WO2007033427A1 - Stabilisation of peptides with basic amino acids - Google Patents
Stabilisation of peptides with basic amino acids Download PDFInfo
- Publication number
- WO2007033427A1 WO2007033427A1 PCT/AU2006/001391 AU2006001391W WO2007033427A1 WO 2007033427 A1 WO2007033427 A1 WO 2007033427A1 AU 2006001391 W AU2006001391 W AU 2006001391W WO 2007033427 A1 WO2007033427 A1 WO 2007033427A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- peptide
- amino acid
- basic amino
- formulation according
- formulation
- Prior art date
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 97
- 150000001413 amino acids Chemical class 0.000 title claims abstract description 64
- 102000004196 processed proteins & peptides Human genes 0.000 title description 18
- 230000006641 stabilisation Effects 0.000 title description 2
- 239000000203 mixture Substances 0.000 claims abstract description 124
- 238000009472 formulation Methods 0.000 claims abstract description 104
- 239000012458 free base Substances 0.000 claims abstract description 28
- 238000000034 method Methods 0.000 claims abstract description 24
- 238000005903 acid hydrolysis reaction Methods 0.000 claims abstract description 20
- 239000004480 active ingredient Substances 0.000 claims abstract description 13
- 230000015556 catabolic process Effects 0.000 claims abstract description 9
- 238000006731 degradation reaction Methods 0.000 claims abstract description 9
- 235000001014 amino acid Nutrition 0.000 claims description 58
- 239000004475 Arginine Substances 0.000 claims description 44
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 44
- 108010070305 AOD 9604 Proteins 0.000 claims description 37
- GVIYUKXRXPXMQM-BPXGDYAESA-N 221231-10-3 Chemical group C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]1C(N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CSSC1)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C(C)C)=O)C1=CC=C(O)C=C1 GVIYUKXRXPXMQM-BPXGDYAESA-N 0.000 claims description 36
- 239000002775 capsule Substances 0.000 claims description 17
- 150000003839 salts Chemical group 0.000 claims description 9
- 239000007787 solid Substances 0.000 claims description 6
- -1 Ornthine Chemical compound 0.000 claims description 5
- 150000002632 lipids Chemical class 0.000 claims description 5
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims description 3
- RHGKLRLOHDJJDR-BYPYZUCNSA-N L-citrulline Chemical compound NC(=O)NCCC[C@H]([NH3+])C([O-])=O RHGKLRLOHDJJDR-BYPYZUCNSA-N 0.000 claims description 3
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 claims description 3
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 claims description 3
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 3
- 239000004472 Lysine Substances 0.000 claims description 3
- RHGKLRLOHDJJDR-UHFFFAOYSA-N Ndelta-carbamoyl-DL-ornithine Natural products OC(=O)C(N)CCCNC(N)=O RHGKLRLOHDJJDR-UHFFFAOYSA-N 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 3
- 229960002173 citrulline Drugs 0.000 claims description 3
- 235000013477 citrulline Nutrition 0.000 claims description 3
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 3
- 108010000521 Human Growth Hormone Proteins 0.000 claims description 2
- 102000002265 Human Growth Hormone Human genes 0.000 claims description 2
- 239000000854 Human Growth Hormone Substances 0.000 claims description 2
- AHLPHDHHMVZTML-BYPYZUCNSA-N L-Ornithine Chemical compound NCCC[C@H](N)C(O)=O AHLPHDHHMVZTML-BYPYZUCNSA-N 0.000 claims description 2
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 claims description 2
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 claims description 2
- 210000001789 adipocyte Anatomy 0.000 claims description 2
- 230000004132 lipogenesis Effects 0.000 claims description 2
- 230000004130 lipolysis Effects 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 229960003104 ornithine Drugs 0.000 claims description 2
- 229960003121 arginine Drugs 0.000 claims 1
- 229960002885 histidine Drugs 0.000 claims 1
- 229960003646 lysine Drugs 0.000 claims 1
- 235000009697 arginine Nutrition 0.000 description 41
- 229940024606 amino acid Drugs 0.000 description 40
- 239000000243 solution Substances 0.000 description 21
- 238000006243 chemical reaction Methods 0.000 description 20
- 239000003826 tablet Substances 0.000 description 13
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 12
- 239000011347 resin Substances 0.000 description 12
- 229920005989 resin Polymers 0.000 description 12
- 239000007963 capsule composition Substances 0.000 description 10
- 150000001875 compounds Chemical class 0.000 description 10
- 239000000843 powder Substances 0.000 description 10
- 238000003860 storage Methods 0.000 description 10
- 239000007916 tablet composition Substances 0.000 description 9
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 7
- 239000000546 pharmaceutical excipient Substances 0.000 description 7
- 210000002345 respiratory system Anatomy 0.000 description 7
- DTQVDTLACAAQTR-UHFFFAOYSA-N trifluoroacetic acid Substances OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 238000003556 assay Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 4
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 239000011777 magnesium Substances 0.000 description 4
- 239000008194 pharmaceutical composition Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 3
- 229930064664 L-arginine Natural products 0.000 description 3
- 235000014852 L-arginine Nutrition 0.000 description 3
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 3
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 235000010443 alginic acid Nutrition 0.000 description 3
- 229920000615 alginic acid Polymers 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 238000000354 decomposition reaction Methods 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000003937 drug carrier Substances 0.000 description 3
- 229940112141 dry powder inhaler Drugs 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 239000000314 lubricant Substances 0.000 description 3
- 229910052749 magnesium Inorganic materials 0.000 description 3
- 235000001055 magnesium Nutrition 0.000 description 3
- 239000008108 microcrystalline cellulose Substances 0.000 description 3
- 229940016286 microcrystalline cellulose Drugs 0.000 description 3
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 229920001223 polyethylene glycol Polymers 0.000 description 3
- 229920001592 potato starch Polymers 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 238000004007 reversed phase HPLC Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- CSMYOORPUGPKAP-IBGZPJMESA-N (2r)-3-(acetamidomethylsulfanyl)-2-(9h-fluoren-9-ylmethoxycarbonylamino)propanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CSCNC(=O)C)C(O)=O)C3=CC=CC=C3C2=C1 CSMYOORPUGPKAP-IBGZPJMESA-N 0.000 description 2
- LNAZSHAWQACDHT-XIYTZBAFSA-N (2r,3r,4s,5r,6s)-4,5-dimethoxy-2-(methoxymethyl)-3-[(2s,3r,4s,5r,6r)-3,4,5-trimethoxy-6-(methoxymethyl)oxan-2-yl]oxy-6-[(2r,3r,4s,5r,6r)-4,5,6-trimethoxy-2-(methoxymethyl)oxan-3-yl]oxyoxane Chemical compound CO[C@@H]1[C@@H](OC)[C@H](OC)[C@@H](COC)O[C@H]1O[C@H]1[C@H](OC)[C@@H](OC)[C@H](O[C@H]2[C@@H]([C@@H](OC)[C@H](OC)O[C@@H]2COC)OC)O[C@@H]1COC LNAZSHAWQACDHT-XIYTZBAFSA-N 0.000 description 2
- SWZCTMTWRHEBIN-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-(4-hydroxyphenyl)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C1=CC=C(O)C=C1 SWZCTMTWRHEBIN-QFIPXVFZSA-N 0.000 description 2
- QTWZCODKTSUZJN-LJAQVGFWSA-N (2s)-5-[[amino-[(2,2,5,7,8-pentamethyl-3,4-dihydrochromen-6-yl)sulfonylamino]methylidene]amino]-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N[C@H](C(O)=O)CCCN=C(N)NS(=O)(=O)C(C(C)=C1C)=C(C)C2=C1OC(C)(C)CC2 QTWZCODKTSUZJN-LJAQVGFWSA-N 0.000 description 2
- VBICKXHEKHSIBG-UHFFFAOYSA-N 1-monostearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(O)CO VBICKXHEKHSIBG-UHFFFAOYSA-N 0.000 description 2
- IZHVBANLECCAGF-UHFFFAOYSA-N 2-hydroxy-3-(octadecanoyloxy)propyl octadecanoate Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(O)COC(=O)CCCCCCCCCCCCCCCCC IZHVBANLECCAGF-UHFFFAOYSA-N 0.000 description 2
- QCQCHGYLTSGIGX-GHXANHINSA-N 4-[[(3ar,5ar,5br,7ar,9s,11ar,11br,13as)-5a,5b,8,8,11a-pentamethyl-3a-[(5-methylpyridine-3-carbonyl)amino]-2-oxo-1-propan-2-yl-4,5,6,7,7a,9,10,11,11b,12,13,13a-dodecahydro-3h-cyclopenta[a]chrysen-9-yl]oxy]-2,2-dimethyl-4-oxobutanoic acid Chemical compound N([C@@]12CC[C@@]3(C)[C@]4(C)CC[C@H]5C(C)(C)[C@@H](OC(=O)CC(C)(C)C(O)=O)CC[C@]5(C)[C@H]4CC[C@@H]3C1=C(C(C2)=O)C(C)C)C(=O)C1=CN=CC(C)=C1 QCQCHGYLTSGIGX-GHXANHINSA-N 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 2
- AAAFQLPJNOITCL-SFHVURJKSA-N 9h-fluoren-9-ylmethyl n-[(2s)-1-oxo-3-phenylpropan-2-yl]carbamate Chemical compound C([C@@H](C=O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C1=CC=CC=C1 AAAFQLPJNOITCL-SFHVURJKSA-N 0.000 description 2
- 239000005995 Aluminium silicate Substances 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 2
- 239000001856 Ethyl cellulose Substances 0.000 description 2
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound 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
- 239000003875 Wang resin Substances 0.000 description 2
- NERFNHBZJXXFGY-UHFFFAOYSA-N [4-[(4-methylphenyl)methoxy]phenyl]methanol Chemical compound C1=CC(C)=CC=C1COC1=CC=C(CO)C=C1 NERFNHBZJXXFGY-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 2
- 239000000783 alginic acid Substances 0.000 description 2
- 229960001126 alginic acid Drugs 0.000 description 2
- 150000004781 alginic acids Chemical class 0.000 description 2
- 235000012211 aluminium silicate Nutrition 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 239000001506 calcium phosphate Substances 0.000 description 2
- 229910000389 calcium phosphate Inorganic materials 0.000 description 2
- 235000011010 calcium phosphates Nutrition 0.000 description 2
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 2
- 235000010980 cellulose Nutrition 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 238000002144 chemical decomposition reaction Methods 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 230000006240 deamidation Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 230000007850 degeneration Effects 0.000 description 2
- 230000000593 degrading effect Effects 0.000 description 2
- UNXNGGMLCSMSLH-UHFFFAOYSA-N dihydrogen phosphate;triethylazanium Chemical compound OP(O)(O)=O.CCN(CC)CC UNXNGGMLCSMSLH-UHFFFAOYSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 238000009505 enteric coating Methods 0.000 description 2
- 239000002702 enteric coating Substances 0.000 description 2
- 235000019325 ethyl cellulose Nutrition 0.000 description 2
- 229920001249 ethyl cellulose Polymers 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229940014259 gelatin Drugs 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 2
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 2
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 2
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 229910052740 iodine Inorganic materials 0.000 description 2
- 239000011630 iodine Substances 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 229920000609 methyl cellulose Polymers 0.000 description 2
- 235000010981 methylcellulose Nutrition 0.000 description 2
- 239000001923 methylcellulose Substances 0.000 description 2
- 239000001788 mono and diglycerides of fatty acids Substances 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000002516 radical scavenger Substances 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 239000007909 solid dosage form Substances 0.000 description 2
- 235000010356 sorbitol Nutrition 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- 238000001694 spray drying Methods 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 2
- SJVFAHZPLIXNDH-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-phenylpropanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C1=CC=CC=C1 SJVFAHZPLIXNDH-QFIPXVFZSA-N 0.000 description 1
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 1
- NDKDFTQNXLHCGO-UHFFFAOYSA-N 2-(9h-fluoren-9-ylmethoxycarbonylamino)acetic acid Chemical compound C1=CC=C2C(COC(=O)NCC(=O)O)C3=CC=CC=C3C2=C1 NDKDFTQNXLHCGO-UHFFFAOYSA-N 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- 229940117976 5-hydroxylysine Drugs 0.000 description 1
- TYMIFYJJOPYXBG-UHFFFAOYSA-N 9h-fluoren-9-ylmethyl piperidine-1-carboxylate Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N1CCCCC1 TYMIFYJJOPYXBG-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-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
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000282817 Bovidae Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 229920000623 Cellulose acetate phthalate Polymers 0.000 description 1
- 229920002785 Croscarmellose sodium Polymers 0.000 description 1
- 108010036941 Cyclosporins Proteins 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 150000008574 D-amino acids Chemical class 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical class Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 1
- 229920001479 Hydroxyethyl methyl cellulose Polymers 0.000 description 1
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- QUOGESRFPZDMMT-UHFFFAOYSA-N L-Homoarginine Natural products OC(=O)C(N)CCCCNC(N)=N QUOGESRFPZDMMT-UHFFFAOYSA-N 0.000 description 1
- 150000008575 L-amino acids Chemical class 0.000 description 1
- QUOGESRFPZDMMT-YFKPBYRVSA-N L-homoarginine Chemical compound OC(=O)[C@@H](N)CCCCNC(N)=N QUOGESRFPZDMMT-YFKPBYRVSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical group OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 1
- 240000007472 Leucaena leucocephala Species 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 229920002562 Polyethylene Glycol 3350 Polymers 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 229920001800 Shellac Polymers 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- CTQNLXWEQXSGSA-YBHXOQPNSA-N [(4s)-4-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2r)-2-amino-3-sulfanylidenepropanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-3-hydroxypropanoyl]amino]-3-methylbutanoyl]amino]-5-[[2-[[(2s)-1-[(2s)-2-[[2-[[(2r)-2-amino-3-sulfanylpropanoyl]amino]acetyl]amin Chemical compound NC(N)=NCCC[C@H](NC(=O)[C@@H](N)CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)OC(=O)CC[C@H](NC(=O)[C@@H](NC(=O)[C@H](CO)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)C=S)C(C)C)C(=O)NCC(=O)N[C@@H](CO)C(=O)OC(=O)[C@@H](NC(=O)CNC(=O)[C@@H](N)CS)CC1=CC=CC=C1 CTQNLXWEQXSGSA-YBHXOQPNSA-N 0.000 description 1
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 125000003342 alkenyl group Chemical group 0.000 description 1
- 125000005210 alkyl ammonium group Chemical group 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 125000000304 alkynyl group Chemical group 0.000 description 1
- PZZYQPZGQPZBDN-UHFFFAOYSA-N aluminium silicate Chemical compound O=[Al]O[Si](=O)O[Al]=O PZZYQPZGQPZBDN-UHFFFAOYSA-N 0.000 description 1
- 229910000323 aluminium silicate Inorganic materials 0.000 description 1
- 125000003277 amino group Chemical class 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000000181 anti-adherent effect Effects 0.000 description 1
- 230000003466 anti-cipated effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 125000003710 aryl alkyl group Chemical group 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 125000003289 ascorbyl group Chemical group [H]O[C@@]([H])(C([H])([H])O*)[C@@]1([H])OC(=O)C(O*)=C1O* 0.000 description 1
- CREXVNNSNOKDHW-UHFFFAOYSA-N azaniumylideneazanide Chemical group N[N] CREXVNNSNOKDHW-UHFFFAOYSA-N 0.000 description 1
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid group Chemical group C(C1=CC=CC=C1)(=O)O WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 239000004067 bulking agent Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 235000010216 calcium carbonate Nutrition 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 229920003123 carboxymethyl cellulose sodium Polymers 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 229940105329 carboxymethylcellulose Drugs 0.000 description 1
- 229940063834 carboxymethylcellulose sodium Drugs 0.000 description 1
- 230000006652 catabolic pathway Effects 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 229940081734 cellulose acetate phthalate Drugs 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000001055 chewing effect Effects 0.000 description 1
- 108050003126 conotoxin Proteins 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 229960001681 croscarmellose sodium Drugs 0.000 description 1
- 235000010947 crosslinked sodium carboxy methyl cellulose Nutrition 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 229930182912 cyclosporin Natural products 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- YSMODUONRAFBET-UHFFFAOYSA-N delta-DL-hydroxylysine Natural products NCC(O)CCC(N)C(O)=O YSMODUONRAFBET-UHFFFAOYSA-N 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 238000007907 direct compression Methods 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000000890 drug combination Substances 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- YSMODUONRAFBET-UHNVWZDZSA-N erythro-5-hydroxy-L-lysine Chemical compound NC[C@H](O)CC[C@H](N)C(O)=O YSMODUONRAFBET-UHNVWZDZSA-N 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 238000009501 film coating Methods 0.000 description 1
- 239000007888 film coating Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 230000009969 flowable effect Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 229910021485 fumed silica Inorganic materials 0.000 description 1
- 239000007903 gelatin capsule Substances 0.000 description 1
- 235000001727 glucose Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 125000000404 glutamine group Chemical group N[C@@H](CCC(N)=O)C(=O)* 0.000 description 1
- 229940074045 glyceryl distearate Drugs 0.000 description 1
- 229940075507 glyceryl monostearate Drugs 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 229940093915 gynecological organic acid Drugs 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000003906 humectant Substances 0.000 description 1
- 239000008172 hydrogenated vegetable oil Substances 0.000 description 1
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 1
- UWYVPFMHMJIBHE-OWOJBTEDSA-N hydroxymaleic acid group Chemical group O/C(/C(=O)O)=C/C(=O)O UWYVPFMHMJIBHE-OWOJBTEDSA-N 0.000 description 1
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 1
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 1
- 229920003132 hydroxypropyl methylcellulose phthalate Polymers 0.000 description 1
- 229940031704 hydroxypropyl methylcellulose phthalate Drugs 0.000 description 1
- 229920000639 hydroxypropylmethylcellulose acetate succinate Polymers 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 239000003701 inert diluent Substances 0.000 description 1
- 238000013101 initial test Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229920003145 methacrylic acid copolymer Polymers 0.000 description 1
- 229940117841 methacrylic acid copolymer Drugs 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- UBLQIESZTDNNAO-UHFFFAOYSA-N n,n-diethylethanamine;phosphoric acid Chemical compound [O-]P([O-])([O-])=O.CC[NH+](CC)CC.CC[NH+](CC)CC.CC[NH+](CC)CC UBLQIESZTDNNAO-UHFFFAOYSA-N 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid group Chemical group C(CCCCCCC\C=C/CCCCCCCC)(=O)O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 125000000962 organic group Chemical group 0.000 description 1
- IPCSVZSSVZVIGE-UHFFFAOYSA-N palmitic acid group Chemical group C(CCCCCCCCCCCCCCC)(=O)O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 1
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N pentanoic acid group Chemical class C(CCCC)(=O)O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- WLJVXDMOQOGPHL-UHFFFAOYSA-N phenylacetic acid Chemical compound OC(=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-UHFFFAOYSA-N 0.000 description 1
- 239000004014 plasticizer Substances 0.000 description 1
- 229940100467 polyvinyl acetate phthalate Drugs 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- UQDJGEHQDNVPGU-UHFFFAOYSA-N serine phosphoethanolamine Chemical compound [NH3+]CCOP([O-])(=O)OCC([NH3+])C([O-])=O UQDJGEHQDNVPGU-UHFFFAOYSA-N 0.000 description 1
- ZLGIYFNHBLSMPS-ATJNOEHPSA-N shellac Chemical compound OCCCCCC(O)C(O)CCCCCCCC(O)=O.C1C23[C@H](C(O)=O)CCC2[C@](C)(CO)[C@@H]1C(C(O)=O)=C[C@@H]3O ZLGIYFNHBLSMPS-ATJNOEHPSA-N 0.000 description 1
- 239000004208 shellac Substances 0.000 description 1
- 229940113147 shellac Drugs 0.000 description 1
- 235000013874 shellac Nutrition 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 235000015424 sodium Nutrition 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 235000010413 sodium alginate Nutrition 0.000 description 1
- 239000000661 sodium alginate Substances 0.000 description 1
- 229940005550 sodium alginate Drugs 0.000 description 1
- 229910001467 sodium calcium phosphate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 108010048412 somatotropin (177-191) Proteins 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 230000003019 stabilising effect Effects 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 229940032147 starch Drugs 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 238000009495 sugar coating Methods 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- HNKJADCVZUBCPG-UHFFFAOYSA-N thioanisole Chemical compound CSC1=CC=CC=C1 HNKJADCVZUBCPG-UHFFFAOYSA-N 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- XOOUIPVCVHRTMJ-UHFFFAOYSA-L zinc stearate Chemical compound [Zn+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O XOOUIPVCVHRTMJ-UHFFFAOYSA-L 0.000 description 1
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
- A61K38/22—Hormones
- A61K38/27—Growth hormone [GH], i.e. somatotropin
-
- 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
-
- 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
- A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
- A61K9/145—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic compounds
-
- 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
- A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
- A61K9/146—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic macromolecular compounds
-
- 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
- A61K9/20—Pills, tablets, discs, rods
- A61K9/2004—Excipients; Inactive ingredients
- A61K9/2013—Organic compounds, e.g. phospholipids, fats
Definitions
- This invention relates to pharmaceutical or veterinary formulations comprising a peptide as an active ingredient and methods for improving the stability of peptides in formulations, particularly where said peptides are orally available .
- Peptides are susceptible to aggregation and/or chemical degradation when stored in an aqueous solution for extended periods of time. This tendency of peptides to aggregate or degrade is generally characterized as "instability" . To extend the long term storage of peptides are often dried (for example by spray-drying or freeze-drying) for long-term storage.
- the present invention in a first aspect provides a pharmaceutical or veterinary formulation comprising as an active ingredient a peptide susceptible to degradation by acid hydrolysis and amount of free base basic amino acid sufficient to reduce the acid hydrolysis of the peptide.
- the invention provides a method of increasing the shelf-life of a pharmaceutical or veterinary formulation comprising as an active ingredient a peptide susceptible to degradation by acid hydrolysis comprising the step of adding a free base basic amino acid to the formulation.
- the peptide and the amino acid are present in a dry form and particularly as a dry powder.
- AOD9604 The applicant has studied the storage stability of their orally available peptide, AOD9604, described in International Patent application PCT/AU98/00724 and published as WO99/12969 (herein incorporated by reference) .
- Lyophilised pharmaceutical formulations of AOD9604, Tyr-hGH 177-191, amino acid sequence NH 2 -Tyr-Leu- Arg-Ile-Val-Gln-Cys-Arg-Ser-Val-Gln-Gly-Ser-Cys-Gly-Phe- COOH (SEQ ID NO: 1) are unstable at room temperature in dry form after three months .
- AOD9604 is manufactured using conventional solid phase chemical synthesis and its final form is as a lyophilized hydrochloride salt.
- HCl is present in the peptide formulation and that this HCl causes the AOD9604 to degrade, via acid hydrolysis.
- Such peptides include the applicant's orally available conotoxin peptide ACV-3 as described in their PCT application PCT/AU2006/001098 and having the amino acid sequence provided as SEQ ID NO: 2.
- ACV-3 Ac-Tyr-Leu-Arg-Ile-Val-Gly-Cys-Cys-Ser-Asp-Pro-Arg- Cys-Asn-Tyr-Asp-His-Pro-Glu-Ile-Cys-NH 2 (SEQ ID NO: 2)
- the formulation is a solid dosage form, in particular for oral use, such as a tablet or a capsule and the peptide is AOD9604 or ACV-3.
- the solid dosage form may also be for administration to the respiratory tract .
- the present invention provides a tablet comprising AOD9604 or ACV-3 as active ingredient and a free base basic amino acid.
- the present invention provides a capsule comprising AOD9604 or ACV-3 as active ingredient and a free base basic amino acid.
- the present invention provides a method of making a pharmaceutical or veterinary formulation comprising the step of adding to a peptide susceptible to degradation by acid hydrolysis a suitable amount of a free base basic amino acid to reduce the acid hydrolysis of the peptide .
- the peptide and amino acid are provided in dry form.
- the peptide and amino acid are dry blended to homogeneity to form the formulation.
- Figure 1 shows the four formulations used to assess the effect of arginine on stability in capsule formulations.
- Figure 2 shows a graph plotting the 6 month stability of various formulations containing Arginine, stored at 25 degrees Celsius and 60% relative humidity.
- Figure 3 shows a graph plotting the 6 month stability of various formulations containing Arginine, stored at 40 degrees Celsius and 75% relative humidity.
- Figure 4 shows a graph plotting the 6 month stability of various formulations containing Arginine, stored at 60 degrees Celsius.
- Figure 5 shows the percentage of total related substances present in each formulation initially, and at 1, 3, and 6 month time points.
- Figure 6 shows a graph plotting the 12 month stability of various capsule formulations containing Arginine, stored at 25 degrees Celsius and 60% relative humidity.
- Figure 7 shows a graph plotting the 12 month stability of various capsule formulations containing Arginine, stored at 40 degrees Celsius and 75% relative humidity.
- Figure 8 shows a graph plotting the 12 month stability of various capsule formulations containing Arginine, stored at 60 degrees Celsius.
- Figure 9 shows the formulations used to assess the effect of Arginine on stability in tablet formulations.
- Figure 10 shows a graph plotting the 9 month stability of various tablet formulations containing Arginine, stored at 4, 25 and 40 degrees Celsius (TEAP method) .
- Figure 11 shows a graph plotting the 9 month stability of various tablet formulations containing Arginine, stored at 4, 25 and 40 degrees Celsius (TFA method) .
- the free base basic amino acid may be Arginine, Lysine or Histidine or corresponding non-conventional amino acids, for example 5-Hydroxylysine, Homoarginine, Ornithine or Citrulline.
- a D-amino acid may be used instead of the corresponding L-amino acid, any amino acid may be N- methylated, or the N-terminus may be acetylated.
- Free-base Arginine is the preferred basic amino acid as it has the highest pKa and accordingly is proposed to be the most effective at reducing the acid hydrolysis of the peptide. Additionally L-Arginine is readily available, has no adverse biological activity and is non-toxic.
- the basic amino acid is in the L-form.
- the basic amino acid is L-Arginine in free base form.
- the formulation is formulated for oral administration.
- Administration to the respiratory tract is also contemplated.
- the formulation is in a capsule or formed into a tablet or is provided as a dry powder for use in a dry powder inhaler.
- the peptide and the amino acid are preferably in dry form.
- the formulation may also further comprise one or more dry excipients .
- Dry form as used herein means ingredients that are in solid form, including flowable solids which have some liquid-like properties. These generally contain less than 6% moisture. Dry form peptide formulations include freeze dried, lyophilised or desiccated peptide formulations.
- the active peptide may be any peptide which is susceptible to acid hydrolysis.
- the present invention is particularly suited to peptides provided in acid salt form. It is more particularly suited to peptides that are for oral administration, particularly those peptides that are orally bioavailable, such as AOD9604, ACV-3 and certain cyclosporins .
- the invention may be particularly applicable to peptides that are less than 50 amino acids long, and more particularly applicable to peptides that are less than 25 amino acids long.
- the peptide is an analogue of the C terminus of human growth hormone of between 8 and 30 amino acids in length.
- the peptide is active to increase lipolysis and/or decrease lipogenesis in adipocytes.
- the active peptide is AOD9604 or a lipid metabolising subfragment or analogue thereof.
- the active peptide is ACV-3 or an active subfragment or analogue thereof.
- the active peptide may be in the form of a pharmaceutically acceptable derivative, preferably a pharmaceutically acceptable salt.
- pharmaceutically acceptable salts include salts of pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, ammonium and alkylammonium; acid addition salts of pharmaceutically acceptable inorganic acids such as hydrochloric, orthophosphoric, sulphuric, phosphoric, nitric, carbonic, boric, sulfamic and hydrobromic acids; or salts of pharmaceutically acceptable organic acids such as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulphonic, trihalomethanesulphonic , toluenesulphonic , benzenesulphonic, salicylic, sulphanilic, aspartic, glutamic,
- the ratio of basic amino acid to peptide in the formulation is more than 2:1 by weight.
- the ratio of basic amino acid to peptide in the formulation is more than 4:1 by weight.
- the ratio of basic amino acid to peptide in the formulation is more than 8:1 by weight.
- Excess basic amino acid may be used in the formulation (as high as a ratio of 20:1 or more by weight) provided the basic amino acid is well tolerated.
- the pharmaceutical composition described in this invention could be used to treat any mammal .
- the animal may be a human, or may be a domestic or companion animal. While it is particularly contemplated that the present invention is used in medical treatment of humans, it is also applicable to veterinary treatment, including treatment of companion animals such as dogs and cats, and domestic animals such as horses, cattle and sheep, or zoo animals such as non- human primates, felids, canids, bovids, and ungulates.
- the mammal is a human.
- the human may be a child or an adult.
- peptide in this specification includes polypeptides of any amino acid length, including proteins, unless specifically restricted.
- oral delivery or “oral administration” are intended to encompass any administration or delivery to the GI tract and includes administration directly to the oropharyngeal cavity, and administration via the mouth in which the actual absorption of the peptide or polypeptide takes place in the gastrointestinal tract, including the stomach, small intestine, or large intestine.
- Oral administration as used herein encompasses sublingual administration
- An orally available peptide as defined herein is a peptide that is suitable for oral administration.
- Oral delivery and oral administration may be used interchangeably herein.
- Formulations for oral use include tablets containing the active compound in a mixture with non-toxic pharmaceutically acceptable excipients.
- These- excipients may be, for example, inert diluents or fillers ⁇ e.g., sucrose, sorbitol, sugar, mannitol, mirocrystalline cellulose, starches including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate or sodium phosphate) ; granulating and disintegrating agents ⁇ e.g., cellulose derivatives including microcrystalline cellulose, starches including potato starch, croscarmellose sodium, alginates or alginic acid); binding agents [e.g.
- Other pharmaceutically acceptable excipients can be colourants, flavouring agents, plasticisers, humectants, buffering agents and the like.
- the tablets may be uncoated or they may be coated by known techniques, optionally to delay disintegration and absorption in the gastrointestinal tract and thereby providing a sustained action over a longer period.
- the coating may be adapted to release the active compound in a predetermined pattern (e.g., in order to achieve a controlled release formulation) or it may be adapted not to release the active compound until after passage of the stomach (enteric coating) .
- the coating may be a sugar coating, a film coating (e.g., based on hydroxypropyl methylcellulose, methylcellulose, methyl hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, acrylate copolymers, polyethylene glycols and/or polyvinylpyrrolidone) , or an enteric coating (e.g., based on methacrylic acid copolymer, cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, polyvinyl acetate phthalate, shellac and/or ethylcellulose) .
- a time delay material such as, glyceryl monostearate or glyceryl distearate may be employed.
- the solid tablet compositions may include a coating adapted to protect the composition from unwanted chemical changes, (e.g., chemical degradation prior to the release of the active compound) .
- Formulations for oral use may also be presented as chewing tablets or as hard gelatin capsules wherein the active compound is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin) .
- an inert solid diluent e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin
- Powders and granulates may be prepared using the ingredients mentioned above under tablets and capsules in a conventional manner using, e.g, a mixer, a fluid bed apparatus or a spray drying equipment .
- the peptide may also be administered to the respiratory tract, for example by intranasal administration, by methods and formulations employed in the art for administration to the respiratory tract.
- the peptide may be administered to the respiratory tract in the form of a dry powder .
- Administration to the respiratory tract may be achieved by means of a dry powder inhaler (DPI) .
- DPI dry powder inhaler
- the powder composition may be presented in unit dose form, for example in capsules or cartridges of eg. gelatin, or blister packs from which the powder may be administered by means of an inhaler.
- the active compound will generally have a small particle size, for example of the order of 5 microns or less .
- a particle size may be obtained by means known in the art, for example by micronisation.
- formulations adapted to give sustained release of the active compound may be employed.
- the active compound may be administered by oral inhalation as a free-flow powder via a "Diskhaler” (trade mark of Glaxo Wellcome pic or a meter dose aerosol inhaler.
- a "Diskhaler” trade mark of Glaxo Wellcome pic or a meter dose aerosol inhaler.
- free base is intended to mean the free amino acid, not in the form of a salt.
- the formulation may comprise pharmaceutically acceptable carrier and, or excipients.
- Methods and pharmaceutical carriers for preparation of pharmaceutical compositions are well known in the art, as set out in textbooks such as Remington's Pharmaceutical Sciences, 20th Edition, Williams and Williams, Pennsylvania, USA (2000) .
- the formulation may further contain or be used in conjunction with other pharmaceutical preparations.
- a "pharmaceutically acceptable carrier” is a pharmaceutically acceptable solvent, a diluent, a suspending agent, excipient or vehicle for delivering the peptide to the subject.
- the formulation preferably comprises a bulking agent (which acts as a diluent and disintegrant , for example ProSolv HD90TM) , if necessary a flow enhancer, such as fumed silica (for example Cab-O-silTM) , and if necessary a lubricant, for example magnesium sterate, in addition to the peptide and free base amino acid.
- a bulking agent which acts as a diluent and disintegrant , for example ProSolv HD90TM
- a flow enhancer such as fumed silica (for example Cab-O-silTM)
- a lubricant for example magnesium sterate
- Preferred AOD tablet formulations are provided in Figure 9.
- the lubricant need not be present.
- the components of the formulations will depend upon which peptide is included. However it is envisaged that the presence of free base amino acid in a ratio of 2:1 to 10:1 and possibly up to 20:1 or more will reduce acid hydrolysis of the peptide.
- the formulations are preferably prepared and administered in dosage units.
- Solid dosage units include tablets, capsules and suppositories.
- different daily doses can be used depending on activity of the compound, manner of administration, nature and severity of the disorder, age and body weight of the subject. Under certain circumstances, however, higher or lower daily doses may be appropriate.
- the administration of the daily dose can be carried out both by single administration in the form of an individual dose unit or else several smaller dose units and also by multiple administration of subdivided doses at specific intervals.
- the formulations according to the invention may be administered in a therapeutically effective dose. Amounts effective for this use will, of course, depend on the severity of the disease and the weight and general state of the subject. Typically, dosages used in vitro may provide useful guidance in the amounts useful for in situ administration of the pharmaceutical composition, and animal models may be used to determine effective dosages for treatment of the cytotoxic side effects. Various considerations are described, e.g. in Langer, Science, 249: 1527, (1990) .
- the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease undergoing therapy.
- a peptide includes a plurality of such peptides
- a reference to “an amino acid” is a reference to one or more amino acids.
- the present invention employs conventional chemistry and protein chemistry within the capacity of those skilled in the art. Such techniques are well known to the skilled worker, and are explained fully in the literature. See, for example, Coligan, Dunn, Ploegh, Speicher and Wingfield: "Current protocols in Protein Science” (1999) Volumes I and II (John Wiley & Sons Inc . ) .
- AOD9604 has the sequence provided as SEQ ID NO" 1.
- Wang resin (0.625 g, 0.5 mmol) was placed in a 10 ml reaction vessel .
- Fmoc-L-phenylalanine Fmoc-Phe, 0.388 g, 1.0 mmol
- NMP-DCM 1:5, v/v
- DIC 0.135 g, 1.0 mmol
- 1.0 ml NMP 4-dimethylaminopyridine (DMAP, 0.074g, 0.06 mmol) in 0.6 ml DMF was added.
- DMAP 4-dimethylaminopyridine
- the reaction in the solution was allowed to continue for 68 minutes at room temperature. The solution was then drained, the resin was washed thoroughly with NMP (4 ml x 3) and OCM (4 ml x 3) .
- the Fmoc-Phe-Wang Resin complex was dried in vacuo overnight to yield 0.781 g of material.
- the coupling level of amino acid to resin was determined to be 0.80 mmol/g resin by using spectrophotometry measurement of the Fmoc- piperidine adduct . Step 3.
- Fmoc-Phe-Wang Resin (0.263 g, 0.20 tnmol) was placed in the 10 ml reaction vessel. DMF (8 ml) was added to wash and swell the resin by stirring for 2 minutes. The solution was then drained from the reaction vessel.
- Fmoc-glycine (Fmoc-Gly, 0.238 g, 0.8 mmol) , HOBt (108 mg; 0.8 mmol) and DIC (128 ⁇ l ; 0.8 mmol) were added to a 10 ml test tube containing 2 ml DMF. The mixture was stirred for 10 minutes to start the activation of amino acid. The solution was then added to the resin which originally was placed in the reaction vessel . The resulting mixture was stirred for 1.5 hours or until a negative ninhydrin test was obtained. The solution was then drained from the reaction vessel . Step 7 .
- Fmoc-Cys Acm) Fmoc-Ser (t-Bu) Fmoc-Gly Fmoc-Glu (O-tBu) Fmoc-Val Fmoc-Ser (t-Bu) Fmoc-Arg (Pmc) Fmoc-Cys (Acm) Emoc-Gln Fmoc-Val Fmoc-lle Fmoc-Arg (Pmc) Fmoc-Leu Fmoc-Tyr (t-Bu)
- the reaction vessel containing the peptide-resin was then placed in a desiccator and dried overnight under vacuum.
- the yield of peptide-resin was 0.635 g.
- the dried peptide-resin was removed from the reaction vessel and placed in a 50 ml round-bottom flask containing a magnetic stirring bar.
- the cleavage of the peptide from the resin with TFA was carried out with the following procedure: A scavenger solution, containing 0.75 g phenol, 0.5 ml H20, 0.5 ml thioanisole, and 0.25 ml ethanedithio, was added to the round-bottom flask. The resulting mixture was stirred for 5 minutes. 10 ml TFA was added drop by drop into the flask while kept stirring vigorously. The resulting mixture was stirred for 2.5 hours at room temperature .
- the mixture was filtered through a medium-porosity filter, fritted glass funnel .
- the TFA-peptide solution was sucked into another 500 ml round-bottom flask containing 200 ml cold diethyl ether by applying vacuum. Peptide was allowed to be precipitated in the ether solution at 4OC overnight, then collected by filtering the mixture through a fine- porosity, fritted glass funnel.
- the peptide pellet on the filter was washed with cold ether (10 ml x 3) to remove the scavenger.
- the peptide pellet was then dissolved with 25% aqueous acetic acid and then lyophilized to yield the crude peptide (about 400 mg dry weight, purity -80%) .
- the crude peptide was purified by reversed-phase high performance liquid chromatography (RP-HPLC) . Purification was carried out on a preparative 21.2 x 250mm Supelcosil PLC-18 (octadecyl, C18) column (120 Angstrom pore size, 12 ⁇ m particle size, 190 m2/g surface area; Supelco, Beliefonte, PA, U.S.A.) at 5.0 ml/mm flow rate at room temperature. A linear gradient program was utilized , where solvent A was water with 0.1 % TEA, and solvent B was acetonitrile-water (50/50: v/v, containing 0.1% TEA). The gradient was developed from 20 to 100% over 80mm.
- RP-HPLC reversed-phase high performance liquid chromatography
- AOD9604 acetate salt was dissolved in 1OmM HCl and relyophilised to yield AOD9604 Hydrochloride salt .
- the final powder blend was encapsulated using the Dott Bonopace encapsulator fitted with size 0 change parts.
- Formulation 4 was hazy and did not fully dissolve in solution at initial testing point. Solution remained hazy despite additional sonication/shaking and remained cloudy for up to 5 days after preparation. This probably accounts for the lower levels of AOD9604 detected at the initial timepoint .
- Tablets comprising ltng, 0.5 mg, 0.25 mg and 0 mg of AOD9604 were formulated with varying amounts of Arginine were made. Stability analysis on the tablet formulations were performed to determine the optimum level of Arginine that improves the stability characteristics of the formulation.
- Direct Compression Tablets The ProSolv, L-Arginine, Cab- O-sil and AOD9604 were accurately weighed and placed in a V-blender and blended for between 5 and 20 minutes. The Mg Stearate was added to the shell and blended for a further 5 - 20 minutes. The total blend time was 25 minutes . The blend was placed in the tablet press hopper and adjusted for weight and thickness settings to yield 100 mg tablets with thickness of approximately 4.2mm.
- the graphs show the results of HPLC methods used to determine the purity of AOD . Two methods are required in case an impurity is eluting at the same time as the main peak, this would be less likely to occur under two differenent sets of conditions.
- TFA refers to Trifluoro acetic acid
- TEAP refers to Triethylammonium phosphate .
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Endocrinology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Medicinal Preparation (AREA)
Abstract
The present invention provides a pharmaceutical or veterinary formulation comprising as an active ingredient a peptide susceptible to degradation by acid hydrolysis and amount of free base basic amino acid sufficient to reduce the acid hydrolysis of the peptide. Also provided is a method of increasing the shelf life of peptide formulations by adding a free base basic amino acid.
Description
Stabilisation of peptides with basic amino acids
This invention relates to pharmaceutical or veterinary formulations comprising a peptide as an active ingredient and methods for improving the stability of peptides in formulations, particularly where said peptides are orally available .
BACKGROUND
Peptides are susceptible to aggregation and/or chemical degradation when stored in an aqueous solution for extended periods of time. This tendency of peptides to aggregate or degrade is generally characterized as "instability" . To extend the long term storage of peptides are often dried (for example by spray-drying or freeze-drying) for long-term storage.
Even dried peptide formulations tend to be unstable at room temperature. Degradation products formed from deamidation, oxidation and cleavage are often generated, limiting the room temperature shelf life. For this reason, a stable formulation is highly desirable for a peptide drug to enable distribution and storage without refrigeration, particularly where large market indications with oral formulations are concerned.
It is an aim of a preferred embodiment of the present invention to improve the storage stability of peptides for pharmaceutical or veterinary use and to provide peptide formulations with improved stability.
SUMMARY
The present invention in a first aspect provides a pharmaceutical or veterinary formulation comprising as an active ingredient a peptide susceptible to degradation by
acid hydrolysis and amount of free base basic amino acid sufficient to reduce the acid hydrolysis of the peptide.
In a second aspect the invention provides a method of increasing the shelf-life of a pharmaceutical or veterinary formulation comprising as an active ingredient a peptide susceptible to degradation by acid hydrolysis comprising the step of adding a free base basic amino acid to the formulation.
Preferably the peptide and the amino acid are present in a dry form and particularly as a dry powder.
The applicant has studied the storage stability of their orally available peptide, AOD9604, described in International Patent application PCT/AU98/00724 and published as WO99/12969 (herein incorporated by reference) . Lyophilised pharmaceutical formulations of AOD9604, Tyr-hGH 177-191, amino acid sequence NH2-Tyr-Leu- Arg-Ile-Val-Gln-Cys-Arg-Ser-Val-Gln-Gly-Ser-Cys-Gly-Phe- COOH (SEQ ID NO: 1) are unstable at room temperature in dry form after three months .
For clinical development AOD9604 is manufactured using conventional solid phase chemical synthesis and its final form is as a lyophilized hydrochloride salt. On further investigation the applicant determined, that when formulated in a PEG/Pearlitol capsule, HCl is present in the peptide formulation and that this HCl causes the AOD9604 to degrade, via acid hydrolysis. The applicant determined that the main degradation pathway is initiated by the deamidation of the glutamine residue of the peptide .
The applicant then determined that the addition of a free base basic amino acid to the formulation of AOD9604 increased the stability of the formulation against acid
hydrolysis, extending the room temperature stability from 3 months to more than 12 months.
The applicant envisages that the stabilising effect of the basic amino acid can be used to stabilise other peptide formulations which are susceptible to acid hydrolysis. Such peptides include the applicant's orally available conotoxin peptide ACV-3 as described in their PCT application PCT/AU2006/001098 and having the amino acid sequence provided as SEQ ID NO: 2.
ACV-3 : Ac-Tyr-Leu-Arg-Ile-Val-Gly-Cys-Cys-Ser-Asp-Pro-Arg- Cys-Asn-Tyr-Asp-His-Pro-Glu-Ile-Cys-NH2 (SEQ ID NO: 2)
In one embodiment, the formulation is a solid dosage form, in particular for oral use, such as a tablet or a capsule and the peptide is AOD9604 or ACV-3. The solid dosage form may also be for administration to the respiratory tract .
In a third aspect the present invention provides a tablet comprising AOD9604 or ACV-3 as active ingredient and a free base basic amino acid.
In the fourth aspect the present invention provides a capsule comprising AOD9604 or ACV-3 as active ingredient and a free base basic amino acid.
In a fifth aspect the present invention provides a method of making a pharmaceutical or veterinary formulation comprising the step of adding to a peptide susceptible to degradation by acid hydrolysis a suitable amount of a free base basic amino acid to reduce the acid hydrolysis of the peptide .
Preferably the peptide and amino acid are provided in dry form. Preferably the peptide and amino acid are dry
blended to homogeneity to form the formulation.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1 shows the four formulations used to assess the effect of arginine on stability in capsule formulations.
Figure 2 shows a graph plotting the 6 month stability of various formulations containing Arginine, stored at 25 degrees Celsius and 60% relative humidity.
Figure 3 shows a graph plotting the 6 month stability of various formulations containing Arginine, stored at 40 degrees Celsius and 75% relative humidity.
Figure 4 shows a graph plotting the 6 month stability of various formulations containing Arginine, stored at 60 degrees Celsius.
Figure 5 shows the percentage of total related substances present in each formulation initially, and at 1, 3, and 6 month time points.
Figure 6 shows a graph plotting the 12 month stability of various capsule formulations containing Arginine, stored at 25 degrees Celsius and 60% relative humidity.
Figure 7 shows a graph plotting the 12 month stability of various capsule formulations containing Arginine, stored at 40 degrees Celsius and 75% relative humidity.
Figure 8 shows a graph plotting the 12 month stability of various capsule formulations containing Arginine, stored at 60 degrees Celsius.
Figure 9 shows the formulations used to assess the effect of Arginine on stability in tablet formulations.
Figure 10 shows a graph plotting the 9 month stability of various tablet formulations containing Arginine, stored at 4, 25 and 40 degrees Celsius (TEAP method) .
Figure 11 shows a graph plotting the 9 month stability of various tablet formulations containing Arginine, stored at 4, 25 and 40 degrees Celsius (TFA method) .
DETAILED DESCRIPTION
In a study of the stability of the dried formulations of AOD9604, four different formulations of capsules containing AOD9604 were manufactured containing varying amounts of free base Arginine as basic amino acid. Stability analysis on the capsule formulations were performed to determine the optimum level of Arginine that improves the stability characteristics of the formulation.
Comparison of the stability data shows that the formulations containing the most Arginine were the most stable, achieving a loss in potency of only approximately 20% with respect to initial value even after storage at elevated temperatures over a 6 month period. Physical decomposition of the formulations stored at 60°C was also observed in formulations without Arginine, or with low levels of Arginine, with no physical decomposition detected in the formulation containing the highest level of Arginine .
Assay results indicate a degeneration of AOD9604 over time at elevated temperature and humidity conditions, with less degradation occurring in the formulations containing higher levels of Arginine . Similarly, the levels of related substances increased over time with decreasing Arginine levels, with the formulation containing no Arginine totally degrading within 3 months at 6O0C.
Therefore, it is evident that there is a direct correlation between levels of Arginine and stability of AOD9604 in capsule formulations of AOD9604, with the optimum level being 40 - 80 mg per capsule (11.5 - 23%w/w or a ratio of 8:1 Arginine to peptide (w/w) ) . Arginine appears to contribute both to overall stability and initial release of the active ingredient into solution with higher levels demonstrating rapid release of the drug.
Further experiments showed that there is a direct correlation between levels of Arginine and stability of AOD9604 in tablet formulations of AOD9604.
Whilst all the examples have been carried out using Arginine as free base basic amino acid it would be immediately apparent to those skilled in the art of pharmaceutical peptide formulation that other basic amino acids could be used.
The free base basic amino acid may be Arginine, Lysine or Histidine or corresponding non-conventional amino acids, for example 5-Hydroxylysine, Homoarginine, Ornithine or Citrulline. A D-amino acid may be used instead of the corresponding L-amino acid, any amino acid may be N- methylated, or the N-terminus may be acetylated.
Free-base Arginine is the preferred basic amino acid as it
has the highest pKa and accordingly is proposed to be the most effective at reducing the acid hydrolysis of the peptide. Additionally L-Arginine is readily available, has no adverse biological activity and is non-toxic.
Preferably the basic amino acid is in the L-form. Preferably the basic amino acid is L-Arginine in free base form.
Preferably the formulation is formulated for oral administration. Administration to the respiratory tract is also contemplated.
Preferably, the formulation is in a capsule or formed into a tablet or is provided as a dry powder for use in a dry powder inhaler.
The peptide and the amino acid are preferably in dry form. The formulation may also further comprise one or more dry excipients .
Dry form as used herein means ingredients that are in solid form, including flowable solids which have some liquid-like properties. These generally contain less than 6% moisture. Dry form peptide formulations include freeze dried, lyophilised or desiccated peptide formulations.
The active peptide may be any peptide which is susceptible to acid hydrolysis. The present invention is particularly suited to peptides provided in acid salt form. It is more particularly suited to peptides that are for oral administration, particularly those peptides that are orally bioavailable, such as AOD9604, ACV-3 and certain cyclosporins .
The invention may be particularly applicable to peptides that are less than 50 amino acids long, and more
particularly applicable to peptides that are less than 25 amino acids long.
In a preferred embodiment, the peptide is an analogue of the C terminus of human growth hormone of between 8 and 30 amino acids in length.
Preferably, the peptide is active to increase lipolysis and/or decrease lipogenesis in adipocytes.
Preferably, the active peptide is AOD9604 or a lipid metabolising subfragment or analogue thereof.
In another embodiment the active peptide is ACV-3 or an active subfragment or analogue thereof.
It will be appreciated that the active peptide may be in the form of a pharmaceutically acceptable derivative, preferably a pharmaceutically acceptable salt. Examples of pharmaceutically acceptable salts include salts of pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, ammonium and alkylammonium; acid addition salts of pharmaceutically acceptable inorganic acids such as hydrochloric, orthophosphoric, sulphuric, phosphoric, nitric, carbonic, boric, sulfamic and hydrobromic acids; or salts of pharmaceutically acceptable organic acids such as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulphonic, trihalomethanesulphonic , toluenesulphonic , benzenesulphonic, salicylic, sulphanilic, aspartic, glutamic, edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic, ascorbic and valeric acids. Salts of amine groups may also comprise quaternary ammonium salts in which the amino nitrogen atom carries a suitable organic group such as an alkyl, alkenyl, alkynyl or
aralkyl moiety.
Preferably, the ratio of basic amino acid to peptide in the formulation is more than 2:1 by weight.
More preferably, the ratio of basic amino acid to peptide in the formulation is more than 4:1 by weight.
Most preferably, the ratio of basic amino acid to peptide in the formulation is more than 8:1 by weight.
Excess basic amino acid may be used in the formulation (as high as a ratio of 20:1 or more by weight) provided the basic amino acid is well tolerated.
The pharmaceutical composition described in this invention could be used to treat any mammal . The animal may be a human, or may be a domestic or companion animal. While it is particularly contemplated that the present invention is used in medical treatment of humans, it is also applicable to veterinary treatment, including treatment of companion animals such as dogs and cats, and domestic animals such as horses, cattle and sheep, or zoo animals such as non- human primates, felids, canids, bovids, and ungulates.
Preferably the mammal is a human. The human may be a child or an adult.
Use of the term "peptide" in this specification includes polypeptides of any amino acid length, including proteins, unless specifically restricted.
As referred to herein "oral delivery" or "oral administration" are intended to encompass any administration or delivery to the GI tract and includes administration directly to the oropharyngeal cavity, and administration via the mouth in which the actual
absorption of the peptide or polypeptide takes place in the gastrointestinal tract, including the stomach, small intestine, or large intestine. Oral administration as used herein encompasses sublingual administration
(administration by application under the tongue of the recipient, representing one form of administration via the oropharyngeal cavity) and buccal administration
(administration of a dosage form between the teeth and the cheek of the recipient) .
An orally available peptide as defined herein is a peptide that is suitable for oral administration.
Oral delivery and oral administration may be used interchangeably herein.
Formulations for oral use include tablets containing the active compound in a mixture with non-toxic pharmaceutically acceptable excipients. These- excipients may be, for example, inert diluents or fillers {e.g., sucrose, sorbitol, sugar, mannitol, mirocrystalline cellulose, starches including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate or sodium phosphate) ; granulating and disintegrating agents {e.g., cellulose derivatives including microcrystalline cellulose, starches including potato starch, croscarmellose sodium, alginates or alginic acid); binding agents [e.g. , sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminium silicate, carboxymethylcellulose sodium, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinylpyrrolidone or polyethylene glycol) ; and lubricating agents, glidants, and antiadhesives {e.g., magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenated vegetable oils or talc) . Other pharmaceutically acceptable excipients can
be colourants, flavouring agents, plasticisers, humectants, buffering agents and the like.
The tablets may be uncoated or they may be coated by known techniques, optionally to delay disintegration and absorption in the gastrointestinal tract and thereby providing a sustained action over a longer period. The coating may be adapted to release the active compound in a predetermined pattern (e.g., in order to achieve a controlled release formulation) or it may be adapted not to release the active compound until after passage of the stomach (enteric coating) . The coating may be a sugar coating, a film coating (e.g., based on hydroxypropyl methylcellulose, methylcellulose, methyl hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, acrylate copolymers, polyethylene glycols and/or polyvinylpyrrolidone) , or an enteric coating (e.g., based on methacrylic acid copolymer, cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, polyvinyl acetate phthalate, shellac and/or ethylcellulose) . Furthermore, a time delay material such as, glyceryl monostearate or glyceryl distearate may be employed.
The solid tablet compositions may include a coating adapted to protect the composition from unwanted chemical changes, (e.g., chemical degradation prior to the release of the active compound) .
Formulations for oral use may also be presented as chewing tablets or as hard gelatin capsules wherein the active compound is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin) . Powders and granulates may be prepared using the ingredients mentioned above under tablets and capsules in a
conventional manner using, e.g, a mixer, a fluid bed apparatus or a spray drying equipment .
The peptide may also be administered to the respiratory tract, for example by intranasal administration, by methods and formulations employed in the art for administration to the respiratory tract.
Thus in general the peptide may be administered to the respiratory tract in the form of a dry powder . Administration to the respiratory tract may be achieved by means of a dry powder inhaler (DPI) . The powder composition may be presented in unit dose form, for example in capsules or cartridges of eg. gelatin, or blister packs from which the powder may be administered by means of an inhaler.
In formulations intended for administration to the respiratory tract, including intranasal formulations, the active compound will generally have a small particle size, for example of the order of 5 microns or less . Such a particle size may be obtained by means known in the art, for example by micronisation.
When desired, formulations adapted to give sustained release of the active compound may be employed.
The active compound may be administered by oral inhalation as a free-flow powder via a "Diskhaler" (trade mark of Glaxo Wellcome pic or a meter dose aerosol inhaler.
As used herein "free base" is intended to mean the free amino acid, not in the form of a salt.
The formulation may comprise pharmaceutically acceptable carrier and, or excipients. Methods and pharmaceutical carriers for preparation of pharmaceutical compositions are well known in the art, as set out in textbooks such as
Remington's Pharmaceutical Sciences, 20th Edition, Williams and Williams, Pennsylvania, USA (2000) .
The formulation may further contain or be used in conjunction with other pharmaceutical preparations.
As used herein, a "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, a diluent, a suspending agent, excipient or vehicle for delivering the peptide to the subject.
The formulation preferably comprises a bulking agent (which acts as a diluent and disintegrant , for example ProSolv HD90™) , if necessary a flow enhancer, such as fumed silica (for example Cab-O-sil™) , and if necessary a lubricant, for example magnesium sterate, in addition to the peptide and free base amino acid.
Preferred AOD tablet formulations are provided in Figure 9. In a capsule formulation the lubricant need not be present. The components of the formulations will depend upon which peptide is included. However it is envisaged that the presence of free base amino acid in a ratio of 2:1 to 10:1 and possibly up to 20:1 or more will reduce acid hydrolysis of the peptide.
The formulations are preferably prepared and administered in dosage units. Solid dosage units include tablets, capsules and suppositories. For treatment of a subject, depending on activity of the compound, manner of administration, nature and severity of the disorder, age and body weight of the subject, different daily doses can be used. Under certain circumstances, however, higher or lower daily doses may be appropriate. The administration of the daily dose can be carried out both by single administration in the form of an individual dose unit or
else several smaller dose units and also by multiple administration of subdivided doses at specific intervals.
The formulations according to the invention may be administered in a therapeutically effective dose. Amounts effective for this use will, of course, depend on the severity of the disease and the weight and general state of the subject. Typically, dosages used in vitro may provide useful guidance in the amounts useful for in situ administration of the pharmaceutical composition, and animal models may be used to determine effective dosages for treatment of the cytotoxic side effects. Various considerations are described, e.g. in Langer, Science, 249: 1527, (1990) .
It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease undergoing therapy.
In the description of the invention and in the claims which follow, except where the context requires otherwise due to express language or necessary implication, the word "comprise" or variations such as "comprises" or "comprising" is used in an inclusive sense, i.e. to specify the presence of the stated features but not to preclude the presence or addition of further features in various embodiments of the invention.
As used herein, the singular forms "a" , "an" , and "the" include the corresponding plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to "a peptide" includes a plurality of such peptides, and a reference to "an amino acid" is a
reference to one or more amino acids.
Where a range of values is expressed, it will be clearly understood that this range encompasses the upper and lower limits of the range, and all values in between these limits .
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any materials and methods similar or equivalent to those described herein can be used to practice or test the present invention, the preferred materials and methods are described.
It is to be clearly understood that this invention is not limited to the particular materials and methods described herein, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and it is not intended to limit the scope of the present invention, which will be limited only by the appended claims.
Unless otherwise indicated, the present invention employs conventional chemistry and protein chemistry within the capacity of those skilled in the art. Such techniques are well known to the skilled worker, and are explained fully in the literature. See, for example, Coligan, Dunn, Ploegh, Speicher and Wingfield: "Current protocols in Protein Science" (1999) Volumes I and II (John Wiley & Sons Inc . ) .
The invention will now be described in detail by way of reference only to the following non-limiting examples.
EXAMPLES
SYNTHESIS OF HEXADECAPEPTIDE OF AMINO ACID RESIDUES 177-191 OF NATIVE HUMAN GROWTH HORONE, DESIGNATED HGH (177-191) WITH AN N-TERMINAL TYROSINE ELONGATION (REF NO: AOD9604)
AOD9604 has the sequence provided as SEQ ID NO" 1.
NHa-Tyr-Leu-Arg-Ile-Val-Gln-Cys-Arg-Ser-Val-Gln-Gly-Ser- Cys-GIy-Phe-COOH (SEQ ID NO: 1)
The following procedure was employed in preparing the hexadecapeptide, as previously described in WO99/12969.
Step 1.
Wang resin (0.625 g, 0.5 mmol) was placed in a 10 ml reaction vessel .
DCM (4 ml) was added to the reaction vessel. The Wang resin was washed with vigorous stirring for 2 minutes. The
DCM solution was then drained from the reaction vessel .
This washing was repeated twice.
Step 2.
Fmoc-L-phenylalanine (Fmoc-Phe, 0.388 g, 1.0 mmol) in 2.4 ml NMP-DCM (1:5, v/v) and DIC (0.135 g, 1.0 mmol) in 1.0 ml NMP were mixed in a reaction vessel for 10 minutes. To the mixture, 4-dimethylaminopyridine (DMAP, 0.074g, 0.06 mmol) in 0.6 ml DMF was added. The reaction in the solution was allowed to continue for 68 minutes at room temperature. The solution was then drained, the resin was washed thoroughly with NMP (4 ml x 3) and OCM (4 ml x 3) . The Fmoc-Phe-Wang Resin complex was dried in vacuo overnight to yield 0.781 g of material. The coupling level of amino acid to resin was determined to be 0.80 mmol/g resin by using spectrophotometry measurement of the Fmoc- piperidine adduct .
Step 3.
Fmoc-Phe-Wang Resin (0.263 g, 0.20 tnmol) was placed in the 10 ml reaction vessel. DMF (8 ml) was added to wash and swell the resin by stirring for 2 minutes. The solution was then drained from the reaction vessel.
Step 4.
A 25% piperidine/DMF solution (4 ml) was added to the reaction vessel . The resulting mixture was stirred for 2 minutes. The solution was drained from the reaction vessel. This deprotection procedure was repeated once but with prolonged stirring time (18 mm) . The solution was drained from the reaction vessel .
Step 5.
DMF (8 ml) was added to the reaction vessel. The resulting solution was stirred for 2 minutes. The solution was drained from the resin in the reaction vessel . This washing procedure was repeated twice. DMF (2 ml) was added to the reaction vessel to keep the resin swollen.
Step 6.
Fmoc-glycine (Fmoc-Gly, 0.238 g, 0.8 mmol) , HOBt (108 mg; 0.8 mmol) and DIC (128 μl ; 0.8 mmol) were added to a 10 ml test tube containing 2 ml DMF. The mixture was stirred for 10 minutes to start the activation of amino acid. The solution was then added to the resin which originally was placed in the reaction vessel . The resulting mixture was stirred for 1.5 hours or until a negative ninhydrin test was obtained. The solution was then drained from the reaction vessel .
Step 7 .
DMF (8 ml) was added to the reaction vessel. The resulting solution was vigorously stirred for 2 minutes. The solution was then drained from the reaction vessel . The washing procedure was repeated twice.
Steps 4 through 7 were then repeated employing the following order of amino acid:
Fmoc-Cys (Acm) Fmoc-Ser (t-Bu) Fmoc-Gly Fmoc-Glu (O-tBu) Fmoc-Val Fmoc-Ser (t-Bu) Fmoc-Arg (Pmc) Fmoc-Cys (Acm) Emoc-Gln Fmoc-Val Fmoc-lle Fmoc-Arg (Pmc) Fmoc-Leu Fmoc-Tyr (t-Bu)
After completion of the synthesis of the desired peptide- resin, the reaction vessel containing the peptide-resin was then placed in a desiccator and dried overnight under vacuum. The yield of peptide-resin was 0.635 g. The dried peptide-resin was removed from the reaction vessel and placed in a 50 ml round-bottom flask containing a magnetic stirring bar. The cleavage of the peptide from the resin with TFA was carried out with the following procedure: A scavenger solution, containing 0.75 g phenol, 0.5 ml H20,
0.5 ml thioanisole, and 0.25 ml ethanedithio, was added to the round-bottom flask. The resulting mixture was stirred for 5 minutes. 10 ml TFA was added drop by drop into the flask while kept stirring vigorously. The resulting mixture was stirred for 2.5 hours at room temperature .
The mixture was filtered through a medium-porosity filter, fritted glass funnel . The TFA-peptide solution was sucked into another 500 ml round-bottom flask containing 200 ml cold diethyl ether by applying vacuum. Peptide was allowed to be precipitated in the ether solution at 4OC overnight, then collected by filtering the mixture through a fine- porosity, fritted glass funnel. The peptide pellet on the filter was washed with cold ether (10 ml x 3) to remove the scavenger. The peptide pellet was then dissolved with 25% aqueous acetic acid and then lyophilized to yield the crude peptide (about 400 mg dry weight, purity -80%) .
The crude peptide was purified by reversed-phase high performance liquid chromatography (RP-HPLC) . Purification was carried out on a preparative 21.2 x 250mm Supelcosil PLC-18 (octadecyl, C18) column (120 Angstrom pore size, 12 μm particle size, 190 m2/g surface area; Supelco, Beliefonte, PA, U.S.A.) at 5.0 ml/mm flow rate at room temperature. A linear gradient program was utilized , where solvent A was water with 0.1 % TEA, and solvent B was acetonitrile-water (50/50: v/v, containing 0.1% TEA). The gradient was developed from 20 to 100% over 80mm. Separation profiles were recorded and analysed using a Perkin-Elmer LO-IQO integrator. The desired peptide component was eluted and collected with the Pharmacia Model PRAC-IQO automatic fraction collector (Uppsala, Sweden) . The fractions of identical component were
combined and lyophilized. The purified peptide (275 mg dry weight, purity >98%) , Cys(Acm)7-14 hexadecapeptide, was kept frozen at -20 OC.
For cyclisation of the disulfide bridge of the peptides, iodine oxidation in 80% aqueous acetic acid was used to remove the cysteine-protecting groups, Acm, and furnished the intramolecular disulfide bridge simultaneously. Cys (Acm) 7-14 pentadecapeptide (275 mg, 0.155 mmole) was dissolved in 50 ml 80% aqueous acetic acid. This solution was slowly added to a 250 ml round-bottom flask containing iodine (378 mg, 1.4 mmole) in 100 ml 80% aqueous acetic acid by stirring vigorously. Reaction was allowed to continue for 2 hours at room temperature and terminated by adding the ascorbic acid (Vitamin C) to the resulting solution. Liquid volume was then reduced by rotary evaporation and peptide recovered by lyophilization. The cyclised peptide was then purified by RP-HPLC as described in the purification of linear peptide. After lyophilization, 165 mg cyclic pentadecapeptide with 96% purity was yielded. The total yield of synthesis of AOD9604 was about 46%.
For formulation the AOD9604 acetate salt was dissolved in 1OmM HCl and relyophilised to yield AOD9604 Hydrochloride salt .
Capsule Formulations
Four different formulations of AOD9604 capsules containing varying amounts of Arginine were made. Stability analysis on the capsule formulations were performed to determine
the optimum level of Arginine that improves the stability characteristics of the formulation.
Four formulations were manufactured, each containing different amounts of free base Arginine as indicated in Figure 1 below.
All four formulations were prepared as follows:
1. All excipients were screened through a 40 mesh sieve
2. AOD9604 , PEG3350 and an equivalent amount of Pearlitol were added to the cube blender and mixed for 10 minutes
3. Resultant powder blend was screened through a 40 mesh screen and returned to the cube mixer
4. Arginine was added to the blend and mixed for a further 10 minutes
5. Resultant powder blend was again screened though a 40 mesh sieve and returned to the cube mixer
6. Remaining quantity of Pearlitol was added to the blend and mixed for 10 minutes
7.5 x 1 gram samples were taken to be tested for blend homogeneity 8. Balance of blend collected and stored at -200C awaiting homogeneity results.
Following receipt of satisfactory homogeneity results, the final powder blend was encapsulated using the Dott Bonopace encapsulator fitted with size 0 change parts.
Following homogeneity results, the four formulations were placed on a 6 month stability study to ascertain the correlation between Arginine levels and the stability of AOD9604 with respect to dissolution and drug release profiles. Capsules were stored at 25°C/60% relative humidity (RH) , 40°C/75%RH and 60°C with sampling points of 1 month, 3 months and 6 months.
The results obtained for the assay of each formulation at each stability time point and storage condition are presented in Figures 2 to 4 below.
Note: Formulation 4 was hazy and did not fully dissolve in solution at initial testing point. Solution remained hazy despite additional sonication/shaking and remained cloudy for up to 5 days after preparation. This probably accounts for the lower levels of AOD9604 detected at the initial timepoint .
Comparison of the stability suggests a correlation between Arginine levels and product stability, with Formulation 3 and Formulation 4 achieving a loss in potency of only approximately 20% with respect to initial value even after storage at elevated temperatures over a 6 month period.
Physical decomposition of the formulations stored at 60°G was also observed in Formulations 1 and 2 within 3 months of storage and to a lesser extent in Formulation 3, with 20% of capsule contents examined showing discolouration. No discolouration was observed in the contents of capsules containing Formulation 4.
Assay results indicate a degeneration of AOD9604 over time at elevated temperature and humidity conditions, with less degradation occurring in the formulations containing higher levels of Arginine. Similarly, related substances levels increased over time with decreasing Arginine levels, with Formulation 1 totally degrading within 3 months at 60°C. Formulation 4 appeared to be the most stable with respect to assay degradation and formation of related substances, see Figure 5.
From this experiment it is evident that there is a direct correlation between levels of Arginine and stability of
AOD9604 in capsule formulations, with the optimum level being 40 - 80 mg per capsule (4:1 - 8:1 Arginine to AOD9604 w/w) .
The stability study was continued for 12 months and the further results shown in Figures 6-8. The trend of reduction in stability of the AOD formulation is continued. After 12 months under 75% relative humidity and 60 degrees centigrade, the formulation comprising AOD9604 and 12% or 24% Arginine is shown to be stable.
Tablet Formulations
Tablets comprising ltng, 0.5 mg, 0.25 mg and 0 mg of AOD9604 were formulated with varying amounts of Arginine were made. Stability analysis on the tablet formulations were performed to determine the optimum level of Arginine that improves the stability characteristics of the formulation.
Four formulations were manufactured, each containing different amounts of free base Arginine as indicated in Figure 9 below.
Direct Compression Tablets: The ProSolv, L-Arginine, Cab- O-sil and AOD9604 were accurately weighed and placed in a V-blender and blended for between 5 and 20 minutes. The Mg Stearate was added to the shell and blended for a further 5 - 20 minutes. The total blend time was 25 minutes . The blend was placed in the tablet press hopper and adjusted for weight and thickness settings to yield 100 mg tablets with thickness of approximately 4.2mm.
The formulations were placed on a 24 month stability study. Tablets were stored at 4, 25 and 4O0C. So far sampling points of 1 month, 3 months, 6 months and 9
months have been taken.
The graphs show the results of HPLC methods used to determine the purity of AOD . Two methods are required in case an impurity is eluting at the same time as the main peak, this would be less likely to occur under two differenent sets of conditions. TFA refers to Trifluoro acetic acid, and TEAP refers to Triethylammonium phosphate .
The results obtained for the assay of each formulation at each stability time point and storage condition are presented in Figures 10 and 11 below.
From this experiment it is evident that there is a direct correlation between levels of Arginine and stability of AOD9604 in tablet formulations. It is anticipated that this correlation would also be seen with other basic amino acids .
Additionally it is also envisaged that the stabilizing effect of the basic amino acids would extend to other dried peptide formulations susceptible to acid hydrolysis.
Claims
1. A pharmaceutical or veterinary formulation comprising as an active ingredient a peptide susceptible to degradation by acid hydrolysis and amount of free base basic amino acid sufficient to reduce the acid hydrolysis of the peptide.
2. A formulation according to claim 1 in dry form.
3. A formulation according to claim 1 as a solid dosage unit .
4. A formulation according to claim 1 in which the basic amino acid is Arginine, Lysine, Histidine, Ornithine, or Citrulline .
5. A formulation according to claim 1 in which the basic amino acid is Arginine .
6. A formulation according to claim 1 in which the basic amino acid is an L amino acid.
7. A formulation according to any preceding claim formulated for oral administration.
8. A formulation according to any preceding claim in tablet form.
9. A formulation according to any one of claims 1 to 7 in capsule form.
10. A formulation according to any preceding claim in which the peptide is a peptide that is susceptible to acid hydrolysis .
11. A formulation according to claim 10 in which the peptide is in acid salt form.
12. A formulation according to claim 10 in which the peptide is less than 50 amino acids in length.
13. A formulation according to claim 10 in which the peptide is an analogue of the C terminus of human growth hormone of between 8 and 30 amino acids in length.
14. A formulation according to claim 13 in which the peptide is active to increase lipolysis and/or decrease lipogenesis in adipocytes.
15. A formulation according to claim 13 in which the peptide is AOD9604 or a lipid metabolising subfragment or analogue thereof .
16. A formulation according to claim 10 in which the peptide is ACV-3 or an active subfragment or analogue thereof .
17. A formulation according to claim 1 in which the ratio of free base basic amino acid to peptide is more than 2:1 by weight .
18. A formulation according to claim 1 in which the ratio of free base basic amino acid to peptide is more than 4:1 by weight .
19. A formulation according to claim 1 in which the ratio of free base basic amino acid to peptide is more than 8:1 by weight .
20. A method of increasing the shelf-life of a pharmaceutical or veterinary formulation comprising as an active ingredient a peptide susceptible to degradation by acid hydrolysis comprising the step of adding a free base basic amino acid to the formulation.
21. A method according to claim 20 in which the basic amino acid is Arginine, Lysine, Histidine, Ornthine, or Citrulline .
22. A method according to claim 20 in which the peptide is AOD9604 or a lipid metabolising subfragment or analogue thereof .
23. A formulation according to claim 20 in which the peptide is ACV-3 or an active subfragment or analogue thereof .
24. A method according to claim 20 in which the formulation is for oral administration.
25. A tablet comprising AOD9604 or a lipid metabolising subfragment or analogue thereof as active ingredient and a free base basic amino acid.
26. A capsule comprising AOD9604 or a lipid metabolising subfragment or analogue thereof as active ingredient and a free base basic amino acid.
27. A tablet comprising ACV-3 or an active subfragment or analogue thereof as active ingredient and a free base basic amino acid.
28. A capsule comprising ACV-3 or an active subfragment or analogue thereof as active ingredient and a free base basic amino acid.
29. A method of making a pharmaceutical or veterinary formulation comprising the step of adding to a peptide susceptible to acid hydrolysis a suitable amount of a free base basic amino acid to reduce the acid hydrolysis of the peptide .
30. A method according to claim 25 in which the peptide and amino acid are dry mixed.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU2005905247A AU2005905247A0 (en) | 2005-09-23 | Peptide fomulation | |
AU2005905247 | 2005-09-23 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2007033427A1 true WO2007033427A1 (en) | 2007-03-29 |
Family
ID=37888462
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/AU2006/001391 WO2007033427A1 (en) | 2005-09-23 | 2006-09-22 | Stabilisation of peptides with basic amino acids |
Country Status (1)
Country | Link |
---|---|
WO (1) | WO2007033427A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020012243A2 (en) | 2018-07-09 | 2020-01-16 | Abbott Laboratories Gmbh | Pharmaceutical composition containing a peptide |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4816568A (en) * | 1986-05-16 | 1989-03-28 | International Minerals & Chemical Corp. | Stabilization of growth hormones |
WO1993012812A1 (en) * | 1991-12-20 | 1993-07-08 | Novo Nordisk A/S | A stabilized pharmaceutical formulation comprising growth hormone and histidine |
WO1997039768A1 (en) * | 1996-04-24 | 1997-10-30 | Novo Nordisk A/S | A pharmaceutical formulation containing growth hormone, an amino acid and a non-ionic detergent |
US6013773A (en) * | 1996-02-02 | 2000-01-11 | Mitsui Chemicals, Inc. | Pharmaceutical preparation containing human growth hormone |
US6204036B1 (en) * | 1995-03-09 | 2001-03-20 | Aventis Behring Gmbh | Stable transglutaminase preparations and processes for their production |
WO2001049314A2 (en) * | 1999-12-30 | 2001-07-12 | Nps Allelix Corp. | Glp-2 formulations |
-
2006
- 2006-09-22 WO PCT/AU2006/001391 patent/WO2007033427A1/en active Application Filing
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4816568A (en) * | 1986-05-16 | 1989-03-28 | International Minerals & Chemical Corp. | Stabilization of growth hormones |
WO1993012812A1 (en) * | 1991-12-20 | 1993-07-08 | Novo Nordisk A/S | A stabilized pharmaceutical formulation comprising growth hormone and histidine |
US6204036B1 (en) * | 1995-03-09 | 2001-03-20 | Aventis Behring Gmbh | Stable transglutaminase preparations and processes for their production |
US6013773A (en) * | 1996-02-02 | 2000-01-11 | Mitsui Chemicals, Inc. | Pharmaceutical preparation containing human growth hormone |
WO1997039768A1 (en) * | 1996-04-24 | 1997-10-30 | Novo Nordisk A/S | A pharmaceutical formulation containing growth hormone, an amino acid and a non-ionic detergent |
WO2001049314A2 (en) * | 1999-12-30 | 2001-07-12 | Nps Allelix Corp. | Glp-2 formulations |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020012243A2 (en) | 2018-07-09 | 2020-01-16 | Abbott Laboratories Gmbh | Pharmaceutical composition containing a peptide |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6676589B2 (en) | Compstatin analogs with improved pharmacokinetic properties | |
EP1626983B1 (en) | Novel poly (ethylene glycol) modified erythropoietin agonists and uses thereof | |
ES2436152T3 (en) | Use of calcitonin in osteoarthritis | |
US8835389B2 (en) | Use of calcitonin for the treatment of RA | |
JP2003519195A (en) | GLP-2 preparation | |
JP2011184442A (en) | Il-21 derivatives | |
PT726778E (en) | ORAL ADMINISTRATION SYSTEM OF CHEMICALLY MODIFIED G-CSF PROTEINS | |
TW202317604A (en) | Hepcidin mimetics for treatment of hereditary hemochromatosis | |
KR20160012992A (en) | Compositions and methods of using islet neogenesis peptides and analogs thereof | |
JP2022535556A (en) | Parenteral Pharmaceutical Compositions of GLP1/2 Dual Agonists | |
EP3250225A1 (en) | Pharmaceutical composition for oral insulin administration comprising a tablet core and a polyvinyl alcohol coating | |
JP2025506374A (en) | Peptide inhibitors of interleukin-23 receptor and pharmaceutical compositions thereof | |
JP2022545265A (en) | Peptides for treating muscle atrophy | |
JP2007533298A (en) | Derivatives of IL-21 | |
WO2007033427A1 (en) | Stabilisation of peptides with basic amino acids | |
CN103096916B (en) | For the preparation of bG-CSF and variant thereof | |
WO2001000230A1 (en) | Drug composition containing lecithin-modified superoxide dismutase | |
JP2010533129A (en) | Site-specific PEGylated linear salmon calcitonin analogues | |
JP4880845B2 (en) | GRF-containing freeze-dried pharmaceutical composition | |
JP4931282B2 (en) | Bioactive peptide-containing powder | |
JPH07501543A (en) | blood regulating peptides | |
JP2009502779A (en) | Human growth hormone conjugated to a biocompatible polymer | |
WO2009127140A1 (en) | 7p peptide and its derivant, the use thereof | |
JP3459668B2 (en) | Pharmaceutical composition of stabilized calcitonin | |
TW201307381A (en) | Novel N-terminally modified insulin derivatives |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
DPE1 | Request for preliminary examination filed after expiration of 19th month from priority date (pct application filed from 20040101) | ||
NENP | Non-entry into the national phase |
Ref country code: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 06775012 Country of ref document: EP Kind code of ref document: A1 |