CA2247013A1 - Microencapsulation or drugs - Google Patents
Microencapsulation or drugs Download PDFInfo
- Publication number
- CA2247013A1 CA2247013A1 CA 2247013 CA2247013A CA2247013A1 CA 2247013 A1 CA2247013 A1 CA 2247013A1 CA 2247013 CA2247013 CA 2247013 CA 2247013 A CA2247013 A CA 2247013A CA 2247013 A1 CA2247013 A1 CA 2247013A1
- Authority
- CA
- Canada
- Prior art keywords
- drug
- polyhydroxyalkanoate
- aqueous medium
- coenzyme
- synthesized
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/62—Carboxylic acid esters
- C12P7/625—Polyesters of hydroxy carboxylic acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1682—Processes
- A61K9/1694—Processes resulting in granules or microspheres of the matrix type containing more than 5% of excipient
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5089—Processes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
- B01J13/02—Making microcapsules or microballoons
- B01J13/06—Making microcapsules or microballoons by phase separation
- B01J13/14—Polymerisation; cross-linking
- B01J13/18—In situ polymerisation with all reactants being present in the same phase
- B01J13/185—In situ polymerisation with all reactants being present in the same phase in an organic phase
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1641—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
- A61K9/1647—Polyesters, e.g. poly(lactide-co-glycolide)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5005—Wall or coating material
- A61K9/5021—Organic macromolecular compounds
- A61K9/5031—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poly(lactide-co-glycolide)
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Manufacturing Of Micro-Capsules (AREA)
- Medicinal Preparation (AREA)
Abstract
A drug, hydrophilic or lipophilic is entrapped or microencapsulated in a poly(hydroxyalkanoate) homopolymer or copolymer employing in vitro synthesis of the homopolymer or copolymer in an aqueous medium containing the drug which is to be entrapped or microencapsulated.
Description
BACKGROUND OF THE INVENTION
i) Field of the Invention This invention relates to a process for producing an entrapped or microencapsulated drug, more especially a process employing in vitro synthesis of a poiyhydroxyalkanoate homopolyrner or copolymer.
ii) Description of Prior Art Poly(hydroxyalkanoates), PHA, are biopoly((3-hydroxyalkanoate)-esters which are produced by microorganisms. The general pathways of PHA biosynthesis are well known. The poly((3-hydroxybutyrate) (PHB) biosynthetic pathway uses acetyl-CoA which generates [R]-(-)-3-hydr~xybutyryl-CoA as the natural monomer. This monomer responds to the naturally produced polymerise enzyme which rapidly produces PHB.
Processes have been proposed for encapsulating drugs within a polyhydroxyalkanoate shell. These processes t5rpically require use of organic solvents, see for example Bissery et al, Microspheres and Drug Therapy Pharmaceutical, Immunological and Medical Aspects, edited by S. S. Davis et al, Elsevier Science, pp. 217-227, 1984; Juni et al Journal of Controlled Release, 4, 2S-32, 1986; and Kawaguchi et al, Journal of Pharmaceutical Sciences, 81, S08-512, 1992. Use of polyhydroxyalkanoates in the entrappment or microencapsulation of drugs is especially advantageous since these polymers are biodegradable, non-toxic and otherwise non-harmful when administered to a living body.
It would be desirable to provide new entrappment or encapsulation processes employing non-toxic, biodegradable polymers, especially polymers that can be formed in situ in the entrappment or encapsulation of the drug.
SUMMARY OF THE INVENTION
This invention seeks to provide a process for producing an entrapped or microencapsulated drug.
In particular, this invention seeks to provide such a process for the entrappment of hydrophilic drugs.
In particular, this invention seeks to provide such a process for the microencapsulation of lipophilic drugs.
i) Field of the Invention This invention relates to a process for producing an entrapped or microencapsulated drug, more especially a process employing in vitro synthesis of a poiyhydroxyalkanoate homopolyrner or copolymer.
ii) Description of Prior Art Poly(hydroxyalkanoates), PHA, are biopoly((3-hydroxyalkanoate)-esters which are produced by microorganisms. The general pathways of PHA biosynthesis are well known. The poly((3-hydroxybutyrate) (PHB) biosynthetic pathway uses acetyl-CoA which generates [R]-(-)-3-hydr~xybutyryl-CoA as the natural monomer. This monomer responds to the naturally produced polymerise enzyme which rapidly produces PHB.
Processes have been proposed for encapsulating drugs within a polyhydroxyalkanoate shell. These processes t5rpically require use of organic solvents, see for example Bissery et al, Microspheres and Drug Therapy Pharmaceutical, Immunological and Medical Aspects, edited by S. S. Davis et al, Elsevier Science, pp. 217-227, 1984; Juni et al Journal of Controlled Release, 4, 2S-32, 1986; and Kawaguchi et al, Journal of Pharmaceutical Sciences, 81, S08-512, 1992. Use of polyhydroxyalkanoates in the entrappment or microencapsulation of drugs is especially advantageous since these polymers are biodegradable, non-toxic and otherwise non-harmful when administered to a living body.
It would be desirable to provide new entrappment or encapsulation processes employing non-toxic, biodegradable polymers, especially polymers that can be formed in situ in the entrappment or encapsulation of the drug.
SUMMARY OF THE INVENTION
This invention seeks to provide a process for producing an entrapped or microencapsulated drug.
In particular, this invention seeks to provide such a process for the entrappment of hydrophilic drugs.
In particular, this invention seeks to provide such a process for the microencapsulation of lipophilic drugs.
In accordance with the invention there is provided a process for producing an entrapped or microencapsulated drug comprising: providing a drug in an aqueous medium, synthesizing a polyhydroxyalkanoate homopolymer or copolymer in said aqueous medium by in vitro enzyme polymerization of at least one hydroxyalkanoate-Coenzyme A monomer precursor of said polyhydroxyalkanoate, with. formation in said aqueous medium of microporous granules or microcapsules of the synthesized polyhydroxyalkanoate entrapping or microencapsulating fluid from said medium, containing said drug, and recovering said granules or microcapsules from said aqueous medium.
DETAILED DESCRIPTION OF THE INVENTION
i) Polyhydroxyalkanoate Homop~lymer and t:opolymers The polyhydroxyalkanoate homoholymers and copolymers are derived from hydroxyalkanoates of 3 to 14 carbon atoms, The homopolymers are based on a single hydroxyalkanoate, and the copolymers are based on two different hydroxyalkanoates.
The preferred hydroxyalkanoates are the 3-hydroxybutyrates and the 3-hydroxyvalerates. The preferred homopolymers are derived from the 3-hydroxybutyrates, namely, poly((3-hydroxybutylate), i.e., PHB; the preferred copolymers are derived from 3-hydroxybut3zate and 3-hydroxyvalerate, namely, poly(3-hydroxybutyrate-co-3-hydroxyva.lerate) i.e., P(3HB-3HV).
The homopolymers and copolymers derived from hydroxyalkanoates in accordance with the invention are non-toxic and biodegradable. For example, poly(3-hydroxybutyrate) degrades to (D)-3-hydroxybutyric acid which is a normal constituent of human blood being produced during fatty acid oxidation in the liver. Complete degradation in the environment leads to Co2 and H20.
The PHB homopolymer is brittle so that in some situations there may be a preference for the P(3HB-3HV) copolymer which is more ductile.
Suitable copolymers may contain 10 to 15%, preferably 11 to 13%, by weight, of 3 hydroxyvalerate.
The preferred alkanoates are the R.-(-)-3-hydroxyalkanoic acids and the homopolymers and copolymers produc~;d by the in vitro synthesis of the invention having a weight average molecular weight of 10x106 to 1.3x10.
The homopolymers and copolymers produced by the in vitro synthesis are non-crystalline and the polymer granules produced are essentially spherical, typically having a size of 1 to 4, more especially 2 to 3 Vim.
ii) In vitro synthesis The biosynthesis of PHA is well established and PHA is produced in vivo by more than 90 genera of bacteria and different biosyntheses pathways are described by Jackson et al Annals of the New York Academy of Sciences, 745, (Biochem. Eng. VIII) 134-148, 1994. Representative bacteria for the in vivo biosynthesis are Alca,li eyes eutrc, R,~,odos irillum rubrurn, $seudomol~s oleovorans and ~'seudomonas a~inosa. In each case the polymerization is catalyzed by a PHA synthase, also referred to as PHA
polymerase and the substrate monomer is a llydroxyacyl-Coenzyme A, in which the acyl corresponds to the desired alkanoate.
Recently an in vitro synthesis has been employed which does not require the bacteria employed in the biosynthesis.
The in vitro synthesis employs the synthetically produced hydroxy acyl-Coenzyme A substrate monomer and a polyhydroxyalkanoate polymerase expressed from an isolated structured gene; for example, the polymerase may be encoded by a gene which is expressed naturally by ~Alcaligenes eutro us or Pseudolno~~as oleovoran_s.
The genes encoding the polyhydroxyalkanoate polymerase which occur naturally in PHA producing bacteria such .as l~c a es e~tr~hu~ have been identified and isolated and expressed in a non-PHA producing organism, . coli. The modified . oli does not produce PHA but does produce the polyhydroxyalkanoate polymerase. This polymerase is employed in the in vitro synthesis of PHA illustrated in Equation (I) below for the in vitro synthesis of poly(3-hydroxybutyrate):
CH3-(CH(OH)-CH2-CO-CoA~ PE~B polYmera CH3_(CH(OH)-CH2-CO [ O-CH(C:H3)-CH2-CO j-n O-CH(CH3)-CH2-CO-OH + CoA (I) in which n is an integer of the order of 200,000 to 150,000, CoA' refers to the mercapto radical formed by removing hydrogen from the thiol of the thioethylamino of Coenzyme A, and CoA refers ~:o Coenzyme A. Coenzyme A
is a well established co-factor in enzymatic acetyl transfer reactions and is described with formula in The Merck Index, Eleventh Edition, published by Merck & Co. Inc. 1989, entry No. 2465 at pages 385-386. Coenzyme A is a molecule which can be considered to be built u~? from p~ntetheine, adenosine and phosphoric acid, the empirical formula is C21H36N7016P3s and the chemical formula is:
~2 N ~ N
N
N
O C'H 2 O=P-OH ' ' H H
0 OH' O 0=P-OH
OH
I
O = P OCIiZ- C- CHCONHC1H2CHZCONHCHZCH2SH
DETAILED DESCRIPTION OF THE INVENTION
i) Polyhydroxyalkanoate Homop~lymer and t:opolymers The polyhydroxyalkanoate homoholymers and copolymers are derived from hydroxyalkanoates of 3 to 14 carbon atoms, The homopolymers are based on a single hydroxyalkanoate, and the copolymers are based on two different hydroxyalkanoates.
The preferred hydroxyalkanoates are the 3-hydroxybutyrates and the 3-hydroxyvalerates. The preferred homopolymers are derived from the 3-hydroxybutyrates, namely, poly((3-hydroxybutylate), i.e., PHB; the preferred copolymers are derived from 3-hydroxybut3zate and 3-hydroxyvalerate, namely, poly(3-hydroxybutyrate-co-3-hydroxyva.lerate) i.e., P(3HB-3HV).
The homopolymers and copolymers derived from hydroxyalkanoates in accordance with the invention are non-toxic and biodegradable. For example, poly(3-hydroxybutyrate) degrades to (D)-3-hydroxybutyric acid which is a normal constituent of human blood being produced during fatty acid oxidation in the liver. Complete degradation in the environment leads to Co2 and H20.
The PHB homopolymer is brittle so that in some situations there may be a preference for the P(3HB-3HV) copolymer which is more ductile.
Suitable copolymers may contain 10 to 15%, preferably 11 to 13%, by weight, of 3 hydroxyvalerate.
The preferred alkanoates are the R.-(-)-3-hydroxyalkanoic acids and the homopolymers and copolymers produc~;d by the in vitro synthesis of the invention having a weight average molecular weight of 10x106 to 1.3x10.
The homopolymers and copolymers produced by the in vitro synthesis are non-crystalline and the polymer granules produced are essentially spherical, typically having a size of 1 to 4, more especially 2 to 3 Vim.
ii) In vitro synthesis The biosynthesis of PHA is well established and PHA is produced in vivo by more than 90 genera of bacteria and different biosyntheses pathways are described by Jackson et al Annals of the New York Academy of Sciences, 745, (Biochem. Eng. VIII) 134-148, 1994. Representative bacteria for the in vivo biosynthesis are Alca,li eyes eutrc, R,~,odos irillum rubrurn, $seudomol~s oleovorans and ~'seudomonas a~inosa. In each case the polymerization is catalyzed by a PHA synthase, also referred to as PHA
polymerase and the substrate monomer is a llydroxyacyl-Coenzyme A, in which the acyl corresponds to the desired alkanoate.
Recently an in vitro synthesis has been employed which does not require the bacteria employed in the biosynthesis.
The in vitro synthesis employs the synthetically produced hydroxy acyl-Coenzyme A substrate monomer and a polyhydroxyalkanoate polymerase expressed from an isolated structured gene; for example, the polymerase may be encoded by a gene which is expressed naturally by ~Alcaligenes eutro us or Pseudolno~~as oleovoran_s.
The genes encoding the polyhydroxyalkanoate polymerase which occur naturally in PHA producing bacteria such .as l~c a es e~tr~hu~ have been identified and isolated and expressed in a non-PHA producing organism, . coli. The modified . oli does not produce PHA but does produce the polyhydroxyalkanoate polymerase. This polymerase is employed in the in vitro synthesis of PHA illustrated in Equation (I) below for the in vitro synthesis of poly(3-hydroxybutyrate):
CH3-(CH(OH)-CH2-CO-CoA~ PE~B polYmera CH3_(CH(OH)-CH2-CO [ O-CH(C:H3)-CH2-CO j-n O-CH(CH3)-CH2-CO-OH + CoA (I) in which n is an integer of the order of 200,000 to 150,000, CoA' refers to the mercapto radical formed by removing hydrogen from the thiol of the thioethylamino of Coenzyme A, and CoA refers ~:o Coenzyme A. Coenzyme A
is a well established co-factor in enzymatic acetyl transfer reactions and is described with formula in The Merck Index, Eleventh Edition, published by Merck & Co. Inc. 1989, entry No. 2465 at pages 385-386. Coenzyme A is a molecule which can be considered to be built u~? from p~ntetheine, adenosine and phosphoric acid, the empirical formula is C21H36N7016P3s and the chemical formula is:
~2 N ~ N
N
N
O C'H 2 O=P-OH ' ' H H
0 OH' O 0=P-OH
OH
I
O = P OCIiZ- C- CHCONHC1H2CHZCONHCHZCH2SH
The production of polyhydroxyall~;anoate polymerase for use in the in vitro present invention is described in U.S. Patent 5,229,279, Anthony Sinskey et al, issued July 20, 1993 and U.S. Patent 5,250,430, Anthony Sinskey et al, issued October 5, 1993. Furthermore, polyhydroxybutyrate polymerase produced by the procedures descrit~ed in the two U.S. Patents is available from Metabolix Inc. of Cambridge, Massachusetts, U.S.A.
The monomer substrate employed in the in vitro process of the invention is produced by chemical reaction between Coenzyme A and the appropriate hydroxyalkanoic acid, the reaction. being an esterification type reaction between the carboxylic acid group of the; hydroxyalkanoic acid and the thiol group of the Coenzyme A.
The in vitro synthesis of the invention is carried out in an aqueous medium containing the drug to be microencapsulated by the in vitro synthesized hornopolymer or copolymer.
Typically a phosphate buffer and the synthetic 3-hydroxyalkanoate-CoA are introduced into the aqueous medium and the polymerization is initiated by the addition ~of the polyhydroxyalkanoate polymerase. The synthesis may be followed spectrophotometrically, following a 200-fold dilution, by monitoring the disappearance of the 3-hydroxyalkanoate-CoA and the release of free CoA, by following the absorptions at 260 and 236 nm. Typically the reaction is completed in about 2 hours. The free Coenzyme A is recovered and may be employed to synthesize fresh 3-hydroxyalkanoate-CoA monomer substrate for polymerization.
The formed granules or microcapsules are recovered from the aqueous medium and dried.
In the in vitro synthesis the aveoage granule or microcapsule diameter, particle size distribution and volume increase with reaction time;
the average granule or microcapsule size increases rapidly initially, especially during the first 20 minutes of reaction.
Since the in vitro synthesis is free of components present in the bacterial cell in the in vivo synthesis, for example, lipids and proteins, the granule or microcapsule size is significantly larger than achieved in in vivo synthesis and additionally coagulation occurs early in the reaction. In the in vivo synthesis bacterial cell companents such as lipids and proteins stabilize the granules against coagulation.
The monomer substrate employed in the in vitro process of the invention is produced by chemical reaction between Coenzyme A and the appropriate hydroxyalkanoic acid, the reaction. being an esterification type reaction between the carboxylic acid group of the; hydroxyalkanoic acid and the thiol group of the Coenzyme A.
The in vitro synthesis of the invention is carried out in an aqueous medium containing the drug to be microencapsulated by the in vitro synthesized hornopolymer or copolymer.
Typically a phosphate buffer and the synthetic 3-hydroxyalkanoate-CoA are introduced into the aqueous medium and the polymerization is initiated by the addition ~of the polyhydroxyalkanoate polymerase. The synthesis may be followed spectrophotometrically, following a 200-fold dilution, by monitoring the disappearance of the 3-hydroxyalkanoate-CoA and the release of free CoA, by following the absorptions at 260 and 236 nm. Typically the reaction is completed in about 2 hours. The free Coenzyme A is recovered and may be employed to synthesize fresh 3-hydroxyalkanoate-CoA monomer substrate for polymerization.
The formed granules or microcapsules are recovered from the aqueous medium and dried.
In the in vitro synthesis the aveoage granule or microcapsule diameter, particle size distribution and volume increase with reaction time;
the average granule or microcapsule size increases rapidly initially, especially during the first 20 minutes of reaction.
Since the in vitro synthesis is free of components present in the bacterial cell in the in vivo synthesis, for example, lipids and proteins, the granule or microcapsule size is significantly larger than achieved in in vivo synthesis and additionally coagulation occurs early in the reaction. In the in vivo synthesis bacterial cell companents such as lipids and proteins stabilize the granules against coagulation.
iii) Hydrophilic Drugs In the case of hydrophilic drugs., for example, netilmicin and dopamine (a-methyldopa), the drugs are dissolved in the aqueous medium, which then forms the aqueous medium for the t'rc vitro synthesis. In this case the in vitro synthesis produces polymer particles which coalesce to microporous granules which entraps the aqueous medium containing the mlcropores.
In particular, the synthesized polymer particles are allowed to coalesce to form the microporous granules, and the granules are recovered with aqueous medium and dissolved drug entrapped in the micropores of the microporous granules. The granules are dried with the entrapped drug therein.
iv) Lipophilic Drugs In the case of lipophilic drugs, fir example, progesterone, the drugs are dissolved in an oil, which is then emulsified or dispersed in an aqueous medium as an oil-in-water emulsion or dispersion. The aqueous medium forms an aqueous phase containing droplets of the oil and the drug is contained in the oil droplets. In this case the in vitro synthesized polymer forms a microcapsule wall or shell about a fluid core, the fluid core being of the oil containing the dissolved lipophilic drug. The forrrled microcapsules are recovered from the aqueous phase and dried.
BRIEF DESCRIPTION OF DRAWINGS
FIG. 1 illustrates schematically eni:rappment of a lipophilic drug by a process of the invention, and FIG. 2 illustrates schematically microencapsulation of a hydrophilic drug by a process of the invention.
DESCRIPTION OF PREFERRED EMBODIMENTS
WITH REFERENCE TO THE DRAWINGS
With further reference to Fig. 1, there is shown schematically a flow diagram for the microencapsulation of a lipophilic drug such as progesterone by the in vitro synthesis of poly(3-hydroxybutyrate) in accordance with the invention.
The lipophilic drug is dissolved in an oil phase. An aqueous phase is added to the oil phase and the two phases are sonicated to produce an oil-in-water emulsion in which the dispersed or emulsified oil droplets contain the drug. The 3-hydroxybutyrate-Coenzyme A and the polyhydroxybutyrate polymerase are added to the emulsion where they enter the aqueous phase and poly(3-hydroxybutyrate) is synthesized around the oil droplets. The encapsulated oil droplets are then removed from the emulsion and dried to provide microcapsules having a shell of the p~oly(3-hydroxybutyrate) and a fluid core comprising the oil with the lipophilic drug dissolved therein. It will be understood that the oil should be one suitable for oral administration to a human, for example, a vegetable oil such as olive oil.
With further reference to Fig. 2, there is shown a flow sheet for the production of polymer granules for entrappment of a hydrophilic drug such as dopamine.
The hydrophilic drug is dissolved in an aqueous medium to form an aqueous solution of the drug. The 3-hydroxybutyrate-Coenzyme A and the polyhydroxybutyrate polymerase are added to the: aqueous solution, the poly(3-hydroxybutyrate) is synthesized within the aqueous solution as granules which coalesce forming a microporous granule which entraps the aqueous solution in the micropares. In this way there are produced polymer granules entrapping a fluid comprising the drug in aqueous solution. The polymer granules are removed from the aqueous solution end dried.
The in vitro synthesis of the invention may be characterized as a precipitation polymerization or pseudoemulsion ~~olymerization.
EXAMPLLS
Example 1 Netilmicin, a water soluble antibiotic, was assayed according to Emit procedure. To three test tubes a constant volume of 3 aqueous solutions of netilmicin were added, namely, aqueous solutions of 34 micrograms per litre, 5.0 micrograms per litre and 3.0 micrograms per litre of netilmicin. To each test tube was added a constant amount of PHB polymerase and 3-hydroxybutyrate-Coenzyme A monomer substrate and the reaction was allowed to proceed overnight. After centrifugation to remove the PHB in vitro particles with their content of netilmicin, an assay of the netilmicin and the supernatant was made. From the results it was found that 17%, 4.1 % and 2.1 %
of the total initial netilmicin was entrapped in the in vitro particles.
-g_ Example 2 The Netilmicin Emit assay was used. To 3 Eppendorf centrifuge tubes each containing 1 mg of netilmicin dissolved in water were added 1 micro litre, 2 micro lures and 5 miicro litres of PHB polymerise solution and the same amount of substrate monomer solution (3-hydroxybutyrate-Coenzyme A). After standing overnight the tubes were centrifuged at 14,000 rpm for 10 minutes and the supernatant was assayed for neti.lmicin. Tn addition, the pellets in the centrifuge tubes were dissolved in 300 micro litres of chloroform and 500 micro litres of water. The pellets were found to captain 0.42%, 1.2% and 4.5% of the total initial nedlmicin.
Example 3 A radio-labelled lipophilic steroid drug (tritiated progesterone) was dissolved in olive oil and an excess of water was added to create an oil-in-water emulsion. PHB polymerise ind monomer substrate (3-hydroxybutyrate-Coenzyme A) were added to the emulsion with gentle stirring. After allowing the reaction to proceed overnight a suspension of particles was obtained. The particles were hardened by adding an aqueous polyvinyl alcohol solution with stirring, and the suspension was collected on a Millipore (Trade-mark) filter.
Analytical radioassay showed that steroid and PHN were predominantly present in the solid collected on the filter.
In particular, the synthesized polymer particles are allowed to coalesce to form the microporous granules, and the granules are recovered with aqueous medium and dissolved drug entrapped in the micropores of the microporous granules. The granules are dried with the entrapped drug therein.
iv) Lipophilic Drugs In the case of lipophilic drugs, fir example, progesterone, the drugs are dissolved in an oil, which is then emulsified or dispersed in an aqueous medium as an oil-in-water emulsion or dispersion. The aqueous medium forms an aqueous phase containing droplets of the oil and the drug is contained in the oil droplets. In this case the in vitro synthesized polymer forms a microcapsule wall or shell about a fluid core, the fluid core being of the oil containing the dissolved lipophilic drug. The forrrled microcapsules are recovered from the aqueous phase and dried.
BRIEF DESCRIPTION OF DRAWINGS
FIG. 1 illustrates schematically eni:rappment of a lipophilic drug by a process of the invention, and FIG. 2 illustrates schematically microencapsulation of a hydrophilic drug by a process of the invention.
DESCRIPTION OF PREFERRED EMBODIMENTS
WITH REFERENCE TO THE DRAWINGS
With further reference to Fig. 1, there is shown schematically a flow diagram for the microencapsulation of a lipophilic drug such as progesterone by the in vitro synthesis of poly(3-hydroxybutyrate) in accordance with the invention.
The lipophilic drug is dissolved in an oil phase. An aqueous phase is added to the oil phase and the two phases are sonicated to produce an oil-in-water emulsion in which the dispersed or emulsified oil droplets contain the drug. The 3-hydroxybutyrate-Coenzyme A and the polyhydroxybutyrate polymerase are added to the emulsion where they enter the aqueous phase and poly(3-hydroxybutyrate) is synthesized around the oil droplets. The encapsulated oil droplets are then removed from the emulsion and dried to provide microcapsules having a shell of the p~oly(3-hydroxybutyrate) and a fluid core comprising the oil with the lipophilic drug dissolved therein. It will be understood that the oil should be one suitable for oral administration to a human, for example, a vegetable oil such as olive oil.
With further reference to Fig. 2, there is shown a flow sheet for the production of polymer granules for entrappment of a hydrophilic drug such as dopamine.
The hydrophilic drug is dissolved in an aqueous medium to form an aqueous solution of the drug. The 3-hydroxybutyrate-Coenzyme A and the polyhydroxybutyrate polymerase are added to the: aqueous solution, the poly(3-hydroxybutyrate) is synthesized within the aqueous solution as granules which coalesce forming a microporous granule which entraps the aqueous solution in the micropares. In this way there are produced polymer granules entrapping a fluid comprising the drug in aqueous solution. The polymer granules are removed from the aqueous solution end dried.
The in vitro synthesis of the invention may be characterized as a precipitation polymerization or pseudoemulsion ~~olymerization.
EXAMPLLS
Example 1 Netilmicin, a water soluble antibiotic, was assayed according to Emit procedure. To three test tubes a constant volume of 3 aqueous solutions of netilmicin were added, namely, aqueous solutions of 34 micrograms per litre, 5.0 micrograms per litre and 3.0 micrograms per litre of netilmicin. To each test tube was added a constant amount of PHB polymerase and 3-hydroxybutyrate-Coenzyme A monomer substrate and the reaction was allowed to proceed overnight. After centrifugation to remove the PHB in vitro particles with their content of netilmicin, an assay of the netilmicin and the supernatant was made. From the results it was found that 17%, 4.1 % and 2.1 %
of the total initial netilmicin was entrapped in the in vitro particles.
-g_ Example 2 The Netilmicin Emit assay was used. To 3 Eppendorf centrifuge tubes each containing 1 mg of netilmicin dissolved in water were added 1 micro litre, 2 micro lures and 5 miicro litres of PHB polymerise solution and the same amount of substrate monomer solution (3-hydroxybutyrate-Coenzyme A). After standing overnight the tubes were centrifuged at 14,000 rpm for 10 minutes and the supernatant was assayed for neti.lmicin. Tn addition, the pellets in the centrifuge tubes were dissolved in 300 micro litres of chloroform and 500 micro litres of water. The pellets were found to captain 0.42%, 1.2% and 4.5% of the total initial nedlmicin.
Example 3 A radio-labelled lipophilic steroid drug (tritiated progesterone) was dissolved in olive oil and an excess of water was added to create an oil-in-water emulsion. PHB polymerise ind monomer substrate (3-hydroxybutyrate-Coenzyme A) were added to the emulsion with gentle stirring. After allowing the reaction to proceed overnight a suspension of particles was obtained. The particles were hardened by adding an aqueous polyvinyl alcohol solution with stirring, and the suspension was collected on a Millipore (Trade-mark) filter.
Analytical radioassay showed that steroid and PHN were predominantly present in the solid collected on the filter.
Claims (11)
1. A process for producing an entrapped or microencapsulated drug comprising:
providing a drug in an aqueous medium, synthesizing a polyhydroxyalkanoate homopolymer or copolymer in said aqueous medium by in vitro enzyme polymerization of at least one hydroxyalkanoate Coenzyme A monomer being a precursor of said polyhydroxyalkanoate, with formation in said aqueous medium of microporous granules or microcapsules having a capsule wall of the synthesized polyhydroxyalkanoate entrapping or microencapsulating a fluid from said medium, containing said drug, and recovering said granules or microcapsules from said aqueous medium.
providing a drug in an aqueous medium, synthesizing a polyhydroxyalkanoate homopolymer or copolymer in said aqueous medium by in vitro enzyme polymerization of at least one hydroxyalkanoate Coenzyme A monomer being a precursor of said polyhydroxyalkanoate, with formation in said aqueous medium of microporous granules or microcapsules having a capsule wall of the synthesized polyhydroxyalkanoate entrapping or microencapsulating a fluid from said medium, containing said drug, and recovering said granules or microcapsules from said aqueous medium.
2. A process according to claim 1, wherein said drug is hydrophilic and is dissolved in said aqueous medium, said fluid comprising said aqueous medium with said drug dissolved therein, said fluid being entrapped in microporous granules of the synthesized polyhydroxyalkanoate.
3. A process according to claim 1, wherein said drug is lipophilic, and droplets of oil containing said drug are dispersed in said aqueous medium, said fluid comprises said oil with said drug dissolved therein, said fluid forming a fluid core within microcapsules of the synthesized polyhydroxyalkanoate.
4. A process according to claim 1, 2 or 3, wherein said precursor monomer is selected from 3-hydroxybutyryl-Coenzyme A, 3-hydroxyvalerate-Coenzyme A and mixtures thereof, and said enzyme polymerization is catalyzed by a corresponding polyhydroxyalkanoate synthase.
5. A process according to claim 1, 2 or 3, wherein said precursor monomer is (R)-(-)-3-hydroxybutyryl-Coenzyme A and said synthesized polyhydroxyalkanoate is of polyhydroxybutyrate homopolymer.
6. A process according to claim 2, wherein said precursor monomer is (R)-(-)-3-hydroxybutyryl-Coenzyme A and said synthesized polyhydroxyalkanoate is of polyhydroxybutyrate homopolymer.
7. A process according to claim 3, wherein said precursor monomer is (R)-(-)-3-hydroxybutyryl-Coenzyme A and said synthesized polyhydroxyalkanoate is of polyhydroxybutyrate homopolymer.
8. A process for producing a drug entrapped particle comprising:
dissolving a hydrophilic drug in an aqueous medium, synthesising a polyhydroxyalkanoate homopolymer or copolymer in said aqueous medium by in vitro enzyme polymerization of at least one hydroxyalkanoate Coenzyme A monomer which is a precursor of said polyhydroxyalkanoate, with formation in said aqueous medium of particles of the synthesized polyhydroxyalkanoate, allowing said particles to coalesce to microporous granules, recovering said microporous granules with said aqueous medium and dissolved hydrophilic drug entrapped in the micropores of said microporous granules, and drying said granules with said entrapped hydrophilic drug.
dissolving a hydrophilic drug in an aqueous medium, synthesising a polyhydroxyalkanoate homopolymer or copolymer in said aqueous medium by in vitro enzyme polymerization of at least one hydroxyalkanoate Coenzyme A monomer which is a precursor of said polyhydroxyalkanoate, with formation in said aqueous medium of particles of the synthesized polyhydroxyalkanoate, allowing said particles to coalesce to microporous granules, recovering said microporous granules with said aqueous medium and dissolved hydrophilic drug entrapped in the micropores of said microporous granules, and drying said granules with said entrapped hydrophilic drug.
9. A process according to claim 8, wherein said precursor monomer is (R)-(-)-3-hydroxybutyryl-Coenzyme A and said synthesized polyhydroxyalkanoate is of polyhydroxybutyrate homopolymer.
10. A process for producing a microencapsulated lipophilic drug comprising:
dissolving a lipophilic drug in an edible oil, adding an aqueous medium and forming an oil-in-water emulsion comprising droplets of said oil containing the dissolved lipophilic drug, in an aqueous phase, synthesizing a polyhydroxyalkanoate homopolymer or copolymer in said aqueous phase by in vitro enzyme polymerization of at least one hydroxyalkanoate Coenzyme A monomer which is a precursor of said polyhydroxyalkanoate, with formation in said aqueous phase of microcapsules having a capsule wall of the synthesized polyhydroxyalkanoate entrapping a fluid core comprising said oil with said drug dissolved therein, recovering said microcapsules, and drying said microcapsules with said fluid core therein.
dissolving a lipophilic drug in an edible oil, adding an aqueous medium and forming an oil-in-water emulsion comprising droplets of said oil containing the dissolved lipophilic drug, in an aqueous phase, synthesizing a polyhydroxyalkanoate homopolymer or copolymer in said aqueous phase by in vitro enzyme polymerization of at least one hydroxyalkanoate Coenzyme A monomer which is a precursor of said polyhydroxyalkanoate, with formation in said aqueous phase of microcapsules having a capsule wall of the synthesized polyhydroxyalkanoate entrapping a fluid core comprising said oil with said drug dissolved therein, recovering said microcapsules, and drying said microcapsules with said fluid core therein.
11. A process according to claim 10, wherein said precursor monomer is (R)-(-)-3-hydroxybutyryl-Coenzyme A and said synthesized polyhydroxyalkanoate is of polyhydroxybutyrate homopolymer.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA 2247013 CA2247013A1 (en) | 1998-09-09 | 1998-09-09 | Microencapsulation or drugs |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA 2247013 CA2247013A1 (en) | 1998-09-09 | 1998-09-09 | Microencapsulation or drugs |
Publications (1)
Publication Number | Publication Date |
---|---|
CA2247013A1 true CA2247013A1 (en) | 2000-03-09 |
Family
ID=29409895
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA 2247013 Abandoned CA2247013A1 (en) | 1998-09-09 | 1998-09-09 | Microencapsulation or drugs |
Country Status (1)
Country | Link |
---|---|
CA (1) | CA2247013A1 (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9814657B2 (en) | 2009-04-27 | 2017-11-14 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
US10434047B2 (en) | 2009-04-27 | 2019-10-08 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
-
1998
- 1998-09-09 CA CA 2247013 patent/CA2247013A1/en not_active Abandoned
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9814657B2 (en) | 2009-04-27 | 2017-11-14 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
US10434044B2 (en) | 2009-04-27 | 2019-10-08 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
US10434046B2 (en) | 2009-04-27 | 2019-10-08 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
US10434047B2 (en) | 2009-04-27 | 2019-10-08 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
US10675227B2 (en) | 2009-04-27 | 2020-06-09 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
US10688026B2 (en) | 2009-04-27 | 2020-06-23 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
US11185479B2 (en) | 2009-04-27 | 2021-11-30 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
US11185480B2 (en) | 2009-04-27 | 2021-11-30 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
US11185478B2 (en) | 2009-04-27 | 2021-11-30 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
US11850294B2 (en) | 2009-04-27 | 2023-12-26 | Premier Dental Products Company | Buffered microencapsulated compositions and methods |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US6146665A (en) | Entrapment or microencapsulation of drugs in a polyhydroxyalkanoate formed by enzyme synthesis | |
JP5372310B2 (en) | Agglomeration of encapsulated microcapsules and production thereof | |
EP1275378B1 (en) | Particulate construct comprising polyhydroxyalkanoate and method for producing it | |
JP3990880B2 (en) | Method for producing polyhydroxyalkanoate-coated liposome | |
CN104004134B (en) | A kind of preparation method of the monodispersity arch nanospheres of size tunable | |
NO315692B1 (en) | New microcapsule, as well as injectable composition containing the microcapsule, process for the preparation of a contrast agent, and a single process for the preparation of said microcapsules | |
JP2004174492A (en) | Method of stabilizing enzyme and active ingredient using polyol/high molecular microcapsule and cosmetic composition containing the same | |
US5851452A (en) | Process for coating droplets or nanometric particles | |
US7527809B2 (en) | Polyhydroxyalkanoate-containing magnetic structure, and manufacturing method and use thereof | |
JP4510842B2 (en) | Polyhydroxyalkanoate-coated liposome | |
CA2247013A1 (en) | Microencapsulation or drugs | |
CN113426389B (en) | Preparation method of alcohol soluble protein microcapsule and product | |
JP2003175092A (en) | Granular body containing polyhydroxy-alkanoate and method of making the same and application of the same | |
Liu et al. | Current status and challenges in the application of microbial PHA particles | |
El-Gibaly et al. | Microencapsulation of ketoprofen using w/o/w complex emulsion technique | |
AU2006201070B2 (en) | Encapsulated agglomeration of microcapsules and method for the preparation thereof | |
CN117243925A (en) | PEG-PLGA nanoparticle coated with clotrimazole and application thereof | |
Cedrati et al. | Preparation and characterisation of poly (lactic acid) hemoglobin microspheres | |
JPS63170437A (en) | Production of microsponge | |
Tuladhar et al. | Recent Advances in Applications of Encapsulation Technology for the Bioprotection of Phytonutrients in Complex Food Systems | |
Rasiah et al. | Biopolyester particles: preparation and applications | |
CN116570591A (en) | Schisandrin B and regorafenib composition, preparation method and application thereof | |
JP2004512405A (en) | Process for the preparation of microsphere-like crystallites from linear polysaccharides, the corresponding microsphere-like crystallites and their use | |
Gallo et al. | Mincroencapsulation as a New Approach to Protect Active Compounds in Food |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request | ||
FZDE | Dead |