CN105801668A - Oligoarginine modified phospholipid, nanoparticles assembled by oligoarginine modified phospholipid, preparation method of oligoarginine modified phospholipid and application of nanoparticles - Google Patents
Oligoarginine modified phospholipid, nanoparticles assembled by oligoarginine modified phospholipid, preparation method of oligoarginine modified phospholipid and application of nanoparticles Download PDFInfo
- Publication number
- CN105801668A CN105801668A CN201610207923.3A CN201610207923A CN105801668A CN 105801668 A CN105801668 A CN 105801668A CN 201610207923 A CN201610207923 A CN 201610207923A CN 105801668 A CN105801668 A CN 105801668A
- Authority
- CN
- China
- Prior art keywords
- modified
- phospholipid
- arginine
- oligomerization
- oligoarginine
- 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
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- 150000003904 phospholipids Chemical class 0.000 title claims abstract description 55
- 239000002105 nanoparticle Chemical class 0.000 title claims abstract description 54
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- -1 NHS activated phosphatidic acid Chemical class 0.000 claims abstract description 20
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- 229940079593 drug Drugs 0.000 claims abstract description 16
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 claims abstract description 14
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 13
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 claims abstract description 12
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- 239000004475 Arginine Substances 0.000 claims description 72
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 72
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- PORPENFLTBBHSG-MGBGTMOVSA-N 1,2-dihexadecanoyl-sn-glycerol-3-phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(O)=O)OC(=O)CCCCCCCCCCCCCCC PORPENFLTBBHSG-MGBGTMOVSA-N 0.000 claims description 10
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- NXLNNXIXOYSCMB-UHFFFAOYSA-N (4-nitrophenyl) carbonochloridate Chemical compound [O-][N+](=O)C1=CC=C(OC(Cl)=O)C=C1 NXLNNXIXOYSCMB-UHFFFAOYSA-N 0.000 claims description 3
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/10—Tetrapeptides
- C07K5/1019—Tetrapeptides with the first amino acid being basic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
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Abstract
The invention relates to oligoarginine modified phospholipid, nanoparticles assembled by the oligoarginine modified phospholipid, a preparation method of the oligoarginine modified phospholipid and an application of the nanoparticles. After oligoarginine peptide chains are activated by DCC (dicyclohexylcarbodiimide)/NHS (N-hydroxysuccinimide), the oligoarginine peptide chains react with phosphatidyl ethanolamine, purification is performed, and oligoarginine modified phosphatidyl ethanolamine is obtained; or the oligoarginine peptide chains react with DCC/NHS activated phosphatidic acid, and oligoarginine modified phosphatidic acid is obtained through purification; or the oligoarginine peptide chains react with nitrophenyl chloroformate activated phosphatidylcholine, and oligoarginine modified phosphatidylcholine is obtained through purification. The nanoparticles assembled by the oligoarginine modified phospholipid is used for supporting genes, small-interfering RNA, polypeptides or proteinic drugs, antibodies and chemical drugs, and the formed aqueous dispersion of the drug-carrying nanoparticles is used for delivering drugs for in-vitro cells or in-vivo local intravenous injection. The nanoparticles are effectively promoted to enter the cells, and the intracellular drug delivery efficiency is improved.
Description
Technical field
The present invention relates to the nanoparticle that assembles with it of phospholipid and preparation method and application that a kind of oligomerization arginine is modified, feature is this
The phospholipid molecule that oligomerization arginine is modified has the nanoparticle entrance cell that mediation is assembled, and promotes medicine intracellular delivery efficiency
Function, belongs to nano-carrier technical field.
Background technology
The medicines such as chemicals, gene, albumen, polypeptide and antibody, it usually needs by vehicle delivery to intracellular
Wave effect.Although viral vector has a higher transfection efficiency, but owing to self immunogenicity and biological safety are asked
Topic, limits their application.Nano-carrier has vast potential for future development at drug delivery field, is targeted drug
The hope delivered.But the obstacles such as cell membrane, and the problem such as drug resistance, make the high potency drugs in target cell deliver, especially
It is that the medicine deliverys such as gene, nucleic acid, polypeptide, antibody there is also huge challenge.Non-virus carrier delivery of gene,
The medicines such as polypeptide, it usually needs enter intracellular endosome and lysosome through cell endocytic approach, need through interior
Contain body escape and discharge loaded medicine.Due to endosome and lysosomal sour environment and the effect of various enzyme,
Often make the biological species drug degradation such as gene, nucleic acid, be substantially reduced delivery efficiency.Research shows, nano-carrier warp
Endocytosis enters after endosome, the endosome slip time window of only 5～15 minutes, otherwise enter endosome in late period or with
After lysosome fusion, then it is difficult to escape out again and discharge entrained small nucleic acids molecule (siRNA).Even at present
The preferable lipid nanoparticle of effect, is only capable of escaping from lysosome and discharging 3.5% after entering cellular inclusion
siRNA.To this end, we it may be necessary design novel nano carrier to solve the thin of the biological species medicine such as gene, polypeptide
Intracellular delivery problems.
Summary of the invention
It is an object of the invention to provide a kind of efficiently mediation carrier and enter cell, in promoting the drug cells such as gene, deliver merit
The technological approaches of effect, is i.e. divided by the carbochain phospholipid of a kind of oligomerization arginine modification with cell membrane with the strongest fusion faculty
Son, mediates the lipid nanoparticle assembled altogether by it through medicines such as cell membrane delivery of gene, siRNA；Including oligomerization essence ammonia
Phospholipid (hereinafter referred to as CP-A) that acid is modified and preparation method thereof, CP-A assembles jointly with carbochain-Polyethylene Glycol and cholesterol
Lipid nanoparticle and assemble method and application.
The present invention is realized by the following technical programs:
The phospholipid that the oligomerization arginine that the present invention relates to is modified, wherein, oligomerization arginine is containing 4-8 arginine unit
Oligomerization arginine, modifies on the phosphoric acid head base of phospholipid.
Described phospholipid selects PHOSPHATIDYL ETHANOLAMINE, phosphatidylcholine, phosphatidyl glycerol, phosphatidic acid, Phosphatidylserine
Or phosphatidylinositols.
Described phospholipid, is optimized for the phospholipid that hydrocarbon chain contains 12-20 carbon atom further.
The phospholipid that described oligomerization arginine is modified, wherein oligomerization arginine is optimized for containing 4 or 5 arginine unit,
Phospholipid at least contains 2 hydrocarbon chains.
Phospholipid of the present invention preferably is selected from dilauroyl PHOSPHATIDYL ETHANOLAMINE (DLPE), two myristoyl phosphatidyls
Ethanolamine (DMPE), DPPE (DPPE), DSPE (DSPE),
Two Semen arachidis hypogaeae acylphosphatidyl ethanolamines (DAPE), DOPE (DOPE), two myristoyl phospholipid
Acyl glycerol (DMPG), DSPG (DSPG), DLPG (DLPG), two
Palmityl phosphatidyl glycerol (DPPG), DOPG (DOPG), EPG (EPG), two oil
Acyl phosphatidic acid (DOPA), two myristoyl phosphatidic acid (DMPA), DPPA (DPPA), distearyl acyl group phosphorus
Fat acid (DSPA), dilauroyl phosphatidic acid (DLPA), dilauroyl phosphatidyl acid (DLPA), two Semen Myristicaes
Phosphatidyl choline (DMPC), DPPE (DPPA), DSPC
(DSPC), two Semen arachidis hypogaeae phosphatidyl choline (DAPC) or dioleyl phosphatidyl choline (DOPC).
The phospholipid that oligomerization arginine of the present invention is modified, is prepared via a method which: oligomerization arginine peptide chain is used
After dicyclohexylcarbodiimide (DCC)/N-hydroxy-succinamide (NHS) activation, react with PHOSPHATIDYL ETHANOLAMINE, pass through
Preparation liquid phase is purified and obtains the PHOSPHATIDYL ETHANOLAMINE that oligomerization arginine is modified；Or oligomerization arginine peptide chain bicyclo-
After hexyl carbodiimide/DMAP activation, react with phosphatidyl glycerol, obtain oligomerization arginine through purification
The phosphatidyl glycerol modified；Or the phosphatidic acid that oligomerization arginine peptide chain and DCC/NHS activate reacts, through preparing liquid
It is purified mutually and obtains the phosphatidic acid that oligomerization arginine is modified；Or oligomerization arginine peptide chain and p-nitrophenyl chloro-carbonic acid
The phosphatidylcholine reaction of ester activation, is purified through preparation liquid phase and obtains the phosphatidylcholine that oligomerization arginine is modified.
The phospholipid modified with oligomerization arginine of the present invention, assembles jointly with other lipid, forms 30～200nm
Lipid nanoparticle or liposome, wherein oligomerization arginine modify phospholipid in nanoparticle weight/mass percentage composition be
5%～80%.
The nanoparticle that the phospholipid that described oligomerization arginine is modified is assembled, the phosphorus that further preferred oligomerization arginine is modified
Fat assembles the nanoparticle of formation altogether with phosphatidylcholine, cholesterol and polyethyleneglycol modified lipid.Wherein oligomerization essence ammonia
Acid mass content is 10%～80%, and phosphatidylcholine mass content is 1%～50%, cholesterol mass content 5%～50%,
DSPE-PEG mass content is 0.5%～10%.
The nanoparticle that the phospholipid that described oligomerization arginine is modified is assembled, is the widow containing 4 or 5 arginine unit
The phospholipid that poly arginine is modified and cholesterol, DOPC, polyethyleneglycol modified DSPE
(DSPE-PEG) nanoparticle formed, the wherein molecular weight of PEG preferably 1000～5000 are assembled.
The nanoparticle that the phospholipid that the oligomerization arginine of the present invention is modified is assembled, can be prepared via a method which load water soluble
Or fat-soluble medicine: utilize ultrasonic dispersion, first water soluble drug is dissolved in phosphate buffer, selected various phospholipid,
Cholesterol and fat-soluble medicine are codissolved in organic solvent, stir evaporating organic solvent, and residual liquid, through ultrasonic Treatment, is then peeled off
Go out liposome, then be suspended in phosphate buffer, make liposome suspension type injection.
The phospholipid that oligomerization arginine in the present invention is modified, it is also possible to receive with assembling formation altogether such as other lipid, polymer etc.
The grain of rice.
The nanoparticle that the phospholipid that described oligomerization arginine is modified is assembled, can be used for loading gene, siRNA, many
Peptide or protein medicaments, antibody and chemicals, the aqueous dispersions of the drug-carrying nanometer particle of formation, for cell in vitro
Or internal local, intravenous injection deliver medicine.Feature is can to effectively facilitate nanoparticle to enter cell, improves intracellular medicine and passs
Send efficiency.
Accompanying drawing explanation
The structure of the DSPE (DSPE-A4) that the oligomerization arginine (A4) of preparation is modified in Fig. 1 embodiment 1
Schematic diagram and mass spectra peak.1630 occur unimodal, it was demonstrated that 4 poly arginines are bonded on DSPE.
In Fig. 2 embodiment 28, DSPE-A4 and cholesterol, DOPC, DSPE-PEG assemble the nanoparticle of formation
Grain size distribution.
In Fig. 3 embodiment 28, DSPE-A4 and cholesterol, DOPC, DSPE-PEG assemble the saturating of the nanoparticle of formation
Radio mirror (TEM) photo.
The gel electrophoresis experiment knot of load DNA nanoparticle (NP-29, NP-30 and NP-31) prepared by Fig. 4 embodiment 29-31
Really, blank NP-28 nanoparticle mixes (NP-28+DNA) group with DNA can not load upper DNA, and NP-29, NP-30
All can DNA in payload with NP-31.
Fig. 5 loads DNA nanoparticle NP-29 in-vitro transfection experimental result, and the transfection of NP-29 gathers apparently higher than conventional
Aziridine (PEI) carrier.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described in detail.Following example will assist in those skilled in the art and enter one
Step understands the present invention, but limits the present invention the most in any form.It is pointed out that to those skilled in the art,
Without departing from the inventive concept of the premise, it is also possible to make some deformation and improvement.These broadly fall into protection scope of the present invention.
Oligomerization arginine used is bought from Shanghai Tao Pu polypeptide, arginic 4-8 aggressiveness be expressed as A4, A5, A6, A7,
A8；The phospholipid etc. that used various phospholipid, Polyethylene Glycol (PEG) are modified the most directly buys commercially available prod.
Embodiment 1:
The preparation of the DSPE (DSPE-A4) that the arginine tetramer (A4) is modified: 1mol Amino End Group second
The A4 of acid blocked, is dissolved in DMSO solution 1ml, adds 0.4g dicyclohexylcarbodiimide (DCC) DCC and 0.4g/N-
N-Hydroxysuccinimide (NHS), activates 1 hour by 25 DEG C, is subsequently adding DSPE 1mol (purchased from Shanghai Xi Bao company), instead
Answer 24 hours, be purified through preparation liquid phase, obtain product.By the structure of mass spectral characteristi product, such as Fig. 1.
Embodiment 2-9
By embodiment 1 method, except for the difference that use oligomerization arginine and different phosphatidyl second that different Amino End Group acetic acid blocks
Hydramine reacts, and prepares the PHOSPHATIDYL ETHANOLAMINE that various oligomerization arginine is modified, such as table 1.
The PHOSPHATIDYL ETHANOLAMINE that oligomerization arginine prepared by table 1 present invention is modified
Note: in An, n represents the arginic unit number of oligomerization.
Embodiment 10
The preparation of the phosphatidyl glycerol DMPG-A4 that oligomerization arginine is modified:
The DMSO solution of 1mol A4, addition DCC (0.4g), DMAP (DMAP) (0.4g), 25 DEG C,
Priming reaction 1 hour, adds phosphatidyl glycerol 1mol, continues reaction 24 hours, is then passed through preparing liquid phase and carries out
Purifies and separates, obtains product DMPG-A4.
Embodiment 11-17
By embodiment 10 method, except for the difference that use different oligomerization arginine to react from different phosphatidyl glycerols, prepare each
Plant the phosphatidyl glycerol that oligomerization arginine is modified, such as table 2.
The phosphatidyl glycerol that oligomerization arginine prepared by table 2 present invention is modified
Embodiment 18
The preparation of the phosphatidic acid that oligomerization arginine is modified: DCC (0.4g), NHS (0.4g) join the DMSO solution of DPPA,
25 DEG C, reacted for 1 time, add A4 and react 24 hours, then pass through preparation liquid phase separation purification and obtain product DPPA-A4.
Embodiment 19-22
By embodiment 18 method, except for the difference that use different oligomerization arginine to react from different phosphatidic acid, prepare various widow
The phosphatidic acid that poly arginine is modified, such as table 3.
The phosphatidic acid that oligomerization arginine prepared by table 3 present invention is modified
Embodiment 23
The preparation of the phosphatidylcholine that oligomerization arginine is modified: p-nitrophenyl chloro-formate 0.2g, joins the DMPC of 1mol
In DMSO solution, 25 DEG C, react 12 hours, be subsequently adding A4 and react 24 hours, then pass through preparation liquid phase separation pure
Change and obtain product DMPC-A4.
Embodiment 24-27
By embodiment 23 method, except for the difference that use different oligomerization arginine to react from different phosphatidylcholines, prepare each
Plant the phosphatidylcholine that oligomerization arginine is modified, such as table 4.
The phosphatidylcholine that oligomerization arginine prepared by table 4 present invention is modified
Embodiment 28
The preparation of phospholipid nanoparticle that oligomerization arginine is modified: use ultrasonic dispersion, water 2mL, with DSPE-A4 (65mg),
DOPC (13mg), FITC-DSPE (10mg), DSPE-PEG (6mg), that cholesterol (6mg) is codissolved in 10mL methanol is molten
In agent, stirring evaporating organic solvent, residual liquid is through ultrasonic Treatment, and then hyperfiltration process isolates lipid nanoparticle, then suspendible
In aqueous solution, make lipid nanoparticle suspension.Fig. 1 and table 5 are shown in particle diameter and distribution.
Embodiment 29-51
By embodiment 28 method, first by molten in water for the water soluble drugs such as gene, polypeptide, antibody, join lipid, cholesterol
Deng methanol solvate in；Water-insoluble drug is then directly dissolved in solvent together with various lipids, prepares by embodiment 28 method
Drug-carrying nanometer particle.Such as table 5.
Can also use other liposome, the preparation method of lipid nanoparticle prepares the phospholipid nanoparticle that oligomerization arginine is modified, such as height
Press newborn even method, the emulsifying sedimentation method, microemulsion method etc..
Medicine carrying prepared by table 5 present invention or the lipid nanoparticle that blank oligomerization arginine is modified
It is 1000,2000,3000 and 5000 that PEG1, PEG2, PEG3 and PEG5 represent the molecular weight of PEG respectively；DNA used is cy5-siRNA
The structure of several lipid moleculars in table 6 table 5
Table 7 lipid nanoparticle and the fusion faculty of Hela cell membrane
| Nanoparticle | Cell membrane fusion ability/% | Nanoparticle | Cell membrane fusion ability/% |
| Embodiment 28 (NP-28) | 27 | Embodiment 45 (NP-45) | 24 |
| Embodiment 32 (NP-32) | 10 | Embodiment 46 (NP-46) | 24 |
| Embodiment 33 (NP-33) | 21 | Embodiment 47 (NP-47) | 25 |
| Embodiment 38 (NP-38) | 23 | Matched group * | 2 |
* matched group is that the DSPE-A4 in embodiment 28 is changed into DSPE.
Table 7 result shows, the nanoparticle that the phospholipid that oligomerization arginine is modified is assembled has stronger cell membrane fusion ability, permissible
Promote that nanoparticle enters intracellular efficiency, therefore, there is the ability promoting that Intracellular drug delivers effect.Transfection effect in Fig. 5
Fruit also demonstrates this point.
Characterizing method:
1. the mensuration of nanoparticle particle diameter distribution
Take 1mL nanoparticle dispersion liquid, measure its particle diameter and particle diameter distribution as laser particle analyzer.Condition determination: 25 DEG C, balance
Time 120s.LASER Light Source: He-Ne laser, wavelength 633nm.Blank nanoparticle particle diameter is distributed as in figure 2 it is shown, medicine carrying is received
Grain of rice particle diameter is distributed as shown in Figure 3.
2. the sign of lipid nanoparticle pattern
Being placed on by copper mesh in the surface plate being covered with filter paper, take about 20uL nanoparticle solution and drip on copper mesh, under room temperature, volatilization removes water.
After sample air-dries, with its pattern of transmission electron microscope observation.Electromicroscopic photograph is as shown in Figure 3.
3. DNA loads gel electrophoresis
1% agarose is joined (containing 0.5 after 1 μ L NP-A4/siRNA complex solution and 4 μ L sample-loading buffers being sufficiently mixed
μ g/mL ethidium bromide) in gel.Electrophoretic buffer is 1 × TAE, and voltage is 120V, and electrophoresis time is 40min, then exists
Observe DNA electrophoresis pattern under ultraviolet and take a picture, such as Fig. 4.
4. the sign of lipid nanometer granulosa fusion faculty
FITC fluorescence labeled fatty acid body (adding the FITC-DSPE of 10% on the basis of original lipid nanoparticle proportioning) is marked with DiI
After the cell of note cell membrane hatches 4 hours jointly, discarding culture fluid, PBS washes three times, confocal microscopy, adds up carrier
With cell membrane fluorescence overlapping cases as the index of film fusion degree.It is shown in Table 7.
5. in-vitro transfection characterizes
By 2 × 10 in 6 orifice plates5The density inoculation HeLa cell of individual cells/well, to transfection, cell culture processes is described above.
During transfection, first being changed to subtract blood serum medium by culture medium, then every hole adds the nanoparticle/DNA containing 200pmol p-DNA
The complex mass ratio 10:1 of DNA (nanoparticle with), hatches culture medium sucking-off in orifice plate after 4h, with 1 in cell culture incubator
ML PBS washes three times with removing not by the nano-complex of endocytosis, then will add 1mL after cell dissociation with 0.2mL pancreatin
In DMEM and pancreatin, at 4 DEG C, 1000rpm rotating speed is centrifuged 8min, then after washing three times with 0.8mL PBS, by cell with 0.3
ML PBS disperses, and with flow cytomery green fluorescent protein positive cell, INSTRUMENT MODEL is BD FACSVerse.Result
See Fig. 5.
Claims (10)
1. the phospholipid that an oligomerization arginine is modified, it is characterised in that described oligomerization arginine is containing 4-8
The oligomerization arginine of arginine unit, modifies on the phosphoric acid head base of phospholipid.
2. the phospholipid that oligomerization arginine as claimed in claim 1 is modified, is characterized in that described phospholipid is phosphorus
Acyl ethanolamine, phosphatidylcholine, phosphatidyl glycerol, phosphatidic acid, Phosphatidylserine or phospholipid
Acyl inositol.
3. the phospholipid that oligomerization arginine as claimed in claim 2 is modified, is characterized in that the carbon of described phospholipid
Hydrogen chain contains 12-20 carbon atom.
4. the phospholipid that the oligomerization arginine as described in claim 1-3 is modified, is characterized in that described oligomerization essence
Propylhomoserin preferably comprises the oligomerization arginine of 4 or 5 arginine unit, and described phospholipid at least contains
2 hydrocarbon chains.
5. the phospholipid that oligomerization arginine as claimed in claim 4 is modified, is characterized in that described phospholipid is selected from
Dilauroyl PHOSPHATIDYL ETHANOLAMINE, two myristoyl PHOSPHATIDYL ETHANOLAMINE, two palmityl phosphorus
Acyl ethanolamine, DSPE, two Semen arachidis hypogaeae acylphosphatidyl ethanolamines, two
Oleolyl phosphatidyl ethanolamine, GLYCEROL,DIMYRISTOYL PHOSPHATIDYL, DSPG,
DLPG, DPPG, DOPG, egg
The acid of yellow phosphorus phosphatidyl glycerol, dioleoyl phospholipid, two myristoyl phospholipid, DPPA, two
Stearyl phosphatidic acid, dilauroyl phosphatidic acid, dilauroyl phosphatidic acid, two myristoyl
Phosphatidylcholine, Dioctonoyl pnosphotidyl choline, DSPC, two Semen arachidis hypogaeae acyls
Base phosphatidylcholine or dioleyl phosphatidyl choline.
6. the preparation method of the phospholipid that the oligomerization arginine described in claim 1-5 is modified, is characterized in that passing through
Prepared by following method: oligomerization arginine peptide chain dicyclohexylcarbodiimide/N-hydroxysuccinimidyl acyl
After imines activation, react with PHOSPHATIDYL ETHANOLAMINE, obtain, through purification, the phosphorus that oligomerization arginine is modified
Acyl ethanolamine；Or oligomerization arginine peptide chain dicyclohexylcarbodiimide/4-dimethylamino pyrrole
After pyridine activation, react with phosphatidyl glycerol, obtain, through purification, the phosphatidyl that oligomerization arginine is modified
Glycerol；Or oligomerization arginine peptide chain is lived with dicyclohexylcarbodiimide/N-hydroxy-succinamide
The phosphatidic acid reaction changed, obtains, through purification, the phosphatidic acid that oligomerization arginine is modified；Or oligomerization essence
Propylhomoserin peptide chain reacts with the phosphatidylcholine of p-nitrophenyl chloroformate activation, obtains through purification
The phosphatidylcholine that oligomerization arginine is modified.
7. the nanoparticle that the phospholipid that the oligomerization arginine described in claim 1-5 is modified is assembled, is characterized in that
The phospholipid that oligomerization arginine is modified assembles jointly with other lipid, forms lipid nanoparticle or liposome,
Particle diameter is 30～200nm, wherein oligomerization arginine modify phospholipid in nanoparticle quality hundred
Dividing content is 5%～80%.
8. the nanoparticle that the phospholipid that oligomerization arginine as claimed in claim 7 is modified is assembled, is characterized in that
Phospholipid, phosphatidylcholine, cholesterol and the Polyethylene Glycol modified containing oligomerization arginine in nanoparticle
The PHOSPHATIDYL ETHANOLAMINE modified, the phospholipid mass content that wherein oligomerization arginine is modified is 10%～80%,
Cholesterol mass content 5%～50%, the mass content of phosphatidylcholine are 1%～50%, poly-second two
The PHOSPHATIDYL ETHANOLAMINE mass content that alcohol is modified is 0.5%～10%.
9. the nanoparticle that the phospholipid that oligomerization arginine as claimed in claim 8 is modified is assembled, is characterized in that
The phospholipid that oligomerization arginine containing 4 or 5 arginine unit is modified and cholesterol, DOPC or
DSPC, polyethyleneglycol modified DSPE (DSPE-PEG) assemble and are formed
Nanoparticle, wherein, the molecular weight of PEG preferably 1000～5000.
10. the application of the nanoparticle that the phospholipid that the oligomerization arginine described in claim 7-9 is modified is assembled, its
It is characterised by loading gene, siRNA, polypeptide or protein medicaments, antibody and chemistry
Medicine, the aqueous dispersions of the drug-carrying nanometer particle of formation, for cell in vitro or internal local, vein
Injected delivery medicine.
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| CN114007653A (en) * | 2019-05-05 | 2022-02-01 | 复旦大学 | Drug delivery vehicle and pharmaceutical preparation using the same |
| CN114007653B (en) * | 2019-05-05 | 2024-02-09 | 复旦大学 | Drug delivery vehicle and pharmaceutical formulation using the same |
| CN111944851A (en) * | 2019-05-16 | 2020-11-17 | 天津大学 | Nanoscale siRNA delivery system |
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| CN110404080B (en) * | 2019-07-09 | 2021-06-22 | 南京大学 | Mitochondrion targeted phospholipid and application thereof |
| CN112043670A (en) * | 2019-10-08 | 2020-12-08 | 中国药科大学 | Mitochondria-targeted external medicinal preparation |
| CN112043670B (en) * | 2019-10-08 | 2021-10-29 | 中国药科大学 | Mitochondria-targeted external medicinal preparation |
| WO2022013306A1 (en) * | 2020-07-14 | 2022-01-20 | Universität Heidelberg | Oral pharmaceutical compositions comprising lipid conjugates |
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