CN106038492B - A kind of preparation method preparing sustained release leuprorelin acetate microballoon - Google Patents
A kind of preparation method preparing sustained release leuprorelin acetate microballoon Download PDFInfo
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Abstract
The purpose of the present invention is to provide a kind of preparation method of sustained release leuprorelin acetate microballoon, the step of preparation method, is：(1) leuprorelin acetate is dissolved in water for injection, forms leuprorelin acetate aqueous solution for injection, a concentration of 1g/ml 2.5g/ml of leuprorelin acetate；(2) PLA is dissolved in dichloromethane, formation PLA dichloromethane solutions, a concentration of 280mg/ml 350mg/ml of PLA in the PLA dichloromethane solutions；(3) the leuprorelin acetate aqueous solution for injection and PLA dichloromethane solutions of the step (1) and the step (2) are mixed, is ultrasonically formed colostrum；(4) step (3) described colostrum is added in PVA solution, stirring forms emulsion；(5) emulsion stirs, filtering, mannitol solution is added into microballoon wet product, freeze-drying is to get sustained release leuprorelin acetate microballoon.
Description
Technical field
The present invention relates to a kind of preparation methods of the lyophilized preparation of sustained release leuprorelin acetate microballoon.
Background technology
Leuprorelin acetate is the LH-RH derivatives of high activity, resistance to protease and to LH-RH receptors
Affinity ratio LH-RH is strong, can effectively inhibit the function of hypophysis-Sexual gland system, and clinic is mainly used for prostate cancer and intrauterine
The treatment of endometriosis and premenopausal breast cancer patients.Since the treatment of prostate cancer and endometrium disease and premenopausal breast cancer patients is usual
Need the several months even 1 year or more to 1 year, so common Lupron Injection and sustained release microsphere agents meeting in 1 month
Brought to patient's administration greatly constant, and the research listing of longer time sustained release preparation being ground as Lupron Injection
Hair trend.
3 months leuprorelin acetate PLA sustained-release micro-spheres (trade name " LUPRON DEPOT-PED ") in Japanese force field are to adopt
With biodegradable polymers be framework material packaging medicine formed for injection administration microball preparation, can be at 3 months
It is interior that drug is discharged to maintain effective blood drug concentration with given pace, reach raising therapeutic effect, reduce administration number of times, reduces medicine
Object toxic side effect increases patient compliance, mitigates sufferer pain and medical treatment burden.
In addition to this document also having discloses 3 months schemes of leuprorelin acetate microballoon,《Sustained-release leuprorelin microspheres
Research》(Feng Lan, waits the research of sustained-release leuprorelin microspheres, Chinese Journal of New Drugs and Clinical Remedies, in October, 2004, volume 23
10 phases,
5% sodium chloride 680-683) is added in water phase makes w/o/w emulsions more stablize, and 3 months sustained releases of preparation are micro-
117 μm of ball average grain diameter increases a kind of addition of auxiliary material sodium chloride, and the grain size of microballoon compared with commercialized product prescription
It is significantly increased；Industrialized difficulty can be significantly increased in the auxiliary material newly increased in the document, and the increasing of microspherulite diameter can increase
Pain degree when clinical injection.
500mg leuprorelin acetates were dissolved in distilled water by Hiroaki Okada in 1994, and 4g PLA are dissolved in
In 7.5ml dichloromethane, water phase and oil phase are prepared respectively, then prepare leuprorelin acetate microballoon, load medicine is disclosed in research
Influence of the molecular weight of amount and PLA for 3 months leuprorelin acetate PLA sustained-release micro-spheres release performances, finds PLGA and PLA phases
Than the latter's slow-release time is longer, and it is more suitable using PLA for 3 months sustained-release micro-spheres to illustrate；Drugloading rate raising can increase burst release,
Prepared by the document can be 3 months with sustained release by prescription and the preferred microballoon of technique, but can all be generated at the 8th week primary
Serious burst release (H.Okada, Y.Doken, Y.Ogawa, H.Toguchi, Preparation of three-month depot
Injectable microspheres of leuprorelin acetate using biodegradable polymers,
Pharm.Res.11(1994)1143-1147)。
550mg leuprorelin acetates were dissolved in distilled water by Hiroaki Okada in 1997, and 4g PLA are dissolved in
In 7.5ml dichloromethane, water phase and oil phase are prepared respectively, then prepares leuprorelin acetate microballoon, and it is auspicious to increase acetic acid bright third
Concentration of the woods in water phase, and PLA is remained unchanged in the concentration of oil phase, the 3 months leuprorelin acetate PLA disclosed in research are slow
Primary serious burst release (Okada H.One and three can all be led to the problem of at the 8th week by releasing microballoon and remaining
release injectable microspheres of LH-RH susperagonist leuprorelin acetate
[J] .Adv Drug Deliv Rev, 1997,28 (1)：43-70).
PLA is polylactic acid (polylactic acid), is the polymer that main polymerizable raw material obtains with lactic acid, is a kind of
Degradable functional polymer organic compound.
PLGA is Poly(D,L-lactide-co-glycolide (poly (lactic-co-glycolic acid)), by two kinds of lists
Body-lactic acid and hydroxyacetic acid are polymerized at random, are a kind of degradable functional polymer organic compounds.
Invention content
The purpose of the present invention is to provide a kind of new leuprorelin acetate PLA sustained-release micro-spheres, which can press down
Leuprorelin acetate initial transition release processed, reduces the rate of release in 72 hours after injecting, and release is more steady, burst release drop
It is low.
The purpose of the present invention is to provide a kind of new leuprorelin acetate PLA sustained-release micro-spheres preparation methods, party's legal systems
Release after the standby leuprorelin acetate PLA sustained-release micro-spheres injection obtained in 72 hours is more steady, and burst release reduces.
In order to obtain the more stable leuprorelin acetate PLA sustained-release micro-spheres of release, the present inventor furthers investigate,
It was found that in preparation process the leuprorelin acetate concentration of leuprorelin acetate aqueous solution for injection with burst release there is being associated with,
In certain concentration range, the microballoon release being prepared is steady, and the burst release in 72 hours is greatly reduced；It is below or above
The microballoon release of the specific concentration, acquisition is steady after 72 hours, but will appear significant phenomenon of burst release in 72 hours.
Further, in order to obtain the microballoon for meeting quality standard, the present inventor also has adjusted PLA dichloros simultaneously
PLA concentration in methane has been surprisingly found that the freshly prepared microballoon being adapted with leuprorelin acetate concentration can not only meet quality mark
Standard, and the D90 grain sizes of microballoon are lowered 40% or so.The grain size of microballoon reduces the pain that can reduce injection, is more applicable in
In clinic.
A kind of leuprorelin acetate method for preparing microsphere, it is characterised in that the step of preparation method is：
(1) leuprorelin acetate is dissolved in water for injection, forms leuprorelin acetate aqueous solution for injection；
(2) PLA is dissolved in dichloromethane, forms PLA dichloromethane solutions；
(3) by the leuprorelin acetate aqueous solution for injection and PLA dichloromethane of the step (1) and the step (2)
Solution mixes, and is ultrasonically formed colostrum；
(4) step (3) described colostrum is added in PVA solution, stirring forms emulsion；
(5) emulsion stirs, and filtering obtains microballoon wet product, and mannitol solution is added into microballoon wet product, is lyophilized, and both obtains acetic acid
Leuprorelin microballoon.
A concentration of 1g/ml- of leuprorelin acetate of leuprorelin acetate aqueous solution for injection described in the step (1)
2.5g/ml。
A concentration of 1.5g/ml of leuprorelin acetate of leuprorelin acetate aqueous solution for injection described in the step (1).
A concentration of 280mg/ml-350mg/ml of PLA in PLA dichloromethane solutions described in the step (2).
A concentration of 320mg/ml of PLA in PLA dichloromethane solutions described in the step (2).
A kind of preparation method of leuprorelin acetate microballoon, prepares in accordance with the following steps：
(1) leuprorelin acetate is dissolved in water for injection, forms leuprorelin acetate aqueous solution for injection；
(2) PLA is dissolved in dichloromethane, forms PLA dichloromethane solutions；
(3) by the leuprorelin acetate aqueous solution for injection and PLA dichloromethane of the step (1) and the step (2)
Solution mixes, and is ultrasonically formed colostrum (o/w)；
(4) step (3) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution, stirring is formed
Emulsion (w/o/w)；
(5) emulsion (w/o/w) stirs, and forms microballoon；
(6) it filters, at microballoon wet product；
(7) mannitol solution is added into microballoon wet product, is lyophilized, both obtains leuprorelin acetate microballoon.
The weight average molecular weight of PLA described in the step (2) is 12000-18000.
Leuprorelin acetate microballoon described in the step (7) in vivo can sustained release 8 weeks or more.
Leuprorelin acetate microballoon described in the step (7) in vivo can sustained release 12 weeks or so.
A kind of preparation method of sustained release leuprorelin acetate microballoon, step are：
(1) 225mg leuprorelin acetates are dissolved in 150ul waters for injection, it is 1.5g/ml's to form leuprolide concentrations
Leuprorelin aqueous solution for injection；
(2) 2g PLA are dissolved in 6.26ml dichloromethane, the PLA dichloromethane for forming a concentration of 320mg/ml of PLA is molten
Liquid；
(3) above two solution is mixed, is ultrasonically formed colostrum (o/w)；
(4) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution of 2L 0.5%, stirring forms multiple
Newborn (w/o/w)；
(5) emulsion (w/o/w) stirs, and removes dichloromethane, forms microballoon；
(6) it filters, removes PVA solution, form microballoon wet product；
(7) mannitol solution is added into microballoon wet product, is lyophilized, both obtains leuprorelin acetate microballoon.
The weight average molecular weight of PLA described in the step (2) is 12000-18000.
Three months sustained release leuprorelin acetate microballoons refer to the leuprorelin acetate microballoon can continue in vivo
Release 3 months.
Description of the drawings
Fig. 1：Plasma concentration curves of the G1 and G2 in 72 hours
Fig. 2：Plasma concentration curves of the G1 and G3 in 12 weeks
Fig. 3：The plasma concentration curve of S1, S2, S4, S5 and S6 in 72 hours
Fig. 4：The plasma concentration curve of S2, S4, S5 in 72 hours
Fig. 5：Influence curve figure of the PLA concentration to grain size
Fig. 6：Influence curve figure of the PLA concentration to drugloading rate
Specific embodiment
Below will by specific example, the present invention will be further described, it is to be noted that following embodiment cannot
Constitute any restrictions to invention.
Embodiment 1
(1) 225mg leuprorelin acetates are dissolved in 150ul waters for injection, it is 1.5g/ml's to form leuprolide concentrations
Leuprorelin aqueous solution for injection；
(2) 2g PLA are dissolved in 6.26ml dichloromethane, the PLA dichloromethane for forming a concentration of 320mg/ml of PLA is molten
Liquid；
(3) above two solution is mixed, is ultrasonically formed colostrum (o/w)；
(4) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution of 2L 0.5%, stirring forms multiple
Newborn (w/o/w)；
(5) emulsion (w/o/w) stirs, and removes dichloromethane, forms microballoon；
(6) it filters, removes PVA solution, form microballoon wet product；
(7) suitable mannitol solution is added, freeze-drying both obtains leuprorelin acetate microballoon.
Embodiment 2
(1) 225mg leuprorelin acetates are dissolved in water for injection, form Leuprorelin aqueous solution for injection；
(2) 2g PLA are dissolved in 6.26ml dichloromethane, the PLA dichloromethane for forming a concentration of 320mg/ml of PLA is molten
Liquid；
(3) above two solution is mixed, is ultrasonically formed colostrum (o/w)；
(4) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution of 2L 0.5%, stirring forms multiple
Newborn (w/o/w)；
(5) emulsion (w/o/w) stirs, and removes dichloromethane, forms microballoon；
(6) it filters, removes PVA solution, form microballoon wet product；
(7) suitable mannitol solution is added, freeze-drying both obtains leuprorelin acetate microballoon.
The leuprolide concentrations in Leuprorelin aqueous solution for injection in the step (1) be adjusted to 0.9g/ml,
1.0g/ml, 1.3g/ml, 1.5g/ml, 2.0g/ml, 2.5g/ml, 2.7g/ml and method test blood medicine according to embodiment 2 is dense
Degree, is divided into S1 groups, S2 groups, S3 groups, S4 groups, S5 and S6 groups, blood concentration the results are shown in Table 1, with S1 groups, S2 groups, S4 groups, S5 groups
Fig. 1 is drawn with the time of S6 groups and blood concentration；S2 groups, S3 groups, the time of S4 groups and S5 and blood concentration draw Fig. 2.
Table 1：The blood of Leuprorelin after 0.35mg/kg Leuprorelin preparations S1-S6 is given in male beagle dogs' hypodermic injection
Concentration (ng/mL)
Conclusion：From the result of Fig. 1, although prepared by the Leuprorelin microballoon of S1 groups, S2 groups, S4 groups, S5 and S6 groups
All offer medicine 225mg, but since the leuprolide concentrations in Leuprorelin aqueous solution for injection are different so that is be prepared is bright
There are larger differences for the release characteristics of third Rayleigh microballoon.Wherein S1 groups and S6 group initial release amplitudes of variation is big；And S2 groups, S4
The release of the Leuprorelin microballoon of group, S5 groups is more steady, therefore the leuprolide concentrations in Leuprorelin aqueous solution for injection
For more stable Leuprorelin microballoon can be released when 1.0g/ml to 1.5g/ml.
The data that the Leuprorelin release blood concentration of analysis S2 groups-S5 groups changes over time, Leuprorelin water for injection
When leuprolide concentrations in solution are less than 1.5g/ml, as leuprolide concentrations rise, initial release tends to be steady；And work as
When leuprolide concentrations in Leuprorelin aqueous solution for injection are higher than 1.5g/ml, initial release is accelerated to increase, by diagnostic cast
Analysis is fitted, inventor thinks to be prepared when the leuprolide concentrations in Leuprorelin aqueous solution for injection are 1.5g/ml
Leuprorelin microballoon release it is the most steady.
Embodiment 3
Two groups of male beagle dogs：G1 and G2, after 0.35mg/kg Leuprorelins are given in every group of 4 hypodermic injections respectively；In not
Blood serum sample is acquired with time point, is surveyed respectively using liquid chromatography tandem mass spectrometry after solid phase extraction and organic solvent processing
Determine the concentration of Leuprorelin in G1 and G2 group serum.The quantification range of Leuprorelin is 0.100-50.0ng/mL and 0.0200-
10.0ng/mL。
The leuprorelin acetate of male beagle dogs' injection of G1 groups is prepared according to the method described in the embodiment of the present invention 1；
The leuprorelin acetate of male beagle dogs' injection of G2 groups is 3 months leuprorelin acetate PLA sustained-release micro-spheres (commodity
Name " LUPRON DEPOT-PED ").
4 male beagle dogs number of G1 groups is respectively G1M01, G1M02, G1M03, G1M04；
4 male beagle dogs number of G2 groups is respectively G2M01, G2M02, G2M03, G2M04；
The blood concentration of each group Leuprorelin is shown in Table 2 and table 3.
Table 2：It is auspicious by after test preparation (G1) bright third that 0.35mg/kg Leuprorelins are given in male beagle dogs (n=4) hypodermic injection
The blood concentration (ng/mL) of woods
Table 3：Male beagle dogs (n=4) are subcutaneously injected that give after 0.35mg/kg Leuprorelins reference preparation (G2) bright third auspicious
The blood concentration (ng/mL) of woods
*BLQ：Less than lower limit of quantitation (0.100ng/mL)
ND：It can not detect, participate in calculating with zero.
Conclusion：In terms of Fig. 3 data drawn from table 2 and with table 3, the blood concentration of G2 groups after injection 0.5 hour,
25ng/ml it has been above at 1 hour, 2 hours, 3 hours, 1 hour when reaches highest 44.4ng/ml, after the 4th hour
Less than the blood concentration peak of G1 groups, and between 4 hours to 24 hours downward trend clearly, within 20 hours from
16.9ng/ml dropping to 0.708ng/ml.The unexpected release of G2 groups and release decrease amplitude are all very big.And the blood concentration of G1 groups
Raising and lowering trend in 72 hours is all than shallower, and compared with G2 groups, the decline of initial release and blood concentration is all non-
Often mitigate.Therefore the release of Leuprorelin microballoon that G1 groups are injected is more steady, and initial release properties are better than having listed
“LUPRON DEPOT-PED”。
Stability data of 4 microballoon of embodiment under 30 DEG C of acceleration environments
At 30 DEG C ± 2 DEG C, accelerated stability investigation is carried out under the conditions of RH65% ± 5%, it is 12 months to investigate the time；
It investigates sample and is divided into G1 combination G2 groups, wherein the leuprorelin acetate of G1 groups is according to described in the embodiment of the present invention 1
It is prepared by method；The leuprorelin acetate of G2 groups is 3 months leuprorelin acetate PLA sustained-release micro-spheres (trade name " LUPRON
DEPOT-PED”)。
Investigation project is G1 groups and G2 groups in the related of 0th month and 12nd month leuprorelin acetate PLA sustained-release micro-spheres
Substance, the present invention detect related substance with HPLC methods.
Experimental result data is shown in table 4 below.
Table 4：The stability data of G1 groups and G2 groups microballoon under 30 DEG C of acceleration environments
Conclusion：It is investigated by carrying out 12 months accelerated stabilities at 30 DEG C ± 2 DEG C, under the conditions of RH65% ± 5%, G1 groups
Content is 99.5%, and the content of G2 groups is 98.7%, and the stability of leuprorelin acetate PLA sustained-release micro-spheres of the invention is good
It is good.
The dichloromethane of 5 microballoon of embodiment remains
The leuprorelin acetate PLA sustained-release micro-spheres about 50mg of G1 groups is taken, accurately weighed, in top set empty bottle, precision measures 2ml
Dimethyl sulphoxide solution, which shakes up, to be made it dissolve, sealing, as test solution；Methylene chloride q separately is taken, it is accurately weighed, add two
Methyl sulfoxide, which quantitatively dilutes, is made the solution that every 1ml contains 0.825 μ g, precision measurement 2ml, in top set empty bottle, sealing, as a contrast
Product solution.(four general rules 0521 of Chinese Pharmacopoeia version in 2015) are measured in accordance with the law, with the poly- silica of -94% dimethyl of 6% cyanogen propyl phenyl
Alkane (or polarity is close) is fixer, ECD detectors, 40 DEG C of column temperature initial temperature, maintenance 6 minutes, with the rate of 100 DEG C/min
200 DEG C are risen to, is maintained 2 minutes；250 DEG C of injector temperature；300 DEG C of detector temperature；2: 1 head space equilibrium temperature 90 of split ratio
℃；Equilibration time 20 minutes takes test solution and reference substance solution to distinguish headspace sampling, records chromatogram, by external standard method with
Calculated by peak area.Wherein the leuprorelin acetate of G1 groups is prepared according to the method described in the embodiment of the present invention 1.
3 months leuprorelin acetate PLA sustained-release micro-spheres (trade name " LUPRON DEPOT-PED ") in Japanese force field are open
Dichloromethane residue criterion be not higher than 0.06%.
The dichloromethane residual of G1 group leuprorelin acetate PLA sustained-release micro-spheres prepared by 1 method according to embodiments of the present invention
Amount is 0.0010%, and the method described in inventor according to embodiments of the present invention 1 is prepared for multiple batches of leuprorelin acetate
PLA sustained-release micro-spheres dichloromethane remains, and the leuprorelin acetate PLA sustained-release micro-spheres dichloromethane residue criterions of the present invention are set
To be not higher than 0.0015%, well below margin of safety as defined in Yuan Yan producers.
Embodiment 6
(1) 225mg leuprorelin acetates are dissolved in 150ul waters for injection, it is 1.5g/ml's to form leuprolide concentrations
Leuprorelin aqueous solution for injection；
(2) 2g PLA are dissolved in dichloromethane, form PLA dichloromethane solutions；
(3) above two solution is mixed, is ultrasonically formed colostrum (o/w)；
(4) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution of 2L 0.5%, stirring forms multiple
Newborn (w/o/w)；
(5) emulsion (w/o/w) stirs, and removes dichloromethane, forms microballoon；
(6) it filters, removes PVA solution, form microballoon wet product；
(7) suitable mannitol solution is added, freeze-drying both obtains leuprorelin acetate microballoon.
The PLA concentration in PLA dichloromethane solutions in the step (1) be adjusted to 250mg/ml, 280mg/ml,
300mg/ml, 320mg/ml, 350mg/ml, 360mg/ml, 400mg/ml, be prepared Leuprorelin microballoon S7, S8, S9,
S10, S11 and S12 take 3 months leuprorelin acetate PLA sustained-release micro-spheres (quotient in the military field of the Leuprorelin microballoon of S7-12 and Japan
The name of an article " LUPRON DEPOT-PED "), measure its grain size and drugloading rate.
Particle size determination：Sample about 30mg is taken, 10ml purified waters are added, 3~8min of ultrasound makes to be uniformly dispersed, into particle size analyzer
Sample-adding is measured and is recorded and the results are shown in Table 5 until obscurity reaches 5%~15%.
Table 5：Influence of the PLA concentration to grain size
Conclusion：The measurement result of grain size and drugloading rate is shown in Table lattice 5.It is bright as a concentration of 360mg/ml and 400mg/ml of PLA
The D90 grain sizes of third Rayleigh microballoon are 36.492 μm and 40.855 μm, and grain size is more than 30.000 μm, and when PLA concentration is not higher than
When 350mg/ml, the D90 grain sizes of Leuprorelin microballoon are about 30 μm or are less than 30 μm.It is bright as a concentration of 250mg/ml of PLA
When the drugloading rate of third Rayleigh microballoon is a concentration of 280mg/ml of 5.3%, PLA, drugloading rate 6.5%, PLA concentration is higher than 280mg/
When ml, drugloading rate is not less than 7.0%.
Microspherulite diameter conference easily causes injection pain, and the low injection volume that can increase of drugloading rate, it is therefore desirable to select grain size
The Leuprorelin microballoon all more balanced with drugloading rate, according to the PLA concentration of table 5 to the influence data of grain size and drugloading rate,
The preferred PLA concentration ranges of inventor are 280mg/ml-350mg/ml, a concentration of 320mg/ml of best PLA.
When PLA concentration ranges are 280mg/ml-350mg/ml, the D90 grain sizes for the Leuprorelin microballoon that the present invention obtains
Less than 3 months leuprorelin acetate PLA sustained-release micro-spheres (trade name " LUPRON DEPOT-PED ") in Japanese military field；PLA concentration
For 320mg/ml when, only 21.106 μm of the D90 grain sizes for the Leuprorelin microballoon that the present invention obtains, the original than having listed grinds Japanese force
The Leuprorelin microballoon D90 grain sizes in field reduce 34.0%, while the original that drugloading rate has slightly above listed grinds the bright of Japanese military field
Third Rayleigh microballoon drugloading rate.
Embodiment 7
Two groups of male beagle dogs：G1 and G3, after 0.35mg/kg Leuprorelins are given in every group of 4 hypodermic injections respectively；In not
Blood serum sample is acquired with time point, is surveyed respectively using liquid chromatography tandem mass spectrometry after solid phase extraction and organic solvent processing
Determine the concentration of Leuprorelin in G1 and G3 group serum.The quantification range of Leuprorelin is 0.100-50.0ng/mL and 0.0200-
10.0ng/mL。
The leuprorelin acetate of male beagle dogs' injection of G1 groups is prepared according to the method described in the embodiment of the present invention 1；
The leuprorelin acetate of male beagle dogs' injection of G3 groups is 3 months leuprorelin acetate PLA sustained-release micro-spheres, according to
Okada H.One and three release injectable microspheres of LH-RH susperagonist
Leuprorelin acetate [J] .Adv Drug Deliv Rev, 1997,28 (1)：Prepared by 43-70 the methods, use
PLA molecular weight is 14100.It the results are shown in Table 6.
Table 6：The blood of Leuprorelin after 0.35mg/kg Leuprorelin preparations G1 and G3 is given in male beagle dogs' hypodermic injection
Concentration (ng/mL)
Conclusion：Fig. 2 data drawn from table 3, the blood concentration of G1 groups and G3 groups after injection the 1st week, the 2nd week and
4th week all relatively more steady, but the blood concentration of G3 groups occurred a unexpected rising at the 8th week, at next 10th week
Restore again to stable state with 12 weeks blood concentrations.And raising and lowering trend of the blood concentration of G1 groups in 12 weeks all compares
Shallower, compared with G3 groups, the overall trend of Leuprorelin release and blood concentration is all relatively more steady.Therefore G1 groups are injected
The release of Leuprorelin microballoon is more steady, and release characteristics are better than Leuprorelin microballoon disclosed in Okada H documents.
Embodiment 8
(1) 225mg leuprorelin acetates are dissolved in water for injection, form bright third that leuprolide concentrations are 2.5g/ml
Rayleigh aqueous solution for injection；
(2) 2g PLA are dissolved in 7.14ml dichloromethane, the PLA dichloromethane for forming a concentration of 280mg/ml of PLA is molten
Liquid；
(3) above two solution is mixed, is ultrasonically formed colostrum (o/w)；
(4) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution of 2L 0.5%, stirring forms multiple
Newborn (w/o/w)；
(5) emulsion (w/o/w) stirs, and removes dichloromethane, forms microballoon；
(6) it filters, removes PVA solution, form microballoon wet product；
(7) suitable mannitol solution is added, freeze-drying both obtains leuprorelin acetate microballoon.
Embodiment 9
(1) 225mg leuprorelin acetates are dissolved in water for injection, form bright third that leuprolide concentrations are 2.0g/ml
Rayleigh aqueous solution for injection；
(2) 2g PLA are dissolved in 6.26ml dichloromethane, the PLA dichloromethane for forming a concentration of 320mg/ml of PLA is molten
Liquid；
(3) above two solution is mixed, is ultrasonically formed colostrum (o/w)；
(4) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution of 2L 0.5%, stirring forms multiple
Newborn (w/o/w)；
(5) emulsion (w/o/w) stirs, and removes dichloromethane, forms microballoon；
(6) it filters, removes PVA solution, form microballoon wet product；
(7) suitable mannitol solution is added, freeze-drying both obtains leuprorelin acetate microballoon.
Embodiment 10
(1) 225mg leuprorelin acetates are dissolved in water for injection, form bright third that leuprolide concentrations are 1.0g/ml
Rayleigh aqueous solution for injection；
(2) 2g PLA are dissolved in 6.26ml dichloromethane, the PLA dichloromethane for forming a concentration of 320mg/ml of PLA is molten
Liquid；
(3) above two solution is mixed, is ultrasonically formed colostrum (o/w)；
(4) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution of 2L 0.5%, stirring forms multiple
Newborn (w/o/w)；
(5) emulsion (w/o/w) stirs, and removes dichloromethane, forms microballoon；
(6) it filters, removes PVA solution, form microballoon wet product；
(7) suitable mannitol solution is added, freeze-drying both obtains leuprorelin acetate microballoon.
Embodiment 11
(1) 225mg leuprorelin acetates are dissolved in 150ul waters for injection, it is 1.5g/ml's to form leuprolide concentrations
Leuprorelin aqueous solution for injection；
(2) 2g PLA are dissolved in 6.67ml dichloromethane, the PLA dichloromethane for forming a concentration of 300mg/ml of PLA is molten
Liquid；
(3) above two solution is mixed, is ultrasonically formed colostrum (o/w)；
(4) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution of 2L 0.5%, stirring forms multiple
Newborn (w/o/w)；
(5) emulsion (w/o/w) stirs, and removes dichloromethane, forms microballoon；
(6) it filters, removes PVA solution, form microballoon wet product；
(7) suitable mannitol solution is added, freeze-drying both obtains leuprorelin acetate microballoon.
Embodiment 12
(1) 225mg leuprorelin acetates are dissolved in 150ul waters for injection, it is 1.5g/ml's to form leuprolide concentrations
Leuprorelin aqueous solution for injection；
(2) 2g PLA are dissolved in 5.89ml dichloromethane, the PLA dichloromethane for forming a concentration of 340mg/ml of PLA is molten
Liquid；
(3) above two solution is mixed, is ultrasonically formed colostrum (o/w)；
(4) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution of 2L 0.5%, stirring forms multiple
Newborn (w/o/w)；
(5) emulsion (w/o/w) stirs, and removes dichloromethane, forms microballoon；
(6) it filters, removes PVA solution, form microballoon wet product；
(7) suitable mannitol solution is added, freeze-drying both obtains leuprorelin acetate microballoon.
Embodiment 13
(1) 225mg leuprorelin acetates are dissolved in water for injection, form bright third that leuprolide concentrations are 2.5g/ml
Rayleigh aqueous solution for injection；
(2) 2g PLA are dissolved in 5.71ml dichloromethane, the PLA dichloromethane for forming a concentration of 350mg/ml of PLA is molten
Liquid；
(3) above two solution is mixed, is ultrasonically formed colostrum (o/w)；
(4) colostrum (o/w) is added in PVA (Polyvinyl alcohol) solution of 2L 0.5%, stirring forms multiple
Newborn (w/o/w)；
(5) emulsion (w/o w) stirs, and removes dichloromethane, forms microballoon；
(6) it filters, removes PVA solution, form microballoon wet product.
Claims (8)
1. a kind of preparation method of sustained release leuprorelin acetate microballoon, it is characterised in that the step of preparation method is：
(1) leuprorelin acetate is dissolved in water for injection, forms leuprorelin acetate aqueous solution for injection；
(2) polylactic acid is dissolved in dichloromethane, forms polylactic acid dichloromethane solution；
(3) by the polylactic acid dichloromethane in the leuprorelin acetate aqueous solution for injection and the step (2) in the step (1)
Alkane solution mixes, and is ultrasonically formed colostrum；
(4) colostrum described in step (3) is added in poly-vinyl alcohol solution, stirring forms emulsion；
(5) emulsion is stirred, removes dichloromethane, form microballoon；
(6) it filters, removes poly-vinyl alcohol solution, form microballoon wet product；With
(7) mannitol solution is added into microballoon wet product, freeze-drying is to get sustained release leuprorelin acetate microballoon；
A concentration of 1g/ml-2.5g/ of leuprorelin acetate of leuprorelin acetate aqueous solution for injection described in the step (1)
ml；PLA concentration is 280mg/ml-350mg/ml in polylactic acid dichloromethane solution described in the step (2).
2. the preparation method of sustained release leuprorelin acetate microballoon as described in claim 1, it is characterised in that in the step (1)
A concentration of 1.5g/ml of leuprorelin acetate of the leuprorelin acetate aqueous solution for injection.
3. the preparation method of sustained release leuprorelin acetate microballoon as described in claim 1, it is characterised in that in the step (2)
PLA concentration is 320mg/ml in the polylactic acid dichloromethane solution.
4. the preparation method of sustained release leuprorelin acetate microballoon as described in claim 1, it is characterised in that in the step (2)
The weight average molecular weight of the polylactic acid is 12000-18000.
5. the preparation method of sustained release leuprorelin acetate microballoon as described in claim 1, it is characterised in that the acetic acid is bright
Third Rayleigh microballoon in vivo can sustained release 8 weeks or more.
6. the preparation method of sustained release leuprorelin acetate microballoon as claimed in claim 5, it is characterised in that the acetic acid is bright
Third Rayleigh microballoon in vivo can sustained release 12 weeks or so.
7. the preparation method of sustained release leuprorelin acetate microballoon as described in claim 1, it is characterised in that：
A concentration of 1.5g/ml of leuprorelin acetate of leuprorelin acetate aqueous solution for injection described in the step (1)；It is described
PLA concentration is 320mg/ml in polylactic acid dichloromethane solution described in step (2).
8. the preparation method of sustained release leuprorelin acetate microballoon as claimed in claim 1 or 7, it is characterised in that the step
(2) weight average molecular weight of polylactic acid described in is 12000-18000.
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| CN110420314A (en) * | 2018-04-27 | 2019-11-08 | 丽珠医药集团股份有限公司 | A kind of leuprorelin acetate slow releasing composition and preparation method thereof |
| CN109806385B (en) * | 2019-01-31 | 2022-04-08 | 浙江圣兆药物科技股份有限公司 | Leuprorelin acetate microsphere preparation and preparation method thereof |
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| CN113440597A (en) * | 2021-07-02 | 2021-09-28 | 山东谷雨春生物科技有限公司 | Method for preparing leuprorelin microspheres |
| CN114288384A (en) * | 2021-12-20 | 2022-04-08 | 海南海灵化学制药有限公司 | Preparation process of leuprorelin acetate sustained-release microspheres for injection |
| CN116270492A (en) * | 2023-03-30 | 2023-06-23 | 北京博恩特药业有限公司 | Leuprolide acetate sustained-release microsphere for injection and preparation method and application thereof |
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Inventor after: Lu Wenqi Inventor after: Wang Yanqing Inventor after: Kong Xiangsheng Inventor after: Xu Peng Inventor after: Liu Zhihui Inventor after: Cai Shimin Inventor after: Gao Tianyu Inventor after: Chen Bin Inventor before: Lu Wenqi Inventor before: Wang Yanqing Inventor before: Kong Xiangsheng Inventor before: Xu Peng Inventor before: Liu Zhihui Inventor before: Cai Shimin Inventor before: Gao Tianyu Inventor before: Chen Bin |