CN109392900A - A kind of nano liposomes and its preparation method and application containing cell-penetrating peptides - Google Patents
A kind of nano liposomes and its preparation method and application containing cell-penetrating peptides Download PDFInfo
- Publication number
- CN109392900A CN109392900A CN201811249543.1A CN201811249543A CN109392900A CN 109392900 A CN109392900 A CN 109392900A CN 201811249543 A CN201811249543 A CN 201811249543A CN 109392900 A CN109392900 A CN 109392900A
- Authority
- CN
- China
- Prior art keywords
- cell
- penetrating peptides
- nano liposomes
- seq
- containing cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/34—Shaped forms, e.g. sheets, not provided for in any other sub-group of this main group
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
- A01N43/40—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom six-membered rings
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Pest Control & Pesticides (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Agronomy & Crop Science (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Organic Chemistry (AREA)
- Toxicology (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a kind of nano liposomes containing cell-penetrating peptides, including lecithin, surfactant, vitamin E and cell-penetrating peptides, and weight ratio is 10~60:10~30:0.1~10:0.2~20；Cell-penetrating peptides are selected from least one of biological source cell penetrating peptide, chimeric cell penetrating peptide and autosynthetic calls' penetrating peptide.In the preparation method of above-mentioned nano liposomes, using completely new high-pressure homogeneous dispersion method, nano liposomes stability is greatly improved, tank-mix adjuvants is agriculturally being used for or is flying anti-auxiliary agent.Nano liposomes stability provided by the invention containing cell-penetrating peptides is high, at normal temperature can long term storage, it is insensitive to pH value and ionic strength variation, it is good with pesticide and/or fertilizer compatibility；It is pollution-free without using organic solvent in preparation process；It can quickly be transmitted in vivo after use on crops, be rapidly reached Minimum effective concentration, extend efficiency time, improved control efficiency, drug resistance is delayed to generate.
Description
Technical field
The invention belongs to agriculture fields, contain the nano liposomes of cell-penetrating peptides and its preparation side more particularly to a kind of
Method, and it is somebody's turn to do the application of the nano liposomes containing cell-penetrating peptides.
Background technique
Nano liposomes are a kind of simulation membrane structure, made phospholipid bilayer structure.Nano liposomes
As a kind of pharmaceutical carrier that biocompatibility is splendid, it is widely used in the field of medicine.Nano liposomes can be taken
Band and embedding are hydrophilic, and hydrophobic and parents' class molecule is especially good to the molecule embedding effect that poorly water-soluble and grease are less soluble,
Its quick penetration organism surface can be assisted, and in its quickly distribution in vivo.Nano liposomes can be such that medicament is rapidly achieved
Minimum effective concentration, and effectively extend active drug concentration and hold time, enhance drug using effect, delays drug anti-
The generation of pharmacological property.But the main nano-lipid preparation such as film dispersion method reported at present, solvent injection method preparation
Nano liposomes require that pH stabilizer and cryo-conservation is added, and change sensitivity to ionic strength, show prepared by the above method
Nano liposomes be a dynamic equilibrium system, nano liposomes integrality height therein rely on solution buffer environment and
Temperature makes it impossible to agriculturally applying.
Cell-penetrating peptides are as a kind of polypeptide small molecule, and the amino acid number typically contained is less than 50.It can be helped
The vesica or sphere that nucleic acid, protein, virus, liposome and high molecular polymer are formed are contacted with cell outboard structure, and can
Autohemagglutination forms endocytosis and the pinocytosis for wearing mould complex or inducing cell, and allogenic material is assisted to enter cell.Into after cell,
It can induce the generation of escape intracellular, avoid allogenic material by lysosomal degradation.But cell-penetrating peptides cannot penetrate organism
The wax coat or cuticula on surface, therefore its independent and pesticide and/or effective fertilizer composition are with Covalent bonding together or mixing, Bu Nengbang
It helps it to enter in organism, plays its function of assisting allogene to enter cell.
In conclusion developing a kind of nano liposomes containing cell-penetrating peptides, stablize it under room temperature storage condition,
It is insensitive to pH value and ionic strength variation it is not necessary that pH stabilizer is added；Can wrap up and carry pesticide and/or fertilizer effectively at
Divide and enter in organism, and it is assisted to rapidly enter cell, enhance its using effect, the utilization of pesticide and fertilizer is greatly provided
Rate.
Summary of the invention
For the above the deficiencies in the prior art, the present invention provides a kind of nano liposomes containing cell-penetrating peptides, tools
Body is realized by the following technologies.
A kind of nano liposomes containing cell-penetrating peptides, including lecithin, surfactant, vitamin E and cell are worn
Saturating peptide, and weight ratio is 10~60:10~30:0.1~10:0.2~20；Wherein, it is thin to be selected from biological source for the cell-penetrating peptides
At least one of born of the same parents' penetrating peptide, chimeric cell penetrating peptide and autosynthetic calls' penetrating peptide.
Preferably, the weight ratio of the lecithin, surfactant, vitamin E and cell-penetrating peptides is 30:25:5:15.
Preferably, the cell-penetrating peptides be by HIV-1Tat, HIV-1Rev, Protamine 1, Human cJun,
At least one of Penetratin composition；
Wherein, the amino acid sequence of the HIV-1Tat is as shown in SEQ ID NO.1, the amino acid sequence of the HIV-1Rev
Column are as shown in SEQ ID NO.2, and the amino acid sequence of the Protamine 1 is as shown in SEQ ID NO.3, the Human
The amino acid sequence of cJun is as shown in SEQ ID NO.4, the amino acid sequence of the Penetratin such as SEQ ID NO.5 institute
Show.
Preferably, the cell-penetrating peptides are made of at least one of Transportan, MPG, Pep-1；
Wherein, the amino acid sequence of the Transportan is as shown in SEQ ID NO.6, the amino acid sequence of the MPG
As shown in SEQ ID NO.7, the amino acid sequence of the Pep-1 is as shown in SEQ ID NO.8.
Preferably, the cell-penetrating peptides are by least one of Polyarginines, Polylysines, MAP, R6W3
Composition；
Wherein, the amino acid sequence of the Polyarginines is as shown in SEQ ID NO.9, the Polylysines's
Amino acid sequence as shown in SEQ ID NO.10, the amino acid sequence of the MAP as shown in SEQ ID NO.11, the R6W3's
Amino acid sequence is as shown in SEQ ID NO.12.
Preferably, the lecithin is made of at least one of soybean lecithin, egg yolk lecithin, synthetic phospholipid.
It is highly preferred that the synthesis lecithin is by distearoylphosphatidylethanolamine-polyethylene glycol, two palmityl phosphatide
Acyl ethanol amine-polyethylene glycol, dioleoylphosphatidylethanolamine-polyethylene glycol, dilauroyl phosphatidyl-ethanolamine-poly- second two
At least one of alcohol composition, and the molecular weight of straight chain and/or branched chair polymacrogol contained in the synthesis lecithin be 200~
100000。
The present invention also provides the preparation methods of the above-mentioned nano liposomes containing cell-penetrating peptides, comprising the following steps:
S1, lecithin, surfactant, vitamin E, cell-penetrating peptides are weighed by weight, it is spare；
S2, surfactant and cell-penetrating peptides are added in reaction kettle, add surfactant and cell-penetrating peptides
The hot water that 50~200 times of total weight and temperature are 40~80 DEG C, stirring and dissolving obtain solution I；
S3, pure CO is passed through from reactor bottom2Air in replacement, removal reaction kettle；Then lecithin and vitamin E are taken
It adds in the solution I, while continuing to be passed through pure CO in reactor bottom2, make final pressure 0.3MPa in reaction kettle；Finally
It is rapidly heated to 125 DEG C of homogeneous 20min, until reaction solution is clarified；It stirs evenly and is cooled to room temperature normal pressure, obtain containing cell-penetrating peptides
The finished product of nano liposomes.
In above-mentioned preparation method, step S3 can be advanced optimized, specific prioritization scheme are as follows: firstly, to reactor bottom
It is passed through CO2Reacting kettle inner pressure is down to 0.05MPa using vacuum pump, then is passed through CO by 5min2It to normal pressure, recycles 5 times, removal
Air；Secondly, lecithin and vitamin E is taken to add in reaction kettle, stirred with the speed of 500r/min；Stirring is simultaneously in reaction kettle
High-purity CO is injected in bottom2, increase with 0.02MPa/min the pressure in reaction kettle, until stop when 0.3MPa, while boosting with
4 DEG C/min is rapidly heated to 125 DEG C；Then, homogenizer homogeneous 20min is opened with the speed of 30000r/min, until reaction solution
Clarification；Finally, temperature is down to 95 DEG C with 2.5 DEG C/min, and pressure is down to normal pressure with 0.02MPa/min in reaction kettle；Subsequent temperature
Room temperature is down to quickly to get the nano liposomes finished product containing cell-penetrating peptides with 2~4 DEG C/min again.
The above-mentioned nano liposomes containing cell-penetrating peptides provided by the present invention, can be applied in agriculture chemical field make
For a component in agricultural agent or agricultural agent, and application method are as follows: first by the nano-lipid containing cell-penetrating peptides
1:100~8000 is mixed by volume for body and water, and pesticide and/or fertilizer is added, application after being again stirring for uniformly.
Preferably, the above-mentioned nano liposomes containing cell-penetrating peptides can be used as tank-mix adjuvants, fly anti-auxiliary agent, or be bucket
One of mix auxiliary agent, fly anti-auxiliary agent.
Compared with prior art, the invention has the beneficial effects that:
1, the preparation method of the nano liposomes provided by the invention containing cell-penetrating peptides, i.e. high temperature, pressurization, homogeneous and
Inject CO2The method of common distribution makes lecithin and vitamin E be uniformly dispersed, and the liposome structure of formation is close, to extraneous item
Part variation is insensitive；
2, the nano liposomes stability as made from this method containing cell-penetrating peptides is high, can grow under normal temperature conditions
Time storage, it is insensitive to pH value and ionic strength variation, it is good with pesticide and/or fertilizer compatibility；
3, without using organic solvent, production technology safety and non-environmental-pollution problem in nano-lipid production procedure；
4, it in use, as tank-mix adjuvants, flying anti-auxiliary agent or tank-mix adjuvants, a kind of component for flying anti-auxiliary agent, can take
Band and hydrophilic, the hydrophobic and parents' class molecule of embedding, molecule especially less soluble to poorly water-soluble and grease；It can quick penetration
Organism surface and in vivo quickly distribution, are rapidly achieved Minimum effective concentration, and it is dense to greatly prolong active drug
Degree is held time, and is enhanced drug using effect, is delayed the generation of Drug Resistance；
5, the nano liposomes physicochemical property containing cell-penetrating peptides is excellent, dissipates what color method measured using dynamic Laser light
Average grain diameter is 129.5nm, and transmission electron microscope observing shows it with typical liposome；Its good biocompatibility, will not
Cause phytotoxicity or fertilizer damage, can be used in the entire breeding time of crop, will not dissolve and destroy plant surface wax coat structure.
Detailed description of the invention
Fig. 1 is transmission electricity of the nano liposomes containing cell-penetrating peptides of the preparation of embodiment 1 after phosphotungstic acid negative staining
Mirror picture；
Fig. 2 is that the nano liposomes containing cell-penetrating peptides prepared by embodiment 1 dissipate color method survey by dynamic Laser light
Obtain particle diameter distribution situation；
Specific embodiment
Below in conjunction with the attached drawing in the present invention, technical solution of the present invention is clearly and completely described, it is clear that
Described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on the implementation in the present invention
Example, those of ordinary skill in the art's all other embodiment obtained under the conditions of not making creative work belong to
The scope of protection of the invention.
In following embodiment, the nano liposomes containing cell-penetrating peptides the preparation method is as follows:
P1, by the weight demands of each embodiment, weigh lecithin, surfactant, vitamin E, cell-penetrating peptides, it is standby
With；
P2, surfactant and cell-penetrating peptides are added in reaction kettle, add 55 DEG C of hot water；Add in each embodiment
The weight for entering hot water is M1, stirring and dissolving obtains solution I；
P3, CO is passed through to reactor bottom2Reacting kettle inner pressure is down to 0.05MPa using vacuum pump, then led to by 5min
Enter CO2It to normal pressure, recycles 5 times, removes air；
P4, it takes lecithin and vitamin E to add in reaction kettle, is stirred with the speed of 500r/min；Simultaneously in reaction kettle bottom
High-purity CO is injected in portion2, increase with 0.02MPa/min the pressure in reaction kettle, until stop when 0.3MPa, with 4 while boosting
DEG C/min is rapidly heated to 125 DEG C；
P5, homogenizer homogeneous 20min is opened with the speed of 30000r/min, until reaction solution is clarified；
In P6, reaction kettle, temperature is down to 95 DEG C with 2.5 DEG C/min, and pressure is down to normal pressure with 0.02MPa/min；Then temperature
Degree is down to room temperature quickly with 2~4 DEG C/min again to get the nano liposomes finished product containing cell-penetrating peptides.
Examples 1 to 3
Contain the nano liposomes of cell-penetrating peptides, the weight and preparation step of raw material described in Examples 1 to 3
The weight of hot water is added in P2, as shown in table 1 below.
The weight of hot water, unit g in the raw material and step P2 of 1 Examples 1 to 3 of table
Application examples 1: Electronic Speculum detection is carried out to the nano liposomes containing cell-penetrating peptides prepared by embodiment 1
Nano liposomes containing cell-penetrating peptides are added drop-wise on the copper mesh that transmission electron microscope uses, the phosphorus of 1 drop 1% is added dropwise
Wolframic acid.It is detected after drying at room temperature.
Testing result as shown in Figure 1, obtain the nano liposomes partial size containing cell-penetrating peptides 100~200nm it
Between, surface has irregular protrusion, matches with the structure feature of liposome.
Application examples 2: to the droplet measurement of the nano liposomes containing cell-penetrating peptides prepared by embodiment 1
The nano liposomes for taking 1ml to contain cell-penetrating peptides use Nano-ZS nano particle size and the Zeta electricity of Malvern
Its average grain diameter of position analysis-e/or determining.
Testing result is as shown in Fig. 2, the average grain diameter of the nano liposomes containing cell-penetrating peptides obtained is
129.5nm, PdI value are 0.128, show that the nano liposomes uniformity containing cell-penetrating peptides obtained is preferable, quality is higher.
Application examples 3: the Stability Determination of the nano liposomes containing cell-penetrating peptides
By the nano liposomes containing cell-penetrating peptides of Examples 1 to 3 preparation and using conventional ethanol injection legal system
The standby nano liposomes containing cell-penetrating peptides, respectively test ordinary temperature stability (30 DEG C, 90d), high-temperature stability (54 DEG C,
3d), low-temperature stability (4 DEG C, 3d), pH stability, ion stability.As a result as shown in table 2 below.
Nano liposomes stability of the table 2 containing cell-penetrating peptides
*: the nano liposomes containing cell-penetrating peptides being mixed with the HCl solution of pH=3 by 1:100, are seen after standing 4h
It examines；
*: the nano liposomes containing cell-penetrating peptides are mixed with the NaOH solution of pH=10 by 1:100, after standing 4h
Observation；
* * is by the nano liposomes containing cell-penetrating peptides and the hard water (MgCl2 of the CaCl2 of 0.3g/L, 0.15g/L
It 6H2O) mixes by 1:100, is observed after standing 4h；
As shown in Table 2, under various conditions using the nano liposomes made from Examples 1 to 3 containing cell-penetrating peptides
Clarification is kept, and the nano liposomes as made from conventional ethanol injection method containing cell-penetrating peptides are only in 4 DEG C of low temperature items
It keeps stablizing under part, occurs precipitating under the conditions of other or muddy situation.Show high-pressure homogeneous dispersion method in preparation containing thin
There is unique advantage in the nano liposomes of born of the same parents' penetrating peptide.
Using 4: the nano liposomes for containing cell-penetrating peptides of Examples 1 to 3 preparation improve fluorine pyridine as tank-mix adjuvants
Control efficiency of the insect amide to planthopper
Using 5 methods, the insect population of each experimental plot is investigated.Examples 1 to 3 preparation is contained into cell-penetrating peptides
Nano liposomes and clear water press 1:1,500 stir evenly, and configuration flonicamid effective ingredient consumption is 5g/ mus of medical fluid,
In addition the nano liposomes containing cell-penetrating peptides and clear water that prepared by Example 1 press 1:1, and 500 stir evenly, and configure fluorine pyridine
The medical fluid that insect amide effective ingredient consumption is 4g, 3g/ mu.As control, using clear water be respectively configured flonicamid effectively at
Dividing dosage is 5g, 4g, 3g/ mu of medical fluid.Clear water processing control is set simultaneously, and 4 repetitions are arranged in each processing.Respectively in medication
Each hillslope processes population number is investigated after 3,7,14 and 21 days afterwards, calculates control efficiency.As a result as shown in table 3 below
Control efficiency of 3 flonicamid of table to planthopper
As shown in table 3, by comparative example 1 (1), 2,3 and 1 group of control, be added to Examples 1 to 3 contains cell
The flonicamid 5g a.i. of the nano liposomes of penetrating peptide/mu processing preventive effect is apparently higher than and is not added with containing cell-penetrating peptides
Nano liposomes flonicamid 5g a.i./mu processing 10%.Pass through comparative example 1 (2), 1 (3) and control 2,3
Group, each processing preventive effect of flonicamid for being added to the nano liposomes containing cell-penetrating peptides of embodiment 1 are above not
The flonicamid processing 10~20% for adding the nano liposomes containing cell-penetrating peptides shows that it has quickly conduction and protects
Protect the effect of effective component.Meanwhile wherein 14 days and 21 days synergy amplitudes are maximum, show that it has sustained release to effective component
Effect.
It can be seen that the control efficiency of flonicamid can be improved in the nano liposomes containing cell-penetrating peptides, extend
Its lasting period.
Sequence table
<110>Wuhan Kang Ke plant protection technology Co., Ltd
<120>a kind of nano liposomes and its preparation method and application containing cell-penetrating peptides
<141> 2018-10-25
<160> 12
<170> SIPOSequenceListing 1.0
<210> 1
<211> 9
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 1
Arg Lys Lys Arg Arg Gln Arg Arg Arg
1 5
<210> 2
<211> 17
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 2
Thr Arg Gln Ala Arg Arg Asn Arg Arg Arg Arg Trp Arg Glu Arg Gln
1 5 10 15
Arg
<210> 3
<211> 32
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 3
Pro Arg Arg Arg Arg Ser Ser Ser Arg Pro Val Arg Arg Arg Arg Arg
1 5 10 15
Pro Arg Val Ser Arg Arg Arg Arg Arg Arg Gly Gly Arg Arg Arg Arg
20 25 30
<210> 4
<211> 28
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 4
Arg Ile Lys Ala Glu Arg Lys Arg Met Arg Asn Arg Ile Ala Ala Ser
1 5 10 15
Lys Ser Arg Lys Arg Lys Leu Glu Arg Ile Ala Arg
20 25
<210> 5
<211> 16
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 5
Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys
1 5 10 15
<210> 6
<211> 27
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 6
Gly Trp Thr Leu Asn Ser Ala Gly Tyr Leu Leu Gly Lys Ile Asn Leu
1 5 10 15
Lys Ala Leu Ala Ala Leu Ala Lys Lys Ile Leu
20 25
<210> 7
<211> 27
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 7
Gly Ala Leu Phe Leu Gly Phe Leu Gly Ala Ala Gly Ser Thr Met Gly
1 5 10 15
Ala Trp Ser Gln Pro Lys Lys Lys Arg Lys Val
20 25
<210> 8
<211> 21
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 8
Lys Glu Thr Trp Trp Glu Thr Trp Trp Thr Glu Trp Ser Gln Pro Lys
1 5 10 15
Lys Lys Arg Lys Val
20
<210> 9
<211> 35
<212> PRT
<213>artificial sequence (Artificial Sequence)
<220>
<221> MOD_RES
<222> (1)..(35)
<223> This region may encompass 6 to 20 Arg residues
<400> 9
Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg
1 5 10 15
Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg
20 25 30
Arg Arg Arg
35
<210> 10
<211> 35
<212> PRT
<213>artificial sequence (Artificial Sequence)
<220>
<221> MOD_RES
<222> (1)..(35)
<223> This region may encompass 6 to 20 Lys residues
<400> 10
Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys
1 5 10 15
Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys
20 25 30
Lys Lys Lys
35
<210> 11
<211> 18
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 11
Lys Leu Ala Leu Lys Leu Ala Leu Lys Ala Leu Lys Ala Ala Leu Lys
1 5 10 15
Leu Ala
<210> 12
<211> 9
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 12
Arg Arg Trp Trp Arg Arg Trp Arg Arg
1 5
Claims (10)
1. a kind of nano liposomes containing cell-penetrating peptides, which is characterized in that including lecithin, surfactant, vitamin E
And cell-penetrating peptides, and weight ratio is 10~60:10~30:0.1~10:0.2~20；Wherein, the cell-penetrating peptides are selected from
At least one of biological source cell penetrating peptide, chimeric cell penetrating peptide and autosynthetic calls' penetrating peptide.
2. the nano liposomes according to claim 1 containing cell-penetrating peptides, which is characterized in that the lecithin, table
The weight ratio of face activating agent, vitamin E and cell-penetrating peptides is 30:25:5:15.
3. the nano liposomes according to claim 1 or 2 containing cell-penetrating peptides, which is characterized in that the cell is worn
Saturating peptide is made of at least one of HIV-1Tat, HIV-1Rev, Protamine 1, Human cJun, Penetratin；
Wherein, the amino acid sequence of the HIV-1Tat is as shown in SEQ ID NO.1, and the amino acid sequence of the HIV-1Rev is such as
Shown in SEQ ID NO.2, the amino acid sequence of the Protamine 1 as shown in SEQ ID NO.3, the Human cJun's
Amino acid sequence is as shown in SEQ ID NO.4, and the amino acid sequence of the Penetratin is as shown in SEQ ID NO.5.
4. the nano liposomes according to claim 1 or 2 containing cell-penetrating peptides, which is characterized in that the cell is worn
Saturating peptide is made of at least one of Transportan, MPG, Pep-1；
Wherein, the amino acid sequence of the Transportan is as shown in SEQ ID NO.6, and the amino acid sequence of the MPG is such as
Shown in SEQ ID NO.7, the amino acid sequence of the Pep-1 is as shown in SEQ ID NO.8.
5. the nano liposomes according to claim 1 or 2 containing cell-penetrating peptides, which is characterized in that the cell is worn
Saturating peptide is made of at least one of Polyarginines, Polylysines, MAP, R6W3；
Wherein, the amino acid sequence of the Polyarginines is as shown in SEQ ID NO.9, the amino of the Polylysines
Acid sequence is as shown in SEQ ID NO.10, and the amino acid sequence of the MAP is as shown in SEQ ID NO.11, the amino of the R6W3
Acid sequence is as shown in SEQ ID NO.12.
6. the nano liposomes according to claim 1 or 2 containing cell-penetrating peptides, which is characterized in that the lecithin
It is made of at least one of soybean lecithin, egg yolk lecithin, synthetic phospholipid.
7. the nano liposomes according to claim 6 containing cell-penetrating peptides, which is characterized in that the synthesis lecithin
By distearoylphosphatidylethanolamine-polyethylene glycol, dipalmitoylphosphatidylethanolamine-polyethylene glycol, dioleoyl phosphatide
At least one of acyl ethanol amine-polyethylene glycol, dilauroyl phosphatidyl-ethanolamine-polyethylene glycol composition, and the synthesis ovum
The molecular weight of straight chain and/or branched chair polymacrogol contained in phosphatide is 200~100000.
8. the preparation method of the nano liposomes containing cell-penetrating peptides described in claim 1,2 or 7, which is characterized in that packet
Include following steps:
S1, lecithin, surfactant, vitamin E, cell-penetrating peptides are weighed by weight, it is spare；
S2, surfactant and cell-penetrating peptides are added in reaction kettle, add surfactant and cell-penetrating peptides gross weight
The hot water that 50~200 times of amount and temperature are 40~80 DEG C, stirring and dissolving obtain solution I；
S3, pure CO is passed through from reactor bottom2Air in replacement, removal reaction kettle；Then lecithin and vitamin E is taken to add to
In the solution I, while continuing to be passed through pure CO in reactor bottom2, make final pressure 0.3MPa in reaction kettle；It is last quick
125 DEG C of homogeneous 20min are warming up to, until reaction solution is clarified；It stirs evenly and is cooled to room temperature normal pressure, obtain the nanometer containing cell-penetrating peptides
The finished product of liposome.
9. the application of the nano liposomes containing cell-penetrating peptides described in claim 1,2 or 7, which is characterized in that be applied to
A component being used as in agriculture chemical field in agricultural agent or agricultural agent, and application method are as follows: will first contain cell
1:100~8000 is mixed the nano liposomes and water of penetrating peptide by volume, and pesticide and/or fertilizer is added, is again stirring for uniformly
After apply.
10. the application of the nano liposomes pair according to claim 9 containing cell-penetrating peptides, which is characterized in that described
Agricultural agent is tank-mix adjuvants or flies anti-auxiliary agent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811249543.1A CN109392900B (en) | 2018-10-25 | 2018-10-25 | Nano liposome containing cell penetrating peptide and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811249543.1A CN109392900B (en) | 2018-10-25 | 2018-10-25 | Nano liposome containing cell penetrating peptide and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109392900A true CN109392900A (en) | 2019-03-01 |
| CN109392900B CN109392900B (en) | 2020-05-19 |
Family
ID=65469209
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811249543.1A Active CN109392900B (en) | 2018-10-25 | 2018-10-25 | Nano liposome containing cell penetrating peptide and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109392900B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109874788A (en) * | 2019-03-05 | 2019-06-14 | 烟台大学 | A kind of protein nano Liposomal formulation and its preparing the application in biological nano plant growth regulator |
| CN110037025A (en) * | 2019-05-10 | 2019-07-23 | 临沂伯特利生物科技有限公司 | A kind of environment-friendly type fruit tree antifreezing agent and preparation method thereof |
| CN110201147A (en) * | 2019-05-23 | 2019-09-06 | 中山市粤美医疗生物科技有限公司 | A kind of composition and preparation method thereof comprising peptide |
| CN110742064A (en) * | 2019-11-05 | 2020-02-04 | 江苏擎宇化工科技有限公司 | Modified liposome and composition, preparation method and application thereof |
| CN112450211A (en) * | 2019-09-06 | 2021-03-09 | 南京市高淳区红太阳农资配送有限公司 | Agricultural tank-mixed spray synergist and preparation method thereof |
| CN113693086A (en) * | 2021-09-08 | 2021-11-26 | 山西省农业科学院园艺研究所 | Drought-resistant agent special for iris plants |
| CN115868494A (en) * | 2022-08-08 | 2023-03-31 | 武汉康科植保技术有限公司 | Fragrant rice flavor enhancer containing cell penetrating peptide and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103876982A (en) * | 2014-03-24 | 2014-06-25 | 烟台新时代健康产业日化有限公司 | Nano-liposome emulsion and preparation method thereof |
| CN107446016A (en) * | 2016-05-30 | 2017-12-08 | 上海交通大学 | A kind of cell-penetrating peptide of Stearate Modified and its preparation and application |
| CN108135846A (en) * | 2015-10-21 | 2018-06-08 | 海德堡大学 | For the liposome containing cell-penetrating peptides and tetraether lipid of oral delivery macromolecular |
-
2018
- 2018-10-25 CN CN201811249543.1A patent/CN109392900B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103876982A (en) * | 2014-03-24 | 2014-06-25 | 烟台新时代健康产业日化有限公司 | Nano-liposome emulsion and preparation method thereof |
| CN108135846A (en) * | 2015-10-21 | 2018-06-08 | 海德堡大学 | For the liposome containing cell-penetrating peptides and tetraether lipid of oral delivery macromolecular |
| CN107446016A (en) * | 2016-05-30 | 2017-12-08 | 上海交通大学 | A kind of cell-penetrating peptide of Stearate Modified and its preparation and application |
Non-Patent Citations (2)
| Title |
|---|
| RAJIV KUMAR等: "Lecithin Organogels as a Potential Phospholipid-Structured System for Topical Drug Delivery: A Review", 《AAPS PHARMSCITECH》 * |
| 张强、武凤兰主编: "《药剂学》", 31 January 2005, 北京大学医学出版社 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109874788A (en) * | 2019-03-05 | 2019-06-14 | 烟台大学 | A kind of protein nano Liposomal formulation and its preparing the application in biological nano plant growth regulator |
| CN110037025A (en) * | 2019-05-10 | 2019-07-23 | 临沂伯特利生物科技有限公司 | A kind of environment-friendly type fruit tree antifreezing agent and preparation method thereof |
| CN110037025B (en) * | 2019-05-10 | 2020-09-11 | 临沂伯特利生物科技有限公司 | Environment-friendly fruit tree antifreezing agent and preparation method thereof |
| CN110201147A (en) * | 2019-05-23 | 2019-09-06 | 中山市粤美医疗生物科技有限公司 | A kind of composition and preparation method thereof comprising peptide |
| CN112450211A (en) * | 2019-09-06 | 2021-03-09 | 南京市高淳区红太阳农资配送有限公司 | Agricultural tank-mixed spray synergist and preparation method thereof |
| CN110742064A (en) * | 2019-11-05 | 2020-02-04 | 江苏擎宇化工科技有限公司 | Modified liposome and composition, preparation method and application thereof |
| CN110742064B (en) * | 2019-11-05 | 2021-07-20 | 江苏擎宇化工科技有限公司 | Modified liposome and composition, preparation method and application thereof |
| CN113693086A (en) * | 2021-09-08 | 2021-11-26 | 山西省农业科学院园艺研究所 | Drought-resistant agent special for iris plants |
| CN115868494A (en) * | 2022-08-08 | 2023-03-31 | 武汉康科植保技术有限公司 | Fragrant rice flavor enhancer containing cell penetrating peptide and preparation method and application thereof |
| CN115868494B (en) * | 2022-08-08 | 2023-12-26 | 武汉康科植保技术有限公司 | Fragrant rice flavoring agent containing cell penetrating peptide, and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109392900B (en) | 2020-05-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109392900A (en) | A kind of nano liposomes and its preparation method and application containing cell-penetrating peptides | |
| US20220304903A1 (en) | Method of preparing bioactive substance-encapsulated ethosome, ethosome composition, and cosmetic composition including ethosome composition | |
| EP3315138B1 (en) | Stick-shaped substrate containing lipid-peptide-type compound | |
| CN107920964A (en) | Merge the coated porous silicon nanoparticles of liposome | |
| CN110237035B (en) | Active targeting amphiphilic polypeptide nano-drug carrier and preparation and application thereof | |
| CN101524331B (en) | Polysaccharide-liposome and preparation method and purpose thereof | |
| CN105148265B (en) | The peptide formulations of carbon fluorine connection | |
| CN107096012A (en) | Composition of peptide and preparation method thereof | |
| CN107595809A (en) | A kind of zeins nanometer embedding sustained release filler and preparation method thereof | |
| KR20170117438A (en) | A transdermal absorption device containing a lipid peptide-type compound | |
| CN105521494A (en) | Cell penetrating peptide introduced drug-delivery carrier comprising macromolecule | |
| CA2425271C (en) | Liposomes encapsulating anticancerous drugs and use thereof in the treatment of malignant tumours | |
| US20110020428A1 (en) | Gel-stabilized liposome compositions, methods for their preparation and uses thereof | |
| KR20170002431A (en) | Thickening composition containing lipid peptide-type compound | |
| KR102190935B1 (en) | Nano-lipid carrier for encapsulation of physiologically active substance and preparation method thereof | |
| CN111607093A (en) | pH sensitive nano-carrier and application thereof in gene drug delivery | |
| CN107708660A (en) | Include the bar-shaped base material of lipid peptide type compound | |
| CN105832702B (en) | For the protein nano magnetic crust core capsule of drug delivery and its application | |
| CN108367094A (en) | Peptide combinations | |
| EP3560480A1 (en) | Stick-type solid preparation base for external application to skin | |
| JP2022527568A (en) | Lipid-encapsulated amphipathic peptide | |
| Lemma et al. | Disruption of giant unilamellar vesicles mimicking cell membranes induced by the pesticides glyphosate and picloram | |
| TW202042842A (en) | Topical preparation for the skin or mucosae, method for manufacturing same, and base for topical preparation for the skin or mucosae | |
| DE60025332T2 (en) | SYNTHETIC PEPTIDE FOR TROUBLESHOOTING INTRA-CELLULAR SIGNALS | |
| KR101837023B1 (en) | Cosmetic nanoliposome composition containg peptides for inhibiting degradation of collagen and manufacturing method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |