CN108514564A - Abiduoer functionalized nano selenium with anti-H1N1 influenza activities and its preparation and application - Google Patents
Abiduoer functionalized nano selenium with anti-H1N1 influenza activities and its preparation and application Download PDFInfo
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- CN108514564A CN108514564A CN201810382111.1A CN201810382111A CN108514564A CN 108514564 A CN108514564 A CN 108514564A CN 201810382111 A CN201810382111 A CN 201810382111A CN 108514564 A CN108514564 A CN 108514564A
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
- A61K31/404—Indoles, e.g. pindolol
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- A—HUMAN NECESSITIES
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- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
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- B—PERFORMING OPERATIONS; TRANSPORTING
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- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
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Abstract
The present invention discloses a kind of Abiduoer functionalized nano selenium with anti-H1N1 influenza activities and its preparation and application, belongs to nanocomposite technical field.The Abiduoer functionalized nano selenium is realized using nanometer selenium load Abiduoer, is included the following steps：Abiduoer solution is added into nanometer selenium solution, dialyses, wash after being stirred to react, is dry, obtains nanometer selenium delivery Abiduoer nanometer medicine-carried system, i.e. Abiduoer functionalized nano selenium.Nanometer selenium has good antivirus action as pharmaceutical carrier, and Germicidal efficacy nanometer selenium load Abiduoer, which can cooperate with, inhibits H1N1 influenza viruses to replicate.Nanometer selenium loads Abiduoer and H1N1 infection cells is inhibited to have important academic significance as the treatment of antiviral drugs enhanced sensitivity, and reference is provided for the research of open clinical antiviral drug Mechanism.
Description
Technical field
The invention belongs to nanocomposite technical field, more particularly to it is a kind of with anti-H1N1 influenza activities Ah
Than Duo Er functionalized nanos selenium and its preparation and application.
Background technology
The propagation of H1N1 influenza viruses is infected brings serious hidden danger and disaster to human society.It is inoculated with influenza virus vaccine
People at highest risk's flu episode rate and the death rate can be reduced, but currently available vaccines, existing vaccines lacks specificity to the prevention of new virus strain,
Cause prevention or the treatment of clinically influenza virus bad, after influenza virus mutant, new subtype causes to infect, and is not easy effectively to control
System.Specific influenza viral vaccine cannot meet global demand amount during flu outbreak, be controlled with influenza virus Antiviral Effect
It treats particularly important.
Abiduoer is a kind of weakly basic drugs, it can improve the pH of intracellular in host cell, prevent virus from complete
The process of film forming fusion releasing virus genome, effectively inhibits virus infection.The essential trace elements of the human body selenium can pass through dimension
Hold the occurrence and development process that the number of ways such as immunity of organisms and redox equilibrium participate in viral disease.Host cell
Interior Se content can influence the mutation, duplication and virulence of many intrusive viruses.Viral drug resistance treatment zone is given in the appearance of nanotechnology
Carry out dawn.
Invention content
In order to overcome the disadvantages and deficiencies of the prior art, the purpose of the present invention is to provide one kind having anti-H1N1 influenzas disease
The preparation method of the Abiduoer functionalized nano selenium of cytotoxic activity.
Another object of the present invention is to provide the Abiduoer functionalized nano selenium obtained by above-mentioned preparation method.
It is still another object of the present invention to provide the applications of above-mentioned Abiduoer functionalized nano selenium.
Nanometer selenium has the characteristics that high bioactivity, hypotoxicity as pharmaceutical carrier.The present invention using nanometer selenium load Ah
Being cooperateed with than Duo Er inhibits H1N1 influenza viruses to replicate.For clinical antiviral enhanced sensitivity treatment provide new route of administration and science according to
According to.
The purpose of the invention is achieved by the following technical solution：
The present invention provides a kind of preparation method of the Abiduoer functionalized nano selenium with anti-H1N1 influenza activities,
It is to be realized using nanometer selenium load Abiduoer, includes the following steps：
Abiduoer solution is added into nanometer selenium solution, dialyses, wash after being stirred to react, is dry, obtaining nanometer selenium
Deliver Abiduoer nanometer medicine-carried system, i.e. Abiduoer functionalized nano selenium.
Preferably, in reaction system, the mass ratio of Abiduoer and nanometer selenium is 1: (2~8)；More preferably 1: 4.
In reaction system, final concentration of 10~40 μ g/mL of Abiduoer；Preferably 20 μ g/mL.
Specifically comprise the following steps：
(1) 2mL 50mM vitamin c solutions is taken to be added dropwise to a concentration of 0.1M sodium selenites (Na of 0.25mL2SeO3) molten
In liquid, deionized water, which is added, makes final volume be 25mL, and magnetic agitation 2 hours obtains nanometer selenium solution after purification after dialysis；
(2) the nanometer selenium solution for taking step (1) to prepare, is added the Abiduoer solution of a concentration of 20mg/mL, makes reactant
The final concentration of Abiduoer reaches 10~40 μ g/mL in system；Magnetic agitation, the solution after reaction is by dialysis, centrifugation, deionization
Washing three times, sample are freeze-dried to obtain nanometer selenium delivery Abiduoer nanometer medicine-carried system, i.e. Abiduoer functionalized nano
Selenium.
Preferably, the time of the magnetic agitation described in step (2) is 1h.
A kind of Abiduoer functionalized nano selenium with anti-H1N1 influenza activities, is prepared by above-mentioned preparation method
It obtains.
The Abiduoer functionalized nano selenium with anti-H1N1 influenza activities is preparing anti-H1N1 influenzas disease
Application in cytotoxic activity product.
The effective concentration of the Abiduoer functionalized nano selenium is 80~100 μ g/mL；Preferably 100 μ g/mL.
The present invention mechanism be：
Deliver Abiduoer nanometer medicine-carried system by obtaining the nanometer selenium that disperses evenly and stably, nano-scale selenium deliver Ah
Inhibit virus infected cell effect than Duo Er.There is nanometer selenium good antiviral activity, nanometer selenium to load Abiduoer and inhibit
H1N1 infection cells have important academic significance as the treatment of antiviral drugs enhanced sensitivity, are open clinical antiviral drug Mechanism
Research provides reference.
The present invention has the following advantages and effects with respect to the prior art：
Nanometer selenium has good antivirus action as pharmaceutical carrier, and Germicidal efficacy nanometer selenium loads Abiduoer can
Collaboration inhibits H1N1 influenza viruses to replicate.
Description of the drawings
Fig. 1 is preparation and the characterization of nanometer selenium delivery Abiduoer；Wherein, figure A and B figures are the dindar effects of Se@ARB
It answers；A and b is the transmission electron microscope picture of SeNPs and Se@ARB respectively in figure C；Scheme D be elemental analysis, wherein Se elements account for 72%, C,
N and O is respectively 9%, 4% and 5% from ARB.
Fig. 2 is that SeNPs delivery Abiduoers detect (MTT) to mdck cell survival rate；Wherein, Control：Normal cell
Control group, Virus：Virus infected cell group, Virus+ARB：Virus infected cell+Abiduoer group, Virus+SeNPs：Disease
Malicious infection cell+nanometer selenium group, Virus+Se@ARB：Virus infected cell+nanometer selenium delivers Abiduoer group.
Specific implementation mode
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited
In this.
H1N1 influenza viruses used in embodiment are purchased from Wuhan Virology Institute,Chinan academy of Sciences.Mdck cell is purchased from
ATCC(CCL-34TM)。
It is commented using graded inhibition concentration index Fractional Inhibitory Concentration Index (FICI)
Valence nanometer selenium delivers Abiduoer delivery and inhibits H1N1 influenza virus inducing host cell apoptosis situations
Calculation formula is as follows：
Inhibition index=(SeNPs contents/SeNPs total amounts in Se@ARB nano particles)+(ARB in Se@ARB nano particles
Content/ARB total amounts)；
Carrying drug ratio (100%)=(drug carrier amount/drug total amount) × 100%；
FICI=(drug A combination/drug A alone)+(drug B combination/drug B
alone)
FICI ＜ 0.5 (collaboration)；0.5≤FICI≤2.0 (there was no significant difference)；FICI ＞ 2.0 (antagonism).
Elemental analysis (ICP-MS)：Measure SeNPs contents and SeNPs total amounts in Se@ARB nano particles；
Ultraviolet-visible absorption spectroscopy (UV-Vis)：Measure ARB contents and ARB total amounts in Se@ARB nano particles；
Inhibition index=(SeNPs contents/SeNPs total amounts in Se@ARB nano particles)+(ARB in Se@ARB nano particles
Content/ARB total amounts)
=(125 μM/1mM)+(5 μ g/mL/20 μ g/mL of content)
=0.125+0.25
=0.375
Embodiment 1
1) prepared by nanometer selenium delivery Abiduoer medicine-carried system
It is that 50mM vitamin c solutions are added dropwise to a concentration of 0.1M sodium selenites of 0.25mL to take 2mL stock solutions
(Na2SeO3) in solution, deionized water, which is added, makes final volume be 25mL.Magnetic agitation 2 hours, obtains after dialysis and receives after purification
Rice selenium solution, freeze-drying obtain nanometer selenium (SeNPs).It is 20mg/ to take above-mentioned nanometer selenium solution 10mL, 10 μ L storing solutions of addition
Abiduoer (ARB) solution of mL, magnetic agitation 1h, solution after reaction centrifuge 10 minutes by dialysis, 10000rpm, go from
Son washing three times, sample are freeze-dried to obtain nanometer selenium delivery Abiduoer nanometer medicine-carried system (Se@ARB).
Explore the optimisation substance ratio for preparing nanometer selenium delivery Abiduoer；Transmission electron microscope (TEM) observes nanometer selenium delivery
Abiduoer pattern changes；Elemental analysis (EDX) detects the element composition of nanometer selenium delivery Abiduoer.
Nanometer selenium delivers the preparation of Abiduoer and characterizes as shown in Figure 1.Figure A and B figures are the Tyndall effects of Se@ARB.
A and b is the transmission electron microscope picture of SeNPs and Se@ARB respectively in figure C, and as can be seen from the figure unmodified nanometer selenium is easy out
Now reunite, the nanometer selenium after functionalization is uniformly dispersed than independent nanometer selenium.Scheme D be elemental analysis, wherein Se elements account for 72%, C,
N and O is respectively 9%, 4% and 5% from ARB.Elemental analysis shows to obtain the nanometer selenium of functionalization.
2) nanometer selenium delivers Abiduoer to mdck cell toxicity test (MTT)
Steps are as follows：Single cell suspension is made with 0.25% trypsin digestion attached cell 3min, in 96 well culture plates
With 4 × 104A cells/well inoculation, be placed in incubator (37 DEG C, 5%CO2) preculture for 24 hours after, be added H1N1 virus (virus drop
Spend TCID50=103.5) it is incubated 2h, it is then added and contains various concentration medicine group (20 μ g/mL of ARB, 80 SeNPs μ g/mL, Se@
100 μ g/mL of ARB) each 100 μ L of culture medium, continue culture for 24 hours.Past culture plate addition 20 holes μ L/ MTT (5mg/mL), 37 DEG C
The liquid abandoned in cultivation plate hole is inhaled after being incubated 5h, 150 holes μ L/ DMSO are added, vibrates 10min, purple crystal thing fully dissolves
Afterwards, each hole OD is measured570Value.With control group OD570It is 100%, calculates drug-treated group cell survival rate.
Cell survival rate (%)=(OD570Experimental group/OD570Control group) × 100%.
Detect activity of the nanometer selenium delivery Abiduoer to cells following viral infection.It is (normal that mtt assay surveys different pharmaceutical component
Cell controls group, virus infected cell group, virus infected cell+nanometer selenium group (SeNPs80 μ g/mL), virus infected cell+Ah
It is more right than Duo Er group (20 μ g/mL of ARB), virus infected cell+nanometer selenium delivery Abiduoer group (100 μ g/mL of Se ARB)
Function and effect after mdck cell processing.After Fig. 2 shows virus infected cell, cell survival rate rate is 34%, relative to control group
It is decreased obviously.After virus infected cell, respectively plus after Abiduoer and nanometer selenium processing, cell survival rate is respectively 45% He
58%, risen relative to control group.After nanometer selenium delivers Abiduoer processing virus infected cell, survival rate rises to
86%, obviously rise relative to other components.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, it is other it is any without departing from the spirit and principles of the present invention made by changes, modifications, substitutions, combinations, simplifications,
Equivalent substitute mode is should be, is included within the scope of the present invention.
Claims (10)
1. a kind of preparation method of the Abiduoer functionalized nano selenium with anti-H1N1 influenza activities, it is characterised in that be
It is realized, is included the following steps using nanometer selenium load Abiduoer：
Abiduoer solution is added into nanometer selenium solution, dialyses, wash after being stirred to react, is dry, obtains nanometer selenium delivery
Abiduoer nanometer medicine-carried system, i.e. Abiduoer functionalized nano selenium.
2. the preparation side of the Abiduoer functionalized nano selenium according to claim 1 with anti-H1N1 influenza activities
Method, it is characterised in that：
In reaction system, the mass ratio of Abiduoer and nanometer selenium is 1: (2~8).
3. the system of the Abiduoer functionalized nano selenium according to claim 1 or 2 with anti-H1N1 influenza activities
Preparation Method, it is characterised in that：
In reaction system, the mass ratio of Abiduoer and nanometer selenium is 1: 4.
4. the preparation side of the Abiduoer functionalized nano selenium according to claim 1 with anti-H1N1 influenza activities
Method, it is characterised in that：
In reaction system, final concentration of 10~40 μ g/mL of Abiduoer.
5. the system of the Abiduoer functionalized nano selenium according to claim 1 or 4 with anti-H1N1 influenza activities
Preparation Method, it is characterised in that：
In reaction system, the final concentration of 20 μ g/mL of Abiduoer.
6. the preparation side of the Abiduoer functionalized nano selenium according to claim 1 with anti-H1N1 influenza activities
Method, it is characterised in that specifically comprise the following steps：
(1) 2mL 50mM vitamin c solutions is taken to be added dropwise in a concentration of 0.1M sodium selenite solutions of 0.25mL, addition go from
Sub- water makes final volume be 25mL, and magnetic agitation 2 hours obtains nanometer selenium solution after purification after dialysis；
(2) the nanometer selenium solution for taking step (1) to prepare, is added the Abiduoer solution of a concentration of 20mg/mL, makes in reaction system
The final concentration of Abiduoer reaches 10~40 μ g/mL；Magnetic agitation, the solution after reaction is by dialysis, centrifugation, deionization washing
Three times, sample is freeze-dried to obtain nanometer selenium delivery Abiduoer nanometer medicine-carried system, i.e. Abiduoer functionalized nano selenium.
7. the preparation side of the Abiduoer functionalized nano selenium according to claim 6 with anti-H1N1 influenza activities
Method, it is characterised in that：
The time of magnetic agitation described in step (2) is 1h.
8. a kind of Abiduoer functionalized nano selenium with anti-H1N1 influenza activities, it is characterised in that pass through claim
1~7 any one of them preparation method is prepared.
9. the Abiduoer functionalized nano selenium according to any one of claims 8 with anti-H1N1 influenza activities is preparing anti-H1N1
Application in influenza activity product.
10. application according to claim 9, it is characterised in that：
The effective concentration of the Abiduoer functionalized nano selenium is 80~100 μ g/mL.
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| CN201810382111.1A CN108514564A (en) | 2018-04-13 | 2018-04-13 | Abiduoer functionalized nano selenium with anti-H1N1 influenza activities and its preparation and application |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109091467A (en) * | 2018-09-12 | 2018-12-28 | 广州市妇女儿童医疗中心 | The nanometer selenium for loading Ribavirin inhibits the application in hand-foot-and-mouth disease viral product in preparation |
| CN110237093A (en) * | 2019-06-13 | 2019-09-17 | 广州市妇女儿童医疗中心 | The new opplication of sodium selenite |
| WO2023082213A1 (en) * | 2021-11-13 | 2023-05-19 | 暨南大学 | Novel coronavirus protein antigen nanovaccine, preparation method therefor and application thereof. |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102327620A (en) * | 2011-07-29 | 2012-01-25 | 暨南大学 | Application of nano-selenium in antineoplastic drug carrier |
-
2018
- 2018-04-13 CN CN201810382111.1A patent/CN108514564A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102327620A (en) * | 2011-07-29 | 2012-01-25 | 暨南大学 | Application of nano-selenium in antineoplastic drug carrier |
Non-Patent Citations (2)
| Title |
|---|
| LI ET AL.: ""Inhibition of H1N1 influenza virus-induced apoptosis by functionalized selenium nanoparticles with amantadine through ROSmediated AKT signaling pathways"", 《INTERNATIONAL JOURNAL OF NANOMEDICINE》 * |
| LI ET AL.: ""Inhibitory activity of selenium nanoparticles functionalized with oseltamivir on H1N1 influenza virus"", 《INTERNATIONAL JOURNAL OF NANOMEDICINE》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109091467A (en) * | 2018-09-12 | 2018-12-28 | 广州市妇女儿童医疗中心 | The nanometer selenium for loading Ribavirin inhibits the application in hand-foot-and-mouth disease viral product in preparation |
| CN110237093A (en) * | 2019-06-13 | 2019-09-17 | 广州市妇女儿童医疗中心 | The new opplication of sodium selenite |
| WO2023082213A1 (en) * | 2021-11-13 | 2023-05-19 | 暨南大学 | Novel coronavirus protein antigen nanovaccine, preparation method therefor and application thereof. |
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Application publication date: 20180911 |