CN109730975A - A method of it maintains to carry medicine red blood cell bioactivity while improving its drugloading rate - Google Patents
A method of it maintains to carry medicine red blood cell bioactivity while improving its drugloading rate Download PDFInfo
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Abstract
The invention belongs to field of pharmaceutical preparations, and in particular to a method of it maintains to carry medicine red blood cell bioactivity while improving drugloading rate.The method at least includes the following steps: red blood cell being added into hypotonic glucan medical fluid, is incubated for for the first time；High sepage is added, second of incubation makes the purpose drug encapsulation in hypotonic glucan medical fluid in red blood cell, and centrifuge washing removes glucan, obtains carrying medicine red blood cell.Easy using method provided by the invention, efficiently, effect is good, and the last different kind organism activity index obtained for carrying medicine red blood cell and natural red blood cell do not have significant difference, has been properly arrived at maintenance and has carried the active purpose of medicine red blood cell external biological；While maintaining bioactivity, method provided by the invention can also improve the amount of containing of drug.
Description
Technical field
The invention belongs to field of pharmaceutical preparations, and in particular to a kind of maintenance carries medicine red blood cell bioactivity and improves its load simultaneously
The method of dose.
Background technique
Red blood cell (RBC) is the most abundant cell type of human body, accounts for about total cell number a quarter, in mature erythrocyte not
Containing substances such as organelle, nucleic acid, content ingredient is simple.Circulation time is long in vivo for red blood cell, and wherein people RBC maturation is laggard
Entering the circulatory system can survive 100-120 days, and Rat Erythrocytes can survive 50-60 days.These characteristics based on red blood cell, i.e. source
Abundant, good biocompatibility, good biodegradability do not induce immune response, circulation time in vivo long etc., from upper
Age in century 60-70, red blood cell are just widely used in containing for each substance, including enzyme, nucleic acid and small point various
Sub- drug.However when the red blood cell of drug incorporation outside perfect aspect reenters body-internal-circulation, the service life often only has neutral red thin
The 1/3 of born of the same parents is even shorter, at present still without result of the above problems.
Summary of the invention
The object of the present invention is to provide a kind of methods for maintaining to carry medicine red blood cell bioactivity while improving its drugloading rate, can
Farthest to protect red blood cell to reduce bioactivity from the influence of external various unfavorable factors, while can also improve
Drugloading rate.To achieve the goals above and other related purposes, the present invention adopts the following technical scheme:
The first aspect of the present invention provides a kind of side for maintaining to carry medicine red blood cell bioactivity while improving its drugloading rate
Method at least includes the following steps:
(1) red blood cell is added into hypotonic glucan medical fluid, is incubated for for the first time；
(2) high sepage is added in the mixture after being incubated for into step (1), second of incubation makes hypotonic glucan medical fluid
In purpose drug encapsulation in red blood cell, centrifuge washing remove glucan, obtain carry medicine red blood cell.
In step (1), the hypotonic glucan medical fluid refers to, containing glucan, sodium chloride and needs to be carried on red blood cell
On purpose drug hypotonic aqueous solution.
The glucan has water solubility.Optionally, the water solubility of the glucan is good.
The molecular weight of the glucan is 10000-70000.
In one embodiment, in the hypotonic glucan medical fluid, on the basis of the hypotonic glucan medical fluid total amount
Meter, the mass fraction of the glucan are 2%-20%.
In one embodiment, it is counted on the basis of the hypotonic glucan medical fluid total amount, the hypotonic glucan medical fluid
Constituent include:
Purpose drug 1mg/mL-10mg/mL,
Mass fraction is the sodium chloride of 0.4%-0.6%,
Mass fraction is the glucan of 2%-20%,
Solvent is water.
In one embodiment, the purpose drug be selected from betamethasone sodium phosphate, dexamethasone sodium phosphate, hydrochloric acid Ah
Mycin, sodium artesunate, morphine hydrochloride and bovine serum albumin(BSA).
In one embodiment, in step (1), the red blood cell and hypotonic Portugal are poly-
The volume ratio of sugared medical fluid is 1:(4-20).
In one embodiment, in step (1), the red blood cell is packed red cells.
Further, the preparation method of the packed red cells at least includes the following steps: animal's whole blood centrifuge separation is gone
Except blood plasma and leucocyte, the packed red cells are obtained after washing.
Animal's whole blood centrifugal condition are as follows: 4 DEG C, 3800r/min.
The animal is mammal, can be rodent, primate.Such as can be monkey, rabbit, goat,
Rat.
In one embodiment, in step (1), the temperature of first time incubation are as follows: 0-10 DEG C.It optionally, is 0-4 DEG C.
In one embodiment, in step (1), the time being incubated for for the first time is 10-60min.
Further, it in step (2), is counted on the basis of the high sepage total amount, the constituent of the high sepage includes:
Mass fraction is 2.5-5.5% sodium chloride, 1-10mg/ml Sodium Pyruvate and 2-20mg/ml glucose, and solvent is water.
In one embodiment, in step (2), the volume ratio of high sepage and hypotonic glucan medical fluid is 1:(4-20).
In one embodiment, in step (2), second of temperature being incubated for are as follows: 30-40 DEG C.
In one embodiment, in step (2), second of the time being incubated for is 10-60min.
Second aspect of the present invention provides preceding method and is used to prepare the purposes for carrying medicine red cell preparation.
Compared with prior art, the present invention have it is following the utility model has the advantages that
1) easy using method provided by the invention, efficiently, effect is good, the last different kind organism obtained for carrying medicine red blood cell
Activity index and natural red blood cell do not have significant difference, have been properly arrived at maintenance and have carried the active mesh of medicine red blood cell external biological
's.
2) while maintaining bioactivity, method provided by the invention can also improve the amount of containing of drug.
Detailed description of the invention
Upon reading the detailed description of non-limiting embodiments with reference to the following drawings, other feature of the invention,
Objects and advantages will be more obvious:
Fig. 1 is the influence lab diagram for investigating different molecular weight, various concentration glucan to red blood cell drugloading rate；
Fig. 2 is to investigate different molecular weight, various concentration glucan to the influence lab diagram for carrying medicine osmotic fragility；
Fig. 3 is the influence for investigating different molecular weight, various concentration glucan to medicine red blood cell Na+/K+-ATP enzymatic activity is carried
Lab diagram；
Fig. 4 A is the influence lab diagram investigating glucan and turning up to phosphatidylserine on load medicine erythrocyte membrane；For neutral red
The FSC-SSC streaming figure of cell negative control group (Negative Control)；
Fig. 4 B is the influence lab diagram investigating glucan and turning up to phosphatidylserine on load medicine erythrocyte membrane；For neutral red
The FSC-SSC streaming figure of cell (NRBC)；
Fig. 4 C is the influence lab diagram investigating glucan and turning up to phosphatidylserine on load medicine erythrocyte membrane；For Portugal is not added
The glycan FSC-SSC streaming figure obtained for carrying medicine red blood cell (BSP-RBC)；
Fig. 4 D is the influence lab diagram investigating glucan and turning up to phosphatidylserine on load medicine erythrocyte membrane；For Portugal is added
Glycan (T40-10%, it is 40000 that wherein T40, which represents the molecular weight of glucan, and 10% represents the mass fraction of glucan) is made afterwards
The FSC-SSC figure of the load medicine red blood cell obtained；
Fig. 4 E is the influence lab diagram investigating glucan and turning up to phosphatidylserine on load medicine erythrocyte membrane；For phosphatidyl
Serine turns up experimental result picture.
Fig. 5 is to investigate glucan to the influence lab diagram for carrying medicine red cell morphology structure；Wherein: 5A NRBC, 5B are
BSP-RBC, 5C T40-10%-BSP-RBC.
* is represented in each figure, is compared with BSP-RBC group, and p < 0.05 shows significant difference；# is represented, with NRBC group pair
Than p < 0.05 shows significant difference.(the pairs of t of Spss software is examined).
Specific embodiment
Before further describing the specific embodiments of the present invention, it should be appreciated that protection scope of the present invention is not limited to down
State specific specific embodiment；It is also understood that term used in the embodiment of the present invention is specific specific in order to describe
Embodiment, rather than limiting the scope of protection of the present invention.The test method of actual conditions is not specified in the following example,
Usually according to normal condition, or according to condition proposed by each manufacturer.
When embodiment provides numberical range, it should be appreciated that except non-present invention is otherwise noted, two ends of each numberical range
Any one numerical value can be selected between point and two endpoints.Unless otherwise defined, the present invention used in all technologies and
Scientific term is identical as the normally understood meaning of those skilled in the art of the present technique.Except specific method, equipment used in embodiment,
Outside material, grasp and record of the invention according to those skilled in the art to the prior art can also be used and this
Any method, equipment and the material of the similar or equivalent prior art of method described in inventive embodiments, equipment, material come real
The existing present invention.
Unless otherwise stated, disclosed in this invention experimental method, detection method, preparation method be all made of this technology neck
Molecular biology, biochemistry, chromatin Structure and the analysis of domain routine, analytical chemistry, cell culture, recombinant DNA technology and
The routine techniques of related fields.
The addition of 1 glucan of embodiment and the corresponding preparation for carrying medicine red blood cell
(glucan is added) in experimental group:
(1) taking weight is the male SD rat of 200g or so, carries out posterior orbit veniplex with the capillary of diameter 0.5mm and takes
Blood, each amount for taking blood are 0.2-0.3ml.4 DEG C, 2000r/min is centrifuged 4min, abandons supernatant, removes blood plasma and leukocytic cream, then
Brine 1-2 times of 1ml pre-cooling is added to supernatant close to colourless, obtains lower layer's packed red cells.
(2) it takes 100 μ l Washed Red Blood Cells to be added in the hypotonic glucan medical fluid of 400 μ l, is gently mixed uniformly, 4 DEG C of standings
It is incubated for 20min, during which can gently vibrate mixing every 4min.The wherein composition of hypotonic glucan medical fluid are as follows: mass fraction is
0.45% sodium chloride, 4mg/ml betamethasone sodium phosphate, glucan, solvent are water.(it is different that different molecular weight is provided in experiment
The glucan of concentration, including molecular weight be 10000, mass fraction be respectively 2%, 5%, 10% T10-2%, T10-5%,
T10-10%；Molecular weight is 40000, and mass fraction is respectively 2%, 5%, 10% T40-2%, T40-5%, T40-10%；
Molecular weight is 70000, and mass fraction is respectively 0.5%, 1%, 2% T70-0.5%, T70-1%, T70-2%.)
(3) 100 μ l high sepages are added in system in (2), are gently mixed uniformly, 37 DEG C of stationary incubation 30min during which can be every
Mixing is gently vibrated every 5min.The composition of its middle and high infiltration liquid are as follows: mass fraction is 4.5% sodium chloride, 100mM glucose, 100mM
Sodium Pyruvate, solvent are water.
(4) 4 DEG C, 2000r/min is centrifuged 4min, discards supernatant, then with brine 3 times of 1ml pre-cooling, remove and add
The glucan and free drug entered, obtains the load medicine red blood cell protected through glucan.
Blank control group:
The control group carries medicine red blood cell BSP-RBC and prepares by above-mentioned (1)-(4) the method, only difference is that,
Glucan is free of in the hypotonic medical fluid.
Embodiment 2. investigates the influence of different molecular weight, various concentration glucan to medicine red blood cell drugloading rate is carried
The supernatant of each step in embodiment 1 (4) to be collected, 200 μ l are drawn, the methanol that 5 times of i.e. 1ml of volume are added mixes,
Precipitate a small amount of glucan and hemoglobin, after 0.22 μm of disposable membrane filtration, high performance liquid chromatograph sample introduction measures supernatant
The content of betamethasone sodium phosphate in liquid.Betamethasone phosphorus in drugloading rate=addition betamethasone sodium phosphate total amount-supernatant
Sour sodium content.
The wherein determination condition of high performance liquid chromatograph are as follows:
Chromatographic column: AgilentXB-C18(5μm,4.6×250nm)；Detector: Agilent
Technologics UV detector；Mobile phase: methanol -0.05mol/l potassium dihydrogen phosphate (1:1)；Flow velocity: 1ml/min；Column
Temperature: 40 DEG C；Detection wavelength: 254nm.
Fig. 1 is the results show that the drugloading rate of red blood cell can be improved in the addition of glucan.Compared to being not added made from glucan
Medicine red blood cell BSP-RBC is carried, wherein the load medicine obtained after joined T10-5%, T10-10%, T40-5%, T40-10% is red thin
Born of the same parents, drugloading rate, which has, to be obviously improved.* p < 0.05 is represented, there is significant difference.
Embodiment 3. investigates the influence of different molecular weight, various concentration glucan to medicine osmotic fragility is carried
By such as the following table 1, a series of sodium chloride solution of different osmotics is prepared with 1%NaCl and ultrapure water.Take 10 parts of realities
The load medicine red blood cell obtained of experimental group in example 1 is applied, the sodium chloride solution of 4 times of volume different osmotics, 4 DEG C of standings are separately added into
2h, 4 DEG C, 2000r/min is centrifuged 5min, and 100 μ l of supernatant is taken to be added in 96 orifice plates, and 3 parallel holes are arranged in every part of sample, uses enzyme
Mark absorbance of the instrument measurement various kinds sample wells at 540nm.Hemolysis rate is calculated according to following formula, it is molten using osmotic pressure as abscissa
Blood rate is ordinate, draws osmotic fragility curve.Parallel test 3 times, natural red blood cell, blank control group BSP-RBC load medicine are red
The osmotic fragility measuring method of cell is identical with this.
Table 1
Fig. 2 the results show that the addition of glucan can be enhanced carry medicine red blood cell confrontation osmotic pressure ability, that is, enhance load
The toughness of medicine red blood cell.Wherein the improvement of T40-10% is especially pronounced, and this glucan load medicine red blood cell obtained is added
Osmotic fragility and natural red blood cell it is almost consistent.
Embodiment 4. investigates different molecular weight, various concentration glucan to load medicine red blood cell Na+/K+The shadow of atpase activity
It rings
It takes the experimental group medicine red blood cell obtained that carries in 20 μ l embodiments 1 to add the 180 ultrapure water-splittings of μ l, stands 30min, obtain
Transparent lysate.It takes the 20 transparent lysates of μ l that ultrapure water is added to dilute 40 times to lysate in colourless or micro mist color transparence, uses BCA
Protein concentration quantification kit measures protein concentration, then measures transparent lysate according to the step of ultramicron ATP enzyme kit
Middle Na+/K+The activity of ATP enzyme.The activity of Na+/K+-ATP enzyme on BSP-RBC film is finally converted by corresponding proportion.Neutral red
Cell, blank control group BSP-RBC carry medicine red blood cell Na+/K+The measuring method of atpase activity is identical with this.
Fig. 3 carries medicine red blood cell Na the results show that the addition of glucan can protect+/K+The activity of ATP enzyme.Wherein T40-
5%, the protecting effect of T40-10%, T70-1% and T70-2% are especially pronounced.
Embodiment 5. investigates the influence that glucan turns up to phosphatidylserine on load medicine erythrocyte membrane
Take in 10 μ l embodiments 1 experimental group T40-10%-RBC group to carry medicine red blood cell into streaming pipe, be added 490 μ l 1 ×
PBS dilution, then the load medicine red blood cell suspension after 10 μ l dilution is taken, 100 μ l 1 × binding buffer are added, mix well
Afterwards, 5 μ l Annexin-V-FITC are added, room temperature, which is protected from light, is incubated for 10-15min.400 μ l 1 × binding buffer are added, fill
Divide and mixes, loading measurement.One negative control group of experimental setup (Negative Control), a neutral red groups of cells
(NRBC), a blank control group carries medicine red blood cell (BSP-RBC) and T40-10%-RBC group.Wherein negative control group is not
Add 5 μ l Annexin-V-FITC, directly adds 500 μ 1 × binding of l buffer；Neutral red groups of cells, blank control
Group BSP-RBC carries the processing step of medicine red blood cell with T40-10%-RBC group.
Fig. 4 A, 4B, 4C, 4D are the results show that Negative Control, NRBC, BSP-RBC, T40-10%-RBC exist
Distribution in FSC-SSC is almost the same, illustrates that the addition of glucan will not influence and carries the form of medicine red blood cell and thin with neutral red
The form of born of the same parents is almost the same.For Fig. 4 E the results show that the FITC of BSP-RBC group is most strong, FITC positive ratio is 26%, i.e. phosphatidyl
Serine rate of turning up is 26%, and T40-10%-RBC group FITC is obviously moved to left, and FITC positive ratio is reduced to 7.9%, i.e. phosphatide
Acyl serine turn up rate be 7.9%.The two compares, and p value shows significant difference less than 0.05.Illustrate the Portugal T40-10%
The addition of glycan can reduce turning up for phosphatidylserine on erythrocyte membrane, have good protective effect to red blood cell.
Embodiment 6. investigates influence of the glucan to medicine red cell morphology is carried
Experimental group T40-10%-RBC group in 50 μ L embodiments 1 is taken to carry the addition of medicine red blood cell equipped with 2.5% glutaraldehyde of 1mL
EP pipe in, shake gently mixing, fixed 30min, during which gently shaking EP pipe every 5min prevents red blood cell to be largely deposited in bottom
Portion.The red blood cell suspension fixed is centrifuged 4min with the revolving speed of 2000r/min, removes supernatant, and with 1 × PBS buffer solution
It cleans 1-2 times and removes extra fixer.Erythroprecipitin after cleaning is added to equipped with (0.4% Gao Meng of fixer after 1mL
Sour potassium, 0.6% potassium bichromate) EP pipe in, shake gently mixing, suspend fixed 5min.The red blood cell suspension that will be fixed
It is centrifuged 4min with the revolving speed of 2000r/min, removes supernatant, and clean 1-2 times with ultrapure water and remove extra rear fixer.Then
With the ethanol dehydration agent (30%, 50%, 70%, 80%, 85%, 90%, 95%, 100%) of configured various concentration to red
Cell carries out serial dehydration.The dehydration that suspends of the ethyl alcohol of each concentration is primary, each 5min, then with the revolving speed of 2000r/min from
Heart 4min, removes supernatant, carries out the ethanol dehydration of next concentration, wherein 100% ethanol dehydration is twice.It finally will be after centrifugation
Cell, retains 1/3 supernatant, and piping and druming uniformly, drips on filter paper dick, is put into vacuum oven, takes out sample after 10min.It dips in
Take a small amount of sample powder on conducting resinl, gold-plated, scanning electron microscope observation.Natural red blood cell, blank control group BSP-
RBC carries medicine red blood cell processing method and is identical with this.
Fig. 5 is the results show that the addition of glucan will not influence the form for carrying medicine red blood cell and the form with natural red blood cell
It is almost the same, it is all biconcave round pie.
Embodiment 7
The present invention also refers to 1 experimental group of embodiment, prepares carry medicine red blood cell otherwise, and crisp to drugloading rate, infiltration
Property, red blood cell Na+/K+Phosphatidylserine turns up on atpase activity, film, form is characterized.
Method 1, with 1 experimental group of embodiment the difference is that: in step (2), 100 μ l Washed Red Blood Cells is taken to be added to
It in the hypotonic glucan medical fluid of 2000 μ l, is gently mixed uniformly, 0 DEG C of stationary incubation 10min, during which can gently be vibrated every 2min mixed
It closes.The wherein composition of hypotonic glucan medical fluid are as follows: mass fraction is 0.4% sodium chloride, 4mg/ml dexamethasone sodium phosphate, molecule
Amount is 70000, and the glucan that mass fraction is 20%, solvent is water；In step (3), it is hypertonic that 100 μ l are added in system in (2)
Liquid is gently mixed uniformly, during which 30 DEG C of stationary incubation 60min can gently vibrate mixing every 5min.The composition of its middle and high infiltration liquid
Are as follows: mass fraction is 2.5% sodium chloride, 2mg/ml glucose, 10mg/ml Sodium Pyruvate, and solvent is water.Remaining is all identical.
Method 2, with 1 experimental group of embodiment the difference is that: in step (2), 100 μ l Washed Red Blood Cells is taken to be added to
In the hypotonic glucan medical fluid of 1000 μ l, it is gently mixed uniformly, during which 10 DEG C of stationary incubation 60min can gently be vibrated every 2min
Mixing.The wherein composition of hypotonic glucan medical fluid are as follows: mass fraction is 0.6% sodium chloride, 10mg/ml doxorubicin hydrochloride, molecule
Amount is 60000, and the glucan that mass fraction is 15%, solvent is water；In step (3), it is hypertonic that 100 μ l are added in system in (2)
Liquid is gently mixed uniformly, during which 40 DEG C of stationary incubation 10min can gently vibrate mixing every 1min.The composition of its middle and high infiltration liquid
Are as follows: mass fraction is 5.5% sodium chloride, 20mg/ml glucose, 1mg/ml Sodium Pyruvate, and solvent is water.Remaining is all identical.
Method 3, with 1 experimental group of embodiment the difference is that: in step (2), 100 μ l Washed Red Blood Cells is taken to be added to
It in the hypotonic glucan medical fluid of 1200 μ l, is gently mixed uniformly, 5 DEG C of stationary incubation 30min, during which can gently be vibrated every 2min mixed
It closes.The wherein composition of hypotonic glucan medical fluid are as follows: mass fraction is 0.5% sodium chloride, 1mg/ml sodium artesunate, molecular weight are
40000, the glucan that mass fraction is 2%, solvent is water；In step (3), 100 μ l high sepages are added in system in (2), gently
Light to be uniformly mixed, during which 35 DEG C of stationary incubation 40min can gently vibrate mixing every 5min.The composition of its middle and high infiltration liquid are as follows: matter
Amount score is 3% sodium chloride, 10mg/ml glucose, 5mg/ml Sodium Pyruvate, and solvent is water.Remaining is all identical.
As a result: load medicine red blood cell obtained has following characteristics in method 1, method 2 and method 3: the addition of glucan mentions
The high drugloading rate of red blood cell, is added the load medicine red blood cell prepared after T10-5%, T10-10%, T40-5%, T40-10%, carries
Dose, which has, is obviously improved (p < 0.05).The addition of glucan enhances the ability for carrying medicine red blood cell confrontation osmotic pressure,
Wherein the improvement of T40-10% is especially pronounced, and this glucan osmotic fragility obtained for carrying medicine red blood cell and equal day is added
Right red blood cell is almost consistent.The addition of glucan, which protects, carries medicine red blood cell Na+/K+The activity of ATP enzyme, wherein T40-5%,
The protecting effect of T40-10%, T70-1% and T70-2% are especially pronounced.The addition of T40-10% glucan reduces red thin
Phosphatidylserine turns up on after birth, to red blood cell has good protective effect, and will not influence the form for carrying medicine red blood cell,
It is almost the same with the form of natural red blood cell, it is all biconcave round pie.
The above, only presently preferred embodiments of the present invention, not to the present invention in any form with substantial limitation,
It should be pointed out that under the premise of not departing from the method for the present invention, can also be made for those skilled in the art
Several improvement and supplement, these are improved and supplement also should be regarded as protection scope of the present invention.All those skilled in the art,
Without departing from the spirit and scope of the present invention, when made using disclosed above technology contents it is a little more
Dynamic, modification and the equivalent variations developed, are equivalent embodiment of the invention；Meanwhile all substantial technologicals pair according to the present invention
The variation, modification and evolution of any equivalent variations made by above-described embodiment, still fall within the range of technical solution of the present invention
It is interior.
Claims (10)
1. a kind of maintain to carry the method that medicine red blood cell bioactivity improves its drugloading rate simultaneously, at least include the following steps:
(1) red blood cell is added into hypotonic glucan medical fluid, is incubated for for the first time；
(2) high sepage is added in the mixture after being incubated for into step (1), second of incubation makes in hypotonic glucan medical fluid
For purpose drug encapsulation in red blood cell, centrifuge washing removes glucan, obtains carrying medicine red blood cell.
2. as described in claim 1 maintain to carry the method that medicine red blood cell bioactivity improves its drugloading rate simultaneously, feature exists
In in step (1), the hypotonic glucan medical fluid refers to, containing glucan, sodium chloride and needs to be carried on the mesh on red blood cell
Drug hypotonic aqueous solution.
3. as claimed in claim 2 maintain to carry the method that medicine red blood cell bioactivity improves its drugloading rate simultaneously, feature exists
In the molecular weight of the glucan is 10000-70000.
4. as claimed in claim 2 maintain to carry the method that medicine red blood cell bioactivity improves its drugloading rate simultaneously, feature exists
In being counted on the basis of the hypotonic glucan medical fluid total amount, the mass fraction of the glucan in the hypotonic glucan medical fluid
For 2%-20%.
5. as claimed in claim 2 maintain to carry the method that medicine red blood cell bioactivity improves its drugloading rate simultaneously, feature exists
In being counted on the basis of the hypotonic glucan medical fluid total amount, the constituent of the hypotonic glucan medical fluid includes: purpose drug
1mg/mL-10mg/mL, mass fraction are the sodium chloride of 0.4%-0.6%, and mass fraction is the glucan of 2%-20%, solvent
For water.
6. as described in claim 1 maintain to carry the method that medicine red blood cell bioactivity improves its drugloading rate simultaneously, feature exists
In in step (1), the volume ratio of the red blood cell and hypotonic glucan medical fluid is 1:(4-20).
7. as described in claim 1 maintain to carry the method that medicine red blood cell bioactivity improves its drugloading rate simultaneously, feature exists
In in step (1), the red blood cell is packed red cells.
8. as described in claim 1 maintain to carry the method that medicine red blood cell bioactivity improves its drugloading rate simultaneously, feature exists
In the preparation method for stating packed red cells at least includes the following steps: animal's whole blood centrifuge separation removes blood plasma and leucocyte,
The packed red cells are obtained after washing.
9. as described in claim 1 maintain to carry the method that medicine red blood cell bioactivity improves its drugloading rate simultaneously, feature exists
In further including one or more in following characteristics:
A. in step (1), the temperature of first time incubation are as follows: 0-10 DEG C；
B. in step (1), the time being incubated for for the first time is 10-60min；
C. it in step (2), is counted on the basis of the high sepage total amount, the constituent of the high sepage includes: that mass fraction is
2.5-5.5% sodium chloride, 1-10mg/ml Sodium Pyruvate and 2-20mg/ml glucose, solvent are water；
D. in step (2), the volume ratio of high sepage and hypotonic glucan medical fluid is 1:(4-20)；
E. in step (2), second of temperature being incubated for are as follows: 30-40 DEG C；
F. in step (2), second of the time being incubated for is 10-60min.
10. the maintenance as described in claim 1-9 is any carries the method that medicine red blood cell bioactivity improves its drugloading rate simultaneously, use
The purposes of medicine red cell preparation is carried in preparation.
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| EP0824213A2 (en) * | 1996-08-15 | 1998-02-18 | Tsukasa Matsumoto | Method for fractionating red blood cells and antibacterial materials or bacterial proliferation inhibitors produced thereby |
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| CN106668840A (en) * | 2017-02-10 | 2017-05-17 | 南通大学 | Insulin controlled-release drug, and preparation method and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113318088A (en) * | 2021-06-16 | 2021-08-31 | 上海交通大学 | Compound medicine and preparation method and application thereof |
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