CA2238352C - Process for the production of morphologically uniform microcapsules and microcapsules that are produced according to this process - Google Patents
Process for the production of morphologically uniform microcapsules and microcapsules that are produced according to this process Download PDFInfo
- Publication number
- CA2238352C CA2238352C CA002238352A CA2238352A CA2238352C CA 2238352 C CA2238352 C CA 2238352C CA 002238352 A CA002238352 A CA 002238352A CA 2238352 A CA2238352 A CA 2238352A CA 2238352 C CA2238352 C CA 2238352C
- Authority
- CA
- Canada
- Prior art keywords
- solution
- microcapsules
- solvent
- process according
- polymer
- 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.)
- Expired - Lifetime
Links
- 238000000034 method Methods 0.000 title claims abstract description 62
- 239000003094 microcapsule Substances 0.000 title claims abstract description 45
- 238000004519 manufacturing process Methods 0.000 title claims description 7
- 239000000243 solution Substances 0.000 claims abstract description 57
- 239000002904 solvent Substances 0.000 claims abstract description 31
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 24
- 239000007864 aqueous solution Substances 0.000 claims abstract description 23
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 16
- 239000000839 emulsion Substances 0.000 claims abstract description 13
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 13
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 13
- 239000004094 surface-active agent Substances 0.000 claims abstract description 9
- 239000000203 mixture Substances 0.000 claims abstract description 8
- 239000013543 active substance Substances 0.000 claims abstract description 7
- 229920002988 biodegradable polymer Polymers 0.000 claims abstract description 5
- 239000004621 biodegradable polymer Substances 0.000 claims abstract description 5
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 34
- 229920000642 polymer Polymers 0.000 claims description 34
- 239000004480 active ingredient Substances 0.000 claims description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 17
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims description 16
- 229920001577 copolymer Polymers 0.000 claims description 12
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 10
- -1 C1-C4 alkyl acetates Chemical class 0.000 claims description 9
- 102000004877 Insulin Human genes 0.000 claims description 8
- 108090001061 Insulin Proteins 0.000 claims description 8
- 239000002202 Polyethylene glycol Substances 0.000 claims description 8
- 229940125396 insulin Drugs 0.000 claims description 8
- 229920001223 polyethylene glycol Polymers 0.000 claims description 8
- 239000011877 solvent mixture Substances 0.000 claims description 8
- 102000008100 Human Serum Albumin Human genes 0.000 claims description 7
- 108091006905 Human Serum Albumin Proteins 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 7
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical class OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 claims description 6
- 229920000728 polyester Polymers 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 5
- 229920001400 block copolymer Polymers 0.000 claims description 5
- JMMWKPVZQRWMSS-UHFFFAOYSA-N isopropanol acetate Natural products CC(C)OC(C)=O JMMWKPVZQRWMSS-UHFFFAOYSA-N 0.000 claims description 5
- 229940011051 isopropyl acetate Drugs 0.000 claims description 5
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 claims description 4
- 108010050904 Interferons Proteins 0.000 claims description 4
- 102000014150 Interferons Human genes 0.000 claims description 4
- 229940124041 Luteinizing hormone releasing hormone (LHRH) antagonist Drugs 0.000 claims description 4
- GWYFCOCPABKNJV-UHFFFAOYSA-N isovaleric acid Chemical compound CC(C)CC(O)=O GWYFCOCPABKNJV-UHFFFAOYSA-N 0.000 claims description 4
- URAYPUMNDPQOKB-UHFFFAOYSA-N triacetin Chemical compound CC(=O)OCC(OC(C)=O)COC(C)=O URAYPUMNDPQOKB-UHFFFAOYSA-N 0.000 claims description 4
- KXKVLQRXCPHEJC-UHFFFAOYSA-N acetic acid trimethyl ester Natural products COC(C)=O KXKVLQRXCPHEJC-UHFFFAOYSA-N 0.000 claims description 3
- 239000008055 phosphate buffer solution Substances 0.000 claims description 3
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 claims description 2
- DOOTYTYQINUNNV-UHFFFAOYSA-N Triethyl citrate Chemical compound CCOC(=O)CC(O)(C(=O)OCC)CC(=O)OCC DOOTYTYQINUNNV-UHFFFAOYSA-N 0.000 claims description 2
- 239000007979 citrate buffer Substances 0.000 claims description 2
- 239000001087 glyceryl triacetate Substances 0.000 claims description 2
- 235000013773 glyceryl triacetate Nutrition 0.000 claims description 2
- 229940047124 interferons Drugs 0.000 claims description 2
- 229960002622 triacetin Drugs 0.000 claims description 2
- 239000001069 triethyl citrate Substances 0.000 claims description 2
- VMYFZRTXGLUXMZ-UHFFFAOYSA-N triethyl citrate Natural products CCOC(=O)C(O)(C(=O)OCC)C(=O)OCC VMYFZRTXGLUXMZ-UHFFFAOYSA-N 0.000 claims description 2
- 235000013769 triethyl citrate Nutrition 0.000 claims description 2
- 108010005714 Interferon beta-1b Proteins 0.000 claims 1
- 229940021459 betaseron Drugs 0.000 claims 1
- 238000005538 encapsulation Methods 0.000 abstract description 14
- 239000008307 w/o/w-emulsion Substances 0.000 abstract description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 20
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 18
- 239000000725 suspension Substances 0.000 description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 16
- 239000004372 Polyvinyl alcohol Substances 0.000 description 14
- 229920002451 polyvinyl alcohol Polymers 0.000 description 14
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 10
- 229910052757 nitrogen Inorganic materials 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- 239000003795 chemical substances by application Substances 0.000 description 7
- 238000005354 coacervation Methods 0.000 description 7
- 239000006185 dispersion Substances 0.000 description 7
- 239000003960 organic solvent Substances 0.000 description 6
- 229910000831 Steel Inorganic materials 0.000 description 5
- 239000008346 aqueous phase Substances 0.000 description 5
- 238000001704 evaporation Methods 0.000 description 5
- 230000008020 evaporation Effects 0.000 description 5
- 229920000036 polyvinylpyrrolidone Chemical class 0.000 description 5
- 239000001267 polyvinylpyrrolidone Chemical class 0.000 description 5
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 5
- 239000010959 steel Substances 0.000 description 5
- 235000000346 sugar Nutrition 0.000 description 5
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 4
- 229920000954 Polyglycolide Polymers 0.000 description 4
- 229940088597 hormone Drugs 0.000 description 4
- 239000005556 hormone Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 229920001987 poloxamine Polymers 0.000 description 4
- 229920000747 poly(lactic acid) Polymers 0.000 description 4
- 229920000098 polyolefin Polymers 0.000 description 4
- 229920000136 polysorbate Polymers 0.000 description 4
- 229920002545 silicone oil Polymers 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 150000005846 sugar alcohols Chemical class 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- JJTUDXZGHPGLLC-UHFFFAOYSA-N lactide Chemical compound CC1OC(=O)C(C)OC1=O JJTUDXZGHPGLLC-UHFFFAOYSA-N 0.000 description 3
- 239000011159 matrix material Substances 0.000 description 3
- 238000005191 phase separation Methods 0.000 description 3
- 229920001432 poly(L-lactide) Polymers 0.000 description 3
- 229920001606 poly(lactic acid-co-glycolic acid) Polymers 0.000 description 3
- 238000001694 spray drying Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 229920003178 (lactide-co-glycolide) polymer Polymers 0.000 description 2
- RKDVKSZUMVYZHH-UHFFFAOYSA-N 1,4-dioxane-2,5-dione Chemical compound O=C1COC(=O)CO1 RKDVKSZUMVYZHH-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- 101000904173 Homo sapiens Progonadoliberin-1 Proteins 0.000 description 2
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 2
- 102100024028 Progonadoliberin-1 Human genes 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical class O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 101000996723 Sus scrofa Gonadotropin-releasing hormone receptor Proteins 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- NMJJFJNHVMGPGM-UHFFFAOYSA-N butyl formate Chemical compound CCCCOC=O NMJJFJNHVMGPGM-UHFFFAOYSA-N 0.000 description 2
- LZCLXQDLBQLTDK-UHFFFAOYSA-N ethyl 2-hydroxypropanoate Chemical compound CCOC(=O)C(C)O LZCLXQDLBQLTDK-UHFFFAOYSA-N 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- XLXSAKCOAKORKW-UHFFFAOYSA-N gonadorelin Chemical compound C1CCC(C(=O)NCC(N)=O)N1C(=O)C(CCCN=C(N)N)NC(=O)C(CC(C)C)NC(=O)CNC(=O)C(NC(=O)C(CO)NC(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C(CC=1NC=NC=1)NC(=O)C1NC(=O)CC1)CC1=CC=C(O)C=C1 XLXSAKCOAKORKW-UHFFFAOYSA-N 0.000 description 2
- 239000002474 gonadorelin antagonist Substances 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 229940079322 interferon Drugs 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 239000012074 organic phase Substances 0.000 description 2
- 229920000620 organic polymer Polymers 0.000 description 2
- 230000003285 pharmacodynamic effect Effects 0.000 description 2
- 229920000117 poly(dioxanone) Polymers 0.000 description 2
- 229920001553 poly(ethylene glycol)-block-polylactide methyl ether Polymers 0.000 description 2
- 239000012460 protein solution Substances 0.000 description 2
- 239000013557 residual solvent Substances 0.000 description 2
- 239000007962 solid dispersion Substances 0.000 description 2
- SCYULBFZEHDVBN-UHFFFAOYSA-N 1,1-Dichloroethane Chemical compound CC(Cl)Cl SCYULBFZEHDVBN-UHFFFAOYSA-N 0.000 description 1
- JJTUDXZGHPGLLC-IMJSIDKUSA-N 4511-42-6 Chemical compound C[C@@H]1OC(=O)[C@H](C)OC1=O JJTUDXZGHPGLLC-IMJSIDKUSA-N 0.000 description 1
- LPEKGGXMPWTOCB-UHFFFAOYSA-N 8beta-(2,3-epoxy-2-methylbutyryloxy)-14-acetoxytithifolin Natural products COC(=O)C(C)O LPEKGGXMPWTOCB-UHFFFAOYSA-N 0.000 description 1
- 229920000945 Amylopectin Polymers 0.000 description 1
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- RMOUBSOVHSONPZ-UHFFFAOYSA-N Isopropyl formate Chemical group CC(C)OC=O RMOUBSOVHSONPZ-UHFFFAOYSA-N 0.000 description 1
- 208000034530 PLAA-associated neurodevelopmental disease Diseases 0.000 description 1
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 1
- 229920001244 Poly(D,L-lactide) Polymers 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 229920002732 Polyanhydride Polymers 0.000 description 1
- 229920000432 Polylactide-block-poly(ethylene glycol)-block-polylactide Polymers 0.000 description 1
- 229920001710 Polyorthoester Polymers 0.000 description 1
- BCKXLBQYZLBQEK-KVVVOXFISA-M Sodium oleate Chemical compound [Na+].CCCCCCCC\C=C/CCCCCCCC([O-])=O BCKXLBQYZLBQEK-KVVVOXFISA-M 0.000 description 1
- 102000011923 Thyrotropin Human genes 0.000 description 1
- 108010061174 Thyrotropin Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000005215 alkyl ethers Chemical class 0.000 description 1
- REKYPYSUBKSCAT-UHFFFAOYSA-N beta-hydroxyvaleric acid Natural products CCC(O)CC(O)=O REKYPYSUBKSCAT-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 229940043232 butyl acetate Drugs 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 238000009295 crossflow filtration Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 229940116333 ethyl lactate Drugs 0.000 description 1
- 238000004880 explosion Methods 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 150000002191 fatty alcohols Chemical class 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- BWHLPLXXIDYSNW-UHFFFAOYSA-N ketorolac tromethamine Chemical compound OCC(N)(CO)CO.OC(=O)C1CCN2C1=CC=C2C(=O)C1=CC=CC=C1 BWHLPLXXIDYSNW-UHFFFAOYSA-N 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 229940057867 methyl lactate Drugs 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- YKYONYBAUNKHLG-UHFFFAOYSA-N n-Propyl acetate Natural products CCCOC(C)=O YKYONYBAUNKHLG-UHFFFAOYSA-N 0.000 description 1
- 239000002088 nanocapsule Substances 0.000 description 1
- 229920002113 octoxynol Polymers 0.000 description 1
- 239000008385 outer phase Substances 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- LQRJAEQXMSMEDP-XCHBZYMASA-N peptide a Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](C)C(=O)NCCCC[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)C(\NC(=O)[C@@H](CCCCN)NC(=O)CNC(C)=O)=C/C=1C=CC=CC=1)C(N)=O)C(=O)C(\NC(=O)[C@@H](CCCCN)NC(=O)CNC(C)=O)=C\C1=CC=CC=C1 LQRJAEQXMSMEDP-XCHBZYMASA-N 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 229920001983 poloxamer Polymers 0.000 description 1
- 229960000502 poloxamer Drugs 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229940068965 polysorbates Drugs 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 229940090181 propyl acetate Drugs 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
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- 210000002784 stomach Anatomy 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/38—Albumins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
- A61K38/09—Luteinising hormone-releasing hormone [LHRH], i.e. Gonadotropin-releasing hormone [GnRH]; Related peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/21—Interferons [IFN]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/28—Insulins
-
- 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)
-
- 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
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Abstract
The invention concerns a method of producing morphologically uniform microcapsules containing peptides, proteins or other water-soluble biologically active substances as active substance, and microcapsules produced by this method with a charging degree of between 3 and 30 wt.% and a diameter of <= 8 µm. According to the invention, biodegradable polymers are dissolved in a halogen-free solvent or solution mixture and the buffered active substance solution, which has a pH
of between 6.0 and 8.0, is dispersed in this solution. An aqueous solution containing a surfactant is then added to this W/O emulsion (W/O/W emulsion) and the solvent removed. The microcapsules produced by this method display no tendency to agglomerate. The encapsulation efficiency of the method is between 90 and 95 %.
of between 6.0 and 8.0, is dispersed in this solution. An aqueous solution containing a surfactant is then added to this W/O emulsion (W/O/W emulsion) and the solvent removed. The microcapsules produced by this method display no tendency to agglomerate. The encapsulation efficiency of the method is between 90 and 95 %.
Description
Process for the Production of Morphologically Uniform Microcapsules and Microcapsules that are Produced According to this Process The invention relates to a process for the production of morphologically uniform microcapsules that contain peptides, proteins, or other water-soluble biologically active substances as active ingredients, and microcapsules that are produced according to this process.
As is generally known, peptides and proteins represent active ingredients with sizeable pharmacodynamics, which, however, are broken down upon oral administration because of their hydrolysis sensitivity in the acidic environment of the stomach, as well as enzymatic degradation, and thus are partially inactivated in such a way that their action in the gastrointestinal tract is considerably reduced.
Rapid inactivation of proteins and peptides can be observed, however, even after parenteral administration and especially after intravenous administration because of the half-life, which is very often very short. This means that despite sizeable pharmacodynamics and theoretically lower therapeutic dosages, multiple administrations of higher dosages may be necessary, which mean a large burden on the patients.
Suitable formulations that avoid the above-mentioned drawbacks are depot systems in the form of polymer microcapsules or polymer nanocapsules, which are also known extensively for peptides and are described in the literature.
, CA 02238352 1998-OS-21
As is generally known, peptides and proteins represent active ingredients with sizeable pharmacodynamics, which, however, are broken down upon oral administration because of their hydrolysis sensitivity in the acidic environment of the stomach, as well as enzymatic degradation, and thus are partially inactivated in such a way that their action in the gastrointestinal tract is considerably reduced.
Rapid inactivation of proteins and peptides can be observed, however, even after parenteral administration and especially after intravenous administration because of the half-life, which is very often very short. This means that despite sizeable pharmacodynamics and theoretically lower therapeutic dosages, multiple administrations of higher dosages may be necessary, which mean a large burden on the patients.
Suitable formulations that avoid the above-mentioned drawbacks are depot systems in the form of polymer microcapsules or polymer nanocapsules, which are also known extensively for peptides and are described in the literature.
, CA 02238352 1998-OS-21
2 They have the advantages that -- Peptides and proteins are protected against rapid inactivation, -- lower dosages are pharmacologically effective, -- multiple administration can be reduced, -- controlled release of peptides and proteins is possible in principle, -- the encapsulated active ingredients are transported in a directed manner, and -- undesirable side-effects can be reduced.
The known processes for microencapsulation or nanoencapsulation of water-soluble substances can be divided as follows:
-- Coacervation or emulsion phase separation -- encapsulation by spray drying -- solvent-evaporation in an organic or aqueous phase.
All processes include the embedding of active ingredients into a biodegradable polymer matrix or copolymer matrix.
Polymers that are known from the literature for this purpose are polyamides, polyanhydrides, polyesters, polyorthoesters, polyacetates, polylactones, polyorthocarbonates, i.a. To date, polylactide-co-glycolide polymers have mainly been used.
Thus, pharmaceutical compositions of water-soluble peptides and proteins in capsule form, which were produced based on coacervation or emulsion phase separation, are known from US
4,675,189 (Syntex Inc.), US 4,835,139 (Debiopharm S.A.) and EP
302 582 B1 (Southern Research Inst.).
The known processes for microencapsulation or nanoencapsulation of water-soluble substances can be divided as follows:
-- Coacervation or emulsion phase separation -- encapsulation by spray drying -- solvent-evaporation in an organic or aqueous phase.
All processes include the embedding of active ingredients into a biodegradable polymer matrix or copolymer matrix.
Polymers that are known from the literature for this purpose are polyamides, polyanhydrides, polyesters, polyorthoesters, polyacetates, polylactones, polyorthocarbonates, i.a. To date, polylactide-co-glycolide polymers have mainly been used.
Thus, pharmaceutical compositions of water-soluble peptides and proteins in capsule form, which were produced based on coacervation or emulsion phase separation, are known from US
4,675,189 (Syntex Inc.), US 4,835,139 (Debiopharm S.A.) and EP
302 582 B1 (Southern Research Inst.).
3 According to this disclosure, processes are described in which the copolymer that is used, preferably poly-(lactide-co-glycolide)-polymer, is dissolved in a halogenated organic solvent, preferably dichloromethane, and an aqueous peptide solution is dispersed in this solution. Then, a so-called coacervation agent is added. The coacervation agent is soluble in the organic solvent, but the polymer is insoluble in the coacervation agent, causing precipitation of the polymer with the inclusion of the dispersed polypeptides. As a coacervation agent, usually silicone oil is used for phase separation. After the silicone oil is added, a large amount of heptane, which ensures the setting of the microcapsules, must be added as well.
The encapsulation efficiency of this method is approximately 70$ (US 4,835,136). The microcapsules that are produced have a diameter of 1 to 500 ~.cm, preferably 10 to 50 ~Cm according to the examples.
In addition to the use of toxicologically problematic agents such as dichloromethane, heptane, and silicone oil, the drawbacks of this process also include the need to use large amounts of solvent, which results from the encapsulation using coacervation agents, such as silicone oil.
A process that is described in EP-A 315875 (Hoechst AG) for the production of biodegradable microcapsules of water-soluble peptides and proteins is based on the spray-drying process, in which an aqueous peptide or protein solution is emulsified in an organic polymer solution, and this emulsion is spray-dried.
The encapsulation efficiency of this method is approximately 70$ (US 4,835,136). The microcapsules that are produced have a diameter of 1 to 500 ~.cm, preferably 10 to 50 ~Cm according to the examples.
In addition to the use of toxicologically problematic agents such as dichloromethane, heptane, and silicone oil, the drawbacks of this process also include the need to use large amounts of solvent, which results from the encapsulation using coacervation agents, such as silicone oil.
A process that is described in EP-A 315875 (Hoechst AG) for the production of biodegradable microcapsules of water-soluble peptides and proteins is based on the spray-drying process, in which an aqueous peptide or protein solution is emulsified in an organic polymer solution, and this emulsion is spray-dried.
4 As a biodegradable polymer, a mixture of polyhydroxybutyric acid and poly (lactide-co-glycolide) polymer is used in a mixing ratio of between 99:1 to 20:80.
The peptide or protein is present in micronized form or in an aqueous solution. As a solvent, chloroform, dichloromethane, DMF or a solvent mixture that consists of water/ethanol/chloroform are considered. According to the examples, chloroform is used. The spray drying is carried out at temperatures of between preferably 45 and 95°C.
Disadvantageous in this process is the potential risk of explosion when a non-halogenated solvent is used and high temperatures are used simultaneously during the drying process.
Moreover, the use of non-flammable solvents such as dichloroethane results in toxicologically harmful residual solvent contamination in the end product. In addition, spray-dried microcapsules basically show a strong tendency to agglomerate; agglomerates of about 100 E.cm in size are produced.
Microparticles that are produced according to the "solvent-evaporation-process'° are described in two Canadian Patent Applications CA 2,100,925 (Rhone-Merieux) and CA 2,099,941 (Tanabe Seiyaku Co.).
Usually, with this method, the aqueous peptide or protein solution is dispersed into an organic polymer solution, or active ingredient crystals are suspended in the polymer solution. After a second aqueous phase is added with a surfactant, the polymer solvent is evaporated.
This method is highly variable, and normally W/O- or complex W/O/W-emulsions are produced.
According to CA 2,099,941, water-soluble active ingredients and biodegradable polymers are first dissolved in a solvent or a solvent mixture in which they are both soluble. Then, this solvent is removed, and the solid dispersion that is produced is dissolved in an organic solvent that is not water-miscible. The resulting solution (oil phase) is emulsified in an aqueous phase, so that a W/O-emulsion is produced.
Finally, the organic solvent of the oil phase of this emulsion is evaporated.
Concrete examples of the patent relate to poly (lactide-co-glycolide) polymers (PLGA) as a matrix and a hormone (TRH) that releases thyreotropin or its derivatives as an active ingredient, which are first dissolved in a mixture that consists of acetonitrile/ethanol and optionally water, or only acetonitrile, or that consists of acetonitrile and aqueous gelatin, or of dichloromethane and ethanol.
As an organic solvent in the solution of the solid dispersion, dichloromethane or chloroform is used. An aqueous polyvinyl alcohol solution represents the aqueous phase.
The size of the microcapsules is approximately a diameter of 1 to 100 Vim, according to the concrete examples about 50 um to < 100 Vim.
According to CA 2,100,925, microcapsules of LHRH hormone and analogs are produced by prior dispersion of the LHRH hormone in powder form in two organic solvents, whereby one solvent (above-mentioned dispersion solvent) makes it possible to produce a homogeneous suspension of the pulverized hormone by simple stirring. The second solvent is readily water-miscible and thus makes the microdispersion of the organic phase in aqueous phase possible.
As a second solvent, dichloromethane or, alternatively, chloroform is used. The capsules have a diameter of between 1 and 250 E.cm. Preferably, the capsules are larger than 50-60 E.tm.
The morphology of the microcapsules that are thus produced is also very different. As already explained above, the halogenated solvents that are used are toxicologically harmful.
In addition, this process also requires sizeable amounts of surfactants.
The object of the invention was to develop a simple and gentle process for the production of morphologically uniform, non-agglomerating microcapsules using toxicologically harmless solvents, which have an encapsulation efficiency of at least 85~, preferably over 90~, and is to yield microcapsules in a size range of 200 nm to 500 ~Cm with a high degree of concentration.
In addition, the process is to make "scaling-up" possible.
The object of the invention is achieved surprisingly simply using the "Induced Phase Transition" method, which is implemented by virtue of the fact that a polymer that is commonly used for microcapsule production, such as a polyester that consists of hydroxycarboxylic acids or a block polymer that consists of hydroxycarboxylic acids and polyethylene glycol (PEG), is dissolved in a halogen-free, solvent or solvent mixture that is not water-miscible or is partially water-miscible and the buffered active ingredient solution, which has a pH of between 6.0 - 8.0, is dispersed in this solution. Homogenization produces a stable W/O-emulsion to which an aqueous solution that contains a surfactant or a mixture of surfactants is added as an outer phase while being stirred, in such a way that a three-phase W/O/W emulsion is obtained. Then, the solvent or solvent mixture .is removed with commonly used methods, preferably in a vacuum and/or air/nitrogen stream. The microcapsules are concentrated and optionally freeze-dried.
In this case, the particle size is controlled by the stirring speed, whereby smaller particles (< 8 Vim) -- such as are required if the product is intended for intravenous administration -- are obtained at higher stirring speeds.
Optionally, after the solvent is removed, the microcapsules are additionally subjected to "cross-flow" filtration, by which residual surfactant and residual solvent portions are removed.
As a result, it is possible to reduce or to avoid the "initial burst," i.e., a large release of active ingredients immediately after administration (because of active ingredients that adhere to the particle surface).
For freeze-drying, cryoprotectors such as sugar, sugar alcohols, or polyvinylpyrrolidone derivatives are optionally added.
Preferred polyesters of hydroxycarboxylic acids that can be used in the process according to the invention are:
. CA 02238352 1998-OS-21 Polyglycolides (PGA) and copolymers of glycolides, such as glycolide/lactide copolymers (PGA/PLLA) or glycolide/trimethylene carbonate copolymers (PGA/TMC); L-polylactides (PLA) and stereocopolymers of polylactides such as poly-L-lactide (PLLA), poly-DL-lactide copolymers and L-lactide/DL-lactide copolymers;
copolymers of PLA such as lactide/tetramethylglycolide copolymers, lactide/S-valerolactone copolymer and lactide/~-caprolactone copolymer; poly-f3-hydroxybutyrate (PHBA), PHBA/!3-hydroxyvalerate copolymers (PHBA/HVA), poly-f3-hydroxypropionate (PHPA), poly-p-dioxanone (PDS), poly-6-valerolactone, hydrophobized polysaccharides, - hyaluronic acid, dextrans or hydrophobized amylopectin and poly-~-caprolactone.
As block copolymers of polyesters of hydroxycarboxylic acids and linear or star-polyethylene glycol (PEG), the substances named below can be used in the process according to the invention:
AB-Block copolymers that consist of PLA and PEG, ABA-triblock copolymers that consist of PLA-PEG-PLA, S(3)-PEG-PLA
block copolymers and S(4)-PEG-PLA block copolymers.
The polymer Resomer~R~ 505, especially Resomer~R~ RG-756 or Resomer~R~ RG-858, is preferred according to the invention.
Resomer~R~ is a trademark of the Bohringer Ingelheim Company.
In this case, this is a (DL-lactide-co-glycolide)-polymer.
Halogen-free solvents or solvent mixtures that are preferred according to the invention are acetone, ethanol, alkyl acetates such as methyl, ethyl, propyl, isopropyl or butyl acetate, alkyl formates such as methyl-, ethyl-, propyl-, isopropyl- or butyl formate, triacetin, triethyl citrate and/or C~-C4 alkyl lactates, e.g., methyl or ethyl lactate.
Ethyl acetate, isopropyl acetate, and propyl formate are especially preferably used.
For the purposes of this invention, buffered solutions are aqueous solutions of peptides, proteins or their physiologically compatible salts or of other water-soluble biologically active substances, which are preferably adjusted with a tris (hydroxymethyl) aminomethane solution or a phosphate buffer solution to a pH of between 6.0 and 8.0, preferably a pH of 6.5 to 7.4.
Another buffer that can be used according to the invention is the citrate buffer, whereby the buffer concentrations are generally in the range of 5 mmol/1 to 300 mmol/1.
Any water-soluble peptides or proteins can be encapsulated with the process according to the invention. The process according to the invention is especially suitable for encapsulating human serum albumin, insulin, interferon, and LHRH
antagonists or their analogs.
Morphologically uniform microcapsules of human serum albumin, insulin, interferons, and the peptides that are mentioned below can quite especially advantageously be produced with the process according to the invention:
a) DesA(2)Nal-beta-Ala-DCpa-Deal-Ser-Tyr-DCit-Leu-Arg-Pro-DAla-NH2 , b) DesA(2)Nal-Gly-DCpa-DPal-Ser-Tyr-DCit-Leu-Arg-Pro-DAla-NHZ , c) Ac-DNal-DCpa-DPal-Ser-Tyr-DLys(Mor)-Leu-Lys(Mor)Pro-DAla-NHZ .
The meanings of DesA(2)Nal and D-Cit and the chemical structures of peptides a) to c) are presented in Figure 1 or 2.
For the purposes of the invention, preferred as surfactants are substances from the Poloxamere~R~ group, polyethylene glycol alkyl ethers, polysorbates (Tween~R~, Span~R~), saccharose esters (Sisterna~R~, The Netherlands), saccharose esters (Ryoto sugar esters, Tokyo), gelatin, polyvinylpyrrolidone, fatty alcohol polyglycoside, charps, charpso, decyl-f3-D-glycopyranoside, decyl-f3-D-maltopyranoside, dodecyl-f3-D-maltopyranoside, sodium-oleate, Poloxamine~R~ group, polyethylene glycol, polyvinyl alcohol, polyoxyethylated fatty acid ether (Brij~R~) Triton X 100 or mixtures thereof.
Polyvinyl alcohol, Brij~R~, Poloxamere~R~ Poloxamine~R~ and Tween~R~ are preferably used.
The subject of the invention is also morphologically uniform microcapsules that are produced according to the above-mentioned process and have a diameter of 200 nm to 500 Vim, preferably between 0.2 to 8 /..cm.
Because of the advantageous conformation of polymer and solvent, no formation of agglomerates of the microcapsules occurs in the process according to the invention.
Thus, Figures 3 and 4 show light-microscopic pictures of the microcapsules according to the invention that are produced according to Example 10 (Fig. 3) or according to Example 15 (Fig.
4). A millimeter in the imaging corresponds to 1 ~.Cm in reality.
The pictures clearly show the uniform morphology; particle agglomerates are not present.
The encapsulation efficiency of the process is at least 85~;
preferably encapsulation efficiencies of between 90 and 95~ are achieved. The mass of the encapsulated active ingredient 100/mass of the active ingredient that is used is defined as encapsulation efficiency. The degree of concentration of the microcapsules that are produced is between 3 to 30~ (degree of concentration = mass of active ingredient - 100/mass of active ingredient + mass of polymer).
Then, the invention is to be explained in more detail in the embodiments, without limiting the latter to it.
, CA 02238352 1998-OS-21 ' 12 Example 1 1.7 g of the polymer Resomer~R~ RG-756 is dissolved in 29 ml of ethyl acetate and moved into a steel vessel (height 11.5 cm, inside diameter 8 cm). Then, 3 ml of an aqueous 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.4) that contains 200 mg of human albumin is dispersed with the aid of a mechanical stirrer (Dispermat-FT, VMA-Getzmann GmbH, 5 cm dissolver disk) into the polymer solution for 6 minutes at 10,000 rpm below room temperature. 45 ml of an aqueous solution, consisting of a 2~
polyvinyl alcohol solution (molecular weight 9,000 - 10,000, Aldrich) is added to the W/O-emulsion that is produced while being stirred (8,000 rpm). After a dispersion time of 10 seconds, the W/O/W-emulsion is moved into a 500 ml three-necked flask and stirred using a KPG stirrer. The solvent ethyl acetate is then removed at 20°C by applying a vacuum (900 mbar), nitrogen, or air feed. After 5 hours, the suspension is washed with 5 1 of water or an aqueous solution and concentrated by evaporation in a desired suspension volume. "Cross-flow"
filtration is carried out using a Sartocon Mini~R~ (Sartorius AG, Gottingen) Systems. The solvent-free and almost emulsifier-free suspension is mixed with a cryoprotector (for example with a sugar, sugar alcohol, or polyvinyl pyrrolidone derivative), frozen as quickly as possible, for example with liquid nitrogen, and freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with a human albumin content of 9~ (human albumin mass - 100/human albumin mass +
polymer mass = degree of concentration), and they have a diameter of from 0.2 to 8 ~,m. The encapsulation efficiency is 86~.
Example 2 The procedure is the same as in Example 1, whereby 1.7 g of Resomer~R~ RG-756 is not dissolved in 29 ml of ethyl acetate, but rather in 40 ml of methyl acetate.
Example 3 The procedure is the same as in Example 1, whereby instead of 1.7 g of the polymer Resomer~R~ RG-756, 1.1 g of the polymer Resomer~R~ RG-858 is used.
Example 4 The procedure is the same as in Example 1, whereby instead of 1.7 g of the polymer resomer~R~ RG-756, 3.0 g of the polymer Resomer~R~ RG-858 is used.
Example 5 The procedure is the same as in Example 1, whereby instead of a 2~ PVA solution, a 2~ Brij~R~ 35 solution is used.
Example 6 The procedure is the same as in Example 1, whereby instead of a 2~ PVA solution, a 2~ Brij~R~ 96 solution is used.
Example 7 The procedure is the same as in Example 1, whereby instead of a 2~ PVA solution, a 2~ Tween~R~ 20 solution is used.
Example 8 1.1 g of the polymer Resomer~R~ RG-858 is dissolved in 29 ml of ethyl acetate and moved into a steel vessel (height 11.5 cm, inside diameter 8 cm).
Then, 7 ml of a 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.4) that contains 50 mg of the peptide DesA(2)Nal-beta-Ala-DCpa-deal-Ser-Tyr-DCit-Leu-Arg-Pro-DAla-NHZ (peptide a) and 2 ml of ethanol is dispersed with the aid of a mechanical stirrer (Dispermat-FT, VMA-Getzmann GmbH, 5 cm dissolver disk) into the polymer solution for 6 minutes at 10,000 rpm below room temperature. 45 ml of an aqueous solution, consisting of a 2~
polyvinyl alcohol solution (molecular weight 9,000 - 10,000, Aldrich) is added to the W/O-emulsion that is produced while being stirred (8,000 rpm). After a dispersion time of 10 seconds, the W/O/W-emulsion is moved into a 500 ml three-necked flask and stirred using a KPG stirrer. The solvent ethyl acetate is then removed at 20°C by applying a vacuum (900 mbar), nitrogen or air feed. After 5 hours, the suspension is washed with 5 1 of water or an aqueous solution and concentrated by evaporation in a desired suspension volume. A "cross-flow" filtration is carried out using a Sartocon Mini~R~ (Sartorius AG, Gottingen) System with a polyolefin membrane (cutoff 0.2 Vim). The solvent-free and almost emulsifier-free suspension is mixed with a cryoprotector (for example with a sugar, sugar alcohol or polyvinyl pyrrolidone derivative), frozen as quickly as possible, for example with liquid nitrogen, and freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with an active ingredient content of 4~. The microcapsules have a diameter of from 0.2 to 8 Vim. The encapsulation efficiency is 93~.
Example 9 1.1 g of the polymer Resomer~R~ RG-858 is dissolved in 29 ml of ethyl acetate and moved into a steel vessel (height 11.5 cm, inside diameter 8 cm). Then, 5 ml of an aqueous 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.4) that contains 48 mg of the peptide DesA(2)Nal-Gly-DCpa-DPal-Ser-Tyr-DCit-Leu-Arg-Pro-DAla-NHZ (peptide b) is dispersed with the aid of a mechanical stirrer (Dispermat-FT, VMA-Getzmann GmbH, 5 cm dissolver disk) into the polymer solution for 6 minutes at 10,000 rpm below room temperature. 45 ml of an aqueous solution, consisting of a 2~ polyvinyl alcohol solution (molecular weight 9,000 - 10,000, Aldrich) is added to the W/O-emulsion that is produced while being stirred (8,000 rpm). After a dispersion time of 10 seconds, the W/O/W-emulsion is moved into a 500 ml three-necked flask and stirred using a KPG stirrer. The solvent ethyl acetate is then removed at 20°C by applying a vacuum (900 mbar), nitrogen or air feed. After 5 hours, the suspension is washed with 5 1 of water or an aqueous solution and concentrated by evaporation in a desired suspension volume. The use of a "cross-flow°° filtration, for example with a Sartocon Mini~R~
(Sartorius AG, Gottingen) System with a polyolefin membrane (cutoff 0.2 Vim), is advantageous. The solvent-free and almost emulsifier-free suspension can be mixed with a cryoprotector (for example with a sugar, sugar alcohol or polyvinyl pyrrolidone derivative) and is frozen as quickly as possible, for example with liquid nitrogen, and freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with an active ingredient content of 4$. The microcapsules have a diameter of from 0.2 to 8 ~tcm. The encapsulation efficiency is 95.7.
Example 10 1.1 g of the polymer Resomer~R~ RG-858 is dissolved in 30 ml of propyl formate and moved into a steel vessel (height 11.5 cm, inside diameter 8 cm). Then, 5 ml of an aqueous 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.0) that contains 50 mg of the LHRH antagonist Ac-DNal-DCpa-DPal-Ser-Tyr-DLys (Mor)-Leu-Lys(Mor)Pro-DAla-NH2 (peptide c) is dispersed with the aid of a mechanical stirrer (Dispermat-FT, VMA-Getzmann GmbH, 5 cm dissolver disk) into the polymer solution for 6 minutes at 10,000 rpm below room temperature. 45 ml of an aqueous solution, consisting of a 2~ polyvinyl alcohol solution (molecular weight x,000 - 10,000, Aldrich), is added to the W/O-emulsion that is produced while being stirred (8,000 rpm). After a dispersion time of 10 seconds, the W/O/W-emulsion is moved into a 500 ml three-necked flask and stirred using a KPG stirrer. The solvent propyl formate is then removed at 20°C by applying a vacuum (900 mbar), nitrogen or air feed. After 5 hours, the suspension is washed with 5 1 of water or an aqueous solution and concentrated by evaporation in a desired suspension volume. A "cross-flow"
filtration is carried out with a Sartocon Mini~R~ (Sartorius AG, Gottingen) System with a polyolefin membrane (cutoff 0.2 um).
The solvent-free and almost emulsifier-free suspension is frozen as quickly as possible with liquid nitrogen and freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with an active ingredient content of 3.9~, and the microcapsules have a diameter of from 0.2 to 8 Vim. The encapsulation efficiency is 90.7.
Example 11 1.1 g of the polymer Resomer~R~ RG-858 is dissolved in 30 ml of isopropyl acetate and moved into a steel vessel (height 11.5 cm, inside diameter 8 cm). Then, 5 ml of an aqueous 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.0) that contains 50 mg of the LHRH antagonist as in Example 10 is dispersed with the aid of a mechanical stirrer (Dispermat-FT, VMA-Getzmann GmbH, cm dissolver disk) into the polymer solution for 6 minutes at 10,000 rpm below room temperature.
45 ml of an aqueous solution, consisting of a 2~ polyvinyl alcohol solution (molecular weight 9,000 - 10,000, Aldrich) is added to the W/O-emulsion that is produced while being stirred (8,000 rpm). After a dispersion time of 10 seconds, the W/O/W-emulsion is moved into a 500 ml three-necked flask and stirred 1 ti using a KPG stirrer. The solvent isopropyl acetate is then removed at 20°C by applying a vacuum (900 mbar), nitrogen or air feed. After 5 hours, the suspension is washed with 5 1 of water or an aqueous solution and concentrated by evaporation in a desired suspension volume. A "cross-flow" filtration is carried out with a Sartocon Mini~R~ (Sartorius AG, Gottingen) System with a polyolefin membrane (cutoff 0.2 ~Cm), and the solvent-free and almost emulsifier-free suspension is freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with an active ingredient content of 2.9~, and the microcapsules have a diameter of from 0.2 to 8 ~.im. The encapsulation efficiency is 90.6.
Example 12 The procedure is the same as in Example 1, whereby the 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.0) is replaced by a 5 mmol phosphate buffer solution (PBS, pH 7.2).
Example 13 The procedure is the same as in Example 1, whereby instead of 200 mg of HSA that is dissolved in 3 ml of tris-buffer (pH =
7.4), 750 mg of HSA that is dissolved in 5 ml of tris-buffer (pH
- 7.4) is used.
The lyophilizate that is resuspended in water or aqueous solutions contains microcapsules with an HSA content of 30~. The encapsulation efficiency is 90.9.
' 19 Example 14 The procedure is the same as in Example 13, whereby instead of 2~ polyvinyl alcohol solution, a 2~ Poloxamer F 127 solution is used.
Example 15 The procedure is the same as in Example 13, whereby instead of 2~ polyvinyl alcohol solution, a 2~ Poloxamine T 707 solution is used.
Example 16 The procedure is the same as in Example 13, whereby instead of 2~ polyvinyl alcohol solution, a 2~ Poloxamine T 908 solution is used.
Example 17 The procedure is the same as in Example l, whereby 200 mg of HSA is replaced by 200 mg of insulin (human, recombinant (pfs), Sigma Chemie [Sigma Chemistry) No. I 0259).
Example 18 The procedure is the same as in Example l, whereby 200 mg of HSA is replaced by 200 mg of interferon (human leukocyte (pfs) (a-IFH, Le), Sigma Chemie No. I 1008).
~u Example 19 The procedure is the same as in Example 1, whereby 200 mg of HSA is replaced by 200 mg of insulin (human, gamma (pfs) (y-IFN), Sigma Chemie No. I 6507).
Example 20 120 mg of insulin is dissolved in 0.8 ml of HC1 (O.iN) and mixed with 2 ml of NaCl solution (0.9~). Then, the pH of the solution is set at 6-7 with NaOH (0.1N). This solution is added to a solution of 500 mg of polymer RG-858 in 10 ml of ethyl acetate and then stirred with Ultraturax (at 10,000-15,000 rpm) for 3-4 minutes. Then, 50 ml of a 2~ aqueous Polaxamine T707 solution is added while being stirred. After the addition has been completed, the suspension is moved into a three-necked flask, which is equipped with a stirrer. The solvent (ethyl acetate) is removed by applying a vacuum while being stirred.
The remaining residue is washed in a cross-flow filtration (Sartocon Mini~R~ Sartorius AG, Gottingen) with 5 liters of water.
The residue that contains nearly solvent-free and surfactant-free microcapsules is, optionally with the addition of a cryoprotector, quickly frozen and freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with a degree of concentration of 15~ by weight.
Example 21 The procedure is the same as in Example 20, whereby instead of the 2~ Polaxamine T707 solution, a 2~ Polaxamer-407 (F127) solution is used.
The degree of concentration of the microcapsules that are obtained is 17~.
Example 22 The procedure is the same as in Example 20, whereby instead of the 2$ Polaxamine T707 solution, a 2~ Polaxamer-188 (F68) solution is used.
The degree of concentration of the microcapsules that are obtained is 16~.
Example 23 The procedure is performed analogously to Example 20. Here, however, 700 mg of polymer and 223 mg of insulin are used. The lyophilizate that is resuspended in water or an aqueous solution contains microcapsules with an insulin content of 20~. -
The peptide or protein is present in micronized form or in an aqueous solution. As a solvent, chloroform, dichloromethane, DMF or a solvent mixture that consists of water/ethanol/chloroform are considered. According to the examples, chloroform is used. The spray drying is carried out at temperatures of between preferably 45 and 95°C.
Disadvantageous in this process is the potential risk of explosion when a non-halogenated solvent is used and high temperatures are used simultaneously during the drying process.
Moreover, the use of non-flammable solvents such as dichloroethane results in toxicologically harmful residual solvent contamination in the end product. In addition, spray-dried microcapsules basically show a strong tendency to agglomerate; agglomerates of about 100 E.cm in size are produced.
Microparticles that are produced according to the "solvent-evaporation-process'° are described in two Canadian Patent Applications CA 2,100,925 (Rhone-Merieux) and CA 2,099,941 (Tanabe Seiyaku Co.).
Usually, with this method, the aqueous peptide or protein solution is dispersed into an organic polymer solution, or active ingredient crystals are suspended in the polymer solution. After a second aqueous phase is added with a surfactant, the polymer solvent is evaporated.
This method is highly variable, and normally W/O- or complex W/O/W-emulsions are produced.
According to CA 2,099,941, water-soluble active ingredients and biodegradable polymers are first dissolved in a solvent or a solvent mixture in which they are both soluble. Then, this solvent is removed, and the solid dispersion that is produced is dissolved in an organic solvent that is not water-miscible. The resulting solution (oil phase) is emulsified in an aqueous phase, so that a W/O-emulsion is produced.
Finally, the organic solvent of the oil phase of this emulsion is evaporated.
Concrete examples of the patent relate to poly (lactide-co-glycolide) polymers (PLGA) as a matrix and a hormone (TRH) that releases thyreotropin or its derivatives as an active ingredient, which are first dissolved in a mixture that consists of acetonitrile/ethanol and optionally water, or only acetonitrile, or that consists of acetonitrile and aqueous gelatin, or of dichloromethane and ethanol.
As an organic solvent in the solution of the solid dispersion, dichloromethane or chloroform is used. An aqueous polyvinyl alcohol solution represents the aqueous phase.
The size of the microcapsules is approximately a diameter of 1 to 100 Vim, according to the concrete examples about 50 um to < 100 Vim.
According to CA 2,100,925, microcapsules of LHRH hormone and analogs are produced by prior dispersion of the LHRH hormone in powder form in two organic solvents, whereby one solvent (above-mentioned dispersion solvent) makes it possible to produce a homogeneous suspension of the pulverized hormone by simple stirring. The second solvent is readily water-miscible and thus makes the microdispersion of the organic phase in aqueous phase possible.
As a second solvent, dichloromethane or, alternatively, chloroform is used. The capsules have a diameter of between 1 and 250 E.cm. Preferably, the capsules are larger than 50-60 E.tm.
The morphology of the microcapsules that are thus produced is also very different. As already explained above, the halogenated solvents that are used are toxicologically harmful.
In addition, this process also requires sizeable amounts of surfactants.
The object of the invention was to develop a simple and gentle process for the production of morphologically uniform, non-agglomerating microcapsules using toxicologically harmless solvents, which have an encapsulation efficiency of at least 85~, preferably over 90~, and is to yield microcapsules in a size range of 200 nm to 500 ~Cm with a high degree of concentration.
In addition, the process is to make "scaling-up" possible.
The object of the invention is achieved surprisingly simply using the "Induced Phase Transition" method, which is implemented by virtue of the fact that a polymer that is commonly used for microcapsule production, such as a polyester that consists of hydroxycarboxylic acids or a block polymer that consists of hydroxycarboxylic acids and polyethylene glycol (PEG), is dissolved in a halogen-free, solvent or solvent mixture that is not water-miscible or is partially water-miscible and the buffered active ingredient solution, which has a pH of between 6.0 - 8.0, is dispersed in this solution. Homogenization produces a stable W/O-emulsion to which an aqueous solution that contains a surfactant or a mixture of surfactants is added as an outer phase while being stirred, in such a way that a three-phase W/O/W emulsion is obtained. Then, the solvent or solvent mixture .is removed with commonly used methods, preferably in a vacuum and/or air/nitrogen stream. The microcapsules are concentrated and optionally freeze-dried.
In this case, the particle size is controlled by the stirring speed, whereby smaller particles (< 8 Vim) -- such as are required if the product is intended for intravenous administration -- are obtained at higher stirring speeds.
Optionally, after the solvent is removed, the microcapsules are additionally subjected to "cross-flow" filtration, by which residual surfactant and residual solvent portions are removed.
As a result, it is possible to reduce or to avoid the "initial burst," i.e., a large release of active ingredients immediately after administration (because of active ingredients that adhere to the particle surface).
For freeze-drying, cryoprotectors such as sugar, sugar alcohols, or polyvinylpyrrolidone derivatives are optionally added.
Preferred polyesters of hydroxycarboxylic acids that can be used in the process according to the invention are:
. CA 02238352 1998-OS-21 Polyglycolides (PGA) and copolymers of glycolides, such as glycolide/lactide copolymers (PGA/PLLA) or glycolide/trimethylene carbonate copolymers (PGA/TMC); L-polylactides (PLA) and stereocopolymers of polylactides such as poly-L-lactide (PLLA), poly-DL-lactide copolymers and L-lactide/DL-lactide copolymers;
copolymers of PLA such as lactide/tetramethylglycolide copolymers, lactide/S-valerolactone copolymer and lactide/~-caprolactone copolymer; poly-f3-hydroxybutyrate (PHBA), PHBA/!3-hydroxyvalerate copolymers (PHBA/HVA), poly-f3-hydroxypropionate (PHPA), poly-p-dioxanone (PDS), poly-6-valerolactone, hydrophobized polysaccharides, - hyaluronic acid, dextrans or hydrophobized amylopectin and poly-~-caprolactone.
As block copolymers of polyesters of hydroxycarboxylic acids and linear or star-polyethylene glycol (PEG), the substances named below can be used in the process according to the invention:
AB-Block copolymers that consist of PLA and PEG, ABA-triblock copolymers that consist of PLA-PEG-PLA, S(3)-PEG-PLA
block copolymers and S(4)-PEG-PLA block copolymers.
The polymer Resomer~R~ 505, especially Resomer~R~ RG-756 or Resomer~R~ RG-858, is preferred according to the invention.
Resomer~R~ is a trademark of the Bohringer Ingelheim Company.
In this case, this is a (DL-lactide-co-glycolide)-polymer.
Halogen-free solvents or solvent mixtures that are preferred according to the invention are acetone, ethanol, alkyl acetates such as methyl, ethyl, propyl, isopropyl or butyl acetate, alkyl formates such as methyl-, ethyl-, propyl-, isopropyl- or butyl formate, triacetin, triethyl citrate and/or C~-C4 alkyl lactates, e.g., methyl or ethyl lactate.
Ethyl acetate, isopropyl acetate, and propyl formate are especially preferably used.
For the purposes of this invention, buffered solutions are aqueous solutions of peptides, proteins or their physiologically compatible salts or of other water-soluble biologically active substances, which are preferably adjusted with a tris (hydroxymethyl) aminomethane solution or a phosphate buffer solution to a pH of between 6.0 and 8.0, preferably a pH of 6.5 to 7.4.
Another buffer that can be used according to the invention is the citrate buffer, whereby the buffer concentrations are generally in the range of 5 mmol/1 to 300 mmol/1.
Any water-soluble peptides or proteins can be encapsulated with the process according to the invention. The process according to the invention is especially suitable for encapsulating human serum albumin, insulin, interferon, and LHRH
antagonists or their analogs.
Morphologically uniform microcapsules of human serum albumin, insulin, interferons, and the peptides that are mentioned below can quite especially advantageously be produced with the process according to the invention:
a) DesA(2)Nal-beta-Ala-DCpa-Deal-Ser-Tyr-DCit-Leu-Arg-Pro-DAla-NH2 , b) DesA(2)Nal-Gly-DCpa-DPal-Ser-Tyr-DCit-Leu-Arg-Pro-DAla-NHZ , c) Ac-DNal-DCpa-DPal-Ser-Tyr-DLys(Mor)-Leu-Lys(Mor)Pro-DAla-NHZ .
The meanings of DesA(2)Nal and D-Cit and the chemical structures of peptides a) to c) are presented in Figure 1 or 2.
For the purposes of the invention, preferred as surfactants are substances from the Poloxamere~R~ group, polyethylene glycol alkyl ethers, polysorbates (Tween~R~, Span~R~), saccharose esters (Sisterna~R~, The Netherlands), saccharose esters (Ryoto sugar esters, Tokyo), gelatin, polyvinylpyrrolidone, fatty alcohol polyglycoside, charps, charpso, decyl-f3-D-glycopyranoside, decyl-f3-D-maltopyranoside, dodecyl-f3-D-maltopyranoside, sodium-oleate, Poloxamine~R~ group, polyethylene glycol, polyvinyl alcohol, polyoxyethylated fatty acid ether (Brij~R~) Triton X 100 or mixtures thereof.
Polyvinyl alcohol, Brij~R~, Poloxamere~R~ Poloxamine~R~ and Tween~R~ are preferably used.
The subject of the invention is also morphologically uniform microcapsules that are produced according to the above-mentioned process and have a diameter of 200 nm to 500 Vim, preferably between 0.2 to 8 /..cm.
Because of the advantageous conformation of polymer and solvent, no formation of agglomerates of the microcapsules occurs in the process according to the invention.
Thus, Figures 3 and 4 show light-microscopic pictures of the microcapsules according to the invention that are produced according to Example 10 (Fig. 3) or according to Example 15 (Fig.
4). A millimeter in the imaging corresponds to 1 ~.Cm in reality.
The pictures clearly show the uniform morphology; particle agglomerates are not present.
The encapsulation efficiency of the process is at least 85~;
preferably encapsulation efficiencies of between 90 and 95~ are achieved. The mass of the encapsulated active ingredient 100/mass of the active ingredient that is used is defined as encapsulation efficiency. The degree of concentration of the microcapsules that are produced is between 3 to 30~ (degree of concentration = mass of active ingredient - 100/mass of active ingredient + mass of polymer).
Then, the invention is to be explained in more detail in the embodiments, without limiting the latter to it.
, CA 02238352 1998-OS-21 ' 12 Example 1 1.7 g of the polymer Resomer~R~ RG-756 is dissolved in 29 ml of ethyl acetate and moved into a steel vessel (height 11.5 cm, inside diameter 8 cm). Then, 3 ml of an aqueous 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.4) that contains 200 mg of human albumin is dispersed with the aid of a mechanical stirrer (Dispermat-FT, VMA-Getzmann GmbH, 5 cm dissolver disk) into the polymer solution for 6 minutes at 10,000 rpm below room temperature. 45 ml of an aqueous solution, consisting of a 2~
polyvinyl alcohol solution (molecular weight 9,000 - 10,000, Aldrich) is added to the W/O-emulsion that is produced while being stirred (8,000 rpm). After a dispersion time of 10 seconds, the W/O/W-emulsion is moved into a 500 ml three-necked flask and stirred using a KPG stirrer. The solvent ethyl acetate is then removed at 20°C by applying a vacuum (900 mbar), nitrogen, or air feed. After 5 hours, the suspension is washed with 5 1 of water or an aqueous solution and concentrated by evaporation in a desired suspension volume. "Cross-flow"
filtration is carried out using a Sartocon Mini~R~ (Sartorius AG, Gottingen) Systems. The solvent-free and almost emulsifier-free suspension is mixed with a cryoprotector (for example with a sugar, sugar alcohol, or polyvinyl pyrrolidone derivative), frozen as quickly as possible, for example with liquid nitrogen, and freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with a human albumin content of 9~ (human albumin mass - 100/human albumin mass +
polymer mass = degree of concentration), and they have a diameter of from 0.2 to 8 ~,m. The encapsulation efficiency is 86~.
Example 2 The procedure is the same as in Example 1, whereby 1.7 g of Resomer~R~ RG-756 is not dissolved in 29 ml of ethyl acetate, but rather in 40 ml of methyl acetate.
Example 3 The procedure is the same as in Example 1, whereby instead of 1.7 g of the polymer Resomer~R~ RG-756, 1.1 g of the polymer Resomer~R~ RG-858 is used.
Example 4 The procedure is the same as in Example 1, whereby instead of 1.7 g of the polymer resomer~R~ RG-756, 3.0 g of the polymer Resomer~R~ RG-858 is used.
Example 5 The procedure is the same as in Example 1, whereby instead of a 2~ PVA solution, a 2~ Brij~R~ 35 solution is used.
Example 6 The procedure is the same as in Example 1, whereby instead of a 2~ PVA solution, a 2~ Brij~R~ 96 solution is used.
Example 7 The procedure is the same as in Example 1, whereby instead of a 2~ PVA solution, a 2~ Tween~R~ 20 solution is used.
Example 8 1.1 g of the polymer Resomer~R~ RG-858 is dissolved in 29 ml of ethyl acetate and moved into a steel vessel (height 11.5 cm, inside diameter 8 cm).
Then, 7 ml of a 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.4) that contains 50 mg of the peptide DesA(2)Nal-beta-Ala-DCpa-deal-Ser-Tyr-DCit-Leu-Arg-Pro-DAla-NHZ (peptide a) and 2 ml of ethanol is dispersed with the aid of a mechanical stirrer (Dispermat-FT, VMA-Getzmann GmbH, 5 cm dissolver disk) into the polymer solution for 6 minutes at 10,000 rpm below room temperature. 45 ml of an aqueous solution, consisting of a 2~
polyvinyl alcohol solution (molecular weight 9,000 - 10,000, Aldrich) is added to the W/O-emulsion that is produced while being stirred (8,000 rpm). After a dispersion time of 10 seconds, the W/O/W-emulsion is moved into a 500 ml three-necked flask and stirred using a KPG stirrer. The solvent ethyl acetate is then removed at 20°C by applying a vacuum (900 mbar), nitrogen or air feed. After 5 hours, the suspension is washed with 5 1 of water or an aqueous solution and concentrated by evaporation in a desired suspension volume. A "cross-flow" filtration is carried out using a Sartocon Mini~R~ (Sartorius AG, Gottingen) System with a polyolefin membrane (cutoff 0.2 Vim). The solvent-free and almost emulsifier-free suspension is mixed with a cryoprotector (for example with a sugar, sugar alcohol or polyvinyl pyrrolidone derivative), frozen as quickly as possible, for example with liquid nitrogen, and freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with an active ingredient content of 4~. The microcapsules have a diameter of from 0.2 to 8 Vim. The encapsulation efficiency is 93~.
Example 9 1.1 g of the polymer Resomer~R~ RG-858 is dissolved in 29 ml of ethyl acetate and moved into a steel vessel (height 11.5 cm, inside diameter 8 cm). Then, 5 ml of an aqueous 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.4) that contains 48 mg of the peptide DesA(2)Nal-Gly-DCpa-DPal-Ser-Tyr-DCit-Leu-Arg-Pro-DAla-NHZ (peptide b) is dispersed with the aid of a mechanical stirrer (Dispermat-FT, VMA-Getzmann GmbH, 5 cm dissolver disk) into the polymer solution for 6 minutes at 10,000 rpm below room temperature. 45 ml of an aqueous solution, consisting of a 2~ polyvinyl alcohol solution (molecular weight 9,000 - 10,000, Aldrich) is added to the W/O-emulsion that is produced while being stirred (8,000 rpm). After a dispersion time of 10 seconds, the W/O/W-emulsion is moved into a 500 ml three-necked flask and stirred using a KPG stirrer. The solvent ethyl acetate is then removed at 20°C by applying a vacuum (900 mbar), nitrogen or air feed. After 5 hours, the suspension is washed with 5 1 of water or an aqueous solution and concentrated by evaporation in a desired suspension volume. The use of a "cross-flow°° filtration, for example with a Sartocon Mini~R~
(Sartorius AG, Gottingen) System with a polyolefin membrane (cutoff 0.2 Vim), is advantageous. The solvent-free and almost emulsifier-free suspension can be mixed with a cryoprotector (for example with a sugar, sugar alcohol or polyvinyl pyrrolidone derivative) and is frozen as quickly as possible, for example with liquid nitrogen, and freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with an active ingredient content of 4$. The microcapsules have a diameter of from 0.2 to 8 ~tcm. The encapsulation efficiency is 95.7.
Example 10 1.1 g of the polymer Resomer~R~ RG-858 is dissolved in 30 ml of propyl formate and moved into a steel vessel (height 11.5 cm, inside diameter 8 cm). Then, 5 ml of an aqueous 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.0) that contains 50 mg of the LHRH antagonist Ac-DNal-DCpa-DPal-Ser-Tyr-DLys (Mor)-Leu-Lys(Mor)Pro-DAla-NH2 (peptide c) is dispersed with the aid of a mechanical stirrer (Dispermat-FT, VMA-Getzmann GmbH, 5 cm dissolver disk) into the polymer solution for 6 minutes at 10,000 rpm below room temperature. 45 ml of an aqueous solution, consisting of a 2~ polyvinyl alcohol solution (molecular weight x,000 - 10,000, Aldrich), is added to the W/O-emulsion that is produced while being stirred (8,000 rpm). After a dispersion time of 10 seconds, the W/O/W-emulsion is moved into a 500 ml three-necked flask and stirred using a KPG stirrer. The solvent propyl formate is then removed at 20°C by applying a vacuum (900 mbar), nitrogen or air feed. After 5 hours, the suspension is washed with 5 1 of water or an aqueous solution and concentrated by evaporation in a desired suspension volume. A "cross-flow"
filtration is carried out with a Sartocon Mini~R~ (Sartorius AG, Gottingen) System with a polyolefin membrane (cutoff 0.2 um).
The solvent-free and almost emulsifier-free suspension is frozen as quickly as possible with liquid nitrogen and freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with an active ingredient content of 3.9~, and the microcapsules have a diameter of from 0.2 to 8 Vim. The encapsulation efficiency is 90.7.
Example 11 1.1 g of the polymer Resomer~R~ RG-858 is dissolved in 30 ml of isopropyl acetate and moved into a steel vessel (height 11.5 cm, inside diameter 8 cm). Then, 5 ml of an aqueous 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.0) that contains 50 mg of the LHRH antagonist as in Example 10 is dispersed with the aid of a mechanical stirrer (Dispermat-FT, VMA-Getzmann GmbH, cm dissolver disk) into the polymer solution for 6 minutes at 10,000 rpm below room temperature.
45 ml of an aqueous solution, consisting of a 2~ polyvinyl alcohol solution (molecular weight 9,000 - 10,000, Aldrich) is added to the W/O-emulsion that is produced while being stirred (8,000 rpm). After a dispersion time of 10 seconds, the W/O/W-emulsion is moved into a 500 ml three-necked flask and stirred 1 ti using a KPG stirrer. The solvent isopropyl acetate is then removed at 20°C by applying a vacuum (900 mbar), nitrogen or air feed. After 5 hours, the suspension is washed with 5 1 of water or an aqueous solution and concentrated by evaporation in a desired suspension volume. A "cross-flow" filtration is carried out with a Sartocon Mini~R~ (Sartorius AG, Gottingen) System with a polyolefin membrane (cutoff 0.2 ~Cm), and the solvent-free and almost emulsifier-free suspension is freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with an active ingredient content of 2.9~, and the microcapsules have a diameter of from 0.2 to 8 ~.im. The encapsulation efficiency is 90.6.
Example 12 The procedure is the same as in Example 1, whereby the 5 mmol tris(hydroxymethyl)aminomethane solution (pH 7.0) is replaced by a 5 mmol phosphate buffer solution (PBS, pH 7.2).
Example 13 The procedure is the same as in Example 1, whereby instead of 200 mg of HSA that is dissolved in 3 ml of tris-buffer (pH =
7.4), 750 mg of HSA that is dissolved in 5 ml of tris-buffer (pH
- 7.4) is used.
The lyophilizate that is resuspended in water or aqueous solutions contains microcapsules with an HSA content of 30~. The encapsulation efficiency is 90.9.
' 19 Example 14 The procedure is the same as in Example 13, whereby instead of 2~ polyvinyl alcohol solution, a 2~ Poloxamer F 127 solution is used.
Example 15 The procedure is the same as in Example 13, whereby instead of 2~ polyvinyl alcohol solution, a 2~ Poloxamine T 707 solution is used.
Example 16 The procedure is the same as in Example 13, whereby instead of 2~ polyvinyl alcohol solution, a 2~ Poloxamine T 908 solution is used.
Example 17 The procedure is the same as in Example l, whereby 200 mg of HSA is replaced by 200 mg of insulin (human, recombinant (pfs), Sigma Chemie [Sigma Chemistry) No. I 0259).
Example 18 The procedure is the same as in Example l, whereby 200 mg of HSA is replaced by 200 mg of interferon (human leukocyte (pfs) (a-IFH, Le), Sigma Chemie No. I 1008).
~u Example 19 The procedure is the same as in Example 1, whereby 200 mg of HSA is replaced by 200 mg of insulin (human, gamma (pfs) (y-IFN), Sigma Chemie No. I 6507).
Example 20 120 mg of insulin is dissolved in 0.8 ml of HC1 (O.iN) and mixed with 2 ml of NaCl solution (0.9~). Then, the pH of the solution is set at 6-7 with NaOH (0.1N). This solution is added to a solution of 500 mg of polymer RG-858 in 10 ml of ethyl acetate and then stirred with Ultraturax (at 10,000-15,000 rpm) for 3-4 minutes. Then, 50 ml of a 2~ aqueous Polaxamine T707 solution is added while being stirred. After the addition has been completed, the suspension is moved into a three-necked flask, which is equipped with a stirrer. The solvent (ethyl acetate) is removed by applying a vacuum while being stirred.
The remaining residue is washed in a cross-flow filtration (Sartocon Mini~R~ Sartorius AG, Gottingen) with 5 liters of water.
The residue that contains nearly solvent-free and surfactant-free microcapsules is, optionally with the addition of a cryoprotector, quickly frozen and freeze-dried.
The lyophilizate that is resuspended with water or with an aqueous solution contains microcapsules with a degree of concentration of 15~ by weight.
Example 21 The procedure is the same as in Example 20, whereby instead of the 2~ Polaxamine T707 solution, a 2~ Polaxamer-407 (F127) solution is used.
The degree of concentration of the microcapsules that are obtained is 17~.
Example 22 The procedure is the same as in Example 20, whereby instead of the 2$ Polaxamine T707 solution, a 2~ Polaxamer-188 (F68) solution is used.
The degree of concentration of the microcapsules that are obtained is 16~.
Example 23 The procedure is performed analogously to Example 20. Here, however, 700 mg of polymer and 223 mg of insulin are used. The lyophilizate that is resuspended in water or an aqueous solution contains microcapsules with an insulin content of 20~. -
Claims (12)
1. Process for the production of morphologically uniform microcapsules that consist of biodegradable polymers or copolymers, which contain peptides, proteins or other water-soluble, biologically active substances as active ingredients, characterized in that -- Polyesters of hydroxycarboxylic acids or block copolymers that consist of polyesters of hydroxycarboxylic acids and polyethylene glycol are dissolved in a halogen-free solvent or solvent mixture that is not water-miscible or is partially water-miscible, -- a buffered active ingredient solution, which has a pH
of between 6.0 to 8.0, is dispersed into this solution, -- an aqueous solution that contains a surface-active substance or a mixture of surface-active substances is then added to this W/O-emulsion, and -- finally, the solvent or solvent mixture is removed in the usual way.
of between 6.0 to 8.0, is dispersed into this solution, -- an aqueous solution that contains a surface-active substance or a mixture of surface-active substances is then added to this W/O-emulsion, and -- finally, the solvent or solvent mixture is removed in the usual way.
2. Process according to claim 1, wherein as halogen-free solvents, acetone, ethanol; C1-C4 alkyl acetates, triacetin, triethyl citrate, C1-C4 alkyl formates and C1-C4 alkyl lactates or mixtures thereof are used.
3. Process according to one of claims 1 or 2, wherein as halogen-free solvents, methyl acetate, ethyl acetate, isopropyl acetate and propyl formate are used.
4. Process according to one of claims 1 to 3, wherein the buffered active ingredient solution comprises a phosphate buffer solution, a citrate buffer solution or a tris(hydroxymethyl)aminomethane solution is used.
5. Process according to one of claims 1 to 4, wherein the buffered active ingredient solution has a pH of between 6.0 to 8Ø
6. Process according to one of claims 1 to 4, wherein a polymer, Resomer(R) is used.
7. Process according to claim 6, wherein the polymer, Resomer(R) RG-756 or Resomer(R) RG-858, is used.
8. Process according to one of claims 1 to 7, wherein as active ingredients, human serum albumin, peptides, proteins, interferons, Betaseron(R), insulin, LHRH
antagonists or analogs thereof are used.
antagonists or analogs thereof are used.
9. Process according to one of claims 1 to 8, wherein as active ingredients a) DesA(2)Nal-beta Ala-DCpa-DPal-Ser-Tyr-DCit-Leu-Arg-Pro-DAla-NH2, b) DesA(2)Nal-G1y-DCpa-Deal-Ser-Tyr-DCit-Leu-Arg-Pro-DAla-NH-2 or c) Ac-DNal-DCpa-Deal-Ser-Tyr-DLys(Mor)-Leu-Lys(Mor)Pro-DAla-NH2 are used.
10. Process according to one of claims 1 to 9, wherein the removal of the solvent or solvent mixture, as well as the active ingredient and/or surfactant, is carried out by filtration.
11. Morphologically uniform microcapsules with a degree of concentration of between 3 to 30% by weight and a diameter of 200 nm to 500 µm produced according to the process of any one of claims 1 to 10.
12. Microcapsules according to claim 11 with diameter of between 0.2 to 8 µm.
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DE19545257A DE19545257A1 (en) | 1995-11-24 | 1995-11-24 | Process for the production of morphologically uniform microcapsules and microcapsules produced by this process |
DE19545257.7 | 1995-11-24 | ||
PCT/EP1996/004701 WO1997019676A1 (en) | 1995-11-24 | 1996-10-30 | Method of producing morphologically uniform microcapsules and microcapsules produced by this method |
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