CN101879143B - Microsphere combination medicament containing antiparkinsonism drug and application thereof - Google Patents
Microsphere combination medicament containing antiparkinsonism drug and application thereof Download PDFInfo
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- CN101879143B CN101879143B CN 201010230676 CN201010230676A CN101879143B CN 101879143 B CN101879143 B CN 101879143B CN 201010230676 CN201010230676 CN 201010230676 CN 201010230676 A CN201010230676 A CN 201010230676A CN 101879143 B CN101879143 B CN 101879143B
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Abstract
The invention relates to an application of a microsphere composition containing an antiparkinsonism drug to preparation of a medicine for preventing and treating parkinson diseases and complicating diseases of parkinson diseases. The microsphere composition containing the antiparkinsonism drug is selected from benserazide microspheres, L-dopa methyl ester microspheres, the L-dopa methyl ester microspheres and the benserazide microspheres, or L-dopa methyl ester and benserazide mixing drug microspheres. The invention also provides a microsphere combination medicament containing the antiparkinsonism drug. The invention explores novel medical application for the microsphere combination medicament containing the antiparkinsonism drug and develops a novel application field. The microsphere combination medicament containing the antiparkinsonism drug of the invention is safe and nontoxic, has strong pharmacological action and indicates good medicinal prospect.
Description
[technical field]
The present invention relates to a kind of microsphere composition, specifically, is microsphere combination medicament and the application thereof that contains antiparkinsonism drug about a kind of.
[background technology]
At present to parkinson disease, symptomatic parkinsonism treatment mainly contains oral levodopa class medicine, daily requirement is taken three times, but for this class patient, be very inconvenient, because they took action originally and memory has problem, if can reach the effect in a week even month with a medicine, this is extraordinary something for them.
Chinese patent application 200910201414.X discloses a kind of selegiline sustained release microspheres and preparation method thereof.Chinese patent application numbers 200910201416.9 discloses a kind of rivastigmine slow-release microspheres and preparation method thereof.Chinese patent application numbers 200410030559.5 discloses levodopa nanometer formulation and preparation method thereof.Selegiline sustained release microspheres, rivastigmine slow-release microspheres and levodopa nanometer formulation are difficult to see through blood brain barrier and easily by intravital enzymatic degradation, intravital blood drug level is also lower than oral.Levodopa nanometer formulation instability, nanoparticle gathers easily, influences its drug effect, and the employed adjuvant of levodopa nanometer formulation is not pharmaceutic adjuvant, has toxic and side effects.
[summary of the invention]
The objective of the invention is at deficiency of the prior art, a kind of purposes that contains the microsphere composition of antiparkinsonism drug is provided.
One purpose more of the present invention is that a kind of microsphere combination medicament that contains antiparkinsonism drug is provided.
For realizing above-mentioned first purpose, the technical scheme that the present invention takes is:
A kind of application of microsphere composition in preparation prevention, treatment parkinson disease and parkinson disease complication disease medicament that contains antiparkinsonism drug, the described microsphere composition that contains antiparkinsonism drug is selected from benserazide microsphere, LDME microsphere, LDME microsphere and benserazide microsphere or LDME and benserazide mixed medicament microsphere.
Described microsphere composition said composition by weight percentage is composed of the following components:
Degradable hydrophobic polymer 50%-99%
Antiparkinsonism drug 1%-50%,
Described antiparkinsonism drug is selected from LDME or one or both mixture of benserazide.
Described degradable hydrophobic polymer is selected from a kind of or its mixture of polylactic acid-glycolic guanidine-acetic acid, polylactic acid or polycaprolactone.
Described microsphere composition particle diameter is 1-500 μ m.
Described microsphere composition particle diameter is 250-500 μ m.
Described microsphere composition is to obtain by oil-in-water-Water-In-Oil method (w/o/w), Water-In-Oil (o/w) method, the oil-in-water-solid method (s/o/w) of oil bag, the solid method (s/o/o) of oil bag oil-oil bag or spray drying method for preparation.
Described microsphere composition is to prepare by oil-in-water-Water-In-Oil method (w/o/w) or oil-in-water-solid method (s/o/w) of oil bag.
For realizing above-mentioned second purpose, the technical scheme that the present invention takes is:
A kind of microsphere combination medicament that contains antiparkinsonism drug, described composition of medicine is made up of LDME microsphere and benserazide microsphere.
Described LDME microsphere this microsphere by weight percentage is composed of the following components:
Degradable hydrophobic polymer 50%-99%
LDME 1%-50%,
Described benserazide microsphere this microsphere by weight percentage is composed of the following components:
Degradable hydrophobic polymer 50%-99%
Benserazide 1%-50%.
The invention has the advantages that:
1, the present invention utilizes oil-in-water method (W/O), oil-in-water-Water-In-Oil method (W/O/W), oil-in-water-solid method (S/O/W) of oil bag or the solid method (S/O/O) of oil bag oil-oil bag successfully to develop a kind of microsphere combination medicament that contains antiparkinsonism drug;
2, containing the microsphere combination medicament of antiparkinsonism drug can subcutaneous injection, intramuscular injection, lumbar injection or intracranial injection have long-term efficacy;
3, the microsphere combination medicament that contains antiparkinsonism drug has obvious synergism, and single with can significantly reducing the AIM scoring, two medicines can significantly reduce medical expense when being combined in the raising curative effect.
4, the present invention has excavated new medical application to the microsphere combination medicament that contains antiparkinsonism drug, has opened up a new application.
5, the microsphere combination medicament safety non-toxic that contains antiparkinsonism drug of the present invention, pharmacological action is strong, is indicating well prospect in medicine.
[description of drawings]
Fig. 1 LDME and benserazide mixed medicament slow-release PLGA microsphere composition sem photograph.
Fig. 2 LDME and benserazide mixed medicament slow-release PLGA microsphere composition release in vitro curve chart.
Fig. 3 L-dopa methyl ester sustained-release PLA microsphere composition sem photograph.
Fig. 4 L-dopa methyl ester sustained-release PLA microsphere composition release in vitro curve chart.
The benserazide sustained-release PLA microsphere composition sem photograph that Fig. 5 prepares with oil-in-water-Water-In-Oil method.
The benserazide sustained-release PLA microsphere composition release in vitro curve chart that Fig. 6 prepares with oil-in-water-Water-In-Oil method.
Fig. 7 wraps the benserazide sustained-release PLA microsphere composition sem photograph of solid method preparation with oil-in-water-oil.
Fig. 8 wraps the benserazide sustained-release PLA microsphere composition release in vitro curve chart of solid method preparation with oil-in-water-oil.
[specific embodiment]
Below in conjunction with accompanying drawing the specific embodiment provided by the invention is elaborated.
Embodiment 1
1. LDME and benserazide mixed medicament formulations prepared from solutions
A) 100mg LDME and the benserazide mixed medicament (is 1: 1 according to weight ratio) that market is bought, being mixed with weight percent concentration is 2.5%;
2. LDME and benserazide mixed medicament slow-release microsphere composition preparation
A) will be 1. LDME and benserazide mixed medicament solution measure 0.2mL, 0.5mL or 1mL respectively and take by weighing respectively 595mg polyglycolic acid-polylactic acid (the PLGA molecular weight is 6000), (the PLGA molecular weight is 250 to polyglycolic acid-polylactic acid of 37.5mg, 000) or polyglycolic acid-polylactic acid of 25mg (the PLGA molecular weight is 500,000) and be mixed with the organic solution that weight percent concentration is 30%, 15% or 5% dichloromethane respectively; Will be with 30% concentration corresponding one by one mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute with 0.2mL, 15% and order 0.5mL or 5% and above-mentioned solution 1mL; With the percentage composition that is prepared into LDME and benserazide mixed medicament is 1%, 25% or 50% sustained-release micro-spheres.
B) step (a) emulsion is added to the Polyethylene Glycol that weight percent concentration is 1% sodium chloride and 1% one by one respectively according to above-mentioned order (molecular weight of PVA is 1,000,000) (molecular weight of PVA is 500 for aqueous solution 10mL, 5% sodium chloride and 2.5% Polyethylene Glycol, 000) (molecular weight of PVA is 1 to the Polyethylene Glycol of aqueous solution 10mL or 10% sodium chloride and 2.5%, 000,000) formed emulsion in aqueous solution 10mL and stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 1%, 5% or 10% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, and (sem photograph of the microsphere of above-mentioned preparation as shown in Figure 1, smooth surface, the particle size distribution of microsphere are even, and particle diameter is about 40-100 μ m to obtain microsphere composition after the lyophilizing; Other particle diameter is about 60-150 μ m and 200-500 μ m respectively, does not all show on the figure).
The content that is prepared into LDME and benserazide mixed medicament be 1% microsphere at the release in vitro curve as shown in Figure 2, the result shows and not prominently releases and not exclusively discharge that other also has similar result but shows on the figure.
Stability test is investigated: the LDME and the benserazide mixed medicament slow-release microsphere of LDME and benserazide and the present invention's preparation are placed on radiation of visible light simultaneously, reclaim the content that detects LDME and benserazide respectively then, find LDME active decline 6-10% after a year, and the microsphere 0-1.0% that alters an agreement hardly; Benserazide is active decline 5-10% after 1 year, the 0-0.8% and microsphere is altered an agreement hardly.
Blood drug level is investigated in the body: be used for LDME and benserazide oral formulations and LDME and benserazide mixed medicament slow-release microsphere comparison with dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 30% in the benserazide body of LDME and benserazide mixed medicament slow-release microsphere dosage form; LDME and benserazide mixed medicament slow-release microsphere are found the LDME of microsphere dosage form also significantly better than oral formulations, and the gross area of blood drug level is higher than oral about 40% in its body.
Embodiment 2
1. LDME and benserazide mixed medicament formulations prepared from solutions
A) 100mg LDME and the benserazide mixed medicament (is 2: 1 according to weight ratio) that market is bought, being mixed with weight percent concentration is 2.5%;
2. LDME and benserazide mixed medicament slow-release microsphere composition preparation
A) will be 1. LDME and benserazide mixed medicament solution measure 0.2mL, 0.5mL or 1mL respectively and take by weighing respectively 595mg polylactic acid (the PLA molecular weight is 6000), (the PLA molecular weight is 250 to the polylactic acid of 37.5mg, 000) or the polylactic acid of 25mg (the PLA molecular weight is 500,000) and be mixed with the organic solution that weight percent concentration is 30%, 15% or 5% dichloromethane respectively; Will be with 30% concentration corresponding one by one mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute with 0.2mL, 15% and order 0.5mL or 5% and above-mentioned solution 1mL; With the percentage composition that is prepared into LDME and benserazide mixed medicament is 1%, 25% or 50% sustained-release micro-spheres.
B) step (a) emulsion is added to the Polyethylene Glycol that weight percent concentration is 1% sodium chloride and 1% respectively (molecular weight of PVA is 1,000,000) (molecular weight of PVA is 500 for aqueous solution 10mL, 5% sodium chloride and 2.5% Polyethylene Glycol, 000) (molecular weight of PVA is 1 to the Polyethylene Glycol of aqueous solution 10mL or 10% sodium chloride and 2.5%, 000,000) formed emulsion in aqueous solution 10mL and stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 1%, 5% or 10% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and wash with water 3-5 time, (sem photograph of the microsphere of above-mentioned preparation is as shown in Figure 1 similar, and smooth surface, the particle size distribution of microsphere are even, and particle diameter is about 45-100 μ m to obtain microsphere composition after the lyophilizing; Other particle diameter is about 60-170 μ m and 250-500 μ m respectively, does not all show on the figure).
To be 1% microsphere in the release in vitro result show the content that is prepared into LDME and benserazide mixed medicament not prominently releases and not exclusively discharge, and other also has similar result but shows on the figure.
Stability test is investigated: the LDME and the benserazide mixed medicament slow-release microsphere of LDME and benserazide and the present invention's preparation are placed on radiation of visible light simultaneously, reclaim the content that detects LDME and benserazide respectively then, find LDME active decline 6-8% after a year, and the microsphere 0-1.0% that alters an agreement hardly; Benserazide is active decline 5-9.1% after 1 year, the 0-0.78% and microsphere is altered an agreement hardly.
Blood drug level is investigated in the body: be used for LDME and benserazide oral formulations and LDME and benserazide mixed medicament slow-release microsphere comparison with dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 31% in the benserazide body of LDME and benserazide mixed medicament slow-release microsphere dosage form; LDME and benserazide mixed medicament slow-release microsphere are found the LDME of microsphere dosage form also significantly better than oral formulations, and the gross area of blood drug level is higher than oral about 42% in its body.
Embodiment 3
1. LDME and benserazide mixed medicament formulations prepared from solutions
A) 100mg LDME and the benserazide mixed medicament (is 3: 1 according to weight ratio) that market is bought, being mixed with weight percent concentration is 2.5%;
2. LDME and benserazide mixed medicament slow-release microsphere composition preparation
A) will be 1. LDME and benserazide mixed medicament solution measure 0.2mL, 0.5mL or 1mL respectively and take by weighing the polycaprolactone of 595mg respectively that (the PCL molecular weight is 10,000), (the PCL molecular weight is 2 to the polycaprolactone of 37.5mg, 500,000) or the polycaprolactone of 25mg (the PCL molecular weight is 5,000,000) and be mixed with the organic solution that weight percent concentration is 30%, 15% or 5% dichloromethane respectively; Will be with 30% concentration corresponding one by one mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute with 0.2mL, 15% and order 0.5mL or 5% and above-mentioned solution 1mL; With the percentage composition that is prepared into LDME and benserazide mixed medicament is 1%, 25% or 50% sustained-release micro-spheres.
B) step (a) emulsion is added to the Polyethylene Glycol that weight percent concentration is 1% sodium chloride and 1% respectively (molecular weight of PVA is 1,000,000) (molecular weight of PVA is 500 for aqueous solution 10mL, 5% sodium chloride and 2.5% Polyethylene Glycol, 000) (molecular weight of PVA is 1 to the Polyethylene Glycol of aqueous solution 10mL or 10% sodium chloride and 2.5%, 000,000) formed emulsion in aqueous solution 10mL and stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 1%, 5% or 10% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and wash with water 3-5 time, (sem photograph of the microsphere of above-mentioned preparation is as shown in Figure 1 similar, and smooth surface, the particle size distribution of microsphere are even, and particle diameter is about 50-100 μ m to obtain microsphere composition after the lyophilizing; Other particle diameter is about 65-150 μ m and 250-500 μ m respectively, does not all show on the figure).
To be 1% microsphere in the release in vitro result show the content that is prepared into LDME and benserazide mixed medicament not prominently releases and not exclusively discharge, and other also has similar result but shows on the figure.
Stability test is investigated: the LDME and the benserazide mixed medicament slow-release microsphere of LDME and benserazide and the present invention's preparation are placed on radiation of visible light simultaneously, reclaim the content that detects LDME and benserazide respectively then, find LDME active decline 6-7% after a year, and the microsphere 0-1.0% that alters an agreement hardly; Benserazide is active decline 5-9.5% after 1 year, the 0-0.72% and microsphere is altered an agreement hardly.
Blood drug level is investigated in the body: be used for LDME and benserazide oral formulations and LDME and benserazide mixed medicament slow-release microsphere comparison with dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 31.5% in the benserazide body of LDME and benserazide mixed medicament slow-release microsphere dosage form; LDME and benserazide mixed medicament slow-release microsphere are found the LDME of microsphere dosage form also significantly better than oral formulations, and the gross area of blood drug level is higher than oral about 42.5% in its body.
Embodiment 4
1. LDME and benserazide mixed medicament formulations prepared from solutions
A) 100mg LDME and the benserazide mixed medicament (is 4: 1 according to weight ratio) that market is bought, being mixed with weight percent concentration is 2.5%;
2. LDME and benserazide mixed medicament slow-release microsphere composition preparation
A) LDME that will 1. get and benserazide mixed medicament solution are measured 0.2mL, 0.5mL or 1mL respectively and are taken by weighing 100mg polyglycolic acid-polylactic acid (the PLGA molecular weight is 6000), 200mg polylactic acid (the PLA molecular weight is 6000) and 295mg polycaprolactone (the PCL molecular weight is 10, the 000) polymeric blends of 595mg altogether respectively; 12.5mg polyglycolic acid-polylactic acid (the PLGA molecular weight is 20,000), 15mg polylactic acid (the PLA molecular weight is 50,000) and 10mg polycaprolactone (the PCL molecular weight is 10,000) be the polymeric blends of 37.5mg altogether; Or 5mg polyglycolic acid-polylactic acid (the PLGA molecular weight is 50,000), 7mg polylactic acid (the PLA molecular weight is 500,000) and 13mg polycaprolactone (the PCL molecular weight is 1,000,000) are total to the polymeric blends of 25mg; And be mixed with the organic solution that weight percent concentration is 30%, 15% or 5% dichloromethane respectively; Will be with 30% concentration corresponding one by one mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute with 0.2mL, 15% and order 0.5mL or 5% and above-mentioned solution 1mL; It with the percentage composition that is prepared into LDME and benserazide mixed medicament 1%, 25% or 50% sustained-release micro-spheres.
B) step (a) emulsion is added to the Polyethylene Glycol that weight percent concentration is 1% sodium chloride and 1% respectively (molecular weight of PVA is 1,000,000) (molecular weight of PVA is 500 for aqueous solution 10mL, 5% sodium chloride and 2.5% Polyethylene Glycol, 000) (molecular weight of PVA is 1 to the Polyethylene Glycol of aqueous solution 10mL or 10% sodium chloride and 2.5%, 000,000) formed emulsion in aqueous solution 10mL and stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 1%, 5% or 10% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and wash with water 3-5 time, (sem photograph of the microsphere of above-mentioned preparation is as shown in Figure 1 similar, and smooth surface, the particle size distribution of microsphere are even, and particle diameter is about 55-100 μ m to obtain microsphere composition after the lyophilizing; Other particle diameter is about 65-150 μ m and 250-500 μ m respectively, does not all show on the figure).
To be 1% microsphere in the release in vitro result show the content that is prepared into LDME and benserazide mixed medicament not prominently releases and not exclusively discharge, and other also has similar result but shows on the figure.
Stability test is investigated: the LDME and the benserazide mixed medicament slow-release microsphere of LDME and benserazide and the present invention's preparation are placed on radiation of visible light simultaneously, reclaim the content that detects LDME and benserazide respectively then, find LDME active decline 6-7% after a year, and the microsphere 0-1.0% that alters an agreement hardly; Benserazide is active decline 5-8.5% after 1 year, the 0-0.72% and microsphere is altered an agreement hardly.
Blood drug level is investigated in the body: be used for LDME and benserazide oral formulations and LDME and benserazide mixed medicament slow-release microsphere comparison with dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 31.7% in the benserazide body of LDME and benserazide mixed medicament slow-release microsphere dosage form; LDME and benserazide mixed medicament slow-release microsphere are found the LDME of microsphere dosage form also significantly better than oral formulations, and the gross area of blood drug level is higher than oral about 43.5% in its body.
Need to prove embodiment 1,2,3,4th, LDME and the preparation of benserazide mixed medicament microsphere composition.
1. LDME formulations prepared from solutions
A), be mixed with weight percent concentration and be 2.5% aqueous solution with the 100mg LDME;
2. L-dopa methyl ester sustained-release microsphere composite preparation
(a) will take by weighing the polylactic acid (PLA of 37.5mg, molecular weight is 90,000-140,000) being mixed with weight percent concentration is the organic solution of 15% dichloromethane and to measure the above-mentioned 1. LDME solution of 0.5mL mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute; With the theoretical percentage composition that is prepared into LDME is 25% sustained-release micro-spheres.
(b) step (a) being got emulsion is added to the Polyethylene Glycol that weight percent concentration is 5% sodium chloride and 1% (molecular weight of PVA is 146,000-186,000, alcoholysis degree 98-99%) aqueous solution 10mL respectively and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 5% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, obtain after the lyophilizing microsphere composition (smooth surface, the particle size distribution of the microsphere of above-mentioned preparation are even, particle diameter for about 66-110 μ m as described in Figure 3).
The microsphere composition of preparation: the percentage by weight of its actual PLA is 75% and LDME is 25%, shake percentage ratio that external the 1st day burst size accounts for total LDME in the phosphate buffer solution of 37 ℃ and pH2 after being 20.63%, 14 day cumulative release 99.04%, and release profiles as described in Figure 4.
The envelop rate of the microsphere composition of method preparation of the present invention is equipped with the high 5-22% of microsphere than with the W/O legal system; First day prominent releasing than W/O and S/O/O method lacked 4%-11%.
Stability test is investigated: the LDME microsphere of LDME and the present invention's preparation is placed on radiation of visible light simultaneously, reclaim then and detect its content, find LDME active decline 6-10% after a year, and the microsphere 0-1.0% that alters an agreement hardly.
Therapeutic effect is investigated: be used for the LDME oral formulations of dosage and LDME microsphere relatively finding the microsphere dosage form significantly better than oral formulations that the gross area of blood drug level is higher than oral about 35% in its body.
Embodiment 6
1. LDME formulations prepared from solutions
A), be mixed with weight percent concentration and be 2.5% aqueous solution with the 100mg LDME;
2. L-dopa methyl ester sustained-release microsphere composite preparation
(a) will take by weighing the polylactic acid (PLA of 295mg, molecular weight is 90,000-140,000) being mixed with weight percent concentration is the organic solution of 15% dichloromethane and to measure the above-mentioned 1. LDME solution of 0.2mL mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute; With the theoretical percentage composition that is prepared into LDME is 1% sustained-release micro-spheres.
(b) step (a) being got emulsion is added to the Polyethylene Glycol that weight percent concentration is 5% sodium chloride and 1% (molecular weight of PVA is 146,000-186,000, alcoholysis degree 98-99%) aqueous solution 10mL respectively and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 10% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, obtains microsphere composition (smooth surface, the particle size distribution of the microsphere of above-mentioned preparation are even, and particle diameter is about 60-140 μ m) after the lyophilizing.
The microsphere composition of preparation: the percentage by weight of its actual PLA is 99% and LDME is 1%, shake percentage ratio that external the 1st day burst size accounts for total LDME in the phosphate buffer solution of 37 ℃ and pH3 after being 19.98%, 14 day cumulative release 92.23%.
The envelop rate of the microsphere composition of method preparation of the present invention is equipped with the high 5-19% of microsphere than with the W/O legal system; First day prominent releasing than W/O and S/O/O method lacked 4%-20%.
Stability test is investigated: the LDME microsphere of LDME and the present invention's preparation is placed on radiation of visible light simultaneously, reclaim then and detect its content, find LDME active decline 5-8% after a year, and the microsphere 0-0.8% that alters an agreement hardly.
Therapeutic effect is investigated: be used for the LDME oral formulations of dosage and LDME microsphere relatively finding the microsphere dosage form significantly better than oral formulations that the gross area of blood drug level is higher than oral about 36% in its body.
Embodiment 7
1. LDME formulations prepared from solutions
A) with the 100mg LDME, being mixed with weight percent concentration is 2.5%;
2. L-dopa methyl ester sustained-release microsphere composite preparation
A) will be 1. LDME solution measure 0.2mL, 0.5mL or 1mL respectively and take by weighing respectively 595mg polylactic acid (the PLA molecular weight is 6000), (the PLGA molecular weight is 250 to the polylactic acid of 37.5mg, 000) or the polylactic acid of 25mg (the PLGA molecular weight is 500,000) and be mixed with the organic solution that weight percent concentration is 30%, 15% or 5% dichloromethane respectively; Will be with 30% concentration corresponding one by one mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute with 0.2mL, 15% and order 0.5mL or 5% and above-mentioned LDME solution 1mL; With the percentage composition that is prepared into LDME is 1% or 50% sustained-release micro-spheres.
B) step (a) emulsion is added to the Polyethylene Glycol that weight percent concentration is 1% sodium chloride and 1% respectively (molecular weight of PVA is 1 according to above-mentioned order is corresponding one by one, 000,000) (molecular weight of PVA is 500 for aqueous solution 10mL, 5% sodium chloride and 2.5% Polyethylene Glycol, 000) (molecular weight of PVA is 1 to the Polyethylene Glycol of aqueous solution 10mL or 10% sodium chloride and 2.5%, 000,000) formed emulsion in aqueous solution 10mL and stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 5% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and wash with water 3-5 time, (sem photograph of the microsphere of above-mentioned preparation is as shown in Figure 3 similar, and smooth surface, the particle size distribution of microsphere are even, and particle diameter is about 45-100 μ m to obtain microsphere composition after the lyophilizing; Other particle diameter is about 60-170 μ m and 250-500 μ m respectively, does not all show on the figure).
The envelop rate of the microsphere composition of method preparation of the present invention is equipped with the high 4-12% of microsphere than with the W/O legal system; First day prominent releasing than W/O and S/O/O method lacked 2%-12%.
Stability test is investigated: the LDME microsphere of LDME and the present invention's preparation is placed on radiation of visible light simultaneously, reclaim then and detect its content, find LDME active decline 5-10% after a year, and the microsphere 0-0.8% that alters an agreement hardly.
Therapeutic effect is investigated: be used for comparing with the LDME oral formulations and the LDME microsphere of dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 20-36% in its body.
Embodiment 8
1. LDME formulations prepared from solutions
A) the 100mg LDME that market is bought, being mixed with weight percent concentration is 2.5%;
2. L-dopa methyl ester sustained-release microsphere composite preparation
A) will be 1. LDME solution measure 0.2mL, 0.5mL or 1mL respectively and take by weighing the polycaprolactone of 595mg respectively that (the PCL molecular weight is 10,000), (the PCL molecular weight is 2 to the polycaprolactone of 37.5mg, 500,000) or the polycaprolactone of 25mg (the PCL molecular weight is 5,000,000) and be mixed with the organic solution that weight percent concentration is 30%, 15% or 5% dichloromethane respectively; Will be with 30% concentration corresponding one by one mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute with 0.2mL, 15% and order 0.5mL or 5% and above-mentioned LDME solution 1mL; With the percentage composition that is prepared into LDME is 1%, 25% or 50% sustained-release micro-spheres.
B) step (a) emulsion is added to the Polyethylene Glycol that weight percent concentration is 1% sodium chloride and 1% one by one respectively according to above-mentioned order (molecular weight of PVA is 1,000,000) (molecular weight of PVA is 500 for aqueous solution 10mL, 5% sodium chloride and 2.5% Polyethylene Glycol, 000) (molecular weight of PVA is 1 to the Polyethylene Glycol of aqueous solution 10mL or 10% sodium chloride and 2.5%, 000,000) formed emulsion in aqueous solution 10mL and stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 1%, 5% or 10% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and wash with water 3-5 time, (sem photograph of the microsphere of above-mentioned preparation is as shown in Figure 3 similar, and smooth surface, the particle size distribution of microsphere are even, and particle diameter is about 50-100 μ m to obtain microsphere composition after the lyophilizing; Other particle diameter is about 65-150 μ m and 250-500 μ m respectively, does not all show on the figure).
The envelop rate of the microsphere composition of method preparation of the present invention is equipped with the high 4-13% of microsphere than with the W/O legal system; First day prominent releasing than W/O and S/O/O method lacked 2%-12%.
Stability test is investigated: the LDME microsphere of LDME and the present invention's preparation is placed on radiation of visible light simultaneously, reclaim then and detect its content, find LDME active decline 5-10% after a year, and the microsphere 0.1-1.0% that alters an agreement hardly.
Therapeutic effect is investigated: be used for comparing with the LDME oral formulations and the LDME microsphere of dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 20-36% in its body.
Embodiment 9
1. LDME formulations prepared from solutions
A) the 100mg LDME that market is bought, being mixed with weight percent concentration is 2.5%;
2. L-dopa methyl ester sustained-release microsphere composite preparation
A) the LDME solution that will 1. get is measured 0.2mL, 0.5mL or 1mL respectively and is taken by weighing 100mg polyglycolic acid-polylactic acid (the PLGA molecular weight is 6000), 200mg polylactic acid (the PLA molecular weight is 6000) and 295mg polycaprolactone (the PCL molecular weight is 10, the 000) polymeric blends of 595mg altogether respectively; 12.5mg polyglycolic acid-polylactic acid (the PLGA molecular weight is 20,000), 15mg polylactic acid (the PLA molecular weight is 50,000) and 10mg polycaprolactone (the PCL molecular weight is 10,000) be the polymeric blends of 37.5mg altogether; Or 5mg polyglycolic acid-polylactic acid (the PLGA molecular weight is 50,000), 7mg polylactic acid (the PLA molecular weight is 500,000) and 13mg polycaprolactone (the PCL molecular weight is 1,000,000) are total to the polymeric blends of 25mg; And be mixed with the organic solution that weight percent concentration is 30%, 15% or 5% dichloromethane respectively; Will be with 30% concentration corresponding one by one mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute with 0.2mL, 15% and order 0.5mL or 5% and above-mentioned LDME solution 1mL; It with the percentage composition that is prepared into LDME 1%, 25% or 50% sustained-release micro-spheres.
B) step (a) emulsion is added to the Polyethylene Glycol that weight percent concentration is 1% sodium chloride and 1% one by one respectively according to above-mentioned order (molecular weight of PVA is 1,000,000) (molecular weight of PVA is 500 for aqueous solution 10mL, 5% sodium chloride and 2.5% Polyethylene Glycol, 000) (molecular weight of PVA is 1 to the Polyethylene Glycol of aqueous solution 10mL or 10% sodium chloride and 2.5%, 000,000) formed emulsion in aqueous solution 10mL and stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 1%, 5% or 10% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and wash with water 3-5 time, (sem photograph of the microsphere of above-mentioned preparation is as shown in Figure 3 similar, and smooth surface, the particle size distribution of microsphere are even, and particle diameter is about 55-100 μ m to obtain microsphere composition after the lyophilizing; Other particle diameter is about 65-150 μ m and 250-500 μ m respectively, does not all show on the figure).
The envelop rate of the microsphere composition of method preparation of the present invention is equipped with the high 4-15% of microsphere than with the W/O legal system; First day prominent releasing than W/O and S/O/O method lacked 2%-12%.
Stability test is investigated: the LDME microsphere of LDME and the present invention's preparation is placed on radiation of visible light simultaneously, reclaim then and detect its content, find LDME active decline 5-10% after a year, and the microsphere 0.1-1.0% that alters an agreement hardly.
Therapeutic effect is investigated: be used for comparing with the LDME oral formulations and the LDME microsphere of dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 26-37% in its body.
The present invention utilizes the further microcapsule of oil-in-water-Water-In-Oil (W/O/W) preparation micro-sphere method to wrap in the macromolecular material with slow release.Make the microsphere features smooth surface rounding of its preparation, good evenness, solution regular without adhesion; The envelop rate height, prominent release little, the drug loading height.
Need to prove embodiment 5,6,7,8,9th, the preparation of LDME microsphere composition.
1. benserazide formulations prepared from solutions
A), be mixed with weight percent concentration and be 2.5% aqueous solution with the 100mg benserazide;
2. benserazide sustained-release microspherical composition preparation
(a) will take by weighing the polylactic acid (PLA of 37.5mg, molecular weight is 90,000-140,000) being mixed with weight percent concentration is the organic solution of 15% dichloromethane and to measure the above-mentioned 1. benserazide solution of 0.5mL mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute; With the theoretical percentage composition that is prepared into benserazide is 15% sustained-release micro-spheres.
(b) step (a) being got emulsion is added to the Polyethylene Glycol that weight percent concentration is 5% sodium chloride and 1% (molecular weight of PVA is 146,000-186,000, alcoholysis degree 98-99%) aqueous solution 10mL respectively and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 5% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, obtain after the lyophilizing microsphere composition (smooth surface, the particle size distribution of the microsphere of above-mentioned preparation are even, particle diameter for about 66-110 μ m as described in Figure 5).
The microsphere composition of preparation: the percentage by weight of its actual PLA is 85% and benserazide is 15%, shake percentage ratio that external the 1st hour burst size accounts for total benserazide in the phosphate buffer solution of 37 ℃ and pH2 after being 20.63%, 4 day cumulative release 99.04%, and release profiles as described in Figure 6.
The envelop rate of the microsphere composition of method preparation of the present invention is equipped with the high 5-30% of microsphere than with the W/O legal system; First day prominent releasing than W/O and S/O/O method lacked 3%-12%.
Stability test is investigated: the benserazide microsphere of benserazide and the present invention's preparation is placed on radiation of visible light simultaneously, reclaims then and detect its content, find benserazide active decline 5-10% after a year, and the microsphere 0-0.8% that alters an agreement hardly.
Blood drug level effect expedition in the body: be used for the benserazide oral formulations of dosage and benserazide microsphere relatively finding the microsphere dosage form that the gross area of blood drug level is higher than oral about 30% in its body significantly better than oral formulations.
1. benserazide formulations prepared from solutions
A), be mixed with weight percent concentration and be 2.5% aqueous solution with the 100mg benserazide;
2. benserazide sustained-release microspherical composition preparation
(a) will take by weighing the polylactic acid (PLA of 295mg, molecular weight is 90,000-140,000) being mixed with weight percent concentration is the organic solution of 15% dichloromethane and to measure the above-mentioned 1. benserazide solution of 0.2mL mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute; With the theoretical percentage composition that is prepared into benserazide is 1% sustained-release micro-spheres.
(b) step (a) being got emulsion is added to the Polyethylene Glycol that weight percent concentration is 5% sodium chloride and 1% (molecular weight of PVA is 146,000-186,000, alcoholysis degree 98-99%) aqueous solution 10mL respectively and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 10% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, obtains microsphere composition (smooth surface, the particle size distribution of the microsphere of above-mentioned preparation are even, and particle diameter is about 60-140 μ m) after the lyophilizing.
The microsphere composition of preparation: the percentage by weight of its actual PLA is 99% and benserazide is 1%, shake percentage ratio that external the 1st hour burst size accounts for total benserazide in the phosphate buffer solution of 37 ℃ and pH2 after being 18.88%, 7 day cumulative release 91.23%.
The envelop rate of the microsphere composition of method preparation of the present invention is equipped with the high 5-18% of microsphere than with the W/O legal system; First day prominent releasing than W/O and S/O/O method lacked 4%-18%.
Stability test is investigated: the benserazide microsphere of benserazide and the present invention's preparation is placed on radiation of visible light simultaneously, reclaims then and detect its content, find benserazide active decline 5-8% after a year, and the microsphere 0-0.8% that alters an agreement hardly.
Blood drug level is investigated in the body: be used for the benserazide oral formulations of dosage and benserazide microsphere relatively finding the microsphere dosage form significantly better than oral formulations that the gross area of blood drug level is higher than oral about 30% in its body.
Embodiment 12
1. benserazide formulations prepared from solutions
A), be mixed with weight percent concentration and be 2.5% aqueous solution with the 100mg benserazide;
2. benserazide sustained-release microspherical composition preparation
(a) will take by weighing the polylactic acid-glycolic guanidine-acetic acid (PLGA of 37.5mg, molecular weight is 6000-500,000) being mixed with weight percent concentration is the organic solution of 15% dichloromethane and to measure the above-mentioned 1. benserazide solution of 0.5mL mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute; With the theoretical percentage composition that is prepared into benserazide is 35% sustained-release micro-spheres.
(b) step (a) being got emulsion is added to the Polyethylene Glycol that weight percent concentration is 5% sodium chloride and 1% (molecular weight of PVA is 146,000-186,000, alcoholysis degree 98-99%) aqueous solution 10mL respectively and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 5% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, obtains microsphere composition (smooth surface, the particle size distribution of the microsphere of above-mentioned preparation are even, and particle diameter is about 66-110 μ m) after the lyophilizing.
The microsphere composition of preparation: the percentage by weight of its actual PLGA is 65% and benserazide is 35%, shake percentage ratio that external the 1st hour burst size accounts for total benserazide in the phosphate buffer solution of 37 ℃ and pH2 after being 20.63%, 4 day cumulative release 99.04%.
The envelop rate of the microsphere composition of method preparation of the present invention is equipped with the high 5-30% of microsphere than with the W/O legal system; First day prominent releasing than W/O and S/O/O method lacked 3%-12%.
Stability test is investigated: the benserazide microsphere of benserazide and the present invention's preparation is placed on radiation of visible light simultaneously, reclaims then and detect its content, find benserazide active decline 5-10% after a year, and the microsphere 0-0.8% that alters an agreement hardly.
Blood drug level is investigated in the body: be used for the benserazide oral formulations of dosage and benserazide microsphere relatively finding the microsphere dosage form significantly better than oral formulations that the gross area of blood drug level is higher than oral about 30% in its body.
Embodiment 13
1. benserazide formulations prepared from solutions
A), be mixed with weight percent concentration and be 2.5% aqueous solution with the 1000mg benserazide;
2. benserazide sustained-release microspherical composition preparation
(a) will take by weighing the polylactic acid (PLA of 25mg, molecular weight is 90,000-140,000) being mixed with weight percent concentration is the organic solution of 15% dichloromethane and to measure the above-mentioned 1. benserazide solution of 0.2mL mixed and formed even suspension, i.e. Water-In-Oil (W/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute; With the theoretical percentage composition that is prepared into benserazide is 45% sustained-release micro-spheres.
(b) step (a) being got emulsion is added to the Polyethylene Glycol that weight percent concentration is 5% sodium chloride and 1% (molecular weight of PVA is 146,000-186,000, alcoholysis degree 98-99%) aqueous solution 10mL respectively and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 10% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, obtains microsphere composition (smooth surface, the particle size distribution of the microsphere of above-mentioned preparation are even, and particle diameter is about 60-140 μ m) after the lyophilizing.
The microsphere composition of preparation: the percentage by weight of its actual PLA is 55% and benserazide is 45%, shake percentage ratio that external the 1st hour burst size accounts for total benserazide in the phosphate buffer solution of 37 ℃ and pH2 after being 18.88%, 7 day cumulative release 91.23%.
The envelop rate of the microsphere composition of method preparation of the present invention is equipped with the high 5-18% of microsphere than with the W/O legal system; First day prominent releasing than W/O and S/O/O method lacked 4%-18%.
Stability test is investigated: the benserazide microsphere of benserazide and the present invention's preparation is placed on radiation of visible light simultaneously, reclaims then and detect its content, find benserazide active decline 5-8% after a year, and the microsphere 0-0.8% that alters an agreement hardly.
Blood drug level is investigated in the body: be used for the benserazide oral formulations of dosage and benserazide microsphere relatively finding the microsphere dosage form significantly better than oral formulations that the gross area of blood drug level is higher than oral about 30% in its body.
Need to prove embodiment 10,11,12,13rd, the benserazide microsphere composition adopts oil-in-water-Water-In-Oil method preparation.
Embodiment 14
1. with the 100mg benserazide, earlier with microscopic examination whether at 0.4-10 μ m, if can not be ground into 0.4-5 μ m with pulverizer;
2. benserazide sustained-release microspherical composition preparation
(a) will take by weighing the polylactic acid (PLA of 37.5mg, molecular weight is 90,000-140,000) be mixed with weight percent concentration be the organic solution of 15% dichloromethane and take by weighing 1. benserazide microgranule 12.5mg mixed and formed even suspension, i.e. oil bag solid (S/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute; With the theoretical percentage composition that is prepared into benserazide is 25% sustained-release micro-spheres.
(b) step (a) being got emulsion is added to the Polyethylene Glycol that weight percent concentration is 5% sodium chloride and 1% (molecular weight of PVA is 146,000-186,000, alcoholysis degree 98-99%) aqueous solution 10mL respectively and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 5% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, obtain after the lyophilizing microsphere composition (smooth surface, the particle size distribution of the microsphere of above-mentioned preparation are even, particle diameter for about 66-120 μ m as described in Figure 7).
The microsphere composition of preparation: the percentage by weight of its actual PLA is 75% and benserazide is 25%, shake percentage ratio that external the 1st hour burst size accounts for total benserazide in the phosphate buffer solution of 37 ℃ and pH2 after being 10.63%, 7 day cumulative release 98.34%, and release profiles as described in Figure 8.
The envelop rate ratio of the microsphere composition of method preparation of the present invention is respectively than be equipped with the high 5-30% of microsphere with W/O and W/O/W legal system; First day prominent released than W/O and W/O/W method and S/O/O method and lacks 5%-20%.
Stability test is investigated: the benserazide microsphere of benserazide and the present invention's preparation is placed on radiation of visible light simultaneously, reclaims then and detect its content, find benserazide active decline 5-10% after a year, and the microsphere 0-0.8% that alters an agreement hardly.
Blood drug level is investigated in the body: be used for comparing with the benserazide oral formulations and the benserazide microsphere of dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 28-48% in its body.
1. benserazide microgranule preparation
With the 100mg benserazide, earlier with microscopic examination whether at 0.4-10 μ m, if can not be ground into 0.4-5 μ m with pulverizer;
2. benserazide sustained-release microspherical composition preparation
(a) will take by weighing polyglycolic acid-polylactic acid (PLGA, d, l-lactide:46-52%Mole and the glycolide 48-54%Mole of 495mg; Molecular weight is 5000-6000) be mixed with weight percent concentration be the organic solution of 30% dichloromethane and take by weighing 1. benserazide microgranule 5mg mixed and formed even suspension, i.e. oil bag solid (S/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute; With the theoretical percentage composition that is prepared into benserazide is 1% sustained-release micro-spheres.
(b) step (a) being got emulsion is added to the Polyethylene Glycol that weight percent concentration is 1% sodium chloride and 2% (molecular weight of PVA is 146,000-186,000, alcoholysis degree 98-99%) aqueous solution 10mL respectively and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 15% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, obtains microsphere composition (smooth surface, the particle size distribution of the microsphere of above-mentioned preparation are even, and particle diameter is about 70-100 μ m) after the lyophilizing.
The microsphere composition of preparation: the percentage by weight of its actual PLGA is 99% and benserazide is 1%, shake percentage ratio that external the 1st hour burst size accounts for total benserazide in the phosphate buffer solution of 37 ℃ and pH2 after being 10.63%, 4 day cumulative release 95.23%.
The envelop rate ratio of the microsphere composition of method preparation of the present invention is respectively than be equipped with the high 5%-30% of microsphere with W/O and W/O/W legal system; First day prominent released than W/O and W/O/W method and S/O/O method and lacks 5%-20%.
Stability test is investigated: the benserazide microsphere of benserazide and the present invention's preparation is placed on radiation of visible light simultaneously, reclaims then and detect its content, find benserazide active decline 5-10% after a year, and the microsphere 0-1% that alters an agreement hardly.
Blood drug level is investigated in the body: be used for comparing with the benserazide oral formulations and the benserazide microsphere of dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 27-46% in its body.
Embodiment 16
1. benserazide microgranule preparation
With the 100mg benserazide, earlier with microscopic examination whether at 0.4-10 μ m, if can not be ground into 0.4-5 μ m with pulverizer;
2. benserazide sustained-release microspherical composition preparation
(a) will take by weighing polyglycolic acid-polylactic acid (PLGA, d, l-lactide:46-52%Mole and the glycolide 48-54%Mole of 37.5mg; Molecular weight is 5000-6000) be mixed with weight percent concentration be the organic solution of 15% dichloromethane and take by weighing 1. benserazide microgranule 12.5mg mixed and formed even suspension, i.e. oil bag solid (S/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute; With the theoretical percentage composition that is prepared into benserazide is 25% sustained-release micro-spheres.
(b) step (a) being got emulsion is added to the Polyethylene Glycol that weight percent concentration is 5% sodium chloride and 1% (molecular weight of PVA is 146,000-186,000, alcoholysis degree 98-99%) aqueous solution 10mL respectively and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that 10% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, obtains microsphere composition (smooth surface, the particle size distribution of the microsphere of above-mentioned preparation are even, and particle diameter is about 60-120 μ m) after the lyophilizing.
The microsphere composition of preparation: the percentage by weight of its actual PLGA is 75% and benserazide is 25%, shake percentage ratio that external the 1st hour burst size accounts for total benserazide in 37 ℃ and pH are 2 phosphate buffer solution after being 32.88%, 4 day cumulative release 95.25%.
The envelop rate ratio of the microsphere composition of method preparation of the present invention is respectively than be equipped with the high 5-30% of microsphere with W/O and W/O/W legal system; First day prominent released than W/O and W/O/W method and S/O/O method and lacks 5%-20%.
Stability test is investigated: the benserazide microsphere of benserazide and the present invention's preparation is placed on radiation of visible light simultaneously, reclaims then and detect its content, find benserazide active decline 5-10% after a year, and the microsphere 0-0.9% that alters an agreement hardly.
Blood drug level is investigated in the body: be used for comparing with the benserazide oral formulations and the benserazide microsphere of dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 27-49% in its body.
Embodiment 17
1. with the 100mg benserazide, earlier with microscopic examination whether at 0.4-10 μ m, if can not be ground into 0.4-5 μ m with pulverizer;
2. benserazide sustained-release microspherical composition preparation
(a) will take by weighing polyglycolic acid-polylactic acid (PLGA, d, l-lactide:46-52%Mole and the glycolide 48-54%Mole of 25mg; Molecular weight is 6000) be mixed with weight percent concentration be the organic solution of 10% dichloromethane and take by weighing 1. benserazide microgranule 25mg mixed and formed even suspension, i.e. oil bag solid (S/O) emulsion in stirring, whirlpool or ultrasonic 1-5 minute; With the theoretical percentage composition that is prepared into benserazide is 50% sustained-release micro-spheres.
(b) step (a) emulsion is added to the Polyethylene Glycol that weight percent concentration is 1% sodium chloride and 1% respectively (molecular weight of PVA is 110,000-124,000, alcoholysis degree is alcoholysis degree 98-99%) aqueous solution 10mL and stirring, whirlpool or ultrasonic 0.1-5 minute formation emulsion;
(c) emulsion of step (b) being added to concentration is that 5% 1000mL sodium chloride solution solidified 1-4 hour;
(d) microsphere that step (c) is obtained carries out centrifugal collection, and washes with water 3-5 time, obtains microsphere composition (smooth surface, the particle size distribution of the microsphere of above-mentioned preparation are even, and particle diameter is about 50-120 μ m) after the lyophilizing.
The microsphere composition of preparation: the percentage by weight of its actual PLGA is 50% and benserazide is 50%, shake percentage ratio that external the 1st hour burst size accounts for total benserazide in 37 ℃ and pH are 2 phosphate buffer solution after being 45.78%, 4 day cumulative release 95.83%.
The envelop rate ratio of the microsphere composition of method preparation of the present invention is respectively than be equipped with the high 5-30% of microsphere with W/O and W/O/W legal system; First day prominent released than W/O and W/O/W method and S/O/O method and lacks 5%-20%.
Stability test is investigated: the benserazide microsphere of benserazide and the present invention's preparation is placed on radiation of visible light simultaneously, reclaims then and detect its content, find benserazide active decline 5-10% after a year, and the microsphere 0-0.8% that alters an agreement hardly.
Blood drug level is investigated in the body: be used for comparing with the benserazide oral formulations and the benserazide microsphere of dosage, find the microsphere dosage form significantly better than oral formulations, the gross area of blood drug level is higher than oral about 28-48% in its body.
Need to prove embodiment 14,15,16,17th, the benserazide microsphere composition adopts oil-in-water-oil bag solid method preparation.
Embodiment 18: parallel laboratory test
In order respectively to organize the performance of medicine below the scientific evaluation, we detect blood drug level and envelop rate in following each stability of organizing, the body simultaneously with the method for parallel test.
It is as follows that each organizes medicine grouping situation:
The A group: L-dopa methyl ester sustained-release microsphere adopts the method for W/O/W of the present invention to prepare, and wherein LDME content is 5%, and high polymer adjuvant content is 95%.
The B group: benserazide sustained-release microspherical adopts W/O/W method of the present invention to prepare, and wherein benserazide content is 5%, and high polymer adjuvant content is 95%.
The C group: benserazide sustained-release microspherical adopts (S/O/W) of the present invention method to prepare, and wherein benserazide content is 5%, and high polymer adjuvant content is 95%.
The D group: LDME and benserazide mixed medicament slow-release microsphere (W/O/W) method prepares, and wherein benserazide content is 2.5%, and LDME content is 2.5%, and high polymer adjuvant content is 95%.
The E group: drying means prepares selegiline sustained release microspheres in the O1/O2 emulsion of selegiline sustained release microspheres employing Chinese patent application 200910201414.X patent documentation report, wherein the Rivastigmine content in the selegiline sustained release microspheres is 5%, and high polymer adjuvant content is 95%.
The F group: drying means prepares rivastigmine slow-release microspheres in the O1/O2 emulsion of rivastigmine slow-release microspheres employing Chinese patent application number 200910201416.9 patent documentations report, wherein the Rivastigmine content in the rivastigmine slow-release microspheres is 5%, and high polymer adjuvant content is 95%.
The H group: the levodopa nanometer formulation adopts Chinese patent application number 200410030559.5 patent documentation reported method to prepare the levodopa nanometer formulation.
It is as follows that each organizes the drug study result:
The experiment of embodiment 19 purposes
1. the making of side PD rat model model partially
After the anesthesia of SD rat (180-220g) 3% pentobarbital aldehyde, razor cuts off the hair of head, successively with 2% iodine tincture and 75% cotton ball soaked in alcohol sterilization skin of head.Make the long otch of 6cm along sagittal suture, carefully peel off fascia and muscle, push periosteum open, expose sutura, dioxygen washing with 3% is clean, inserts the ear rod then, and the ear rod with a side inserts external auditory meatus gently earlier, after running at the bottom of the auditory meatus,external,bony that the ear rod is fixing, then same the insertion together of another rod of picking up the ears decided, the screw of gnathostat in the adjustment is filled in the last front tooth of rat in the groove of tooth fixing head, and install both sides eye socket fixed bar, screw whole screws at last.With reference to the rat whole brain stereotaxic atlas of Paxinos and Watson, coordinate is 3.7mm behind (1) bregma, sagittal suture right side 1.7mm, 7.8mm under the cranial periosteum, front tooth line-2.4mm; (2) 4.4mm behind the bregma, sagittal suture right side 1.2mm, 7.8mm under the cranial periosteum, front tooth line-2.4mm.6-OHDA (the fresh preparation of physiological saline solution contains 0.2% ascorbic acid, and concentration is 4mg/ml) two target spots are damaged a side striatum and are set up the inclined to one side side damage of PD rat model.Every some injection of microsyringe 6-OHDA 4ul, injection speed 1ul/min, let the acupuncture needle remain at a certain point 3 minutes.Injection apomorphine 0.5mg/kg (with the fresh preparation of normal saline that contains 1% ascorbic acid) in postoperative three all rat abdominal cavities, induce it to strong sideway swivel, weekly, each 30 minutes, continuously, number of revolutions is also keeping the person of stablizing to be considered as modeling success rat more than 7 times/minute.
2. experiment grouping
48 molding success parkinson rats are divided into eight groups at random: 6 every group.
First group: the PD group (the normal saline treatment, once a day, continuous two weeks)
Second group: benserazide group (10mg/kg, s.c. once a day, continuous two weeks)
The 3rd group: LDME group (10mg/kg, s.c. once a day, continuous two weeks),
The 4th group: LDME and benserazide group (LDME/kg of 5mg, benserazide/kg of s.c. and 5mg, s.c. once a day, continuous two weeks)
The 5th group: benserazide microsphere group 180mg/kg, subcutaneous injection, intramuscular injection, lumbar injection or cranial cavity injection once wherein contain benserazide 90mg in the 180mg benserazide microsphere),
The 6th group: LDME microsphere group (180mg/kg, subcutaneous injection, intramuscular injection, lumbar injection or cranial cavity injection once wherein contain LDME 90mg in the 180mg LDME microsphere),
The 7th group: LDME microsphere and benserazide microsphere group (70mg benserazide microsphere/kg, subcutaneous injection, intramuscular injection, lumbar injection or cranial cavity injection are once, wherein contain benserazide 35mg in the 70mg benserazide microsphere, 70mg LDME microsphere/kg, subcutaneous injection, intramuscular injection, lumbar injection or cranial cavity injection are once, wherein contain LDME 35mg in the 70mg LDME microsphere)
The 8th group: LDME and benserazide mixed medicament microsphere group (140mg/kg, subcutaneous injection, intramuscular injection, lumbar injection or cranial cavity injection once wherein contain benserazide 35mg and LDME 35mg in the mixed medicine microspheres of 140mg).
3. unusual involuntary movement (AIM) scoring
Carried out the AIM scoring at the 1st, 2,4,6,8,10,12,14 days that treat, every group of rat gives to carry out an AIM scoring every 20 minutes behind the corresponding treatment, carries out altogether 2 hours, observes 1 minute at every turn.Be divided into into 4 parts and mark (upper limb AIM, the AIM of actinal surface portion, axle property AIM and motion AIM), every part has or not with the order of severity according to it again and divides 5 grades (0-4): 0: do not have; 1:AIM existed less than 50% of observing time; 2:AIM existed greater than 50% of observing time; 3: continue to exist, stimulate to make to stop; 4: continue to exist, stimulation can not make it to stop.Each is organized unusual involuntary movement (AIM) appraisal result and sees Table 1
Table 1: each organizes unusual involuntary movement (AIM) appraisal result
| Each group | Upper limb AIM | The AIM of actinal surface portion | Axle property AIM | Motion AIM | Total points |
| First group | 4 | 4 | 4 | 4 | 16 |
| Second group | 4 | 4 | 4 | 4 | 16 |
| The 3rd group | 3 | 3 | 3 | 3 | 12 |
| The 4th group | 3 | 2 | 3 | 2 | 10 |
| The 5th group | 4 | 4 | 4 | 4 | 16 |
| The 6th group | 3 | 2 | 2 | 2 | 9 |
| The 7th group | 2 | 1 | 2 | 1 | 6 |
| The 8th group | 1 | 1 | 1 | 2 | 5 |
4. forelimb functional examination
We carry out the rat forelimb performance functional examination.Method is as follows: the experimenter is the fixing latter half of and hind leg of rat body on the other hand, makes it liftoff, and another hands is fixed a side forelimb lands another forelimb, with the oblique side shifting rat of rat forehand direction (mobile 90cm in the 5s), writes down the side forelimb step number that lands when mobile.Alternately measure the number that strides of both sides upper limb.The forelimb functional examination the results are shown in Table 2
Table 2 forelimb functional examination result
5. result
As can be known from the results of Table 1: the microsphere subcutaneous injection reduces various types of AIM scorings.As can be known from the results of Table 2: the forelimb coordination ability of 6-OHDA damage back rat descends, and its coordination ability rises behind the microsphere subcutaneous injection, and this unusual fluctuation disease that shows that levodopa causes has been hindered the coordination function of rat.But the microsphere subcutaneous injection obviously alleviates the decline of the rat coordination ability.
LDME can be passed through blood brain barrier, but LDME, LDME is micro-nano, and L-dopa methyl ester sustained-release microsphere is easily by enzymatic degradation in the body and inactivation.Though benserazide can not pass through blood brain barrier, can prevent LDME, by intravital enzymatic degradation, benserazide and LDME play collaborative effect.As can be known from the results of Table 1: (the 7th group: LDME microsphere and benserazide microsphere group of two medicine, the 8th group: LDME and benserazide mixed medicament microsphere group) obvious synergism arranged, single with (the 5th group: benserazide microsphere group, the 6th group: LDME microsphere group) can significantly reduce the AIM scoring, two medicines are combined in when improving curative effect can significantly reduce medical expense.
The above only is a preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the inventive method; can also make some improvement and replenish, these improvement and replenish and also should be considered as protection scope of the present invention.
Claims (6)
1. the application of microsphere composition in preparation prevention, treatment parkinson disease and parkinson disease complication disease medicament that contains antiparkinsonism drug, it is characterized in that, the described microsphere composition that contains antiparkinsonism drug is selected from LDME microsphere, LDME microsphere and benserazide microsphere or LDME and benserazide mixed medicament microsphere
Described microsphere composition said composition by weight percentage is composed of the following components:
Degradable hydrophobic polymer 50%-99%
Antiparkinsonism drug 1%-50%,
Described antiparkinsonism drug is selected from LDME or one or both mixture of benserazide,
Described degradable hydrophobic polymer is selected from a kind of or its mixture of polylactic acid-glycolic guanidine-acetic acid, polylactic acid or polycaprolactone.
2. application according to claim 1 is characterized in that: described microsphere composition particle diameter is 1-500 μ m.
3. application according to claim 1 is characterized in that: described microsphere composition particle diameter is 250-500 μ m.
4. application according to claim 1 is characterized in that: described microsphere composition is to obtain by oil-in-water-Water-In-Oil method (w/o/w), Water-In-Oil (o/w) method, the oil-in-water-solid method (s/o/w) of oil bag, the solid method (s/o/o) of oil bag oil-oil bag or spray drying method for preparation.
5. application according to claim 4 is characterized in that: described microsphere composition is to prepare by oil-in-water-Water-In-Oil method (w/o/w) or oil-in-water-solid method (s/o/w) of oil bag.
6. a microsphere combination medicament that contains antiparkinsonism drug is characterized in that, described composition of medicine is made up of LDME microsphere and benserazide microsphere,
Described LDME microsphere this microsphere by weight percentage is composed of the following components:
Degradable hydrophobic polymer 50%-99%
LDME 1%-50%,
Described benserazide microsphere this microsphere by weight percentage is composed of the following components:
Degradable hydrophobic polymer 50%-99%
Benserazide 1%-50%
Described degradable hydrophobic polymer is selected from a kind of or its mixture of polylactic acid-glycolic guanidine-acetic acid, polylactic acid or polycaprolactone.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1682704A (en) * | 2004-04-14 | 2005-10-19 | 中南大学 | Levodopa nano preparation and its preparing method |
| CN101283965A (en) * | 2007-10-26 | 2008-10-15 | 南京长澳医药科技有限公司 | Preparation containing bendopa methyl ester and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1682704A (en) * | 2004-04-14 | 2005-10-19 | 中南大学 | Levodopa nano preparation and its preparing method |
| CN101283965A (en) * | 2007-10-26 | 2008-10-15 | 南京长澳医药科技有限公司 | Preparation containing bendopa methyl ester and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| Betul Arica等.Carbidopa/levodopa-loaded biodegradable microspheres:in vivo evaluation on experimental Parkinsonism in rats.《Journal of Controlled Release》.2004,第102卷第689-697页. * |
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