CN108498523A - The preparation method and applications of the ganglioside derivative of the chain containing unsaturated fatty acid - Google Patents
The preparation method and applications of the ganglioside derivative of the chain containing unsaturated fatty acid Download PDFInfo
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- CN108498523A CN108498523A CN201710102185.0A CN201710102185A CN108498523A CN 108498523 A CN108498523 A CN 108498523A CN 201710102185 A CN201710102185 A CN 201710102185A CN 108498523 A CN108498523 A CN 108498523A
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- Prior art keywords
- fatty acid
- carbon atom
- unsaturated fatty
- ganglioside derivative
- neurite outgrowth
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- 230000002441 reversible effect Effects 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 125000005471 saturated fatty acid group Chemical group 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- WWUZIQQURGPMPG-KRWOKUGFSA-N sphingosine Chemical compound CCCCCCCCCCCCC\C=C\[C@@H](O)[C@@H](N)CO WWUZIQQURGPMPG-KRWOKUGFSA-N 0.000 description 1
- 150000003410 sphingosines Chemical class 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 231100000816 toxic dose Toxicity 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000000472 traumatic effect Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Classifications
-
- 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
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
-
- 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
Abstract
Application of the ganglioside derivative for containing unsaturated fatty acid chain the present invention relates to one group in promoting Neurite Outgrowth.The ganglioside derivative library of unsaturated fatty acid chain is carried by sphingolipid ceramide N removal of acylation enzyme enzymatic clarifications; in conjunction with the cell model of classical rush Neurite Outgrowth; it was found that the ganglioside derivative with unsaturated fatty acid chain has the biological activity of the rush Neurite Outgrowth significantly improved, and the different biological activities for also promoting Neurite Outgrowth to it of the carbochain quantity of gangliosides sphingol base have a significant impact.
Description
Technical field
The invention belongs to biotechnologies, more particularly it relates to one group of nerve for containing unsaturated fatty acid
The high efficiency preparation method for saving glycosides fat derivative and the application on promoting Neurite Outgrowth.
Background technology
Gangliosides are a kind of glycosyl sphingolipids containing sialic acid residues, are the typical component parts of nerve cell plasma membrane,
There is important biological function.Ganglioside GM1 is the representational important gangliosides of one of which, is had
Multiple neurotrophic effect can promote cell differentiation, induction of neurite to generate, protect neuronal damage, adjusting nerve can
Plasticity.Clinically, GM1 have been widely used for treatment vascular or traumatic central nervous system damage, Parkinson's disease and Ah
Alzheimer's disease etc..
Gangliosides largely exist in mammalian brain, wherein GM1, GD1a, GD1b and GT1b etc. be mainly at
Part, the 97% of total gangliosides is accounted for about in human brain.Such compound structure is complicated, obtains these nervons at present
Main method is extracted from natural biological, and in addition to mammalian tissues, gangliosides are also distributed widely in other vertebras
In animal, plant, eukaryon and prokaryotic micro-organisms, type is various, various structures.Have and be much reported in the echinoderm of ocean,
Separation and Extraction is such as isolated in sea cucumber and starfish a variety of neural with promoting to a variety of gangliosides with medical active
The gangliosides such as the gangliosides of enation, wherein SJG-2, LLG-5, LLG-3 and GAA-7 are tested in vitro shows it
With rush Neurite Outgrowth more higher than GM1 activity.However many gangliosides content in biology is very low, source also has
Limit, and there may be microheterogeneities for the gangliosides naturally extracted, and the problems such as by pathogen contamination.Therefore,
The method by naturally extracting is difficult to extract and screen from natural gangliosides various in style with specific medicinal work
The substance of property.
The fully synthetic complicated nervon of chemistry is especially difficult, and be related to the protection of many genes, deprotection with
And stereoselectivity and etc., if the full chemistry of LLG-3 synthesizes, for needing the reaction of 30 steps, and in chemical method transformation process easily
By-product is generated, yield is relatively low, and needs the organic solvent used a large amount of highly basic and polluted the environment, therefore is also difficult to
The gangliosides with medical active are obtained and screened by the fully synthetic method of chemistry.
More and more evidences show the difference of the fatty acid chain in gangliosides ceramide group to its biology
Activity has important influence, especially contains unsaturated fatty acid chain, can significantly change the structure of gangliosides, it is also possible to band
To have the substance of different medical values.Utilize sphingolipid ceramide N removal of acylation enzyme (Sphingolipid ceramide N-
Deacylase, SCDase) catalysis characteristics, the fatty acid chain of gangliosides can be replaced, establish containing unsaturation
The ganglioside derivative library of fatty acid chain.However, due to SCDase autocatalysis reversible reactions, synthetic reaction is often resulted in
Conversion ratio it is not high.
Neurite Outgrowth is Neural Differentiation and a newer critical process, to because of injured or neurodegenerative disease
Caused nervous function is impaired to have repair.Some researches show that Ganglioside GM1s to have Neurite Outgrowth activity,
It can promote neural source cell enation.The fatty acid chain of gangliosides with Lipid Rafts and embrane-associated protein by interacting
The biological activity of gangliosides is greatly influenced, and there is presently no to the gangliosides containing unsaturated fatty acid chain
Derivative carries out system research.Whether unsaturated fatty acid chain can influence the rush Neurite Outgrowth activity of gangliosides, mesh
Preceding relevant research not yet is reported.
It would therefore be highly desirable to by a kind of easy, efficient method to obtain the gangliosides containing unsaturated fatty acid chain
Derivates library, and further effect of the research related derivatives in Neurite Outgrowth on this basis.
Invention content
The present invention obtains a series of gangliosides with unsaturated fatty acid chain using SCDase enzymatic clarification systems
Derivative, and be applied in the biological function screening for promoting Neurite Outgrowth, the study found that carrying unsaturated fat
The ganglioside derivative of sour chain contributes to the growth and development of nerve cell.
In the first aspect of the present invention, a kind of application of ganglioside derivative in promoting Neurite Outgrowth is provided,
The gangliosides are made of glycosyl, sphingol base and fatty acid chain, wherein sphingol base and fatty acid chain with
Amido bond, which is connected, constitutes ceramide, and glycosyl is connected with ceramide with glycosidic bond, and the aliphatic acid is unsaturated fatty acid；
In a preferred embodiment, the aliphatic acid is monounsaturated fatty acids；
In another preferred embodiment, the aliphatic acid is polyunsaturated fatty acid；
The carbochain of the unsaturated fatty acid includes 12~24 carbon atoms, and unsaturated bond is double or triple bonds；
Preferably, the aliphatic acid is straight chain olefin(e) acid, selected from the aliphatic acid containing ten four carbon atoms, contains 16
The aliphatic acid of carbon atom, the aliphatic acid containing 18 carbon atoms or the aliphatic acid containing 22 carbon atoms；
Preferably, the olefin(e) acid has 1 to 6 unsaturated double-bond, includes at least one or more following double bond, choosing
From：
(1) double bond is located at from fatty acid chain methyl end between the 3rd carbon atom and the 4th carbon atom；Or
(2) double bond is located at from fatty acid chain methyl end between the 5th carbon atom and the 6th carbon atom；Or
(3) double bond is located at from fatty acid chain methyl end between the 6th carbon atom and the 7th carbon atom；Or
(4) double bond is located at from fatty acid chain methyl end between the 7th carbon atom and the 8th carbon atom；Or
(5) double bond is located at from fatty acid chain methyl end between the 9th carbon atom and the 10th carbon atom；Or
(6) double bond is located at from fatty acid chain methyl end between the 12nd carbon atom and the 13rd carbon atom；Or
(7) double bond is located at from fatty acid chain methyl end between the 15th carbon atom and the 16th carbon atom；Or
(8) double bond is located at from fatty acid chain methyl end between the 18th carbon atom and the 19th carbon atom.
Preferably, the olefin(e) acid is selected from ω -3, ω -6, ω -7, ω -9 unsaturated fatty acids；The ω carbon atoms are
Refer to a carbon atom farthest apart from carboxyl in straight chain olefin(e) acid；The ω -3, ω -6, ω -7, ω -9 unsaturated fatty acids refer to
Be to go out on third carbon atom, the 6th carbon atom, the 7th carbon atom and the 9th carbon atom from ω carbon atom numbers
Existing first double bond.
Preferably, the monounsaturated fatty acids is selected from cis- 9- tetradecenoic acids (C14:1, n-9), cis- 9- hexadecylenes
Acid (C16:1, n-9), oleic acid (C18:1,n-9)；The polyunsaturated fatty acid is selected from cis- 4,7,10,13,
16,19- docosahexaenoic acids (DHA)；
The glycosyl is oligosaccharyl, including 2~10 saccharide residues, and has at least one sialic acid residues；
Preferably the oligosaccharides is selected from single sialic acid lactose, three hexose of single sialic acid, monosialyl tetrahexose；
It is highly preferred that the glycosyl is monosialyl tetrahexose；
The carbochain of the sphingol base includes 12~24 carbon atoms, selected from sphingol type long chain base, dihydro sheath ammonia
Alcohol type long chain base or phytosphingosine type long chain base；
The sphingol type long chain base is the long-chain sphingol for including 1,3- dihydroxy -2- amino -4- alkene；
The dihydrosphingosine type long chain base is the long-chain sphingol for including 1,3- dihydroxy -2- amino alkanes；
The phytosphingosine type long chain base is the long-chain sphingol for including 1,3,4- trihydroxy -2- amino alkanes；
Preferably, the sphingol base is sphingol type long chain base, and the carbon atom number is 18 or 20, packet
Include 1,3- dihydroxy -2- amino -4- octadecylenes or 1,3- dihydroxy -2- amino -4- icosa alkenes.
It is highly preferred that the ganglioside derivative with unsaturated fatty acid chain is selected from cis- 9- tetradecenoic acids-
GM1, cis- palmitoleic acid-GM1, oleic acid-GM1 or cis- 4,7,10,13,16,19- docosahexaenoic acids-
GM1。
Application of the ganglioside derivative in promoting Neurite Outgrowth, to derive the gangliosides
Object is for the food of the diseases such as neurodegenerative disease, neurotrosis, CO2 laser weld or nerve regneration, pharmaceutical composition or system
Application in the preparation of agent；
The pharmaceutical composition or preparation refer to include with unsaturated fatty acid chain ganglioside derivative and
Pharmaceutically acceptable carrier.
The second aspect of the present invention provides a kind of side preparing the ganglioside derivative containing unsaturated fatty acid chain
Method, using the synthetic reaction system of sphingolipid ceramide N- removal of acylation enzymes, the reaction system includes (1) buffer system,
Control ph is in the range of 6.0~8.0；(2) hemolytic gangliosides；(3) unsaturated fatty acid；(4) unsaturated fat
Sour cosolvent；And (5) sphingolipid ceramide N- removal of acylation enzymes；
Wherein, the concentration ratio of the hemolytic gangliosides and the unsaturated fatty acid is 1:1~4；System is reacted
Temperature control at 25~38 DEG C, 12~24 hours reaction time；Metal ion and surface are not contained in the reaction system
Activating agent.
Preferably, the buffer system, which can be PBS buffer solution, HEPES buffer solution, Tris buffer solutions etc., to maintain
The buffer system that pH value of solution is stablized；The cosolvent of the unsaturated fatty acid is preferably 10% glycol dimethyl ether；It is described
Reaction temperature be preferably 37 DEG C；
Preferably, the reaction system is established in the buffer system of pH7.2~7.8；
It is highly preferred that the reaction system is established in the buffer system of pH 7.5.
Synthesis reaction system provided by the present invention optimizes on reaction condition compared with prior art, and reaction is made to walk
Rapid easier, especially when pH value maintains 7.5, sphingolipid ceramide N- removal of acylation enzymes have relatively high synthesis and live
Property and relatively low hydrolysing activity limit the metal ion for inhibiting synthetic reaction and surface and in optimizing reaction system
The addition of activating agent has efficiently controlled the compound direction of sphingolipid ceramide N- removal of acylation enzymes；Meanwhile it also being added in system
The cosolvent of unsaturated fatty acid dissolving, the cosolvent is promoted to be very easy to removal, do not interfere with the separation of subsequent compound
Purifying.Due to the step of not using surfactant, isolating and purifying operate it is simpler, avoid surfactant cleaning step
Suddenly, synthesis reaction solution can directly be purified on solid-phase extraction column (such as Waters, US Sep Pak tC18),
Significantly shorten purification step and purification time.Therefore, synthetic reaction system provided by the present invention is compared with the existing technology
It is more efficiently and easy.
The other aspects of the present invention are apparent to those skilled in the art due to this disclosure
's.
Description of the drawings
The structural schematic diagram of Fig. 1 Ganglioside GM1s.
The flow chart of Fig. 2 enzymatic clarification GM1 derivatives
The GM1 derivatives of 10 kinds of chains containing unsaturated fatty acid of Fig. 3 enzymatic clarifications
The rush Neurite Outgrowth expression activitiy of Fig. 4 .GM1 and its derivative.With the GM1 of 25 μM (A) or 50 μM (B) or its
Derivative stimulate Neuro2a cells for 24 hours after, count the ratio of the cell containing protrusion.
The difference photo of the derivative induced Neuro2a cell differentiations of GM1s of Fig. 5 with unsaturated fatty acid chain, scale are
100μm
Fig. 6 contain the GM1 derivatives of different unsaturated fatty acid chains and sphingol base to cellular neural enation
It influences.(A-B) ratio of the cell containing protrusion.(C-D) longest projection length.(E-F) protrusion number.
The cell toxicity test of GM1 derivatives of Fig. 7 with unsaturated fatty acid chain and different sphingol bases
Specific implementation mode
The present inventor utilizes the two-way catalysis of SCDase, and hemolytic neuromere is obtained being fully hydrolyzed gangliosides
On the basis of glycosides fat, synthetic reaction condition is advanced optimized, has synthesized a series of gangliosides with unsaturated fatty acid chain
Fat derivative in conjunction with the cell model of Neurite Outgrowth, filters out a batch and has and significantly improve rush neural process on this basis
Play the ganglioside derivative of growth activity.
Gangliosides of the present invention refer to that with Fig. 1 be exemplary, by sugar chain, sphingol base and aliphatic acid
Chain forms；The sugar chain is the oligosaccharides for including at least one sialic acid, as Fig. 1 GM1 be gangliosides Typical Representative,
Its sugar chain portion is monosialyl tetrahexose, and sphingosine base portion is divided into 1,3- dihydroxy -2- amino -4- octadecylenes, fat
Fat acid chain part is octadecanoid acid.
Ganglioside derivative of the present invention with unsaturated fatty acid chain can be synthesized with SCDase
Product can also be based on the lipid extracts obtained in animal or plant or to arise directly from chemically synthesized product.As above
The ganglioside derivative can be modified.By modifying the ganglioside derivative of the present invention, can improve
Bioactivity, application of the active raising suitable for the ganglioside derivative during promoting Neurite Outgrowth.
Effect of the ganglioside derivative of the present invention in promoting Neurite Outgrowth, can be single substance, also may be used
To be mixture, can also be used together with other lipids, the condition used should make other lipids not influence ganglioside
The biological activity of fat derivative.
The present invention also provides a kind of food that can promote Neurite Outgrowth, pharmaceutical composition or preparations, including carry
The ganglioside derivative of unsaturated fatty acid chain is as active ingredient.The pharmaceutical composition includes：Described carries
The ganglioside derivative and pharmaceutically acceptable carrier of unsaturated fatty acid chain.
Ganglioside derivative provided by the present invention with unsaturated fatty acid chain can be because of periphery or maincenter
In nervous system disorder, damage, neurodegenerative disease or CO2 laser weld, regenerative process, promote Neurite Outgrowth, accelerates prominent
It touches and is formed.
The ingredient of " pharmaceutically acceptable " is suitable for people and/or mammal and without excessively bad side reaction (such as poison
Property), i.e., with rational benefit/risk than substance.Term " pharmaceutically acceptable carrier " refers to for Therapeutic Administration
Carrier, including various excipient and diluent.The term refers to some such medicament carriers：Themselves is not necessary work
Property ingredient, and apply after there is no excessive toxicity.
Suitable pharmaceutically acceptable carrier is well known to those of ordinary skill in the art.In Remington ' s
It can be found about pharmaceutically acceptable carrier in Pharmaceutical Sciences (Mack Pub.Co., N.J.1991)
Absolutely prove.Pharmaceutically acceptable carrier can contain liquid in the composition, as water, phosphate buffer, ringer are molten
Liquid, physiological saline, balanced salt solution, glycerine or sorbierite etc..In addition, there is likely to be complementary substance in these carriers,
Such as lubricant, glidant, wetting agent or emulsifier, pH buffer substance and stabilizer, such as albumin.
When in use, it is by the gangliosides of the present invention with unsaturated fatty acid chain of safe and effective amount
Derivative is applied to mammal (such as people), and the wherein safe and effective amount is considered as the factors such as administration route, patient health situation,
Within the scope of these are all skilled practitioners technical ability.
The present invention also provides a kind of injection delivery device (such as pumping needle), medicine box or kit, including：It is described
Ganglioside derivative with unsaturated fatty acid chain or the pharmaceutical composition.
For the ease of clinical application, pharmaceutical composition of the invention may be embodied in injection delivery device (such as pumping needle)
In, in the injection delivery device, the pharmaceutical composition of single administration amount can be included.The injection administration
Device can be contained in medicine box, to facilitate storage, use.
In medicine box of the present invention or kit, it may also include operation instructions, so that those skilled in the art press
It is used according to correct mode.
Cis unsaturated fatty acid of the present invention, for the aliphatic acid with unsaturated bond, and hydrogen atom is distributed in not
The homonymy of saturated bond.By taking cis- 9- tetradecenoic acids as an example, refer to that the aliphatic acid forms the cis- alkene of straight chain by 14 carbon atoms
Acid, unsaturated bond is located between the 9th carbon atom and the 10th carbon atom, is abbreviated as (C14 from c-terminus:1, n-9),
Other unsaturated fatty acids and so on.
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.
The general explanation in the source of biochemical material of the present invention：
1, unsaturated fatty acid is purchased from lark prestige scientific and technological (Shanghai) and Aladdin reagent (Shanghai) respectively；Sep-Pak tC18
Cartridge is purchased from Waters, US, and agents useful for same is that analysis is pure.
2, Neuro2a cells are derived from Shanghai Communications University nano biological engineering in medicine research institute Jin Weilin reality teachers experiment
Room；DMEM culture mediums are purchased from Corning companies；Fetal calf serum (FBS) and pancreatin are purchased from Gibco；Nonessential amino acid and green chain
Mycin is (dual anti-) to be purchased from Invitrogen companies；MTT is purchased from Sigma-Aldrich (St.Louis, USA).
The enzymatic clarification in ganglioside derivative library of the embodiment 1. with unsaturated fatty acid chain
Since the rush Neurite Outgrowth effect of current Ganglioside GM1 is the most clear, we initially set up
The derivates library of GM1 with unsaturated fatty acid chain, specific implementation mode are as follows：
The route of the GM1 derivatives of enzymatic clarification chain containing unsaturated fatty acid, is shown in Fig. 2.
First, the sphingolipid ceramide N- removal of acylation enzyme needed for catalytic synthesis and synthesis GM1 derivatives institute are prepared
The substrate hemolytic Ganglioside GM1 (Lyso-GM1) needed, specific step is according to the patent (application applied before us
Number：CN201410508221.X) and deliver article (J Lipid Res, 2015,56:1836-1842) prepared.
Synthetic reaction system：Reaction system is the second for containing about 10% in the Tris-HCl (pH 7.5) of the 50mM of 20ml
Glycol dimethyl ether (DME), the unsaturated fatty acid of the lyso-GM1 of about 20mg, 1.5mM.Into reaction system, every 8-12h plus
Enter the SCDase of the 1mg/ml of 1ml, whether complete coreaction for 24 hours, detects reaction by TLC.The solvent of TLC be chloroform,
Methanol, 0.02%CaCl2(5:4:1), colour reagent is CAM reagents (10g/L cerous sulfates, 50g/L ammonium molybdate tetrahydrates, 10% sulphur
Acid), the conversion ratio of synthetic reaction is 60-90%.
The GM1 derivatives of the 10 kinds of chains containing unsaturated fatty acid obtained in above-mentioned synthetic system, corresponding unsaturation
Fatty acid chain is respectively：Cis- linderic acid (C12:1, n-5), cis- 9- tetradecenoic acids (C14:1, n-9), cis- 11- bis-
Ten carbon enoic acid (C20:1, n-11) and cis- 4,7,10,13,16,19-docosahexaenoic acid (DHA).Cis- palmitoleic acid
(C16:1, n-9), trans- 2- gaidic acids (2HA) and cis-5,8,11,14,17-Eicosapentaenoic acid (EPA), cis- 9- ten
Eight carbon enoic acid (C18:1, n-9), cis-13-docosenoic acid (C22:1, n-13), cis-15-tetracosenoic acid (C24:1,
N-15), structure is shown in Fig. 3.
Other contain the ganglioside derivative of unsaturated fatty acid chain also according to method enzymatic clarification shown in Fig. 2.
Different gangliosides are hydrolyzed with SCDase prepare haemolysis gangliosides first.In 100ml reaction systems (pH 5.8), contain
The surfactant TDC of 0.4-0.8% (w/v)), the enzyme solutions of 10 μ l and the different neuromeres of final concentration of 0.2-10mM
The metallic ions Ca of glycosides fat (GM1, GM2, GM3, GD1, GD2, GD3, GT1, GT2, GT3 etc.), 100mM2+, reaction mixture is in 37
DEG C be incubated 12h, be fully hydrolyzed gangliosides, the hydrolysis degree of gangliosides detected by TLC.The solvent of TLC is
Chloroform, methanol, 0.02%CaCl2(5:4:1), colour reagent be CAM reagents (10g/L cerous sulfates, 50g/L ammonium molybdate tetrahydrates,
10% sulfuric acid).
Then it is reacted with unsaturated fatty acid with the haemolysis gangliosides prepared, anamorphic zone is not under the catalysis of SCDase
The ganglioside derivative of saturated fatty acid chain.Reaction system is to contain in the PBS (pH6.0~8.0) of the 50mM of 40ml
The haemolysis gangliosides of about 10% DME, about 40mg, the unsaturated fatty acid of 1~4mM.Into reaction system, every 8-
The SCDase of the 1mg/ml of 12h additions 1ml, coreaction 12~for 24 hours, it is whether complete that reaction is detected by TLC.The expansion of TLC
Agent is chloroform, methanol, 0.02%CaCl2(5:4:1), colour reagent is CAM reagents (10g/L cerous sulfates, tetra- water molybdic acids of 50g/L
Ammonium, 10% sulfuric acid)
Used unsaturated fatty acid can select olefin(e) acid or acetylenic acid, including ω -3, ω -6, ω -7, ω -9 unsaturated lipid
Fat acid.
Embodiment 2：Ganglioside derivative library with unsaturated fatty acid chain isolates and purifies
After completing catalytic synthesis, need to be isolated and purified out from reaction mixture with unsaturated fatty acid chain nerve
Save the derivative of glycosides fat.Here use Waters companies Sep-Pak tC18 solid-phase extraction columns (pillar of 1 500mg, 1
The pillar of 5g) it is isolated and purified.Due to not containing surfactant, synthesis reaction solution can be carried out directly in solid-phase extraction column
Purifying：Reaction mixture is first flowed through into a small Sep-Pak tC18 solid-phase extraction column (500mg), then liquid stream will be flowed through and cross one
A big Sep-Pak tC18 solid-phase extraction columns (5g), are washed with water 50ml, then wash 50ml with 50% methanol, finally with 100%
Methanol wash lower derivative, rotary evaporation is concentrated and is lyophilized.Alternatively, it is also possible to which reaction mixture is first flowed through a big Sep-
Pak tC18 solid-phase extraction columns (5g), are washed with water, and 50% methanol is washed, then wash lower mixture with the methanol of 90-100%, rotation
It is concentrated by evaporation and is lyophilized, be re-dissolved in water, cross a small Sep-Pak tC18 solid-phase extraction column (500mg), be washed with water, flow through liquid
As pure GM1 derivatives.Experiment is found, if there is organic solvent such as methanol in solution, can increase the hanging column of GM1 derivatives
Carrying capacity, GM1 derivatives can be made to lack hanging column to greatest extent so being re-dissolved in water with later approach, after freeze-drying, to increase
Add its yield.By the method for the above-mentioned purifying GM1 derivatives groped, 10 kinds of GM1 derivatives of synthesis easily purify
Out, purification yield between 16-67% (Fig. 3).
The derivative of gangliosides after each isolating and purifying is with HPLC-MS and high resolution mass spec (HRMS) come really
Recognize.Since gangliosides contain the sphingol base of two amount of carbon atom of difference, respectively C18- sphingols base (d18)
With C20- sphingols base (d20).Thus in each ganglioside derivative of synthesis, also contain both sphingosines
Base.
Embodiment 3：Ganglioside derivative with unsaturated fatty acid chain promotees the active measurement of Neurite Outgrowth
Neuro2a cells with 10% degrees of fusion kind in 24 orifice plates, culture for 24 hours after, be added prescribed concentration GM1 or
Its derivative, and after continuing culture for 24 hours, take a picture under phase contrast microscope, with Image Pro-Plus softwares number at least 300
Cells/well counts the ratio of the cell containing protrusion.In this experiment, projection length is more than the cell definition of 1 times of cell born of the same parents length
For the cell containing protrusion.In addition, the longest projection length of cell and the statistics of protrusion number have the thin of protrusion for 30 to minority
Born of the same parents.For the difference between more each experimental group, each experimental group is carried out with SPSS softwares (IBM, Chicago, IL, USA) independent
Sample t-test, * P<0.05, * * P<0.01, * * * P<0.001.
As shown in Figures 4 and 5, compared to GM1, derivative C14:1-GM1、C16:1-GM1、C18:1-GM1 and DHA-GM1
The Neuro2a cells of processing, the cell containing protrusion significantly increase, especially C16:1-GM1 and C18:1-GM1, processing
The ratio for the cell that cell contains protrusion is respectively：23.6% and 23.2%, and the cell of corresponding GM1 processing, contain protrusion
Cell proportion be 9.2%.In addition, the activity of DHA-GM1 is also higher, the cell proportion that the cell of processing contains protrusion is
13.9%.This result shows that the GM1 containing different unsaturated fatty acid chains has different rush Neurite Outgrowth activity simultaneously
(Fig. 4, Fig. 5) wherein C16:1-GM1 and C18:1-GM1 has rush enation more higher than GM1 activity.
The fatty acid chain of other gangliosides (such as GD1a, GD1b, GT1b etc.) transform as with ω -3, ω -6,
ω -7, ω -9 unsaturated fatty acid chain, such as cis- 11- tetradecenoic acids (C16:1, n-11), cis- 7- gaidic acids (C16:1,n-7)
And 13, when 16,19- docosatrienoic acid, the rush Neurite Outgrowth activity of opposite enhancing can also be shown.
Ganglioside derivative of the embodiment 4 containing different sphingol bases promotees the active influence of Neurite Outgrowth
According to embodiment 2 as a result, gm1 gangliosidosis includes that there are two types of sphingol bases, therefore, we further exist
Isolate GM1, C16:1-GM1 and C18:In 1-GM1 after each component, it is dense at 25 μM and 50 μM that each component is determined respectively
Under degree, their rush Neurite Outgrowth activity.As shown in Fig. 6 results, each component shows different rush nervous process lifes
It is long acting, even if fatty acid chain is identical, if sphingol base is different, promotees Neurite Outgrowth activity and also differ, including
Two component GM1 (d18) of GM1 and GM1 (d20), the activity of GM1 (d18) whether in 25 μM or 50 μM, compare GM1
(d20) height.It is consistent with the result of front, C16:1-GM1 and C18:Two component Cs 16 in 1-GM1:1-GM1(d18)、
C16:1-GM1(d20)、C18:1-GM1 (d18) and C18:1-GM1 (d20), activity are above two components (Fig. 6) of GM1,
Especially C16:The Neuro2a cells of 1-GM1 (d18), processing contain the ratio highest of protrusion cell.Each group is counted simultaneously
The longest projection length and protrusion number of part treated Neuro2a cells containing protrusion.As shown in fig. 6, two with GM1
A component is compared, C16:The cell of 1-GM1 (d18) processing, longest projection length and protrusion number are also obviously than GM1 (d18)
It is more with the cell of GM1 (d20) processing.
Embodiment 5 carries the cell toxicity test of unsaturated fatty acid ganglioside derivative
GM1(d18)、GM1(d20)、C16:1-GM1(d18)、C16:1-GM1(d20)、C18:1-GM1 (d18) and C18:
1-GM1 (d20) detects the toxicity of Neuro2a cells with mtt assay.As shown in fig. 7, C16:1-GM1(d18)、C16:1-GM1
(d20)、C18:1-GM1(d18)、C18:The half cell of 1-GM1 (d20), GM1 (d18) and GM1 (d20) to Neuro2a cells
Toxic concentration (CC50) is respectively 61.56,349.8,285.8,2420,2447 and 3645 μM.
Ganglioside derivative of the embodiment 6 with unsaturated fatty acid promotes in nerve growth development and plays important tune
The expression of the albumen of control effect
The reason of ganglioside derivative increased activity for being comparative analysis with unsaturated fatty acid, we are to it
Transcription group sequencing analysis is carried out.It is with final concentration of 25 μM of C16 first:1-GM1 (d18) and GM1 (d18) processing
Neuro2a cells and control Neuro2a cells, compare GM1 processing group and control group, risen in transcription negative regulation because
Son, such as DNA combinations repressor 2 and 3 (Inhibitor of DNA-binding protein 2and 3, Id2and Id3) table
Up to downward, and the two Transcription inhibition play negative regulation in Neural Differentiation.Negative regulation is played in Neural Differentiation simultaneously
The related enhancer 5 (Hairy and enhancer of split 5, Hes5) of transcription factor hair-like division, which is expressed, lowers, research
Show to inhibit the expression of the albumen can promote Neural Differentiation.And the transcription factor with important regulating and controlling effect in terms of nerve growth
Early growth reactive protein 1 (Early growth response protein 1, Egr1) and Kruppel like factors 7
(Kruppel-like factor 7, Klf7) up-regulated expression.Wherein, Egr1 is extracellular signal-regulated kinase
The downstream gene of (Etracellular regulated kinase, Erk), research shows that it can promote Neurite Outgrowth, and
Klf7 has important regulating and controlling effect to the expression of Neural Differentiation and TrkA genes.C16:1-GM1(d18:1) neural life is further promoted
The expression that the albumen of important regulating and controlling effect is played in long development, such as 7 (Ephrin type-A of memebrane protein Ephrin A receptors
Receptor 7, Epha7) and cytokinesis pay protein 10 (Dedicator of cytokinesis, Dock10).
Wherein, Epha7 can mediate the activation of Erk；Dock10 can active cell division control protein 42 (Cell division
Control protein 42, Cdc42) albumen, which can promote Neuronal processes to grow.
Other, which are observed, promotees the ganglioside derivative with unsaturated fatty acid chain that Neurite Outgrowth activity improves
Protein expression have also discovered similar regulation rule, further demonstrate the gangliosides with unsaturated fatty acid chain and spread out
Effect of the biology in promoting nervous system development.
All references mentioned in the present invention is incorporated herein by reference, independent just as each document
It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can
To be made various changes or modifications to the present invention, such equivalent forms equally fall within model defined by the application the appended claims
It encloses.
Claims (10)
1. a kind of application of ganglioside derivative in promoting Neurite Outgrowth, the ganglioside derivative is by sugar
Base, sphingol base and fatty acid chain composition, wherein sphingol base, which are connected with fatty acid chain with amido bond, is constituted neural acyl
Amine, glycosyl are connected with ceramide with glycosidic bond, which is characterized in that the aliphatic acid is unsaturated fatty acid, selected from containing
The aliphatic acid of ten four carbon atoms, the aliphatic acid containing 16 carbon atoms, the aliphatic acid containing 18 carbon atoms contain
There is the aliphatic acid of 22 carbon atoms.
2. a kind of application of the ganglioside derivative according to claim 1 in promoting Neurite Outgrowth, feature
Be, the unsaturated fatty acid be straight chain olefin(e) acid, the olefin(e) acid have 1 to 6 unsaturated double-bond, include with down toward
Few one or more double bond, is selected from：
(1) double bond is located at from fatty acid chain methyl end between the 3rd carbon atom and the 4th carbon atom；Or
(2) double bond is located at from fatty acid chain methyl end between the 5th carbon atom and the 6th carbon atom；Or
(3) double bond is located at from fatty acid chain methyl end between the 6th carbon atom and the 7th carbon atom；Or
(4) double bond is located at from fatty acid chain methyl end between the 7th carbon atom and the 8th carbon atom；Or
(5) double bond is located at from fatty acid chain methyl end between the 9th carbon atom and the 10th carbon atom；Or
(6) double bond is located at from fatty acid chain methyl end between the 12nd carbon atom and the 13rd carbon atom；Or
(7) double bond is located at from fatty acid chain methyl end between the 15th carbon atom and the 16th carbon atom；Or
(8) double bond is located at from fatty acid chain methyl end between the 18th carbon atom and the 19th carbon atom.
3. a kind of application of the ganglioside derivative according to claim 2 in promoting Neurite Outgrowth, feature
It is, the unsaturated fatty acid is selected from:Cis- 9- tetradecenoic acids (C14:1, n-9), cis- palmitoleic acid (C16:1,n-
9), oleic acid (C18:1, n-9) or cis- 4,7,10,13,16,19- docosahexaenoic acids (DHA).
4. a kind of application of the ganglioside derivative according to claim 1 in promoting Neurite Outgrowth, feature
It is, the glycosyl is oligosaccharides, including 2~10 saccharide residues, and there are at least one sialic acid residues.
5. a kind of application of the ganglioside derivative according to claim 4 in promoting Neurite Outgrowth, feature
It is, the oligosaccharides is selected from single sialic acid lactose, three hexose of single sialic acid or monosialyl tetrahexose.
6. a kind of application of the ganglioside derivative according to claim 1 in promoting Neurite Outgrowth, feature
It is, the sphingol base includes 12~24 carbon atoms, long selected from sphingol type long chain base, dihydrosphingosine type
Chain base or phytosphingosine type long chain base.
7. a kind of application of the ganglioside derivative according to claim 6 in promoting Neurite Outgrowth, feature
It is, the sphingol base is selected from 1,3- dihydroxy -2- amino -4- octadecylenes or 1,3- dihydroxy -2- amino -4- two
Ten alkene.
8. a kind of application of the ganglioside derivative according to claim 1 in promoting Neurite Outgrowth, feature
It is, the application is the ganglioside derivative for neurodegenerative disease, neurotrosis, CO2 laser weld or god
Application in the preparation of food, pharmaceutical composition or preparation through diseases such as regeneration.
9. a kind of preparation method of the ganglioside derivative containing unsaturated fatty acid chain, which is characterized in that utilize sphingolipid
The synthetic reaction system of ceramide N- removal of acylation enzymes, the reaction system include (1) buffer system, and control ph exists
In the range of 6.0~8.0；(2) hemolytic gangliosides；(3) unsaturated fatty acid；(4) unsaturated fatty acid cosolvent；With
And (5) sphingolipid ceramide N- removal of acylation enzymes；
Wherein, the concentration ratio of the hemolytic gangliosides and the unsaturated fatty acid is 1:1~4；The temperature of system reaction
Degree control is at 25~38 DEG C, 12~24 hours reaction time；Metal ion and surface-active are not contained in the reaction system
Agent.
10. a kind of preparation method of ganglioside derivative containing unsaturated fatty acid chain according to claim 9,
It is characterized in that：The reaction system is established in the buffer system of pH 7.5.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109161576A (en) * | 2018-09-26 | 2019-01-08 | 武汉中科光谷绿色生物技术有限公司 | Promote the method for fermentation of bacillus subtilis production N-acetyl-neuraminate |
| CN110464874A (en) * | 2019-08-30 | 2019-11-19 | 中国科学院深圳先进技术研究院 | A kind of polymer material and its preparation method and application with nerve fiber repairing activity |
| CN112961202A (en) * | 2021-03-15 | 2021-06-15 | 武汉糖智药业有限公司 | Preparation and purification method of monosialotetrahexosyl ganglioside |
| CN114085254A (en) * | 2020-08-24 | 2022-02-25 | 上海交通大学 | Ganglioside GM3 derivative and its preparing process and application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4476119A (en) * | 1981-08-04 | 1984-10-09 | Fidia S.P.A. | Method for preparing ganglioside derivatives and use thereof in pharmaceutical compositions |
| CN1052868A (en) * | 1989-11-14 | 1991-07-10 | 菲迪安股份公司 | Utilize natural Sphingolipids,sialo or derivatives thereof to stablize and keep the biologic activity of nerve growth factor |
| CN102202524A (en) * | 2008-01-28 | 2011-09-28 | 美赞臣营养品公司 | Nutritional composition containing dha, rumenic acid, and gangliosides |
| CN104293843A (en) * | 2014-09-28 | 2015-01-21 | 上海交通大学 | Method for preparing haemolytic sphingolipid by efficiently hydrolyzing sphingolipid |
-
2017
- 2017-02-24 CN CN201710102185.0A patent/CN108498523B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4476119A (en) * | 1981-08-04 | 1984-10-09 | Fidia S.P.A. | Method for preparing ganglioside derivatives and use thereof in pharmaceutical compositions |
| CN1052868A (en) * | 1989-11-14 | 1991-07-10 | 菲迪安股份公司 | Utilize natural Sphingolipids,sialo or derivatives thereof to stablize and keep the biologic activity of nerve growth factor |
| CN102202524A (en) * | 2008-01-28 | 2011-09-28 | 美赞臣营养品公司 | Nutritional composition containing dha, rumenic acid, and gangliosides |
| CN104293843A (en) * | 2014-09-28 | 2015-01-21 | 上海交通大学 | Method for preparing haemolytic sphingolipid by efficiently hydrolyzing sphingolipid |
Non-Patent Citations (2)
| Title |
|---|
| YUN-BIN HAN ET AL: "Comprehensive characterization of sphingolipid ceramide N-deacylase for the synthesis and fatty acid remodeling of glycosphingolipids", 《APPL MICROBIOL BIOTECHNOL》, 17 February 2015 (2015-02-17), pages 1 - 12 * |
| 王红 等: "神经节苷脂在神经再生中的作用", 《山西医科大学学报》, vol. 46, no. 09, 26 September 2015 (2015-09-26), pages 936 - 940 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109161576A (en) * | 2018-09-26 | 2019-01-08 | 武汉中科光谷绿色生物技术有限公司 | Promote the method for fermentation of bacillus subtilis production N-acetyl-neuraminate |
| CN110464874A (en) * | 2019-08-30 | 2019-11-19 | 中国科学院深圳先进技术研究院 | A kind of polymer material and its preparation method and application with nerve fiber repairing activity |
| CN110464874B (en) * | 2019-08-30 | 2021-11-05 | 中国科学院深圳先进技术研究院 | Polymer material with nerve tissue repair activity and preparation method and application thereof |
| CN114085254A (en) * | 2020-08-24 | 2022-02-25 | 上海交通大学 | Ganglioside GM3 derivative and its preparing process and application |
| CN114085254B (en) * | 2020-08-24 | 2023-10-20 | 上海交通大学 | Ganglioside GM3 derivative, and preparation method and application thereof |
| CN112961202A (en) * | 2021-03-15 | 2021-06-15 | 武汉糖智药业有限公司 | Preparation and purification method of monosialotetrahexosyl ganglioside |
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