus active factors composition treatment Alzheimer disease.The object of the present invention is to provide it is a kind of it is safe,
The composition of effective treatment Alzheimer disease.
A kind of composition for being used to treat Alzheimer disease, the composition include ginkolide B, bifidobacterium breve activity
The factor and glyceryl monostearate, each component weight ratio are：
Ginkolide B：85-100 parts by weight；
Bifidobacterium breve active factors：5-10 parts by weight；
Glyceryl monostearate：5-15 parts by weight；
The preparation method of the bifidobacterium breve active factors is as follows：
(1) prepared by seed liquor：Picking single bacterium colony bifidobacterium breve (Bifidobacterium breve) LJM-006, preservation
Number is CGMCC No.5418, is inoculated in sterilized MRS fluid nutrient mediums, 37 DEG C, and Anaerobic culturel is for 24 hours to get to short pair
Discrimination bacillus LJM-006 seed liquors；
(2) it ferments：Containing after the seed liquor that step (1) culture obtains has been sterilized by the inoculum concentration access of 1% weight ratio
There are peptone 1-5% by weight percentage, yeast extract 0.5-5%, glucose 1-10%, L-cysteine hydrochloride
0.01-1%, ammonium sulfate 0.1%, potassium dihydrogen phosphate 0.05-0.3%, dipotassium hydrogen phosphate 0.05-0.3%, manganese sulfate 0.01-
0.05%th, magnesium sulfate 0.001-0.005% and calcium chloride 0.001-0.005%, remaining for deionized water fluid nutrient medium in,
Through 35 DEG C~37 DEG C stand anaerobic fermentations for 24 hours~48h；
(3) thalline is collected：The bifidobacterium breve LJM-006 zymotic fluids that step (2) culture is obtained, 3000rpm centrifugations 5~
10min removes supernatant, obtains bacterium mud, and bacterial suspension is made in 2~3 washings of deionized water, again with 3000rpm from
5~10min of the heart removes supernatant fluid and obtains sediment；
(4) bacterium broken wall：By the sediment that step (3) obtains by 1: 5~1: 10 plus deionized water, under 150MPa pressure
Clasmatosis obtains breaking-wall cell product；
(5) finished product：The sediment obtained in step (4) is freeze-dried under vacuum negative pressure condition, you can obtain purity
Reach more than 95% dried powder to get to bifidobacterium breve active factors.
Bifidobacterium breve (Bifidobacterium breve) LJM-006 of the present invention, in preservation on October 28 in 2011
In China Committee for Culture Collection of Microorganisms's common micro-organisms center, it is referred to as CGMCC (addresses：Chaoyang District, Beijing City
The institute 3 of North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101) preservation, Classification And Nomenclature is bifidobacterium breve
(Bifidobacterium breve), deposit number are CGMCC No.5418.
Bifidobacterium breve active factors of the present invention are preparing answering for treatment Alzheimer disease drugs for medicament active composition
With, medicament is made with auxiliary material combination, as long as the medicament belongs to the protection model of the present invention for treating Alzheimer disease
It encloses.
Advantages of the present invention and effect：
The invention firstly discloses ginkolide B, bifidobacterium breve active factors to treat Alzheimer for key component
The purposes of disease, and the preparation method of bifidobacterium breve active factors is provided.
Specific embodiment
Embodiment 1
The present invention includes each component of following weight for treating the composition of Alzheimer disease：
| Composition | Weight (mg) |
| Ginkolide B | 88 |
| Bifidobacterium breve active factors | 6 |
| Glyceryl monostearate | 6 |
The preparation method of bifidobacterium breve active factors
(1) prepared by seed liquor：Picking single bacterium colony bifidobacterium breve (Bifidobacterium breve) LJM-006, preservation
Number is CGMCC No.5418, is inoculated in sterilized MRS fluid nutrient mediums, 37 DEG C, and Anaerobic culturel is for 24 hours to get to short pair
Discrimination bacillus LJM-006 seed liquors；
(2) it ferments：Containing after the seed liquor that step (1) culture obtains has been sterilized by the inoculum concentration access of 1% weight ratio
There is peptone 3%, yeast extract 2%, glucose 8%, L-cysteine hydrochloride 0.05%, sulphur by weight percentage
Sour ammonium 0.1%, potassium dihydrogen phosphate 0.1%, dipotassium hydrogen phosphate 0.1%, manganese sulfate 0.02%, magnesium sulfate 0.002% and calcium chloride
0.002%, remaining is in the fluid nutrient medium of deionized water, anaerobic fermentation is stood for 24 hours through 35 DEG C；
(3) thalline is collected：The bifidobacterium breve LJM-006 zymotic fluids that step (1) culture is obtained, 3000rpm centrifugations
6min removes supernatant, obtains bacterium mud, and 3 washings of deionized water are made bacterial suspension, are centrifuged again with 3000rpm
9min removes supernatant fluid and obtains sediment；
(4) bacterium broken wall：By the sediment that step (2) obtains by 1: 5 plus deionized water, cell is broken under 150MPa pressure
It is broken, obtain breaking-wall cell product；
(5) finished product is lyophilized：The sediment obtained in step (3) is freeze-dried under vacuum negative pressure condition, you can obtain
Purity reaches more than 95% dried powder to get to bifidobacterium breve active factors.
Described bifidobacterium breve (Bifidobacterium breve) LJM-006, was deposited on October 28th, 2011
China Committee for Culture Collection of Microorganisms's common micro-organisms center is referred to as CGMCC (addresses：Chaoyang District, Beijing City north
The institute 3 of occasion West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101) preservation, Classification And Nomenclature is bifidobacterium breve
(Bifidobacterium breve), deposit number are CGMCC No.5418.
Ginkolide B standard items are purchased from Yuan Ye biotech firms.
Embodiment 2
The present invention includes each component of following weight for treating Alzheimer disease composition：
| Composition | Weight (mg) |
| Ginkolide B | 80 |
| Bifidobacterium breve active factors | 10 |
| Glyceryl monostearate | 10 |
The wherein preparation method of bifidobacterium breve active factors
(1) prepared by seed liquor：Picking single bacterium colony bifidobacterium breve (Bifidobacterium breve) LJM-006, preservation
Number is CGMCC No.5418, is inoculated in sterilized MRS fluid nutrient mediums, 37 DEG C, and Anaerobic culturel is for 24 hours to get to short pair
Discrimination bacillus LJM-006 seed liquors；
(2) it ferments：Containing after the seed liquor that step (1) culture obtains has been sterilized by the inoculum concentration access of 1% weight ratio
There is peptone 5%, yeast extract 1%, glucose 7%, L-cysteine hydrochloride 0.5%, sulphur by weight percentage
Sour ammonium 0.1%, potassium dihydrogen phosphate 0.2%, dipotassium hydrogen phosphate 0.05%, manganese sulfate 0.05%, magnesium sulfate 0.003% and calcium chloride
0.003%, remaining is in the fluid nutrient medium of deionized water, anaerobic fermentation is stood for 24 hours through 37 DEG C；
(3) thalline is collected：The bifidobacterium breve LJM-006 zymotic fluids that step (1) culture is obtained, 3000rpm centrifugations
5min removes supernatant, obtains bacterium mud, and 3 washings of deionized water are made bacterial suspension, are centrifuged again with 3000rpm
10min removes supernatant fluid and obtains sediment；
(4) bacterium broken wall：By the sediment that step (2) obtains by 1: 5 plus deionized water, cell is broken under 150MPa pressure
It is broken, obtain breaking-wall cell product；
(5) finished product is lyophilized：The sediment obtained in step (3) is freeze-dried under vacuum negative pressure condition, you can obtain
Purity reaches more than 95% dried powder to get to bifidobacterium breve active factors.
Described bifidobacterium breve (Bifidobacterium breve) LJM-006, was deposited on October 28th, 2011
China Committee for Culture Collection of Microorganisms's common micro-organisms center is referred to as CGMCC (addresses：Chaoyang District, Beijing City north
The institute 3 of occasion West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101) preservation, Classification And Nomenclature is bifidobacterium breve
(Bifidobacterium breve), deposit number are CGMCC No.5418.It is public that ginkolide B standard items are purchased from source leaf biology
Department.
3 zoopery of embodiment
Ability of learning and memory test is carried out using Morris water mazes, is tested including constant-bearing navigation and space exploration.Using
ELISA method detects the inflammatory factors levels such as IL-1 β, IL-6, TNF-α of brain tissue, and kit specification is pressed in concrete operations.
Experimental result is shown：Alzheimer disease model mouse occurs under apparent ability of learning and memory and judgement
Drop；1 treatment group of embodiment is obviously improved the learning memory disorder of Alzheimer disease model mouse, and constant-bearing navigation achievement (is escaped
Incubation period) it is obviously shortened, with model group than significant difference (P ＜ 0.01).Ginkolide B processing alzheimer ' is used alone
Silent disease model mouse has the effect for the learning memory disorder for improving Alzheimer disease model mouse, but effect is weaker than this
Inventive embodiments 1.The above result shows that the present invention has the learning and memory function for being obviously improved Alzheimer disease mouse.
The inflammatory factors such as Alzheimer disease model mouse IL-1 β, IL-6, TNF-α are horizontal significantly raised；Embodiment 1 is controlled
Treatment group is substantially reduced IL-1 β, IL-6, TNF-α level (P ＜ 0.05), and ginkolide B processing Alzheimer disease mould is used alone
Type mouse has the effect for the learning memory disorder for improving Alzheimer disease model mouse, but effect is weaker than the present invention in fact
Apply example 1.
Conclusion：
Compared with prior art, the present invention start prevention Alzheimer disease new model, have developed a kind of good effect,
The drug of prevention Alzheimer disease having no toxic side effect, proves, hence it is evident that it is small to improve Alzheimer disease through animal experiment study
The ability of learning and memory of mouse effectively reduces IL-1 β, IL-6, TNF-α level, mitigates the inflammatory reaction of microglia.
In conclusion only the preferred embodiments of the invention, not it is used for limiting the range of implementation of the invention.It is i.e. all
The equivalent changes and modifications done according to scope of the present invention patent are all that the scope of the claims of the present invention is covered.
Claims (1)
1. a kind of for treating the composition of Alzheimer disease, the composition include ginkolide B, bifidobacterium breve activity because
Son and glyceryl monostearate, each component weight ratio are：
Ginkolide B：85-100 parts by weight；
Bifidobacterium breve active factors：5-10 parts by weight；
Glyceryl monostearate：5-15 parts by weight；
The preparation method of the bifidobacterium breve active factors is as follows：
(1) prepared by seed liquor：Picking single bacterium colony bifidobacterium breve (Bifidobacterium breve) LJM-006, deposit number
It for CGMCC No.5418, is inoculated in sterilized MRS fluid nutrient mediums, 37 DEG C, Anaerobic culturel is for 24 hours to get to short bifid bar
Bacterium LJM-006 seed liquors；
(2) it ferments：After step (1) the obtained seed liquor of culture has been sterilized by the inoculum concentration access of 1% weight ratio contain with
The peptone 1-5% of weight percent meter, yeast extract 0.5-5%, glucose 1-10%, L-cysteine hydrochloride
0.01-1%, ammonium sulfate 0.1%, potassium dihydrogen phosphate 0.05-0.3%, dipotassium hydrogen phosphate 0.05-0.3%, manganese sulfate 0.01-
0.05%th, magnesium sulfate 0.001-0.005% and calcium chloride 0.001-0.005%, remaining for deionized water fluid nutrient medium in,
Through 35 DEG C~37 DEG C stand anaerobic fermentations for 24 hours~48h；
(3) thalline is collected：The bifidobacterium breve LJM-006 zymotic fluids that step (2) culture is obtained, 3000rpm centrifugations 5~
10min removes supernatant, obtains bacterium mud, and bacterial suspension is made in 2~3 washings of deionized water, again with 3000rpm from
5~10min of the heart removes supernatant fluid and obtains sediment；
(4) bacterium broken wall：By the sediment that step (3) obtains by 1: 5~1: 10 plus deionized water, the cell under 150MPa pressure
It is broken, obtain breaking-wall cell product；
(5) finished product：The sediment obtained in step (4) is freeze-dried under vacuum negative pressure condition, you can obtain purity and reach
More than 95% dried powder is to get to bifidobacterium breve active factors.
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| CN201711500596.1A CN108125988A (en) | 2017-12-31 | 2017-12-31 | Ginkolide B is in the application that microglia inflammatory reaction is inhibited to mitigate Alzheimer disease symptoms |
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|---|---|---|---|
| CN201711500596.1A CN108125988A (en) | 2017-12-31 | 2017-12-31 | Ginkolide B is in the application that microglia inflammatory reaction is inhibited to mitigate Alzheimer disease symptoms |
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| CN201711500596.1A Pending CN108125988A (en) | 2017-12-31 | 2017-12-31 | Ginkolide B is in the application that microglia inflammatory reaction is inhibited to mitigate Alzheimer disease symptoms |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111172074A (en) * | 2020-01-21 | 2020-05-19 | 北京科拓恒通生物技术股份有限公司 | Bifidobacterium lactis Probio-M8 capable of relieving and improving Alzheimer symptoms and application |
| CN113332285A (en) * | 2021-07-26 | 2021-09-03 | 大理大学 | Application of BMS470539 in preparation of medicine for treating Alzheimer's disease |
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Application publication date: 20180608 |