CA2453050A1 - Degradable polyacetal polymers - Google Patents
Degradable polyacetal polymers Download PDFInfo
- Publication number
- CA2453050A1 CA2453050A1 CA002453050A CA2453050A CA2453050A1 CA 2453050 A1 CA2453050 A1 CA 2453050A1 CA 002453050 A CA002453050 A CA 002453050A CA 2453050 A CA2453050 A CA 2453050A CA 2453050 A1 CA2453050 A1 CA 2453050A1
- Authority
- CA
- Canada
- Prior art keywords
- group
- groups
- polymer
- aryl
- alkanediyl
- 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
- 229920000642 polymer Polymers 0.000 title claims abstract description 124
- 229920006324 polyoxymethylene Polymers 0.000 title abstract description 147
- 229930182556 Polyacetal Natural products 0.000 title abstract description 128
- 239000003814 drug Substances 0.000 claims abstract description 40
- 229940079593 drug Drugs 0.000 claims abstract description 31
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 12
- 229940041181 antineoplastic drug Drugs 0.000 claims abstract description 7
- 239000012867 bioactive agent Substances 0.000 claims description 62
- -1 cycloalkynyloxy Chemical group 0.000 claims description 49
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 35
- QYKIQEUNHZKYBP-UHFFFAOYSA-N Vinyl ether Chemical compound C=COC=C QYKIQEUNHZKYBP-UHFFFAOYSA-N 0.000 claims description 35
- 125000003118 aryl group Chemical group 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 28
- 229910052739 hydrogen Inorganic materials 0.000 claims description 27
- 125000002877 alkyl aryl group Chemical group 0.000 claims description 26
- 239000001257 hydrogen Substances 0.000 claims description 26
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims description 26
- 125000003710 aryl alkyl group Chemical group 0.000 claims description 25
- 125000000217 alkyl group Chemical group 0.000 claims description 24
- 238000002360 preparation method Methods 0.000 claims description 23
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 claims description 20
- 125000003342 alkenyl group Chemical group 0.000 claims description 20
- 125000000304 alkynyl group Chemical group 0.000 claims description 19
- 238000006116 polymerization reaction Methods 0.000 claims description 18
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 18
- 230000008569 process Effects 0.000 claims description 16
- 239000000580 polymer-drug conjugate Substances 0.000 claims description 15
- 230000003213 activating effect Effects 0.000 claims description 14
- 150000002009 diols Chemical class 0.000 claims description 14
- 239000008194 pharmaceutical composition Substances 0.000 claims description 14
- 125000006239 protecting group Chemical group 0.000 claims description 14
- 125000004432 carbon atom Chemical group C* 0.000 claims description 13
- 229960000834 vinyl ether Drugs 0.000 claims description 12
- 229960004679 doxorubicin Drugs 0.000 claims description 11
- 150000004676 glycans Chemical class 0.000 claims description 11
- 125000004430 oxygen atom Chemical group O* 0.000 claims description 11
- 229920001282 polysaccharide Polymers 0.000 claims description 11
- 239000005017 polysaccharide Substances 0.000 claims description 11
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 10
- 150000001720 carbohydrates Chemical class 0.000 claims description 8
- 239000003937 drug carrier Substances 0.000 claims description 8
- 125000004442 acylamino group Chemical group 0.000 claims description 7
- 125000004423 acyloxy group Chemical group 0.000 claims description 7
- 125000005041 acyloxyalkyl group Chemical group 0.000 claims description 7
- 125000003302 alkenyloxy group Chemical group 0.000 claims description 7
- 125000004183 alkoxy alkyl group Chemical group 0.000 claims description 7
- 125000004171 alkoxy aryl group Chemical group 0.000 claims description 7
- 125000003545 alkoxy group Chemical group 0.000 claims description 7
- 125000005083 alkoxyalkoxy group Chemical group 0.000 claims description 7
- 125000003282 alkyl amino group Chemical group 0.000 claims description 7
- 125000005133 alkynyloxy group Chemical group 0.000 claims description 7
- 125000000266 alpha-aminoacyl group Chemical group 0.000 claims description 7
- 125000002431 aminoalkoxy group Chemical group 0.000 claims description 7
- 125000004103 aminoalkyl group Chemical group 0.000 claims description 7
- 125000005128 aryl amino alkyl group Chemical group 0.000 claims description 7
- 125000000000 cycloalkoxy group Chemical group 0.000 claims description 7
- 238000006073 displacement reaction Methods 0.000 claims description 7
- 125000004438 haloalkoxy group Chemical group 0.000 claims description 7
- 125000001188 haloalkyl group Chemical group 0.000 claims description 7
- 125000003106 haloaryl group Chemical group 0.000 claims description 7
- 150000004702 methyl esters Chemical class 0.000 claims description 7
- 125000004945 acylaminoalkyl group Chemical group 0.000 claims description 6
- 125000004465 cycloalkenyloxy group Chemical group 0.000 claims description 6
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 claims description 5
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 claims description 5
- WEAHRLBPCANXCN-UHFFFAOYSA-N Daunomycin Natural products CCC1(O)CC(OC2CC(N)C(O)C(C)O2)c3cc4C(=O)c5c(OC)cccc5C(=O)c4c(O)c3C1 WEAHRLBPCANXCN-UHFFFAOYSA-N 0.000 claims description 4
- 150000001298 alcohols Chemical class 0.000 claims description 4
- 239000003054 catalyst Substances 0.000 claims description 4
- 229920001542 oligosaccharide Polymers 0.000 claims description 4
- 150000002482 oligosaccharides Chemical class 0.000 claims description 4
- 230000001093 anti-cancer Effects 0.000 claims description 3
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 claims description 2
- 229930012538 Paclitaxel Natural products 0.000 claims description 2
- 150000001338 aliphatic hydrocarbons Chemical class 0.000 claims description 2
- 150000004945 aromatic hydrocarbons Chemical class 0.000 claims description 2
- 150000004292 cyclic ethers Chemical class 0.000 claims description 2
- 150000002170 ethers Chemical class 0.000 claims description 2
- 229940052303 ethers for general anesthesia Drugs 0.000 claims description 2
- 239000003960 organic solvent Substances 0.000 claims description 2
- 229960001592 paclitaxel Drugs 0.000 claims description 2
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 claims description 2
- 229940063683 taxotere Drugs 0.000 claims description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 2
- 125000004178 (C1-C4) alkyl group Chemical group 0.000 claims 1
- 125000006656 (C2-C4) alkenyl group Chemical group 0.000 claims 1
- 125000000467 secondary amino group Chemical class [H]N([*:1])[*:2] 0.000 claims 1
- 125000001302 tertiary amino group Chemical group 0.000 claims 1
- 206010028980 Neoplasm Diseases 0.000 abstract description 12
- 201000011510 cancer Diseases 0.000 abstract description 10
- 230000002349 favourable effect Effects 0.000 abstract description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 51
- 239000000243 solution Substances 0.000 description 40
- 239000000562 conjugate Substances 0.000 description 39
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 36
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 35
- 239000000203 mixture Substances 0.000 description 30
- 229920001223 polyethylene glycol Polymers 0.000 description 29
- 210000003743 erythrocyte Anatomy 0.000 description 24
- 230000021615 conjugation Effects 0.000 description 20
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- 230000015572 biosynthetic process Effects 0.000 description 16
- 230000009089 cytolysis Effects 0.000 description 15
- 238000003786 synthesis reaction Methods 0.000 description 15
- 239000002202 Polyethylene glycol Substances 0.000 description 14
- 230000000975 bioactive effect Effects 0.000 description 13
- 230000015556 catabolic process Effects 0.000 description 13
- 238000006243 chemical reaction Methods 0.000 description 13
- 238000006731 degradation reaction Methods 0.000 description 13
- 235000018102 proteins Nutrition 0.000 description 13
- 102000004169 proteins and genes Human genes 0.000 description 13
- 108090000623 proteins and genes Proteins 0.000 description 13
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 12
- 238000003556 assay Methods 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 12
- 229920006237 degradable polymer Polymers 0.000 description 12
- 229920001577 copolymer Polymers 0.000 description 11
- 201000010099 disease Diseases 0.000 description 11
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 11
- 238000005227 gel permeation chromatography Methods 0.000 description 11
- 239000000178 monomer Substances 0.000 description 11
- 239000011541 reaction mixture Substances 0.000 description 11
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 11
- 150000001875 compounds Chemical class 0.000 description 10
- 239000004215 Carbon black (E152) Substances 0.000 description 9
- 229920002307 Dextran Polymers 0.000 description 9
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 239000003795 chemical substances by application Substances 0.000 description 9
- 231100000135 cytotoxicity Toxicity 0.000 description 9
- 229930195733 hydrocarbon Natural products 0.000 description 9
- 239000000546 pharmaceutical excipient Substances 0.000 description 9
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 9
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 8
- 210000004369 blood Anatomy 0.000 description 8
- 239000008280 blood Substances 0.000 description 8
- 239000011203 carbon fibre reinforced carbon Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 238000001727 in vivo Methods 0.000 description 8
- 229920002873 Polyethylenimine Polymers 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 7
- 239000000969 carrier Substances 0.000 description 7
- 230000003013 cytotoxicity Effects 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 108090000765 processed proteins & peptides Proteins 0.000 description 7
- 239000007787 solid Substances 0.000 description 7
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 6
- 230000004087 circulation Effects 0.000 description 6
- 125000000392 cycloalkenyl group Chemical group 0.000 description 6
- 231100000433 cytotoxic Toxicity 0.000 description 6
- 230000001472 cytotoxic effect Effects 0.000 description 6
- 238000009826 distribution Methods 0.000 description 6
- 150000002148 esters Chemical class 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 239000000725 suspension Substances 0.000 description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- 150000001241 acetals Chemical class 0.000 description 5
- 230000002378 acidificating effect Effects 0.000 description 5
- 229940024606 amino acid Drugs 0.000 description 5
- 235000001014 amino acid Nutrition 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 5
- 230000006037 cell lysis Effects 0.000 description 5
- 125000000753 cycloalkyl group Chemical group 0.000 description 5
- 239000007857 degradation product Substances 0.000 description 5
- 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 5
- 238000000338 in vitro Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 210000004185 liver Anatomy 0.000 description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 210000000056 organ Anatomy 0.000 description 5
- 239000002953 phosphate buffered saline Substances 0.000 description 5
- 239000002243 precursor Substances 0.000 description 5
- 102000004196 processed proteins & peptides Human genes 0.000 description 5
- 229920006395 saturated elastomer Polymers 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- KYRUKRFVOACELK-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 3-(4-hydroxyphenyl)propanoate Chemical compound C1=CC(O)=CC=C1CCC(=O)ON1C(=O)CCC1=O KYRUKRFVOACELK-UHFFFAOYSA-N 0.000 description 4
- YSFGBPCBPNVLOK-UHFFFAOYSA-N 6-hydroxy-2-methylhex-2-enamide Chemical compound NC(=O)C(C)=CCCCO YSFGBPCBPNVLOK-UHFFFAOYSA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 4
- 108010039918 Polylysine Proteins 0.000 description 4
- 229920000615 alginic acid Polymers 0.000 description 4
- 235000010443 alginic acid Nutrition 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000000259 anti-tumor effect Effects 0.000 description 4
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- 229920000656 polylysine Polymers 0.000 description 4
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 4
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- CYIGRWUIQAVBFG-UHFFFAOYSA-N 1,2-bis(2-ethenoxyethoxy)ethane Chemical compound C=COCCOCCOCCOC=C CYIGRWUIQAVBFG-UHFFFAOYSA-N 0.000 description 3
- JPVNTYZOJCDQBK-UHFFFAOYSA-N 3-ethenoxypropan-1-amine Chemical compound NCCCOC=C JPVNTYZOJCDQBK-UHFFFAOYSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
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- 239000001990 protein-drug conjugate Substances 0.000 description 1
- 235000019423 pullulan Nutrition 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 125000005493 quinolyl group Chemical group 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 150000003333 secondary alcohols Chemical class 0.000 description 1
- 150000003335 secondary amines Chemical class 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 238000001542 size-exclusion chromatography Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000003381 stabilizer Substances 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
- 239000005720 sucrose Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 229920002994 synthetic fiber Polymers 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 150000003509 tertiary alcohols Chemical class 0.000 description 1
- 150000003512 tertiary amines Chemical group 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 125000001544 thienyl group Chemical group 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 239000011732 tocopherol Substances 0.000 description 1
- 235000010384 tocopherol Nutrition 0.000 description 1
- 229960001295 tocopherol Drugs 0.000 description 1
- 229930003799 tocopherol Natural products 0.000 description 1
- 125000003944 tolyl group Chemical group 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000003420 transacetalization reaction Methods 0.000 description 1
- ILWRPSCZWQJDMK-UHFFFAOYSA-N triethylazanium;chloride Chemical compound Cl.CCN(CC)CC ILWRPSCZWQJDMK-UHFFFAOYSA-N 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 125000001493 tyrosinyl group Chemical group [H]OC1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G2/00—Addition polymers of aldehydes or cyclic oligomers thereof or of ketones; Addition copolymers thereof with less than 50 molar percent of other substances
- C08G2/30—Chemical modification by after-treatment
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
- A61K47/645—Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G4/00—Condensation polymers of aldehydes or ketones with polyalcohols; Addition polymers of heterocyclic oxygen compounds containing in the ring at least once the grouping —O—C—O—
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L59/00—Compositions of polyacetals; Compositions of derivatives of polyacetals
Abstract
Degradable polyacetal polymers and functionalized degradable polyacetal polymers have properties favorable for use in pharmaceutical and biomedical applications. The degradable polyacetal polymers are relatively stable at physiological pH with favorable biodistribution profiles, and degrade readily in low pH conditions. Conjugates of the polymers with drugs, especially anticancer drugs, and use for treatment of cancer.
Description
DEGRADABLE POLYACETAL POLYMERS
BACKGROUND TO THE INVENTION
FIELD OF THE INVENTION
This invention relates to degradable polyacetal polymers and therapeutic agents derived therefrom, the production of these materials, and methods of disease treatment using them.
DESCRIPTION OF THE RELATED ART
Polymer therapeutics (R. Duncan, "Polymer therapeutics for tumor specific delivery", Chem. & Ihd., 7, 262-264, 1997) are developed for biomedical applications requiring physiologically soluble polymers; and include biologically active polymers, polymer-drug conjugates, polymer-protein conjugates, and other covalent constructs of bioactive molecules.
An exemplary class of a polymer-drug conjugate is derived from copolymers of hydroxypropyl methacrylamide (HPMA), which have been extensively studied for the conjugation of cytotoxic drugs for cancer chemotherapy (R. Duncan, "Drug-polymer conjugates: potential for improved chemotherapy", A~rti-Cancer Drugs, 3, 175-210, 1992; D.
Putnam et al., "Polymer conjugates with anticancer activity", Adv. Polym.
Sci., 122, 55-123, 1995; R. Duncan et al., "The role of polymer conjugates in the diagnosis and treatment of cancer", STP PhaYma, 6, 237-263, 1996). An HPMA copolymer conjugated to doxorubicin, known as PK-l, is currently in Phase II evaluation in the UK. PK-1 displayed reduced toxicity compared to free doxorubicin in the Phase I studies (P. Vasey et al., "Phase I clinical and pharmacokinetic study of PKI (HPMA copolymer doxorubicin): first member of a new class of chemotherapeutic agents: drug-polymer conjugates" Clih.
Cahcef° Res., 5, 83-94.
1999). The maximum tolerated dose of PK-1 was 320 mg/ma, which is 4-5 times higher than the usual clinical dose of free doxorubicin. .
The polymers used to develop polymer therapeutics may also be separately developed for other biomedical applications that require the polymer be used as a material. Thus, drug release matrices (including microparticles and nanoparticles), hydrogels (including injectable gels and viscous solutions) and hybrid systems (e.g. liposomes with conjugated polyethylene glycol) on the outer surface) and devices (including rods, pellets, capsules, films, gels) can be fabricated for tissue or site specific drug delivery. Polymers are also clinically widely used as excipients in drug formulation. Within these three broad application areas:
(1) physiologically S soluble molecules, (2) materials, and (3) excipients, biomedical polymers provide a broad technology platform for optimizing the efficacy of an active therapeutic drug.
An increasing number of physiologically soluble polymers have been used as macromolecular partners for the conjugation of bioactive molecules. Many polymers have the disadvantage of being non-degradable in the polymer backbone. For example, polyethylene glycol) (C. Monfardini et al., "Stabilization of substances in circulation", Bioconjugate Chem., 9, 418-450, 1998; S. Zalipsky, "Chemistry of polyethylene glycol conjugates with biologically active molecules", Adv. D~ugDelive~y Rev, 16,-1S7-182, 1995; C.
Delgado et al., "The uses and properties of PEG-liked proteins", C~it. Rev. They. Drug CaYrie~ Syst., B, 249-304, 1992; M.L. Nucci et al., "The therapeutic values of polyethylene glycol)-modified proteins", Adv. Drug DeliveYy Rev. 6, 133-151, 1991; A. Nathan et al., Copolymers of lysine and polyethylene glycol: A new family of functionalized drug carriers", Bioconjugate Chem.
4, S4-62, 1993) and HPMA (D. Putnam et al., "Polymer conjugates with anticancer activity", Adv. Pol~m. Sci., 122, SS-123, 1995; and R. Duncan et al., "The role of polymer conjugates in the diagnosis and treatment of cancer", STP Pha~ma, 6, 237-263, 1996) copolymers have been extensively studied for conjugation. PEG is also generally used in the pharmaceutical industry as a formulation excipient. These hydrophilic polymers are soluble in physiological media, but their main disadvantage is that the polymer mainchain does not degrade in vivo.
Thus it is not possible to prohibit accumulation of these polymers in the body. Only polymers with a molecular weight lower than the renal threshold can be used for systemic 2S administration. It is imperative that for the systemic use of non-degradable polymers such as HPMA and PEG only molecules of a molecular weight which are readily cleared be administered or else long-term deleterious accumulation in healthy tissue will invariably result (L. Seymour et al., "Effect of molecular weight (Mw) of N-(2-hydroxypropyl)methacrylamide copolymers on body distributions and rate of excretion after subcutaneous, intraperitoneal and intravenous administration to rats", J. Biomed. Mate.
Res. 21, 341-1358, 1987; P. Sclmeider et al., "A review of drug-induced lysosomal disorders of the liver in man and laboratory animals", Microscopy Res. Tech. 36, 253-275, 1997; C.
Hall et al., "Experimental hypertension elicited by injections of methyl cellulose", Experientia 17, 544-454, 1961; C. Hall et al., "Macromolecular hypertension:
hypertensive cardiovascular disease from subcutaneously administered polyvinyl alcohol", Experientia 18, 38-40, 1962).
Although some natural polymers such as polysaccharides have the advantage of being degradable in vivo, e.g. dextran, they typically lack a strict structural uniformity and have the propensity upon chemical modification (i.e. conjugation of a bioactive molecule) to become inununogenic or non-degradable (J. Vercauteren et al., "Effect of the chemical modification of dextran on the degradation by dextranases", J. Bio. Comp. Polymers 5, 4-15, 1990; W.
Shalaby et al., "Chemical modification of proteins and polysaccharides and its effect on enzyme-catalyzed degradation", in: S. Shalaby, ed. Biomedical Polymers.
Designed to-degrade systems. New York: Hanser Publishers, 1994). Other polysaccharides which have been investigated for biomedical conjugation applications include chitosan (Y.
Ohya et al., "a-1,4-Polygalactosamine immobilised 5-fluorouracils through hexamethylene spacer groups via urea bonds", J. Cont. Rel.,17, 259-266, 1991), alginate (A. Al-Shamkhani et al., "Synthesis, controlled release properties and antitumor activity of alginate cis-aconityl daunomycin conjugates", Int. J. Pharm., 122, 107-119, 1995; S. Morgan et al., "Alginates as drug carriers: covalent attachment of alginates to therapeutic agents containing primary amine groups", Int. J. Pharm., 122, 121-128, 1995), hyaluronic acid (B.
Schechter et al., "Soluble polymers as carriers of cisplatinum", J. Cont. Rel., 10, 75-87, 1989), 6-O-carboxymethyl chitan (Y. Ohya et al., "In vivo and in vitro antitumor activity of CM-Chitin immobilized doxorubicins by lysosomal digestible tetrapeptide spacer groups", J. Bioact.
Compat. Polymers,10, 223-234, 1995) and 6-O-carboxymethyl pullulan (H. Nogusa et al., "Synthesis of carboxymethylpullulan peptide doxorubicin conjugates and their properties", Claem. Pharm. Bull., 43, 1931-1936, 1995).
Other natural polymers such as proteins can also be used to conjugate a bioactive molecule. For example albumin has been investigated as a protein used to conjugate a bioactive molecule (P. Balboni et al., "Activity of albumin conjugates of 5-fluorodeoxyuridine and cytosine arabinoside on poxviruses as a lysosomotropic antiviral chemotherapy", Nature, 264, 181-183, 1976; A. Trouet et al., "A covalent linkage between daunorubicin and proteins that is stable in serum and reversible by lysosomal hydrolases as required for a lysosomotropic drug-carrier conjugate. In vitro and in vivo studies", Proc.
Natl. Acad. Sci. ZISA, 79, 626-629, 1982; F. Dosio et al., "Preparation, characterization and properties in vitro and in vivo of a paclitaxel-albumin conjugate", J. Cont.
Rel., 47(3), 293-304, 1997; T. Yasuzawa et al, "Structural determination of the conjugate of human serum albumin with a mitomycin C derivative, IOW-2149, by matrix assisted laser desorption/ionization mass spectrometry", Biocohjugate Chena., 8, 391-399, 1997; A.
blunder et al., "Antitumor activity of methotrexate-albumin conjugates in rats bearing a Walker-256 carcinoma", Int. J. Cancer, 76, 884-890, 1998). The major limitations for using a protein to conjugate a bioactive compound include the propensity for inducing immunogenicity and non-specific degradation of the protein in vivo, and denaturation and irreversible alteration of the protein during preparation of the conjugate.
Other proteins such as transferrin, which binds to the transferrin receptor and thus have the potential to undergo receptor-mediated uptake (T. Tanaka et al., "Intracellular disposition and cytotoxicity of transferrin-mitomycin C conjugate in HL60 cells as a receptor-mediated drug targeting system", Biol. Pharm. Bull., 21(2), 147-152, 1998) and various immuno-conjugates (D. Gaal et al., "Low toxicity and high antitumor activity of daunomycin by conjugation to an immunopotential amphoteric branched polypeptide", Eur. J. Cayacer, 34(1), 155-16, 1998; P.
Trail et al., "Site-directed delivery of anthracyclines for the treatment of cancer", Drug Dev.
Res. 34, 196-209, 1995; E. Eno-Amooquaye et al., "Altered biodistribution of an antibody-enzyme conjugate modified with polyethylene glycol", Br. J. Cancer, 73, 1323-1327, 1996;
P. Flanagan et al., "Evaluation of antibody-[N-(2-hydroxypropyl)methacrylamide] copolymer conjugates as targetable drug-carriers. 2. Body distribution of anti Thy-1,2 antibody, anti-transferrin receptor antibody B3/25 and transferrin conjugates in DBAZ mice and activity of conjugates containing daunomycin against L1210 leukemia in vivo", J. Cont.
Rel., l8, 25-38, 1992; C. Springer et al., "Ablation of human choriocarcinoma xenografts in nude mice by antibody-directed enzyme prodrug therapy (ADEPT) with three novel compounds", Eur. J.
CajZCer,11, 1362-1366, 1991.) also have been investigated. Monodisperse molecular weight distribution is often claimed to be a significant advantage for using proteins to conjugate drugs, but this can only be useful if a single species of the protein-drug conjugate can be reproducibly prepared on adequate scale which stable on storage. This is generally not economically or technologically possible to achieve in practice. Thus, there is a need for degradable synthetic polymers developed for biomedical application, and specifically for conjugation applications, which can address the limitations inherent in the use of natural polymers for these applications.
Synthetic polymers which have been prepared and studied that are potentially degradable include polymers derived from amino acids (e.g. poly(glutamic acid) , poly[SN-(2-hydroxyethyl)-L-glutamine), (3-poly(2-hydroxyethyl aspartamide), poly(L-glutamic acid) and polylysine). These polymers when prepared for conjugation applications that require physiological solubility do not degrade in vivo to any extent within a time period of 10-100 hours. Additionally polymers and copolymers including pseudo-poly(amino acids) (K. James et al., "Pseudo-poly(amino acids: Examples for synthetic materials derived from natural metabolites", in: K. Paxk, ed., Coht~olled Drug Deliver: Challenges ahd Strategies, Washington, DC: American Chemical Society, 389-403, 1997) and polyesters such as copolymers of polylactic and poly(glycolic acid), poly(a or b-malic acid) (K.
Abdellaoui et al., "Metabolite-derived artificial polymers designed for drug targeting, cell penetration and bioresorption", Eur. J. Pha~m. Sei., 6, 61-73, 1998; T. Ouchi et al., "Synthesis and antitumor activity of conjugates of poly (a-malic acid) and 5-fluorouracil bound via ester, amide or carbamoyl bonds", J. Coyat. Rel., 12, 143-153, 1990), and block copolymers such as PEG-lysine (A. Nathan et al., "Copolymers of lysine and polyethylene glycol: A new family of functionalized drug carriers", Biocohjugate Chern., 4, 54-62, 1993.), poly(lysine citramide) (K. Abdellaoui et al, "Metabolite-derived artificial polymers designed for drug targeting, cell penetration and bioresorption", Eu~. J. PhaYna. Sci., 6, 61-73, 1998) and amino acid-PEG
derived block copolymers (G. Kwon et al., Block copolymer micelles as long-circulating drug vehicles", Adv. Drug Del. Rev., 16, 295-309, 1995; and V. Alakhov et al., "Block copolymeric biotransport carriers as versatile vehicles for drug delivery", Exp. Opih. Invest.
DYUgs, 7(9), 1453-1473, 1998) have also been investigated for conjugation.
Acetals are well known to be hydrolytically labile under mildly acidic conditions.
Thus, biomedical polymers possessing acetal linkages in the polymer mainchain may undergo enhanced rates of hydrolysis in biological environments that are mildly acidic compared to biological environments that are at neutral or basic pH. For example, soluble polyacetals that can conjugate a bioactive molecule are expected to degrade at enhanced rates at the acetal functionality during cellular uptake because of the increase in acidity during endocytosis.
Polyacetals will also display enhanced rates of hydrolysis in acidic regions of the gastrointestinal tract. Additionally polyacetals would be expected to degrade at enhanced rates at sites of diseased tissue that are mildly acidic (e.g. solid tumors).
Preparing polyacetals can be accomplished by acetal- or transacetalization reactions which result in the formation of a low molecular weight by-product (e.g. water or an alcohol).
Complete removal of such a by-product is necessary for reproducible polymerization and to ensure the polyacetal does not degrade on storage. Usually harsh conditions are required to obtain high molecular weight polymer. If functionalized monomers relevant for biomedical applications are used, such conditions can often lead to unspecif ed chemical changes in the monomer. Polyacetals can be prepared without generation of a small molecule which requires removal by cationic ring-opening polymerization using bicyclic acetals (L.
Torres et al., "A
new polymerization system for bicyclic acetals: Toward the controlled/"living"
cationic ring-opening polymerization of 6,8-dioxabicyclo[3.2.1] octane", Mac~orraolecules, 32, 6958-6962, 1999). These reaction conditions lack versatility because they require bicyclic acetal monomers that are difficult to prepare with a wide range of chemical functionality useful for conjugation applications.
Polyacetals can also be prepared without generation of a small molecule byproduct that requires removal by the reaction of diols and di-vinyl ethers using an acid catalyst, as described by Heller (J. Heller et al., "Preparation of polyacetals by the reaction of divinyl ethers and polyols", J. Polyp. Sci.: Polyfn. Lett. Ed.,18, 293-297, 1980; J.
Heller et al., "Polyacetal hydrogels formed from divinyl ethers and polyols", US Patent No.
4,713,441, 1987). Such polyacetals have uniform structure in that they are strictly alternating polymers of the A-B type. Uniform structure in biomedical polymer development is critical for optimization of the biological profile and to ensure the polymer meet regulatory requirements. The polymerization of diols and di-vinyl ethers occurs without the elimination of a small molecule under mild conditions. This is more efficient than polymerizations where there is a molecule (e.g. water or methanol) which must be removed.
Polyacetals suitable for conjugation can be prepared by utilization of suitably functionalized diols, di-vinyl ethers, and/or hydroxy-vinyl ether monomers. Thus it becomes possible to prepare polyacetals for conjugation that possess conjugation functionality on either the A or B
monomeric unit. Such structural uniformity is advantageous for controlling conjugation of bioactive molecules along the polymer mainchain.
The production of biodegradable polyacetals derived from polysaccharides which chemically has been described in WO 96/32419. This approach does not give polymeric materials displaying structural uniformity and suffers from the aforementioned limitations where chemical modification (i.e. conjugation of a bioactive molecule) often leads to the polysaccharide to become immunogenic or non-degradable. It is not possible to prepare polymeric materials displaying an alternating A-B structure, rather the structure of these polysaccharide derived polyacetals are too diverse to chemically analyze to the degree necessary to fulfill regulatory requirements.
The disclosures of these and other documents referred to throughout this application are incorporated herein by reference in their entirety.
SUMMARY OF THE INVENTION
The first aspect of the present invention relates to a class of new degradable polymers represented by Formula (I):
R R' ,Y
C~ IC~'O n wherein R and Rl are independently selected from the group consisting of hydrogen, Cl_ls alkyl, Cz_is alkenyl, Cz_ls alkynyl, C6_l8 aryl, C~-18 alkaryl and C~_ls aralkyl groups;
X is a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1_zoo a~~ediyl, C2_zoo alkenediyl, Cz-zoo a~3'nediyl, C6_zoo cycloalkanediyl, C6_zoo cycloalkenediyl, C6_200 _g_ cycloalkynediyl, Cg_ZOO arylenediyl, C6_2oo alkarylenediyl, C6_2oo aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2-10,000.
Another aspect of the invention relates to a bioactive agent, preferably a drug, conjugated to a degradable polymer of Formula (I).
Another aspect of the invention relates to the preparation of the new degradable polymers and new polymer therapeutics.
A further aspect of the invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a polymer-drug conjugate of the present invention in combination with one or more pharmaceutically acceptable carriers.
A further aspect of the invention relates to the use of a polymer-drug conjugate of the present invention in the preparation of a medicament for the treatment of a disease such as cancer.
A further aspect of the invention relates to a method of treatment for diseases such as cancer, comprising administering to a patient in need of such treatment a therapeutically effective amount of a polymer therapeutic of the present invention.
Yet a further aspect of the present invention relates to prepolymers useful for the preparation of the degradable polymers.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 is a graph showing the superposition of gel permeation chromatography traces for polyacetal 3 dissolved in solution at pH 7 and at 37 °C over a period of 21 days.
Figure 2 is a graph showing the superposition of gel permeation chromatography traces for polyacetal 3 dissolved in solution at pH 5.5 and at 37 °C
over a period of 21 days.
Figure 3 is a graph showing polyacetal 3 degradation shown as percent molecular weight loss at pH values of 7.4, 6.5 and 5.5 versus time.
_g_ Figure 4 is a graph showing red blood cell lysis assay of polyacetal 3.
Figure 5 is a graph showing the cytotoxicity of polyacetal 3 using the B16F10 cell line.
Figure 6 is a graph showing superimposed gel permeation chromatography (GPC) traces of polyacetal 22 from a phosphate buffer solution at pH 7.4 and a solution where the pH was adjusted to 1-2 by the addition of HCl.
Figure 7 is a graph showing the degradation profile of polyacetal 22 at pH 7.4 and 5.5 displaying the loss in molecular weight (Mw).
Figure 8a is a graph showing red blood cell (RBC) lysis of polyacetal 22 and its degradation products over a 1 hour time period . No RBC lysis was observed in this assay for the polyacetal.
Figure 8b is a graph showing red blood cell lysis of polyacetal 22 and its degradation products over a 5 hour time period. No RBC lysis was observed in this assay for the polyacetal.
Figure 9 is a graph showing cytotoxicity of polyacetal 22 using the B 16F10 cell line.
Polyacetal 22 does not display cytotoxicity in this assay.
Figure 10 is a graph showing the labeling efficiency with lasI labeled Bolton-Hunter reagent of polyacetal 22 to give conjugate polyacetal 23. This Figure shows the crude product.
Figure 11 is a graph showing the labeling efficiency with lasl labeled Bolton-Hunter reagent of polyacetal 22 to give conjugate polyacetal 23. This Figure shows the purified conjugate product.
Figure 12 is a graph showing the body distribution of radiolabeled polyacetal conjugate at 5 min and 1 hour. This shows that the polyacetal conjugate remains in the blood without accumulating in the organs shown.
DETAILED DESCRIPTION OF THE INVENTION
I. Definitions Terms used herein are based upon their recognized meanings and should be clearly understood by those skilled in the art.
The term "alkyl" refers to a straight or branched saturated monovalent hydrocarbon radical having the number of carbon atoms as indicated.
The term "alkenyl" refers to a straight or branched unsaturated monovalent hydrocarbon radical having the number of carbon atoms as indicated and the distinguishing feature of a carbon-carbon double bond.
The term "alkynyl" refers to a straight or branched unsaturated monovalent hydrocarbon radical having the number of carbon atoms as indicated and the distinguishing feature of a carbon-carbon triple bond.
The term "cycloalkyl" refers to a cyclic saturated monovalent hydrocarbon radical having the number of carbon atoms as indicated.
The terms "cycloalkenyl" and "cycloalkynyl" refer to cyclic unsaturated monovalent hydrocarbon radicals. A "cycloalkenyl" is characterized by a carbon-carbon double bond and a "cycloalkynyl" is characterized by a carbon-carbon triple bond.
The term "aryl" refers to a monovalent unsaturated aromatic carbocyclic radical having one or two rings, such as phenyl, naphthyl, indanyl or biphenyl, or to a monovalent unsaturated aromatic heterocyclic radical such as quinolyl, dihydroisoxazolyl, furanyl, imidazolyl, pyridyl, phthalimido, thienyl and the like.
The term "alkaryl" refers to an aryl group substituted with one or more alkyl groups.
The term "aralkyl" refers to an alkyl group substituted with one or more aryl groups.
The term "alkanediyl" refers to a straight or branched saturated divalent hydrocarbon radical having the number of carbon atoms indicated.
The terms "alkenediyl" and "alkynediyl" refer to straight or branched,unsaturated divalent hydrocarbon radicals. An "alkenediyl" is characterized by a carbon-carbon double bond and an "alkynediyl" is characterized by a carbon-carbon triple bond.
The term "cycloalkanediyl" refers to a cyclic saturated divalent hydrocarbon radical having the number of carbon atoms indicated.
The terms "cycloalkenediyl" and "cycloalkynediyl" refer to cyclic unsaturated divalent hydrocarbon radicals. A "cycloallcenediyl" is characterized by a carbon-carbon double bond and a "cycloalkynediyl" is characterized by a carbon-carbon triple bond.
The term "arylenediyl" refers to a divalent unsaturated aromatic carbocyclic radical having one or two rings. The term "alkarylenediyl" refers to an arylenediyl substituted with one or more alkyl groups and the term "aralkylenediyl" refers to an alkylenediyl" substituted with one or more aryl groups.
The term "peptide bond" is used in its common accepted meaning.
The term "hydrolytically-labile bond" refers to a bond that is capable of undergoing hydrolysis, such as an ester, amide, acetal, or hydrazone bond. Preferably, the hydrolytically-labile bond is labile under acid conditions.
The term "halo" refers to chloro, bromo, iodo and fluoro atoms.
The term "saccharide" is used in its common accepted meaning. The terms "polysaccharide" and "oligosaccharide" refer to carbohydrate molecules containing more than one saccharide unit.
The term "activating/protecting group" refers to a group in a multifunctional compound which may temporarily activate or temporarily block a reactive site wherein a chemical reaction is to be carried out selectively at a reactive site. The reactive site may be other than the site occupied by the "activating/protecting group." The activating/protecting groups referred to, in the context of the present invention, are those commonly known activating/protecting groups including, but not limited to, activating groups such as N-succinimidyl, pentachlorophenyl, pentafluorophenyl, para-nitrophenyl, dinitrophenyl, N-phthalimido, N-norbornyl, cyanomethyl, pyridyl, trichlorotriazine, 5-chloroquinilino, and protecting groups such as N-(9-fluorenylmethoxycarbonyl) (Fmoc), carbobenzyloxy (Cbz), 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde) and imidazolyl.
The term "prepolymer" refers to a reactant used to make a polymer, that is, to monomers and other subunits from which polymers may be formed.
The term "therapeutically effective amount" refers to the amount which, when administered to a patient for treating a disease, is sufficient to effect such treatment for the disease.
The term "treating" or "treatment" is intended to include inhibiting the disease (i.e., arresting its development) and relieving the disease (i.e., causing regression of the disease).
I. The Degradable Polymers The novel degradable polymers are represented by Formula (I) R R~
~Xw ~ iY CI) ~~O O ~~O n wherein R and R1 are selected from the group consisting of hydrogen, C1_lg alkyl, Cz_ is alkenyl, Cz_l8 alkynyl, C6_i$ aryl, C~-18 alkaryl and C~_1$ aralkyl groups;
X is a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1_zoo a~anediyl, Cz_zoo alkenediyl, Cz-zoo alkynediyl, C6_zoo cycloallcanediyl, C6_zoo cycloalkenediyl, C6_zoo cycloalkynediyl, C6_zoo arylenediyl, C6_zoo alkarylenediyl, C6_zoo aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2-10,000.
I. Presently Preferred Embodiments In a preferred embodiment, R and Rl in Formula (I) are the same and are selected from hydrogen, Cl_lz alkyl, Cz_lz alkenyl, Cz_lz alkynyl, C6_iz aryl, C~_lz alkaryl and C7_iz aralkyl groups; more preferably hydrogen, CI_4 alkyl, Cz_4 alkenyl, Cz_4 alkynyl, C6_lo aryl, C~_ to alkaryl and C~_lo aralkyl groups; most preferably hydrogen, Cl_4 alkyl, and Cz_4 alkenyl.
In a preferred embodiment X is a Cl_z4 alkanediyl group, where the alkanediyl group is optionally substituted within the carbon backbone with one or more groups selected from C(O), C(O)NRI, C(O)O, >NRz', wherein N is bound to two carbon atoms within the carbon backbone and Rz' is hydrogen, or is a group capable of displacement so that N
is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group may comprise a pendant group or groups selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups.
In an embodiment of the invention, the alkanediyl group or the pendant group or groups comprises a primary, secondary, or tertiary amine group.
Most preferably, X does not comprise a saccharide, oligosaccharide or polysaccharide.
In the definition of X, any alkyl group or moiety is preferably C1_1$ alkyl, any alkenyl group or moiety is preferably Cz_l8 alkenyl, any alkynyl group or moiety is preferably Cz_lz alkynyl, any aryl group or moiety is preferably C6_z4 aryl, any alkaryl group or moiety is preferably C~_z4 alkaryl and any aralkyl group or moiety is preferably C~_z4 arallcyl, any cycloalkyl group or moiety is preferably C4_za cycloalkyl, any cycloalkenyl group or moiety is preferably CS_z4 cycloalkenyl, and any cycloalkynyl group or moiety is preferably CS_z4 cycloalkynyl.
Most preferably X is selected from the groups (1V)-(VIII) below:
a a 3 R2w ~ N R3 ~I~ Rz/ \~~~~0/ R M
O R~z ~O/ O Rs a a R ~N ~O N ~R3 N9 Rz/N~lt~~~N~R3 Nil) H ''2 ~H O Rs H
R m O Rs O
R \O a'I~"~~m Ra (VIII) O O
wherein R2 and R3 are independently selected from covalent bonds or C1_l8 alkanediyl groups terminating in OH or vinyl ether;
each R12 is independently selected from synthetic or natural amino acid side chains;
each R4 is independently selected from the group consisting of hydrogen, activating/protecting groups and the groups (IX), (X), (XI) and (XII) O R'3 ~(~
~NH ~ ~~NH
~ ~Im m O R~s R~2 H O R~
~~NH ~ ~~NH PaO
m0 R~s m ~~ ~H
R~2 RS and R6 are selected from the group consisting of -NH2~ -NHR13, -OR13, wherein each R13 is independently selected from hydrogen, Cl_4 alkyl, and activating/protecting groups; and m is an integer of 0-20.
Y preferably is represented by the formula -(C"Ha"O)qC"H2", wherein n is an integer of 2-10, preferably 2 or 3 and q is an integer of 1 to 200.
The molecular weight of the polymer of Formula (I) is preferably in the range of 10,000-100,000.
II. Preparation of the Polymer of Formula (I) The polymer of Formula I
R R' 0~ ~Xw ~ iY (I) ~~O O ~~O n wherein R and Rl are selected from the group consisting of hydrogen, Cl_l8 alkyl, Cz_ 1$ alkenyl, C2_1$ alkynyl, C6_l8 aryl, C~_1$ alkaryl and C~_ls aralkyl groups;
X is a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of Cl_zoo alkanediyl, Cz_zoo alkenediyl, Cz-zoo a~ynediyl, C6_zoo cycloalkanediyl, C6_2oo cycloallcenediyl, Cs-zoo cycloalkynediyl, C6_zoo ~ylenediyl, C6_zoo alkarylenediyl, C6_zoo ~'a~Ylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2-10,000 may be prepared by the reaction of a diol of Formula (II) HO~Y~OH (II) with a divinyl ether of Formula (III) R R~ (III) ~O~ X ~p~
wherein R, Rl, X and Y are as defined above.
The diol of Formula (II) preferably is a polyethylene glycol or polypropylene glycol compound having a molecular weight in the range 100-20,000, more preferably polyethylene glycol having a molecular weight in the range 200-10,000, most preferably polyethylene glycol having a molecular weight in the range 200-5,000, in particular a molecular weight of approximately 200-4,000. Such materials are widely available from such commercial sources as Sigma-Aldrich Corporation (St. Louis, MO) and Shearwater Polymers. Inc.
(Huntsville, AL). It will be understood by one of ordinary skill in the art that the reactant of Formula (II) may also comprise any diol of Formula (II), such as other glycols and diols suitable for use in biomaterials.
The divinyl ether of Formula (III) may be obtained commercially or may be made by any suitable means known in the art. For example, commercially-obtained amino vinyl ether may be combined with methyl esters to provide the divinyl ethers of Formula (III). Similarly, the hydroxy vinyl ether compound is commercially available, and may be used to make polyacetal polymers with ester moieties in the main chain. The methyl esters may comprise, for example, esters such as malonates, imines such as iminodiacetates, and other compounds known in the art. Symmetric, achiral methyl esters are preferred synthetic precursors.
The polymerization reaction may be carried out in a solventless system, although preferably the reaction takes place in the presence of an organic solvent selected from aliphatic or aromatic hydrocarbons, which may be optionally halogenated, ethers (including cyclic ethers), dialkylsulfoxides and alcohols (preferably sterically hindered alcohols, for example secondary or tertiary alcohols). Preferred solvents include tetrahydrofuran (THF), dichloromethane, and toluene. A particularly preferred solvent is toluene.
The polymerization is generally carried out in the presence of a suitable catalyst such as a catalyst for acid-catalysis, for example, hydrochloric acid, sulfuric acid, phosphoric acid, p-toluenesulfonic acid, methanesulfonic acid, acetic acid, n-butyric acid, trifluoroacetic acid or oxalic acid. A preferred catalyst is p-toluene sulfonic acid (p-TSA).
The polymerization is conducted at a temperature of -10 °C-200 °C, preferably 20 °C -120 °C, most preferably between about 25 °C and 60 °C.
I. Polyacetal Conjugates of Bioactive Agents and Their Preparation The functionalized polymers of the invention comprise polymers with bioactive functionality, and thus comprise polymers that include bioactive agents. The degradable polyacetal polymers of the invention comprising bioactive functionality may be formed from substrates that include bioactive agents, from polymers to which bioactive agents are conjugated, and from substrates that combine to form bioactive agents.
A bioactive agent may be attached to X in Formula (I). In a particularly preferred embodiment, when X is (IX), (X), (XI), (XII), or >NR2~, wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, the N atoms of the groups >NR2~, (IX), (X), (XI) and (XII) are, or are capable of being covalently attached to the bioactive agent.
The bioactive agent preferably is a pharmaceutically active agent (a "drug").
Suitable drugs include any drugs for which prolonged action and/or a targeted intracellular delivery is desirable, and include anticancer agents, for example doxorubicin, daunomycin, paclitaxel, taxotere, and the like, most preferably, doxorubicin. Other bioactive agents include polypeptides and proteins. The method of attachment may vary somewhat according to the bioactive agent, as is described below; and a person of ordinary skill in the art will be able, having selected a desired bioactive agent, using their knowledge and the disclosure of this application, to attach the bioactive agent to a degradable polyacetal polymer of this invention, thereby forming a conjugated polymer of this invention.
The attachment of the bioactive agent to the polymer of Formula (I) may be effected by the reaction of the polymer with the bioactive agent or a bioactive agent precursor.
Bioactive agents may be attached to the polymer in any suitable manner.
Preferably the attachment is effected subsequent to the polymerization reaction to produce the degradable polymer of Formula (1).
Attachment of bioactive agents may be effected in other ways as well. For example, attachment may be by linkages comprised of groups that covalently couple and cross-link the agents to the polymers. Such linkages may comprise disulfide linkages or ester bonds, or may be acid-labile linkages such as hydrazone linkage as described by Greenfield et al., Gazzcez"
Res., 50, 6600-6607 (1990), and references therein. Alternatively, the attachment may be via a "protected" disulfide bond that sterically inhibits attack from thiolate ions, such as are found in the coupling agents S-4-succinimidyloxycarbonyl-a-methyl benzyl thiosulfate (SMBT) S and 4-succinimidyloxycarbonyl-.alpha.-methyl-a-(2-pyridyldithio) toluene (SMPT), in the manner disclosed in U.S. Patent No. 6,048,736 to Kosak.
Where the bioactive agent has an amino group, it may be useful to form a reactive carbonate half ester in the polymer, P, P-O-CO-X, wherein X is a good leaving group, using reagents such as carbonyl diimidazole, p-nitrophenyl chloroformate or bis-N-succinimidyl carbonate. The activated polymer P, P-O-CO-X, may then be reacted with the bioactive agent under conditions which do not destroy its activity, leading predominantly to urethane linkages attached through the amino group. For example, carbonyl diimida.zole, can be reacted with terminal hydroxyl groups of the polymer. The reaction mixture may be quenched in aqueous solution at neutral pH and the activated polymer isolated, for example by dialysis or size exclusion chromatography, as disclosed in U.S. Patent No. 5, 468,478 to Saifer et al.
Attachment (conjugation) of bioactive agents may be effected by reaction with polymers or monomers with electrophilic functionality. For example, prepolyrners comprising electrophilic pendant chain functionalized monomers which comprise, for example, diols or bis-vinyl ethers, may be used for this purpose.
The following reaction scheme illustrates one route to the production of a polymer-doxorubicin drug conjugate:
~O~O O~O
O NH
O HO-PEG-OH
o,~
~J0 Scheme 2 In this scheme, PEG is the residue of a polyethylene glycol (without the terminal hydroxy groups), the terminal OH groups of the polyethylene glycol being explicitly shown when the entire glycol is meant.
Bioactive agents that may be attached (conjugated) to the polyacetal polymers of the invention include polypeptides and proteins. Such conjugation may be effected at pendant chains and at terminal groups. For example, conjugated polyacetal polymers of the invention include proteins conjugated with a degradable polyacetal polymer.
I. Administration and Pharmaceutical Composition Compositions comprising the degradable polyacetal polymers, with or without attached bioactive agents, are water-soluble or colloidal suspension compositions suitable for incorporation into pharmaceutical solutions or pharmaceutical compositions, or for delivery to an animal or patient for treatment. For example, polyacetal polymers of the invention are Ho.,.. I , HO O OH O
soluble in water and water solutions, such as saline, phosphate buffered saline (PBS), and other buffered solutions. The polyacetal polymers are soluble in solutions of widely varying pH.
In general, degradable polyacetal polymer compositions will be administered in therapeutically effective amounts by any of the usual modes known in the art.
Degradable polyacetal polymers with attached bioactive agents may be directly delivered to solutions bathing cells, tissues or organs in vitro. Pharmaceutical compositions comprising bioactive agents attached to degradable polyacetal polymers may be administered to an animal, including a human, by one of the following routes: oral, topical, systemic (e.g. transdennal, intranasal, or by suppository), or parenteral (e.g. intramuscular, subcutaneous, or intravenous injection). Compositions may take the form of tablets, pills, capsules, semisolids, powders, sustained release formulations, solutions, suspensions, elixirs, aerosols, or any other appropriate compositions; and comprise a bioactive agent attached to a degradable polyacetal polymer of the invention in combination with at least one pharmaceutically acceptable excipient. Suitable excipients are well known to persons of ordinary skill in the art, and they, and the methods of formulating the compositions, may be found in such standard references as Alfonso AR: Remivcgto~'s Pharmaceutical Sciehees, I7th ed., Mack Publishing Company, Easton PA, 1985. Suitable liquid carriers, especially for injectable solutions, include water, aqueous saline solution, aqueous dextrose solution, and glycols.
Pharmaceutical formulations comprising bioactive agents attached to degradable polyacetal polymers of the invention can be prepared according to any method known to the art for the manufacture of pharmaceuticals. Such formulations can contain sweetening agents, flavoring agents, coloring agents and preserving agents. Any such formulation can be admixed with nontoxic pharmaceutically acceptable excipients which are suitable for manufacture.
Pharmaceutical formulations for oral administration can be formulated using pharmaceutically acceptable carriers well known in the art in dosages suitable for oral administration. Such carriers enable the pharmaceutical formulations to be formulated in unit dosage forms as tablets, pills, powder, dragees, capsules, liquids, lozenges, gels, syrups, slurries, suspensions, etc. suitable for ingestion by the patient.
Intravenous injectable compositions are comprised of a polymer-drug conjugate of the invention in combination with at least one pharmaceutically acceptable liquid carrier.
Acceptable liquid carriers are non-toxic, aid administration, and do not adversely affect the therapeutic benefit of the polymer-drug conjugate. Such suitable carriers include, but axe not limited to, water, saline, aqueous dextrose and glycols. Further, excipients and other agents may be included in pharmaceutical compositions along with the degradable polyacetal polymers and attached bioactive agents. In addition, other additives and agents, such as antioxidants, antiseptic or antibiotic agents, buffers, stabilizers, solubilizers and other agents, may be added to degradable polyacetal polymer compositions of the invention.
For example, dimethylsulfoxide, benzoic acid, ascorbic acid, or tocopherol may be included in pharmaceutical compositions comprising degradable polyacetal polymer compositions of the invention. Thus, injectable compositions comprising bioactive agents conjugated to degradable polyacetal polymers will preferably comprise water or saline solutions or emulsions, pharmaceutically acceptable Garners, and may further comprise buffering agents, such as phosphate buffer or HEPES buffer, and optionally other agents.
In general, the polymer-drug conjugates of the invention will be administered in therapeutically effective amounts via intravenous injection. A therapeutically effective amount may vary depending on the severity of the disease, the age and relative health of the subject, the potency of the conjugate used and other factors. A
therapeutically effective amount may range from about 0.001 milligram per Kg (mg/Kg) body weight per day to 100 mg/Kg body weight per day. Preferably the amount will be about 0.1 to 10 mg/Kg/day.
Therefore, a therapeutically effective amount for a 70 Kg patient may range from about 0.07 to 7000 mg/day, preferably about 7 to 700 mg/day. A person of ordinary skill in the art of treating diseases such as cancer will, without undue experimentation, having regard to that skill and this disclosure, be able to determine a therapeutically effective amount of a particular bioactive agent attached to a degradable polyacetal polymer for practice of tlus invention.
The degradable polyacetal polymers of the invention, with or without attached bioactive agents, may be dried or lyophilized and stored in that condition for a considerable length of time without significant degradation or decomposition. Such dried or lyophilized compositions may be reconstituted for use, e.g., for injection, at a convenient time after storage by addition of an appropriate amount of a suitable liquid, preferably a buffered water solution, such as saline. An appropriate amount is that amount sufficient to provide the desired volume so as to result in a solution of the desired final concentration.
Excipients useful for preparation of lyophilized or freeze-dried compositions include saccharides, amino acids, and salts such as inorganic salts. Saccharides may be, for example, monosaccharides, such as glucose and fructose, disaccharides such as maltose, lactose, and sucrose, polysaccharides such as dextran and starch, and sugax alcohols, such as mannitol sorbitol and glycerol. Amino acids may include, for example, glycine, and salts may include, for example, sodium chloride and potassium chloride. Such excipients may be used alone or in combination, and may be useful for inhibiting aggregation in the reconstituted polymer solution.
The amount of a polymer-drug conjugate of the invention in the composition may vary. In general, the final composition will comprise from about 0.001% w/w to 30 % w/w of the polymer-drug conjugate, preferably about 0.01 % w/w to 10% w/w, more preferably about 0.1 % w/w to 5 % w/w with the remainder being the caxrier or carriers.
I. Pharmacology and Utility Degradable polymers are useful in a wide variety of pharmaceutical and biomedical applications. Uses for degradable polymers include coatings for drug tablets, contact lens coatings, coatings for surgical implants and medical devices, gels, as ingredients in topical and optical pharmaceutical solutions, in pharmaceutical formulations including delayed release pharmaceutical formulations and in targeted drug formulations.
Conjugation of bioactive agents, such as anticancer drugs, with degradable polymers helps to enhance the efficacy of the bioactive agent.
Degradable polyacetal polymers of the invention are suitable for use in pharmaceutical and biomedical applications with superior properties compared to prior materials. The polyacetal polymers of the invention are degradable under physiological conditions on a time-scale suitable fox effective delivery of bioactive agents in an animal. In addition, the biodistribution of degradable polyacetal polymers of the invention within the body and bloodstream of the animal receiving the polymer is favorable for the effective delivery of bioactive agents for the treatment of many diseases. The polymers and bioactive agents remain in the bloodstream for hours, not minutes, do not preferentially go to the liver, but remain in circulation so as to provide for the prolonged action of the bioactive agents, and are not toxic.
The stability of the degradable polyacetals of the invention differs in solutions of different pH. Degradable polyacetals of the invention are quite stable in water solutions near neutral pH, less so in more acidic solutions. As shown in Figure 1, polyacetal 3 (at pH 7 and at 37 °C) was quite stable, with little change in the molecular weight at time points ranging from 1 to 505 h (21 days). However, polyacetal 3 was less stable when dissolved at pH 5.5 at 37 °C. As shown in Figure 2, polyacetal 3 had essentially degraded to the PEG monomeric units by 155 h (6.5 days). The results of three such stability experiments are shown in Figure 3, in which the degradation of polyacetal 3 over time is shown as the percent molecular weight loss (Mw) at each pH value vs. time for the pH 5.5, 6.5 and 7.4.
The polyacetal polymers of the invention axe not toxic to cells. This is shown in Figure 4, which present the results of a red blood cell (RBC) lysis assay of amino polyacetal 3. No RBC lysis was observed in this assay for polyacetal 3. Dextran was the control which did not display RBC lysis, while polyethylene imine) (PEI) was the control which caused RBC lysis. In addition, direct measurements of cell toxicity on cells in culture resulted in no measured cytotoxicity. As shown in Figure 5, the cytotoxicity of polyacetal 3 was measured using the B16F10 cell line. Polyacetal 3 did not display cytoxicity in this assay compared to polylysine which was used as a cytotoxic control. Dextran was used as a noncytotoxic control.
Similar to the results shown with polyacetal polymer 3 polyacetal polymer 22 is also sensitive to pH. This is shown in Figure 6, in which are shown superimposed GPC traces of amino polyacetal 22 from a phosphate buffered solution at pH 7.4 and a solution where the pH was adjusted to pH 1-2 by addition of HCI. The polyacetal 22 completely degraded within minutes upon exposure at pH 1-2 to give the trace shown by the arrow labeled pH 1-2. This GPC trace is consistent with the molecular weight of PEG3,4oo. The degradation profile of amino polyacetal 22 at pH 7.5 and pH 5.5 is shown in Figure 7, where the loss in molecular weight is shown as a function of time. The degradation study shown in Figure 7 was conducted at 37 °C and at a concentration of 3 mg/ml of polyacetal 22;
three separate samples were analyzed at each pH. Polyacetal 22 degrades more rapidly in the mildly acidic medium at pH 5.5 than in the relatively neutral physiological pH 7.4. Lysosomal pH is in the range of pH 5.0 to pH 5.5. The experimental pH value of 5.5 was selected to match the Iysosomal pH, which would be that encountered by a physiologically soluble polymer conjugate upon cellular uptake by endocytosis within the lysosome.
The in vitro biocompatibility of amino polyacetal 22 is shown in Figures 8a, 8b, and 9. Figure 8a shows the results of red blood cell lysis experiments at 1 hour, and Figure 8b shows the results of red blood cell lysis experiments at 5 hours. Figure 9 shows the results of cytotoxicity experiments. These experiments show that polyacetal 22 is not lytic or toxic to these cells, and so has a favorable biocompatibility profile.
Polyacetal polymers of the invention do not cause lysis of red blood cells.
The results of a red blood cell (RBC) lysis assay of amino polyacetal 22 and its degradation products is shown over a 1 hour time period in Figure 8a. The degradation products were obtained by dissolving polyacetal 22 in phosphate buffered saline (PBS), adjusting the pH
to 1-2 by the addition of HCl to allow the polyacetal to degrade, then adding a small amount of NaOH to readjust the pH to 7.4. No RBC lysis was observed in this assay for polyacetal 22. Dextran was used as a control; it did not display RBC lysis. Polyethylene imine) (PEI) was also used as a control; it caused RBC lysis. Similarly, in Figure 8b, the results of a red blood cell lysis assay of amino polyacetal 22 and its degradation products are shown over a 5 hour time period. No RBC lysis was observed in this assay for polyacetal 22. Dextran and polyethylene imine) (PEI) were used as control compounds again: dextran did not display RBC
lysis, while PEI did cause RBC lysis. Thus, the results shown in Figures 8a and 8b demonstrate that the polyacetal polymer of the invention, polyacetal 22, was not lytic for red blood cells for up to five hours.
In addition, polyacetal polymers of the invention are not cytotoxic. A
cytotoxicity assay of amine pendant chain polyacetal 22 using B16F10 cell Iine is shown in Figure 9. As shown in Figure 9, polylysine is cytotoxic at concentrations well below 0.1 mg/ml. However, polyacetal 22 did not display cytoxicity in this assay at concentrations of up to several mg/ml, and thus is extremely well tolerated by these cells as compared to the cytotoxic control compound, polylysine. Dextran, used as a noncytotoxic control, was also found not to be cytotoxic even at relatively high concentrations.
Polyacetals of the invention remain in circulation in the blood with relatively little loss from the blood circulation to the organs. lzsl labeled polyacetal polymers of the invention may be formed using Bolton-Hunter methods, as shown in Figures 10 and 11 and described in Example 7. In the body distribution study shown in Figure 12, polyacetal 23 was predominantly in the blood, at both at 5 min and at 1 hour after administration. The continued presence of polyacetal 23 in the blood in significant amounts at one hour is a surprising and favorable property of the polyacetal of the invention. This study shows that polyacetal 23 remains in the blood without accumulating in the organs shown. In particular, and in contrast to many previously-known polymers, the polyacetal of the invention is not significantly taken up by the liver but remains substantially in circulation in the blood for an hour. Thus, the polyacetal of the invention possesses the favorable properties of long-duration presence in the blood, of very little removal from circulation by the organs, and, since very little of the polyacetal is lost to the liver, relatively little degradation by the liver.
Functionalized polymers of the invention, such as may be formed by synthesis from fwctionalized precursors or by attaclnnent of bioactiee agents, such as anticancer drugs, to degradable polyacetal polymers of the invention may be effective to enhance the efficacy of the bioactive agent. As shown in Figure 12, polyacetal polymers remain in circulation with relatively little removal or loss from the blood on a timescale of hours, a property which enhances the effectiveness of anticancer drugs attached to degradable polyacetal polymers of the invention. Higher dosages of the anticancer drugs, which are more effective in treating the cancerous tissue than lower dosages, are tolerated by animals when anticancer drugs are attached to degradable polyacetal polymers of the invention.
I. Prepolymers of Formula (XIII) The prepolymers are novel divinylethers represented by Formula (XIII) ' (X111) wherein R' and R8 are selected from the same groups as R and Rl, Z is a C1_z4 alkanediyl group, optionally substituted within the carbon backbone with one or more or a mixture of the groups selected from carbonyl, peptide, ester, >NRz', wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group comprises a pendant group R9, wherein R9, is selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloallcenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups.
In a preferred embodiment, Rg contains at least one activating/protecting group.
In the definition of Z, any alkyl group or moiety is preferably Cl_l8 alkyl, any alkenyl group or moiety is preferably C2_lg alkenyl, any alkynyl group or moiety is preferably C2_lz alkynyl, any aryl group or moiety is preferably C6_24 aryl, any alkaryl group or moiety is preferably C~_Z4 alkaryl and any aralkyl group or moiety is preferably C~_24 aralkyl, any cycloalkyl group or moiety is preferably C4_z4 cycloalkyl, any cycloalkenyl group or moiety is preferably CS_24 cycloalkenyl, and any cycloalkynyl group or moiety is preferably CS_24 cycloalkynyl. Symmetric achiral methyl esters are preferred synthetic precursors.
Preferably Z has a structure (X1V), (XV), (XVI), (XVII) and (XVIII), below:
OI1 R9 O L. O
R' ~O~j~ ~Rm (XIV) Rz/0~~~~~O~Rm lRY~z ~O IO Y R~ a O R9 ' O Ii O
R~ N /R~~ ~1) RzrN~\~~~HrRt~ ~If) N~ ~ Ip1'1 ~,R~ '9 O Rs O
R~ ~O'~u~~0 O m X111) P O P P P R
O
wherein Rl° and Rll are selected from covalent bonds, and Cl_6 allcanediyl groups;
R9 is as defined above, R12 are selected from synthetic or natural amino acid side chains; and p is an integer of 0-20.
Prepolymers as disclosed herein may be formed by any suitable method, including such methods as are disclosed in the Examples, such as Examples 3, 4 and 5 following.
A process for the preparation of a prepolyrner of Formula (XII~ may comprise the following steps. To prepare a prepolymer of Formula (XIII):
R' R9 ~ (X111) ~Oit~O
wherein R~ and R8 are selected from the group consisting of hydrogen, C1_l8 alkyl, CZ_la alkenyl, CZ_l8 alkynyl, C6_l8 a aryl, C~_l8 alkaryl and C~_l8 aralkyl groups;
Z is a Cl_z4 alkanediyl group, where the alkanediyl group is optionally substituted within the carbon backbone with one or more groups selected from C(O), C(O)NRI, C(O)O, >NRZ~, wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group comprises a pendant group R9 selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoall~yl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups;
the steps of the process comprise:
reacting a methyl ester of Formula (XIX) O (XIX) H3C0 X' \OCH3 with a vinylether of Formula (XX) H2N ~YwO~ (~) or Formula (XXI) H O~YwO~ (XXI ) wherein X comprises a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond, and Y is a group selected from the group consisting of linear and branched C1_zoo alkanediyl, Cz_zoo alkenediyl, Cz_zoo a~ynediyl, C6_zoo cycloalkanediyl, C6_zoo cycloallcenediyl, C6_zoo cycloalkynediyl, C6_zoo az'Ylenediyl, C~_zoo alkarylenediyl, C~_zoo. aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone.
In a preferred embodiment of the method of preparing a prepolyrner of Formula (XIII) R9 contains at least one activating/protecting group.
These prepolymers are particularly useful for the preparation of the polymers of Formula (I) by methods known in the art and as illustrated in the Examples.
Functionalized prepolymers, such as prepolymers functionalized with bioactive agents or precursors to bioactive agents, are useful for the preparation of polymers of formula comprising bioactive agents. The following structures represent some particularly preferred prepolymers of the present invention.
O
N O O~OW% ~'O
O O
~O
~~''O
O O HN O
'~ ~,O O p O
O ~ ~ ~ U
O O
~O~N ~~0~
H
NHFmoc O O
~O~~N N~O~
H H
NH
O
Cbb-I ),N
NHFmoc O
O ~ O
~O~N~N~N~O~
H , H
Scheme 1 EXAMPLES
General The degradable polymers of the present invention may be prepared by the reaction of polyethylene glycol) (PEG) as the source of diol (PEG' S with molecular weights of 3,400 g/mol were used) and commercially available triethylene glycol di-vinyl ether.
PEG is selected as the diol because it is generally recognized as safe (GR.AS) by drug regulatory authorities and is widely used in pharmaceutical formulation. However, it will be appreciated by those of ordinary skill in the art that other diols, including PEGs of lower or higher molecular weight, are also suitable for the practice of the invention. The use of the tmfunctionalized divinyl ether, triethylene glycol di-vinyl ether, in the preliminary experiments was conducted to confirm a suitable degradation profile (needed for lysosomal degradation) and to confirm ih vitro biocompatibility. Tt will be understood by one of ordinary skill in the art that degradable polyacetal polymers of the invention may also be prepared from functionalized starting materials. For example, functionalized vinyl ethers, particularly functionalized divinyl ethers, may be used as starting materials in the preparation of the degradable polyacetal polymers of the invention. Each following experimental example is preceded by a scheme summarizing the reaction involved. In each case m is an integer representing a PEG molecule of the identified molecular weight Mn.
Example 1. Synthesis of polyacetal 3 by polymerization in toluene HO~O~H + ~O~O~O
ll '''' J m _1 2 ~, pTSA (cat)/toluene O O~ O~O~O~O~O
-m1i n Polyethylene glycol) (Mn = 3,400 g/mol, 17.0 g, 5.0 mmol, 1.0 ec~, para-toluenesulfonic acid monohydrate (0.03 g, 0. 15 mmol, 0.03 ec~ and toluene (60 ml) were added to a 100 ml round bottom flask which was equipped with a stirring bar and fitted with a thermometer, Dean Starlc trap and condenser. An azeotropic distillation of the stirred toluene solution (oil bath, T=150 °C) under nitrogen proceeded for two hours.
The solution was then allowed to cool to ~ 50 °C and tri(ethylene glycol) divinyl ether (1.073 g, 1.083 ml, 5.2 mmol, 1.04 eq) was added by syringe. Within one minute the reaction mixture became visibly more viscous and after 15 minutes the viscosity appeared to be very high.
Toluene (30.0 ml) was added to decrease the viscosity and the clear colorless reaction mixture was stirred a further 2 hours at ambient temperature. Aqueous NaHC03 (8.0%, 2.0 ml) was added to the reaction mixture which was then rapidly stirred for 15 minutes. The aqueous phase was allowed to settle and the toluene phase was carefully decanted into stirred hexane (200 ml) to precipitate the polyacetal. After stirring in the hexane for an additional 10 minutes the polyacetal was collected and placed into a fresh solution of hexane and stirred for a further 10 minutes. The polyacetal was again collected and then dried in vacuum at 50 °C for 4 hours to give a white fluffy solid. The molecular weight was determined to be Mw=42,806 g/mol, Mn=26760 g/mol; polydispersity-1.60 by GPC. The GPC was calibrated with PEG
standards;
56,000, 23,500 and 5598 g/mol.
Example 2. Synthesis of polyacetal 3 by polymerization in THF
A suspension of PEG3aoo (1.041 g, 0.306 mmol) and para-toluenesulfonic acid (25 mg) in toluene (50 ml) was heated to reflux; the flask was fitted with a Dean and Stark trap and a balloon of argon was fitted to the condenser. After 150 min most of the toluene was distilled off. To the residue was added the divinyl ether (0.306 mmol, 1.0 eq) in freshly distilled THF (10 ml). The mixture was stirred at room temperature under argon for 16h.
Triethylamine (0.2 ml) was added and the mixture was stirred for 5 minutes.
The mixture was poured into hexane (300 ml) with rapid stirring, after 5 min the hexane was decanted off and the residue was washed with further hexane (200 ml) for 30 minutes. The polymer was filtered off.
This same procedure was used for polymerizations conducted in dichloromethane.
Exam 1p a 3. Synthesis of bis-vinyl ethers useful for preparing polyacetals The vinyl ethers were made from methyl esters using the commercially available amino vinyl ether. This avoided the use of heavy metals to make the vinyl ether moiety.
~ ~ !
CH30- v 'OCH3 '~ H2N O ~ > // _O NH NH O
s A solution of 3-amino-1-propanol vinyl ether 5 (0.27 mmol, 2.2 eq) and dimethyl malonate 4 (0. 12 mmol, 1 eq) in dichloromethane (S.0 ml) was stirred at ambient temperature for 3 days. The reaction mixture was diluted with dichloromethane (SO ml) and washed with water (2 x 35 ml), conc. NaCI solution (3S ml) and dried over MgS04. The solvent was evaporated to give a semi-solid residue which was triturated with ether-hexane (1:l) to give the bis vinyl ether 6.
Exam 1p a 4. Synthesis of bis-vinyl ethers useful for preparing polyacetals with functionality to conjugate bioactive compounds H3CO~~~OCH3 + ~O~NH, > ~p~NH~~NH~O~
A saturated aqueous Na2C03 solution (20 ml) of dimethyl iminodiacetate 7 (10 mmol, 1.0 eq) and 3-amino-1-propanol vinyl ether 5 (40 mmol, 4.0 eq) was stirred for 1 h at 90 °C.
The solution was cooled and extracted three times with ethyl acetate (SO ml each time). The organic layer was washed with brine, dried over MgS04, and rotoevaporated to give the bis-vinyl ether 8 as a white crystalline solid.
The bis-vinyl ether 8 was then allowed to react with various acylating agents (e.g.
Fmoc protected glycine N-hydroxysuccinimde ester and benzyl chloroformate) to protect (block) the amino functionality in 8 prior to polymerization. The protecting (blocking) group is important because it allows polymerization to proceed without competitive side reactions with the conjugating fimctionality. After polymerization it must be removed without causing degradation of the polyacetal. This strategy is illustrated in Example 5 for the preparation and polymerization of glutamic acid derived bis-vinyl ether 11 and the subsequent deprotection of the polyacetal.
Exam 1p a 5. Synthesis of bis-vinyl ethers useful for preparing polyacetals with a protected primary amine functionality ~~11). Preparation of a amino pendant chain functionalized polyacetal using a pendant chain functionalized bis-vinyl ether monomer (11-X13) Synthesis of Fmoc-glutamyl chloride 10 ~9-X10) \ ~ ~,/~~ ~~2 HO~~~OH > CI~~~~~CI 5 ~O~NH NH~O
INHFmoc INHFmoc NHFmoc _9 _10 _11 To a suspension of Fmoc-glutamic acid 9 (3.13g, 8.5 mmol) in anhydrous CH2C12 (50 ml) was added oxalyl chloride (5.0g, 39 mmol). The mixture was cooled to 0 °C and DMF (2 drops) was added. The mixture was stirred at 0 °C for 1 h then at ambient temperature for 1 h under argon atmosphere. Freshly distilled THF (6.0 rnl) was added and the mixture was stirred at room temperature for 1 h under argon. Hexane (400 ml) was added and the mixture stirred at ambient temperature for 30 minutes. The hexane was decanted off and the residue recrystallized from CHaCl2/hexane to give 10 as white crystals (1.41g).
Synthesis of Fmoc glutamic acid divinyl ether 11 10-X11 To the bis-acid chloride 10 (1.41 g, 3.47 mmol) in anhydrous CH2C12 (25 ml) was added a solution of 3-aminopropyl vinyl ether 5 (701 mg, 6.94 mmol) and NaHC03 in water (10 ml) dropwise over 5 min with vigorous stirring at 0 °C. The mixture was stirred at 0 °C
for 10 min then at ambient temperature for 1 hour. The mixture was diluted with CHaCIa (200 ml) then washed with 2% aqueous NaHC03 (150 ml), brine (150 ml) and then dried with MgS04. Evaporation gave a pale brown solid which was recrystallized from isopropanol/hexane to yield the divinyl ether 11 as an off white solid (910 mg).
Synthesis of amino-functionalized bis-vinyl ether monomer 13 (1113) O O HO~O~H
I. '" J m o 0 G~~r~~r~~c~ 1 _ NHFmoc ~ ~~~NH~NH~
_i l m-1 NHFmoc m-1 NHz A suspension of PEG34oo 1 (1.041 g, 0.306 mmol) and p-toluene sulfonic acid (25 mg) in toluene (50 ml) was heated to reflux in a round bottom flask fitted with a Dean and Stark trap to collect the water. After no further increase in water collection was observed, most of the toluene was distilled from the round bottom flask. To the residue was added the divinyl ether 11 (164 mg, 0.306 mmol) in freshly distilled (sodium-benzophenone) THF
(10 ml). The mixture was stirred at room temperature under argon for 16 h. Triethylamine (0.2 ml) was added and the mixture was stirred for 5 minutes. The mixture was poured into hexane (300 ml) with rapid stirring to precipitate the polyacetal 12. After 5 min the hexane was decanted off and the polyacetal 12 was stirred with fresh hexane (200 ml) for 30 min.
The polyacetal 12 was filtered, collected and dried in vacuum. The weight average molecular weight as determined by GPC (eluent: water, 1 mI/min; PEG standards) was 14300 g/mol.
The Fmoc group was removed by dissolving the polymer into an dichloromethane (7% weight percent) followed by the addition of morpholine and the reaction stirred for 15 min at ambient temperature. The amino functionalized polyacetal 13 was precipitated into a stirred solution of hexane (200 ml).
Example 6. Synthesis of a symmetric, achiral bis-vinyl ether 16 with a protected primary amine useful for preparing polyacetals with functionality to conjugate bioactive compounds ~ ~ ~~NHZ
H3C0' Y'OCHg Cbz-Gly-NHS> H3CO~OCHg 5 > ~O~NH~NH"~~O~
INH3+Cl- H H
O O
14 cb~Hrr 15 cbzHrr 16 A solution of Cbz-gly-NHS (0.75 g, 2.5 mmol), dimethyl aminomalonate hydrochloride 14 (0.45 g, 2.5 mmol) and triethylamine (0.375 ml) in dichloromethane (5 ml) was stirred in a 25 ml single neck round bottomed flask at ambient temperature for 12 hours.
A white precipitate assumed to be triethylamine hydrochloride was evident. The reaction mixture was diluted in dichloromethane (80 ml) and transferred to a separatory funnel, and washed with water and brine. The organic layer was dried over MgS04 filtered and the solvent removed by rotoevaporation to give the amino acid dimethylmalonate 15 as white solid (83%). Synthesis of bis-vinyl ether 16 was completed in dichloromethane by the same process as for the preparation of bis-vinyl ether 8 (Example 4).
Exam 1p a 7. Preparation and in vitro biocompatibility evaluation of a pendant chain functionalized polyacetal 22.
This example describes the preparation of a pendant chain functionalized polyacetal.
This polyacetal 22 was prepared by a terpolymerization process using a suitably functionalized diol monomer 20 for the incorporation of the pendant chain functionality onto the polyacetal. A radiolabel agent was then conjugated to polyacetal 22 to give the labelled conjugate 23 which was used in an ih vivo biodistribution study. An iya vitf o degradation study is also described for polyacetal 22 in this example.
Synthesis of a diol 20 that is functionalized to provide the pendant chain in the final polyacetal HO~ OH ' HO~ OH
HN O
To a rapidly stirred solution of amino diol 19 (1.0 g, 10.0 mmol) and NaOH (1 M, 25 ml) that was cooled to 0-2 °C with a water ice bath was slowly added a dichloromethane (10 ml) solution of Fmoc-chloride (3.4 g, 13.1 mmol, 1.2 eq) over a 1 h period.
The solution was stirred a further 1 h at 0 °C then at ambient temperature for 4 1i. The reaction mixture was transferred to a rotoevaporator and the dichloromethane was evaporated off. To the aqueous residue was added ethyl acetate (70 ml), the solution transferred to a separatory funnel and the organic layer washed with dilute aqueous HCl solution (5 %), dilute NaHC03, brine, dried over MgS04 and rotoevaporated to give a solid which was recrystallized in chloroform to give the Fmoc protected amino diol 20.
Terpolymerization process to prepare the protected amino functionalized polyacetal 21. This is a polymeric precursor to the desired amine functionalized polyacetal 22 H~ OH
HNI O
2 +
_20 H~-PEG-OH
PEG3,4oo 1 (5.005 g, 1.47 mmol, 1.0 eq) andp-toluene sulfonic acid (0.012 g) were added to a 100 ml single neck round bottom flask equipped with a stir bar.
This mixture was heated in vacuum (0.5-1.0 torn) at a temperature of 80-90 °C (oil bath) for 3.0 h. After cooling the flask was purged with nitrogen and Fmoc serinol 20 (0.461 g, I.47 mmol 1.0 eq) and freshly distilled THF (10.0 ml; distilled from sodium-benzophenone) were added to the flask.
To this stirred solution was added a solution of tri(ethylene glycol) 2 (0.601 ml, 2.94 mmol, 2.0 eq) in THF (10.0 ml). The reaction mixture was vigorously stirred at ambient temperature for 2 hour, then triethylamine (0.3 ml) was added to deprotonate the p-toluene sulfonic acid.
The reaction mixture was slowly added to a stirred solution of hexane (I00 ml) to precipitate the polyacetal 21 which was filtered, collected and dried in vacuum at ambient temperature.
GPC analysis indicated the molecular weight was about 25,000 g/mol (eluent:
phosphate buffer solution, 1 ml%min; 2 Waters Hydrogel Columns; PEG standards). H-NMR
analysis confirmed the equivalent incorporation of the Fmoc-amino diol monomer 20 and PEG into the polyacetal 21. The removal of the Fmoc group is given below.
~G~O-PE6-(~~ i eridin ~
[, J n o-PEA
3 n 21 ' 22 A solution of polyacetal 21 (2.050 g) in 20 % piperidine in dichloromethane (10 ml) was stirred at ambient temperature. Thin layer chromatography was used to monitor the reaction (eluent: ethyl acetate). The amino functionalized polyacetal 22 was isolated by first partitioning the piperidine and an unknown amount of by-products into hexane then the dichloromethane was evaporated. The residue was dissolved in THF then this solution added to a stirred solution of hexane (100 ml) to precipitate the desired amino polyacetal ~22. GPC
analysis indicated the polymer molecular weight to be about 23,000 g/mol (eluent: phosphate buffer solution 1 ml/min; 2 Waters Hydrogel Columns; PEG standards). 1H-NMR
analysis indicated the loss of the aromatic blocking group with no reduction in the acetal functionality.
Conjugation of an electrophilic compound to polyacetal 22 (~ ~ '~~ Bolton-Hunter (~ ~ '~~
~~O-PECr-~~ R~ C 'LT( 'O-PEG-J 3 NHz JJ 3 3 _22 _23 OH
The Bolton-Hunter (N-succinimidyl 3-(4-hydroxy 5-[lzsl]iodophenyl) propionate) method may be used to iodinate peptides which do not contain a tyrosine residue or peptides whose activity is destroyed by tyrosine iodination. See, for example, A.E.
Bolton et al., Biochem. J.,133, :529-38, 1973. The conjugation of Bolton-Hunter radiolabel reagent to the amine fiuictionalized polyacetal 22 gives the conjugated polyacetal 23 which is radiolabeled with lzsI.
The amino polyacetal 22 was radiolabeled using the Bolton Hunter reagent by first dissolving polyacetal 22 (50 mg) at 10 mg/mL in borate buffer (0.1 M) at pH
BACKGROUND TO THE INVENTION
FIELD OF THE INVENTION
This invention relates to degradable polyacetal polymers and therapeutic agents derived therefrom, the production of these materials, and methods of disease treatment using them.
DESCRIPTION OF THE RELATED ART
Polymer therapeutics (R. Duncan, "Polymer therapeutics for tumor specific delivery", Chem. & Ihd., 7, 262-264, 1997) are developed for biomedical applications requiring physiologically soluble polymers; and include biologically active polymers, polymer-drug conjugates, polymer-protein conjugates, and other covalent constructs of bioactive molecules.
An exemplary class of a polymer-drug conjugate is derived from copolymers of hydroxypropyl methacrylamide (HPMA), which have been extensively studied for the conjugation of cytotoxic drugs for cancer chemotherapy (R. Duncan, "Drug-polymer conjugates: potential for improved chemotherapy", A~rti-Cancer Drugs, 3, 175-210, 1992; D.
Putnam et al., "Polymer conjugates with anticancer activity", Adv. Polym.
Sci., 122, 55-123, 1995; R. Duncan et al., "The role of polymer conjugates in the diagnosis and treatment of cancer", STP PhaYma, 6, 237-263, 1996). An HPMA copolymer conjugated to doxorubicin, known as PK-l, is currently in Phase II evaluation in the UK. PK-1 displayed reduced toxicity compared to free doxorubicin in the Phase I studies (P. Vasey et al., "Phase I clinical and pharmacokinetic study of PKI (HPMA copolymer doxorubicin): first member of a new class of chemotherapeutic agents: drug-polymer conjugates" Clih.
Cahcef° Res., 5, 83-94.
1999). The maximum tolerated dose of PK-1 was 320 mg/ma, which is 4-5 times higher than the usual clinical dose of free doxorubicin. .
The polymers used to develop polymer therapeutics may also be separately developed for other biomedical applications that require the polymer be used as a material. Thus, drug release matrices (including microparticles and nanoparticles), hydrogels (including injectable gels and viscous solutions) and hybrid systems (e.g. liposomes with conjugated polyethylene glycol) on the outer surface) and devices (including rods, pellets, capsules, films, gels) can be fabricated for tissue or site specific drug delivery. Polymers are also clinically widely used as excipients in drug formulation. Within these three broad application areas:
(1) physiologically S soluble molecules, (2) materials, and (3) excipients, biomedical polymers provide a broad technology platform for optimizing the efficacy of an active therapeutic drug.
An increasing number of physiologically soluble polymers have been used as macromolecular partners for the conjugation of bioactive molecules. Many polymers have the disadvantage of being non-degradable in the polymer backbone. For example, polyethylene glycol) (C. Monfardini et al., "Stabilization of substances in circulation", Bioconjugate Chem., 9, 418-450, 1998; S. Zalipsky, "Chemistry of polyethylene glycol conjugates with biologically active molecules", Adv. D~ugDelive~y Rev, 16,-1S7-182, 1995; C.
Delgado et al., "The uses and properties of PEG-liked proteins", C~it. Rev. They. Drug CaYrie~ Syst., B, 249-304, 1992; M.L. Nucci et al., "The therapeutic values of polyethylene glycol)-modified proteins", Adv. Drug DeliveYy Rev. 6, 133-151, 1991; A. Nathan et al., Copolymers of lysine and polyethylene glycol: A new family of functionalized drug carriers", Bioconjugate Chem.
4, S4-62, 1993) and HPMA (D. Putnam et al., "Polymer conjugates with anticancer activity", Adv. Pol~m. Sci., 122, SS-123, 1995; and R. Duncan et al., "The role of polymer conjugates in the diagnosis and treatment of cancer", STP Pha~ma, 6, 237-263, 1996) copolymers have been extensively studied for conjugation. PEG is also generally used in the pharmaceutical industry as a formulation excipient. These hydrophilic polymers are soluble in physiological media, but their main disadvantage is that the polymer mainchain does not degrade in vivo.
Thus it is not possible to prohibit accumulation of these polymers in the body. Only polymers with a molecular weight lower than the renal threshold can be used for systemic 2S administration. It is imperative that for the systemic use of non-degradable polymers such as HPMA and PEG only molecules of a molecular weight which are readily cleared be administered or else long-term deleterious accumulation in healthy tissue will invariably result (L. Seymour et al., "Effect of molecular weight (Mw) of N-(2-hydroxypropyl)methacrylamide copolymers on body distributions and rate of excretion after subcutaneous, intraperitoneal and intravenous administration to rats", J. Biomed. Mate.
Res. 21, 341-1358, 1987; P. Sclmeider et al., "A review of drug-induced lysosomal disorders of the liver in man and laboratory animals", Microscopy Res. Tech. 36, 253-275, 1997; C.
Hall et al., "Experimental hypertension elicited by injections of methyl cellulose", Experientia 17, 544-454, 1961; C. Hall et al., "Macromolecular hypertension:
hypertensive cardiovascular disease from subcutaneously administered polyvinyl alcohol", Experientia 18, 38-40, 1962).
Although some natural polymers such as polysaccharides have the advantage of being degradable in vivo, e.g. dextran, they typically lack a strict structural uniformity and have the propensity upon chemical modification (i.e. conjugation of a bioactive molecule) to become inununogenic or non-degradable (J. Vercauteren et al., "Effect of the chemical modification of dextran on the degradation by dextranases", J. Bio. Comp. Polymers 5, 4-15, 1990; W.
Shalaby et al., "Chemical modification of proteins and polysaccharides and its effect on enzyme-catalyzed degradation", in: S. Shalaby, ed. Biomedical Polymers.
Designed to-degrade systems. New York: Hanser Publishers, 1994). Other polysaccharides which have been investigated for biomedical conjugation applications include chitosan (Y.
Ohya et al., "a-1,4-Polygalactosamine immobilised 5-fluorouracils through hexamethylene spacer groups via urea bonds", J. Cont. Rel.,17, 259-266, 1991), alginate (A. Al-Shamkhani et al., "Synthesis, controlled release properties and antitumor activity of alginate cis-aconityl daunomycin conjugates", Int. J. Pharm., 122, 107-119, 1995; S. Morgan et al., "Alginates as drug carriers: covalent attachment of alginates to therapeutic agents containing primary amine groups", Int. J. Pharm., 122, 121-128, 1995), hyaluronic acid (B.
Schechter et al., "Soluble polymers as carriers of cisplatinum", J. Cont. Rel., 10, 75-87, 1989), 6-O-carboxymethyl chitan (Y. Ohya et al., "In vivo and in vitro antitumor activity of CM-Chitin immobilized doxorubicins by lysosomal digestible tetrapeptide spacer groups", J. Bioact.
Compat. Polymers,10, 223-234, 1995) and 6-O-carboxymethyl pullulan (H. Nogusa et al., "Synthesis of carboxymethylpullulan peptide doxorubicin conjugates and their properties", Claem. Pharm. Bull., 43, 1931-1936, 1995).
Other natural polymers such as proteins can also be used to conjugate a bioactive molecule. For example albumin has been investigated as a protein used to conjugate a bioactive molecule (P. Balboni et al., "Activity of albumin conjugates of 5-fluorodeoxyuridine and cytosine arabinoside on poxviruses as a lysosomotropic antiviral chemotherapy", Nature, 264, 181-183, 1976; A. Trouet et al., "A covalent linkage between daunorubicin and proteins that is stable in serum and reversible by lysosomal hydrolases as required for a lysosomotropic drug-carrier conjugate. In vitro and in vivo studies", Proc.
Natl. Acad. Sci. ZISA, 79, 626-629, 1982; F. Dosio et al., "Preparation, characterization and properties in vitro and in vivo of a paclitaxel-albumin conjugate", J. Cont.
Rel., 47(3), 293-304, 1997; T. Yasuzawa et al, "Structural determination of the conjugate of human serum albumin with a mitomycin C derivative, IOW-2149, by matrix assisted laser desorption/ionization mass spectrometry", Biocohjugate Chena., 8, 391-399, 1997; A.
blunder et al., "Antitumor activity of methotrexate-albumin conjugates in rats bearing a Walker-256 carcinoma", Int. J. Cancer, 76, 884-890, 1998). The major limitations for using a protein to conjugate a bioactive compound include the propensity for inducing immunogenicity and non-specific degradation of the protein in vivo, and denaturation and irreversible alteration of the protein during preparation of the conjugate.
Other proteins such as transferrin, which binds to the transferrin receptor and thus have the potential to undergo receptor-mediated uptake (T. Tanaka et al., "Intracellular disposition and cytotoxicity of transferrin-mitomycin C conjugate in HL60 cells as a receptor-mediated drug targeting system", Biol. Pharm. Bull., 21(2), 147-152, 1998) and various immuno-conjugates (D. Gaal et al., "Low toxicity and high antitumor activity of daunomycin by conjugation to an immunopotential amphoteric branched polypeptide", Eur. J. Cayacer, 34(1), 155-16, 1998; P.
Trail et al., "Site-directed delivery of anthracyclines for the treatment of cancer", Drug Dev.
Res. 34, 196-209, 1995; E. Eno-Amooquaye et al., "Altered biodistribution of an antibody-enzyme conjugate modified with polyethylene glycol", Br. J. Cancer, 73, 1323-1327, 1996;
P. Flanagan et al., "Evaluation of antibody-[N-(2-hydroxypropyl)methacrylamide] copolymer conjugates as targetable drug-carriers. 2. Body distribution of anti Thy-1,2 antibody, anti-transferrin receptor antibody B3/25 and transferrin conjugates in DBAZ mice and activity of conjugates containing daunomycin against L1210 leukemia in vivo", J. Cont.
Rel., l8, 25-38, 1992; C. Springer et al., "Ablation of human choriocarcinoma xenografts in nude mice by antibody-directed enzyme prodrug therapy (ADEPT) with three novel compounds", Eur. J.
CajZCer,11, 1362-1366, 1991.) also have been investigated. Monodisperse molecular weight distribution is often claimed to be a significant advantage for using proteins to conjugate drugs, but this can only be useful if a single species of the protein-drug conjugate can be reproducibly prepared on adequate scale which stable on storage. This is generally not economically or technologically possible to achieve in practice. Thus, there is a need for degradable synthetic polymers developed for biomedical application, and specifically for conjugation applications, which can address the limitations inherent in the use of natural polymers for these applications.
Synthetic polymers which have been prepared and studied that are potentially degradable include polymers derived from amino acids (e.g. poly(glutamic acid) , poly[SN-(2-hydroxyethyl)-L-glutamine), (3-poly(2-hydroxyethyl aspartamide), poly(L-glutamic acid) and polylysine). These polymers when prepared for conjugation applications that require physiological solubility do not degrade in vivo to any extent within a time period of 10-100 hours. Additionally polymers and copolymers including pseudo-poly(amino acids) (K. James et al., "Pseudo-poly(amino acids: Examples for synthetic materials derived from natural metabolites", in: K. Paxk, ed., Coht~olled Drug Deliver: Challenges ahd Strategies, Washington, DC: American Chemical Society, 389-403, 1997) and polyesters such as copolymers of polylactic and poly(glycolic acid), poly(a or b-malic acid) (K.
Abdellaoui et al., "Metabolite-derived artificial polymers designed for drug targeting, cell penetration and bioresorption", Eur. J. Pha~m. Sei., 6, 61-73, 1998; T. Ouchi et al., "Synthesis and antitumor activity of conjugates of poly (a-malic acid) and 5-fluorouracil bound via ester, amide or carbamoyl bonds", J. Coyat. Rel., 12, 143-153, 1990), and block copolymers such as PEG-lysine (A. Nathan et al., "Copolymers of lysine and polyethylene glycol: A new family of functionalized drug carriers", Biocohjugate Chern., 4, 54-62, 1993.), poly(lysine citramide) (K. Abdellaoui et al, "Metabolite-derived artificial polymers designed for drug targeting, cell penetration and bioresorption", Eu~. J. PhaYna. Sci., 6, 61-73, 1998) and amino acid-PEG
derived block copolymers (G. Kwon et al., Block copolymer micelles as long-circulating drug vehicles", Adv. Drug Del. Rev., 16, 295-309, 1995; and V. Alakhov et al., "Block copolymeric biotransport carriers as versatile vehicles for drug delivery", Exp. Opih. Invest.
DYUgs, 7(9), 1453-1473, 1998) have also been investigated for conjugation.
Acetals are well known to be hydrolytically labile under mildly acidic conditions.
Thus, biomedical polymers possessing acetal linkages in the polymer mainchain may undergo enhanced rates of hydrolysis in biological environments that are mildly acidic compared to biological environments that are at neutral or basic pH. For example, soluble polyacetals that can conjugate a bioactive molecule are expected to degrade at enhanced rates at the acetal functionality during cellular uptake because of the increase in acidity during endocytosis.
Polyacetals will also display enhanced rates of hydrolysis in acidic regions of the gastrointestinal tract. Additionally polyacetals would be expected to degrade at enhanced rates at sites of diseased tissue that are mildly acidic (e.g. solid tumors).
Preparing polyacetals can be accomplished by acetal- or transacetalization reactions which result in the formation of a low molecular weight by-product (e.g. water or an alcohol).
Complete removal of such a by-product is necessary for reproducible polymerization and to ensure the polyacetal does not degrade on storage. Usually harsh conditions are required to obtain high molecular weight polymer. If functionalized monomers relevant for biomedical applications are used, such conditions can often lead to unspecif ed chemical changes in the monomer. Polyacetals can be prepared without generation of a small molecule which requires removal by cationic ring-opening polymerization using bicyclic acetals (L.
Torres et al., "A
new polymerization system for bicyclic acetals: Toward the controlled/"living"
cationic ring-opening polymerization of 6,8-dioxabicyclo[3.2.1] octane", Mac~orraolecules, 32, 6958-6962, 1999). These reaction conditions lack versatility because they require bicyclic acetal monomers that are difficult to prepare with a wide range of chemical functionality useful for conjugation applications.
Polyacetals can also be prepared without generation of a small molecule byproduct that requires removal by the reaction of diols and di-vinyl ethers using an acid catalyst, as described by Heller (J. Heller et al., "Preparation of polyacetals by the reaction of divinyl ethers and polyols", J. Polyp. Sci.: Polyfn. Lett. Ed.,18, 293-297, 1980; J.
Heller et al., "Polyacetal hydrogels formed from divinyl ethers and polyols", US Patent No.
4,713,441, 1987). Such polyacetals have uniform structure in that they are strictly alternating polymers of the A-B type. Uniform structure in biomedical polymer development is critical for optimization of the biological profile and to ensure the polymer meet regulatory requirements. The polymerization of diols and di-vinyl ethers occurs without the elimination of a small molecule under mild conditions. This is more efficient than polymerizations where there is a molecule (e.g. water or methanol) which must be removed.
Polyacetals suitable for conjugation can be prepared by utilization of suitably functionalized diols, di-vinyl ethers, and/or hydroxy-vinyl ether monomers. Thus it becomes possible to prepare polyacetals for conjugation that possess conjugation functionality on either the A or B
monomeric unit. Such structural uniformity is advantageous for controlling conjugation of bioactive molecules along the polymer mainchain.
The production of biodegradable polyacetals derived from polysaccharides which chemically has been described in WO 96/32419. This approach does not give polymeric materials displaying structural uniformity and suffers from the aforementioned limitations where chemical modification (i.e. conjugation of a bioactive molecule) often leads to the polysaccharide to become immunogenic or non-degradable. It is not possible to prepare polymeric materials displaying an alternating A-B structure, rather the structure of these polysaccharide derived polyacetals are too diverse to chemically analyze to the degree necessary to fulfill regulatory requirements.
The disclosures of these and other documents referred to throughout this application are incorporated herein by reference in their entirety.
SUMMARY OF THE INVENTION
The first aspect of the present invention relates to a class of new degradable polymers represented by Formula (I):
R R' ,Y
C~ IC~'O n wherein R and Rl are independently selected from the group consisting of hydrogen, Cl_ls alkyl, Cz_is alkenyl, Cz_ls alkynyl, C6_l8 aryl, C~-18 alkaryl and C~_ls aralkyl groups;
X is a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1_zoo a~~ediyl, C2_zoo alkenediyl, Cz-zoo a~3'nediyl, C6_zoo cycloalkanediyl, C6_zoo cycloalkenediyl, C6_200 _g_ cycloalkynediyl, Cg_ZOO arylenediyl, C6_2oo alkarylenediyl, C6_2oo aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2-10,000.
Another aspect of the invention relates to a bioactive agent, preferably a drug, conjugated to a degradable polymer of Formula (I).
Another aspect of the invention relates to the preparation of the new degradable polymers and new polymer therapeutics.
A further aspect of the invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a polymer-drug conjugate of the present invention in combination with one or more pharmaceutically acceptable carriers.
A further aspect of the invention relates to the use of a polymer-drug conjugate of the present invention in the preparation of a medicament for the treatment of a disease such as cancer.
A further aspect of the invention relates to a method of treatment for diseases such as cancer, comprising administering to a patient in need of such treatment a therapeutically effective amount of a polymer therapeutic of the present invention.
Yet a further aspect of the present invention relates to prepolymers useful for the preparation of the degradable polymers.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 is a graph showing the superposition of gel permeation chromatography traces for polyacetal 3 dissolved in solution at pH 7 and at 37 °C over a period of 21 days.
Figure 2 is a graph showing the superposition of gel permeation chromatography traces for polyacetal 3 dissolved in solution at pH 5.5 and at 37 °C
over a period of 21 days.
Figure 3 is a graph showing polyacetal 3 degradation shown as percent molecular weight loss at pH values of 7.4, 6.5 and 5.5 versus time.
_g_ Figure 4 is a graph showing red blood cell lysis assay of polyacetal 3.
Figure 5 is a graph showing the cytotoxicity of polyacetal 3 using the B16F10 cell line.
Figure 6 is a graph showing superimposed gel permeation chromatography (GPC) traces of polyacetal 22 from a phosphate buffer solution at pH 7.4 and a solution where the pH was adjusted to 1-2 by the addition of HCl.
Figure 7 is a graph showing the degradation profile of polyacetal 22 at pH 7.4 and 5.5 displaying the loss in molecular weight (Mw).
Figure 8a is a graph showing red blood cell (RBC) lysis of polyacetal 22 and its degradation products over a 1 hour time period . No RBC lysis was observed in this assay for the polyacetal.
Figure 8b is a graph showing red blood cell lysis of polyacetal 22 and its degradation products over a 5 hour time period. No RBC lysis was observed in this assay for the polyacetal.
Figure 9 is a graph showing cytotoxicity of polyacetal 22 using the B 16F10 cell line.
Polyacetal 22 does not display cytotoxicity in this assay.
Figure 10 is a graph showing the labeling efficiency with lasI labeled Bolton-Hunter reagent of polyacetal 22 to give conjugate polyacetal 23. This Figure shows the crude product.
Figure 11 is a graph showing the labeling efficiency with lasl labeled Bolton-Hunter reagent of polyacetal 22 to give conjugate polyacetal 23. This Figure shows the purified conjugate product.
Figure 12 is a graph showing the body distribution of radiolabeled polyacetal conjugate at 5 min and 1 hour. This shows that the polyacetal conjugate remains in the blood without accumulating in the organs shown.
DETAILED DESCRIPTION OF THE INVENTION
I. Definitions Terms used herein are based upon their recognized meanings and should be clearly understood by those skilled in the art.
The term "alkyl" refers to a straight or branched saturated monovalent hydrocarbon radical having the number of carbon atoms as indicated.
The term "alkenyl" refers to a straight or branched unsaturated monovalent hydrocarbon radical having the number of carbon atoms as indicated and the distinguishing feature of a carbon-carbon double bond.
The term "alkynyl" refers to a straight or branched unsaturated monovalent hydrocarbon radical having the number of carbon atoms as indicated and the distinguishing feature of a carbon-carbon triple bond.
The term "cycloalkyl" refers to a cyclic saturated monovalent hydrocarbon radical having the number of carbon atoms as indicated.
The terms "cycloalkenyl" and "cycloalkynyl" refer to cyclic unsaturated monovalent hydrocarbon radicals. A "cycloalkenyl" is characterized by a carbon-carbon double bond and a "cycloalkynyl" is characterized by a carbon-carbon triple bond.
The term "aryl" refers to a monovalent unsaturated aromatic carbocyclic radical having one or two rings, such as phenyl, naphthyl, indanyl or biphenyl, or to a monovalent unsaturated aromatic heterocyclic radical such as quinolyl, dihydroisoxazolyl, furanyl, imidazolyl, pyridyl, phthalimido, thienyl and the like.
The term "alkaryl" refers to an aryl group substituted with one or more alkyl groups.
The term "aralkyl" refers to an alkyl group substituted with one or more aryl groups.
The term "alkanediyl" refers to a straight or branched saturated divalent hydrocarbon radical having the number of carbon atoms indicated.
The terms "alkenediyl" and "alkynediyl" refer to straight or branched,unsaturated divalent hydrocarbon radicals. An "alkenediyl" is characterized by a carbon-carbon double bond and an "alkynediyl" is characterized by a carbon-carbon triple bond.
The term "cycloalkanediyl" refers to a cyclic saturated divalent hydrocarbon radical having the number of carbon atoms indicated.
The terms "cycloalkenediyl" and "cycloalkynediyl" refer to cyclic unsaturated divalent hydrocarbon radicals. A "cycloallcenediyl" is characterized by a carbon-carbon double bond and a "cycloalkynediyl" is characterized by a carbon-carbon triple bond.
The term "arylenediyl" refers to a divalent unsaturated aromatic carbocyclic radical having one or two rings. The term "alkarylenediyl" refers to an arylenediyl substituted with one or more alkyl groups and the term "aralkylenediyl" refers to an alkylenediyl" substituted with one or more aryl groups.
The term "peptide bond" is used in its common accepted meaning.
The term "hydrolytically-labile bond" refers to a bond that is capable of undergoing hydrolysis, such as an ester, amide, acetal, or hydrazone bond. Preferably, the hydrolytically-labile bond is labile under acid conditions.
The term "halo" refers to chloro, bromo, iodo and fluoro atoms.
The term "saccharide" is used in its common accepted meaning. The terms "polysaccharide" and "oligosaccharide" refer to carbohydrate molecules containing more than one saccharide unit.
The term "activating/protecting group" refers to a group in a multifunctional compound which may temporarily activate or temporarily block a reactive site wherein a chemical reaction is to be carried out selectively at a reactive site. The reactive site may be other than the site occupied by the "activating/protecting group." The activating/protecting groups referred to, in the context of the present invention, are those commonly known activating/protecting groups including, but not limited to, activating groups such as N-succinimidyl, pentachlorophenyl, pentafluorophenyl, para-nitrophenyl, dinitrophenyl, N-phthalimido, N-norbornyl, cyanomethyl, pyridyl, trichlorotriazine, 5-chloroquinilino, and protecting groups such as N-(9-fluorenylmethoxycarbonyl) (Fmoc), carbobenzyloxy (Cbz), 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde) and imidazolyl.
The term "prepolymer" refers to a reactant used to make a polymer, that is, to monomers and other subunits from which polymers may be formed.
The term "therapeutically effective amount" refers to the amount which, when administered to a patient for treating a disease, is sufficient to effect such treatment for the disease.
The term "treating" or "treatment" is intended to include inhibiting the disease (i.e., arresting its development) and relieving the disease (i.e., causing regression of the disease).
I. The Degradable Polymers The novel degradable polymers are represented by Formula (I) R R~
~Xw ~ iY CI) ~~O O ~~O n wherein R and R1 are selected from the group consisting of hydrogen, C1_lg alkyl, Cz_ is alkenyl, Cz_l8 alkynyl, C6_i$ aryl, C~-18 alkaryl and C~_1$ aralkyl groups;
X is a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1_zoo a~anediyl, Cz_zoo alkenediyl, Cz-zoo alkynediyl, C6_zoo cycloallcanediyl, C6_zoo cycloalkenediyl, C6_zoo cycloalkynediyl, C6_zoo arylenediyl, C6_zoo alkarylenediyl, C6_zoo aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2-10,000.
I. Presently Preferred Embodiments In a preferred embodiment, R and Rl in Formula (I) are the same and are selected from hydrogen, Cl_lz alkyl, Cz_lz alkenyl, Cz_lz alkynyl, C6_iz aryl, C~_lz alkaryl and C7_iz aralkyl groups; more preferably hydrogen, CI_4 alkyl, Cz_4 alkenyl, Cz_4 alkynyl, C6_lo aryl, C~_ to alkaryl and C~_lo aralkyl groups; most preferably hydrogen, Cl_4 alkyl, and Cz_4 alkenyl.
In a preferred embodiment X is a Cl_z4 alkanediyl group, where the alkanediyl group is optionally substituted within the carbon backbone with one or more groups selected from C(O), C(O)NRI, C(O)O, >NRz', wherein N is bound to two carbon atoms within the carbon backbone and Rz' is hydrogen, or is a group capable of displacement so that N
is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group may comprise a pendant group or groups selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups.
In an embodiment of the invention, the alkanediyl group or the pendant group or groups comprises a primary, secondary, or tertiary amine group.
Most preferably, X does not comprise a saccharide, oligosaccharide or polysaccharide.
In the definition of X, any alkyl group or moiety is preferably C1_1$ alkyl, any alkenyl group or moiety is preferably Cz_l8 alkenyl, any alkynyl group or moiety is preferably Cz_lz alkynyl, any aryl group or moiety is preferably C6_z4 aryl, any alkaryl group or moiety is preferably C~_z4 alkaryl and any aralkyl group or moiety is preferably C~_z4 arallcyl, any cycloalkyl group or moiety is preferably C4_za cycloalkyl, any cycloalkenyl group or moiety is preferably CS_z4 cycloalkenyl, and any cycloalkynyl group or moiety is preferably CS_z4 cycloalkynyl.
Most preferably X is selected from the groups (1V)-(VIII) below:
a a 3 R2w ~ N R3 ~I~ Rz/ \~~~~0/ R M
O R~z ~O/ O Rs a a R ~N ~O N ~R3 N9 Rz/N~lt~~~N~R3 Nil) H ''2 ~H O Rs H
R m O Rs O
R \O a'I~"~~m Ra (VIII) O O
wherein R2 and R3 are independently selected from covalent bonds or C1_l8 alkanediyl groups terminating in OH or vinyl ether;
each R12 is independently selected from synthetic or natural amino acid side chains;
each R4 is independently selected from the group consisting of hydrogen, activating/protecting groups and the groups (IX), (X), (XI) and (XII) O R'3 ~(~
~NH ~ ~~NH
~ ~Im m O R~s R~2 H O R~
~~NH ~ ~~NH PaO
m0 R~s m ~~ ~H
R~2 RS and R6 are selected from the group consisting of -NH2~ -NHR13, -OR13, wherein each R13 is independently selected from hydrogen, Cl_4 alkyl, and activating/protecting groups; and m is an integer of 0-20.
Y preferably is represented by the formula -(C"Ha"O)qC"H2", wherein n is an integer of 2-10, preferably 2 or 3 and q is an integer of 1 to 200.
The molecular weight of the polymer of Formula (I) is preferably in the range of 10,000-100,000.
II. Preparation of the Polymer of Formula (I) The polymer of Formula I
R R' 0~ ~Xw ~ iY (I) ~~O O ~~O n wherein R and Rl are selected from the group consisting of hydrogen, Cl_l8 alkyl, Cz_ 1$ alkenyl, C2_1$ alkynyl, C6_l8 aryl, C~_1$ alkaryl and C~_ls aralkyl groups;
X is a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of Cl_zoo alkanediyl, Cz_zoo alkenediyl, Cz-zoo a~ynediyl, C6_zoo cycloalkanediyl, C6_2oo cycloallcenediyl, Cs-zoo cycloalkynediyl, C6_zoo ~ylenediyl, C6_zoo alkarylenediyl, C6_zoo ~'a~Ylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2-10,000 may be prepared by the reaction of a diol of Formula (II) HO~Y~OH (II) with a divinyl ether of Formula (III) R R~ (III) ~O~ X ~p~
wherein R, Rl, X and Y are as defined above.
The diol of Formula (II) preferably is a polyethylene glycol or polypropylene glycol compound having a molecular weight in the range 100-20,000, more preferably polyethylene glycol having a molecular weight in the range 200-10,000, most preferably polyethylene glycol having a molecular weight in the range 200-5,000, in particular a molecular weight of approximately 200-4,000. Such materials are widely available from such commercial sources as Sigma-Aldrich Corporation (St. Louis, MO) and Shearwater Polymers. Inc.
(Huntsville, AL). It will be understood by one of ordinary skill in the art that the reactant of Formula (II) may also comprise any diol of Formula (II), such as other glycols and diols suitable for use in biomaterials.
The divinyl ether of Formula (III) may be obtained commercially or may be made by any suitable means known in the art. For example, commercially-obtained amino vinyl ether may be combined with methyl esters to provide the divinyl ethers of Formula (III). Similarly, the hydroxy vinyl ether compound is commercially available, and may be used to make polyacetal polymers with ester moieties in the main chain. The methyl esters may comprise, for example, esters such as malonates, imines such as iminodiacetates, and other compounds known in the art. Symmetric, achiral methyl esters are preferred synthetic precursors.
The polymerization reaction may be carried out in a solventless system, although preferably the reaction takes place in the presence of an organic solvent selected from aliphatic or aromatic hydrocarbons, which may be optionally halogenated, ethers (including cyclic ethers), dialkylsulfoxides and alcohols (preferably sterically hindered alcohols, for example secondary or tertiary alcohols). Preferred solvents include tetrahydrofuran (THF), dichloromethane, and toluene. A particularly preferred solvent is toluene.
The polymerization is generally carried out in the presence of a suitable catalyst such as a catalyst for acid-catalysis, for example, hydrochloric acid, sulfuric acid, phosphoric acid, p-toluenesulfonic acid, methanesulfonic acid, acetic acid, n-butyric acid, trifluoroacetic acid or oxalic acid. A preferred catalyst is p-toluene sulfonic acid (p-TSA).
The polymerization is conducted at a temperature of -10 °C-200 °C, preferably 20 °C -120 °C, most preferably between about 25 °C and 60 °C.
I. Polyacetal Conjugates of Bioactive Agents and Their Preparation The functionalized polymers of the invention comprise polymers with bioactive functionality, and thus comprise polymers that include bioactive agents. The degradable polyacetal polymers of the invention comprising bioactive functionality may be formed from substrates that include bioactive agents, from polymers to which bioactive agents are conjugated, and from substrates that combine to form bioactive agents.
A bioactive agent may be attached to X in Formula (I). In a particularly preferred embodiment, when X is (IX), (X), (XI), (XII), or >NR2~, wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, the N atoms of the groups >NR2~, (IX), (X), (XI) and (XII) are, or are capable of being covalently attached to the bioactive agent.
The bioactive agent preferably is a pharmaceutically active agent (a "drug").
Suitable drugs include any drugs for which prolonged action and/or a targeted intracellular delivery is desirable, and include anticancer agents, for example doxorubicin, daunomycin, paclitaxel, taxotere, and the like, most preferably, doxorubicin. Other bioactive agents include polypeptides and proteins. The method of attachment may vary somewhat according to the bioactive agent, as is described below; and a person of ordinary skill in the art will be able, having selected a desired bioactive agent, using their knowledge and the disclosure of this application, to attach the bioactive agent to a degradable polyacetal polymer of this invention, thereby forming a conjugated polymer of this invention.
The attachment of the bioactive agent to the polymer of Formula (I) may be effected by the reaction of the polymer with the bioactive agent or a bioactive agent precursor.
Bioactive agents may be attached to the polymer in any suitable manner.
Preferably the attachment is effected subsequent to the polymerization reaction to produce the degradable polymer of Formula (1).
Attachment of bioactive agents may be effected in other ways as well. For example, attachment may be by linkages comprised of groups that covalently couple and cross-link the agents to the polymers. Such linkages may comprise disulfide linkages or ester bonds, or may be acid-labile linkages such as hydrazone linkage as described by Greenfield et al., Gazzcez"
Res., 50, 6600-6607 (1990), and references therein. Alternatively, the attachment may be via a "protected" disulfide bond that sterically inhibits attack from thiolate ions, such as are found in the coupling agents S-4-succinimidyloxycarbonyl-a-methyl benzyl thiosulfate (SMBT) S and 4-succinimidyloxycarbonyl-.alpha.-methyl-a-(2-pyridyldithio) toluene (SMPT), in the manner disclosed in U.S. Patent No. 6,048,736 to Kosak.
Where the bioactive agent has an amino group, it may be useful to form a reactive carbonate half ester in the polymer, P, P-O-CO-X, wherein X is a good leaving group, using reagents such as carbonyl diimidazole, p-nitrophenyl chloroformate or bis-N-succinimidyl carbonate. The activated polymer P, P-O-CO-X, may then be reacted with the bioactive agent under conditions which do not destroy its activity, leading predominantly to urethane linkages attached through the amino group. For example, carbonyl diimida.zole, can be reacted with terminal hydroxyl groups of the polymer. The reaction mixture may be quenched in aqueous solution at neutral pH and the activated polymer isolated, for example by dialysis or size exclusion chromatography, as disclosed in U.S. Patent No. 5, 468,478 to Saifer et al.
Attachment (conjugation) of bioactive agents may be effected by reaction with polymers or monomers with electrophilic functionality. For example, prepolyrners comprising electrophilic pendant chain functionalized monomers which comprise, for example, diols or bis-vinyl ethers, may be used for this purpose.
The following reaction scheme illustrates one route to the production of a polymer-doxorubicin drug conjugate:
~O~O O~O
O NH
O HO-PEG-OH
o,~
~J0 Scheme 2 In this scheme, PEG is the residue of a polyethylene glycol (without the terminal hydroxy groups), the terminal OH groups of the polyethylene glycol being explicitly shown when the entire glycol is meant.
Bioactive agents that may be attached (conjugated) to the polyacetal polymers of the invention include polypeptides and proteins. Such conjugation may be effected at pendant chains and at terminal groups. For example, conjugated polyacetal polymers of the invention include proteins conjugated with a degradable polyacetal polymer.
I. Administration and Pharmaceutical Composition Compositions comprising the degradable polyacetal polymers, with or without attached bioactive agents, are water-soluble or colloidal suspension compositions suitable for incorporation into pharmaceutical solutions or pharmaceutical compositions, or for delivery to an animal or patient for treatment. For example, polyacetal polymers of the invention are Ho.,.. I , HO O OH O
soluble in water and water solutions, such as saline, phosphate buffered saline (PBS), and other buffered solutions. The polyacetal polymers are soluble in solutions of widely varying pH.
In general, degradable polyacetal polymer compositions will be administered in therapeutically effective amounts by any of the usual modes known in the art.
Degradable polyacetal polymers with attached bioactive agents may be directly delivered to solutions bathing cells, tissues or organs in vitro. Pharmaceutical compositions comprising bioactive agents attached to degradable polyacetal polymers may be administered to an animal, including a human, by one of the following routes: oral, topical, systemic (e.g. transdennal, intranasal, or by suppository), or parenteral (e.g. intramuscular, subcutaneous, or intravenous injection). Compositions may take the form of tablets, pills, capsules, semisolids, powders, sustained release formulations, solutions, suspensions, elixirs, aerosols, or any other appropriate compositions; and comprise a bioactive agent attached to a degradable polyacetal polymer of the invention in combination with at least one pharmaceutically acceptable excipient. Suitable excipients are well known to persons of ordinary skill in the art, and they, and the methods of formulating the compositions, may be found in such standard references as Alfonso AR: Remivcgto~'s Pharmaceutical Sciehees, I7th ed., Mack Publishing Company, Easton PA, 1985. Suitable liquid carriers, especially for injectable solutions, include water, aqueous saline solution, aqueous dextrose solution, and glycols.
Pharmaceutical formulations comprising bioactive agents attached to degradable polyacetal polymers of the invention can be prepared according to any method known to the art for the manufacture of pharmaceuticals. Such formulations can contain sweetening agents, flavoring agents, coloring agents and preserving agents. Any such formulation can be admixed with nontoxic pharmaceutically acceptable excipients which are suitable for manufacture.
Pharmaceutical formulations for oral administration can be formulated using pharmaceutically acceptable carriers well known in the art in dosages suitable for oral administration. Such carriers enable the pharmaceutical formulations to be formulated in unit dosage forms as tablets, pills, powder, dragees, capsules, liquids, lozenges, gels, syrups, slurries, suspensions, etc. suitable for ingestion by the patient.
Intravenous injectable compositions are comprised of a polymer-drug conjugate of the invention in combination with at least one pharmaceutically acceptable liquid carrier.
Acceptable liquid carriers are non-toxic, aid administration, and do not adversely affect the therapeutic benefit of the polymer-drug conjugate. Such suitable carriers include, but axe not limited to, water, saline, aqueous dextrose and glycols. Further, excipients and other agents may be included in pharmaceutical compositions along with the degradable polyacetal polymers and attached bioactive agents. In addition, other additives and agents, such as antioxidants, antiseptic or antibiotic agents, buffers, stabilizers, solubilizers and other agents, may be added to degradable polyacetal polymer compositions of the invention.
For example, dimethylsulfoxide, benzoic acid, ascorbic acid, or tocopherol may be included in pharmaceutical compositions comprising degradable polyacetal polymer compositions of the invention. Thus, injectable compositions comprising bioactive agents conjugated to degradable polyacetal polymers will preferably comprise water or saline solutions or emulsions, pharmaceutically acceptable Garners, and may further comprise buffering agents, such as phosphate buffer or HEPES buffer, and optionally other agents.
In general, the polymer-drug conjugates of the invention will be administered in therapeutically effective amounts via intravenous injection. A therapeutically effective amount may vary depending on the severity of the disease, the age and relative health of the subject, the potency of the conjugate used and other factors. A
therapeutically effective amount may range from about 0.001 milligram per Kg (mg/Kg) body weight per day to 100 mg/Kg body weight per day. Preferably the amount will be about 0.1 to 10 mg/Kg/day.
Therefore, a therapeutically effective amount for a 70 Kg patient may range from about 0.07 to 7000 mg/day, preferably about 7 to 700 mg/day. A person of ordinary skill in the art of treating diseases such as cancer will, without undue experimentation, having regard to that skill and this disclosure, be able to determine a therapeutically effective amount of a particular bioactive agent attached to a degradable polyacetal polymer for practice of tlus invention.
The degradable polyacetal polymers of the invention, with or without attached bioactive agents, may be dried or lyophilized and stored in that condition for a considerable length of time without significant degradation or decomposition. Such dried or lyophilized compositions may be reconstituted for use, e.g., for injection, at a convenient time after storage by addition of an appropriate amount of a suitable liquid, preferably a buffered water solution, such as saline. An appropriate amount is that amount sufficient to provide the desired volume so as to result in a solution of the desired final concentration.
Excipients useful for preparation of lyophilized or freeze-dried compositions include saccharides, amino acids, and salts such as inorganic salts. Saccharides may be, for example, monosaccharides, such as glucose and fructose, disaccharides such as maltose, lactose, and sucrose, polysaccharides such as dextran and starch, and sugax alcohols, such as mannitol sorbitol and glycerol. Amino acids may include, for example, glycine, and salts may include, for example, sodium chloride and potassium chloride. Such excipients may be used alone or in combination, and may be useful for inhibiting aggregation in the reconstituted polymer solution.
The amount of a polymer-drug conjugate of the invention in the composition may vary. In general, the final composition will comprise from about 0.001% w/w to 30 % w/w of the polymer-drug conjugate, preferably about 0.01 % w/w to 10% w/w, more preferably about 0.1 % w/w to 5 % w/w with the remainder being the caxrier or carriers.
I. Pharmacology and Utility Degradable polymers are useful in a wide variety of pharmaceutical and biomedical applications. Uses for degradable polymers include coatings for drug tablets, contact lens coatings, coatings for surgical implants and medical devices, gels, as ingredients in topical and optical pharmaceutical solutions, in pharmaceutical formulations including delayed release pharmaceutical formulations and in targeted drug formulations.
Conjugation of bioactive agents, such as anticancer drugs, with degradable polymers helps to enhance the efficacy of the bioactive agent.
Degradable polyacetal polymers of the invention are suitable for use in pharmaceutical and biomedical applications with superior properties compared to prior materials. The polyacetal polymers of the invention are degradable under physiological conditions on a time-scale suitable fox effective delivery of bioactive agents in an animal. In addition, the biodistribution of degradable polyacetal polymers of the invention within the body and bloodstream of the animal receiving the polymer is favorable for the effective delivery of bioactive agents for the treatment of many diseases. The polymers and bioactive agents remain in the bloodstream for hours, not minutes, do not preferentially go to the liver, but remain in circulation so as to provide for the prolonged action of the bioactive agents, and are not toxic.
The stability of the degradable polyacetals of the invention differs in solutions of different pH. Degradable polyacetals of the invention are quite stable in water solutions near neutral pH, less so in more acidic solutions. As shown in Figure 1, polyacetal 3 (at pH 7 and at 37 °C) was quite stable, with little change in the molecular weight at time points ranging from 1 to 505 h (21 days). However, polyacetal 3 was less stable when dissolved at pH 5.5 at 37 °C. As shown in Figure 2, polyacetal 3 had essentially degraded to the PEG monomeric units by 155 h (6.5 days). The results of three such stability experiments are shown in Figure 3, in which the degradation of polyacetal 3 over time is shown as the percent molecular weight loss (Mw) at each pH value vs. time for the pH 5.5, 6.5 and 7.4.
The polyacetal polymers of the invention axe not toxic to cells. This is shown in Figure 4, which present the results of a red blood cell (RBC) lysis assay of amino polyacetal 3. No RBC lysis was observed in this assay for polyacetal 3. Dextran was the control which did not display RBC lysis, while polyethylene imine) (PEI) was the control which caused RBC lysis. In addition, direct measurements of cell toxicity on cells in culture resulted in no measured cytotoxicity. As shown in Figure 5, the cytotoxicity of polyacetal 3 was measured using the B16F10 cell line. Polyacetal 3 did not display cytoxicity in this assay compared to polylysine which was used as a cytotoxic control. Dextran was used as a noncytotoxic control.
Similar to the results shown with polyacetal polymer 3 polyacetal polymer 22 is also sensitive to pH. This is shown in Figure 6, in which are shown superimposed GPC traces of amino polyacetal 22 from a phosphate buffered solution at pH 7.4 and a solution where the pH was adjusted to pH 1-2 by addition of HCI. The polyacetal 22 completely degraded within minutes upon exposure at pH 1-2 to give the trace shown by the arrow labeled pH 1-2. This GPC trace is consistent with the molecular weight of PEG3,4oo. The degradation profile of amino polyacetal 22 at pH 7.5 and pH 5.5 is shown in Figure 7, where the loss in molecular weight is shown as a function of time. The degradation study shown in Figure 7 was conducted at 37 °C and at a concentration of 3 mg/ml of polyacetal 22;
three separate samples were analyzed at each pH. Polyacetal 22 degrades more rapidly in the mildly acidic medium at pH 5.5 than in the relatively neutral physiological pH 7.4. Lysosomal pH is in the range of pH 5.0 to pH 5.5. The experimental pH value of 5.5 was selected to match the Iysosomal pH, which would be that encountered by a physiologically soluble polymer conjugate upon cellular uptake by endocytosis within the lysosome.
The in vitro biocompatibility of amino polyacetal 22 is shown in Figures 8a, 8b, and 9. Figure 8a shows the results of red blood cell lysis experiments at 1 hour, and Figure 8b shows the results of red blood cell lysis experiments at 5 hours. Figure 9 shows the results of cytotoxicity experiments. These experiments show that polyacetal 22 is not lytic or toxic to these cells, and so has a favorable biocompatibility profile.
Polyacetal polymers of the invention do not cause lysis of red blood cells.
The results of a red blood cell (RBC) lysis assay of amino polyacetal 22 and its degradation products is shown over a 1 hour time period in Figure 8a. The degradation products were obtained by dissolving polyacetal 22 in phosphate buffered saline (PBS), adjusting the pH
to 1-2 by the addition of HCl to allow the polyacetal to degrade, then adding a small amount of NaOH to readjust the pH to 7.4. No RBC lysis was observed in this assay for polyacetal 22. Dextran was used as a control; it did not display RBC lysis. Polyethylene imine) (PEI) was also used as a control; it caused RBC lysis. Similarly, in Figure 8b, the results of a red blood cell lysis assay of amino polyacetal 22 and its degradation products are shown over a 5 hour time period. No RBC lysis was observed in this assay for polyacetal 22. Dextran and polyethylene imine) (PEI) were used as control compounds again: dextran did not display RBC
lysis, while PEI did cause RBC lysis. Thus, the results shown in Figures 8a and 8b demonstrate that the polyacetal polymer of the invention, polyacetal 22, was not lytic for red blood cells for up to five hours.
In addition, polyacetal polymers of the invention are not cytotoxic. A
cytotoxicity assay of amine pendant chain polyacetal 22 using B16F10 cell Iine is shown in Figure 9. As shown in Figure 9, polylysine is cytotoxic at concentrations well below 0.1 mg/ml. However, polyacetal 22 did not display cytoxicity in this assay at concentrations of up to several mg/ml, and thus is extremely well tolerated by these cells as compared to the cytotoxic control compound, polylysine. Dextran, used as a noncytotoxic control, was also found not to be cytotoxic even at relatively high concentrations.
Polyacetals of the invention remain in circulation in the blood with relatively little loss from the blood circulation to the organs. lzsl labeled polyacetal polymers of the invention may be formed using Bolton-Hunter methods, as shown in Figures 10 and 11 and described in Example 7. In the body distribution study shown in Figure 12, polyacetal 23 was predominantly in the blood, at both at 5 min and at 1 hour after administration. The continued presence of polyacetal 23 in the blood in significant amounts at one hour is a surprising and favorable property of the polyacetal of the invention. This study shows that polyacetal 23 remains in the blood without accumulating in the organs shown. In particular, and in contrast to many previously-known polymers, the polyacetal of the invention is not significantly taken up by the liver but remains substantially in circulation in the blood for an hour. Thus, the polyacetal of the invention possesses the favorable properties of long-duration presence in the blood, of very little removal from circulation by the organs, and, since very little of the polyacetal is lost to the liver, relatively little degradation by the liver.
Functionalized polymers of the invention, such as may be formed by synthesis from fwctionalized precursors or by attaclnnent of bioactiee agents, such as anticancer drugs, to degradable polyacetal polymers of the invention may be effective to enhance the efficacy of the bioactive agent. As shown in Figure 12, polyacetal polymers remain in circulation with relatively little removal or loss from the blood on a timescale of hours, a property which enhances the effectiveness of anticancer drugs attached to degradable polyacetal polymers of the invention. Higher dosages of the anticancer drugs, which are more effective in treating the cancerous tissue than lower dosages, are tolerated by animals when anticancer drugs are attached to degradable polyacetal polymers of the invention.
I. Prepolymers of Formula (XIII) The prepolymers are novel divinylethers represented by Formula (XIII) ' (X111) wherein R' and R8 are selected from the same groups as R and Rl, Z is a C1_z4 alkanediyl group, optionally substituted within the carbon backbone with one or more or a mixture of the groups selected from carbonyl, peptide, ester, >NRz', wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group comprises a pendant group R9, wherein R9, is selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloallcenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups.
In a preferred embodiment, Rg contains at least one activating/protecting group.
In the definition of Z, any alkyl group or moiety is preferably Cl_l8 alkyl, any alkenyl group or moiety is preferably C2_lg alkenyl, any alkynyl group or moiety is preferably C2_lz alkynyl, any aryl group or moiety is preferably C6_24 aryl, any alkaryl group or moiety is preferably C~_Z4 alkaryl and any aralkyl group or moiety is preferably C~_24 aralkyl, any cycloalkyl group or moiety is preferably C4_z4 cycloalkyl, any cycloalkenyl group or moiety is preferably CS_24 cycloalkenyl, and any cycloalkynyl group or moiety is preferably CS_24 cycloalkynyl. Symmetric achiral methyl esters are preferred synthetic precursors.
Preferably Z has a structure (X1V), (XV), (XVI), (XVII) and (XVIII), below:
OI1 R9 O L. O
R' ~O~j~ ~Rm (XIV) Rz/0~~~~~O~Rm lRY~z ~O IO Y R~ a O R9 ' O Ii O
R~ N /R~~ ~1) RzrN~\~~~HrRt~ ~If) N~ ~ Ip1'1 ~,R~ '9 O Rs O
R~ ~O'~u~~0 O m X111) P O P P P R
O
wherein Rl° and Rll are selected from covalent bonds, and Cl_6 allcanediyl groups;
R9 is as defined above, R12 are selected from synthetic or natural amino acid side chains; and p is an integer of 0-20.
Prepolymers as disclosed herein may be formed by any suitable method, including such methods as are disclosed in the Examples, such as Examples 3, 4 and 5 following.
A process for the preparation of a prepolyrner of Formula (XII~ may comprise the following steps. To prepare a prepolymer of Formula (XIII):
R' R9 ~ (X111) ~Oit~O
wherein R~ and R8 are selected from the group consisting of hydrogen, C1_l8 alkyl, CZ_la alkenyl, CZ_l8 alkynyl, C6_l8 a aryl, C~_l8 alkaryl and C~_l8 aralkyl groups;
Z is a Cl_z4 alkanediyl group, where the alkanediyl group is optionally substituted within the carbon backbone with one or more groups selected from C(O), C(O)NRI, C(O)O, >NRZ~, wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group comprises a pendant group R9 selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoall~yl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups;
the steps of the process comprise:
reacting a methyl ester of Formula (XIX) O (XIX) H3C0 X' \OCH3 with a vinylether of Formula (XX) H2N ~YwO~ (~) or Formula (XXI) H O~YwO~ (XXI ) wherein X comprises a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond, and Y is a group selected from the group consisting of linear and branched C1_zoo alkanediyl, Cz_zoo alkenediyl, Cz_zoo a~ynediyl, C6_zoo cycloalkanediyl, C6_zoo cycloallcenediyl, C6_zoo cycloalkynediyl, C6_zoo az'Ylenediyl, C~_zoo alkarylenediyl, C~_zoo. aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone.
In a preferred embodiment of the method of preparing a prepolyrner of Formula (XIII) R9 contains at least one activating/protecting group.
These prepolymers are particularly useful for the preparation of the polymers of Formula (I) by methods known in the art and as illustrated in the Examples.
Functionalized prepolymers, such as prepolymers functionalized with bioactive agents or precursors to bioactive agents, are useful for the preparation of polymers of formula comprising bioactive agents. The following structures represent some particularly preferred prepolymers of the present invention.
O
N O O~OW% ~'O
O O
~O
~~''O
O O HN O
'~ ~,O O p O
O ~ ~ ~ U
O O
~O~N ~~0~
H
NHFmoc O O
~O~~N N~O~
H H
NH
O
Cbb-I ),N
NHFmoc O
O ~ O
~O~N~N~N~O~
H , H
Scheme 1 EXAMPLES
General The degradable polymers of the present invention may be prepared by the reaction of polyethylene glycol) (PEG) as the source of diol (PEG' S with molecular weights of 3,400 g/mol were used) and commercially available triethylene glycol di-vinyl ether.
PEG is selected as the diol because it is generally recognized as safe (GR.AS) by drug regulatory authorities and is widely used in pharmaceutical formulation. However, it will be appreciated by those of ordinary skill in the art that other diols, including PEGs of lower or higher molecular weight, are also suitable for the practice of the invention. The use of the tmfunctionalized divinyl ether, triethylene glycol di-vinyl ether, in the preliminary experiments was conducted to confirm a suitable degradation profile (needed for lysosomal degradation) and to confirm ih vitro biocompatibility. Tt will be understood by one of ordinary skill in the art that degradable polyacetal polymers of the invention may also be prepared from functionalized starting materials. For example, functionalized vinyl ethers, particularly functionalized divinyl ethers, may be used as starting materials in the preparation of the degradable polyacetal polymers of the invention. Each following experimental example is preceded by a scheme summarizing the reaction involved. In each case m is an integer representing a PEG molecule of the identified molecular weight Mn.
Example 1. Synthesis of polyacetal 3 by polymerization in toluene HO~O~H + ~O~O~O
ll '''' J m _1 2 ~, pTSA (cat)/toluene O O~ O~O~O~O~O
-m1i n Polyethylene glycol) (Mn = 3,400 g/mol, 17.0 g, 5.0 mmol, 1.0 ec~, para-toluenesulfonic acid monohydrate (0.03 g, 0. 15 mmol, 0.03 ec~ and toluene (60 ml) were added to a 100 ml round bottom flask which was equipped with a stirring bar and fitted with a thermometer, Dean Starlc trap and condenser. An azeotropic distillation of the stirred toluene solution (oil bath, T=150 °C) under nitrogen proceeded for two hours.
The solution was then allowed to cool to ~ 50 °C and tri(ethylene glycol) divinyl ether (1.073 g, 1.083 ml, 5.2 mmol, 1.04 eq) was added by syringe. Within one minute the reaction mixture became visibly more viscous and after 15 minutes the viscosity appeared to be very high.
Toluene (30.0 ml) was added to decrease the viscosity and the clear colorless reaction mixture was stirred a further 2 hours at ambient temperature. Aqueous NaHC03 (8.0%, 2.0 ml) was added to the reaction mixture which was then rapidly stirred for 15 minutes. The aqueous phase was allowed to settle and the toluene phase was carefully decanted into stirred hexane (200 ml) to precipitate the polyacetal. After stirring in the hexane for an additional 10 minutes the polyacetal was collected and placed into a fresh solution of hexane and stirred for a further 10 minutes. The polyacetal was again collected and then dried in vacuum at 50 °C for 4 hours to give a white fluffy solid. The molecular weight was determined to be Mw=42,806 g/mol, Mn=26760 g/mol; polydispersity-1.60 by GPC. The GPC was calibrated with PEG
standards;
56,000, 23,500 and 5598 g/mol.
Example 2. Synthesis of polyacetal 3 by polymerization in THF
A suspension of PEG3aoo (1.041 g, 0.306 mmol) and para-toluenesulfonic acid (25 mg) in toluene (50 ml) was heated to reflux; the flask was fitted with a Dean and Stark trap and a balloon of argon was fitted to the condenser. After 150 min most of the toluene was distilled off. To the residue was added the divinyl ether (0.306 mmol, 1.0 eq) in freshly distilled THF (10 ml). The mixture was stirred at room temperature under argon for 16h.
Triethylamine (0.2 ml) was added and the mixture was stirred for 5 minutes.
The mixture was poured into hexane (300 ml) with rapid stirring, after 5 min the hexane was decanted off and the residue was washed with further hexane (200 ml) for 30 minutes. The polymer was filtered off.
This same procedure was used for polymerizations conducted in dichloromethane.
Exam 1p a 3. Synthesis of bis-vinyl ethers useful for preparing polyacetals The vinyl ethers were made from methyl esters using the commercially available amino vinyl ether. This avoided the use of heavy metals to make the vinyl ether moiety.
~ ~ !
CH30- v 'OCH3 '~ H2N O ~ > // _O NH NH O
s A solution of 3-amino-1-propanol vinyl ether 5 (0.27 mmol, 2.2 eq) and dimethyl malonate 4 (0. 12 mmol, 1 eq) in dichloromethane (S.0 ml) was stirred at ambient temperature for 3 days. The reaction mixture was diluted with dichloromethane (SO ml) and washed with water (2 x 35 ml), conc. NaCI solution (3S ml) and dried over MgS04. The solvent was evaporated to give a semi-solid residue which was triturated with ether-hexane (1:l) to give the bis vinyl ether 6.
Exam 1p a 4. Synthesis of bis-vinyl ethers useful for preparing polyacetals with functionality to conjugate bioactive compounds H3CO~~~OCH3 + ~O~NH, > ~p~NH~~NH~O~
A saturated aqueous Na2C03 solution (20 ml) of dimethyl iminodiacetate 7 (10 mmol, 1.0 eq) and 3-amino-1-propanol vinyl ether 5 (40 mmol, 4.0 eq) was stirred for 1 h at 90 °C.
The solution was cooled and extracted three times with ethyl acetate (SO ml each time). The organic layer was washed with brine, dried over MgS04, and rotoevaporated to give the bis-vinyl ether 8 as a white crystalline solid.
The bis-vinyl ether 8 was then allowed to react with various acylating agents (e.g.
Fmoc protected glycine N-hydroxysuccinimde ester and benzyl chloroformate) to protect (block) the amino functionality in 8 prior to polymerization. The protecting (blocking) group is important because it allows polymerization to proceed without competitive side reactions with the conjugating fimctionality. After polymerization it must be removed without causing degradation of the polyacetal. This strategy is illustrated in Example 5 for the preparation and polymerization of glutamic acid derived bis-vinyl ether 11 and the subsequent deprotection of the polyacetal.
Exam 1p a 5. Synthesis of bis-vinyl ethers useful for preparing polyacetals with a protected primary amine functionality ~~11). Preparation of a amino pendant chain functionalized polyacetal using a pendant chain functionalized bis-vinyl ether monomer (11-X13) Synthesis of Fmoc-glutamyl chloride 10 ~9-X10) \ ~ ~,/~~ ~~2 HO~~~OH > CI~~~~~CI 5 ~O~NH NH~O
INHFmoc INHFmoc NHFmoc _9 _10 _11 To a suspension of Fmoc-glutamic acid 9 (3.13g, 8.5 mmol) in anhydrous CH2C12 (50 ml) was added oxalyl chloride (5.0g, 39 mmol). The mixture was cooled to 0 °C and DMF (2 drops) was added. The mixture was stirred at 0 °C for 1 h then at ambient temperature for 1 h under argon atmosphere. Freshly distilled THF (6.0 rnl) was added and the mixture was stirred at room temperature for 1 h under argon. Hexane (400 ml) was added and the mixture stirred at ambient temperature for 30 minutes. The hexane was decanted off and the residue recrystallized from CHaCl2/hexane to give 10 as white crystals (1.41g).
Synthesis of Fmoc glutamic acid divinyl ether 11 10-X11 To the bis-acid chloride 10 (1.41 g, 3.47 mmol) in anhydrous CH2C12 (25 ml) was added a solution of 3-aminopropyl vinyl ether 5 (701 mg, 6.94 mmol) and NaHC03 in water (10 ml) dropwise over 5 min with vigorous stirring at 0 °C. The mixture was stirred at 0 °C
for 10 min then at ambient temperature for 1 hour. The mixture was diluted with CHaCIa (200 ml) then washed with 2% aqueous NaHC03 (150 ml), brine (150 ml) and then dried with MgS04. Evaporation gave a pale brown solid which was recrystallized from isopropanol/hexane to yield the divinyl ether 11 as an off white solid (910 mg).
Synthesis of amino-functionalized bis-vinyl ether monomer 13 (1113) O O HO~O~H
I. '" J m o 0 G~~r~~r~~c~ 1 _ NHFmoc ~ ~~~NH~NH~
_i l m-1 NHFmoc m-1 NHz A suspension of PEG34oo 1 (1.041 g, 0.306 mmol) and p-toluene sulfonic acid (25 mg) in toluene (50 ml) was heated to reflux in a round bottom flask fitted with a Dean and Stark trap to collect the water. After no further increase in water collection was observed, most of the toluene was distilled from the round bottom flask. To the residue was added the divinyl ether 11 (164 mg, 0.306 mmol) in freshly distilled (sodium-benzophenone) THF
(10 ml). The mixture was stirred at room temperature under argon for 16 h. Triethylamine (0.2 ml) was added and the mixture was stirred for 5 minutes. The mixture was poured into hexane (300 ml) with rapid stirring to precipitate the polyacetal 12. After 5 min the hexane was decanted off and the polyacetal 12 was stirred with fresh hexane (200 ml) for 30 min.
The polyacetal 12 was filtered, collected and dried in vacuum. The weight average molecular weight as determined by GPC (eluent: water, 1 mI/min; PEG standards) was 14300 g/mol.
The Fmoc group was removed by dissolving the polymer into an dichloromethane (7% weight percent) followed by the addition of morpholine and the reaction stirred for 15 min at ambient temperature. The amino functionalized polyacetal 13 was precipitated into a stirred solution of hexane (200 ml).
Example 6. Synthesis of a symmetric, achiral bis-vinyl ether 16 with a protected primary amine useful for preparing polyacetals with functionality to conjugate bioactive compounds ~ ~ ~~NHZ
H3C0' Y'OCHg Cbz-Gly-NHS> H3CO~OCHg 5 > ~O~NH~NH"~~O~
INH3+Cl- H H
O O
14 cb~Hrr 15 cbzHrr 16 A solution of Cbz-gly-NHS (0.75 g, 2.5 mmol), dimethyl aminomalonate hydrochloride 14 (0.45 g, 2.5 mmol) and triethylamine (0.375 ml) in dichloromethane (5 ml) was stirred in a 25 ml single neck round bottomed flask at ambient temperature for 12 hours.
A white precipitate assumed to be triethylamine hydrochloride was evident. The reaction mixture was diluted in dichloromethane (80 ml) and transferred to a separatory funnel, and washed with water and brine. The organic layer was dried over MgS04 filtered and the solvent removed by rotoevaporation to give the amino acid dimethylmalonate 15 as white solid (83%). Synthesis of bis-vinyl ether 16 was completed in dichloromethane by the same process as for the preparation of bis-vinyl ether 8 (Example 4).
Exam 1p a 7. Preparation and in vitro biocompatibility evaluation of a pendant chain functionalized polyacetal 22.
This example describes the preparation of a pendant chain functionalized polyacetal.
This polyacetal 22 was prepared by a terpolymerization process using a suitably functionalized diol monomer 20 for the incorporation of the pendant chain functionality onto the polyacetal. A radiolabel agent was then conjugated to polyacetal 22 to give the labelled conjugate 23 which was used in an ih vivo biodistribution study. An iya vitf o degradation study is also described for polyacetal 22 in this example.
Synthesis of a diol 20 that is functionalized to provide the pendant chain in the final polyacetal HO~ OH ' HO~ OH
HN O
To a rapidly stirred solution of amino diol 19 (1.0 g, 10.0 mmol) and NaOH (1 M, 25 ml) that was cooled to 0-2 °C with a water ice bath was slowly added a dichloromethane (10 ml) solution of Fmoc-chloride (3.4 g, 13.1 mmol, 1.2 eq) over a 1 h period.
The solution was stirred a further 1 h at 0 °C then at ambient temperature for 4 1i. The reaction mixture was transferred to a rotoevaporator and the dichloromethane was evaporated off. To the aqueous residue was added ethyl acetate (70 ml), the solution transferred to a separatory funnel and the organic layer washed with dilute aqueous HCl solution (5 %), dilute NaHC03, brine, dried over MgS04 and rotoevaporated to give a solid which was recrystallized in chloroform to give the Fmoc protected amino diol 20.
Terpolymerization process to prepare the protected amino functionalized polyacetal 21. This is a polymeric precursor to the desired amine functionalized polyacetal 22 H~ OH
HNI O
2 +
_20 H~-PEG-OH
PEG3,4oo 1 (5.005 g, 1.47 mmol, 1.0 eq) andp-toluene sulfonic acid (0.012 g) were added to a 100 ml single neck round bottom flask equipped with a stir bar.
This mixture was heated in vacuum (0.5-1.0 torn) at a temperature of 80-90 °C (oil bath) for 3.0 h. After cooling the flask was purged with nitrogen and Fmoc serinol 20 (0.461 g, I.47 mmol 1.0 eq) and freshly distilled THF (10.0 ml; distilled from sodium-benzophenone) were added to the flask.
To this stirred solution was added a solution of tri(ethylene glycol) 2 (0.601 ml, 2.94 mmol, 2.0 eq) in THF (10.0 ml). The reaction mixture was vigorously stirred at ambient temperature for 2 hour, then triethylamine (0.3 ml) was added to deprotonate the p-toluene sulfonic acid.
The reaction mixture was slowly added to a stirred solution of hexane (I00 ml) to precipitate the polyacetal 21 which was filtered, collected and dried in vacuum at ambient temperature.
GPC analysis indicated the molecular weight was about 25,000 g/mol (eluent:
phosphate buffer solution, 1 ml%min; 2 Waters Hydrogel Columns; PEG standards). H-NMR
analysis confirmed the equivalent incorporation of the Fmoc-amino diol monomer 20 and PEG into the polyacetal 21. The removal of the Fmoc group is given below.
~G~O-PE6-(~~ i eridin ~
[, J n o-PEA
3 n 21 ' 22 A solution of polyacetal 21 (2.050 g) in 20 % piperidine in dichloromethane (10 ml) was stirred at ambient temperature. Thin layer chromatography was used to monitor the reaction (eluent: ethyl acetate). The amino functionalized polyacetal 22 was isolated by first partitioning the piperidine and an unknown amount of by-products into hexane then the dichloromethane was evaporated. The residue was dissolved in THF then this solution added to a stirred solution of hexane (100 ml) to precipitate the desired amino polyacetal ~22. GPC
analysis indicated the polymer molecular weight to be about 23,000 g/mol (eluent: phosphate buffer solution 1 ml/min; 2 Waters Hydrogel Columns; PEG standards). 1H-NMR
analysis indicated the loss of the aromatic blocking group with no reduction in the acetal functionality.
Conjugation of an electrophilic compound to polyacetal 22 (~ ~ '~~ Bolton-Hunter (~ ~ '~~
~~O-PECr-~~ R~ C 'LT( 'O-PEG-J 3 NHz JJ 3 3 _22 _23 OH
The Bolton-Hunter (N-succinimidyl 3-(4-hydroxy 5-[lzsl]iodophenyl) propionate) method may be used to iodinate peptides which do not contain a tyrosine residue or peptides whose activity is destroyed by tyrosine iodination. See, for example, A.E.
Bolton et al., Biochem. J.,133, :529-38, 1973. The conjugation of Bolton-Hunter radiolabel reagent to the amine fiuictionalized polyacetal 22 gives the conjugated polyacetal 23 which is radiolabeled with lzsI.
The amino polyacetal 22 was radiolabeled using the Bolton Hunter reagent by first dissolving polyacetal 22 (50 mg) at 10 mg/mL in borate buffer (0.1 M) at pH
8.5 by the addition of a small amount of NaOH. To this stirred solution was added a solution of the Bolton Hunter reagent (500 ~Ci) in benzene with 2% v/v DMF. The reaction mixture was stirred for 15 minutes at ambient temperature and a small aliquot removed (10 p1) as an archive sample and to determine labeling efficiency. The remaining reaction mixture was diluted with phosphate buffer solution (PBS) to 10 ml, transferred to dialysis tubing (molecular mass cut-off 1000 g/mol) and dialyzed against water (5.01) until no radioactivity was found in the dialysate. The water was changed twice daily over a three day period.
Following dialysis the quantity of free iodide [lzsI~ remaining in the preparation was determined by paper electrophoresis and the 10 ml solution containing the radiolabeled polyacetal 23 was transferred to a vial and stored at -18 °C.
The labeling efficiency as determined by paper electrophoresis is shown in Figures 5 (crude product) and 6 (purified product 23). The body distribution in the rat of the radiolabeled polyacetal 22 is shown in Figure 7.
Following dialysis the quantity of free iodide [lzsI~ remaining in the preparation was determined by paper electrophoresis and the 10 ml solution containing the radiolabeled polyacetal 23 was transferred to a vial and stored at -18 °C.
The labeling efficiency as determined by paper electrophoresis is shown in Figures 5 (crude product) and 6 (purified product 23). The body distribution in the rat of the radiolabeled polyacetal 22 is shown in Figure 7.
Claims (25)
1. A polymer represented by Formula (I), wherein R and R1 are independently selected from the group consisting of hydrogen, C1-18 alkyl, C2-18 alkenyl, C2-18 alkynyl, C6-18 aryl, C7-18 alkaryl and C7-18 aralkyl groups;
X is a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200 aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2 to 10,000.
X is a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200 aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2 to 10,000.
2. The polymer of Claim 1 wherein R and R1 are the same and are selected from hydrogen, C1-4 alkyl, and C2-4 alkenyl.
3. The polymer of Claim 1 wherein X is a C1-24 alkanediyl group, optionally substituted within the carbon backbone with one or more groups selected from C(O), C(O)NR1, C(O)O, >NR2', wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N
is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group may comprise a pendant group or groups selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups.
is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group may comprise a pendant group or groups selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups.
4. The polymer of claim 3, wherein the alkanediyl group or a pendant group comprises a primary, secondary, or tertiary amine group.
5. The polymer of Claim 1 wherein X does not comprise a saccharide, oligosaccharide or polysaccharide.
6. The polymer of Claim 1 wherein X is selected from the groups (IV)-(VIII) wherein R2 and R3 are selected from covalent bonds or C1-18 alkanediyl groups;
R12 are selected from synthetic or natural amino acid side chains;
R4 is selected from the group consisting of hydrogen, activating/protecting groups, and the groups (IX), (X), (XI) and (XII), R5 and R6 are selected from the group consisting of -NH2' -NHR13, -OR14 (wherein R13 and R14 are selected from activating/protecting groups), the groups (IX), (X), (XI) and (XII), and R, wherein R has the same definitions given in Claim 1, and each group may be the same or different; and m is an integer of 0 - 20.
R12 are selected from synthetic or natural amino acid side chains;
R4 is selected from the group consisting of hydrogen, activating/protecting groups, and the groups (IX), (X), (XI) and (XII), R5 and R6 are selected from the group consisting of -NH2' -NHR13, -OR14 (wherein R13 and R14 are selected from activating/protecting groups), the groups (IX), (X), (XI) and (XII), and R, wherein R has the same definitions given in Claim 1, and each group may be the same or different; and m is an integer of 0 - 20.
7. The polymer of Claim 6 wherein X does not comprise a saccharide, oligosaccharide or polysaccharide, and is covalently attached to an activating/protecting group.
8. A polymer-drug conjugate comprising a polymer of Formula (I) wherein R and R1 are independently selected from the group consisting of hydrogen, C1-18 alkyl, C2-18 alkenyl, C2-18 alkynyl, C6-18 aryl, C7-18 alkaryl and C7-18 aralkyl groups;
X is covalently conjugated to a drug via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200~ aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2 - 10,000.
X is covalently conjugated to a drug via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200~ aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2 - 10,000.
9. The polymer-drug conjugate of Claim 8 wherein the drug is an anti-cancer agent selected from a group consisting of doxorubicin, daunomycin, paclitaxel, and taxotere.
10. The polymer-drug conjugate of Claim 9 wherein the drug is doxorubicin.
11. A process for the preparation of a polymer of Formula (I) wherein R and R1 are independently selected from the group consisting of hydrogen, C1-18 alkyl, C2-18 alkenyl, C2-18 alkynyl, C6-18 aryl, C7-18 alkaryl and C7-18 aralkyl groups;
X is a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of linear and branched C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200.
aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2-10,000, which process comprises reacting a diol of Formula (II) with a ivinyl ether of Formula (III), wherein R, R1, X and Y are as defined above.
X is a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of linear and branched C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200.
aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2-10,000, which process comprises reacting a diol of Formula (II) with a ivinyl ether of Formula (III), wherein R, R1, X and Y are as defined above.
12. The process of Claim 11 wherein Y has the formula -(C n H2n O)q,C n H2n-, wherein n is an integer of 2-10 and q is an integer of 1 to 200.
13. The process of Claim 11 wherein the polymerization reaction is carried out in the presence of an organic solvent selected from aliphatic and aromatic hydrocarbons, which may be optionally halogenated, ethers (including cyclic ethers), dialkylsulfoxides, and alcohols.
14. The process of Claim 11 wherein the polymerization is carried out in the presence of a catalyst.
15. The process of Claim 11 wherein the polymerization is conducted at a temperature of about ~10 °C to 200 °C.
16. A process for the preparation of a polymer-drug conjugate comprising a polymer of Formula (I) wherein R and R1 are independently selected from the group consisting of hydrogen, C1-18 alkyl, C2-18 alkenyl, C2-18 alkynyl, C6-18 aryl, C7-18 alkaryl and C7-18 aralkyl groups;
X is covalently conjugated to a drug via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200. aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2 to 10,000, which process comprises reacting a polymer of Formula (I) wherein X is a group capable of being covalently conjugated to a drug, with the drug.
X is covalently conjugated to a drug via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200. aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2 to 10,000, which process comprises reacting a polymer of Formula (I) wherein X is a group capable of being covalently conjugated to a drug, with the drug.
17. The process of Claim 16 wherein the drug is an anti-cancer agent.
18. A pharmaceutical composition comprising a polymer-drug conjugate comprising a polymer of Formula (I) wherein R and R1 are independently selected from the group consisting of hydrogen, C1-18 alkyl, C2-18 alkenyl, C2-18 alkynyl, C6-18 aryl, C7-18 alkaryl and C7-18 aralkyl groups;
X is covalently conjugated to a drug via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200 aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2 - 10,000, in combination with one or more pharmaceutically acceptable carriers.
X is covalently conjugated to a drug via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200 aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2 - 10,000, in combination with one or more pharmaceutically acceptable carriers.
19. The pharmaceutical composition of Claim 18 wherein the drug is an anti-cancer agent.
20. Use of a polymer drug conjugate comprising a polymer of Formula (I) wherein R and R1 are independently selected from the group consisting of hydrogen, C1-18 alkyl, C2-18 alkenyl, C2-18 alkynyl, C6-18 aryl, C7-18 alkaryl and C7-18 aralkyl groups;
X is covalently conjugated to an anti-cancer drug via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200. aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2 - 10,000, in the preparation of a medicament having anti-cancer properties.
X is covalently conjugated to an anti-cancer drug via a peptidic or a hydrolytically-labile bond;
Y is a group selected from the group consisting of C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200. aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone; and n is an integer of 2 - 10,000, in the preparation of a medicament having anti-cancer properties.
21. Use according to Claim 20 wherein the drug is doxorubicin.
22. A prepolymer having the structure (XIII) wherein R7 and R8 are selected from the group consisting of hydrogen, C1-18 alkyl, C2-18 alkenyl, C2-18 alkynyl, C6-18 a aryl, C7-18 alkaryl and C7-18 aralkyl groups;
Z is a C1-24 alkanediyl group, where the alkanediyl group is optionally substituted within the carbon backbone with one or more groups selected from C(O), C(O)NR1, C(O)O, >NR2', wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group comprises a pendant group R9 selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups.
Z is a C1-24 alkanediyl group, where the alkanediyl group is optionally substituted within the carbon backbone with one or more groups selected from C(O), C(O)NR1, C(O)O, >NR2', wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group comprises a pendant group R9 selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups.
23. The prepolymer of claim 22, wherein R9 contains at least one activating/
protecting group.
protecting group.
24. The prepolymer of Claim 22, wherein Z is represented by a structure selected from (XIV), (XV), (XVI), (XVII) and (XVIII), wherein R9 is selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups;
R12 is selected from a group consisting of synthetic or natural amino acid side chains;
R10 and R11 are selected from covalent bonds or C1-6 alkanediyl groups; and p is an integer of 0 - 20.
R12 is selected from a group consisting of synthetic or natural amino acid side chains;
R10 and R11 are selected from covalent bonds or C1-6 alkanediyl groups; and p is an integer of 0 - 20.
25. A process for the preparation of a prepolymer of Formula (XIII) wherein R7 and R8 are selected from the group consisting of hydrogen, C1-18 alkyl, C2-18 alkenyl, C2-18 alkynyl, C6-18 a aryl, C7-18 alkaryl and C7-18 aralkyl groups;
Z is a C1-24 alkanediyl group, where the alkanediyl group is optionally substituted within the carbon backbone with one or more groups selected from C(O), C(O)NR1, C(O)O, >NR2', wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group comprises a pendant group R9 selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups, wherein R9 contains at least one activating/protecting group.
which process comprises reacting a methyl ester of Formula (XIX), with a vinylether of Formula (XX) or a vinylether of Formula (XXI) wherein X comprises a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond, and Y is a group selected from the group consisting of linear and branched C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200.
aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone.
Z is a C1-24 alkanediyl group, where the alkanediyl group is optionally substituted within the carbon backbone with one or more groups selected from C(O), C(O)NR1, C(O)O, >NR2', wherein N is bound to two carbon atoms within the carbon backbone and R2' is hydrogen, or is a group capable of displacement so that N is capable of linking to a bioactive agent, or is a group capable of linking to a bioactive agent, -O- and -S-, and the alkanediyl group comprises a pendant group R9 selected from haloaryl, haloalkyl, aryl, alkaryl, aralkyl, alkoxyaryl, alkoxyalkyl, aminoalkyl, mono-, di- and tri-alkylaminoalkyl, arylaminoalkyl, aminoacyl, N-aryl-N-alkylaminoalkyl aminoaryl, alkoxy, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, cycloalkynyloxy, haloalkoxy, aralkoxy, alkoxyaryloxy, alkoxyalkoxy, aminoalkoxy, mono-, di- and trialkylaminoalkoxy, arylaminoalkoxy, N-aryl-N-alkylamino-alkoxy, acyloxy, acyloxyalkyl, acylaminoalkyl, N-diacyl-iminoalkyl groups, acylamino, alkylamino and hydroxy groups, wherein R9 contains at least one activating/protecting group.
which process comprises reacting a methyl ester of Formula (XIX), with a vinylether of Formula (XX) or a vinylether of Formula (XXI) wherein X comprises a group capable of being covalently conjugated to a bioactive agent via a peptidic or a hydrolytically-labile bond, and Y is a group selected from the group consisting of linear and branched C1-200 alkanediyl, C2-200 alkenediyl, C2-200 alkynediyl, C6-200 cycloalkanediyl, C6-200 cycloalkenediyl, C6-200 cycloalkynediyl, C6-200 arylenediyl, C7-200 alkarylenediyl, C7-200.
aralkylenediyl groups, or any of the above groups wherein the carbon backbone is substituted with one or more oxygen atoms within the carbon backbone.
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Families Citing this family (49)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8106098B2 (en) * | 1999-08-09 | 2012-01-31 | The General Hospital Corporation | Protein conjugates with a water-soluble biocompatible, biodegradable polymer |
AU2001288829A1 (en) * | 2000-09-06 | 2002-03-22 | Ap Pharma, Inc. | Degradable polyacetal polymers |
US6590059B2 (en) | 2001-05-11 | 2003-07-08 | Ap Pharma, Inc. | Bioerodible polyorthoesters from dioxolane-based diketene acetals |
US6939561B2 (en) * | 2001-06-28 | 2005-09-06 | Wisconsin Alumni Research Foundation | Methods and compositions for polyene antibiotics with reduced toxicity |
US7838619B2 (en) * | 2002-01-14 | 2010-11-23 | The General Hospital Corporation | Biodegradable polyketal polymers and methods for their formation and use |
WO2003082303A1 (en) * | 2002-03-29 | 2003-10-09 | Wisconsin Alumni Research Foundation | Polymeric micelle formulations of hydrophobic compounds and methods |
WO2004009774A2 (en) * | 2002-07-19 | 2004-01-29 | Amgen Inc. | Protein conjugates with a water-soluble biocompatible, biogradable polymer |
AU2003254023A1 (en) | 2002-07-19 | 2004-02-09 | The General Hospital Corporation | Oxime conjugates and methods for their formation and use |
US7045589B2 (en) | 2002-11-15 | 2006-05-16 | A.P. Pharma, Inc. | Bioerodible poly(ortho esters) from dioxane-based di(ketene acetals), and block copolymers containing them |
US6878374B2 (en) | 2003-02-25 | 2005-04-12 | Nitto Denko Corporation | Biodegradable polyacetals |
US7332477B2 (en) * | 2003-07-10 | 2008-02-19 | Nitto Denko Corporation | Photocleavable DNA transfer agent |
US7048925B2 (en) | 2003-08-28 | 2006-05-23 | Nitto Denko Corporation | Acid-sensitive polyacetals and methods |
DE602004032553D1 (en) * | 2003-09-05 | 2011-06-16 | Gen Hospital Corp | POLYACETAL DRUG CONJUGATES AS A RELEASE SYSTEM |
WO2005032597A1 (en) | 2003-09-29 | 2005-04-14 | Nitto Denko Corporation | Biodegradable polyacetals for in vivo polynucleotide delivery |
EP1682161A4 (en) * | 2003-10-29 | 2011-12-07 | Gentis Inc | Polymerizable emulsions for tissue engineering |
GB0329654D0 (en) * | 2003-12-23 | 2004-01-28 | Smith & Nephew | Tunable segmented polyacetal |
US7446099B2 (en) | 2004-02-27 | 2008-11-04 | Nitto Denko Corporation | Compositions and methods for biodegradable polymer-peptide mediated transfection |
DE102004023117A1 (en) * | 2004-05-11 | 2005-12-08 | A. u. K. Müller GmbH & Co KG | Solenoid valve for liquid media, especially hot water |
US7358223B2 (en) * | 2004-10-04 | 2008-04-15 | Nitto Denko Corporation | Biodegradable cationic polymers |
US7521415B2 (en) * | 2004-10-18 | 2009-04-21 | Nitto Denko Corporation | Methods of intracellular peptide delivery |
US7674452B2 (en) * | 2005-03-16 | 2010-03-09 | Nitto Denko Corporation | Polymer coating of cells |
EP1865989A2 (en) * | 2005-03-31 | 2007-12-19 | AP Pharma, Inc. | Peg-poly(ortho ester) graft copolymers and pharmaceutical compositions |
KR20070122521A (en) * | 2005-03-31 | 2007-12-31 | 에이피 파마, 인코포레이티드 | Peg-polyacetal and peg-polyacetal-poe graft copolymers and pharmaceutical compositions |
CN101495149A (en) * | 2005-03-31 | 2009-07-29 | 阿帕医药有限公司 | PEG-polyacetal diblock and triblock copolymers and pharmaceutical compositions |
US7588754B2 (en) * | 2005-05-10 | 2009-09-15 | Nitto Denko Corporation | Biodegradable polyacetals and methods |
US20060263328A1 (en) * | 2005-05-19 | 2006-11-23 | Sang Van | Hydrophilic polymers with pendant functional groups and method thereof |
JP2009504929A (en) * | 2005-08-18 | 2009-02-05 | スミス アンド ネフュー ピーエルシー | High-strength devices and composite materials |
EP1926506A2 (en) * | 2005-08-18 | 2008-06-04 | Smith & Nephew, PLC | Multimodal high strength devices and composites |
US7700541B2 (en) * | 2006-04-06 | 2010-04-20 | Nitto Denko Corporation | Biodegradable cationic polymers |
CA2679365C (en) | 2006-11-30 | 2016-05-03 | Smith & Nephew, Inc. | Fiber reinforced composite material |
EP2125775A4 (en) * | 2007-03-01 | 2011-07-13 | Cedars Sinai Medical Center | Antioxidant polymers containing [1,2]-dithiolane moieties and uses thereof |
WO2008129245A1 (en) | 2007-04-18 | 2008-10-30 | Smith & Nephew Plc | Expansion moulding of shape memory polymers |
AU2008243035B2 (en) | 2007-04-19 | 2013-09-12 | Smith & Nephew, Inc. | Graft fixation |
WO2008131197A1 (en) | 2007-04-19 | 2008-10-30 | Smith & Nephew, Inc. | Multi-modal shape memory polymers |
US8658148B2 (en) | 2007-06-22 | 2014-02-25 | Genzyme Corporation | Chemically modified dendrimers |
US9028874B2 (en) * | 2008-01-03 | 2015-05-12 | Cedars-Sinai Medical Center | Antioxidant nanosphere comprising [1,2]-dithiolane moieties |
EP2300451A1 (en) | 2008-06-02 | 2011-03-30 | Cedars-Sinai Medical Center | Nanometer-sized prodrugs of nsaids |
EP2370435B1 (en) | 2008-11-24 | 2015-01-07 | Cedars-Sinai Medical Center | Antioxidant camptothecin derivatives and antioxidant antineoplastic nanospheres thereof |
PE20100746A1 (en) * | 2008-12-10 | 2010-11-04 | Mersana Therapeutics Inc | PHARMACEUTICAL FORMULATIONS OF BIOCOMPATIBLE AND BIODEGRADABLE CAMPTOTECHIN-POLYMER CONJUGATES |
US8347887B2 (en) | 2008-12-23 | 2013-01-08 | Ams Research Corporation | Devices and methods for reversal of permanent sterilization |
WO2012129070A2 (en) | 2011-03-24 | 2012-09-27 | Unversity Of Florida Research Foundation, Inc. | Acetal metathesis polymerization |
EP2508544A1 (en) | 2011-04-04 | 2012-10-10 | Argon Pharma S.L. | Degradable polyacetal polymers |
US8815226B2 (en) | 2011-06-10 | 2014-08-26 | Mersana Therapeutics, Inc. | Protein-polymer-drug conjugates |
BR112013031819B1 (en) | 2011-06-10 | 2022-05-03 | Mersana Therapeutics, Inc | Polymeric support, pharmaceutical composition, compound and support use |
WO2014008375A1 (en) * | 2012-07-05 | 2014-01-09 | Mersana Therapeutics, Inc. | Terminally modified polymers and conjugates thereof |
CA2926586C (en) | 2013-10-11 | 2020-04-07 | Mersana Therapeutics, Inc. | Polymeric scaffold based on phf for targeted drug delivery |
KR102355745B1 (en) | 2013-10-11 | 2022-01-26 | 아사나 바이오사이언시스 엘엘씨 | Protein-polymer-drug conjugates |
EP3227348A4 (en) * | 2014-12-04 | 2018-07-18 | The Trustees of Columbia University in the City of New York | Biodegradable thermo-responsive polymers and uses thereof |
US11071714B2 (en) | 2016-04-29 | 2021-07-27 | Children's Medical Center Corporation | Poly(ketals) and related compositions and methods |
Family Cites Families (134)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US19446A (en) * | 1858-02-23 | Money in cars | ||
US135929A (en) * | 1873-02-18 | Improvement in the manufacture of car-springs | ||
US77295A (en) * | 1868-04-28 | theqdoee krausch | ||
US151668A (en) * | 1874-06-02 | Improvement in machines for cutting paper | ||
US466566A (en) * | 1892-01-05 | jenny | ||
US138474A (en) * | 1873-05-06 | Improvement in wave-powers | ||
US174411A (en) * | 1876-03-07 | Improvement in lamp-lighting devices | ||
US64050A (en) * | 1867-04-23 | Samuel jacob wallace | ||
US30444A (en) * | 1860-10-16 | william dopp | ||
US159A (en) * | 1837-03-31 | John swainson | ||
US90398A (en) * | 1869-05-25 | Improvement in tree-boxes | ||
US37300A (en) * | 1863-01-06 | Improvement in window-blind fastenings | ||
US54863A (en) * | 1866-05-22 | coghkan | ||
US115734A (en) * | 1871-06-06 | Improvement in brick-kilns | ||
US291668A (en) * | 1884-01-08 | Self-closing faucet | ||
US471036A (en) * | 1892-03-15 | Feeding mechanism for sewing-machines | ||
US419156A (en) * | 1890-01-07 | mcgiehan | ||
US176844A (en) * | 1876-05-02 | Improvement in beer-taps | ||
US152630A (en) * | 1874-06-30 | Improvement in cars for transporting coal | ||
US322392A (en) * | 1885-07-14 | Telepher | ||
US1482663A (en) * | 1922-01-12 | 1924-02-05 | Frederic A Shouldice | Stone gatherer |
US1579490A (en) * | 1924-10-16 | 1926-04-06 | Ke Haw Ke Mfg Company | Tube-vulcanizing device |
US2238477A (en) * | 1940-01-06 | 1941-04-15 | George E Murber | Golf rubber overshoe |
US2352547A (en) * | 1941-12-19 | 1944-06-27 | Pittsburgh Plate Glass Co | ?fining of tall oil |
US3268550A (en) | 1963-08-26 | 1966-08-23 | Ibm | Nu-butenyl carbazoles |
US4180064A (en) | 1972-12-27 | 1979-12-25 | Alza Corporation | System for delivering agent to environment of use over prolonged time |
US4014987A (en) | 1974-06-04 | 1977-03-29 | Alza Corporation | Device for delivery of useful agent |
US4093709A (en) | 1975-01-28 | 1978-06-06 | Alza Corporation | Drug delivery devices manufactured from poly(orthoesters) and poly(orthocarbonates) |
US4131648A (en) | 1975-01-28 | 1978-12-26 | Alza Corporation | Structured orthoester and orthocarbonate drug delivery devices |
US4180646A (en) | 1975-01-28 | 1979-12-25 | Alza Corporation | Novel orthoester polymers and orthocarbonate polymers |
US4079038A (en) | 1976-03-05 | 1978-03-14 | Alza Corporation | Poly(carbonates) |
US4152547A (en) * | 1977-11-29 | 1979-05-01 | Theis Peter F | Selective monitor for an automatic telephone answering system |
US4131662A (en) | 1978-01-03 | 1978-12-26 | Mobay Chemical Corporation | Talc-based external mold release agent for polyurethane foams |
US4261969A (en) | 1979-05-03 | 1981-04-14 | World Health Organization | Controlled drug release composition |
US4249531A (en) | 1979-07-05 | 1981-02-10 | Alza Corporation | Bioerodible system for delivering drug manufactured from poly(carboxylic acid) |
US4304767A (en) | 1980-05-15 | 1981-12-08 | Sri International | Polymers of di- (and higher functionality) ketene acetals and polyols |
US4502976A (en) | 1982-10-25 | 1985-03-05 | Bend Research, Inc. | Water soluble polyesters |
US4513143A (en) | 1982-12-01 | 1985-04-23 | Sri International | Preparation of ketene acetals |
US4690682A (en) | 1983-04-15 | 1987-09-01 | Damon Biotech, Inc. | Sustained release |
US4710497A (en) | 1983-05-20 | 1987-12-01 | Nitto Electric Industrial Co., Ltd. | Method for percutaneously administering physiologically active agents |
US5449670A (en) | 1983-05-20 | 1995-09-12 | Skinner; Wilfred A. | Composition and method for the transdermal delivery of bioactive peptides |
US4752612A (en) | 1983-07-01 | 1988-06-21 | Nitto Electrical Industrial Co., Ltd. | Method and percutaneously administering physiologically active agents using an alcohol adjuvant and a solvent |
US5128376A (en) | 1983-07-01 | 1992-07-07 | Nitto Denko Corporation | Method for percutaneously administering physiologically active agents using an adjuvant a solvent and a diol moderator |
US4590190A (en) | 1983-07-01 | 1986-05-20 | Nitto Electric Industrial Co., Ltd. | Method for percutaneously administering physiologically active agents using an alcohol adjuvant and a solvent |
US4548990A (en) | 1983-08-15 | 1985-10-22 | Ciba-Geigy Corporation | Crosslinked, porous polymers for controlled drug delivery |
US4818542A (en) | 1983-11-14 | 1989-04-04 | The University Of Kentucky Research Foundation | Porous microspheres for drug delivery and methods for making same |
US4605670A (en) | 1984-02-01 | 1986-08-12 | Nitto Electric Industrial Co., Ltd. | Method for percutaneously administering metoclopramide |
US4765973A (en) * | 1984-06-06 | 1988-08-23 | Merck & Co., Inc. | Polymers containing pendant acid functionalities and labile backbone bonds |
US4639366A (en) | 1984-06-06 | 1987-01-27 | Merck & Co., Inc. | Polymers containing pendant acid functionalities and labile backbone bonds |
GB8416234D0 (en) | 1984-06-26 | 1984-08-01 | Ici Plc | Biodegradable amphipathic copolymers |
US4549010A (en) | 1984-06-27 | 1985-10-22 | Merck & Co., Inc. | Bioerodible poly(ortho ester) thermoplastic elastomer from diketene diacetal |
GB8500209D0 (en) | 1985-01-04 | 1985-02-13 | Ceskoslovenska Akademie Ved | Synthetic polymeric drugs |
US4690825A (en) | 1985-10-04 | 1987-09-01 | Advanced Polymer Systems, Inc. | Method for delivering an active ingredient by controlled time release utilizing a novel delivery vehicle which can be prepared by a process utilizing the active ingredient as a porogen |
US4855132A (en) | 1986-02-25 | 1989-08-08 | S R I International | Method of preparing bioerodible polymers having pH sensitivity in the acid range and resulting product |
US4764364A (en) | 1986-02-25 | 1988-08-16 | S R I International | Method of preparing bioerodible polymers having pH sensitivity in the acid range and resulting product |
US4801457A (en) * | 1986-08-01 | 1989-01-31 | Sandoz Pharm. Corp. | Polyacetal hydrogels formed from divinyl ethers and polyols |
US4713441A (en) * | 1986-08-01 | 1987-12-15 | Sandoz Pharmaceuticals Corp. | Polyacetal hydrogels formed from divinyl ethers and polyols |
US4849426A (en) | 1987-05-15 | 1989-07-18 | Pearlman Dale L | Method for treating actinic keratosis with cytotoxic agents |
US5013553A (en) | 1987-06-30 | 1991-05-07 | Vipont Pharmaceutical, Inc. | Drug delivery devices |
US5080891A (en) | 1987-08-03 | 1992-01-14 | Ddi Pharmaceuticals, Inc. | Conjugates of superoxide dismutase coupled to high molecular weight polyalkylene glycols |
US4923645A (en) | 1987-11-16 | 1990-05-08 | Damon Biotech, Inc. | Sustained release of encapsulated molecules |
US4898928A (en) * | 1988-03-07 | 1990-02-06 | Sandoz Pharmaceuticals Corp. | Polyacetal and polyketal hydrogels formed from acetals or ketals and polyols |
US4957998A (en) | 1988-08-22 | 1990-09-18 | Pharmaceutical Delivery Systems, Inc. | Polymers containing acetal, carboxy-acetal, ortho ester and carboxyortho ester linkages |
US4938763B1 (en) | 1988-10-03 | 1995-07-04 | Atrix Lab Inc | Biodegradable in-situ forming implants and method of producing the same |
US5702716A (en) | 1988-10-03 | 1997-12-30 | Atrix Laboratories, Inc. | Polymeric compositions useful as controlled release implants |
US5725491A (en) | 1988-10-03 | 1998-03-10 | Atrix Laboratories, Inc. | Method of forming a biodegradable film dressing on tissue |
US5632727A (en) | 1988-10-03 | 1997-05-27 | Atrix Laboratories, Inc. | Biodegradable film dressing and method for its formation |
US5324520A (en) | 1988-12-19 | 1994-06-28 | Vipont Pharmaceutical, Inc. | Intragingival delivery systems for treatment of periodontal disease |
US4963369A (en) | 1989-01-19 | 1990-10-16 | Wm. Wrigley Jr. Co. | Gum composition containing dispersed porous beads containing active chewing gum ingredients and method |
US5108755A (en) | 1989-04-27 | 1992-04-28 | Sri International | Biodegradable composites for internal medical use |
EP0471036B2 (en) | 1989-05-04 | 2004-06-23 | Southern Research Institute | Encapsulation process |
US5028435A (en) | 1989-05-22 | 1991-07-02 | Advanced Polymer Systems, Inc. | System and method for transdermal drug delivery |
US4946931A (en) | 1989-06-14 | 1990-08-07 | Pharmaceutical Delivery Systems, Inc. | Polymers containing carboxy-ortho ester and ortho ester linkages |
US5013821A (en) | 1989-06-15 | 1991-05-07 | Pharmaceutical Delivery Systems, Inc. | Ortho and thio-ortho ester polymer |
US5077049A (en) | 1989-07-24 | 1991-12-31 | Vipont Pharmaceutical, Inc. | Biodegradable system for regenerating the periodontium |
US5487897A (en) | 1989-07-24 | 1996-01-30 | Atrix Laboratories, Inc. | Biodegradable implant precursor |
US5324519A (en) | 1989-07-24 | 1994-06-28 | Atrix Laboratories, Inc. | Biodegradable polymer composition |
US5030457A (en) | 1989-08-28 | 1991-07-09 | Pharmaceutical Delivery Systems, Inc. | Bioerodible polymers useful for the controlled release of therapeutic agents |
EP0489843A4 (en) | 1989-08-28 | 1992-08-12 | Pharmaceutical Delivery Systems, Inc. | Bioerodible polymers useful for the controlled release of therapeutic agents |
US5047464A (en) | 1989-09-20 | 1991-09-10 | Merck & Co., Inc. | Bioerodible thermoset elastomers |
US5217712A (en) | 1989-09-20 | 1993-06-08 | Merck & Co., Inc. | Bioerodible thermoset elastomers |
IL98087A (en) | 1990-05-04 | 1996-11-14 | Perio Prod Ltd | Colonic drug delivery system |
US5587507A (en) * | 1995-03-31 | 1996-12-24 | Rutgers, The State University | Synthesis of tyrosine derived diphenol monomers |
SE9002339L (en) | 1990-07-04 | 1992-01-05 | Kabi Pharmacia Ab | THERAPEUTIC COMPOSITION AND PROCEDURE FOR ITS PREPARATION |
NZ239370A (en) | 1990-08-22 | 1994-04-27 | Merck & Co Inc | Bioerodible implantable controlled release dosage form comprising a poly(ortho ester) or a polyacetal with an active agent incorporated into the chain backbone |
AU8935591A (en) | 1990-10-30 | 1992-05-26 | Alza Corporation | Drug delivery system and method |
US5422121A (en) | 1990-11-14 | 1995-06-06 | Rohm Gmbh | Oral dosage unit form |
HU222501B1 (en) * | 1991-06-28 | 2003-07-28 | Endorecherche Inc. | Controlled release pharmaceutical composition containing mpa or mga and process for its preparation |
US5211951A (en) * | 1991-07-24 | 1993-05-18 | Merck & Co., Inc. | Process for the manufacture of bioerodible poly (orthoester)s and polyacetals |
AU2605592A (en) | 1991-10-15 | 1993-04-22 | Atrix Laboratories, Inc. | Polymeric compositions useful as controlled release implants |
EP0560014A1 (en) | 1992-03-12 | 1993-09-15 | Atrix Laboratories, Inc. | Biodegradable film dressing and method for its formation |
WO1993019739A1 (en) | 1992-03-30 | 1993-10-14 | Alza Corporation | Viscous suspensions of controlled-release drug particles |
US5334640A (en) | 1992-04-08 | 1994-08-02 | Clover Consolidated, Ltd. | Ionically covalently crosslinked and crosslinkable biocompatible encapsulation compositions and methods |
US5461140A (en) | 1992-04-30 | 1995-10-24 | Pharmaceutical Delivery Systems | Bioerodible polymers for solid controlled release pharmaceutical compositions |
CA2134239C (en) * | 1992-06-09 | 2004-11-23 | Donald B. Axworthy | Pretargeting methods and compounds |
GB9217331D0 (en) | 1992-08-14 | 1992-09-30 | Xenova Ltd | Pharmaceutical compounds |
GB2270920B (en) | 1992-09-25 | 1997-04-02 | Univ Keele | Alginate-bioactive agent conjugates |
MY113268A (en) * | 1992-12-29 | 2002-01-31 | Insite Vision Incorporated | Plasticized bioerodible controlled delivery system |
GB2282384B8 (en) | 1993-08-18 | 1997-09-04 | Europ Economic Community | Drug delivery agents incorporating mitomycin |
IL110787A0 (en) * | 1993-08-27 | 1994-11-11 | Sandoz Ag | Biodegradable polymer, its preparation and pharmaceutical composition containing it |
US5681873A (en) | 1993-10-14 | 1997-10-28 | Atrix Laboratories, Inc. | Biodegradable polymeric composition |
CA2178964A1 (en) | 1993-12-15 | 1995-06-22 | John A. Duffy | Novel retinoid conjugate compounds and methods for treating skin aging |
GB9402809D0 (en) | 1994-02-14 | 1994-04-06 | Xenova Ltd | Pharmaceutical compounds |
GB9402805D0 (en) | 1994-02-14 | 1994-04-06 | Xenova Ltd | Pharmaceutical compounds |
IL112627A0 (en) | 1994-02-14 | 1995-05-26 | Xenova Ltd | Diketopiperazines, their preparation and pharmaceutical or veterinary compositions containing them |
CA2582666C (en) | 1994-04-08 | 2010-05-25 | Qlt Usa, Inc. | Controlled release implant |
US6007845A (en) | 1994-07-22 | 1999-12-28 | Massachusetts Institute Of Technology | Nanoparticles and microparticles of non-linear hydrophilic-hydrophobic multiblock copolymers |
US5607686A (en) | 1994-11-22 | 1997-03-04 | United States Surgical Corporation | Polymeric composition |
GB9426090D0 (en) | 1994-12-23 | 1995-02-22 | Xenova Ltd | Pharmaceutical compounds |
US5891877A (en) | 1995-02-14 | 1999-04-06 | Xenova Limited | Pharmaceutical compounds |
US5627187A (en) | 1995-04-12 | 1997-05-06 | Katz; Bruce E. | 5-FU for treating actinic kerotoses |
US5811510A (en) * | 1995-04-14 | 1998-09-22 | General Hospital Corporation | Biodegradable polyacetal polymers and methods for their formation and use |
US5877224A (en) | 1995-07-28 | 1999-03-02 | Rutgers, The State University Of New Jersey | Polymeric drug formulations |
US6096344A (en) * | 1995-07-28 | 2000-08-01 | Advanced Polymer Systems, Inc. | Bioerodible porous compositions |
US5916597A (en) | 1995-08-31 | 1999-06-29 | Alkermes Controlled Therapeutics, Inc. | Composition and method using solid-phase particles for sustained in vivo release of a biologically active agent |
US5968543A (en) | 1996-01-05 | 1999-10-19 | Advanced Polymer Systems, Inc. | Polymers with controlled physical state and bioerodibility |
IL118235A0 (en) * | 1996-05-13 | 1996-09-12 | Univ Ben Gurion | Composition and method for forming biodegradable implants in situ and uses of these implants |
US5993856A (en) | 1997-01-24 | 1999-11-30 | Femmepharma | Pharmaceutical preparations and methods for their administration |
US5965118A (en) | 1997-04-18 | 1999-10-12 | Access Pharmaceuticals, Inc. | Polymer-platinum compounds |
WO1998047496A2 (en) | 1997-04-18 | 1998-10-29 | Access Pharmaceuticals, Inc. | Polymer-platinum compounds |
JP2002511857A (en) * | 1997-06-11 | 2002-04-16 | ザ スクール オブ ファーマシー ユニヴァーシティ オブ ロンドン | Pharmaceutical compositions comprising an antibody-enzyme conjugate in combination with a prodrug |
US6338843B1 (en) * | 1997-06-12 | 2002-01-15 | Ml Laboratories | Biologically active materials |
US6585956B2 (en) * | 1997-07-07 | 2003-07-01 | The Dow Chemical Company | Dendritic-platinate drug delivery system |
US6193991B1 (en) * | 1997-10-29 | 2001-02-27 | Atul J. Shukla | Biodegradable delivery systems of biologically active substances |
US6048736A (en) | 1998-04-29 | 2000-04-11 | Kosak; Kenneth M. | Cyclodextrin polymers for carrying and releasing drugs |
US5939453A (en) | 1998-06-04 | 1999-08-17 | Advanced Polymer Systems, Inc. | PEG-POE, PEG-POE-PEG, and POE-PEG-POE block copolymers |
US6245345B1 (en) * | 1998-07-07 | 2001-06-12 | Atrix Laboratories, Inc. | Filamentous porous films and methods for producing the same |
US6261583B1 (en) * | 1998-07-28 | 2001-07-17 | Atrix Laboratories, Inc. | Moldable solid delivery system |
US6217895B1 (en) * | 1999-03-22 | 2001-04-17 | Control Delivery Systems | Method for treating and/or preventing retinal diseases with sustained release corticosteroids |
US6822086B1 (en) * | 1999-08-09 | 2004-11-23 | The General Hospital Corporation | Drug-carrier complexes and methods of use thereof |
EP1142596A1 (en) * | 2000-04-03 | 2001-10-10 | Universiteit Gent | Compositions of crosslinkable prepolymers for use in therapeutically active biodegradable implants |
AU2001288829A1 (en) * | 2000-09-06 | 2002-03-22 | Ap Pharma, Inc. | Degradable polyacetal polymers |
US6590059B2 (en) * | 2001-05-11 | 2003-07-08 | Ap Pharma, Inc. | Bioerodible polyorthoesters from dioxolane-based diketene acetals |
US6524606B1 (en) * | 2001-11-16 | 2003-02-25 | Ap Pharma, Inc. | Bioerodible polyorthoesters containing amine groups |
-
2001
- 2001-09-06 AU AU2001288829A patent/AU2001288829A1/en not_active Abandoned
- 2001-09-06 CA CA002453050A patent/CA2453050A1/en not_active Abandoned
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- 2001-09-06 EP EP01968589A patent/EP1315777B1/en not_active Expired - Lifetime
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- 2001-09-06 WO PCT/US2001/027664 patent/WO2002020663A2/en active IP Right Grant
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WO2002020663A2 (en) | 2002-03-14 |
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WO2002020663A3 (en) | 2002-05-10 |
DE60131177T2 (en) | 2008-08-07 |
EP1315777B1 (en) | 2007-10-31 |
US7220414B2 (en) | 2007-05-22 |
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