CN107254016A - A kind of preparation method of difunctional compolymer/nano grain inhibiting agent and the suppression and the application of removing toxic substances assembled in amyloid beta protein - Google Patents
A kind of preparation method of difunctional compolymer/nano grain inhibiting agent and the suppression and the application of removing toxic substances assembled in amyloid beta protein Download PDFInfo
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Abstract
The present invention one is related to kind of the preparation method of difunctional compolymer/nano grain inhibiting agent and the suppression and the application of removing toxic substances assembled in amyloid beta protein；The dual-functional nanometer grain inhibiting agent with metal chelation abilities and suppression albumen ability of aggregation is synthesized by free radicals copolymerization reaction using the connection product GMA IDA of iminodiacetic acid (IDA) and GMA (GMA) as monomers such as monomer and N N-isopropylacrylamides (NIPAm), N-tert-butyl acrylamides (TBAm).The average grain diameter of the difunctional compolymer/nano particle of this method synthesis is 80 120nm.The metal ion-chelant capacity of the nano particle of this method synthesis is more than 700 μm of ol/g, can chelate the metal ion in environment, is a kind of new drug raw material of potential treatment alzheimer's disease while suppressing A β 42 fibrosis accumulation process.
Description
Technical field
The present invention relates to a kind of compolymer/nano particle suppression with metal-chelating and the dual-use function for suppressing fibrosis aggregation
The preparation method of preparation and the application in terms of the suppression and removing toxic substances that metal inducement amyloid beta protein is assembled.
Background technology
Alzheimer's disease is most commonly seen senile dementia, and its illness shows as cognition dysfunction, memory capability
The symptoms such as decline, emotional instability.Numerous studies show in AD pathogenic processes along with amyloid beta (amyloid β-
Protein, A β) accumulation process, fibrous patch is produced in patient's intracerebral neuronal cell.Therefore, it is considered that A β gather
There are close ties between integrated fibers and AD morbidities.
A β be after cross-film amyloid precusor protein (APP) elder generation through β-and gamma-secretase shearing obtain.But due to γ-point
The restriction enzyme site for secreting enzyme is different, therefore can produce the A beta polypeptides with different C ' end amino acids, such as A β 43, A β
42、Aβ40、Aβ38、Aβ37.Wherein, content highests of the A β 40 in human brain, account for A beta polypeptides sum 50%-70%, and its
The A β contents of its species are all very low.And found in pathological research, A β 42 are the chief components of amyloid plaques, are had
The stronger ability for being gathered into fiber, and it was found that it secretes more in familial AD patient.Therefore, A β 40 and A β 42 are existing
Albumen is predominantly targeting in research AD treatment methods.
Zinc ion is in close relations with nervous system, and brain is one of internal zinc content highest organ.Research shows, in
About 85% zinc is mainly combined with metalloprotein in pivot nervous system, and Free Zinc contains in big intracerebral average content for Zn in serum
10 times of amount.Appropriate concentration zinc ion is to maintaining internal eubolism to play a key effect.Have been reported that at present and show AD patient's brain
The zinc ion level of interior hippocampus and amygdaloid nucleus is about higher by 30% than normal person, and this is probably due to heavy metal ion such as zinc ions
Inducing action to protein aggregation is relevant.Zinc ion and A β quickly combine to form the zinc-A beta peptide aggregations body that is unable to fibrosis or
Zinc ion is bonded directly to fiber ends, so as to suppress A betas.Generally believe that the aggregation of this zinc ion induction is made at present
There is stronger cytotoxicity with the aggregation formed.
To overcome metal inducement A beta peptide aggregations, the method that a kind of " metal-chelating treatment " is proposed in recent years, this method is intended to
Metal-chelator is added, so as to eliminate the A beta peptide aggregations of metal ion induction.Some metal-chelators such as ethylenediamine tetra-acetic acid two
Sodium (EDTA), enteroquinol (CQ), CQ structural derivatives PBT-2, A β 42 His-Ala mutant etc. can be with
Effectively suppress the aggregation of the aβ protein of metal ion induction.But these traditional small molecule metal-chelators have it is non-specific
Property, toxic side effect and relatively low blood-brain barrier penetrability.
Iminodiacetic acid (IDA) is a kind of chemical raw material, is normally used for the manufacture of the agricultural chemicals such as glyphosate.Given birth to soda acid
Into salt, go back and various metals formation chelate.Using IDA metal-chelating performance, had been reported in chromatographic isolation field by it
Applied to the raw material of ion exchange resin, and do not had been reported that in biomedicine field.
In addition, current " metal-chelating treatment " method primarily directed to the ion domain of the existence in high metal concentration to gold
The albumen aggregation of category induction is suppressed, so that delay the metal inducement of aβ protein to assemble, but to A β formation amyloid fibers
And its suppression of caused toxicity and detoxifying effect it is unsatisfactory.The problem of for A β formation amyloid fibers, Hong-chen
Liu etc. has synthesized a series of compolymer/nano particles with negative electrical charge as formed by including acrylic monomers copolymerization, and obtains
Nano grain surface negative electrical charge assembles the inhibiting mechanism for producing inhibitory action to A beta-amyloyds, and has obtained having and suppress to conciliate
Nano particle inhibitor (the Negatively charged hydrophobic nanoparticles inhibit of toxic action
amyloid β-protein fibrillation:The presence of an optimal charge density[J]
.Reactive and Functional Polymers,2016,103:108-116.).To further expand compolymer/nano particle
Application in terms of the A beta peptide aggregations in the presence of metal, the present invention synthesizes GMA-IDA monomers using iminodiacetic acid (IDA)
(Synthesis of Magnetic Chelator for High-Capacity Immobilized Metal Affinity
Adsorption of Protein by Cerium Initiated Graft Polymerization,Langmuir,2005,
21,6987-6994) compolymer/nano particle, is modified using it, with metal-chelating and the dual work(of suppression fibrosis aggregation
Energy.Nano particle inhibitor preparation technology is simple, and good biocompatibility is highly suitable for what is assembled as amyloid beta protein
The application of medicine and health products.
The content of the invention
It is an object of the invention to overcome the deficiencies in the prior art, further exploitation has metal chelating groups and negative electrical charge
The dual-functional nanometer grain inhibiting agent of surface modification.There is the nano particle metal-chelating to act on and suppress albumen aggregation simultaneously
Effect, can effectively suppress under metal ion existence condition aβ protein aggregation.Simultaneously amyloid is being used as there is provided the present invention
The medicine of albumen aggregation and the purposes of health products.
Technical scheme is summarized as follows：
Connection product GMA-IDA of the invention by iminodiacetic acid (IDA) and GMA (GMA)
Pass through as monomers such as monomer and NIPA (NIPAm), N-tert-butyl acrylamides (TBAm) free-radical polymerized anti-
It should synthesize with metal chelation abilities and suppress the dual-functional nanometer grain inhibiting agent of albumen ability of aggregation.What this method was synthesized receives
The average grain diameter of rice grain is 80-120nm.The metal ion-chelant capacity of the nano particle of this method synthesis reaches 700-800 μ
Mol/g, can chelate the metal ion in environment, while suppressing A β 42 fibrosis accumulation process.
Above-mentioned GMA-IDA structural formula is as follows：
The structural formula of above-mentioned dual-functional nanometer particle is as follows：
Nano particle average grain diameter 80-120nm, nanoparticle metal ion chelating capacity reaches 700-800 μm of ol/g, its
The ratio of x, y, z is 38-53 in middle chemical formula：35-50：5-20.
The preparation method of the difunctional compolymer/nano particle of the present invention, its step is as follows：
1) the connection product GMA-IDA of synthesizing methyl glycidyl acrylate and iminodiacetic acid；
2) by polymerized monomer N ,-N '-methylene bisacrylamide, N-tert-butyl acrylamide, NIPA according to
1-5：38-53：35-50 ratio and GMA-IDA mixing, wherein the mol ratio for controlling GMA-IDA monomers to account for total monomer is 5-
20%.
3) to above-mentioned steps 2) in mixed liquor in add surfactant, such as sodium hexadecyl sulfate, and make surface
The final concentration of 0.2-0.4mg/mL of activating agent；
4) by above-mentioned steps 3) mixed liquor it is ultrasonically treated be allowed to be completely dissolved, and it is mixed to discharge to be passed through nitrogen 10-60min
Close the air in liquid；
5) to above-mentioned steps 4) in mixed liquor in add the starting copolyreaction of initiator sodium peroxydisulfate.Under nitrogen protection
Difunctional compolymer/nano particle is obtained after reacting 10-15h in water bath with thermostatic control shaking table.
The particle diameter for the difunctional compolymer/nano particle that the present invention is obtained is 80-120nm, the surface δ current potentials in water are about-
40mV, negative charged surface result in the intermolecular electrostatic repulsion of nano particle and the albumen of A β 42, so as to more effectively press down
A β processed fibrosis aggregation；In addition, the substantial amounts of metal chelating groups of nanometer dielectric surface, its metal ion-chelant capacity>700μ
Mol/g, can effectively adsorb the metal ion in environment, hinder the amyloid aggregation of metal ion induction；Suppress A β simultaneously
42 fibrosis accumulation process, is a kind of new drug raw material of potential treatment alzheimer's disease.
Difunctional compolymer/nano particle of the present invention is used for the suppression of metal ion induction amyloid aggregation
Method has the following advantages that：
First, compared with common metal chelating agent, difunctional compolymer/nano particle of the invention has more preferable biofacies
Capacitive；
Second, compared with previous single function nano grain inhibiting agent, difunctional compolymer/nano particle of the invention has
Good metal chelation abilities, " the metal-chelating treatment " assembled available for amyloid.
3rd, the A β 42 that difunctional compolymer/nano grain inhibiting agent of the invention can suppress metal ion induction simultaneously gather
Collection and the aggregation of the fibrosis of A β 42.
4th, difunctional compolymer/nano grain inhibiting agent of the present invention can be reduced produced by the accumulation process of A β 42
Cytotoxic effect.
Brief description of the drawings
Fig. 1：With zinc ion and zinc ion and difunctional compolymer/nano grain inhibiting agent mixture co-incubation in embodiment 4
The aggregations of A β 42 ThT fluorescence intensities.
Fig. 2：With zinc ion and zinc ion and difunctional compolymer/nano grain inhibiting agent mixture co-incubation in embodiment 5
The aggregations of A β 42 transmission electron microscope picture.
Fig. 3：With zinc ion and zinc ion and difunctional compolymer/nano grain inhibiting agent mixture co-incubation in embodiment 6
The aggregations of A β 42 circular dichroism spectrogram.
Fig. 4：With zinc ion and zinc ion and difunctional compolymer/nano grain inhibiting agent mixture co-incubation in embodiment 7
A β 42 SH-SY5Y cell survival rate is influenceed to scheme.
Embodiment
Following example will be further described to the method that the present invention is provided：
Embodiment 1：The synthesis of difunctional compolymer/nano grain inhibiting agent.
Copolymer nano particle is prepared by free radicals copolymerization reaction.(1) by N ,-N '-methylene bisacrylamide, the tert-butyl group
Acrylamide, NIPA and GMA-IDA 1%, 53%, 41%, 5% mixing in molar ratio respectively.In mixture,
Surfactant sodium hexadecyl sulfate 15mg is added, mixed liquor final volume is 50mL.Obtained solution blows about in nitrogen
30min, with emptying air.60mg/mL sodium peroxydisulfate initiators are added, under nitrogen protection in 25 DEG C of 160rpm shaking tables reactions
12h.Resulting polymers dialyse 14 days (changing water at least twice daily) with MilliQ water to remove unreacted monomer after reaction.Receive
Rice grain is lyophilized, and to be stored in 4 DEG C of refrigerators stand-by.The hydraulics particle diameter of nano particle made from the condition is 118 ± 6nm, surface
Zeta potential is -37 ± 5mV.
Embodiment 2：The synthesis of difunctional compolymer/nano grain inhibiting agent.
Copolymer nano particle is prepared by free radicals copolymerization reaction.(1) by N ,-N '-methylene bisacrylamide, the tert-butyl group
Acrylamide, NIPA and GMA-IDA 2%, 40%, 48%, 10% mixing in molar ratio respectively.Mixture
In, surfactant sodium hexadecyl sulfate 10mg is added, mixed liquor final volume is 50mL.Obtained solution blows in nitrogen
About 60min, with emptying air.60mg/mL sodium peroxydisulfate initiators are added, under nitrogen protection in 25 DEG C of 160rpm shaking tables reactions
15h.Resulting polymers dialyse 14 days (changing water at least twice daily) with MilliQ water to remove unreacted monomer after reaction.Receive
Rice grain is lyophilized, and to be stored in 4 DEG C of refrigerators stand-by.The hydraulics particle diameter of nano particle made from the condition is 88 ± 6nm, surface
Zeta potential is -41 ± 8mV.
Embodiment 3：The synthesis of difunctional compolymer/nano grain inhibiting agent.
Copolymer nano particle is prepared by free radicals copolymerization reaction.(1) by N ,-N '-methylene bisacrylamide, the tert-butyl group
Acrylamide, NIPA and GMA-IDA 5%, 38%, 37%, 20% mixing in molar ratio respectively.Mixture
In, surfactant sodium hexadecyl sulfate 20mg is added, mixed liquor final volume is 50mL.Obtained solution blows in nitrogen
About 10min, with emptying air.60mg/mL sodium peroxydisulfate initiators are added, under nitrogen protection in 25 DEG C of 160rpm shaking tables reactions
10h.Resulting polymers dialyse 14 days (changing water at least twice daily) with MilliQ water to remove unreacted monomer after reaction.Receive
Rice grain is lyophilized, and to be stored in 4 DEG C of refrigerators stand-by.The hydraulics particle diameter of nano particle made from the condition is 80.0 ± 5nm, surface
Zeta potential is -49 ± 7mV.
Embodiment 4：With zinc ion and zinc ion and the A β of difunctional compolymer/nano grain inhibiting agent mixture co-incubation
The ThT fluorescence intensities of 42 aggregations.
First, by the solid powders of A β 42 so that in the dissolving of 1mg/mL concentration and HFIP, ultrasonic disperse 20min is gathered with removing A β 42
Collective, then standing 2h makes the fully dissolvings of A β 42.Then by A β 42 HFIP solution in 4 DEG C, refrigerated centrifuge under 12000g
20min, takes 75% supernatant to be freeze-dried.The solid powders of A β 42 after will be lyophilized store stand-by in -20 DEG C of low temperature refrigerators.
Before use, the solid powders of A β 42 are redissolved in 20mM NaOH, with after 4 DEG C, refrigerated centrifuge 20min under 12000g,
75% supernatant is taken to be freeze-dried.It is subsequently added not metal ion Hepes buffer As (20mM, pH 7.4), or containing metal
IONS OF H epes buffer Bs (7.4,25 μM of ZnCl of 20mM, pH), or add 50 μ g/mL single function nano particle and (pass through
Document Negatively charged hydrophobic nanoparticles inhibit amyloid β-protein
fibrillation:The presence of an optimal charge density[J].Reactive and
Functional Polymers,2016,103:Method is synthesized in 108-116.) Hepes buffer Bs in, or add 50 μ
In the Hepes buffer Bs of difunctional compolymer/nano particle prepared by the g/mL present invention, final A β concentration is 25 μM.At 37 DEG C,
48h is cultivated in 160rpm shaking tables.200 μ L samples are added in 40 μM of ThT solution of 2mL, Fluorescence spectrophotometer is used
(Perking Elmer LS-55, MA, USA) is measured to the albumen of A β 42 aggregation situation, is determined and is carried out under the conditions of 25 DEG C,
Wherein exciting light is 440nm, and transmitting light is 480nm, slit width 5nm.Experimental result takes the average value of three measure.
By Fig. 1 it can be seen that, the ThT fluorescence intensities that A β 42 containing zinc ion assemble substantially are reduced, and add zinc ion
Significantly rise with the A β 42 of the sample of the difunctional compolymer/nano particle ThT fluorescence assembled, but and add zinc ion and single function
The ThT fluorescence that the A β 42 of the sample of nano particle assemble rises unobvious.This explanation zinc ion induction A β 42, which assemble to be formed, not to be had
There is the aggregation of β lamellar structures；The aggregations of A β 42 that single function nano particle can not suppress zinc ion induction are formed；And it is difunctional
Compolymer/nano particle can be with chelated zinc ions, and the aggregations of A β 42 for suppressing zinc ion induction are formed.
Embodiment 5：With zinc ion and zinc ion and the A β of difunctional compolymer/nano grain inhibiting agent mixture co-incubation
The varying topography of 42 aggregations.
Handled according to method same as Example 4 and cultivate the samples of A β 42.After 48h has been cultivated, 100 μ L A β are taken to train
5-10 μ L solution is added dropwise on the carbon film copper mesh of 400 mesh nutrient solution, ultrasonic 10min, spontaneously dries.Treat that the solution on copper mesh will
When dry not dry, surplus liquid is sopped up after Salkowski's solution (pH 6.5) the negative staining 30s for being added dropwise 2%, is spontaneously dried.Obtained sample
Observed using JEM-100CXII transmission electron microscopes (accelerating potential 100kV).Choose the image that scale after amplification is 100nm
Observed, as a result as shown in Figure 2.
By Fig. 2 it can be seen that, add zinc ion A beta peptide aggregations into random metal-protein masses, add zinc
The morphology of the aggregate of A β 42 of ion and single function nano particle substantially with add the A beta peptide aggregations of zinc ion into random metal-egg
White aggregation is consistent, and the A β 42 that this instruction sheet function nano particle can not suppress metal ion induction assemble.And add zinc from
The aggregations of A β 42 of sub and difunctional compolymer/nano particle form short and thick fibrilla aggregates.This illustrates IDA not only
The A β 42 for preventing metal ion to induce assemble, and inhibit A β fibrosis to assemble to a certain extent.
Embodiment 6：With zinc ion and zinc ion and the A β of difunctional compolymer/nano grain inhibiting agent mixture co-incubation
The secondary structure change of 42 aggregations
According to the processing of the identical method of example 4 and cultivating the albumen of A β 42.It is 0.1mm to take 300 μ L nutrient solutions to add light path
Silica dish in, under extreme ultraviolet wavelength (190-260nm) observe.Wherein sweep bandwidth is 1nm, and sweep speed is 100nm/
min.Continuously scan three times and take averagely as measurement result, measurement result is as shown in Figure 3.
From figure 3, it can be seen that after 48h is cultivated, sample containing only A β 42 200nm or so there is positive maximum and
219nm or so goes out negative minimum.The curve shows to form β-lamellar structure in sample.And with 25 μM of zinc ion co-incubations
There is negative minimum in 200nm or so and has a very weak negative value in 220nm or so in sample, and this shows that sample is induced in zinc ion
Under form irregular secondary structure.In the sample for adding single function nano particle, collection of illustrative plates and the result one for adding zinc ion
Cause, this shows that single function nano particle can not suppress the formation of the irregular secondary structures of A β 42 of metal ion induction.Adding
In the sample of difunctional compolymer/nano particle, curve has minimum value at 215-218nm, and this is probably α-helix to β-piece
The second structure characteristic of layer conversion.This shows that difunctional compolymer/nano particle can prevent the formation zinc ion induction of the albumen of A β 42
Irregular secondary structure, and disturb the formation of β-lamellar structure.
Embodiment 7：The A β 42 of zinc ion and zinc ion and difunctional compolymer/nano grain inhibiting agent mixture co-incubation
Influence to SH-SY5Y cell survival rate.
Employ SH-SY5Y cell lines (DMEM/F12 culture mediums, wherein 20%FBS, 2mmol/L glutamine, 100U/
ML is dual anti-).Cell is cultivated in 37 DEG C in the constant incubator containing 5%CO2.
Cytoactive is analyzed using MTT detections.Specific experiment step is as follows：Cell culture fluid is added into 96 orifice plates
In, wherein adding 90 μ L cell liquid per hole, the cell density per hole is 5000.Cultivated in incubator after 24h, add 10 μ L
250 μM of zinc ion mother liquor, or single function nano particle containing 250 μM of zinc ions and 500 μ g/mL (pass through document
Negatively charged hydrophobic nanoparticles inhibit amyloid β-protein
fibrillation:The presence of an optimal charge density[J].Reactive and
Functional Polymers,2016,103:Method is synthesized in 108-116.) mother liquor, or contain 250 μM of zinc ions and 500 μ
Difunctional compolymer/nano particle mother liquor prepared by the g/mL present invention.Final zinc ion concentration is 25 μM, and nanoparticle concentration is
50μg/mL.Continue to remove culture medium after cultivating 48h, cleaned, then added in every hole using Hepes buffer solutions (pH 7.4)
100 μ L serum free mediums and 10 μ L MTT solution (6mg/mL), continuation cultivate 4h in 37 DEG C of incubators.
Before detection, culture medium is centrifuged off first by flat board, is subsequently added 100 μ L DMSO dissolvings and is attached to orifice plate
The cell at bottom.96 orifice plate lucifuges are centrifuged into 10min, after after purple crystals all dissolving, it is determined 570nm's using ELIASA
Absorbance.Cytoactive is calculated by absorbance.Wherein not celliferous sample is blank group (0%), without A β 42 and suppression
Agent, the only sample containing cell are control group (100%).Each sample sets 6 multiple holes to determine, and measurement result is averaged.
As a result it is as shown in Figure 4.
When without zinc ion, A β 42 addition can cause cytoactive to be reduced to 64.7%, and the A β 42 containing zinc ion
The cytoactive of sample is reduced to 47.6%, hence it is evident that less than the cytoactive of the samples of A β 42 without zinc ion.This show zinc from
The albumen aggregation of son induction has higher cytotoxicity.Adding the sample of difunctional compolymer/nano particle has higher cell
Active (86.3%), not only far above the sample of single function nano particle, and cytoactive is apparently higher than the sample for only adding A β 42
Product.This shows that difunctional compolymer/nano particle has not only chelated zinc ion, eliminates the toxicity of metal inducement protein aggregation, together
When A β 42 fibrosis is assembled caused by cytotoxicity also there is good detoxication.
The present invention is proposed using radical copolymerization method, utilizes metal-chelating function monomer, hydrophobic function monomer
Etc. having synthesized the difunctional compolymer/nano particle with metal chelating groups, and difunctional compolymer/nano particle is proposed in suppression
The application of metal inducement beta-amyloid aggregation and its relevant cytotoxic, and in alzheimer disease drug and health products
Application in terms of exploitation.It is described by live preferred embodiment, person skilled can substantially not depart from this
Method described herein is modified in the content of the invention, spirit and scope or suitably change is with combining, to realize skill of the present invention
Art.In particular, all similar replacements and change are apparent to those skilled in the art, he
Be considered as being included in spirit of the invention, scope and content.
Claims (4)
1. a kind of difunctional compolymer/nano grain inhibiting agent；It is characterized in that its structural formula is：
Synthesis difunctional compolymer/nano particle average grain diameter be 80-120nm, this method synthesis nano particle metal from
Sub- chelating capacity is more than 700 μm of ol/g, and the ratio of x, y, z is 38-53 wherein in chemical formula：35-50：5-20.
2. the preparation method of the bifunctional copolymer nano particle inhibitor described in claim 1, it is characterized in that preparation process is such as
Under：
1) the connection product GMA-IDA of synthesizing methyl glycidyl acrylate and iminodiacetic acid；
2) by polymerized monomer N ,-N '-methylene bisacrylamide, N-tert-butyl acrylamide, NIPA is according to 1-5：
38-53：35-50 ratio and GMA-IDA mixing, wherein the mol ratio for controlling GMA-IDA monomers to account for total monomer is 5-20%.
3) to above-mentioned steps 2) in mixed liquor in add surfactant, such as sodium hexadecyl sulfate, and make surface-active
The final concentration of 0.2-0.4mg/mL of agent；
4) by above-mentioned steps 3) mixed liquor it is ultrasonically treated be allowed to be completely dissolved, and be passed through nitrogen 10-60min to discharge mixed liquor
In air；
5) to above-mentioned steps 4) in mixed liquor in add initiator starting copolyreaction.Shaken under nitrogen protection in water bath with thermostatic control
In bed difunctional compolymer/nano particle is obtained after reaction 10-15h.
3. the difunctional compolymer/nano particle described in claim 1 is suppressing metal inducement beta-amyloid aggregation and its correlation
The application of cytotoxicity.
4. the difunctional compolymer/nano particle described in claim 1 is in terms of for alzheimer disease drug and health products exploitation
Application.
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| CN108905639A (en) * | 2018-07-12 | 2018-11-30 | 中北大学 | A kind of function polysulfone millipore filtering membrane preparation method of effectively catching heavy metal ion |
| CN109954128A (en) * | 2019-03-08 | 2019-07-02 | 南开大学 | It is a kind of to may interfere with beta-amyloid aggregation and assist the functionalized nano gel removed and preparation method |
| CN110302188A (en) * | 2019-05-10 | 2019-10-08 | 郑州大学 | A kind of preparation method of the pharmaceutical composition of the hydrophobic associated polymer of anti-Alzheimer disease |
| CN110302188B (en) * | 2019-05-10 | 2021-12-10 | 郑州大学 | Preparation method of hydrophobic association polymer drug composition for resisting Alzheimer's disease |
| CN111122529A (en) * | 2019-12-30 | 2020-05-08 | 陕西师范大学 | Application of gold nanoclusters in detection of protein amyloid fibrosis in biological sample and screening of inhibitor |
| CN111122529B (en) * | 2019-12-30 | 2022-07-08 | 陕西师范大学 | Application of gold nanoclusters in detection of protein amyloid fibrosis in biological sample and screening of inhibitor |
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