CN104288185A - Preparation method for polysaccharide nucleic acid medicine composition curing infantile eczema - Google Patents
Preparation method for polysaccharide nucleic acid medicine composition curing infantile eczema Download PDFInfo
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Abstract
The invention relates to a preparation method for polysaccharide nucleic acid medicine composition curing infantile eczema. BCG-polysaccharide nucleic acid and oxymatrine are adopted as medicine active ingredients to prepare an injection sustained release microsphere preparation; the preparation is prepared through a multiple emulsion method by BCG-polysaccharide nucleic acid, oxymatrine, lactide/glycolide copolymer, polyving akohol, Tween 80 and trehalose. The medicine composition can effectively play medicine cure effect in synergism, cures infantile eczema better, and is higher in curing effect, shorter in medicine taking period, and better in medicine taking compliance.
Description
A?preparing?method?of?the?pharmaceutical?composition?of?polysaccharide?nucleic?acid?for?the?treatment?of?pediatric?eczema
Technical field
The invention belongs to medical art, be specifically related to a kind of preparation method for the treatment of the polyoses nucleic acid pharmaceutical composition of infantile eczema.
Background technology
Infantile eczema is a kind of allergic skin disease.Cause the cause of disease of eczema to be complicated, wherein most importantly intolerance factors is, so there is the children's of allergic constitution family history more easily eczema to occur.Main cause is not for tolerate ingestant, inhalation (inhalatio) or contactant or caused by allergy.
The Major Clinical feature of infantile eczema is, erythra pleomorphism, violent scratchiness, often has and oozes out the more symmetrical and easy recurrent exerbation of tendency, distribution, if not in time effectively process make course of disease delay probably cause disease to develop to chronicity.Once suffer from eczema will have a strong impact on the rest of infant, appetite and sleep, infant also easy secondary bacterial infection of repeatedly scratching makes that sb.'s illness took a turn for the worse.
The treatment means of existing infantile eczema mainly removes anaphylactogen; Adopt the symptomatic treatments such as externally used paste; For the infant of acute general, active treatment general disease, remove focus, the main means adopting oral hormone treatment.But existing treatment means, as outer paste etc., is easily scratched by children's, even eats by mistake, there is certain potential safety hazard.Then there is the common adverse effect of hormone therapy in oral hormone treatment, injures greatly children's.Therefore be badly in need of the new treatment means of exploitation and medicine.
Bacill calmette-guerin is cattle type attenuation tulase, and no pathogenicity, has immunogenicity, and replacing tulase primary infection with bacill calmette-guerin inoculation and obtain immunity lungy, is current one of most widely used vaccine in the world.China's researcher has carried out the research of bacill calmette-guerin extract, adopts hot phenol method to remove bacterium protein, then extracts mycelial polysaccharides and nucleic acid and a small amount of protein mixture with alcohol settling and make.
Through the raising of process modification and quality standard, the side reaction of bacill calmette-guerin extract is able to effective reduction, enter " Products in China code ", make BCG polyose nuclear acid injection, clinically for immunomodulating, there is good curative effect at dermatosis such as respiratory tract disease and eczema, urticaria, verruca plana, verruca vulgaris, condyloma acuminatum such as prevention and therapy flu, asthma, allergic rhinitis.The existing part research in this area adopts BCG polyose nuclear acid injection treatment chronic eczema (" Chinese leprosy dermatosis magazine ", Wu Xiaojin etc., the 29th volume the 10th phase, 677-679 page).
Adopt BCG polyose nuclear acid injection treatment infantile eczema to become another selection newly clinically, this kind of therapy safety is good, and curative effect is reliable, effectively can treat infantile eczema.But this kind of therapy needs at least every intramuscular injection in 1 day once, infant poor compliance, and family burden is heavier.
On this basis, inventor have developed a kind of bcg-polysaccharides nucleic acid slow-release injection, effectively can delay drug release, prolong drug action time, increases shot to shot turnaround, improves the compliance of infant.In addition, by adding active constituents of medicine further in injection, drug effect Synergistic can be made, improving therapeutic effect, shorten administration time.
Summary of the invention
The invention reside in and provide a kind of preparation method for the treatment of the polyoses nucleic acid pharmaceutical composition of infantile eczema, described compositions, by increasing active constituents of medicine, makes slow-release injection, can strengthen therapeutic effect, improves the compliance of infant.
The polyoses nucleic acid pharmaceutical composition that the present invention treats infantile eczema is a kind of slow-release microshpere formulation for injection, and its active constituents of medicine is bcg-polysaccharides nucleic acid and oxymatrine.
Wherein, the compositions that active component bcg-polysaccharides nucleic acid is made up of BCG-polysaccharide and bacill calmette-guerin nucleic acid, wherein the mass percentage of polysaccharide is 15% ~ 55%, the mass percentage of said composition amplifying nucleic acid is 45% ~ 85%, preferably, polysaccharide is 40%, and nucleic acid is 60%.
Two kinds of active constituents of medicine in injection of the present invention, the weight proportion of bcg-polysaccharides nucleic acid and oxymatrine is 3:1 ~ 1:3, is preferably 1-3:1 further, is more preferably 1.7:1.Through drug efficacy study of the present invention, the active constituents of medicine of this proportioning can be worked in coordination with and effectively be played curative effect of medication, treats infantile eczema preferably, strengthens therapeutic effect, shortens the medication cycle, improves medication compliance.
The pharmaceutical composition of injection bcg-polysaccharides nucleic acid co-oxidation matrine slow-release microsphere of the present invention, is made up of bcg-polysaccharides nucleic acid, oxymatrine, poly (lactide-co-glycolide), polyvinyl alcohol, Tween 80 and trehalose.Consist of with each component of weight parts:
As a preferred embodiment of the present invention, the pharmaceutical composition of this injection bcg-polysaccharides nucleic acid co-oxidation matrine slow-release microsphere, each component by weight consists of:
The pharmaceutical composition of injection bcg-polysaccharides nucleic acid co-oxidation matrine slow-release microsphere of the present invention adopts following preparation method, and concrete steps comprise:
(1) bcg-polysaccharides nucleic acid of recipe quantity and oxymatrine are dissolved in the PBS buffer solution of the Tween 80 containing recipe quantity trehalose and half recipe quantity, make aqueous phase solution (W1);
(2) hand over fat/co-glycolide dichloromethane to dissolve by third and make oil-phase solution (O), the concentration of oil-phase solution is 0.1-0.5 grams per milliliter; The aqueous phase solution (W1) of above-mentioned drug containing active component is joined in oil-phase solution (O), oil-phase solution (O) is 20: 1 to 2: 1 with the ratio of aqueous phase solution (W1), these two kinds of solution are uniformly mixed at temperature 5-30 DEG C, make colostrum W1/O, preserve at 4-10 DEG C;
(3) by the polyvinyl alcohol of recipe quantity and the Tween 80 of second half recipe quantity, be fully dissolved in water, make aqueous phase solution (W2), the weight concentration of polyvinyl alcohol is 5-20%; Above-mentioned colostrum W1/O is joined aqueous phase solution (W2) stir, the ratio of colostrum W1/O and aqueous phase solution (W2) is 1: 50 to 1: 200, makes emulsion W1/O/W2, preserves at 4-10 DEG C;
(4) emulsion W1/O/W2 is transferred in NaCl aqueous solution, stir, organic solvent is volatilized, then collected by centrifugation microsphere.
Whipping temp in described step (2) is 25 DEG C, and mixing speed is 500-8000 revs/min, and mixing time is 0.02-0.2 hour.
Whipping temp in described step (3) is 15 DEG C, and mixing speed is 100-3000 revs/min, and mixing time is 0.2-0.5 hour.
Whipping temp in described step (4) is 15 DEG C, and mixing speed is 500-3000 revs/min, and mixing time is 0.2-0.5 hour.
The BCG-polysaccharide of composition active constituents of medicine and bacill calmette-guerin nucleic acid, extract by the following method and obtain from bacill calmette-guerin culture:
(1) thalline is collected when bacill calmette-guerin strain inoculation being cultured in the culture medium that applicable mycobacterium grows logarithmic (log) phase;
(2) above-mentioned thalline collected by centrifugation supernatant after fragmentation;
(3) supernatant is after organic solvent extracting, by dialysis or gel chromatography removing organic solvent, obtains BCG-polysaccharide, mixtures of nucleic acids;
(4) BCG-polysaccharide obtained, mixtures of nucleic acids are separated by the column chromatography of anionic exchange medium, loading, washes post with normal saline, collects and washes post liquid;
(5) carry out desalination with desalting column, obtain BCG-polysaccharide;
(6) step (4) is washed post to terminate rear employing concentration is that the sodium chloride solution of 0.1 ~ 2mol/L carries out eluting, collects eluent;
(7) adopt desalting column to carry out desalination the eluent of collection, obtain bacill calmette-guerin nucleic acid;
(8) BCG-polysaccharide prepared and bacill calmette-guerin nucleic acid are mixed in proportion, obtain bcg-polysaccharides nucleic acid mixture.
According to the report that prior art is studied for microsphere, microball preparation is generally framework material with polymer, and described polymer can be biodegradable or biological nondegradable or the biodegradable and nondegradable combination of biology.General employing pharmaceutical acceptable polymer biodegradable but not soluble in water prepares injectable microsphere, include but not limited to polylactide-co-glycolide, polylactic acid, polyglycolic acid, poly-3-hydroxybutyrate ester, polylactic acid-glycollic acid, Poly(D,L-lactide-co-glycolide (PLGA), poly-adjacent ester, polylactone, polyanhydride, polyvinyl alcohol (PVA), Polyethylene Glycol (PEG), Polyhydroxybutyrate-co-hydroxyvalerate copolymer, polypropylene glucosan, polyglycolic acid, polylactic acid-polyglycol, polyglycolic acid-Polyethylene Glycol, gelatin, albumin, mannitol, a kind of in trehalose or two or more mixture etc. wherein.
Adopt above-mentioned can biodegradable polymer time, its molecular weight is preferably in 5,000 ~ 500,000 daltonian scope.Molecular weight is greater than 500, and 000 molecular weight is excessive, enters after in body and is not easy degraded, be likely difficult to realize suitable blood drug level, and can become release time long.Molecular weight is too small lower than 5000 molecular weight, is likely difficult to the object realizing slow release.The degradation rate of polymer expection and the release profiles of inventive compound expection can be depending on the different mixtures of the kind of monomer used, homopolymer used or copolymer and polymer used.
Inventor have passed through the experiment of a large amount of screening, is surprisingly found out that: poly (lactide-co-glycolide) coordinates with polyvinyl alcohol, is more suitable for for the preparation of polyoses nucleic acid sustained release microsphere agents of the present invention than other polymer above-mentioned.Bcg-polysaccharides nucleic acid and oxymatrine are prepared by multi-emulsion method with poly (lactide-co-glycolide) and polyvinyl alcohol, have excellent balling property, and the medicine stability of excellence.
For the preferred poly (lactide-co-glycolide) of the present invention, its molecular weight at 5,000-100, between 000 dalton, preferred molecular weight between 5000-20000, most preferably between 5000-10000.Wherein the polymerization of lactide and Acetic acid, hydroxy-, bimol. cyclic ester is than about between 95: 5-5: 95, is preferably about 40: 60-75: 25, most preferably is about 50: 50.Most preferably the molecular weight of poly (lactide-co-glycolide) is 5000-10000, such as about 5000, about 6000, about 7000, about 8000, about 9000 and about 10,000.The preferred polyvinyl alcohol of the present invention is the polyvinyl alcohol of low polymerization degree, and its molecular weight, at 20,000-150, between 000, is preferably about 20,000, about 25,000, about 30,000.
Microsphere of the present invention may further include other adjuvants, as: a kind of in polyvinylpyrrolidone, Tween 80, sodium carboxymethyl cellulose, dextrin, Polyethylene Glycol, glycerol, glucose and NaGC or two or more mixture etc. wherein.The preferred Tween 80 of the present invention and trehalose.
Through constantly attempting and concentrating on studies, pharmaceutical composition of the present invention, the compatibility of its active component bcg-polysaccharides nucleic acid and oxymatrine achieves the therapeutic effect of Synergistic.Slow releasing preparation can extend dosing interval, promotes infant compliance, improves therapeutic effect.
Accompanying drawing explanation
Accompanying drawing 1 is the In-vitro release curves of the microsphere prepared by embodiment 1, and curve A is the release in vitro of the BCG-polysaccharide in simulation release liquid; Curve B is the release in vitro of the bacill calmette-guerin nucleic acid in simulation release liquid; Curve C is the release in vitro of the oxymatrine in simulation release liquid.
Accompanying drawing 2 is In-vitro release curves of the microsphere prepared by embodiment 2, and curve A is the release in vitro of the BCG-polysaccharide in simulation release liquid; Curve B is the release in vitro of the bacill calmette-guerin nucleic acid in simulation release liquid; Curve C is the release in vitro of the oxymatrine in simulation release liquid.
Detailed description of the invention
The present invention is explained further below in conjunction with embodiment.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.
Embodiment 1
(1) bcg-polysaccharides nucleic acid and oxymatrine are dissolved in the PBS buffer solution that 1 milliliter contains trehalose and half Tween 80, make aqueous phase solution (W1);
(2) hand over fat/co-glycolide to be dissolved in 4 milliliters of dichloromethane by third and make oil-phase solution (O);
(3) above-mentioned aqueous phase solution (W1) is joined in oil-phase solution (O), make these two kinds of solution at temperature 25 DEG C, mixing speed is 8000 revs/min, mixing time is 0.05 hour, be uniformly mixed, make colostrum W1/O, preserve at 4-10 DEG C;
(4) join in the polyvinyl alcohol of 300ml by above-mentioned colostrum W1/O under 1500 revs/min of stirring states, whipping temp is 15 DEG C, and mixing time is 0.2 hour, makes emulsion W1/O/W2, preserves at 4-10 DEG C;
(5) emulsion W1/O/W2 is joined in the 1%NaC1 aqueous solution of 3 times of emulsion volumes, by 1500 revs/min of stirrings, organic solvent is volatilized, then 2000 revs/min, centrifugal 15 minutes, collect microsphere, deposit in refrigerator and cooled and hide.
Obtained microsphere form rounding, even particle size distribution, mean diameter is at 18.2 microns, and drug loading can reach 7.9%, and envelop rate is 47.9%.
Embodiment 2
(1) bcg-polysaccharides nucleic acid and oxymatrine are dissolved in the PBS buffer solution that 2 milliliters contain trehalose and half Tween 80, make aqueous phase solution (W1);
(2) hand over fat/co-glycolide to be dissolved in 6 milliliters of dichloromethane by third and make oil-phase solution (O);
(3) above-mentioned aqueous phase solution (W1) is joined in oil-phase solution (O), make these two kinds of solution at temperature 25 DEG C, mixing speed is 8000 revs/min, mixing time is 0.2 hour, be uniformly mixed, make colostrum W1/O, preserve at 4-10 DEG C;
(4) join in the polyvinyl alcohol of 400ml by above-mentioned colostrum W1/O under 1500 revs/min of stirring states, whipping temp is 15 DEG C, and mixing time is 0.2 hour, makes emulsion W1/O/W2, preserves at 4-10 DEG C;
(5) emulsion W1/O/W2 is joined in the 1%NaC1 aqueous solution of 4 times of emulsion volumes, by 1500 revs/min of stirrings, organic solvent is volatilized, then 2000 revs/min, centrifugal 25 minutes, collect microsphere, deposit in refrigerator and cooled and hide.
Obtained microsphere form rounding, even particle size distribution, mean diameter is at 19.6 microns, and drug loading can reach 7.9%, and envelop rate is 35.6%.
The extracorporeal releasing test of embodiment 3 microsphere
Test specimen: microsphere prepared by the method according to embodiment of the present invention 1-2.
Experimental apparatus: water-bath constant temperature oscillator, centrifuge.
Experiment condition: temperature: 37 ± 0.5 DEG C, rotating speed: 100rpm.
Experimental technique: precision takes laboratory sample and is about 10mg, is placed in the clear bottle of tool lid that volume is 100ml, adds 90ml release medium (Tween-80 of 0.02%), be placed in water-bath constant temperature oscillator, keep certain temperature and rotating speed to sample on time.
Sampling method: essence gets 5ml solution, centrifugal 10min under 3000 turns of conditions, then add the release medium of 5ml, take out liquid HPLC and detect.
Sampling time point (hour): 1,8,16,24,48,3 days, 5 days.
Result of the test: the embodiment of the present invention 1, the microsphere release BCG-polysaccharide preparation of 8 hours prepared by embodiment 2 are respectively 13.8%, 14.6%, and the preparation of 5 days is respectively 94.7%, 95.1%; The bacill calmette-guerin nucleic acid preparation of 8 hours is respectively 12.6%, 11.5%, and the preparation of 5 days is respectively 95.2%, 95.5%; The oxymatrine preparation of 8 hours is respectively 20.3%, 18.5%, and the preparation of 5 days is respectively 93.6%, 89.5%.
Microsphere release test result is see accompanying drawing 1 and accompanying drawing 2.
Embodiment 4 test of pesticide effectiveness
46 routine infants are all from outpatient service, and man 22 example, female 24 example, treats in first 1 month and all do not take Glucocorticoid, do not take antihistamine drug, be not in the mood for hepatorenal disease in 1 week, have erythema, pimple, ooze out, the symptom such as the erythra such as general and violent pruritus.Age is 2 ~ 6 years old, average out to 4.1 years old, and the course of disease is 2d ~ 2 year.Skin injury happening part: Head And Face 2 example, cervical region 3 example, trunk 14 example, extremity 25 example, whole body general 2 example.Clinical diagnosis meets dermatitis and eczema diagnostic criteria.46 routine patients are divided into treatment group and matched group at random, wherein treatment group 25 example, matched group 21 example, two groups of patients equal no difference of science of statistics on sex, age, the course of disease and disease severity, has comparability.
Therapeutic Method: treatment group uses the slow releasing injection of bcg-polysaccharides nucleic acid co-oxidation matrine of the present invention (adopting embodiment 1 prescription), and intramuscular injection, 1ml is each, intramuscular injection 1 time in every 5 days.Matched group adopts commercially available BCG polyose nuclear acid injection (Si Qikang), intramuscular injection, and 1ml is each, and intramuscular injection in every 2 days once.Two groups of courses for the treatment of are 4 weeks, all further consultation in the 10th, 28 day after treatment.
Curative effect determinate standard: before the treatment, record erythema, pimple, erosion respectively on the 10th, 28 day after treatment, ooze out, infiltrate or lichenization, keratinization desquamation etc. and pruritus degree, each index is undertaken by 4 grades of point systems when assessment.Pruritis: 0=is without gargalesthesia, and pruritus that 1=is slight, 2=moderate pruritus but can stand, 3=severe pruritus is impatient at.Erythra sign: 0=without, 1=is slight, 2=moderate, 3=severe, according to before treatment, the total mark of rear corresponding follow up time calculates symptom integral decline index with treatment, decline index is divided into recovery from illness, effective, effective and invalid 4 grades of assessment curative effects, and computing formula is as follows: integration × 100% before symptom integral decline index=(before treatment the rear integration of integration-treatment)/treatment.Recovery from illness: symptom integral decline index is more than or equal to 90%; Effective: symptom integral decline index is more than or equal to 60%, but is less than 90%; Effective: symptom integral decline index is more than or equal to 20%, but is less than 60%; Invalid: symptom integral decline index is less than 20%.Total obvious effective rate is recovery from illness and effective patient's percent sum.
Clinical efficacy is in table 1.Treatment group starts the shortest 5d of effective time, the longest 10d, and matched group starts the shortest 9d of effective time, the longest 14d.Two groups of comparitive study during 10d, the total obvious effective rate 53% for the treatment of group, there were significant differences (χ 2=6.215, P<0.05) for the total obvious effective rate of matched group 40%, two groups of total obvious effective rates.Two groups of comparitive study during 28d, the total obvious effective rate 92% for the treatment of group, the total obvious effective rate of matched group 61%, two groups of total obvious effective rate differences have significant (χ 2=11.616, P<0.01).Wherein during 28d check, matched group has 3 people lost to follow-up, and treatment group all adheres to treatment.
Table 1 Clinical efficacy comparison
Embodiment 5 test of pesticide effectiveness
Adopt rat subacute eczema model, bcg-polysaccharides nucleic acid and oxymatrine proportioning are studied.Modeling method is as follows: get SD rat, about 220g, and pentobarbital sodium is anaesthetized, and at rat back labelling, area 1 × 1cm, removes Mus hair, sticks with paste with 6% sodium sulfide mixing starch, removes formation shave hair-fields after 5 minutes with water.Shaving the DNFB acetone sensitization that hair-fields smears 7%, matched group such as to smear at the water gaging, first after sensitization, administration group gives drug dose and be 30mg/kg (lumbar injection), the water gagings such as model group and matched group lumbar injection every day, repeat sensitization once weekly, continue surrounding.Shave hair-fields according to eczema area and Severity Index (EASI) to each treated animal to mark from erythema, hard swollen and exfoliation three aspects respectively, the region sign atopic dermatitis point system of the severity score reference berth-Jones of every aspect performance, 0=without, 1=is light, in 2=, 3=is heavy, and each symptom score can remember half score value.
The percentage by weight of each administration group bcg-polysaccharides nucleic acid and oxymatrine sees table
| ? | Bcg-polysaccharides nucleic acid | Oxymatrine |
| Administration group 1 | 100％ | 0％ |
| Administration group 2 | 0％ | 100％ |
| Administration group 3 | 50％ | 50％ |
| Administration group 4 | 62.5％ | 37.5％ |
| Administration group 5 | 66.6％ | 33.4％ |
| Administration group 6 | 75％ | 25％ |
Result
(1) administration group is on the impact (n=8) of rat Eczema Model skin lesion
| Scoring | Erythema | Edema | Exfoliation |
| Matched group | 0 | 0 | 0 |
| Model group | 2.98±0.34 | 2.36±0.41 | 1.79±0.38 |
| Administration group 1 | 1.21±0.16 | 1.17±0.22 | 0.86±0.21 |
| Administration group 2 | 1.87±0.18 | 1.72±0.31 | 1.23±0.28 |
| Administration group 3 | 0.74±0.12 | 0.57±0.06 | 0.71±0.09 |
| Administration group 4 | 0.21±0.05 | 0.21±0.03 | 0.51±0.06 |
| Administration group 5 | 0.34±0.08 | 0.38±0.07 | 0.64±0.08 |
| Administration group 6 | 0.69±0.09 | 0.51±0.05 | 0.69±0.08 |
(2) administration group is on the impact of horn cell (KC) and lymphocyte (LYM) apoptosis and IL-4 in rat Eczema Model
In administration after 2 weeks, put to death the partial rat in each group respectively, be separated rat eczema skin lesion piece of tissue, paraffin section after paraformaldehyde is fixing, adopt ImmunohistochemistryMethods Methods to detect the apoptosis of KC and LYM and the content of IL-4 in section respectively, concrete outcome is as follows:
| ? | KC apoptosis rate (%) | LYM apoptosis rate (%) | The IOD value of IL-4 |
| Matched group | - | - | 79±6 |
| Model group | 9.3±0.9 | 10.3±1.1 | 173±8 |
| Administration group 1 | 29.5±1.6 | 28.7±1.8 | 121±7 |
| Administration group 2 | 18.2±1.2 | 15.4±1.4 | 169±9 |
| Administration group 3 | 41.6±2.3 | 38.4±0.6 | 104±6 |
| Administration group 4 | 58.9±1.8 | 56.7±1.1 | 82±5 |
| Administration group 5 | 51.4±1.6 | 48.7±0.7 | 91±4 |
| Administration group 6 | 44.7±0.9 | 41.2±0.8 | 113±8 |
Content of the present invention merely illustrates some claimed specific embodiments; one of them or more described technical characteristic can be combined with arbitrary one or more technical scheme in technical scheme; these technical schemes obtained through combination also in the application's protection domain, just as these technical schemes obtained through combination in the disclosure of invention concrete record.
Claims (7)
1. treat a preparation method for the polyoses nucleic acid pharmaceutical composition of infantile eczema, it is characterized in that concrete steps comprise:
(1) bcg-polysaccharides nucleic acid of recipe quantity and oxymatrine are dissolved in the PBS buffer solution of the Tween 80 containing recipe quantity trehalose and half recipe quantity, make aqueous phase solution (W1);
(2) hand over fat/co-glycolide dichloromethane to dissolve by third and make oil-phase solution (O), the concentration of oil-phase solution is 0.1-0.5 grams per milliliter; The aqueous phase solution (W1) of above-mentioned drug containing active component is joined in oil-phase solution (O), oil-phase solution (O) is 20: 1 to 2: 1 with the ratio of aqueous phase solution (W1), these two kinds of solution are uniformly mixed at temperature 5-30 DEG C, make colostrum W1/O, preserve at 4-10 DEG C;
(3) by the polyvinyl alcohol of recipe quantity and the Tween 80 of second half recipe quantity, be fully dissolved in water, make aqueous phase solution (W2), the weight concentration of polyvinyl alcohol is 5-20%; Above-mentioned colostrum W1/O is joined aqueous phase solution (W2) stir, the ratio of colostrum W1/O and aqueous phase solution (W2) is 1: 50 to 1: 200, makes emulsion W1/O/W2, preserves at 4-10 DEG C;
(4) emulsion W1/O/W2 is transferred in NaCl aqueous solution, stir, organic solvent is volatilized, then collected by centrifugation microsphere;
The prescription of described compositions is:
Bcg-polysaccharides nucleic acid 3-1 part
Oxymatrine 1-3 part
Poly (lactide-co-glycolide) 0.5 ~ 12 part
Polyvinyl alcohol 0.5 ~ 6 part
Tween 80 0.1 ~ 6 part
Trehalose 0.05 ~ 1.2 part.
2. the preparation method of pharmaceutical composition according to claim 1, is characterized in that in described method, and the whipping temp in step (2) is 25 DEG C, and mixing speed is 500-8000 revs/min, and mixing time is 0.02-0.2 hour.
3. the preparation method of pharmaceutical composition according to claim 1, is characterized in that in described method, and the whipping temp in step (3) is 15 DEG C, and mixing speed is 100-3000 revs/min, and mixing time is 0.2-0.5 hour.
4. the preparation method of pharmaceutical composition according to claim 1, is characterized in that in described method, and the whipping temp in step (4) is 15 DEG C, and mixing speed is 500-3000 revs/min, and mixing time is 0.2-0.5 hour.
5. the preparation method of pharmaceutical composition according to claim 1, it is characterized in that its molecular weight of described poly (lactide-co-glycolide) is at 5,000-100, between 000 dalton, preferred molecular weight between 5000-20000, most preferably between 5000-10000.
6. the preparation method of pharmaceutical composition according to claim 1, it is characterized in that described poly (lactide-co-glycolide), the polymerization of its lactide and Acetic acid, hydroxy-, bimol. cyclic ester, than about between 95: 5-5: 95, is preferably about 40: 60-75: 25, most preferably is about 50: 50.
7. the preparation method of pharmaceutical composition according to claim 1, is characterized in that described polyvinyl alcohol is the polyvinyl alcohol of low polymerization degree, and its molecular weight, at 20,000-150, between 000, is preferably about 20,000, about 25,000, about 30,000.
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| CN113230036A (en) * | 2021-04-15 | 2021-08-10 | 杭州千岛湖天鑫有限公司 | Sea-buckthorn oil slow-release diaper |
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| CN102106828A (en) * | 2011-02-25 | 2011-06-29 | 深圳翰宇药业股份有限公司 | Thymalfasin sustained-release microsphere preparation and preparation method thereof |
| CN103565779A (en) * | 2013-09-11 | 2014-02-12 | 中国药科大学 | Oxymatrine biological adhering sustained release preparation and preparation method thereof |
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| CN113230036A (en) * | 2021-04-15 | 2021-08-10 | 杭州千岛湖天鑫有限公司 | Sea-buckthorn oil slow-release diaper |
| CN113230036B (en) * | 2021-04-15 | 2022-10-14 | 杭州千岛湖天鑫有限公司 | Sea-buckthorn oil slow-release diaper |
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