CN108619098A - A kind of Raltitrexed pH sensitive liposomes and preparation method - Google Patents
A kind of Raltitrexed pH sensitive liposomes and preparation method Download PDFInfo
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Abstract
The invention discloses a kind of Raltitrexed pH sensitive liposomes and preparation methods.The first gained encapsulation rate of Raltitrexed pH sensitive liposomes prepared by the present invention is between 45.75%~49.95%, and final gained encapsulation rate is up to 89.45%~95.23%；Quality is the suspension liquid of the general blue-opalescent of white, and uniform particle diameter favorable dispersibility；Zeta potential is between 46.6mv~42.5mv, excellent in stability；Grain size is suitable as the carrier of administration in 217nm~278nm；In vitro in release experiment, gained liposome is quick in acidic environment (pH5.0~6.5) release of tumor tissues, and release is slow in normal structure pendular ring border (pH7.4), and pH sensitizations are apparent；Cytotoxicity experiment shows that gained liposome inhibitor against colon carcinoma cells cell effect is excellent, can be effectively targeted to tumor tissues；And ensure blank liposome nontoxicity, toxic side effect is few.
Description
Technical field
The present invention relates to the passive target fields that pH in liposome field and tumor tissues is responded, and in particular to a kind of thunder replaces
Song plug pH sensitive liposomes and preparation method.
Background technology
Raltitrexed is a kind of quinazoline folate analog, is played by inhibiting thymine synthetase (TS) anti-swollen
Tumor acts on, and antitumor action is from not only drug itself, and after drug enters body, only 40%~50% with original in 48h
Type drug is drained, and a part of drug is actively absorbed by reduced form folic acid methotrexate (MTX) cell membrane carrier by cell, by folic acid
Polyglutamic acid synzyme is metabolized to a series of polyglutamic acid class compounds, and such compound has thymidylate synthase stronger
Inhibiting effect, and can be detained for a long time in the cell, play long term toxicity.Compared with 5-FU, Raltitrexed has stronger anti-
The activity of human colon carcinoma tumour cell has 16 countries as colorectal cancer fiest-tire medication now.
The long half time of Raltitrexed reaches 198h, therefore has longer chemotherapy intermittent phase, every 3 weeks medication in use
1 time, current listing preparation both domestic and external is freeze-dried powder used for intravenous injection.Although the clinic of its freeze-dried powder used for intravenous injection is answered
With more and more extensive, but also there is apparent limitation.Since the tumour that cannot efficiently differentiate normal cell and fast breeding is thin
Born of the same parents, so that it is reached, the dose of tumor tissues is less, adverse reaction is larger, and the clinical examination of advanced colorectal cancer is treated from Raltitrexed
It can be found that the main adverse reaction of single one line of medicine Raltitrexed or second line treatment is Neuroleptic Leukocytopenia, neutrophil leucocyte in testing
Reduction, Nausea and vomiting, weak, apocleisis and glutamic-pyruvic transaminase increase, incidence is respectively 9%~13.8%, 23%, 20%,
6.9%~9%, 14% and 7%.Therefore, finding, which reduces the method that toxic side effect improves curative effect, becomes urgent problem to be solved.
Due to higher metabolic rate, the pH value of tumor microenvironment and inflammatory tissue is 5.0~6.5, intracellular different thin
The pH value of born of the same parents' device is also different, if the pH value of endosome (Endosome) is 5.5, the pH value of lysosome (Lysosome) is 5.0, this
The microenvironment of a little difference pH value provides thinking for the design of pH responsive polymer nano-particles.PH responsive type liposomes are can
Effectively in a kind of new drug carrier of specific time and privileged site release institute entrapped drug, because it is with good biology
Compatibility, targeting, slow release, reduce drug toxicity and improve drug stability the effects that and by extensive concern.
Although pH sensitive liposomes have certain report, there is no Raltitrexed is prepared as pH sensitivities for current this field
The research of liposome, when preparing pH sensitive liposomes using Raltitrexed, in order to ensure that it can be effectively targeted to tumor group
The toxicity of drug is knitted, reduced, its antitumor action is preferably played, it is necessary to it is good to ensure that Raltitrexed liposome has
Encapsulation rate, pH sensibility and stability, but it is extremely difficult to meet this three.
Chinese patent CN 2011102286230 discloses one kind<Antitumor pH sensitive liposomes, its freeze-dried powder
And their preparation method>Although the final gained encapsulation rate of its liposome prepared is between 60%~90%, it is encapsulated
Rate amplitude of variation is big, and stability is poor, it is caused to be difficult to industrialized production use.
Therefore how to provide that a kind of encapsulation rate is high, sensibility is strong, stability is good, excellent quality Raltitrexed liposome is
This field urgent problem to be solved.
Invention content
To solve the above-mentioned problems, the present invention provides a kind of encapsulation rate height, the Raltitrexeds that sensibility is strong, stability is good
Liposome.
The invention discloses a kind of Raltitrexed pH sensitive liposomes, the Raltitrexed pH sensitive liposomes, by weight
Number meter, including following components：5~15 parts of Raltitrexed, 35~105 parts of phosphatidyl-ethanolamine, cholesterol succinate 15~
45 parts.
In order to provide the Raltitrexed pH sensitive liposomes that a kind of encapsulation rate is high, sensibility is strong, stability is good, inventor tastes
It has tried a variety of membrane materials to be prepared, according to Zeng Huilin, Wang Shanshan, symbol rising sun east .pH sensitive liposomes are in drug delivery system
Using [J] medicine Leaders, 2014,33 (03):The research of 348-351. etc. is it is found that phospholipid, oleic acid, cholesterol etc. can be with
Apply in the research of liposome, however, not all membrane material can obtain excellent effect, in repeatedly grinding for inventor
Middle discovery is studied carefully, in the membrane material for preparing Raltitrexed, only using phosphatidyl-ethanolamine and cholesterol succinate as preparing material
The first gained encapsulation rate of liposome just be can guarantee between 45.75%~49.95%, final gained encapsulation rate is up to 89.45%
~95.23% effect, if according to comparative example 1 it is found that membrane material is replaced with soybean lecithin, cholesterol, the liposome prepared
The first gained of encapsulation rate is only 21.52%, and the lipid rate after separation is also only 77.78%, and later stage test gained Zeta electricity
Position is low, and stability is poor.
As preferred embodiment, the Raltitrexed pH sensitive liposomes are counted, including in parts by weight with the following group
Point：5 parts of Raltitrexed, 15 parts of phosphatidyl-ethanolamine, cholesterol succinate 35 or 40 parts, under this component, the present invention is made
Liposome more excellent effect is all had on encapsulation rate, stability, grain size.
The invention also discloses a kind of methods preparing Raltitrexed pH sensitive liposomes, it is characterised in that the method packet
Include following steps：
1) phosphatidyl-ethanolamine and cholesterol succinate are weighed in proportion, is dissolved in organic solvent, are uniformly mixed；
2) mixed liquor obtained by step 1) is placed in Rotary Evaporators, vacuum rotary steam removal is organic molten at 40 DEG C~55 DEG C
Agent makes it form one layer of uniform lipid membrane in bottle wall, and the sodium dihydrogen phosphate for the Raltitrexed that pH is 8~9 is then added
Buffer solution, at a certain temperature aquation certain time, filtering, freeze thawing obtain Raltitrexed pH sensitive liposome suspensions 3 times；
3) suspension obtained by step 2) is added in bag filter, the mistake in the phosphate sodium dihydrogen buffer solution dialysis of certain pH
Night obtains Raltitrexed pH sensitive liposomes.
The selection of preparation method also plays conclusive influence to liposome quality, sensitive in Lu Shan China fir docetaxels pH
Research [D] the Shandong University of liposome finds in 2014. researchs, using docetaxel pH sensitivity fat made from film hydration method
Plastid encapsulation rate highest is made encapsulation rate than freeze-thaw method and is higher by 30%；Due to Raltitrexed and a kind of anticarcinogen, so invention
People is during initially preparing Raltitrexed pH sensitive liposomes, it has therefore been attempted to it is prepared using film hydration method, however
The technique effect of gained is as shown in comparative example 3, and encapsulation rate is difficult to reach the expection of inventor, and inventor has attempted reverse steaming again
Prepared by hair method, as shown in comparative example 2, the encapsulation rate only up to 11.81% measured for the first time finally enables inventor not expect
, the encapsulation rates of the Raltitrexed pH sensitive liposomes obtained by freeze-thaw method only provided through the invention could keep just
Secondary gained encapsulation rate between 45.75%~49.95%, final gained encapsulation rate up to 89.45%~95.23% effect,
But according to comparative example 4 and comparative example 5 it is found that if pH conditions and vacuum rotary steam temperature to freeze-thaw method provided by the present invention into
Row, which changes, is also difficult to obtain encapsulation rate obtained by the present invention, or even will appear when pH is 7.5 and can not prepare liposome
Situation.
As preferred embodiment, organic solvent is chloroform in the step 1).
As preferred embodiment, hydration temperature is 40 DEG C in the step 2), and hydration time is 1 hour.
As preferred embodiment, filter method is using 0.22 μm of micro porous filtration in the step 2).
As preferred embodiment, the molecular cut off of bag filter is 1000Da in the step 3).
As preferred embodiment, the pH of phosphate sodium dihydrogen buffer solution is 7.4~9.0 in the step 3).
Advantageous effect
1. the Raltitrexed pH sensitive liposomes prepared by the present invention have excellent encapsulation rate, in material provided by the invention
For the first gained encapsulation rate of obtained liposome between 45.75%~49.95%, final gained encapsulation rate can under material and ratio
Up to 89.45%~95.23%.
2. the Raltitrexed pH sensitive liposomes prepared by the present invention have excellent quality, for the general blue-opalescent of white
Suspension liquid, and uniform particle diameter favorable dispersibility；Potential is between -46.6mv~-42.5mv, excellent in stability；Grain size exists
217nm~278nm, grain size is smaller and distribution is concentrated, and is suitable as the carrier of administration.
3. Raltitrexed pH sensitive liposomes produced by the present invention have excellent sensibility, in vitro in release experiment,
PH sensitive liposomes produced by the present invention in the dissolution medium of acidic environment, that is, pH5.0~6.5 of tumor tissues, release compared with
Soon, in the dissolution medium of normal structure pendular ring border, that is, pH7.4, release is slow, and pH sensitizations are apparent.
4. Raltitrexed pH sensitive liposomes produced by the present invention have excellent cancer resistance, inhibitor against colon carcinoma cells cell effect bright
It is aobvious, tumor tissues can be effectively targeted to；And blank liposome is non-toxic, reduces its side effect.
Description of the drawings
The grain size distribution of Raltitrexed pH sensitive liposomes prepared by Fig. 1 embodiments 2；
Raltitrexed pH sensitive liposomes prepared by Fig. 2 embodiments 2 are respectively 5.0,6.0,6.5 and 7.4 conditions in pH value
Under Accumulation dissolution curve graph；
The cell survival rate curve graph of Raltitrexed pH sensitive liposomes prepared by Fig. 3 embodiments 2 under various concentration.
Specific implementation mode
With reference to embodiments, the present invention will be described in further detail, it should be understood that specific reality described herein
It applies example to be only used to explain the present invention, be not intended to limit the present invention.
Embodiment 1
Justify in 100ml 1. precision weighs phosphatidyl-ethanolamine (DOPE) 35mg and cholesterol succinate (CHEMS) 15mg
In the flask of bottom, the dissolving of 5ml chloroforms is added, 40 DEG C of vacuum rotary steams remove organic solvent, it is made to form one layer in bottle wall uniformly
Lipid membrane；Phosphate sodium dihydrogen buffer solution (pH 8) 5ml of the Raltitrexed containing 5mg is added, 40 DEG C of aquations 1 hour use 0.22 μ
M filtering with microporous membrane, multigelation 3 times measure encapsulation rate to get thunder body song plug pH sensitive liposome suspensions；
2. taking 5ml suspensions to be added in the bag filter that molecular cut off is 1000Da, in the phosphoric acid of 1000ml pH 8.0
Dialysed overnight in sodium dihydrogen buffer solution, measures encapsulation rate again.
Embodiment 2
Justify in 100ml 1. precision weighs phosphatidyl-ethanolamine (DOPE) 40mg and cholesterol succinate (CHEMS) 15mg
In the flask of bottom, the dissolving of 5ml chloroforms is added, 45 DEG C of vacuum rotary steams remove organic solvent, it is made to form one layer in bottle wall uniformly
Lipid membrane；Phosphate sodium dihydrogen buffer solution (pH 9) 5ml of the Raltitrexed containing 5mg is added, 40 DEG C of aquations 1 hour use 0.22 μ
M filtering with microporous membrane, multigelation 3 times measure encapsulation rate to get thunder body song plug pH sensitive liposome suspensions；
2. taking 5ml suspensions to be added in the bag filter that molecular cut off is 1000Da, in the phosphoric acid of 1000ml pH8.0
Dialysed overnight in sodium dihydrogen buffer solution, measures encapsulation rate again.
Embodiment 3
Justify in 100ml 1. precision weighs phosphatidyl-ethanolamine (DOPE) 105mg and cholesterol succinate (CHEMS) 45mg
In the flask of bottom, the dissolving of 15ml chloroforms is added, 55 DEG C of vacuum rotary steams remove organic solvent, it is made to form one layer in bottle wall uniformly
Lipid membrane；Phosphate sodium dihydrogen buffer solution (pH 8.5) 15ml of the Raltitrexed containing 15mg is added, 40 DEG C of aquations 1 hour use
0.22 μm of filtering with microporous membrane, multigelation 3 times measure encapsulation rate to get thunder body song plug pH sensitive liposome suspensions；
2. taking 5ml suspensions to be added in the bag filter that molecular cut off is 1000Da, in the phosphoric acid of 1000ml pH 8.0
Dialysed overnight in sodium dihydrogen buffer solution, measures encapsulation rate again.Comparative example 1
Using preparation method same as Example 2 but phosphatidyl-ethanolamine is revised as soybean lecithin, by cholesterol amber
Amber acid esters is revised as cholesterol, and specific preparation process is as follows：
1. precision weighs soybean lecithin 40mg and cholesterol 15mg in 100ml round-bottomed flasks, addition 5ml chloroforms dissolve,
45 DEG C of vacuum rotary steams remove organic solvent, it is made to form one layer of uniform lipid membrane in bottle wall；Raltitrexed containing 5mg is added
Phosphate sodium dihydrogen buffer solution (pH8.5) 5ml, 40 DEG C of aquations 1 hour use 0.22 μm of filtering with microporous membrane, multigelation 3
It is secondary to get thunder body song plug pH sensitive liposome suspensions, measure encapsulation rate；
2. taking 5ml suspensions to be added in the bag filter that molecular cut off is 1000Da, in the phosphoric acid of 1000ml pH 8.0
Dialysed overnight in sodium dihydrogen buffer solution, measures encapsulation rate again.Comparative example 2
It is prepared using the material of weight ratio same as Example 1, but by reverse evaporation, preparation process is such as
Under：
1. precision weighs phosphatidyl-ethanolamine 35mg, cholesterol succinate 15mg in 100mL round-bottomed flasks, it is added
5ml chloroforms dissolve, and the phosphate sodium dihydrogen buffer solution 5ml of the 5mg containing Raltitrexed is added, and adjust pH to 8.5, and ultrasonic 8min makes shape
At uniform w/o type emulsion；40 DEG C of vacuum rotary steams remove chloroform, form lipid membrane on the wall；Continue to rotate, makes to be formed
Even Liposomal suspensions, 0.22 μm of filtering with microporous membrane obtain the liposome turbid liquor of uniform particle sizes, measure encapsulation rate；
2. taking 5ml suspensions to be added in the bag filter that molecular cut off is 1000, in the di(2-ethylhexyl)phosphate of 1000ml pH 8.0
Dialysed overnight in hydrogen sodium buffer solution, measures encapsulation rate again.Comparative example 3
It is prepared using the material of weight ratio same as Example 3, but by film hydration method, preparation process is such as
Under：
1. precision weighs phosphatidyl-ethanolamine 105mg and cholesterol succinate 45mg in 100mL round-bottomed flasks, it is added
15ml chloroforms dissolve, and 55 DEG C of vacuum rotary steams remove chloroform, it is made to form one layer of uniform lipid membrane in bottle wall；Addition is replaced containing thunder
The sodium dihydrogen phosphate 15ml of song plug 15mg, adjusts pH to 8.5,40 DEG C of aquations 1 hour, 0.22 μm of filtering with microporous membrane, i.e.,
Thunder body song plug pH sensitive liposome suspensions are obtained, encapsulation rate is measured；
2. taking 5ml suspensions to be added in the bag filter that molecular cut off is 1000Da, in the phosphoric acid of 1000ml pH 8.0
Dialysed overnight in sodium dihydrogen buffer solution, measures encapsulation rate again.Comparative example 4
It is 60 DEG C using the material and preparation method of weight ratio same as Example 3, but by vacuum rotary steam temperature modification,
Preparation process is as follows：
1. precision weighs phosphatidyl-ethanolamine 105mg and cholesterol succinate 45mg in 100mL round-bottomed flasks, it is added
15ml chloroforms dissolve, and 60 DEG C of vacuum rotary steams remove chloroform, it is made to form one layer of uniform lipid membrane in bottle wall；Addition is replaced containing thunder
The sodium dihydrogen phosphate 15ml of song plug 15mg, adjusts pH to 8.5,40 DEG C of aquations 1 hour, 0.22 μm of filtering with microporous membrane, i.e.,
Thunder body song plug pH sensitive liposome suspensions are obtained, encapsulation rate is measured；
2. taking 5ml suspensions to be added in the bag filter that molecular cut off is 1000Da, in the phosphoric acid of 1000ml pH 8.0
Dialysed overnight in sodium dihydrogen buffer solution, measures encapsulation rate again.Comparative example 5
Using the material and preparation method of weight ratio same as Example 3, but by the di(2-ethylhexyl)phosphate of the 15mg containing Raltitrexed
The pH of hydrogen sodium solution is adjusted to 7.5, and preparation process is as follows：
1. precision weighs phosphatidyl-ethanolamine 105mg and cholesterol succinate 45mg in 100mL round-bottomed flasks, it is added
15ml chloroforms dissolve, and 20 DEG C of vacuum rotary steams remove chloroform, it is made to form one layer of uniform lipid membrane in bottle wall；Addition is replaced containing thunder
The sodium dihydrogen phosphate 15ml of song plug 15mg, adjusts pH to 7.5,40 DEG C of aquations 1 hour, 0.22 μm of filtering with microporous membrane, i.e.,
Thunder body song plug pH sensitive liposome suspensions are obtained, encapsulation rate is measured；
2. taking 5ml suspensions to be added in the bag filter that molecular cut off is 1000Da, in the phosphoric acid of 1000ml pH 8.0
Dialysed overnight in sodium dihydrogen buffer solution, measures encapsulation rate again.Experimental example 1
Above-described embodiment and comparative example are measured into gained encapsulation rate and are shown in Table 1, the assay method of the encapsulation rate is：Glucan
Exclusion chromatography；
The entrapment efficiency determination result of the different embodiments of table 1
| Project | Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 | Comparative example 5 |
| Measure encapsulation rate for the first time | 46.05% | 49.95% | 45.75% | 21.52% | 11.81% | 25.25% | 19.35% | It can not prepare |
| Encapsulation rate is measured again | 93.27% | 95.23% | 89.45% | 77.78% | 81.21% | 78.34% | 73.43% | It can not prepare |
Encapsulation rate refers to the percentage amounts for being wrapped substance (such as certain drug) and accounting for drug total amount in Liposomal suspensions, liposome
It is to judge the whether effective major criterion of method for preparing lipidosome that encapsulation rate before separation measures encapsulation rate for the first time, according to table 1
It is found that if pH sensitivity membrane materials are replaced with soybean lecithin and cholesterol, the encapsulation rate of first gained is reduced to embodiment 1
The 43.08%~47.03% of~3；And if the preparation method of liposome is replaced, the encapsulation rate of liposome is also difficult to
To guarantee, the liposome prepared using reverse evaporation, the encapsulation rate measured for the first time is reduced to Examples 1 to 3
23.64%~25.81%, using the liposome of film hydration method preparation, its encapsulation rate measured for the first time is reduced to embodiment 1
The 50.55%~55.19% of~3 is all difficult to it follows that pH sensitivity membrane materials to be replaced or be replaced preparation method
Obtain the getable encapsulation rate of present invention institute.
According to comparative example 4 it is found that if being 60 DEG C by the vacuum rotary steam temperature modification of liposome is prepared, measure for the first time
Encapsulation rate be reduced to the 38.73%~42.30% of Examples 1 to 3, if the pH of Raltitrexed disodium hydrogen phosphate changed
It is 7.5, then the liposomal particle size prepared is huge or liposome membrane can not elute, and can not form liposome, it follows that such as
Fruit modifies to technique provided by the present invention, then is equally difficult to ensure the encapsulation rate of liposome.
Experimental example 2
Quality evaluation is carried out to Raltitrexed pH sensitive liposomes made from all experimental examples 1~3, by 2 gained of embodiment
Grain size be prepared as grain size distribution, the distribution map is shown in Fig. 1；Embodiment 1 is similar to 3 acquired results of embodiment.
2 quality evaluation of table
According to table 2 it is found that Raltitrexed pH sensitive liposomes prepared by the present invention are the suspension of the general blue-opalescent of white
Body, and uniform particle diameter favorable dispersibility, and as Zeta potential absolute value ＞ 30mv, show that liposome has good stabilization
Property, and Zeta potential absolute value more high stability is better, lipid bulk potential produced by the present invention -46.6mv~-42.5mv it
Between, excellent in stability；Liposomal particle size is made in 217nm~278nm in the present invention, and grain size is smaller and distribution is concentrated, and is suitable as
The carrier of administration.
Experimental example 3
Vitro release experiment is carried out to experimental example 1~3；It is as follows：
Prepare pH be respectively 5.0,6.0,6.5,7.4 phosphate buffer 25mL as dissolution medium, then take centainly
Drug-loaded liposome is measured in the bag filter that molecular cut off is 1000Da, places it in the dissolution medium of above-mentioned difference pH, adopts
With constant temperature oscillation method, respectively at 0.5h, 1h, 2h, 4h, 6h, 8h, 12h, for 24 hours, 48h sample 1mL, fluid infusion 1mL, calculate accumulation and release
Degree of putting, Examples 1 to 3 drug-loaded liposome release rule in different pH medium is different, is in the acidic environment of tumor tissues
In the dissolution medium of pH5.0~6.5, release is very fast, and in the dissolution medium of normal structure pendular ring border, that is, pH7.4, release is slow,
PH sensitizations are apparent.
It is similar to 3 acquired results of embodiment that 2 release profiles of embodiment are drawn into such as Fig. 2, embodiment 1.
Embodiment 4
Cytotoxicity experiment is carried out to experimental example 1~3, is as follows：
The HCT-116 cells fetched to growth period carry out postdigestive cell suspension according to 2000, every hole cell dilute
It releases.It is cultivated 24 hours as in cell incubator per hole 100uL in 96 orifice plate inoculating cell suspensions.Configuring drug concentration is
The drug-loaded liposome solution of 16ug/mL, using coubling dilution so that the drug concentration of drug-loaded liposome group is respectively 16ug/
ML, 8ug/mL, 4ug/mL, 2ug/mL, 1ug/mL, 0.5ug/mL, 0.25ug/mL, 0.125ug/mL, 0.0625ug/mL, blank
Liposome group is the solution of corresponding concentration of liposomes.100uL, parallel 5 hole of each concentration, in cell incubator are added per hole
Interior culture 72h.10uL CCK-8 solution is added after culture per hole, 2h is cultivated in cell incubator, is measured with microplate reader
Absorption value at 450nm calculates its cell survival rate.The result shows that within the scope of a certain concentration, blank liposome is non-toxic,
And drug-loaded liposome inhibitor against colon carcinoma cells cell is with obvious effects, it is known that it can effectively reduce its toxic side effect.
It is similar to 3 acquired results of embodiment that 2 cell survival rate trend of embodiment is drawn into such as Fig. 3, embodiment 1.
Claims (8)
1. a kind of Raltitrexed pH sensitive liposomes, which is characterized in that the Raltitrexed pH sensitive liposomes, in parts by weight
Meter, including following components：5~15 parts of Raltitrexed, 35~105 parts of phosphatidyl-ethanolamine, 15~45 parts of cholesterol succinate.
2. a kind of Raltitrexed pH sensitive liposomes according to claim 1, which is characterized in that the Raltitrexed pH is quick
Feel liposome, counts in parts by weight, including following components：5 parts of Raltitrexed, 15 parts of phosphatidyl-ethanolamine, cholesterol succinic acid
Ester 35 or 40 parts.
3. a kind of preparing the method such as claim 1-2 any one of them Raltitrexed pH sensitive liposomes, which is characterized in that
It the described method comprises the following steps：
1) phosphatidyl-ethanolamine and cholesterol succinate are weighed in proportion, is dissolved in organic solvent, are uniformly mixed；
2) mixed liquor obtained by step 1) is placed in Rotary Evaporators, vacuum rotary steam removes organic solvent at 40 DEG C~55 DEG C, makes
It forms one layer of uniform lipid membrane in bottle wall, and the sodium dihydrogen phosphate buffering for the Raltitrexed that pH is 8~9 is then added
Liquid, at a certain temperature aquation certain time, filtering, freeze thawing obtain Raltitrexed pH sensitive liposome suspensions 3 times；
3) suspension obtained by step 2) is added in bag filter, in the phosphate sodium dihydrogen buffer solution dialysis of certain pH overnight,
Obtain Raltitrexed pH sensitive liposomes.
4. preparation method according to claim 3, which is characterized in that organic solvent is chloroform in the step 1).
5. preparation method according to claim 3, which is characterized in that hydration temperature is 40 DEG C in the step 2), aquation
Time is 1 hour.
6. preparation method according to claim 3, which is characterized in that filter method is using 0.22 μm in the step 2)
Micro porous filtration.
7. preparation method according to claim 3, which is characterized in that the molecular cut off of bag filter is in the step 3)
1000Da。
8. preparation method according to claim 3, which is characterized in that the pH of phosphate sodium dihydrogen buffer solution in the step 3)
It is 7.4~9.0.
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| CN201810682308.7A CN108619098B (en) | 2018-06-27 | 2018-06-27 | Raltitrexed pH sensitive liposome and preparation method thereof |
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| CN116350589A (en) * | 2023-03-27 | 2023-06-30 | 安徽农业大学 | Violacein pH sensitive liposome and preparation method and application thereof |
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| CN105287436A (en) * | 2014-05-29 | 2016-02-03 | 邢蓉 | Raltitrexed lipid nano-carrier and method for preparing same |
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| SERGIO SIMOES ETAL: "On the formulation of pH-sensitive liposomes with long circulation times", 《ADVANCED DRUG DELIVERY REVIEWS》 * |
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| CN116350589A (en) * | 2023-03-27 | 2023-06-30 | 安徽农业大学 | Violacein pH sensitive liposome and preparation method and application thereof |
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