AU2003271745A1 - Beta-SHEET BREAKING PEPTIDES - Google Patents
Beta-SHEET BREAKING PEPTIDES Download PDFInfo
- Publication number
- AU2003271745A1 AU2003271745A1 AU2003271745A AU2003271745A AU2003271745A1 AU 2003271745 A1 AU2003271745 A1 AU 2003271745A1 AU 2003271745 A AU2003271745 A AU 2003271745A AU 2003271745 A AU2003271745 A AU 2003271745A AU 2003271745 A1 AU2003271745 A1 AU 2003271745A1
- Authority
- AU
- Australia
- Prior art keywords
- phe
- compound
- compound according
- amyloid
- disease
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title description 52
- 102000004196 processed proteins & peptides Human genes 0.000 title description 21
- 150000001875 compounds Chemical class 0.000 claims description 120
- 208000024827 Alzheimer disease Diseases 0.000 claims description 23
- -1 propionoyl Chemical group 0.000 claims description 17
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 16
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 16
- 201000010099 disease Diseases 0.000 claims description 14
- 239000003814 drug Substances 0.000 claims description 11
- 150000003839 salts Chemical class 0.000 claims description 11
- 206010059245 Angiopathy Diseases 0.000 claims description 9
- 201000004810 Vascular dementia Diseases 0.000 claims description 9
- 206010002022 amyloidosis Diseases 0.000 claims description 8
- 206010008111 Cerebral haemorrhage Diseases 0.000 claims description 7
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 claims description 6
- 208000017004 dementia pugilistica Diseases 0.000 claims description 6
- 206010019196 Head injury Diseases 0.000 claims description 5
- 230000002159 abnormal effect Effects 0.000 claims description 5
- 229910052739 hydrogen Inorganic materials 0.000 claims description 5
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 4
- 230000002265 prevention Effects 0.000 claims description 4
- 230000012846 protein folding Effects 0.000 claims description 4
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 229910052799 carbon Inorganic materials 0.000 claims 3
- 239000003085 diluting agent Substances 0.000 claims 1
- 210000004556 brain Anatomy 0.000 description 40
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 31
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 25
- 239000000243 solution Substances 0.000 description 20
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 14
- 230000008499 blood brain barrier function Effects 0.000 description 12
- 210000001218 blood-brain barrier Anatomy 0.000 description 12
- 230000000694 effects Effects 0.000 description 12
- 239000011347 resin Substances 0.000 description 12
- 229920005989 resin Polymers 0.000 description 12
- 238000002347 injection Methods 0.000 description 11
- 239000007924 injection Substances 0.000 description 11
- 238000000034 method Methods 0.000 description 11
- 102000004169 proteins and genes Human genes 0.000 description 11
- 108090000623 proteins and genes Proteins 0.000 description 11
- 210000002966 serum Anatomy 0.000 description 11
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 10
- 210000004369 blood Anatomy 0.000 description 10
- 239000008280 blood Substances 0.000 description 10
- 230000005764 inhibitory process Effects 0.000 description 10
- 239000000203 mixture Substances 0.000 description 10
- 238000010168 coupling process Methods 0.000 description 9
- 238000005859 coupling reaction Methods 0.000 description 9
- 208000005145 Cerebral amyloid angiopathy Diseases 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 8
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 8
- 230000003941 amyloidogenesis Effects 0.000 description 8
- 230000008878 coupling Effects 0.000 description 8
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 7
- 208000037259 Amyloid Plaque Diseases 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 7
- YZCKVEUIGOORGS-NJFSPNSNSA-N Tritium Chemical compound [3H] YZCKVEUIGOORGS-NJFSPNSNSA-N 0.000 description 7
- 229940024606 amino acid Drugs 0.000 description 7
- 150000001413 amino acids Chemical group 0.000 description 7
- 229910052722 tritium Inorganic materials 0.000 description 7
- 102000001049 Amyloid Human genes 0.000 description 6
- 108010094108 Amyloid Proteins 0.000 description 6
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 241000699670 Mus sp. Species 0.000 description 6
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 229940098773 bovine serum albumin Drugs 0.000 description 6
- JADVWWSKYZXRGX-UHFFFAOYSA-M thioflavine T Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C1=[N+](C)C2=CC=C(C)C=C2S1 JADVWWSKYZXRGX-UHFFFAOYSA-M 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 5
- 241000700159 Rattus Species 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- 230000002285 radioactive effect Effects 0.000 description 5
- 239000012730 sustained-release form 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
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-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
- 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 4
- 239000007821 HATU Substances 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 230000030833 cell death Effects 0.000 description 4
- 230000002490 cerebral effect Effects 0.000 description 4
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 4
- 238000000099 in vitro assay Methods 0.000 description 4
- 230000004941 influx Effects 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 238000013268 sustained release Methods 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 3
- 101710137189 Amyloid-beta A4 protein Proteins 0.000 description 3
- 102100022704 Amyloid-beta precursor protein Human genes 0.000 description 3
- 101710151993 Amyloid-beta precursor protein Proteins 0.000 description 3
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 239000007983 Tris buffer Substances 0.000 description 3
- 125000000217 alkyl group Chemical group 0.000 description 3
- DZHSAHHDTRWUTF-SIQRNXPUSA-N amyloid-beta polypeptide 42 Chemical compound C([C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O)[C@@H](C)CC)C(C)C)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C(C)C)C1=CC=CC=C1 DZHSAHHDTRWUTF-SIQRNXPUSA-N 0.000 description 3
- 238000013459 approach Methods 0.000 description 3
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 3
- 239000000969 carrier Substances 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 231100000433 cytotoxic Toxicity 0.000 description 3
- 230000001472 cytotoxic effect Effects 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 230000008021 deposition Effects 0.000 description 3
- 238000010511 deprotection reaction Methods 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 239000002552 dosage form Substances 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 210000002569 neuron Anatomy 0.000 description 3
- 230000001717 pathogenic effect Effects 0.000 description 3
- 230000035515 penetration Effects 0.000 description 3
- 238000010647 peptide synthesis reaction Methods 0.000 description 3
- 230000035699 permeability Effects 0.000 description 3
- 230000000144 pharmacologic effect Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 238000001356 surgical procedure Methods 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- 238000005533 tritiation Methods 0.000 description 3
- 239000003981 vehicle Substances 0.000 description 3
- WZZBNLYBHUDSHF-DHLKQENFSA-N 1-[(3s,4s)-4-[8-(2-chloro-4-pyrimidin-2-yloxyphenyl)-7-fluoro-2-methylimidazo[4,5-c]quinolin-1-yl]-3-fluoropiperidin-1-yl]-2-hydroxyethanone Chemical compound CC1=NC2=CN=C3C=C(F)C(C=4C(=CC(OC=5N=CC=CN=5)=CC=4)Cl)=CC3=C2N1[C@H]1CCN(C(=O)CO)C[C@@H]1F WZZBNLYBHUDSHF-DHLKQENFSA-N 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 206010012289 Dementia Diseases 0.000 description 2
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 2
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 239000007995 HEPES buffer Substances 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 2
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 108091006629 SLC13A2 Proteins 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000012317 TBTU Substances 0.000 description 2
- CLZISMQKJZCZDN-UHFFFAOYSA-N [benzotriazol-1-yloxy(dimethylamino)methylidene]-dimethylazanium Chemical compound C1=CC=C2N(OC(N(C)C)=[N+](C)C)N=NC2=C1 CLZISMQKJZCZDN-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 125000002252 acyl group Chemical group 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 239000000783 alginic acid Substances 0.000 description 2
- 235000010443 alginic acid Nutrition 0.000 description 2
- 229920000615 alginic acid Polymers 0.000 description 2
- 229960001126 alginic acid Drugs 0.000 description 2
- 150000004781 alginic acids Chemical class 0.000 description 2
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 229960005261 aspartic acid Drugs 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 235000011148 calcium chloride Nutrition 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 210000001715 carotid artery Anatomy 0.000 description 2
- 210000003710 cerebral cortex Anatomy 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000012377 drug delivery Methods 0.000 description 2
- 235000019439 ethyl acetate Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000010191 image analysis Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000007972 injectable composition Substances 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 2
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 2
- 210000004731 jugular vein Anatomy 0.000 description 2
- 239000007788 liquid Substances 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
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 2
- OSWPMRLSEDHDFF-UHFFFAOYSA-N methyl salicylate Chemical compound COC(=O)C1=CC=CC=C1O OSWPMRLSEDHDFF-UHFFFAOYSA-N 0.000 description 2
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 2
- 229960001412 pentobarbital Drugs 0.000 description 2
- 125000001147 pentyl group Chemical group C(CCCC)* 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 238000002953 preparative HPLC Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000017854 proteolysis Effects 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000031998 transcytosis Effects 0.000 description 2
- ZPGDWQNBZYOZTI-SFHVURJKSA-N (2s)-1-(9h-fluoren-9-ylmethoxycarbonyl)pyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1C(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 ZPGDWQNBZYOZTI-SFHVURJKSA-N 0.000 description 1
- 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
- CBPJQFCAFFNICX-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-methylpentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CC(C)C)C(O)=O)C3=CC=CC=C3C2=C1 CBPJQFCAFFNICX-IBGZPJMESA-N 0.000 description 1
- KSDTXRUIZMTBNV-INIZCTEOSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)butanedioic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CC(=O)O)C(O)=O)C3=CC=CC=C3C2=C1 KSDTXRUIZMTBNV-INIZCTEOSA-N 0.000 description 1
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 1
- TUSDEZXZIZRFGC-UHFFFAOYSA-N 1-O-galloyl-3,6-(R)-HHDP-beta-D-glucose Natural products OC1C(O2)COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC1C(O)C2OC(=O)C1=CC(O)=C(O)C(O)=C1 TUSDEZXZIZRFGC-UHFFFAOYSA-N 0.000 description 1
- PYHXGXCGESYPCW-UHFFFAOYSA-M 2,2-diphenylacetate Chemical compound C=1C=CC=CC=1C(C(=O)[O-])C1=CC=CC=C1 PYHXGXCGESYPCW-UHFFFAOYSA-M 0.000 description 1
- ZFFMLCVRJBZUDZ-UHFFFAOYSA-N 2,3-dimethylbutane Chemical group CC(C)C(C)C ZFFMLCVRJBZUDZ-UHFFFAOYSA-N 0.000 description 1
- OMGHIGVFLOPEHJ-UHFFFAOYSA-N 2,5-dihydro-1h-pyrrol-1-ium-2-carboxylate Chemical group OC(=O)C1NCC=C1 OMGHIGVFLOPEHJ-UHFFFAOYSA-N 0.000 description 1
- FBFNMTGUOBUGFQ-UHFFFAOYSA-M 2-(2,5-diphenyltetrazol-1-ium-1-yl)-4,5-dimethyl-1,3-thiazole;bromide Chemical compound [Br-].S1C(C)=C(C)N=C1[N+]1=C(C=2C=CC=CC=2)N=NN1C1=CC=CC=C1 FBFNMTGUOBUGFQ-UHFFFAOYSA-M 0.000 description 1
- LBLYYCQCTBFVLH-UHFFFAOYSA-M 2-methylbenzenesulfonate Chemical compound CC1=CC=CC=C1S([O-])(=O)=O LBLYYCQCTBFVLH-UHFFFAOYSA-M 0.000 description 1
- KLDLRDSRCMJKGM-UHFFFAOYSA-N 3-[chloro-(2-oxo-1,3-oxazolidin-3-yl)phosphoryl]-1,3-oxazolidin-2-one Chemical compound C1COC(=O)N1P(=O)(Cl)N1CCOC1=O KLDLRDSRCMJKGM-UHFFFAOYSA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- FZTIWOBQQYPTCJ-UHFFFAOYSA-N 4-[4-(4-carboxyphenyl)phenyl]benzoic acid Chemical compound C1=CC(C(=O)O)=CC=C1C1=CC=C(C=2C=CC(=CC=2)C(O)=O)C=C1 FZTIWOBQQYPTCJ-UHFFFAOYSA-N 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- XXSCONYSQQLHTH-UHFFFAOYSA-N 9h-fluoren-9-ylmethanol Chemical compound C1=CC=C2C(CO)C3=CC=CC=C3C2=C1 XXSCONYSQQLHTH-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 208000000044 Amnesia Diseases 0.000 description 1
- 102000009091 Amyloidogenic Proteins Human genes 0.000 description 1
- 108010048112 Amyloidogenic Proteins Proteins 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 241000416162 Astragalus gummifer Species 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- 102400000967 Bradykinin Human genes 0.000 description 1
- 101800004538 Bradykinin Proteins 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 1
- 201000000166 CST3-related cerebral amyloid angiopathy Diseases 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 206010051290 Central nervous system lesion Diseases 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 150000008574 D-amino acids Chemical class 0.000 description 1
- 208000031124 Dementia Alzheimer type Diseases 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 201000010374 Down Syndrome Diseases 0.000 description 1
- 239000001263 FEMA 3042 Substances 0.000 description 1
- 208000007487 Familial Cerebral Amyloid Angiopathy Diseases 0.000 description 1
- 239000001828 Gelatine Substances 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- AEMRFAOFKBGASW-UHFFFAOYSA-M Glycolate Chemical compound OCC([O-])=O AEMRFAOFKBGASW-UHFFFAOYSA-M 0.000 description 1
- QXZGBUJJYSLZLT-UHFFFAOYSA-N H-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-OH Natural products NC(N)=NCCCC(N)C(=O)N1CCCC1C(=O)N1C(C(=O)NCC(=O)NC(CC=2C=CC=CC=2)C(=O)NC(CO)C(=O)N2C(CCC2)C(=O)NC(CC=2C=CC=CC=2)C(=O)NC(CCCN=C(N)N)C(O)=O)CCC1 QXZGBUJJYSLZLT-UHFFFAOYSA-N 0.000 description 1
- 208000032849 Hereditary cerebral hemorrhage with amyloidosis Diseases 0.000 description 1
- 206010062767 Hypophysitis Diseases 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 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 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 244000246386 Mentha pulegium Species 0.000 description 1
- 235000016257 Mentha pulegium Nutrition 0.000 description 1
- 235000004357 Mentha x piperita Nutrition 0.000 description 1
- AFVFQIVMOAPDHO-UHFFFAOYSA-M Methanesulfonate Chemical compound CS([O-])(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-M 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 239000006057 Non-nutritive feed additive Substances 0.000 description 1
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- LRBQNJMCXXYXIU-PPKXGCFTSA-N Penta-digallate-beta-D-glucose Natural products OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 108010020346 Polyglutamic Acid Proteins 0.000 description 1
- 102000029797 Prion Human genes 0.000 description 1
- 108091000054 Prion Proteins 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- 102400000267 Rhomboid-related protein 2, N-terminal fragment Human genes 0.000 description 1
- 101800000645 Rhomboid-related protein 2, N-terminal fragment Proteins 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 238000003639 Student–Newman–Keuls (SNK) method Methods 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 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 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 229920001615 Tragacanth Polymers 0.000 description 1
- 102000004338 Transferrin Human genes 0.000 description 1
- 108090000901 Transferrin Proteins 0.000 description 1
- 208000037280 Trisomy Diseases 0.000 description 1
- 206010044688 Trisomy 21 Diseases 0.000 description 1
- 101800000716 Tumor necrosis factor, membrane form Proteins 0.000 description 1
- JPKKQJKQTPNWTR-KQAYXBCTSA-N [(1r,5s)-8-methyl-8-azabicyclo[3.2.1]octan-3-yl] (2r)-3-hydroxy-2-phenylpropanoate;sulfuric acid;hydrate Chemical compound O.OS(O)(=O)=O.C1([C@H](CO)C(=O)OC2C[C@H]3CC[C@@H](C2)N3C)=CC=CC=C1.C1([C@H](CO)C(=O)OC2C[C@H]3CC[C@@H](C2)N3C)=CC=CC=C1 JPKKQJKQTPNWTR-KQAYXBCTSA-N 0.000 description 1
- 235000011054 acetic acid Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000000274 adsorptive effect Effects 0.000 description 1
- 150000004703 alkoxides Chemical class 0.000 description 1
- VREFGVBLTWBCJP-UHFFFAOYSA-N alprazolam Chemical compound C12=CC(Cl)=CC=C2N2C(C)=NN=C2CN=C1C1=CC=CC=C1 VREFGVBLTWBCJP-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 150000001408 amides Chemical group 0.000 description 1
- KLOHDWPABZXLGI-YWUHCJSESA-M ampicillin sodium Chemical compound [Na+].C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C([O-])=O)(C)C)=CC=CC=C1 KLOHDWPABZXLGI-YWUHCJSESA-M 0.000 description 1
- 238000001949 anaesthesia Methods 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 210000002376 aorta thoracic Anatomy 0.000 description 1
- 101150031224 app gene Proteins 0.000 description 1
- 239000008135 aqueous vehicle Substances 0.000 description 1
- 229940072107 ascorbate Drugs 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 238000011888 autopsy Methods 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 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
- 230000002146 bilateral effect Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- QXZGBUJJYSLZLT-FDISYFBBSA-N bradykinin Chemical compound NC(=N)NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(=O)NCC(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CO)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)CCC1 QXZGBUJJYSLZLT-FDISYFBBSA-N 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 125000004744 butyloxycarbonyl group Chemical group 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- BPKIGYQJPYCAOW-FFJTTWKXSA-I calcium;potassium;disodium;(2s)-2-hydroxypropanoate;dichloride;dihydroxide;hydrate Chemical compound O.[OH-].[OH-].[Na+].[Na+].[Cl-].[Cl-].[K+].[Ca+2].C[C@H](O)C([O-])=O BPKIGYQJPYCAOW-FFJTTWKXSA-I 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 150000001718 carbodiimides Chemical class 0.000 description 1
- 150000007942 carboxylates Chemical class 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000000423 cell based assay Methods 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229940114081 cinnamate Drugs 0.000 description 1
- 229940001468 citrate Drugs 0.000 description 1
- 230000001149 cognitive effect Effects 0.000 description 1
- 229940075614 colloidal silicon dioxide Drugs 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 229940125904 compound 1 Drugs 0.000 description 1
- 229940125773 compound 10 Drugs 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 239000002577 cryoprotective agent Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229960004132 diethyl ether Drugs 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 1
- BFMYDTVEBKDAKJ-UHFFFAOYSA-L disodium;(2',7'-dibromo-3',6'-dioxido-3-oxospiro[2-benzofuran-1,9'-xanthene]-4'-yl)mercury;hydrate Chemical compound O.[Na+].[Na+].O1C(=O)C2=CC=CC=C2C21C1=CC(Br)=C([O-])C([Hg])=C1OC1=C2C=C(Br)C([O-])=C1 BFMYDTVEBKDAKJ-UHFFFAOYSA-L 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000035557 fibrillogenesis Effects 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 229940050411 fumarate Drugs 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 235000011087 fumaric acid Nutrition 0.000 description 1
- LRBQNJMCXXYXIU-QWKBTXIPSA-N gallotannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@H]2[C@@H]([C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-QWKBTXIPSA-N 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 244000144993 groups of animals Species 0.000 description 1
- 229940093915 gynecological organic acid Drugs 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 125000004051 hexyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 235000001050 hortel pimenta Nutrition 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 210000004283 incisor Anatomy 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 238000007913 intrathecal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- ZLVXBBHTMQJRSX-VMGNSXQWSA-N jdtic Chemical compound C1([C@]2(C)CCN(C[C@@H]2C)C[C@H](C(C)C)NC(=O)[C@@H]2NCC3=CC(O)=CC=C3C2)=CC=CC(O)=C1 ZLVXBBHTMQJRSX-VMGNSXQWSA-N 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 210000005240 left ventricle Anatomy 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 229960003136 leucine Drugs 0.000 description 1
- 239000006193 liquid solution Substances 0.000 description 1
- 239000006194 liquid suspension Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 229940049920 malate Drugs 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- IWYDHOAUDWTVEP-UHFFFAOYSA-M mandelate Chemical compound [O-]C(=O)C(O)C1=CC=CC=C1 IWYDHOAUDWTVEP-UHFFFAOYSA-M 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- STZCRXQWRGQSJD-GEEYTBSJSA-M methyl orange Chemical compound [Na+].C1=CC(N(C)C)=CC=C1\N=N\C1=CC=C(S([O-])(=O)=O)C=C1 STZCRXQWRGQSJD-GEEYTBSJSA-M 0.000 description 1
- 229940012189 methyl orange Drugs 0.000 description 1
- 229960001047 methyl salicylate Drugs 0.000 description 1
- 125000000250 methylamino group Chemical group [H]N(*)C([H])([H])[H] 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- YZMHQCWXYHARLS-UHFFFAOYSA-N naphthalene-1,2-disulfonic acid Chemical compound C1=CC=CC2=C(S(O)(=O)=O)C(S(=O)(=O)O)=CC=C21 YZMHQCWXYHARLS-UHFFFAOYSA-N 0.000 description 1
- PSZYNBSKGUBXEH-UHFFFAOYSA-N naphthalene-1-sulfonic acid Chemical compound C1=CC=C2C(S(=O)(=O)O)=CC=CC2=C1 PSZYNBSKGUBXEH-UHFFFAOYSA-N 0.000 description 1
- 210000002682 neurofibrillary tangle Anatomy 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 125000004433 nitrogen atom Chemical group N* 0.000 description 1
- 239000002687 nonaqueous vehicle Substances 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- WLJNZVDCPSBLRP-UHFFFAOYSA-N pamoic acid Chemical compound C1=CC=C2C(CC=3C4=CC=CC=C4C=C(C=3O)C(=O)O)=C(O)C(C(O)=O)=CC2=C1 WLJNZVDCPSBLRP-UHFFFAOYSA-N 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 229940124531 pharmaceutical excipient Drugs 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- RLOWWWKZYUNIDI-UHFFFAOYSA-N phosphinic chloride Chemical compound ClP=O RLOWWWKZYUNIDI-UHFFFAOYSA-N 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 230000001817 pituitary effect Effects 0.000 description 1
- 239000010318 polygalacturonic acid Substances 0.000 description 1
- 229920002643 polyglutamic acid Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000007425 progressive decline Effects 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 229960002429 proline Drugs 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 238000011552 rat model Methods 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 230000028527 righting reflex Effects 0.000 description 1
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 1
- 229940081974 saccharin Drugs 0.000 description 1
- 235000019204 saccharin Nutrition 0.000 description 1
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 description 1
- 210000003625 skull Anatomy 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000008247 solid mixture Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 235000015523 tannic acid Nutrition 0.000 description 1
- 229940033123 tannic acid Drugs 0.000 description 1
- 229920002258 tannic acid Polymers 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 229940095064 tartrate Drugs 0.000 description 1
- 125000004213 tert-butoxy group Chemical group [H]C([H])([H])C(O*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M trans-cinnamate Chemical compound [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- 239000012581 transferrin Substances 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 238000007492 two-way ANOVA Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 210000005166 vasculature Anatomy 0.000 description 1
- 230000002227 vasoactive effect Effects 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4711—Alzheimer's disease; Amyloid plaque core protein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Neurology (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Veterinary Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Animal Behavior & Ethology (AREA)
- Neurosurgery (AREA)
- Public Health (AREA)
- Psychiatry (AREA)
- Biochemistry (AREA)
- Hospice & Palliative Care (AREA)
- Urology & Nephrology (AREA)
- Cardiology (AREA)
- Heart & Thoracic Surgery (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Vascular Medicine (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Description
WO 2004/005336 PCT/EP2003/050287 1 |-Sheet breaking peptides Field of Invention The invention relates to the field of 3-sheet breaking peptides, particularly their use in the treatment of diseases such as Alzheimer's disease, Dementia pugilistica (including head 5 trauma), Hereditary Cerebral Haemorrhage with amyloidosis of the Dutch type (HCHWA D) and vascular dementia with amyloid angiopathy. Background of the Invention Alzheimer's disease (AD), first described by the Bavarian psychiatrist Alois Alzheimer in 1907, is a progressive neurological disorder that begins with short-term memory loss and is 10 characterized by a progressive decline in cognitive fnimction and behaviour. Progression of the disease leads to disorientation, impairment of judgment, reasoning, attention and speech and, ultimately, dementia. The course of the disease usually leads to death in a severely debilitated, immobile state between four and 12 years after onset. AD has been estimated to afflict 5 to 11 percent of the population over age 65 and as much as 47 percent of the 15 population over age 85. The societal cost for managing AD is very high, primarily due to the extensive custodial care required for AD patients. Despite continuous efforts aimed at understanding the physiopathology of AD, there is currently no treatment that significantly retards the progression of the disease. Pathologically, AD is characterized by the presence of distinctive lesions in the victim's 20 brain, revealed on autopsy. These brain lesions include abnormal intracellular filaments called neurofibrillary tangles (NTFs) and extracellular deposits of amyloidogenic proteins in senile, or amyloid, plaques. Amyloid deposits are also present in the walls of cerebral blood vessels of AD patients. The major protein constituent of amyloid plaques has been identified as a 4.3 kiloDalton peptide called 3-amyloid peptide (AP3) 1 . Diffuse deposits of AP3 are 25 fr-equently observed in normal adult brains, whereas AD brain tissue is characterized by more compacted, dense-core 1-amyloid plaques.
2 These observations suggest that A3 deposition precedes, and contributes to, the destruction of neurons that occurs in AD 3 . In further support of a direct pathogenic role for AP3, P3-amyloid has been shown to be toxic to mature neurons, both in culture and in vivo 4
.
WO 2004/005336 PCT/EP2003/050287 2 Patients with hereditary cerebral haemorrhage with amyloidosis-Dutch-type (HCHWA-D), which is characterized by diffuse P-amyloid deposits within the cerebral cortex and cerebrovasculature, have been shown to have a point mutation that leads to an amino acid substitution within AD.
5 5 AP3 has also been implicated in vascular dementia with amyloid angiopathy 6 and dementia pugilistica.
7 Natural AP3 is derived by proteolysis from a much larger protein called the amyloid precursor protein (APP) 8 . The APP gene maps to chromosome 21, thereby providing an explanation for the P-amyloid deposition seen at an early age in individuals with Down's syndrome, 10 which is caused by trisomy of chromosome 219. Naturally-occurring AP3, derived from proteolysis of APP, is 39 to 43 amino acid residues in length, depending on the carboxyl-terminal end point, which exhibits heterogeneity. The predominant circulating form of A3 in the blood and cerebrospinal fluid of both AD patients and normal adults is API31-40 10 . However, AP142 and AP3143 are also found in 3-amyloid 11 15 plaques Considerable evidence has accumulated that the pathogenicity of AP results from a change in protein conformation 1 2 . It is believed that a critical event leading to pathology in Alzheimer's disease, Vascular Dementia with amyloid angiopathy and HCHWA-D is the refolding of a natural and non-pathogenic protein, to yield a pathogenic form. The refolding 20 alters the secondary and tertiary structure of the protein without changing its primary structure. Amyloid is a generic term that is applied to fibrillar aggregates that have a common structural motif: a [3-pleated sheet conformation' 3 . These aggregates exhibit special tinctorial properties, including the ability to emit a green birefringent glow after staining with Congo 25 red, and the capacity to bind the fluorochrome Thioflavin S 14 . These tinctorial properties form the basis of assays used to detect 3-amnyloid deposits. One approach to the treatment and prevention of Alzheimer's disease has been to develop short peptides having some sequence homology to the natural protein sequence believed to WO 2004/005336 PCT/EP2003/050287 3 be involved in amyloid formation, but also having one or more amino acids that disfavour or destabilise the formation of P-pleated sheet conformations1 5 . The peptides prevent the aggregation of -amyloid, and thereby prevent its cytotoxic effects. This approach has been suggested in Alzheimer's disease and in prion-related disorders 16
,
17 and has lead to the 5 P3-sheet breaking peptides shown below, amongst others: 1 1 0 1- 0 OH 00 '0 0 OH HN J M N N ,'AJOH N N NH, H H H H 0 O " 0 O0 0 WO 96/39834 (New York Univeristy) WO 01/34631 (Axonyx, Inc.) US 6,319,498 (Praecis Pharmaceuticals) proposes -sheet breaking peptides based on A3, and exemplifies amino-terminally-biotinylated peptides. US 6,303,567 (Praecis Pharmaceuticals) proposes peptides based on the -amyloid peptide, but consisting entirely 10 of D-amino acids, as -sheet breaking peptides. Others have proposed for preventing -strand association, peptides comprising a -strand forming section of peptide which forms a P-strand and associate as such with a target -strand, and wherein the -strand forming section of peptide comprises a sequence of at least four consecutive a-L-amino acid, all of which sterically permit the P-stand forming 15 section of peptide to form a -stand, and at least one of which is a Nu-substituted a-L amino-acid residue, and any two successive Nec-substituted a-L-amino-acid residues are separated by an odd number of consecutive Na-substituted ea-L-amino-acid residues. 18 Alternatively, peptides containing short 1-strands and N-methyl amino acids at alternate positions with or without N-a-acetylated amino-acids have also been developed for 20 inhibition of fibril formnation.19 While the known -sheet breaking peptides have provided valuable information and may have utility in treating Alzheimer's disease, the development of peptide drugs is severely limited by the fact that natural peptides are subject to degradation and rapid metabolism in 25 the gut, the liver and in circulation. Furthermore, the desired site of action for treatment of many amyloid-related disorders is in the brain, and peptides, like many other molecules, may WO 2004/005336 PCT/EP2003/050287 4 have difficulty penetrating the blood brain barrier. The brain itself is also replete with peptidases, which degrade peptide molecules. Summary of the invention 5 It is an object of the invention to provide a P3-sheet breaking peptide with improved pharmacological profile. In a first aspect, the invention provides a compound of the general Formula I: 0 0 0 OH H H R NR N NR R 6 NR H ONR O H -N H, 000 (I) wherein:
R
1 is selected from H and optionally substituted C 2
-C
6 -acyl, preferably acetyl; 10 R, R 3 , R 4 and R 5 are independently selected from H and optionally substituted C 1
-C
6 -alkyl and wherein at least one among R 2 , R 3 , R 4 and R 5 is optionally substituted C 1
-C
6 -alkyl, preferably methyl;
R
6 is selected from OH and NR 7
R
8 , wherein R 7 and R 8 are independently H and optionally substituted C 1
-C
6 -alkyl, preferably NH 2 ; and salts and tritiated derivatives thereof. 15 In a second aspect, the invention provides a compound of Formula I for use as a medicament. In a third aspect, the invention provides a pharmaceutical composition comprising a 20 compound of Formula I, together with a pharmaceutically acceptable excipient or carrier. In a fourth aspect, the invention provides a use of a compound of Formula I for the preparation of a medicament for the treatment or prevention of a disease or condition selected from Alzheimer's disease, Dementia pugilistica (including head trauma), Hereditary WO 2004/005336 PCT/EP2003/050287 5 Cerebral Haemorrhage with amyloidosis of the Dutch type (HCHWA-D) and vascular dementia with amyloid angiopathy. In a fifth aspect, the invention provides a use of a compound of Formula I for the treatment 5 or prevention of a disease or condition selected from Alzheimer's disease, Dementia pugilistica (including head trauma), Hereditary Cerebral Haemorrhage with amyloidosis of the Dutch type (HCHWA-D) and vascular dementia with amyloid angiopathy. In a sixth aspect, the invention provides a method of treating Alzheimer's disease, Dementia 10 pugilistica (including head trauma), Hereditary Cerebral Haemorrhage with amyloidosis of the Dutch type (HICHWA-D) and vascular dementia with amyloid angiopathy, comprising administering to a patient in need thereof an effective amount of a compound of Formula I. In a seventh aspect, the invention provides a use of a compound of Formula I for the 15 preparation of a medicament for the treatment of a disease associated with abnormal protein folding into amyloid and amyloid-like deposits. In an eighth aspect, the invention provides a use of a compound of Formula I for the treatment of a disease associated with abnormal protein folding into amyloid and 20 amyloid-like deposits. In a ninth aspect, the invention provides a method of treating a disease associated with abnormal protein folding into amyloid and amyloid-like deposits, comprising administering to a patient in need thereof an effective amount of a compound of Formula I. 25 Brief description of the drawings Figure 1 shows the in vitro activity of compounds of the invention using a fibrillogenesis assay. The compounds were screened for activity using an assay based on quantitating amyloid fibril formation of synthetic AP31-42. AP 3 1-42 (110 pM) in 0.1 M Tris, pH 7.4 was 30 incubated alone or in the presence of a ten-fold molar excess of each of the compounds during 5 days at 37 0 C. Thereafter, the amount of amyloid fibrils was measured using the Thioflavin T fluorescent method, as described in the Examples. Values correspond to the WO 2004/005336 PCT/EP2003/050287 6 percentage of inhibition of amyloid formation and are expressed relative to the activity of the comparative compound of Example 8 (100%). Figure 2 shows a graph representing the calculated % of injected dose of compound of 5 Example 7 taken up per g of brain vs. time. Mice are injected with 0.2 ml of lactate Ringer's solution containing 1% BSA and tritium labelled peptide ("hot") of Example 7 (100 000 cpm/ml of tritium radioactive peptide). Figure 3A shows the in vivo activity of the compound of Example 1 and the comparative 10 compound of Example 8 in a rat model of cerebral amyloidosis. A representative image shows the difference in amyloid size between the groups of animals treated with vehicle, the comparative compound of Example 8 and the compound of Example 1. Figure 3B shows a graph depicting the amyloid area in the different animals as estimated by 15 image analysis after immunohistochemistry. Detailed description of the invention The compounds of the invention are P3-sheet breaking peptides with improved pharmacological profile over known -sheet breaking peptides. 20 P-sheet breaking activity can be detected using, for example, an in vitro assay, such as that described by Soto et al.
14 , which measures the ability of test compounds to prevent amyloid fibril formation. 20 25 Amyloid fibrils are cytotoxic, inducing cell death by apoptosis . Compounds of the invention can be tested for their ability to prevent cell death induced by amyloid fibrils. Results are reported in the Examples. A compound having an improved pharmacological profile is considered to be a compound 30 having an increased in vitro activity, as measured by either or both of the in vitro assays described herein, an increased stability in plasma and/or in brain homogenate, or an increased ability to prevent amyloid deposition in vivo in rat brain, as compared with known WO 2004/005336 PCT/EP2003/050287 7 compounds. "Improved" encompasses those compounds showing an increase in any one of these parameters, or in more than one. Preferably the improvement will be a statistically significant one, preferably with a probability value of < 0.05. Methods of determining the statistical significance of results are well known and documents in the art, and any 5 appropriate method may be used. In a preferred group of compounds of Formula I, R 1 is selected from formyl, acetyl, propionoyl and butyroyl. Particularly preferably R' is acetyl. 10 In another preferred group of compounds of Formula I, R 6 is NHMe or NH 2 , particularly preferably NI-I 2 . A particular embodiment of the invention includes compounds of Formula I wherein R 2 , R ,
R
4 and R 5 are selected from H, methyl and ethyl, particularly preferably H and methyl and at 15 least one among R 2 , R 3 , R 4 and R 5 is selected from methyl and ethyl, preferably methyl. In a particularly preferred group of compounds of Formula I, R 3 is methyl, R 2 is H and R 4 and R 5 are selected from H, methyl. 20 In another particularly preferred group of compounds, R 3 is methyl and R 2 , R 4 and R 5 are H. In another particularly preferred embodiment, the stereochemistry at all the cx-carbon atoms is L; 25 The compounds of the invention may be isolated and purified as salts. Such salts fall within the scope of the invention. For the purposes of administration to a patient, it is desirable that the salts be pharmaceutically acceptable.
"C
1
-C
6 -alkyl" refers to monovalent branched or unbranched alkyl groups having 1 to 6 carbon atoms. This term is exemplified by groups such as methyl, ethyl, n-propyl, isopropyl, 30 n-butyl, isobutyl, tert-butyl, pentyl, hexyl and the like.
WO 2004/005336 PCT/EP2003/050287 8
"C
1 -Cs-alkyl" refers to monovalent branched or unbranched alkyl groups having 1 to 5 carbon atoms. This term is exemplified by groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl and the like. 5 "C 2
-C
6 Acyl" refers to a group -C(O)R where R includes "Ci-Cs-alkyl" groups. This term is exemplified by groups such as formyl, acetyl, propionoyl and butyroyl. "Pharmaceutically acceptable salts" refers to salts of the compounds of Formula I that retain the desired biological activity. Examples of such salts include, but are not restricted to, acid 10 addition salts formed with inorganic acids (e.g. hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, fumaric acid, maleic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalene sulfonic acid, naphthalene disulfonic acid, and polygalacturonic acid. Said 15 compounds can also be administered as pharmaceutically acceptable quaternary salts known by a person skilled in the art, which specifically include the quaternary ammonium salts of the formula -NR, R', R" + Z, wherein R, R', R" is independently hydrogen, alkyl, or benzyl, and Z is a counter ion, including chloride, bromide, iodide, alkoxide, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, 20 glycolate, maleate, malate, fumarate, citrate, tartrate, ascorbate, cinnamate, mandelate, and diphenylacetate). When employed as pharmaceuticals, the compounds of the invention are typically administered in the form of a pharmaceutical composition. Such compositions can be 25 prepared in a manner well known in the pharmaceutical art and comprise at least one active compound. Generally, the compounds of the invention are administered in a pharmaceutically effective amount. The amount of the compound actually administered will typically be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound 30 administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.
WO 2004/005336 PCT/EP2003/050287 9 The pharmaceutical compositions of the invention can be administered by a variety of routes including oral, rectal, transdermal, intrathecal, subcutaneous, intravenous, intramuscular, and intranasal. Preferably the compounds of the invention are administered by subcutaneous, intramuscular or intravenous injection or infusion. 5 In a preferred embodiment of the invention, a compound of the invention is fused to a carrier molecule, a peptide or a protein that promotes the crossing of the blood brain barrier ("BBB"). This serves for proper targeting of the molecule to the site of action in those cases, in which the CNS is involved in the disease. Modalities for drug delivery through the BBB 10 entail disruption of the BBB, either by osmotic means or biochemically by the use of vasoactive substances such as bradykinin. Other strategies to go through the BBB may entail the use of passive diffusion and the use of endogenous transport systems, including carrier-mediated transporters such as glucose and amino acid carriers; receptor-mediated transcytosis for insulin or transferrin; adsorptive-mediated transcytosis. Strategies for drug 15 delivery behind the BBB further include intracerebral implantation. Depending on the intended route of delivery, the compounds may be formulated as injectable or oral compositions. The compositions for oral administration can take the form of bulk liquid solutions or suspensions, or bulk powders. More commonly, however, the 20 compositions are presented in unit dosage forms to facilitate accurate dosing. The term "unit dosage forms" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a pre-determined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include pre-filled, pre-measured 25 ampoules or syringes of the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions. In such compositions, the compound of the invention is usually a minor component (from about 0.1 to about 50% by weight or preferably from about 1 to about 40% by weight) with the remainder being various vehicles or carriers and processing aids helpful for forming the desired dosing form. 30 Liquid forms suitable for oral administration may include a suitable aqueous or non-aqueous vehicle with buffers, suspending and dispensing agents, colorants, flavours and the like. Solid forms may include, for example, any of the following ingredients, or compounds of a WO 2004/005336 PCT/EP2003/050287 10 similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatine; an excipient such as starch or lactose; a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavouring agent such as 5 peppermint, methyl salicylate, or orange flavouring. Injectable compositions are typically based upon injectable sterile saline or phosphate buffered saline or other injectable carriers known in the art. The above-described components for orally administered or injectable compositions are 10 merely representative. Further materials as well as processing techniques and the like are known to the skilled practitioner.
21 The compounds of this invention can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials is also known to the skilled practitioner.
22 2 3
'
24 15 The compounds of the invention prevent the aggregation of A3 associated with the onset and progression of Alzheimer's disease, Dementia pugilistica (including head trauma), Hereditary Cerebral Haemorrhage with amyloidosis of the Dutch type (HCHWA-D) and vascular dementia with amyloid angiopathy. In a preferred method of use of the compounds, 20 administration of the compounds is by injection or infusion, at periodic intervals. The administration of a compound of the invention should preferably begin before any symptoms are detected in the patient, and should continue thereafter. Patients at a high risk for developing Alzheimer's disease, Hereditary Cerebral Haemorrhage with amyloidosis of the Dutch type (HCHWA-D) and vascular dementia with amyloid angiopathy include those with 25 a familial history of these diseases. Still a further aspect of the present invention is a process for preparing the compounds of Formula I. The compounds of the invention may be prepared from readily available starting materials using the following general methods and procedures. It will be appreciated that where typical or preferred experimental conditions (i.e., reaction temperatures, time, moles 30 of reagents, solvents, etc.) are given, other experimental conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimisation procedures.
WO 2004/005336 PCT/EP2003/050287 11 The compounds of the invention may be prepared using methods of peptide synthesis known to the skilled practitioner.25 In a preferred embodiment, the compounds of the invention are synthesised using solid-phase methods. 5 A preferred route to the compounds of the invention is depicted in Scheme 1, and particular examples are given in the Examples that follow. Abbreviations: 10 The following abbreviations are hereinafter used in the accompanying examples: min (minute), hr (hour), g (gram), mg (milligram), mmol (millimole), eq (equivalent), ml (milliliter), ptl (microliter), mm (millimeter), nm (nanometer), pm (micrometer), A (Angstrim), cpm (counts per minute), Ci (Curies), i. v. (intra veinous), ACN (acetonitrile), BBB (Blood Brain Barrier), BOP (benzotriazol-1-yl-oxcy-tris-(dimethylamino) 15 phosphonium hexafluorophosphate), BOP-Cl (bis(2-oxo-3-oxazoldinyl) phosphinic chloride), Boc (butoxycarbonyl), BSA (Bovine Serum Albumin), Cbz (carboxybenzyl), DCM (dichloromethane), DIC (diisopropyl . carbodiimide), DCC (dicyclohexylcarbodiimide), DIEA (diisopropyl ethylamine), DMAP (4-dimethylamino pyridine), DMF (dimethylformamide), DMSO (Dimethyl Sulfoxide), EDC (1-(3 20 dimethylaminopropyl)-3-ethlyl-carbodiimide hydro-chloride), EtOAc (ethyl acetate), Et20 (diethylether), Fmoc (9-fluorenylmethoxycarbonyl), HATU (0-(7-azabenzotriazol-1-yl) 1,1,3,3-tetramethyluronium hexafluorophosphate), HEPES (N-2-hydroxyethylpiperazine-N' 2-ethanesulfonic acid), HOBt (1-hydroxybenzotriazole), rt (room temperature), MTT (3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), PBS (Phosphate Buffered 25 Saline), PyBOP (benzotriazol- 1 -yloxy)tripyrrolidino-phosphonium hexafluoro-phosphate), PyBrop (bromo-tris-pyrrolidino-phosphonium hexafluorophosphate), TBTU (O benzotriazolyl-N,N,N',N'-tetramethyl-uronium-tetrafluoro-borate), TEA (triethylamine), TFA (trifluoro-acetic acid), THF (tetrahydrofuiran), ThT (Thioflavine T). N(Me), in the peptide sequence stands for a methyl group branched on the N atom of the 30 amino acid.
WO 2004/005336 PCT/EP2003/050287 12 Synthesis of compounds of the invention: The compounds according to formula I may be prepared from readily available starting materials. Examples of synthetic pathways will be described below. 5 General protocol: A preferred pathway for preparing pentapeptides according to the general Formula I, wherein R 2 , R', R 4 and R 5 are defined above is described in Scheme 1. Scheme 1: Fmoc-N Fmoc-AAx-OH Fmoc-N b) (11) coupling cycle 0 number: n 0 piperidine 20% in DMF 1 fmocs / OH N- a) NRO N NR 4 0 = Fmoc-Rink-Amide resin H-N 2 fmoc OH Fmoc-AA x -OH 3 fmoc / OH HATU, DIEA fm4 N 4 OR fmoc 0 n N o OH 5 fmoc 'OH /21 NR 0 1 0 1-0 In a preferred embodiment the peptides of Formula (I) may be synthesized on a solid support using, for example, preferred Rink-Amide resin. Typically the resin is washed with DCM, DMF, THF and again DCM three times.
WO 2004/005336 PCT/EP2003/050287 13 a) Resin deprotection step: A solution of piperidine in DMF (20-50%) is applied to remove the Fmoc-protecting group from the resin. The resin is gently shacked for 30 minutes to 1lhour in such a solution. After draining, the resin is washed with DCM, DMF, THF and again DCM each three times. 5 b) Coupling step: A solution of Fmoc-aspartic acid (f3-O-t-butyl) and the resin are shaken with 1.5 equivalents of coupling reagent, such as PyBOP, TBTU, HATU, BOP-C1, PyBrop, BOP, EDC, DIC, DCC, preferably HATU, and 3 equivalents of a tertiary base, such as NMP, triethylamine, diisopropyl-ethylamine or any tertiary base with similar pKa, preferably 10 diisopropylethylamine. After 2 to 15 hours at between 25 to 40 0 C, the resin is drained and washed with DCM, DMF, THF and again DCM three times. For the remaining four amino acids the deprotection and coupling steps (Scheme I, coupling cycles 2 to 5) are applied sequentially in the same manner as described above, using the appropriate Fmoc-protected amino acid of formula (II) to yield resin-bound compound of 15 Formula (III). After the fifth Fmoc-amino acid-coupling step, the group R 1 is introduced as follows. c) Introduction of R': The Fmoc moiety is removed from compound of Formula (III) as described above in the deprotection step and the group R' is introduced. 20 For compounds of formula I in which R 1 is an unsubstituted acyl group (scheme 2), a freshly prepared solution of R'COC1 in pyridine (20 equivalents) is shaken with the resin-bound compound (compound of formula III) for two to 20 hours at 25 to 40 0 C (scheme 2). The resin is extensively washed with DCM, DMF, THF and again DCM three times before being submitted to the cleavage step. The resulting compound wherein the R 1 moiety is present is 25 described by Formula (IV).
WO 2004/005336 PCT/EP2003/050287 14 Scheme 2: 10
-
0 0N 0 NRi H I) fmoc.2NR N N N( fmcN NR / NR 0 NR 3 0 - 0 i.:piperidine 20% in DMF [ ii.: RICOCI, pyridine c) 4 o O O 0
NR
2 N 0 3 N (IV) O NR O NR0O TFA, 95% in DCM d) D, 0 O 40 OH 2 k NR R 6 N ON NR N R 0O\ 0 0 d) Cleavage step: A freshly prepared solution of 95% TFA in DCM is shaken with the resin for one to four 5 hours at room temperature. This cleaves the compound from the resin as a C-terminal amide and removes the TFA-sensitive protecting groups. The solution is collected and the treatment is repeated two times. The solutions are collected and evaporated to dryness. The crude compound of Formula (I) is purified by preparative HPLC using conditions as described below. 10 Tritiation protocol: Tritiated derivatives of compound of Formula I can be prepared following the general protocol coupling a 3,4-dehydro-proline residue in step 4. The Dehydro-Pro peptide (2 mg) is mixed with 10% palladium on calcium carbonate (10 15 mg) and DMF (1 ml) in a tritiation vessel. The mixture is stirred under tritium gas (5 Ci) for WO 2004/005336 PCT/EP2003/050287 15 3 hours. The solution is filtered and labile tritium removed by repeated evaporations to dryness with ethanol. Crude reaction mixture was purified by high performance liquid chromatography in the following system: On a Vydac C18 Protein and Peptide column (250 x 9.6 mm) using a 5 water: acetonitrile: TFA gradient system. The detection is performed by a radioactive detector and a UV detector (220 nm). Product collected, evaporated to dryness, re-dissolved in dispensing eluent. The other purifications are performed as followed: Preparative HPLC Waters Prep LC 4000 System equipped with columns Prep Nova-Pak® HR C186 ptm 60A, 40x30mm (up to 100 10 mg) or 40x300 mm (up to 1g). All the purifications were performed with a gradient of MeCN/H 2 0 0.09% TFA. The following building blocks are commercially available from Bachem, Switzerland, Fmoc-L-phenylalanine, Fmoc-(P3-OtBu)-L-aspartic acid, Fmoc-L-leucine, Fmoc-L-proline, Fmoc-N-Me-L-phenylalanine, Fmoc-N-Me-(OtBu)-L-aspartic acid, Fmoc-N-Me-L- leucine. 15 Coupling reagents are commercially available from Novabiochem, Switzerland. Generally, the peptide derivatives according to the general Formula (I) can be synthesized using standard peptide synthesis techniques either in solution or on solid phase. In both approaches typical coupling reagents are used, which are known to the person skilled in the art. 20 It will be also appreciated that where typical or preferred experimental conditions (i.e. reaction temperatures, time, moles of reagents, solvents etc.) are given, other experimental conditions can also be used unless otherwise stated. Optimum reaction conditions may vaiy with the particular reactants or solvents used, but such conditions will be appreciated by the person skilled in the art. 25 Examples The compounds of Examples 1 to 7 are preferred embodiments of the invention. Structures of compounds of Examples 1 to 6 are presented in Table 1 below. 30 Compounds of Examples 1, 2, 3, 4, 5 and 6 may be synthesized according to protocol presented in scheme 1.
WO 2004/005336 PCT/EP2003/050287 16 Compound of Example 7 stands for tritiated compound of Example 1 which may be synthesized according to protocol presented in scheme 1 using a 3,4-dehydro-proline amino acid instead of Proline in step 4. The peptide is then tritiated according to the tritiation 5 protocol above. Compound of Example 7 had the specific activity of 42 Ci/mmol. Table 1 Example Structure 1 o 0 N HO N N NH O H N 0 0 0 HN 0 HO N NH N N N 0 H I H 0 O O H 2 N 3 OH N N NH, 0 O H OO 'O 4 OH 0 H N 0 0 N WO 2004/005336 PCT/EP2003/050287 17 Example Structure o 5 o OH H o O'O N N N N N 0 H 0 0 The mass of compounds of Examples 1 to 7 are presented in Table 2 below. Table 2 Example Theoretical Experimental No Mass Mass 1 692.8 692.3 2 692.8 692.3 3 706.8 706.8 4 706.8 707.4 WO 2004/005336 PCT/EP2003/050287 18 Example Theoretical Experimental No Mass Mass 5 706.8 706.5 6 720.9 720.4 7 690.8 691.5 Comparative Example 8 The following compound is disclosed in WO 01/34631 (Axonyx Inc.), and is included as a 5 reference compound. Example 8: 0 0 0 0 OH O O OH N N N NH 2 H /N- N H OH O O . - 0 10 Example 9 : Biological assays In vitro assays ofpeptide stability. The stability of the compounds of the invention can be assayed in comparison with the reference compound (Example 8). 15 WO 2004/005336 PCT/EP2003/050287 19 Peptides were prepared as a l gg/l solution in water. 20 Ll of the peptide solution was diluted in 80 gl of fresh human plasma or 10% rat brain homogenate (prepared in PBS). The resulting solution was incubated at 37 0 C for different times and the reaction was stopped by adding a complete cocktail of protease inhibitors. The bulk of plasma and brain proteins (but 5 none of the peptide) were precipitated in cold methanol (mix/MeOH, 4/5, v/v) during one hour at -20 0 C. The precipitated proteins were pelleted by centrifugation (10000g, 10 min, 4oC). The supernatant, containing the peptide, was concentrated 5 times under vacuum and separated by reverse-phase HPLC. The area of the peak corresponding to the intact peptide was measured and compared with an equivalent sample incubated in PBS. The results of are 10 listed in Table 3 as "t/ human plasma" and "t/ rat brain homogenate". The values compare favourably with those obtained for the reference compound of Example 8. Table 3. In vitro half-lives of various peptides Example no t/, human plasma t 2 rat brain homogenate 1 >24h >24h 4 >24h 5h 5 >24h >24h 6 >24h >24h 8 >24h 15 min 15 In vitro assays of activity. The activity of the compound of the invention in inhibiting the formation of aggregated fibrils can be tested by following the changes in fluorescence signal of a fluorophore that has an affinity for the amyloid fibrils. 20 Amyloid formation was quantitatively evaluated by the fluorescence emission of Thioflavine T (ThT) bound to amyloid fibrils, as reported by Levine 2 6 and also Soto et al 27 Aliquots of Af31l-42 (a synthetic peptide with the same sequence as the one deposited in amyloid plaques in Alzheimer's brain) at a concentration of 0.5 mg/ml prepared in 0.1M Tris, pH 7.4 were WO 2004/005336 PCT/EP2003/050287 20 incubated for 5 days at 37 0 C, gently swirled on a rotary shaker, in the absence or in the presence of different concentrations of the compounds. At the end of the incubation period, 50 mM glycine, pH 9.2 and 2 ItM ThT were added in a final volume of 2 ml. Fluorescence was measured at excitation 435 nm and emission 485 nm in a Perkin Elmer, model LS50B 5 fluorescence spectrometer. Using the analytical method 28 , the percentage of inhibition of fibril formation caused by compounds of the invention can be calculated. The results are listed in Table 4 and shown graphically in Figure 1. Values correspond to the percentage of inhibition of amyloid formation and are expressed relative to the activity of the compound of Example 8 (100%). 10 Table 4. Inhibition of AI 1
-
42 fibril formation by compounds of the invention Example n' % Inhibition (Compound of Example 8 as 100%) 1 88 4 80 5 80 6 85 Cellular assay of activity. Amyloid fibrils are cytotoxic, inducing cell death by apoptosis.' 8 The ability of the 15 compounds of the invention in preventing the amyloid formation can be evaluated by measuring the inhibition of the cytotoxicity in a cell assay. Aliquots of AP3 14 2 at a concentration of 0.5 mg/ml prepared in 0.1M Tris, pH 7.4 were incubated alone or in the presence of different concentrations of the compounds for 36h at 37 0 C, gently swirled on a rotary shaker. At the end of the incubation period, an aliquot of the 20 solution was added to the medium of PC 12 cells to reach a final concentration of AP3 of 5.5 p M. The cells were incubated for 24h and thereafter the cellular viability was evaluated using the MTT kit (Roche, Mannheim, Germany) according to the manufacturer's instructions. The results, listed in Table 5, are reported as the percentage of inhibition of WO 2004/005336 PCT/EP2003/050287 21 amyloid cytotoxicity, versus A3 incubated alone and expressed relative to the activity of the compound of Example 8 (100%). Table 5. Inhibition of amyloid cytotoxicity for compounds of the invention Example no % inhibition of cell death (Compound of Example 8 as 100%) 1 150 6 80 5 Blood-brain barrier permeability studies. The ability of compound of the invention to cross the BBB can be checked by capillary depletion experiments and the kinetics of their penetration into the brain can be measured. 10 a) Capillary depletion and blood washout Capillary depletion is used to assess the penetration into the brain of compounds of the invention. A "wash-out" step removes blood from the brain so that levels of the compounds of the invention present in the brain parenchyma can be measured. 15 Capillary depletion studies 27 were done in male CD-1 mice (28-36g). Mice are anaesthetized with i.p. urethane (40%) and the left jugular vein is exposed. A tritium labelled peptide of example 15 is injected i.v. . Before sacrificing the animals, blood is collected from the carotid artery (group 1) or from the descending aorta (group 2). After collection of blood, mice of group 1 are sacrificed or blood is removed by injecting 20 ml lactated Ringer's 20 solution (7.19 g/l NaC1, 0.3 g/1 KCL, 0.28 CaCl2, 2.1 g/l NaHCO 3 , 0.16+ g/l KH 2
PO
4 , 0.37 g/1 MgCl 2 *6H20, 0.99 g/1 D-glucose, 10 g/1 bovine serum albumin, pH 7.4) into the left ventricle of the heart for 30 sec, which removes more than 95% of the vascular contents of the brain (blood brain washout, group 2). The brain/serum ratio (pl/g) is evaluated by the equation: Brain/serum ratio = (cpm/g 25 brain)/(cpm/pl serum). The cerebral cortex is weighed and homogenized in a physiological WO 2004/005336 PCT/EP2003/050287 22 buffer (10 mM HEPES, 140 mM NaCl, 4 mM KCI, 2.8 mM CaC1 2 , 1 mM MgSO 4 , 1 mM Na
H
2
PO
4 , and 10 mM D-glucose, pH 7.4). Dextran solution (1.6 ml of a 26% solution) is then added to the homogenate. After centrifugation (5,400 g, 15 min, 4 0 C), brain vasculature (pellet) and parenchyma (supernatant) are separated and the radioactivity determined in each 5 fraction. After 10 min post i.v injection of radioactive derivative of Example 1 (compound of Example 7), more than 70% of the radioactivity is recovered in the parenchyma. The percentage of radioactivity associated with the capillary fraction with or without blood wash 10 out is 24 % and 29%, respectively. These data show that the majority of compound 1 crosses the BBB. In addition, it proves that the peptide of Example 1 is weakly binding to the luminal surface of the capillaries and is not sequestrated into the endothelial cells. b) Blood brain barrier permeability study: 15 The kinetics of penetration of compound of the invention into the brain can be evaluated through blood brain barrier permeability experiments. The percentage of injected peptide found in the brain can then be calculated. Mice are anaesthetized with i.p. urethane (40%) and the left jugular vein is exposed. 0.2 ml 20 of lactate Ringer's solution (7.19 g/1 NaC1, 0.3 g/1 KCL, 0.28 CaCl2, 2.1 g/1 NaHCO 3 , 0.16+ g/l KH 2
PO
4 , 0.37 g/1 MgCl 2 6H 2 0, 0.99 g/1 D-glucose, 10 g/l bovine serum albumin, pH 7.4) containing 1% BSA and tritium labelled peptide ("hot") of Example 7 (100 000 cpm/ml of tritium radioactive peptide) is injected. Arterial blood is collected from the right carotid artery at different time points following the labelled peptide injection. Serum is obtained by 25 centrifugation (4800 g, 10 min, 4oC). Following arterial blood collection, the mice are decapitated and the whole brains, except the pineals and pituitaries, are harvested and weighed. The amounts of radioactivity in brain and serum are determined after an overnight solubilization step in TS-2 solution (RPI, Mount Prospect, IL) at 40 0 C. The brain serum ratio of total radioactivity was determined over time from 1 min up to 120 min after injection. The 30 brain to serum ratio (gl/g) is estimated by the equation: Brain/serum ratio= (cpm/g brain)/(cpm/p1l serum).
WO 2004/005336 PCT/EP2003/050287 23 The representation of the brain to serum ratio versus time allows the derivation of the influx rate, Ki (slope) and the volume of distribution (Y intercept), Vi. The influx rate (Ki, microl (serum)/g (tissue weight)-min) represents the rate at which compounds move firomn the circulation to the brain. The volume of distribution (Vi, microl (seruim)/g (tissue weight)) is 5 the apparent volume of material which is distributed to the brain parenchyma at time 0 min. Results are presented in Table 6. Table 6. Brain uptake study of compound of Example 7 Injection (i.v.) Ki VI composition (microl/g-min) (microl/g) Hot* 3.54 +/- 0.29 50.6 +/- 12.4 *Hot: Radioactively labeled compound 10 Ki: influx rate Vi: volume of distribution The influx rate (Ki), with radioactive peptide, alone was 3.54 +/- 0.29 microl/g-min with a Y intercept of about 50 microl/g (Table 4). This indicates that compound of Example 1 15 penetrates the brain. The percentage of injected dose of compound of example 7, taken up per g of brain, was calculated (see Figure 2). The maximum value was 0.464% and half of this value was reached 1.4 min after injection. This data suggests a rapid brain uptake of compound of Example 7. 20 HPLC analysis of brain samples showed that 12% of intact peptide was recovered 20 min post i.v. injection. More than 0.06% of the injected compound was recovered intact in the brain within the 20 min post injection. 25 WO 2004/005336 PCT/EP2003/050287 24 In vivo studies using an animal model of cerebral AD deposition. The inhibitory activity the compound of the invention in the formation of amyloid deposits 5 by can be visualized in vivo by staining cerebral tissue sections with anti-AP31.42 antibodies in the presence and absence of a peptide of the invention. Male Fischer-344 rats weighed 250-300g and were 3-4 months of age at the time of arrival. The animals were housed 2 per cage, maintained on a 12 h light-dark cycle with access to 10 food and water ad libitum and were habituated to their new environment for 2-3 weeks prior to surgery. Surgery was performed under sodium pentobarbital (50 mg/kg, i.p.) anaesthesia. Atropine sulphate (0.4 mg/kg) and ampicillin sodium salt (50 mg/kg) were injected sub cutaneously once the animals were anaesthetized. API3-42 was dissolved in dimethylsulfoxide (DMSO) and then diluted with water to a concentration of 16.7% DMSO. The animal 15 received a bilateral injection of 5.0 nmol AP 14 2 into each amygdale by using a Kopf stereotaxic instrument with the incisor bar set at 3.3 mm below the interaural line. Injection coordinates measured from the bregma and the surface of the skull (AP -3.0, ML ± 4.6, DV 8.8) were empirically determined based on the atlas of Paxinos and Watson. A volume of 3.0 tl of the solution of AP3 1
-
4 2 at 5.0 nmol was administered over 6 min (flow rate 0.5 pl/min) 20 using a CMA/100 micro syringe pump. The cannula was left in situ for 2 min following injection, then it was withdrawn 0.2 mm and left for 3 min, and following these 5 min the camnnula was slowly withdrawn. Following surgery the animals were placed on a heating pad until they regained their righting reflex. The animals were treated with compound of Example 1 and of Example 8. The peptides, solubilized in a 0.9% NaCl at the concentration 25 of 4.4 mM were injected s.c (0.5 ml per injection), 4 times a week during 7 and a half weeks. After treatment with the compounds, animals were sacrificed by an overdose of sodium pentobarbital (150 mg/kg, i.p.), perfused transaortically. For histology, serial coronal sections (40 ptm) of the brain were cut, placed in ethylene glycol cryoprotectant and stored at -20°C until stained. Tissue sections were stained with anti-AP3l-42 antibodies, as described. 30 An image analysis system was used to determine the size of the amyloid deposits. These data were analysed by a two-way ANOVA followed by a Newman-Keuls' multiple range test for post hoc comparisons.
WO 2004/005336 PCT/EP2003/050287 25 The photographic results for the compound of Example 1 in comparison with vehicle and the compound of comparative Example 8 are shown in Figure 3A. Graphical results depicting the amyloid area are shown in Figure 3B. It is clear that the compound of Example 1 substantially reduces the amyloid area. 1 Selkoe, Science 275 1997, 630-631; 2 Masters, C. et al. Proc. Natl. Acad. Sci. USA 82 1985, 4245-4249; 3 Davies, L. et al. Neurology 38 1988, 1688-1693; 4 Kowall, N. W. et al. Proc. Natl. Acad. Sci. USA 88 1991, 7247-7251; 5 Levy, E. et al. Science 248 1990, 1124-1126; 6 Maury, C.P. Lab Invest. 72 1995, 4-16; 7 Jordan, B.D. Semin. Neurol. 20 2001,79-85; 8 Kang, J. et al. Nature 325 1987, 733-; 9 Mann, D.M; Histopathology 13 1988,125-37; 10 Shoji, M. et al. Science 258 1992, 126-; 11 Mori, H. et al. J. Biol. Chem. 267 1992, 17082-; 12 Soto J. Mol. Med. 77 1999, 412-418; 13 Serpell et al. Cell Mol. Life Sci. 53 1997, 871-887; 14 Soto, C. et al. J. Biol. Chem. 270 1995, 3063-3067; 15 Wisniewski et al. Biochemical Society Transactions 30 (4) 2002, 574-578; 16 WO 96/39834 (New York University); 17 WO 01/34631 (Axonyx Inc.); 18 WO 01/07473 (Stott, K.); 19 WO 02/074931 University of Chicago; 20 Yankner, B.A.; Neuron 16 1996, 921-932; 21 Gennaro, A.R., et al., Remington's Pharmaceutical Sciences, Part 8; 18th ed. Easton: The Mack Publishing Company, 1995; 22 Oral Sustained Release Formulations: Design and Evaluation; by Avraham Yacobi, Eva Halperin-Walega (Editor); 1st Ed. edition; Pergamon Press (April 1988); 23 Sustained-Release Injectable Products; Judy Senior (Editor); Interphann Press (July 2000); 24 Encapsulation and Controlled Release by D.R. Karsa, R.A. Stephenson (Editor) Springer Verlag (December 1993); WO 2004/005336 PCT/EP2003/050287 26 25 Principles of Peptide Synthesis (Springer Laboratory); Miklos Bodanszky, Milkos; Bodanszky; 2nd revised edition; Springer Verlag; 26 Levine, H; Protein science, 2 (3) 1993, 404-410; 27 Triguero, D. et al. J. Neurochem. 6 1990, 1882-1888;
Claims (19)
1. A compound of the general Formula I: 10 0 0 OH HOH NR 2 NR NR NR R H HH H - H o0 0 (I) 5 wherein: R' is selected from H and C 2 -C 6 -acyl; R 2 , R 3 , R 4 and R are independently selected from H and C 1 -C 6 -alkyl and at least one among R 2 , R 3 , R 4 and R 5 is C 1 -C 6 -alkyl; R 6 is selected from OH and NR 7 R 8 wherein R 7 and R 8 are independently selected from H 10 and C 1 -C 6 -alkyl; and salts and tritiated derivatives thereof
2. A compound according to claim 1, wherein R' is C 2 -C 6 -acyl.
3. A compound according to claim 1, wherein R 1 is selected from formyl, 15 acetyl, propionoyl and butyroyl.
4. A compound according to claim 1, wherein R' is acetyl.
5. A compound according to claim 1, wherein the stereochemistry at all the 20 a-carbon atoms is L;
6. A compound according to claims 1 to 5, wherein R 6 is NR 7 R 8 wherein R 7 and R are independently selected from H and C 1 -C 6 -alkyl and the stereochemistry at all the o-carbon atoms is L. WO 2004/005336 PCT/EP2003/050287 28
7. A compound according to any one of the preceding claims, wherein R is NH 2 . 5
8. A compound according to any one of the preceding claims wherein R 2 is H; R 4 and R 5 are independently selected from H and methyl.
9. A compound according to any one of the preceding claims, wherein R 3 is methyl. 10
10. A compound according to claim 9, wherein R 2 , R 4 and R s are H.
11. A compound of claim 9 wherein R 4 and R 5 are methyl; R 2 is H. 15
12. A compound of claim 9 wherein R 5 is methyl; R 2 and R 4 are H.
13. A compound of claim 9 wherein R 4 is methyl; R 2 and R 5 is H.
14. A compound according to any one of the preceding claims selected from the 20 following group: Ac-Leu-Pro-N(Me)-Phe-Phe-Asp-NH2 Ac-Leu-Pro-Phe-N(Me)-Phe-Asp-NH 2 25 Ac-Leu-Pro-Phe-N(Me)-Phe-N(Me)-Asp-NH2 Ac-Leu-Pro-N(Me)-Phe-Phe-N(Me)-Asp-NH 2 30 Ac-Leu-Pro-N(Me)-Phe-N(Me)-Phe-Asp-NH2 Ac-Leu-Pro-N(Me)-Phe-N(Me)-Phe-N(Me)-Asp-NH2 WO 2004/005336 PCT/EP2003/050287 29 Ac-Leu- [3 H]Pro-N(Me)-Phe-Phe-Asp-NH2
15. A compound according to any one of the preceding claims for use as a medicament. 5
16. A pharmaceutical composition comprising a compound according to any one of claims 1 to 14, and a pharmaceutically acceptable excipient, diluent or carrier.
17. Use of a compound according to any one of claims I to 14 for the 10 manufacture of a medicament for the treatment or prevention of a disease or condition selected from Alzheimer's disease, Dementia pugilistica (including head trauma), Hereditary Cerebral Haemorrhage with amyloidosis of the Dutch type (HCHWA-D) and vascular dementia with amyloid angiopathy. 15
18. Use according to claim 17, wherein the disease is Alzheimer's disease.
19. Use of a compound according to any one of claims 1 to 14, for the preparation of a medicament for the treatment of a disease associated with abnormal protein folding into amyloid and amrnyloid-like deposits.
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP02100787.7 | 2002-07-08 | ||
EP02100787 | 2002-07-08 | ||
EP02102834.5 | 2002-12-19 | ||
EP02102834 | 2002-12-19 | ||
PCT/EP2003/050287 WO2004005336A2 (en) | 2002-07-08 | 2003-07-07 | β-SHEET BREAKING PEPTIDES |
Publications (1)
Publication Number | Publication Date |
---|---|
AU2003271745A1 true AU2003271745A1 (en) | 2004-01-23 |
Family
ID=30116915
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU2003271745A Abandoned AU2003271745A1 (en) | 2002-07-08 | 2003-07-07 | Beta-SHEET BREAKING PEPTIDES |
Country Status (7)
Country | Link |
---|---|
EP (1) | EP1532170A2 (en) |
JP (1) | JP2006508904A (en) |
AU (1) | AU2003271745A1 (en) |
CA (1) | CA2489754A1 (en) |
IL (1) | IL166102A0 (en) |
NO (1) | NO20050436D0 (en) |
WO (1) | WO2004005336A2 (en) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20130065042A1 (en) | 2011-03-11 | 2013-03-14 | The Board Of Trustees Of The University Of Illinois | Micro-Vascular Materials And Composites For Forming The Materials |
SG187271A1 (en) * | 2011-07-07 | 2013-02-28 | Agency Science Tech & Res | Anti-amyloidogenic, alpha-helix breaking ultra-small peptide therapeutic |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB9917724D0 (en) * | 1999-07-28 | 1999-09-29 | Medical Res Council | Peptides |
IL149392A0 (en) * | 1999-11-05 | 2002-11-10 | Axonyx Inc | PEPTIDE ANALOGS AND MIMETICS SUITABLE FOR IN VIVO USE IN THE TREATMENT OF DISEASES ASSOCIATED WITH ABNORMAL PROTEIN FOLDING INTO AMYLOID, AMYLOID-LIKE DEPOSITS OR β-SHEET RICH PATHOLOGICAL PRECURSOR THEREOF |
-
2003
- 2003-07-07 WO PCT/EP2003/050287 patent/WO2004005336A2/en active Application Filing
- 2003-07-07 JP JP2004518784A patent/JP2006508904A/en active Pending
- 2003-07-07 AU AU2003271745A patent/AU2003271745A1/en not_active Abandoned
- 2003-07-07 CA CA002489754A patent/CA2489754A1/en not_active Abandoned
- 2003-07-07 EP EP03753576A patent/EP1532170A2/en not_active Withdrawn
-
2005
- 2005-01-03 IL IL16610205A patent/IL166102A0/en unknown
- 2005-01-26 NO NO20050436A patent/NO20050436D0/en not_active Application Discontinuation
Also Published As
Publication number | Publication date |
---|---|
NO20050436L (en) | 2005-01-26 |
JP2006508904A (en) | 2006-03-16 |
WO2004005336A3 (en) | 2004-02-26 |
CA2489754A1 (en) | 2004-01-15 |
NO20050436D0 (en) | 2005-01-26 |
WO2004005336A2 (en) | 2004-01-15 |
EP1532170A2 (en) | 2005-05-25 |
IL166102A0 (en) | 2006-01-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP0843516B1 (en) | Peptides and pharmaceutical compositions thereof for treatment of disorders or diseases associated with abnormal protein folding into amyloid or amyloid-like deposits | |
Kapurniotu et al. | Structure-based design and study of non-amyloidogenic, double N-methylated IAPP amyloid core sequences as inhibitors of IAPP amyloid formation and cytotoxicity | |
US20060069058A1 (en) | Beta-sheet breaker peptide analogs that inhibit beta-pleated sheet formation in amyloid beta-peptide | |
US5985242A (en) | Modulators of β-amyloid peptide aggregation comprising D-amino acids | |
RU2260599C2 (en) | MODULATING AGENT FOR AGGREGATION OF β-AMYLOID FOR AGGREGATION INHIBITION OF NATURAL β-AMYLOID OR FOR TREATMENT OF SUBJECT WITH DISORDER ASSOCIATED WITH β-AMYLOIDOSIS, PHARMACEUTICAL COMPOSITION AND METHOD FOR DETECTION OF NATURAL β-AMYLOID PEPTIDE IN BIOLOGICAL SAMPLE | |
AU781044B2 (en) | Peptide analogs and mimetics suitable for in vivo use in the treatment of diseases associated with abnormal protein folding into amyloid, amyloid-like deposits or beta-sheet rich pathological precursor thereof | |
AU4238797A (en) | Modulators of beta-amyloid peptide aggregation comprising d-amino acids | |
HU221629B1 (en) | Peptides as inhibitors of beta-amyloid protein production, process for their preparation and pharmaceutical compositions comprising thereof | |
JPH04506803A (en) | Small cyclic platelet aggregation inhibitor | |
JP2003503312A (en) | Stereoselective antigen fibril forming peptide and its peptidomimetic | |
AU2006215406A1 (en) | Amyloid-binding peptides, analogues and uses thereof | |
JPH07304795A (en) | Cyclopeptide of general formula i | |
KR20000016700A (en) | Depsipeptides and drugs containing the same as the active ingredient | |
US7060671B1 (en) | Peptides containing N-substituted D-amino acids for preventing β-strand association | |
AU2003271745A1 (en) | Beta-SHEET BREAKING PEPTIDES | |
US20060281686A1 (en) | Aza-peptides | |
US20070293422A1 (en) | Beta-Amyloid Inhibitors and Use Thereof | |
MXPA96005870A (en) | Endothelin antagonists ii |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PC1 | Assignment before grant (sect. 113) |
Owner name: LABORATOIRES SERONO SA Free format text: FORMER APPLICANT(S): APPLIED RESEARCH SYSTEMS ARS HOLDING N.V. |
|
MK4 | Application lapsed section 142(2)(d) - no continuation fee paid for the application |