MXPA96001756A - Urate-oxidase-containing stable liquid composition, and lyophilized composition for the preparationm thereof - Google Patents
Urate-oxidase-containing stable liquid composition, and lyophilized composition for the preparationm thereofInfo
- Publication number
- MXPA96001756A MXPA96001756A MXPA/A/1996/001756A MX9601756A MXPA96001756A MX PA96001756 A MXPA96001756 A MX PA96001756A MX 9601756 A MX9601756 A MX 9601756A MX PA96001756 A MXPA96001756 A MX PA96001756A
- Authority
- MX
- Mexico
- Prior art keywords
- composition according
- composition
- alanine
- poloxamer
- mannitol
- Prior art date
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 61
- 108010092464 Urate Oxidase Proteins 0.000 title claims abstract description 60
- 239000007788 liquid Substances 0.000 title claims abstract description 11
- 229940005267 urate oxidase Drugs 0.000 title claims description 59
- 229920001993 poloxamer 188 Polymers 0.000 claims abstract description 33
- 229940044519 Poloxamer 188 Drugs 0.000 claims abstract description 32
- 239000003125 aqueous solvent Substances 0.000 claims abstract description 11
- FBPFZTCFMRRESA-KVTDHHQDSA-N Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 29
- 229960003767 Alanine Drugs 0.000 claims description 28
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 28
- FBPFZTCFMRRESA-KAZBKCHUSA-N D-Mannitol Natural products OC[C@@H](O)[C@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KAZBKCHUSA-N 0.000 claims description 26
- 239000000594 mannitol Substances 0.000 claims description 26
- 235000010355 mannitol Nutrition 0.000 claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- 239000007924 injection Substances 0.000 claims description 16
- QNAYBMKLOCPYGJ-UWTATZPHSA-N L-Alanine Natural products C[C@@H](N)C(O)=O QNAYBMKLOCPYGJ-UWTATZPHSA-N 0.000 claims description 15
- 235000004279 alanine Nutrition 0.000 claims description 13
- DGLRDKLJZLEJCY-UHFFFAOYSA-L disodium;hydrogen phosphate;dodecahydrate Chemical compound O.O.O.O.O.O.O.O.O.O.O.O.[Na+].[Na+].OP([O-])([O-])=O DGLRDKLJZLEJCY-UHFFFAOYSA-L 0.000 claims description 12
- UIIMBOGNXHQVGW-UHFFFAOYSA-M buffer Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 7
- WVDDGKGOMKODPV-UHFFFAOYSA-N benzyl alcohol Chemical compound OCC1=CC=CC=C1 WVDDGKGOMKODPV-UHFFFAOYSA-N 0.000 claims description 6
- 239000012064 sodium phosphate buffer Substances 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 230000002335 preservative Effects 0.000 claims description 5
- 239000003755 preservative agent Substances 0.000 claims description 5
- LXCFILQKKLGQFO-UHFFFAOYSA-N Methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 claims description 4
- 238000011084 recovery Methods 0.000 claims description 4
- 229960000686 Benzalkonium Chloride Drugs 0.000 claims description 2
- 229960001950 Benzethonium Chloride Drugs 0.000 claims description 2
- UREZNYTWGJKWBI-UHFFFAOYSA-M Benzethonium chloride Chemical compound [Cl-].C1=CC(C(C)(C)CC(C)(C)C)=CC=C1OCCOCC[N+](C)(C)CC1=CC=CC=C1 UREZNYTWGJKWBI-UHFFFAOYSA-M 0.000 claims description 2
- RLSSMJSEOOYNOY-UHFFFAOYSA-N M-Cresol Chemical compound CC1=CC=CC(O)=C1 RLSSMJSEOOYNOY-UHFFFAOYSA-N 0.000 claims description 2
- QELSKZZBTMNZEB-UHFFFAOYSA-N Propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 claims description 2
- 235000019445 benzyl alcohol Nutrition 0.000 claims description 2
- 229960004217 benzyl alcohol Drugs 0.000 claims description 2
- 239000007972 injectable composition Substances 0.000 claims description 2
- 238000007918 intramuscular administration Methods 0.000 claims description 2
- 238000001990 intravenous administration Methods 0.000 claims description 2
- 229940100630 metacresol Drugs 0.000 claims description 2
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 claims description 2
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 claims description 2
- 229960002216 methylparaben Drugs 0.000 claims description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 2
- 229960003742 phenol Drugs 0.000 claims description 2
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 claims description 2
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 claims description 2
- 229960003415 propylparaben Drugs 0.000 claims description 2
- 238000007920 subcutaneous administration Methods 0.000 claims description 2
- 238000004090 dissolution Methods 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 36
- 239000002904 solvent Substances 0.000 description 11
- 230000002255 enzymatic Effects 0.000 description 10
- 238000005429 turbidity Methods 0.000 description 9
- 238000002360 preparation method Methods 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 239000002736 nonionic surfactant Substances 0.000 description 5
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 5
- 229920000053 polysorbate 80 Polymers 0.000 description 5
- 229940088598 Enzyme Drugs 0.000 description 4
- 230000000875 corresponding Effects 0.000 description 4
- 229940061607 Dibasic Sodium Phosphate Drugs 0.000 description 3
- 229940068968 Polysorbate 80 Drugs 0.000 description 3
- LEHOTFFKMJEONL-UHFFFAOYSA-N Trioxopurine Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 3
- 229940116269 Uric Acid Drugs 0.000 description 3
- 229920001577 copolymer Polymers 0.000 description 3
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000244 polyoxyethylene sorbitan monooleate Substances 0.000 description 3
- POJWUDADGALRAB-PVQJCKRUSA-N Allantoin Natural products NC(=O)N[C@@H]1NC(=O)NC1=O POJWUDADGALRAB-PVQJCKRUSA-N 0.000 description 2
- 241000228197 Aspergillus flavus Species 0.000 description 2
- 102000018997 Growth Hormone Human genes 0.000 description 2
- 108010051696 Growth Hormone Proteins 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 239000000122 growth hormone Substances 0.000 description 2
- 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 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- -1 polyoxyethylene Polymers 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 102000034363 protein filaments Human genes 0.000 description 2
- 108091005958 protein filaments Proteins 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 229960000458 Allantoin Drugs 0.000 description 1
- 229920002676 Complementary DNA Polymers 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N D-Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 102000002265 Human Growth Hormone Human genes 0.000 description 1
- 108010000521 Human Growth Hormone Proteins 0.000 description 1
- 239000000854 Human Growth Hormone Substances 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 210000003093 Intracellular Space Anatomy 0.000 description 1
- 210000003734 Kidney Anatomy 0.000 description 1
- GUBGYTABKSRVRQ-UUNJERMWSA-N Lactose Natural products O([C@@H]1[C@H](O)[C@H](O)[C@H](O)O[C@@H]1CO)[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1 GUBGYTABKSRVRQ-UUNJERMWSA-N 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 229920001451 Polypropylene glycol Polymers 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000721 bacterilogical Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229920001400 block copolymer Polymers 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000004027 cells Anatomy 0.000 description 1
- 230000000973 chemotherapeutic Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L disodium;2-[2-[carboxylatomethyl(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetate Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- MQRJBSHKWOFOGF-UHFFFAOYSA-L disodium;carbonate;hydrate Chemical compound O.[Na+].[Na+].[O-]C([O-])=O MQRJBSHKWOFOGF-UHFFFAOYSA-L 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 238000005755 formation reaction Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000002209 hydrophobic Effects 0.000 description 1
- 201000001431 hyperuricemia Diseases 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000000644 isotonic solution Substances 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 229960001855 mannitol Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000001264 neutralization Effects 0.000 description 1
- IAYPIBMASNFSPL-UHFFFAOYSA-N oxane Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 1
- 229920001983 poloxamer Polymers 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- SDRZXZKXVBHREH-UHFFFAOYSA-J potassium;dihydrogen phosphate;phosphate Chemical compound [K+].OP(O)([O-])=O.[O-]P([O-])([O-])=O SDRZXZKXVBHREH-UHFFFAOYSA-J 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- GOOHAUXETOMSMM-UHFFFAOYSA-N propylene oxide Chemical compound CC1CO1 GOOHAUXETOMSMM-UHFFFAOYSA-N 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 230000003381 solubilizing Effects 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000001225 therapeutic Effects 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
Abstract
The present invention refers to a physically stable, pharmaceutically acceptable liquid composition comprising urate-oxidases, and from 0.1 mg/ml to 10 mg/ml of Poloxamer 188, in a buffered aqueous medium. This composition can be obtained by dissolving a lyophilized compound in an aqueous solvent.
Description
REF: 22475
1 STABLE LIQUID COMPOSITION CONTAINING URATO-OXIDASE, AND LYOPHILIZED COMPOSITION FOR PREPARATION
FIELD AND BACKGROUND OF THE INVENTION
The present invention relates to a liquid pharmaceutically acceptable formulation, which contains urate oxidase, which is in the form of a solution stable at 25 ° C and clear after agitation. This solution can also be obtained by dissolving a lyophilisate containing the urate oxidase in a solvent. 5 Urate-oxidase (urate-oxygen-oxidoreductase
EC 1.7.3.3. Uox), a protein enzyme obtained from Aspergillus flavus that oxidizes uric acid to alantoin, is used for the prevention or treatment of hyperuricaemia at the same time as chemotherapies, Q since allantoin is ten times more soluble that uric acid, and easily eliminated by the kidneys (Laboureur P. et al, Bull. Soc. Chim. Biol., 1968, 50, 811-825; Kissel P. et al., The Lancet, 1975, J25, 229). -e Urate-oxidase is a tetrameric enzyme composed of four identical units with a molecular weight of 34,152. Each monomeric unit, formed of a single polypeptide chain of 301 amino acids, is acetylated at the N-terminus, and has no disulfide bridges. The optimum pH of stability of the enzymatic activity of the urate oxidase in solution is pH = 8 (Bayol, A. and collaborators, accepted for publication Biophys, Chem., 1995 (54), 229-35). The cDNA encoding this protein has been cloned and expressed in E. coli (Legoux, R., et al J. Biol. Chem., 1992, 267, (12), 8565-8570); in A. flavus (Chevalet, L. et al, Curr. Genet, 1992, 21, 447-453); and in S. cerevisiae (Leplatois, P. et al., Gene., 1992, 122, 139-145). The best yields obtained from S. cerevisiae have favored the selection of this yeast for the production of recombinant urate oxidase (re-Uox). The recombinant enzyme accumulated in the intracellular space under a soluble and active form is extracted and then purified until a pharmaceutical quality is obtained. In order to obtain a formulation administrable to man, the urate oxidase must be prepared in the form of a pharmaceutical composition. Such compositions must retain the enzymatic activity - of the urate oxidase over time. A lyophilized form is generally used to preserve the biochemical integrity and biological activity of the enzymes under very varied preservation conditions. It is well known that lyophilized preparations retain these properties better than the corresponding liquid preparations. The lyophilized preparations must be diluted in a pharmaceutically acceptable solvent, before administration to man by the parenteral route. A liquid composition of the enzyme will however be preferable since this will be more rapidly administrable, with the condition nonetheless of being physically and chemically stable. The known pharmaceutical formulation of urate oxidase (Uricozyme) is a lyophilized injectable preparation that needs a reconstitution solvent. The composition of the freeze-dried form of Uricozyme is as follows: Urato-oxidase 1000 units
Aza-8-dioxo-2,6-purine monohydrate Sodium carbonate neutral anhydrous Tetracemate disodium dihydrate 0.37 mg
Lactose The composition of the reconstitution solvent is as follows: Potassium monoacid phosphate Potassium dihydrogen phosphate Glucose Water for injections c. s. s. 1 mi
(VIDAL Paris Dictionary 61st edition P. 1552, 1985).
The solution of urate oxidase obtained after the solution of the lyophilisate in the reconstitution solvent will increase its turbidity with the storage time at 25 ° C, until the appearance of a filamentous protein precipitate. The formation of this protein precipitate in solution is accelerated with time and the force of agitation of the solution. The solution obtained is physically unstable. The solubilizing properties of non-ionic surfactant molecules are well known to the person skilled in the art., but the interactions between proteins and non-ionic surfactants are more specific and described in the literature. European Patent EP 211 601 describes the use of certain block copolymers, which are non-ionic surfactant molecules, in the stabilization of injectable formulations of growth hormone. The growth hormone is stabilized in a gelled matrix formed by a sequenced copolymer containing polyoxyethylenated and polyoxypropylenated units of average molecular weight between 1, 100 and 40,000. The U.S. No. 4,783,441 describes a method of preventing the denaturation of proteins such as insulin in aqueous solution, by adding an amount that can reach up to 500 ppm of surfactant molecules composed of slightly hydrophilic and slightly hydrophobic alternating groups, in pH solutions. comprised between 6.8 and 8. US Pat. No. 5,096,885 describes the composition of a lyophilized solution containing human growth hormone, glycine, mannitol, a non-ionic surfactant and a buffer.
DESCRIPTION OF THE INVENTION
The objective of the present invention is a pharmaceutically acceptable liquid composition of a urate oxidase solution containing between 0.1 mg / ml and 10 mg / ml of a sequenced copolymer of ethylene oxide and propylene oxide, Poloxamer 188, in watery medium cushioned. This composition is clear and stable physically and chemically after vigorous agitation or after storage, for at least 48 hours and up to one year at 25 ° C. The advantages offered by this form are a greater simplicity of use in clinic, together with the possibility of being administered directly and of being stored under its directly administrable form, during longer periods of time. Another advantage lies in the absence of particular instructions for maintenance precautions, usually necessary to avoid the precipitation of protein filaments in the solution to be injected, since the composition according to the invention has the property of preventing the precipitation of protein filaments in the sine of the solution. The aqueous liquid composition of the invention is clear, stable, sterile and pharmaceutically acceptable, and can be injected into man or an animal by the subcutaneous, intravenous or intramuscular route. In the present invention, the term "urate oxidase" is understood to mean the protein obtained by fermentation of a natural or mutated strain by genetic engineering. The enzyme is therefore produced by the extraction of the natural source, or from cell cultures. The urate oxidase used in the formulations of the present invention can be obtained, for example, according to US 3,810,820, DD 284 689, DD 296 804, DD 300 781, DE 2 164 018, DE 1 517 742, FR 2 664 286, GB 2 221 910, JP 76-007749, JP-75-030137, JP-84-023987, JP73 018473, JP-47 029575, SU 565 935, US 4,062,731, EP 545 688 or according to EP 435 776. The term "pharmaceutically effective amount" refers to any amount of urate oxidase that produces a therapeutic effect. The solutions corresponding to the invention contain a pharmaceutically effective amount of urate oxidase. Preferably these solutions contain between 0.1 mg / ml and 50 mg / ml of urate oxidase according to the desired dose. The concentration range of the urate oxidase is not critical to the invention and may vary according to the preparations. The solutions according to the invention contain a nonionic surfactant of the type copolymer sequence of polyoxyethylene and polyoxypropylene, called Poloxamer 188 and corresponding to the formula H0 (CH2CH20) 75 (CH (CH3) CH20) (CH2CH20) 75 H. product is marketed by the company ICI under the name Synperonic F 68 and by the company B.A.S.F. under the name of Pluronic F 68. The necessary quantities of Poloxamer 188 to obtain physically stable solutions of urate oxidase are between 0.1 mg / ml and 10 mg / ml, preferably between 0.5 mg / ml and 5 mg / ml. The pH of the solution is preferably between 7.5 and 8.5. When the pH is equal to 8, the urate oxidase is particularly stable chemically in solution. Generally, it is preferred to use a sodium phosphate buffer to buffer the medium. The concentration of the buffer is advantageously from 5 mM to 100 mM. The buffer used is preferably a sodium phosphate buffer of pH = 8, at a concentration between 5 mM and 100 mM. The liquid composition according to the invention is preferably isotonic and advantageously comprises the excipients necessary for obtaining an isotonic solution, such as mannitol or alanine. The solution may contain in particular mannitol, preferably at a concentration between 1 mg / ml and 50 mg / ml.
- -
The solution may contain, in particular, alanine, preferably at a concentration between 1 mg / ml and 50 mg / ml. The solution preferably contains mannitol and alanine in any proportion that conserves the isoto-nicity of the solution, preferably from 1 to 50 mg / ml of mannitol and from 1 to 50 mg / ml of alanine. Advantageously, the solution contains approximately 10.6 mg / ml of mannitol and approximately 15.9 mg / ml of alanine. The liquid composition according to the invention advantageously comprises the preservatives necessary for the bacteriological preservation of the solution, mainly such as benzyl alcohol, phenol, meta-cresol, methyl-paraben, propyl-paraben, benzalkonium chloride or benzethonium chloride. The solution can be obtained directly by dissolving the constituents in water; or after reconstitution of a lyophilisate containing urate oxidase, by means of an aqueous solvent containing Poloxamer 188; or after reconstitution of a lyophilisate containing urate oxidase and Poloxamer 188, by means of an aqueous solvent. In this regard, the invention also aims at a lyophilized composition for dissolving - - in an aqueous solvent, and containing urate oxidase and Poloxamer 188, the weight ratio of Poloxamer 188 to urate oxidase being 0.01 to 50. The lyophilized composition may advantageously comprise any other buffer, as indicated above for the solution. It can also advantageously comprise the excipients which ensure the isotonicity of. the aqueous solution obtained by dissolving the lyophilisate in an aqueous solvent, such as, for example, alanine or mannitol, in variable amounts depending on the volume of reconstitution solvent to be used. The following examples illustrate the invention, without limiting it.
EXAMPLE 1
Urate-oxidase solution prepared by dissolving the constituents in water
Urate-oxidase 1.5 mg
Manitol 10.6 mg
L-alanine 15.9 mg
Dibasic sodium phosphate dodecahydrate 14.32 mg
Poloxamer 188 1 mg - -
Water for injections c.s. 1 mi
EXAMPLE 2
Urate-oxidase solution prepared from a lyophilisate of urate-oxidase reconstituted with an aqueous recovery solvent containing Poloxamer 188.
Composition of lyophilisate:
Urate-oxidase 1.5 mg
Manitol 10.6 mg
L-alanine 15.9 mg Dibasic sodium phosphate dodecahydrate 14.32 mg
Composition of the recovery solvent:
Poloxamer 188 1 mg Water for injections c.s. 1 mi
EXAMPLE 3
Urate-oxidase solution prepared from a lyophilisate of urate oxidase containing the - - Poloxamer 188 reconstituted with water.
Composition of lyophilisate:
Urate-oxidase 1.5 mg
Manitol 10.6 mg
L-alanine 15.9 mg
Dibasic sodium phosphate dodecahydrate 14.32 mg
Poloxamer 188 1 mg
Composition of the recovery solvent:
Water for injections c.s. 1 mi
The advantages of the compositions of the invention are now to be evidenced by comparing the properties of different compositions based on urate oxidase, summarized in Table 1 and in the appended figures.
PREPARATIONS OF THE COMPOSITIONS
The urate oxidase solutions according to the invention can be obtained in different ways, for example: by mixing a concentrated aqueous solution of urate oxidase containing a sodium phosphate buffer at pH. = 8, with an aqueous solution containing Poloxamer 188, and optionally with excipients that ensure the isotonicity of the mixed solution, and optionally preservatives. by dissolving a lyophilisate containing urate oxidase, a sodium phosphate buffer at pH = 8, ballast excipients that ensure isotonicity, such as alanine and mannitol; in an aqueous solvent containing Poloxamer 188 and possibly preservatives. by dissolving a lyophilisate containing urate oxidase, a sodium phosphate buffer at pH = 8, ballast excipients such as alanine and mannitol, and Poloxamer 188; in an aqueous solvent that eventually contains preservatives.
The solutions obtained by these three procedures can be sterile filtered. These retain their physical stability as well as the chemical stability and enzymatic activity of the urate oxidase for 1 month at 25 ° C. Different analytical methods have been used to measure different parameters, such as turbidity or the dosage of enzymatic activity
1. Turbidity measurement:
The turbidity of the urate oxidase solutions is determined by a Hach Ratio turbidimeter. The turbidity results are expressed in Nephelometric Turbidity Units (NTU) defined by: Standard methods for the examination of water and wastewater from the American Association for Public Health. The turbidity measurement indicates the degree of aggregation of the urate oxidase in solution.
2. Dosage of the enzymatic activity of urate oxidase:
The enzymatic activity of urate oxidase is determined by spectrophotometry in a thermostatic cell at 30 ° C, following the disappearance of uric acid at 292 nm (Legoux, R. et al., J. Biol. Chem., 1992, 267 (12 8565-8570).
The stability tests of the reconstituted solute after Example 2 were carried out at 25 ° C. Table I below shows the stability of the reconstituted solutes by dissolving with water and with solvents respectively containing 1 mg of Polysorbate 80 (Tween 80) or 1 mg of Poloxamer 188 according to the invention, with a sufficient amount of water to produce a volume of solute of 1 ml.
Table I: Stability of the reconstituted solute at 25 ° C
Time 0 24 hours 48 hours UAE / ml UAE / ml UAE / ml Solvent Appearance pH activity Appearance pH activity Appearance pH Enzymatic enzymatic enzymatic activity
Water > IV 8.01 26.1 > IV 8.01 26.5 > IV 8.01 27.9
Tween 80 0.1% II 8.0 26.0 II to III 8.0. 28.1 III to IV 8.0 29.1 I
Poloxamer 188 II 8.0 24.7 II 8.0 28.9 II 8.0 28.5 to 0.1%
UAE: Enzymatic activity unit
The appearance (opalescence) is determined according to the method of European Pharmacopoeia (II) V.6 by comparing the sample to be analyzed with a control suspension.
DESCRIPTION OF THE FIGURES
FIGURE NQ 1 indicates the evolution during 24 hours of turbidity in NTU, after the stirring during 1 minute in a vortex apparatus, of five solutions of urate oxidase corresponding or not to the invention, depending on their proportions of surfactants non-ionic
Composition NQ 1: 15 mg of urate oxidase,
106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate and 10 ml of water for injections.
Composition NQ 2: 15 mg of urate oxidase, 106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate, 10 mg of Polysorbate 80 and 10 ml of water for injections.
Composition Ns 3 (according to the invention): 15 mg of urate oxidase, 106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate, 10 mg of Poloxamer 188 and 10 ml of water for injections .
Composition Ns 4: 15 mg of urate oxidase, 106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate, 100 mg of Polysorbate 80 and 10 ml of water for injections.
Na 5 composition (according to the invention): 15 mg of urate oxidase, 106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate, 100 mg of Poloxamer 188 and 10 ml of water for injections .
FIGURE Na 2 indicates the evolution during 24 hours of the turbidity in NTU, after the stirring during 1 minute in a vortex apparatus, of six solutions of urate oxidase containing Poloxaraer 188 according to the invention, depending on its proportion of Poloxamer 188.
Composition NQ 5: 15 mg of urate oxidase, 106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate, 100 mg of Poloxamer 188 and 10 ml of water for injections.
Composition Ns 3: 15 mg of urate oxidase, 106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate, 10 mg of Poloxamer
188 and 10 ml of water for injections.
Composition Nd 6: 15 mg of urate oxidase, 106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate, 7.5 mg of Poloxamer
188 and 10 ml of water for injections.
Composition NQ 7: 15 mg of urate oxidase, 106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate, 5 mg of Poloxamer
188 and 10 ml of water for injections.
Composition Ns 8: 15 mg 'of urate oxidase, 106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate, 2.5 mg of Poloxamer 188 and 10 ml of water for injections.
Composition NQ 9: 15 mg of urate oxidase, 106 mg of mannitol, 159 mg of L-alanine, 143.2 mg of disodium phosphate dodecahydrate, 1 mg of Poloxamer 188 and 10 ml of water for injections.
It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention. Having described the invention as above, property is claimed as contained in the following:
Claims (20)
1. A pharmaceutically acceptable, physically stable liquid composition, characterized in that it contains urate oxidase and 0.1 mg / ml to 10 mg / ml of Poloxamer 188, in a buffered aqueous medium.
2. The composition according to claim 1, characterized in that it contains between 0.5 mg / ml and 5 mg / ml Poloxamer 188.
3. The composition according to claim 1, characterized in that it also contains alanine.
4. The composition according to claim 3, characterized in that in this the amount of alanine is between 1 mg / ml and 50 mg / ml.
5. The composition according to claim 1, characterized in that it also contains mannitol.
The composition according to claim 5, characterized in that therein the amount of mannitol is between 1 mg / ml and 50 mg / ml.
7. The composition according to claim 1, characterized in that it also contains alanine and mannitol.
8. The composition according to claim 7, characterized in that it contains from 1 to 50 mg / ml of alanine and from 1 to 50 mg / ml of mannitol.
9. The composition according to claim 8, characterized in that it contains 15.9 mg / ml of alanine and 10.6 mg / ml of mannitol.
10. The composition according to claim 1, characterized in that it is isotonic.
11. The composition according to claim 1, characterized in that it contains a sodium phosphate buffer.
12 The composition according to claim 1, characterized in that in this the concentration of the buffer is between 5 mM and 100 mM.
13. The composition according to claim 1, characterized in that the pH is between 7.5 and 8.5.
14. The composition according to claim 1, characterized in that it contains one or more preservatives chosen from phenol, benzyl alcohol, meta-cresol, methyl-paraben, propyl-paraben, benzalkonium chloride or benzethonium chloride.
15. The composition according to claim 1, characterized in that it is obtained by dissolving a lyophilisate in an aqueous solvent.
16. The composition according to claim 1, characterized in that it is sterile and injectable to man or animal via the subcutaneous, intravenous or intramuscular route.
17. The injectable composition in accordance - - with claim 15 and claim 16, characterized in that in this the composition of the lyophilizate is: Urate-oxidase 1.5 mg Mannitol 10.6 mg L-alanine 15.9 mg Disodium phosphate dodecahydrate 14.32 mg and the composition of the recovery aqueous solvent is: Poloxamer 188 1 mg Water for injections c.s. 1 mi
18. The lyophilized composition for dissolution in an aqueous solvent, characterized in that it contains urate oxidase and Poloxamer 188, the weight ratio of Poloxamer 188 to urate oxidase being 0.01 to 50.
19. The lyophilized composition according to claim 18, characterized in that it also contains a buffer.
20. The lyophilized composition according to claim 18, characterized in that it also contains excipients that ensure the isotonicity of the aqueous solution obtained by dissolving the lyophilizate in an aqueous solvent.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR9505606 | 1995-05-11 | ||
| FR9505606A FR2733914B1 (en) | 1995-05-11 | 1995-05-11 | STABLE LIQUID COMPOSITION CONTAINING URATE OXYDASE AND LYOPHILIZED COMPOSITION FOR ITS PREPARATION |
| US08/644,163 US5811096A (en) | 1995-05-11 | 1996-05-10 | Stable liquid composition containing urate oxidase and lyophilized composition for its preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| MX9601756A MX9601756A (en) | 1997-07-31 |
| MXPA96001756A true MXPA96001756A (en) | 1997-12-01 |
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