CN107488137A - A kind of quaternary ammonium salt of small-molecular-weight and its preparation method and application - Google Patents
A kind of quaternary ammonium salt of small-molecular-weight and its preparation method and application Download PDFInfo
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Abstract
The present invention relates to a kind of quaternary ammonium salt of small-molecular-weight, the cationic structural formula of the quaternary ammonium salt is as follows：Wherein, R1And R2Independently be expressed as C2~C4Alkyl, R3Independent is expressed as amino, hydroxyl, carboxyl, aldehyde radical, sulfydryl, diazanyl or succinimido.The invention further relates to its preparation method and application, contain disulfide bond in the cationic structural of the quaternary ammonium salt of the small-molecular-weight, can decompose in the cell in the presence of reduced glutathione, reduce its cytotoxicity, improve the release of nucleic acid drug.
Description
Technical field
The present invention relates to the compound for carrying nucleic acid drug, and in particular to a kind of quaternary ammonium salt of small-molecular-weight and its
Preparation method and application.
Background technology
Gene therapy has broad application prospects in human body gene defect or abnormal caused disease treatment, but by
In nucleic acid drug in vivo it is unstable, be easily degraded and be difficult to be absorbed into target cell so that the delivering skill of nucleic acid drug
Art turns into research and development focus in the last few years.
Compared to viral vector, non-virus carrier has the advantages that to be readily synthesized, low immunogenicity and security are higher, tool
There is preferable application prospect, it mainly includes cation lipid and the major class of cationic polymer two, such as cationic-liposome
Lipofectamine 2000, cationic polymer polyethyleneimine (PEI), polylysine, polyglutamic acid, polyamidoamine
Tree, polyethyl methacrylate and some natural polymers such as chitosan etc..Mostly in life in these non-virus carriers
Lotus positive electricity under environment is managed, is combined available for absorption or compresses electronegative nucleic acid drug, protect it from the degraded of nuclease.
But the efficiency gene transfection of current non-virus carrier is still relatively low on the whole, except the band due to its electropositive
Easily be eliminated inside coming and nonspecific cellular uptake (lacking targeting) outside, what itself and nucleic acid drug were formed
Compound has higher stability, causes nucleic acid drug to be difficult to discharge in the cell, is to influence its efficiency gene transfection
One key factor.
Therefore, the carrier that design is easy to dissociate is more suitable for the release of nucleic acid drug, and existing a variety of biostimulations ring at present
The report of the non-virus carrier of answering property release, its biological trigger mechanism is including pH, temperature, oxidation or reducing environment and specifically
Property expression of enzymes etc..But the most of carrier material studied at present, after response fracture occurs, caused cationic materials or piece
Duan Yiran can produce stronger absorption with nucleic acid drug.Research is found in addition, and cationic polymer has obvious cell
Toxicity, the toxicity have Structure Dependence, molecular weight, charge density, steric configuration and cation with cationic polymer
The factors such as the surface potential of polymer-DNA compounds are relevant.For the similar cationic polymer of structure, molecular weight and carrier
The density of surface charge is to influence the key factor of carrier toxicity, and molecular weight is higher, and the charge density of carrier surface is bigger, carrier
Toxicity it is bigger.
It would therefore be highly desirable to synthesize a kind of little molecular cation peptides bioactivator quaternary ammonium salt with biostimulation response, it is incorporated into tool
Have in the macromolecular of good biocompatibility, for carrying nucleic acid drug, and disadvantages mentioned above can be overcome.
The content of the invention
In view of the above-mentioned deficiencies in the prior art, it is an object of the present invention to provide quaternary ammonium salt and its preparation side of a kind of small-molecular-weight
Method and application, contain disulfide bond in the cationic structural of the quaternary ammonium salt of the small-molecular-weight, can be in the work of reductive glutathione
With lower fracture, improve the release of nucleic acid drug；Secondly as its molecular weight is smaller, the charge density on surface is suitable, reduces it
Cytotoxicity.
The present invention is directed to the technical scheme that above-mentioned technical problem is provided：
A kind of quaternary ammonium salt of small-molecular-weight, the cationic structural formula of the quaternary ammonium salt are as follows：
Wherein, R1And R2Separately it is expressed as C2~C4Alkyl, R3Independently it is expressed as amino, hydroxyl, carboxyl, aldehyde
Base, sulfydryl, diazanyl or succinimido.
Because above-mentioned quaternary ammonium salt molecular weight is smaller, less cytotoxicity is made it have；Secondly, in cationic structural
N, N, N- trimethyl have positive charge, can adsorb negatively charged nucleic acid drug, and the R of the cationic structural formula other end3
For the group available for engrafted nanometer carrier, nano-carrier can be the endogenous polysaccharide with good biocompatibility, such as
Hyaluronic acid etc. so that above-mentioned quaternary ammonium salt can be used for carrying nucleic acid drug after suitable nano-carrier is modified.
In addition, containing disulfide bond in the cationic structural of quaternary ammonium salt, can be broken in the presence of reductive glutathione, paddy
The natural activity peptide that the sweet peptide of Guang is made up of glutamic acid, cysteine and glycine, wherein reduced glutathione (GSH) is thin
The concentration (about 0.5~10mM) in intracellular portion is more than 100 times of concentration in extracellular physiological environment (2~20 μM), in the present invention
Quaternary ammonium salt after nucleic acid drug is carried, can in the cell reproducibility response release nucleic acid drug, improve nucleic acid
The release of medicine.
Preferably, the anion of the quaternary ammonium salt is acid ion.
Preferably, the structural formula of the quaternary ammonium salt is as follows：
The present invention also provides a kind of preparation method of the quaternary ammonium salt of small-molecular-weight described above, comprises the following steps：
1) 2-aminoethyl disulfide dihydrochloride carries out BOC protection reactions with di-tert-butyl dicarbonate, obtains the cystamine of one end BOC protections；
2) cystamine of one end BOC protections carries out substitution reaction with KI, obtains the N of one end BOC protections, N, N- trimethyls
Cystamine salt compounded of iodine；
3) N of one end BOC protections, N, N- trimethyl cystamine salt compounded of iodine carry out de- BOC reactions with trifluoroacetic acid, obtain N, N, N-
Trimethyl cystamine trifluoroacetate.
Preferably, BOC protections reaction is specially in the step 1)：Di-tert-butyl dicarbonate solution is added to containing three second
Reacted in the 2-aminoethyl disulfide dihydrochloride solution of amine, after removing solvent, add sodium dihydrogen phosphate, removing two is washed with ether
The cystamine of BOC protections is held, then adjusts pH to alkalescence, the cystamine for one end BOC protections being obtained by extraction.
Preferably, the mass ratio of di-tert-butyl dicarbonate and 2-aminoethyl disulfide dihydrochloride is 0.8~1.2 in the step 1):1.
Preferably, the rate of charge of 2-aminoethyl disulfide dihydrochloride and triethylamine is 200mg in the step 1):350~400 μ l.
Preferably, the molar concentration of sodium dihydrogen phosphate is 0.8~1.2M in the step 1), more preferably
1M。
Preferably, regulation pH to 8~10, more preferably pH=9 in the step 1).
Preferably, substitution reaction is specially in the step 2)：K is added in the solution of the cystamine of one end BOC protections2CO3With
Iodomethane carries out lucifuge reaction, removes K2CO3And solvent, obtain the N of one end BOC protections, N, N- trimethyl cystamine salt compounded of iodine.
Preferably, the cystamine and the mol ratio of iodomethane that one end BOC is protected in the step 2) are 1:3~6, it is further excellent
Elect 1 as:5.
Preferably, the cystamine and K that one end BOC is protected in the step 2)2CO3Mass ratio be 1:1~1.4.
Preferably, the reaction temperature that lucifuge is reacted in the step 2) is room temperature, and the reaction time is 2~4 days, further excellent
Elect as 3 days.
Preferably, BOC reactions are taken off in the step 3) is specially：The N of one end BOC protections, N, N- trimethyl cystamine salt compounded of iodine
Trifluoroacetic acid reaction is added in solution, product is obtained after removing solvent, continues to dissolve product, it is residual for neutralizing to add triethylamine
The trifluoroacetic acid stayed, N, N, N- trimethyl cystamine trifluoroacetates are obtained after removing solvent.
Preferably, mole of the N that one end BOC is protected in the step 3), N, N- trimethyl cystamine salt compounded of iodine and trifluoroacetic acid
Than for 1:12~16, more preferably 1:14.3.
The present invention also provides a kind of application of quaternary ammonium salt of small-molecular-weight described above in nucleic acid drug is carried.
Preferably, the nucleic acid drug includes oligonucleotides, DNA, siRNA or microRNA.Further preferably
For siTwist, Twist is a kind of highly conserved dHAND gene family important member, overexpression
CAM 120/80 function can be suppressed, cause cell-cell adhesion action deprivation so that cell has invasive ability, therefore Twist is swollen
It played an important role in knurl invasion and attack transfer.Twist genes can be overcome the multidrug resistance of tumour as new target spot
Property.
Compared with the existing technology, beneficial effects of the present invention are embodied in：
(1) the quaternary ammonium salt molecular weight in the present invention is smaller, has less toxicity.
(2) N in quaternary ammonium salt its cationic structural in the present invention, N, N- trimethyls have positive charge, can adsorb band
The nucleic acid drug of negative electrical charge so that above-mentioned quaternary ammonium salt can be used for carrying nucleic acid medicine after suitable nano-carrier is modified
Thing.
(3) disulfide bond is contained in quaternary ammonium salt its cationic structural in the present invention, can be in the work of reductive glutathione
With lower fracture, improve the release of nucleic acid drug.
Brief description of the drawings
Fig. 1 is cystamine, cystamine-BOC, N, N in embodiment 1, N- trimethyl cystamine salt compounded of iodine-BOC and N, N, N- trimethyl Guang
The nuclear magnetic spectrogram of amine trifluoroacetate；
Fig. 2 be embodiment 1 in N, N, the carbon spectrogram of N- trimethyl cystamine trifluoroacetates；
Fig. 3 be embodiment 3 in N, N, N- trimethyl cystamine salt compounded of iodine-BOC mass spectrogram；
Fig. 4 be embodiment 3 in N, N, the mass spectrogram of N- trimethyl cystamine chlorates；
Fig. 5 is the agarose gel electrophoresis figure of control sample and different quality than the HSTC/siRNA of mixing in application examples 2；
Fig. 6 is that control sample and different quality coagulate than the HSTC/siRNA of mixing agarose after DTT is handled in application examples 2
Gel electrophoresis figure；
Fig. 7 is release fluorograms of the siRNA in cell in application examples 3.
Embodiment
With reference to specific embodiment, the present invention is further illustrated.
Embodiment 1:Synthesize N, N, N- trimethyl cystamine trifluoroacetates
1) 200mg 2-aminoethyl disulfide dihydrochlorides (being designated as cystamine) are weighed to be dissolved in 10ml methanol, and add 386 μ l triethylamines；
192.5mg di-tert-butyl dicarbonates (BOC acid anhydrides) are dissolved in 2ml methanol, are added dropwise to 2-aminoethyl disulfide dihydrochloride methanol solution
In, reaction 30min is stirred at room temperature, is evaporated methanol.Continuously add 50ml sodium dihydrogen phosphate (NaH2PO4, 1M), use
Ether is washed twice to remove the product that both ends all connect upper BOC acid anhydrides, the pH to 9 using NaOH (1M) with regulation solution, uses second
The cystamine of one end BOC protections is obtained by extraction in acetoacetic ester, is evaporated ethyl acetate layer, obtain white product (be designated as cystamine-
BOC)。
2) the above-mentioned products of 50mg are weighed, are dissolved in 1ml acetonitriles, add 64.6mg K2CO3With 73 μ l iodomethane, room temperature is kept away
Light reaction 3d, centrifugation remove K2CO3, supernatant is evaporated, is washed 2 times with ether, white product is obtained and (is designated as N, N, N- tri-
Methyl cystamine salt compounded of iodine-BOC).
3) above-mentioned white product is dissolved with 1ml methanol, adds 1ml trifluoroacetic acids, reacts 2h, now solution becomes yellowish-brown
Color, 2ml methanol solutions are dissolved in after being evaporated again, add the trifluoroacetic acid in 30 μ l triethylamines and remained, be evaporated methanol
Obtain N, N, N- trimethyl cystamine trifluoroacetates.
For the cystamine in embodiment 1, cystamine-BOC, N, N, N- trimethyl cystamine salt compounded of iodine-BOC and N, N, N- trimethyl Guang
Amine trifluoroacetate, uses D2O dissolves as solvent, carries out1H-NMR is analyzed, as shown in Figure 1.
As a result show：The characteristic peak of cystamine methylene BOC institutes band in 2.9ppm and 3.3ppm, intermediate product cystamine-BOC
The characteristic peak for three methyl having is in 1.3ppm, due to the effect of carbonyl in BOC so that in cystamine on the methylene of carbonyl
The characteristic peak of hydrogen be subjected to displacement 2.7ppm to the right.The product N, N, N- tri- of three methyl is further modified on cystamine-BOC
Methyl cystamine salt compounded of iodine-BOC, the characteristic peak of its three methyl is in 3.1ppm.After de- BOC protections, three methyl that BOC is carried
Characteristic peak disappears, it was demonstrated that the successful synthesis of N, N, N- trimethyl cystamine trifluoroacetate.
For the N in embodiment 1, N, N- trimethyl cystamine trifluoroacetates, carbon spectrum analysis is carried out, as shown in Fig. 2 its
In, the chemical shift of 1 chemical shift, 33.3,2 chemical shift 37.74,3 is the chemical shift of 30.02,4 chemical shift 65.33,5
53.21, the results showed that the successful synthesis of N, N, N- trimethyl cystamine trifluoroacetate.
Embodiment 2:Synthesize N, N, N- trimethyl cystamine iodized salts
(1) 500mg 2-aminoethyl disulfide dihydrochlorides (being designated as cystamine) are weighed to be dissolved in 10ml methanol, and add 965 μ l triethylamines；
Reaction 30 minutes, 223mg benzyl chloroformates (CBZ) are dissolved in 2ml methanol, are added dropwise to 2-aminoethyl disulfide dihydrochloride methanol solution
In, reaction is stirred at room temperature, by thin-layered chromatography observing response progress, after raw material point disappearance, is evaporated methanol.Continue to add
Enter hydrochloric acid solution (HCl, 1M), washed with ether twice to remove the product that both ends all connect upper CBZ acid anhydrides, used using NaOH (1M)
The pH to 9 of solution is adjusted, is extracted with ethyl acetate to obtain the cystamine of one end CBZ protections, is evaporated ethyl acetate layer, obtains white
The product (being designated as cystamine-CBZ) of color.
(2) cystamine-CBZ 39mg are weighed, add 64.6mg K2CO3, 73 μ L iodomethane are dissolved in 1ml acetonitriles, react 3d,
Centrifugation removes K2CO3, supernatant is evaporated, is washed 2 times with ether, obtains white product, adds dichloromethane and petroleum ether (volume
Than 3:1) washing rotation is evaporated, and repeated washing is spin-dried for 2 times, removes acetonitrile.
(3) methanol (3ml) lysate is used, adds 15%Pd/C (0.026g), is hydrogenated 12 hours at room temperature, obtains N,
N, N- trimethyl cystamine iodized salt.
Embodiment 3:Synthesize N, N, N- trimethyl cystamine chlorates
The present embodiment adds 1ml hydrochloric acid saturation ethyl acetate (3M) when removing de- BOC, reacts at room temperature 30 minutes, is spin-dried for solvent,
Remaining step is same as Example 1, is prepared into solid N, N, N- trimethyl cystamine chlorate.
As a result show：For the N in embodiment 3, N, N- trimethyl cystamine salt compounded of iodine-BOC and N, N, N- trimethyl cystamine chlorine
Salt dissolving carries out mass spectral characteristi, as shown in figs. 34.N, N, N- trimethyl cystamine salt compounded of iodine-BOC molecular ion peak ([M+H]+) be
295.17, and the molecular ion peak ([M+H] of N, N, N- trimethyl cystamine chlorate+) it is 195.10, thus prove N, N, N- tri-
Methyl cystamine chlorate successfully synthesizes.
Application examples 1：Synthesize HSTC/siRNA
(1) 100mg hyaluronic acids (HA, Mw 48kDa) are weighed to be dissolved in 30ml deionized waters, 50mg is added dropwise
NaIO4The aqueous solution, 2d is reacted under the conditions of room temperature lucifuge, add 0.2ml ethylene glycol, stir terminating reaction after 1h.After collecting product
With 3500WM bag filters dialysis 3d, dialysis medium is deionized water, is placed in 4 DEG C of preservations after lyophilized, obtains product HA-CHO.
(2) weigh the above-mentioned product HA-CHO of 62mg and 318mg spermine (spermine) be dissolved in PBS (PH7.4-7.8,
Stirring 4h, can produce some precipitations in 0.1M), add NaOH (1M) regulation pH until precipitation dissolves.Add the cyano group boron of 10 times of amounts
12h is dialysed to remove the spermine on unreacted with 3500WM bag filters after sodium hydride reaction 3d, collection product HA-spermine,
Dialysis medium is deionized water, that is, obtains HA-spermine.
(3) 72mg adipic dihydrazides (ADH) and 80mg DOX-TPP are dissolved in 15ml absolute methanols, are formed and be suspended
Liquid, nitrogen protection stirring 30min, adds 100 μ l trifluoroacetic acids, 24h is stirred under the conditions of room temperature lucifuge.It is evaporated methanol
Afterwards, ethanol lysate is added, centrifugation removes unreacted ADH, is evaporated alcohol layer again, obtains red product ADH-
DOX-TPP。
(4) 31mg HA-spermine are weighed to be dissolved in 10ml deionized waters, 30mg EDC and 18mg NHS are dissolved
In 2ml deionized waters, after 1h is reacted in regulation to pH 5.0, pH7.4 is arrived in regulation, and 4ml ADH-DOX-TPP methanol is added dropwise
Solution (10mg/ml) reacts 4h, adds the N of 1.5 times of amounts, N, N- trimethyl cystamines trifluoroacetate (prepared by embodiment 1), reaction
Overnight, with 3500WM bag filters dialysis 3d after collection product, dialysis medium is deionized water, is placed in 4 DEG C of preservations after lyophilized, produces
To N, N, N- trimethyls cystamine-hyaluronic acid-spermine-adriamycin-triphenylphosphine nanometer formulation (48kDa HSTC).
(5) it is 100 in mass ratio by nanometer formulation HSTC and siRNA:1 is mixed in DEPC water, 37 DEG C, 100rpm constant temperature
30min is shaken, obtains N, N, N- trimethyls cystamine-hyaluronic acid-spermine-adriamycin-triphenylphosphine+siRNA nanometer formulations
(HSTC/siRNA)。
Application examples 2：48kDa HSTC combinations siRNA efficiency and glutathione response siRNA release is investigated
48kDa HSTC judge that siRNA binding ability step is as follows by agarose gel electrophoresis experiment：
The HSTC preparations of 1 μ l siRNA solution (100ng/ μ l) and different quality (0.5 μ g, 2.5 μ g, 5 μ g, 10 μ g, 15 μ g) are carried out
The solution that cumulative volume is 21 μ l is mixedly configured into, the DEPC water of the addition respective volume less than 21 μ l is supplied, and is mixed uniformly,
After placing 37 DEG C of constant temperature oscillation case vibration 30min, by different quality ratio ((5:1、25:1、50:1、100:1、150:1)) mix
HSTC/siRNA, 4 μ l Loading buffer are added, be well mixed.Configure the agar that the concentration containing 0.1%GelRed is 2%
Sugared gel, after carrying out 30min electrophoresis under the conditions of voltage 100V, place and observed under uviol lamp.
As a result show, be grafted N on HSTC, N, N- trimethyl cystamines carry positive charge in neutral conditions, in siRNA
Property under the conditions of carry negative electrical charge, both are combined by the electrostatic interaction of positive and negative charge.Agarose gel electrophoresis can be intuitively anti-
Reflect HSTC and siRNA joint efficiency.Under certain deposition condition, the siRNA that dissociates is moved in the presence of electric field force to positive pole.
And after forming compound with HSTC, siRNA is arrested in well.By HA-spermine and HSTC with different mass ratioes with
SiRNA is incubated 30min under the conditions of 37 DEG C, and PEI is as positive control, as a result as shown in figure 5, with HSTC and siRNA mass
The increase of ratio, the fluorescence intensity of free siRNA bands gradually weaken.
To investigate the release of HSTC/siRNA external GSH responses, by HSTC/siRNA of the different quality than mixing, with two
After sulphur threitol (DTT, 50mM or 200mM) processing 2h, subsequently ibid Operations Analyst.
As shown in fig. 6, after being handled with 200mM DTT, it is observed that siRNA dissociates again shows band.Quality
Than under 25 and 50, siRNA discharges completely, mass ratio 100, about 50% Gene releaser, mass ratio continues to increase to 150, only
There is more at least part of Gene releaser.When DTT concentration is 50mM, the identical time is handled, siRNA release is reduced.And PEI
Group, siRNA is not discharged then after DTT processing.
Conclusion：HSTC is to 50 with siRNA mass ratioes:When 1, most of siRNA is combined with HSTC.When HSTC with
SiRNA mass ratio is 100:When 1, both are completely combined.And intermediate product HA-spermine can not under all mass ratioes
With reference to siRNA.Carrier strengthens with the rise of carrier and gene mass ratio the binding ability of gene, and DTT destroys disulfide bond
SiRNA is caused to discharge fewer.When DTT concentration rise, more, base of siRNA release stronger to the destructive power of disulfide bond
Because carrier has the function of the sensitive control siRNA of reproducibility release.
Application examples 3：SiRNA intracellular release experiment
By the MCF-7/ADR cells of exponential phase with every hole 2 × 104Individual cell is inoculated in chamber cover glass, 37
DEG C, 5%CO2Under the conditions of cultivate 24h after, remove nutrient solution, add ethyl ester containing glutathione reduction (GSH-Oet, concentration are
Nutrient solution processing cell 2h 20mM), adds 48kDa HSTC/FAM-siRNA (concentration 100nM), is positioned over cell culture
After case is incubated 2h, using PBS 3 times, the PBS that 100 μ l contain 10% hyclone is added, uses confocal microscopy
Release conditions of the FAM-siRNA in MCF-7/ADR cells, as shown in Figure 7.
Conclusion：Show rose (as shown in white arrow 1) and lavender fluorescence (such as white in blank control group image
Shown in arrow 2), green fluorescence is not shown, illustrates that siRNA is also combined by HSTC and does not discharge, and GSH-OEt treatment groups,
Into the cell it is observed that green fluorescence (as shown in white arrow 3), illustrates that GSH is horizontal and improve the release that can accelerate siRNA.
Claims (9)
1. a kind of quaternary ammonium salt of small-molecular-weight, it is characterised in that the cationic structural formula of the quaternary ammonium salt is as follows：
Wherein, R1And R2Separately it is expressed as C2~C4Alkyl, R3Independently be expressed as amino, hydroxyl, carboxyl, aldehyde radical,
Sulfydryl, diazanyl or succinimido.
2. the quaternary ammonium salt of small-molecular-weight according to claim 1, it is characterised in that the anion of the quaternary ammonium salt is acid group
Ion.
3. the quaternary ammonium salt of small-molecular-weight according to claim 1, it is characterised in that the structural formula of the quaternary ammonium salt is as follows：
4. a kind of preparation method of the quaternary ammonium salt of small-molecular-weight as claimed in claim 3, it is characterised in that including following step
Suddenly：
1) 2-aminoethyl disulfide dihydrochloride carries out BOC protection reactions with di-tert-butyl dicarbonate, obtains the cystamine of one end BOC protections；
2) cystamine of one end BOC protections carries out substitution reaction with KI, obtains the N of one end BOC protections, N, N- trimethyl cystamines
Salt compounded of iodine；
3) N of one end BOC protections, N, N- trimethyl cystamine salt compounded of iodine carry out de- BOC reactions with trifluoroacetic acid, obtain N, N, N- front threes
Base cystamine trifluoroacetate.
5. the preparation method of the quaternary ammonium salt of small-molecular-weight according to claim 4, it is characterised in that in the step 1)
BOC protection reaction be specially：Di-tert-butyl dicarbonate solution is added in the 2-aminoethyl disulfide dihydrochloride solution containing triethylamine and carried out
Reaction, after removing solvent, sodium dihydrogen phosphate is added, the cystamine for removing both ends BOC protections is washed with ether, then adjusts pH
To alkalescence, the cystamine for one end BOC protections being obtained by extraction.
6. the preparation method of the quaternary ammonium salt of small-molecular-weight according to claim 4, it is characterised in that taken in the step 2)
In generation, reacts：K is added in the solution of the cystamine of one end BOC protections2CO3Lucifuge reaction is carried out with iodomethane, removes K2CO3
And solvent, obtain the N of one end BOC protections, N, N- trimethyl cystamine salt compounded of iodine.
7. the preparation method of the quaternary ammonium salt of small-molecular-weight according to claim 4, it is characterised in that taken off in the step 3)
BOC reacts：N, the N of one end BOC protections, trifluoroacetic acid reaction is added in N- trimethyl cystamine iodized salt solutions, is removed molten
Product is obtained after agent, continues to dissolve product, the trifluoroacetic acid that triethylamine is used to neutralize residual is added, N is obtained after removing solvent,
N, N- trimethyl cystamine trifluoroacetate.
A kind of 8. application of the quaternary ammonium salt of small-molecular-weight as described in claims 1 to 3 is any in nucleic acid drug is carried.
9. application of the quaternary ammonium salt of small-molecular-weight according to claim 8 in nucleic acid drug is carried, it is characterised in that
The nucleic acid drug includes oligonucleotides, DNA, siRNA or microRNA.
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| CN105143456A (en) * | 2013-03-15 | 2015-12-09 | 不列颠哥伦比亚大学 | Lipid nanoparticles for transfection and related methods |
| WO2017081100A1 (en) * | 2015-11-09 | 2017-05-18 | Shell Internationale Research Maatschappij B.V. | Process for removing mercaptans from a gas stream |
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| CN105143456A (en) * | 2013-03-15 | 2015-12-09 | 不列颠哥伦比亚大学 | Lipid nanoparticles for transfection and related methods |
| WO2017081100A1 (en) * | 2015-11-09 | 2017-05-18 | Shell Internationale Research Maatschappij B.V. | Process for removing mercaptans from a gas stream |
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| CN111410757A (en) * | 2020-03-31 | 2020-07-14 | 江南大学 | Preparation method of degradable and environment-responsive compound microgel |
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