CN1850972A - Sialic acid oligose-chitosan complex, and its preparing method and use - Google Patents
Sialic acid oligose-chitosan complex, and its preparing method and use Download PDFInfo
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Abstract
The invention provides the salivary acid few candy-shell polysaccharide compound, characterized in that the salivary acid candy can be combined in the shell polysaccharide to form the compound with the shape of the ion combining. The said salivary acid few candy is a or several kind of the below substance: the salivary acid, the ramification of the salivary acid, the candy chain with the salivary, the candy with ramification of the salivary acid, the compound of the functional group of the salivary acid or the ramification of the salivary acid. The invention also provides the producing method, the virus sorbent containing the compound and the antivirus drug, the sanitation article and the industrial product using the virus sorbent provided by the invention for the effective component.
Description
Technical field
The present invention relates to sialic acid, sialic acid derivative or contain the compound (below, it is generically and collectively referred to as sialyloligosaccharide) of above-mentioned substance, complex body, its manufacture method that forms through ionic bond with chitosan and the viral sorbent material that contains this complex body.Simultaneously, antiviral drug, sanitary product and mechanicals that to the invention still further relates to viral sorbent material of the present invention be effective constituent.
Background technology
Have in the prior art by organic synthesis and enzymatic synthetic method sialyloligosaccharide is incorporated into the report that chitosan forms complex body, but this complex body not only needs the purified sialyloligosaccharide, and all be by covalent sialyloligosaccharide to be incorporated on the polymer carrier, exist that synthesis procedure is numerous and diverse, cost is high and use the defective or the deficiency of poisonous and harmful reagent.Describe with regard to related notion and data-searching below.
Say so to the sialic acid narrow sense general name of acid sugar of acyl derivative of N-n acetylneuraminic acid n (Neu5Ac), N-hydroxyacetylneuraminic acid neuraminic acids such as (Neu5Gc).Sialic derivatives such as particularly sialic deaminizing thing, acetylate, hydrosulfate, methide, lactoyl thing or lactone organizer are also in the multiple existence of nature; and also has the multiple sialic acid derivative of artificial synthetic; in a broad sense, sialic acid also comprises above-mentioned sialic acid derivative.Therefore, in specification sheets of the present invention, sialyloligosaccharide is not the sialic acid of narrow sense, comprises sialic derivative yet, generally also comprises the compound that contains sialic acid or sialic acid derivative.
Above-mentioned sialyloligosaccharide is incorporated into the non-reduced end of sugar chain of glycoprotein, glycosamine, glycolipid matter, when being used for the expression of negative electric charge, be distributed in the cell surface of organism, taking on important biological function as the sugar of the sugar chain that constitutes glycoprotein, glycolipid matter.For example, the most occasions of tip at the sugar chain relevant with the stretching, extension of the transfer of cancer cells, inflammatory effect, neurocyte etc. exist sialic acid.
And, represent the infection of several viruses of influenza virus, be after there is sialic compound saccharic sugar chain of containing of target cell surface (acceptor) in specific molecular (ligand) identification by viral side, cause that specific absorption carries out.For example, certain influenza virus is because of having sialidase and erythrocyte agglutination element (homo agglutinin), so identification is infected by the 3 sugared structures that (N-n acetylneuraminic acid n α 2 → 3-D-semi-lactosi β 1 → 4-N-ethanoyl-D-グ glycosamine) constitutes at adventitia.
Simultaneously, chitosan be present in the first skin of crab, shrimp, krill and insects N-deacetylation N-ethanoyl-D-glycosamine with β-1,4 in conjunction with after the poly-glycosamine that obtains through deacetylation of the chitin that obtains.Chitosan is a straight chain shape polymer substance, and most amino uniformly-spaced exists on its straight chain, is the unique cationic polymer with excellent characteristic as crude substance.
Chitosan has various physico and reaches physiologically active in vivo and be applied to various purposes because of the special structure of its glycosaminoglycan.For example, immobilized enzyme all gets along with application in each field such as carrier, metal ion adsorbent, immunostimulant, trauma care promotor, artificial organ, antiseptic-germicides.Simultaneously, as long as chemically or biological ground beautify chitosan molecule, promptly, can give chitosan with new function (physico or biological intravital physiologically active), and can enlarge purposes hugely as by on chitosan molecule, importing other functional molecular or functional group.
In recent years, utilize virus that sialyloligosaccharide is had this characteristic of recognition capability, will contain that sialyloligosaccharide is fixed in suitable polymer such as chitosan and the exploitation progressively development of the antiviral material that constitutes.For example, relevant for making sialic acid and polystyrene or polyacrylamide bonded virus adsorbent (document 1, document 2, patent documentation 1), or sialyloligosaccharide is incorporated into the report of the complex body (patent documentation 2, patent documentation 3) etc. of chitosan by link molecule.In addition also relevant for making the l-asparagine mating type sugar-protein sugar chain that contains the aglycone part of being with formyl radical be incorporated into the report (with reference to patent documentation 2) of the chitosan that contains l-asparagine mating type sugar-protein sugar chain on the amino of chitosan.
But these all are to use refined in fact sialyloligosaccharide to prepare, and have beyond the unfavorable factor on the manufacturing cost, also have the narrow range of adsorbable virus, can only effectively catch the present situation of specific virus.And these conventional arts all are incorporated on the polymer carrier sialyloligosaccharide by covalent linkage, not only need a large amount of loaded down with trivial details synthesis procedures, also must use poisonous or environmentally harmful chemical reagent, organic solvent.Therefore, on aspect operability and manufacturing cost, have the disadvantageous picture, in the application of field of medicaments such as medicine, sanitary product, also be very limited.
[document 1] A.Tsuchida, K.Kobayasi, N.Matsubara, T.Muramatu, T.Suzuki, and Y.Suzuki; Glycoconjugate Journal 15,1047-1054 (1998).
[document 2] T.Furuike, S.Aiba, T.Suzuki, T.Takehashi, Y.Suzuki, K.Yamada, S-I.Nishimura; J.Chem.Soc.Perkin Trans.1,2000,3000-3005.
[patent documentation 1] Japan special permission Room, Japanese kokai publication hei 9-234317 communique.The contriver: rapids three heros in river, river are rapids excellently to be controlled, hillside plot with become.Applicant: NGK Insulators Ltd.Open day: on September 9th, 1997.
[patent documentation 2] Japan special permission Room, TOHKEMY 2003-301002 communique.Contriver: herd that the village is abundant, bear paddy English face, Yamamoto constitution two.Applicant: Inst Noguchi.Open day: on October 21st, 2003.
[patent documentation 3] Japan special permission Room, TOHKEMY 2005-247907 communique.Contriver: Li Xuebing, one youth of western village gentry.Applicant: one youth of western village gentry, The Hokkaido Electric Power Co., Inc., bioorganic chemistry institute of Co., Ltd..Open day: on September 15th, 2005
Summary of the invention
The present invention is directed to the defective or the deficiency that exist in the prior art, the sialic acid oligose-chitosan complex that a kind of preparation section is easy, need not to use harmful chemical reagent and multiple virus is had adsorptive power is provided.
The present invention also provides the preparation method of above-mentioned sialic acid oligose-chitosan complex.
The present invention also provides the viral sorbent material that contains above-mentioned sialic acid oligose-chitosan complex.
Antiviral drug, sanitary product or mechanicals that it is effective constituent that the present invention also provides with above-mentioned viral sorbent material.
Technical conceive of the present invention is, the anionic property by utilizing sialyloligosaccharide and the cationic of chitosan, make both be combined into sialic acid oligose-chitosan complex, thereby a kind of new, effective and broad-spectrum product is provided with the form of ionic bond.
Technical solution of the present invention is as follows:
Sialic acid oligose-chitosan complex is characterized in that: sialyloligosaccharide is incorporated into chitosan formation complex body with the form of ionic bond.
Described sialyloligosaccharide is one or more of following material: sialic acid, sialic acid derivative contains sialic sugar chain, contains the sugar chain of sialic acid derivative, the compound that contains as functional group and intramolecularly with sialic acid or sialic acid derivative.
Described sialyloligosaccharide derives from the colostrum of mammal.
Described sialyloligosaccharide is for being the sialyloligosaccharide of representative with the sialyl lactose.
The preparation method of above-mentioned sialic acid oligose-chitosan complex is characterized in that: utilize the cationic of the anionic property and the chitosan of sialyloligosaccharide, make sialyloligosaccharide and chitosan carry out ionic bond.
Described sialyloligosaccharide is from the isolating mixture as enriched material of natural goods.
Described mixture is that the sialyl lactose that obtains in the cow's milk is the mixture of the sialyloligosaccharide of representative.
The virus sorbent material is characterized in that, comprises above-mentioned sialic acid oligose-chitosan complex.
The medicine of antiviral usefulness, sanitary product or mechanicals is characterized in that, are effective constituent with above-mentioned viral sorbent material.
The present invention provides adsorbable a greater variety of virus, and does not have numerous and diverse operation and deleterious chemical reagent for solving above-mentioned problem in the past, can prepare and can be used as the viral adsorbent that medicine uses easily.
The present invention by make sialyloligosaccharide, particularly derive from the breast of natural materials, particularly cow's milk, more preferably the sialyloligosaccharide that contains sialyl lactose and have all structures of bovine coloctrum directly is incorporated into chitosan with the ionic bond form, thereby solve above-mentioned problem.
Promptly the present invention relates to a kind of sialic acid oligose-chitosan complex, it is conceived to sialic acid or contains the anionic property of sialic sugar chain (sialyloligosaccharide), and sialyloligosaccharide is adsorbed on the chitosan molecule chain of cationic with the ionic bond form.Particularly relate to sialyloligosaccharide, preferred sialyl lactose is more specifically said so and is derived from the sialyloligosaccharide of breast or its processed products, particularly derives from the sialyloligosaccharide of cow's milk and the ion complex body of chitosan.The invention provides simultaneously as being the viral adsorbent of effective constituent with above-mentioned chitosan complex, the virus of its effective Anti-virus agent or filter membrane, mouth mask etc. is adsorbed mechanicals.
In addition, the invention still further relates to the manufacture method of sialic acid oligose-chitosan complex, it contains sialic acid or contains the sialic acid sugar chain and chitosan carries out ionic bond for making.The viral sorbent material that particularly the present invention relates to the sialic acid oligose-chitosan complex that contains the invention described above is the medicine or the Industrial products (for example fiber etc.) of effective constituent.
In more detail, the present invention following (1)～(17) is described.
(1) sialic acid or contain sialic sugar chain and chitosan carries out the sialic acid oligose-chitosan complex of ionic bond.
(2) sialic acid oligose-chitosan complex described in above-mentioned (1), its sialic acid or to contain sialic sugar chain be sialyloligosaccharide.
(3) each described sialic acid oligose-chitosan complex in above-mentioned (1)～(2), its sialic acid or contain sialic sugar chain and derive from breast or its machining object.
(4) sialic acid oligose-chitosan complex described in above-mentioned (3), its breast is a cow's milk.
(5) sialic acid oligose-chitosan complex described in above-mentioned (4), its cow's milk is colostrum.
(6) sialic acid oligose-chitosan complex described in above-mentioned (1)～(5), the molecular weight of its chitosan is in 200～2,000,000 scope.
(7) sialic acid oligose-chitosan complex described in above-mentioned (1)～(5), the degree of deacetylation of its chitosan is in 100%～10% scope.
(8) make the method for sialic acid oligose-chitosan complex, it contains sialic acid or contains sialic sugar chain and chitosan carries out ionic bond for making.
(9) method described in above-mentioned (8), its sialic acid or to contain sialic sugar chain be sialyloligosaccharide or its mixture.
(10) method described in above-mentioned (9), its sialyloligosaccharide or its mixture derive from breast.
(11) method described in above-mentioned (10), its breast is cow's milk or its converted products.
(12) viral sorbent material, it contains each described sialic acid oligose-chitosan complex in above-mentioned (1)～(7).
(13) the viral sorbent material described in above-mentioned (12), its sialic acid oligose-chitosan complex contains sialic acid or contains the mixture of sialic sugar chain.
(14) medical and sanitary products or industrial fiber goods, it contains the viral sorbent material described in above-mentioned (12) or (13).
(15) medical and sanitary products described in above-mentioned (14), its medical and sanitary products is meant the manufacturing raw material of oral medicine, externally applied agent, mouth-washes or spray or these medicines.
(16) fibre product described in above-mentioned (14), its fibre product is meant processing raw material of filter membrane or mouth mask or these medicines.
(17) virus is caught and is used composition, and it contains each described chitosan complex or virus absorption carrier in above-mentioned (1)～(16).
Of the present invention being characterized as used sialic acid, sialic acid derivative or with its compound that contains as functional group and intramolecularly, and makes these compounds be adsorbed in chitosan formation complex body with the form of ionic bond.
" sialic acid " among the present invention not only comprises the sialic acid as the acyl derivative of neuraminic acid; the various derivatives that also comprise these materials; for example, the neuraminic acid derivatives that comprises deaminizing thing, acetylate, hydrosulfate, methide, lactoyl thing etc.In addition, sialyloligosaccharide of the present invention is meant and contains sialic carbohydrate more than a kind or 2 kinds, can exemplify to above-mentioned carboxylic acid group or contains monose, disaccharides or oligosaccharides or these sugared derivatives etc.Preferred sialyloligosaccharide is meant the carbohydrate with carboxylic acid group, for example sialyloligosaccharide.In the sialyloligosaccharide, a large amount of sialyl lactoses that distribute of occurring in nature also for preferred especially, but are not limited to this from the easy acquisition not only in the seizure and reactivity of virus.Though sialyloligosaccharide of the present invention can use the single material of chemically purifying, and considers from easy acquisition and price, also can use from the isolating mixture as enriched material of natural goods.By using such sialyloligosaccharide mixture, from adsorbing on the multiple virus simultaneously, effective viral sorbent material as viral adsorbent of the present invention, when being not target with specific virus, as sialyloligosaccharide of the present invention, preferably uses with the mixture of sialyl lactose particularly newborn from nature, that more preferably obtain the cow's milk as the sialyloligosaccharide of representative.
Sialic acid oligose-chitosan complex of the present invention is that the carboxyl of sialyloligosaccharide and the amino of chitosan molecule generate with ionic bond absorption.Specifically, be in appropriate solvent, to dissolve chitosan with solubilized chitosans such as acetic acid, add sialyloligosaccharide, after making the amino and the absorption of the carboxyl in the sugar chain of chitosan, through well-known treating processes such as dialysis,, obtain required complex body by carrying out lyophilize.
At this, the complex body of chitosan and sialyloligosaccharide can be by chitosan and sialic acid, or contains chitosan and sialic sugar chain is prepared respectively, but prepares after wishing to make the mixture of sialyloligosaccharide and chitosan reaction.Relevant mixture wishes to use breast, or is the sialyloligosaccharide of representative with the sialyl lactose from the breast, particularly bovine coloctrum of animal, for example ox, goat, sheep.
The Ruzhong is except that the neutral sugar chain with lactose etc., also have contain free sialic acid or sialic acid in conjunction with after sialic sugar chains such as sialyl lactose, contain particularly that sugared content is equivalent to 5%～20% sialyloligosaccharide in the bovine coloctrum, and about these diversified structure (Robert G.Jensen と Marvin P.Thompson volume, Handbook of MilkComposition, nineteen ninety-five, Academic Press periodical, ISBN:0123844304.Rectify in the island, Pu, dairy industry technology 53 volume 56-68 pages or leaves, 2003.Paul MacJarrow and Janneke van Amelsfort-Schoonbeek, International Dairy Journal, 14 volume 571-579 pages or leaves, 2004).
Usually, as everyone knows, virus is also different because of the different institute of its kind sialic sugar chain structure of the bonded of discern, so by using the sialyloligosaccharide from the breast with varied structure, preferred cow's milk to prepare viral seizure body, can catch a greater variety of viruses.Except that sialyloligosaccharide from cow's milk, use has the multiple sialyloligosaccharide of various different structures from all origin purified of the colostrum that contains the people, also can prepare chitosan complex of the present invention, but, use cow's milk, preferably use sialyloligosaccharide in the bovine coloctrum from considerations such as easy acquisition or purification operations.Contain than the poor sialyloligosaccharide of bovine coloctrum in the normal breast of ox (cow's milk), so preferably utilize than the sialyloligosaccharide in the bovine coloctrum of the easy acquisition of bovine coloctrum.Bovine coloctrum and cow's milk are except that as the lactogenesis, also as utilizations such as processed foods such as butter, cheese, formulas, modified milk, milk powder, skim-milk, cow initial milk protein matter, heath food, animal pharmaceuticals.In the processing and manufacturing process of these existing commodity in the machining object of cheese that generate, that comprise market sale etc., sialyloligosaccharide is arranged by the material of refining or the state that is concentrated, it is also preferably as raw material of sialyloligosaccharide of the present invention.
So the sialic acid oligose-chitosan complex of the present invention of preparation can be caught system as viral adsorption with virus separately with it and be become effective viral sorbent material, also can mix as viral adsorbent being prepared into effective virus absorption composition with various carriers.Viral sorbent material of the present invention comprises and contains sialic acid oligose-chitosan complex of the present invention separately, and as above-mentioned mixing with water equal solvent shape carrier afterwards or the absorption of the virus after dissolving composition.
Form the carrier that mixture is used with the viral sorbent material of invention, so long as the absorption of the deactivationization of virus or virus is not produced unfavorable, all unrestricted.For example can exemplify preparation material of solvent into water etc., filamentary material, Mierocrystalline cellulose etc. etc.In the purposes of the mixture that this type of sialic acid oligose-chitosan complex and carrier form, for example, the uses such as mouth-washes as virus infection prevention usefulness of the solution that forms can be dissolved in the water sialic acid oligose-chitosan complex.
And, the invention provides the industrial fiber goods that contain viral trapping agent of the present invention.Fibre product of the present invention, can make filamentary material contain viral sorbent material of the present invention by method physics or chemistry, for example, can be by with sialic acid oligose-chitosan complex of the present invention spraying or invade and soak, rub on weaving cotton cloth etc. into the medium method manufacturing of fiber.Fibre product of the present invention for example can exemplify for filter membrane, mouth mask, towel, handkerchief or as fibre product that processes raw material of its thing etc.Viral sorbent material of the present invention is sneaked into them in various materials of filamentary material etc., can prepare the viral adsorbent product of various goods.
And, chitosan complex of the present invention, mix with water or appropriate solvent or dissolve, as viral sorbent material of the present invention to catch virus with the form of composition, not only can be made into various fibre products such as air cleaner filter membrane, mouth mask, also can by medical facilities, medical facilities articles for use, apparatus, etc. or by coateds such as the livestock of virus pollution, its animal houses, spray or sneak into etc., use as the prevention material of virus infection.
Sialic acid oligose-chitosan complex particularly of the present invention, the composition that constitutes it is all from natural goods, and to nontoxicitys such as people or domestic animals, so viral trapping agent environmental sound of the present invention can be used as washing composition or the viral agent for preventing and eliminating used in home-use or tame animal house, particularly poulty house and uses.
Sialic acid oligose-chitosan complex of the present invention can be easy, when making at an easy rate, also can catch the virus of multiple class effectively.And, contain from the sialic acid oligose-chitosan complex of the present invention of the raw material of natural goods all very safe to environment and human body etc., so have the scope of application widely.
Embodiment
Spendable chitosan comprises the chitosan of peddling as foodstuffs material, reagent etc. among the present invention, or the chitosan for preparing from natural resource with known various methodologies in the past etc. directly uses, and its source is not had special restriction.
Spendable sialyloligosaccharide can exemplify acetylate, hydrosulfate, methide, deoxidant body, dehydrogenation body or the lactosylation thing etc. into sialyl lactose, N-n acetylneuraminic acid n, N-hydroxyacetylneuraminic acid, deamination propylhomoserin, above-mentioned substance among the present invention, comprises the sugar compounds of sialic acid, carboxyl.Can use above-mentioned substance separately, but wish use the mixture of sialyloligosaccharide more than 2 kinds, also wish to use from the breast sialyloligosaccharide of cow's milk, particularly bovine coloctrum or cow's milk for example.And, preferably use the mixture of sialyloligosaccharide and other neutral sugars, for example, from the lactose of breast or colostrum, particularly bovine coloctrum or cow's milk and the mixture of sialyloligosaccharide.At this moment, have only the sialyloligosaccharide of acid sugar to be incorporated into chitosan, after carrying out absorption reaction, can remove neutral sugar by processes such as dialysis, filtrations, can omit the treating process of sialyloligosaccharide from natural goods (for example cow's milk or colostrum), so on operability and manufacturing cost, have favourable one side.
The association reaction of chitosan and sialyloligosaccharide can carry out in the solution of the scope of preferred pH4～7 or the suspension in pH3～10.In detail, make chitosan dissolving or be suspended in the appropriate solvent, add sialyloligosaccharide make its in conjunction with after, through known treating processes such as dialysis, filtration, precipitations, carry out drying under reduced pressure, spraying drying, lyophilize etc., obtain sialic acid oligose-chitosan complex.As the solvent of association reaction, can use acetic acid, citric acid, lactic acid, formic acid, phosphoric acid, hydrochloric acid, nitric acid, sulfuric acid or contain above-mentioned buffered soln, the aqueous solution or organic solution etc. more than a kind or 2 kinds.
Below, further specify the present invention with embodiment.
Embodiment
Embodiment 1
6mg is dissolved in 0.5% acetic acid of 1ml with chitosan (PSH-80, Kiyatsu Aquatic Product Chemical Co., Ltd.).Add dissolved 25mg aqueous solution 0.5ml in advance in sialyl lactose (Sigma company), 25 ℃ were stirred 12 hours.Reaction solution is moved into dialysis tubing (seeing through molecular weight about 14,000), carry out 5 days dialysis (2 exchange/days) with distilled water, the final liquid of lyophilize obtains 11mg lactose-chitosan complex.By
1H-NMR (D
2O, Bruker company, 500MHz) the evaluation structure.
1H-NMR(D
2O、500MHz)δ：
5.121(0.45H，d，J＝3.6Hz，H-1 ofαGlc in SiaLac)，
4.58-4.40(4.5H，m，H-1 of Gal，H-1 ofβGlc in SiaLac，H-1 of GlcN and GlcNAc)，
4.01-3.07(32.9H，m，protons from sugar rings)，
2.80(2.78H，br，H-2 of GlcN)，
2.63(1H，dd，H-3eq of Sia)，
1.97-1.94(4.63H，m，COCH
3)，
1.66(1H，t，H-3ax of Sia)。
Proved from from
1Peak value that near the H-NMR spectrographic 1.66 sialic acid is 3 and integrated value near the peak value of the chitosan glycosamine 2.80, for the glycosamine residue of chitosan, sialyl lactose is adsorbed in 35% ratio on the main chain of chitosan (promptly the mol ratio that exists of glycosamine residue in complex body and sialyl lactose is 1: 0.35).
Embodiment 2
Add the 800ml chloroform the bovine coloctrum 800ml that reclaim in the pasture of prefecture, river clear water raised path between farm fields from Hokkaido and stir degreasing.After repeating 2 recycle-water phase components, add ethanol and be made into 80% ethanolic soln, remove precipitating proteins to reclaim supernatant,, obtain the sugar chain component 16.4g of colostrum further with after its lyophilize with centrifugation.The sugar chain component of gained colostrum is passed through spectrometry mass (MALDI-TOF MS, UltraFLEX, when Brruker Daltonics society) analyzing, identified and be equivalent to lactose (Lac, ((Sia α 2 → 6Gal β 1 → 4Glc) is は 3 ' SL (Sia α 2 → 3Gal β 1 → 4Glc), and the び DSL (spectrum of Sia α 2 → 8Sia α 2 → 3Gal β 1 → 4Glc) again for Sia α 2 → 6Gal β 1 → 4GlcNac), 6 ' SL for Gal β 1 → 4Glc), 6 ' SLN.MALDI-TOF MS[M+Na]
+：
Lac (C12H22011) calculated value: 365.1,
Measured value: 364.8;
6 ' SL or 3 ' SL (C23H39NO19) calculated value: 656.2,
Measured value: 656.0;
6 ' SLN (C25H42N2O19) calculated value: 697.2,
Measured value: 698.0:
DSL (C34H56N2O28 calculated value: 963.8,
Measured value: 963.1.
Next, chitosan (PSH-80, Kiyatsu Aquatic Product Chemical Co., Ltd.) 6mg is dissolved in 0.5% acetic acid of 1ml.Add therein in advance with above-mentioned gained 100mg colostrum dissolved aqueous solution 1ml, 25 ℃ were stirred 48 hours.Reaction solution is moved into dialysis tubing (seeing through molecular weight about 14,000), carry out dialysis (2 exchange/days) on the 5th with distilled water, the final liquid of lyophilize obtains the 9mg sialic acid oligose-chitosan complex.By
1H-NMR (D
2O, Bruker company, 500MHz) the evaluation structure.
1H-NMR (D
2O, Bruker society, 500MHz) δ:
5.12(0.29H，m，H-1 of αGlc in Sialooligosaccharide)，
4.58-4.40(5.6H，m，H-1 of Gal，H-1 of βGlc in Sialooligosaccharide，H-1 of GlcN andGlcNAc)，
4.01-3.07(35.9H，m，protons from sugar rings)，
2.80(4.38H，br，H-2 of GlcN)，
2.63(1H，m，H-3eq of Sia)，
1.97-1.94(5.45H，m，COCH
3)，
1.66(1H，m，H-3ax of Sia)。
Proved from from
1Near the H-NMR spectrographic 1.66 3 peak values of sialic acid and integrated value near the peak value of the chitosan glycosamine 2.80 are adsorbed in 23% ratio for sialyloligosaccharide on the main chain of chitosan (i.e. glycosamine residue in complex body and sialic acid residues have mol ratio be 1: 0.23).
Embodiment 3
Sialic acid oligose-chitosan complex is to the hemagglutination-inhibition test (HI) of avian influenza virus
1. experiment material:
Sialyl lactose-chitosan complex of sample 1: embodiment 1
Sialyloligosaccharide sugar chain-chitosan complex of sample 2: embodiment 2
Sample 3: chitosan (PSH-80, Japanese Kiyatsu Aquatic Product Chemical Co., Ltd.)
H5, H9, three kinds of hypotype antigen hemagglutinating antigens of H7 bird flu
1% chicken red blood corpuscle
96 hole V-type plates
2. experimental technique:
Add the 25ul test sample of the different concns for preparing in advance in the different holes of 96 hole V-type plates, hole respectively, every then hole adds 25ul 4HA antigen respectively, mixing on micro oscillator, and room temperature was placed 60 minutes.After 60 minutes, every hole adds 25ul 1% chicken blood cell again, mixing on micro oscillator, and room temperature was placed 30 minutes.Observe the red cell agglutination situation, no aggegation is judged as (-), the aggegation degree by light to being judged as (+～++ ++) heavily respectively
3. test-results:
Hemagglutination-inhibition test (4 ℃ of conditions)
| The H5 hypotype | Sample concentration (ug/ml) | ||||||
| Sample number into spectrum | 125 | 62.5 | 31.5 | 25 | 20 | 15.75 | 10 |
| 1 | + | - | - | - | - | - | + |
| 2 | ++++ | - | - | - | - | - | + |
| 3 | ++++ | ++++ | ++++ | ++++ | ++++ | ++++ | ++++ |
| The H7 hypotype | |||||||
| Sample number into spectrum | 125 | 62.5 | 31.5 | 25 | 20 | 15.75 | 10 |
| 1 | + | - | - | - | - | + | + |
| 2 | ++ | ++ | - | - | - | + | + |
| 3 | ++++ | ++++ | ++++ | ++++ | ++++ | ++++ | ++++ |
| The H9 hypotype | |||||||
| Sample number into spectrum | 125 | 62.5 | 31.5 | 25 | 20 | 15.75 | 10 |
| 1 | + | + | - | - | - | - | + |
| 2 | ++ | - | - | - | - | + | + |
| 3 | ++++ | ++++ | ++++ | ++++ | ++++ | ++++ | ++++ |
The result shows that sialic acid oligose-chitosan complex has tangible blood clotting restraining effect to avian influenza virus
The present invention is with easy method and use raw material from the natural goods of environmental sound, sialic acid oligose-chitosan complex is provided, sialic acid oligose-chitosan complex of the present invention, various viruses are had antiviral activity, virus seizure activity, and effective as the material of viral passivation and seizure.
The sialic acid oligose-chitosan complex of invention, because of to human body and animal safety, environmental sound, and cost is low, virus is had strong activity, thus can be used as be used in the type material that pollution that defence, prevention, diagnosis cause by various viruses and disease use industrial.
Claims (9)
1. sialic acid oligose-chitosan complex is characterized in that: sialyloligosaccharide is incorporated into chitosan with the form of ionic bond and forms complex body.
2. sialic acid oligose-chitosan complex according to claim 1, it is characterized in that: described sialyloligosaccharide is one or more of following material: sialic acid, sialic acid derivative, contain sialic sugar chain, the sugar chain that contains sialic acid derivative, the compound that contains as functional group and intramolecularly with sialic acid or sialic acid derivative.
3. sialic acid oligose-chitosan complex according to claim 1 is characterized in that: described sialyloligosaccharide derives from the colostrum of mammal.
4. sialic acid oligose-chitosan complex according to claim 1 is characterized in that: described sialyloligosaccharide is for being the sialyloligosaccharide of representative with the sialyl lactose.
5. as the preparation method of one of claim 1-4 described sialic acid oligose-chitosan complex, it is characterized in that: utilize the cationic of the anionic property and the chitosan of sialyloligosaccharide, make sialyloligosaccharide and chitosan carry out ionic bond.
6. the preparation method of sialic acid oligose-chitosan complex according to claim 5 is characterized in that: described sialyloligosaccharide is for from the isolating mixture as enriched material of natural goods.
7. the preparation method of sialic acid oligose-chitosan complex according to claim 5, it is characterized in that: described mixture is that the sialyl lactose that obtains in the cow's milk is the mixture of the sialyloligosaccharide of representative.
8. viral sorbent material is characterized in that, comprises the described sialic acid oligose-chitosan complex as one of claim 1-4.
9. the medicine of antiviral usefulness, sanitary product or mechanicals is characterized in that, are effective constituent with the described viral sorbent material of claim 8.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101686716B (en) * | 2007-06-25 | 2013-12-18 | 帝斯曼知识产权资产管理有限公司 | Prebiotics |
| CN103551562A (en) * | 2013-10-21 | 2014-02-05 | 中国科学院微生物研究所 | Sialyloligosaccharide-gold nano particle and preparation method and applications thereof |
| CN103834033A (en) * | 2013-12-04 | 2014-06-04 | 江南大学 | Preparation method of polysialic acid-chitosan derivative hydrogel |
| CN104406954A (en) * | 2014-11-26 | 2015-03-11 | 中国科学院微生物研究所 | Surface enhanced Raman spectra substrate for detecting avian influenza virus, and application thereof |
| CN114397448A (en) * | 2021-04-01 | 2022-04-26 | 苏州育德扬生物技术有限公司 | Preparation of sugar functionalized nanoparticles and application of sugar functionalized nanoparticles in influenza virus detection |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101686716B (en) * | 2007-06-25 | 2013-12-18 | 帝斯曼知识产权资产管理有限公司 | Prebiotics |
| CN103551562A (en) * | 2013-10-21 | 2014-02-05 | 中国科学院微生物研究所 | Sialyloligosaccharide-gold nano particle and preparation method and applications thereof |
| CN103551562B (en) * | 2013-10-21 | 2015-07-15 | 中国科学院微生物研究所 | Sialyloligosaccharide-gold nano particle and preparation method and applications thereof |
| CN103834033A (en) * | 2013-12-04 | 2014-06-04 | 江南大学 | Preparation method of polysialic acid-chitosan derivative hydrogel |
| CN103834033B (en) * | 2013-12-04 | 2016-03-23 | 江南大学 | The preparation method of a kind of Polysialic acid-chitosan derivatives hydrogel |
| CN104406954A (en) * | 2014-11-26 | 2015-03-11 | 中国科学院微生物研究所 | Surface enhanced Raman spectra substrate for detecting avian influenza virus, and application thereof |
| CN104406954B (en) * | 2014-11-26 | 2017-06-30 | 中国科学院微生物研究所 | A kind of Raman spectrum base thing for detecting avian influenza virus and its application |
| CN114397448A (en) * | 2021-04-01 | 2022-04-26 | 苏州育德扬生物技术有限公司 | Preparation of sugar functionalized nanoparticles and application of sugar functionalized nanoparticles in influenza virus detection |
| CN114397448B (en) * | 2021-04-01 | 2024-01-19 | 苏州育德扬生物技术有限公司 | Preparation of sugar functionalized nano particles and application of sugar functionalized nano particles in influenza virus detection |
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| CN100494368C (en) | 2009-06-03 |
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