EP2173359A2 - Heparin-konjugate und verfahren - Google Patents
Heparin-konjugate und verfahrenInfo
- Publication number
- EP2173359A2 EP2173359A2 EP08778410A EP08778410A EP2173359A2 EP 2173359 A2 EP2173359 A2 EP 2173359A2 EP 08778410 A EP08778410 A EP 08778410A EP 08778410 A EP08778410 A EP 08778410A EP 2173359 A2 EP2173359 A2 EP 2173359A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- acid
- heparin
- residue
- composition
- bile acid
- 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.)
- Withdrawn
Links
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 title claims abstract description 157
- 229920000669 heparin Polymers 0.000 title claims abstract description 134
- 229960002897 heparin Drugs 0.000 title claims abstract description 121
- 238000000034 method Methods 0.000 title claims abstract description 50
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical group C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 claims abstract description 70
- 239000000203 mixture Substances 0.000 claims abstract description 61
- 239000003613 bile acid Substances 0.000 claims abstract description 40
- 125000006850 spacer group Chemical group 0.000 claims abstract description 32
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 31
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 21
- 201000011510 cancer Diseases 0.000 claims abstract description 16
- 239000003055 low molecular weight heparin Substances 0.000 claims description 35
- 229940127215 low-molecular weight heparin Drugs 0.000 claims description 35
- 230000000694 effects Effects 0.000 claims description 23
- WBWWGRHZICKQGZ-HZAMXZRMSA-N taurocholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCS(O)(=O)=O)C)[C@@]2(C)[C@@H](O)C1 WBWWGRHZICKQGZ-HZAMXZRMSA-N 0.000 claims description 23
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 claims description 20
- 125000003916 ethylene diamine group Chemical group 0.000 claims description 20
- 230000008685 targeting Effects 0.000 claims description 20
- WBWWGRHZICKQGZ-UHFFFAOYSA-N Taurocholic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(=O)NCCS(O)(=O)=O)C)C1(C)C(O)C2 WBWWGRHZICKQGZ-UHFFFAOYSA-N 0.000 claims description 17
- KXGVEGMKQFWNSR-LLQZFEROSA-N deoxycholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 KXGVEGMKQFWNSR-LLQZFEROSA-N 0.000 claims description 16
- 229960003964 deoxycholic acid Drugs 0.000 claims description 14
- 229920002971 Heparan sulfate Polymers 0.000 claims description 13
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical group C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 13
- RUDATBOHQWOJDD-UHFFFAOYSA-N (3beta,5beta,7alpha)-3,7-Dihydroxycholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 RUDATBOHQWOJDD-UHFFFAOYSA-N 0.000 claims description 12
- SMEROWZSTRWXGI-HVATVPOCSA-N lithocholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 SMEROWZSTRWXGI-HVATVPOCSA-N 0.000 claims description 11
- 235000019152 folic acid Nutrition 0.000 claims description 10
- 239000011724 folic acid Substances 0.000 claims description 10
- ZPBSAMLXSQCSOX-UHFFFAOYSA-K naphthalene-1,3,6-trisulfonate(3-) Chemical compound [O-]S(=O)(=O)C1=CC(S([O-])(=O)=O)=CC2=CC(S(=O)(=O)[O-])=CC=C21 ZPBSAMLXSQCSOX-UHFFFAOYSA-K 0.000 claims description 9
- 239000002202 Polyethylene glycol Substances 0.000 claims description 8
- 125000000217 alkyl group Chemical group 0.000 claims description 8
- 239000002634 heparin fragment Substances 0.000 claims description 8
- 229920001223 polyethylene glycol Polymers 0.000 claims description 8
- SMEROWZSTRWXGI-UHFFFAOYSA-N Lithocholsaeure Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 SMEROWZSTRWXGI-UHFFFAOYSA-N 0.000 claims description 7
- 229940014144 folate Drugs 0.000 claims description 7
- 150000004676 glycans Chemical class 0.000 claims description 7
- 229920001282 polysaccharide Polymers 0.000 claims description 7
- 239000005017 polysaccharide Substances 0.000 claims description 7
- 125000000472 sulfonyl group Chemical group *S(*)(=O)=O 0.000 claims description 7
- DKPMWHFRUGMUKF-UHFFFAOYSA-N (3alpha,5alpha,6alpha,7alpha)-3,6,7-Trihydroxycholan-24-oic acid Natural products OC1C(O)C2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 DKPMWHFRUGMUKF-UHFFFAOYSA-N 0.000 claims description 6
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 claims description 6
- JOYGXTIHTHBSOA-UHFFFAOYSA-N 1-(4-chlorophenyl)-3-thiophen-2-ylprop-2-en-1-one Chemical compound C1=CC(Cl)=CC=C1C(=O)C=CC1=CC=CS1 JOYGXTIHTHBSOA-UHFFFAOYSA-N 0.000 claims description 6
- OHXPGWPVLFPUSM-KLRNGDHRSA-N 3,7,12-trioxo-5beta-cholanic acid Chemical compound C1CC(=O)C[C@H]2CC(=O)[C@H]3[C@@H]4CC[C@H]([C@@H](CCC(O)=O)C)[C@@]4(C)C(=O)C[C@@H]3[C@]21C OHXPGWPVLFPUSM-KLRNGDHRSA-N 0.000 claims description 6
- 239000004380 Cholic acid Substances 0.000 claims description 6
- 108010015031 Glycochenodeoxycholic Acid Proteins 0.000 claims description 6
- 108010007979 Glycocholic Acid Proteins 0.000 claims description 6
- DGABKXLVXPYZII-UHFFFAOYSA-N Hyodeoxycholic acid Natural products C1C(O)C2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 DGABKXLVXPYZII-UHFFFAOYSA-N 0.000 claims description 6
- RFDAIACWWDREDC-UHFFFAOYSA-N Na salt-Glycocholic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(=O)NCC(O)=O)C)C1(C)C(O)C2 RFDAIACWWDREDC-UHFFFAOYSA-N 0.000 claims description 6
- RUDATBOHQWOJDD-BSWAIDMHSA-N chenodeoxycholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 RUDATBOHQWOJDD-BSWAIDMHSA-N 0.000 claims description 6
- 229960001091 chenodeoxycholic acid Drugs 0.000 claims description 6
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 claims description 6
- 229960002471 cholic acid Drugs 0.000 claims description 6
- 235000019416 cholic acid Nutrition 0.000 claims description 6
- 229960002997 dehydrocholic acid Drugs 0.000 claims description 6
- GHCZAUBVMUEKKP-GYPHWSFCSA-N glycochenodeoxycholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCC(O)=O)C)[C@@]2(C)CC1 GHCZAUBVMUEKKP-GYPHWSFCSA-N 0.000 claims description 6
- RFDAIACWWDREDC-FRVQLJSFSA-N glycocholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 RFDAIACWWDREDC-FRVQLJSFSA-N 0.000 claims description 6
- 229940099347 glycocholic acid Drugs 0.000 claims description 6
- DGABKXLVXPYZII-SIBKNCMHSA-N hyodeoxycholic acid Chemical compound C([C@H]1[C@@H](O)C2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 DGABKXLVXPYZII-SIBKNCMHSA-N 0.000 claims description 6
- RUDATBOHQWOJDD-DNMBCGTGSA-N isoursodeoxycholic acid Chemical compound C([C@H]1C[C@@H]2O)[C@@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 RUDATBOHQWOJDD-DNMBCGTGSA-N 0.000 claims description 6
- BHQCQFFYRZLCQQ-UTLSPDKDSA-N ursocholic acid Chemical compound C([C@H]1C[C@@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-UTLSPDKDSA-N 0.000 claims description 6
- RUDATBOHQWOJDD-UZVSRGJWSA-N ursodeoxycholic acid Chemical compound C([C@H]1C[C@@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 RUDATBOHQWOJDD-UZVSRGJWSA-N 0.000 claims description 6
- GHCZAUBVMUEKKP-UHFFFAOYSA-N ursodeoxycholic acid glycine-conjugate Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(=O)NCC(O)=O)C)C1(C)CC2 GHCZAUBVMUEKKP-UHFFFAOYSA-N 0.000 claims description 6
- 229960001661 ursodiol Drugs 0.000 claims description 6
- KXGVEGMKQFWNSR-DNZDVJRKSA-N 3alpha,12beta-Dihydroxy-5alpha-cholan-24-oic Acid Chemical compound C([C@@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@H](O)C1 KXGVEGMKQFWNSR-DNZDVJRKSA-N 0.000 claims description 5
- 125000003929 folic acid group Chemical group 0.000 claims description 4
- 150000002790 naphthalenes Chemical class 0.000 claims description 3
- 125000003219 lithocholic acid group Chemical group 0.000 claims 2
- 239000002253 acid Substances 0.000 claims 1
- 239000003858 bile acid conjugate Substances 0.000 abstract description 10
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 26
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 26
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 26
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 24
- ZMXDDKWLCZADIW-UHFFFAOYSA-N dimethylformamide Substances CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 17
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 16
- 239000000243 solution Substances 0.000 description 16
- 230000015572 biosynthetic process Effects 0.000 description 15
- 108020003175 receptors Proteins 0.000 description 15
- 102000005962 receptors Human genes 0.000 description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 15
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 14
- 238000003786 synthesis reaction Methods 0.000 description 14
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 10
- 241000699670 Mus sp. Species 0.000 description 10
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 8
- 239000002244 precipitate Substances 0.000 description 8
- 102000004207 Neuropilin-1 Human genes 0.000 description 7
- 108090000772 Neuropilin-1 Proteins 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 7
- 239000012153 distilled water Substances 0.000 description 7
- 238000011282 treatment Methods 0.000 description 7
- 230000004614 tumor growth Effects 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 108050006400 Cyclin Proteins 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- -1 Heparin Oligosaccharides Chemical class 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 6
- SJRJJKPEHAURKC-UHFFFAOYSA-N N-Methylmorpholine Chemical compound CN1CCOCC1 SJRJJKPEHAURKC-UHFFFAOYSA-N 0.000 description 6
- 102000009339 Proliferating Cell Nuclear Antigen Human genes 0.000 description 6
- 230000003213 activating effect Effects 0.000 description 6
- 229960000397 bevacizumab Drugs 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000011541 reaction mixture Substances 0.000 description 6
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 description 5
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 5
- 102100039894 Hemoglobin subunit delta Human genes 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 108010053099 Vascular Endothelial Growth Factor Receptor-2 Proteins 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
- 239000003102 growth factor Substances 0.000 description 5
- 239000002628 heparin derivative Substances 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 206010027476 Metastases Diseases 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 4
- 108091008605 VEGF receptors Proteins 0.000 description 4
- 102000009484 Vascular Endothelial Growth Factor Receptors Human genes 0.000 description 4
- 125000000539 amino acid group Chemical group 0.000 description 4
- 230000033115 angiogenesis Effects 0.000 description 4
- 230000010100 anticoagulation Effects 0.000 description 4
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 4
- 230000010261 cell growth Effects 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 230000003993 interaction Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000009401 metastasis Effects 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 101000808011 Homo sapiens Vascular endothelial growth factor A Proteins 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 3
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 3
- 238000005481 NMR spectroscopy Methods 0.000 description 3
- 238000013019 agitation Methods 0.000 description 3
- 238000002983 circular dichroism Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 238000006471 dimerization reaction Methods 0.000 description 3
- 229960000304 folic acid Drugs 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 102000058223 human VEGFA Human genes 0.000 description 3
- 238000003364 immunohistochemistry Methods 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 210000004088 microvessel Anatomy 0.000 description 3
- 230000002297 mitogenic effect Effects 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 230000014508 negative regulation of coagulation Effects 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- NXLNNXIXOYSCMB-UHFFFAOYSA-N (4-nitrophenyl) carbonochloridate Chemical compound [O-][N+](=O)C1=CC=C(OC(Cl)=O)C=C1 NXLNNXIXOYSCMB-UHFFFAOYSA-N 0.000 description 2
- ADFXKUOMJKEIND-UHFFFAOYSA-N 1,3-dicyclohexylurea Chemical compound C1CCCCC1NC(=O)NC1CCCCC1 ADFXKUOMJKEIND-UHFFFAOYSA-N 0.000 description 2
- 206010009944 Colon cancer Diseases 0.000 description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 2
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- APWFTHDYKJHNEV-NDEPHWFRSA-N NPC Chemical compound C1=C2C(CC)=C3CN(C(C4=C([C@@](C(=O)OC4)(O)CC)C=4)=O)C=4C3=NC2=CC=C1OC(=O)N1CCC(N)CC1 APWFTHDYKJHNEV-NDEPHWFRSA-N 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 150000001720 carbohydrates Chemical group 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 108010070321 cyclo(Arg-Gly-Asp-Tyr-Lys) Proteins 0.000 description 2
- 210000002889 endothelial cell Anatomy 0.000 description 2
- 210000002744 extracellular matrix Anatomy 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 239000012299 nitrogen atmosphere Substances 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 230000011664 signaling Effects 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 150000003431 steroids Chemical class 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L sulfate group Chemical group S(=O)(=O)([O-])[O-] QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- 229940086542 triethylamine Drugs 0.000 description 2
- 238000001291 vacuum drying Methods 0.000 description 2
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 1
- HCXJFMDOHDNDCC-UHFFFAOYSA-N 5-$l^{1}-oxidanyl-3,4-dihydropyrrol-2-one Chemical group O=C1CCC(=O)[N]1 HCXJFMDOHDNDCC-UHFFFAOYSA-N 0.000 description 1
- ILDXDBSWYJDHAL-UHFFFAOYSA-N 6-o-(2,5-dioxopyrrolidin-1-yl) 1-o-methyl hexanedioate Chemical compound COC(=O)CCCCC(=O)ON1C(=O)CCC1=O ILDXDBSWYJDHAL-UHFFFAOYSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 108091016585 CD44 antigen Proteins 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 206010052358 Colorectal cancer metastatic Diseases 0.000 description 1
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 1
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000009465 Growth Factor Receptors Human genes 0.000 description 1
- 108010009202 Growth Factor Receptors Proteins 0.000 description 1
- 102000008055 Heparan Sulfate Proteoglycans Human genes 0.000 description 1
- 238000012404 In vitro experiment Methods 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- 102000004317 Lyases Human genes 0.000 description 1
- 108090000856 Lyases Proteins 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 102000016611 Proteoglycans Human genes 0.000 description 1
- 108010067787 Proteoglycans Proteins 0.000 description 1
- 108090000054 Syndecan-2 Proteins 0.000 description 1
- SAQSTQBVENFSKT-UHFFFAOYSA-M TCA-sodium Chemical class [Na+].[O-]C(=O)C(Cl)(Cl)Cl SAQSTQBVENFSKT-UHFFFAOYSA-M 0.000 description 1
- 102000016549 Vascular Endothelial Growth Factor Receptor-2 Human genes 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000004037 angiogenesis inhibitor Substances 0.000 description 1
- 230000002491 angiogenic effect Effects 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000001772 anti-angiogenic effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 229940120638 avastin Drugs 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 125000002619 bicyclic group Chemical group 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 150000007942 carboxylates Chemical group 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000020411 cell activation Effects 0.000 description 1
- KXZJHVJKXJLBKO-UHFFFAOYSA-N chembl1408157 Chemical compound N=1C2=CC=CC=C2C(C(=O)O)=CC=1C1=CC=C(O)C=C1 KXZJHVJKXJLBKO-UHFFFAOYSA-N 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- RPKLZQLYODPWTM-KBMWBBLPSA-N cholanoic acid Chemical compound C1CC2CCCC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@@H](CCC(O)=O)C)[C@@]1(C)CC2 RPKLZQLYODPWTM-KBMWBBLPSA-N 0.000 description 1
- 238000001142 circular dichroism spectrum Methods 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 230000001268 conjugating effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 229960001760 dimethyl sulfoxide Drugs 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000002296 dynamic light scattering Methods 0.000 description 1
- 238000001493 electron microscopy Methods 0.000 description 1
- 238000002338 electrophoretic light scattering Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- UYMKPFRHYYNDTL-UHFFFAOYSA-N ethenamine Chemical class NC=C UYMKPFRHYYNDTL-UHFFFAOYSA-N 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000006193 liquid solution Substances 0.000 description 1
- 239000006194 liquid suspension Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 125000000636 p-nitrophenyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1*)[N+]([O-])=O 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 239000012925 reference material Substances 0.000 description 1
- 230000026267 regulation of growth Effects 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- JAJWGJBVLPIOOH-IZYKLYLVSA-M sodium taurocholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 JAJWGJBVLPIOOH-IZYKLYLVSA-M 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000019635 sulfation Effects 0.000 description 1
- 238000005670 sulfation reaction Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000007910 systemic administration Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- SOBHUZYZLFQYFK-UHFFFAOYSA-K trisodium;hydroxy-[[phosphonatomethyl(phosphonomethyl)amino]methyl]phosphinate Chemical compound [Na+].[Na+].[Na+].OP(O)(=O)CN(CP(O)([O-])=O)CP([O-])([O-])=O SOBHUZYZLFQYFK-UHFFFAOYSA-K 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 239000002525 vasculotropin inhibitor Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
- A61K31/727—Heparin; Heparan
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0075—Heparin; Heparan sulfate; Derivatives thereof, e.g. heparosan; Purification or extraction methods thereof
- C08B37/0081—Reaction with amino acids, peptides, or proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
-
- 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
- A61K47/54—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 compound
- A61K47/55—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 compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
- A61K47/551—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 compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid
-
- 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
- A61K47/54—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 compound
- A61K47/554—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 compound the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
-
- 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
-
- 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
- A61K47/66—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 the modifying agent being a pre-targeting system involving a peptide or protein for targeting specific cells
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L5/00—Compositions of polysaccharides or of their derivatives not provided for in groups C08L1/00 or C08L3/00
- C08L5/10—Heparin; Derivatives thereof
Definitions
- This invention relates to heparin conjugates and methods of making and using thereof. More particularly, this invention relates to heparin-bile acid conjugates, heparin-bile acid conjugates further including a targeting moiety, heparin-bile acid conjugates wherein the heparin is bonded to the bile acid through the 3-carbon of the bile acid, and heparin conjugates wherein heparin is covalently bonded to sulfonated moieties. Methods of using these conjugates for treating cancer are also described.
- Heparin has anti-tumoral and anti-inflammatory activities as well as its well known anti-coagulant activity.
- R. Sasisekharan et al. Roles of heparan-sulphate glycosa- minoglycans in cancer, 2 Nat. Rev. Cancer 521-528 (2002).
- growth factors which are key regulators for cell mitogenic activity.
- Growth factors usually bind with growth factor receptors and can modulate cell growth.
- vascular endothelial growth factor (VEGF) is a key protein in physiological angiogenesis (or neo-vascularization), or formation of new blood vessels.
- VEGF vascular endothelial growth factor
- Angiogenesis is a complex multi-step process involving endothelial cell activation, controlled proteolytic degradation of the extracellular matrix (ECM), proliferation and migration of endothelial cells, and formation of capillary vessel lumina.
- ECM extracellular matrix
- LMWH low molecular weight heparin
- MW 4500-6000 Da
- in vitro heparin fragments of less than 18 saccharide residues reduce activity of VEGF, and fragments of less than 10 saccharide residues inhibit activity of bFGF.
- G.C. Jayson et al. Heparin Oligosaccharides: Inhibitors of the Biological Activity of bFGF on Caco-2 Cells, 75 Br. J. Cancer 9-16 (1997); S.
- Be- vacizumab (Avastin®) is an FDA-approved, anti-angiogenic drug that is representative of such VEGF inhibitors.
- Bevacizumab is a basic monoclonal antibody that binds the negatively charged receptor binding domain of VEGF and, therefore, can block the interaction between VEGF and VEGF receptors (Flkl, KDR).
- L.M. Ellis Mechanisms of Action of Bevacizumab as a Component of Therapy for Metastatic Colorectal Cancer, 33 Semin. Oncol. S107 (2006); E. Bergsland & M.N. Dickler, Maximizing the Potential of Bevacizumab in Cancer Treatment, 9 Oncologist 36-42 (2004).
- LMWH can bind the heparin binding domain of VEGF.
- the sulfate groups of heparin can bind with positively charged amino acid residues, such as arginine, histidine, and lysine.
- a model of complexes formed between the heparin binding domain of VEGF and heparin or heparan sulfate predicts that sulfate and carboxylate groups of heparin contact these basic amino acid residues in the heparin-binding cleft of the VEGF protein. CJ.
- VEGF165-binding sites within heparan sulfate encompass two highly sulfated domains and can be liberated by K5 lyase, 281 J. Biol. Chem. 1731-1740 (2006).
- K5 lyase 281 J. Biol. Chem. 1731-1740 (2006).
- binding of heparin to VEGF results in an anti-angiogenic effect.
- Treatment of VEGF with either UFH or LMWH had no effect on tumor- associated angiogenesis in an experimental model of colon cancer metastasis in rat liver. S. M.
- VEGF comprises two main parts, a positively charged heparin binding domain
- HBD Vascular Endothelial Growth Factor Determinants for Binding KDR and FLT-I Receptors. Generation of Receptor- selective VEGF Variants by Site-directed Mutagenesis, 271 J. Biol. Chem. 5638-5646 (1996). Because HBD and RBD are in separate domains, even though heparin binds with HBD, the RBD maintains its structure.
- HBD-deleted VEGF can bind to the VEGF receptor, but mitogenic activity is absent.
- the HBD is critical to the cell growth activity of VEGF. RBD binding to VEGF cannot maintain cell growth.
- the VEGF receptor is a monomer before binding with the RBD of VEGF.
- This invention provides a composition represented by the formula:
- B is a bile acid residue or a bile acid analog
- H is a heparin residue
- S 1 is a first spacer
- S 2 is a second spacer
- T is a targeting moiety
- m is an integer of 0 to about 50
- n is an integer of 0 to about 10, with the proviso that both m and n cannot be 0, and wherein B and S 1 are bonded to each other through a 3-carbon of B, and S 1 and S 2 can be the same or different.
- m is about 1 to about 30.
- m is about 1 to about 10.
- n is 1 to about 10.
- This invention also provides a method of making a heparin-spacer-bile acid or heparin-spacer-bile acid analog conjugate and a method of making a heparin- spacer-bile acid-spacer-targeting moiety or heparin-spacer-bile acid analog- spacer-targeting moiety.
- This invention also provides a method of treating cancer comprising administering to an individual in need thereof an effective amount of a composition represented by the formula:
- B is a bile acid residue or a bile acid analog
- H is a heparin residue
- S 1 is a first spacer
- S 2 is a second spacer
- T is a targeting moiety
- m is an integer of 0 to about 50
- n is an integer of 0 to about 10, with the proviso that both m and n cannot be 0, and wherein B and S 1 are bonded to each other through a 3-carbon of B, and S 1 and S 2 can be the same or different.
- heparin-bile acid conjugates are available for treating cancer because they cna inhibit angiogenesis, metastasis and tumor growth.
- FIG. 1 illustrates a scheme for synthesis of taurocholic acid carbonate derivative
- FIG. 2 illustrates a scheme for synthesis of an ethylene amine derivative of taurocholic acid (Et-TCA) by reaction of CB-TCA with ethylene diamine such that the ethylene diamine replaces the 4-nitrophenyl group.
- Et-TCA ethylene amine derivative of taurocholic acid
- FIG. 3 illustrates a scheme for synthesis of a heparin-taurocholic acid conjugate by reaction of Et-TCA with heparin such that a carboxyl group on heparin bonds to the free amine group of Et-TCA, resulting in heparin conjugated to taurocholic acid through the 3-carbon thereof.
- FIG. 4 illustrates a scheme for synthesis of activated deoxycholic acid (aDOCA) from deoxycholic acid (DOCA).
- FIG. 5 illustrates a scheme for synthesis of JV-deoxycholylethylenediamine from aDOCA.
- FIG. 6 illustrates a scheme for synthesis of a heparin-DOCA conjugate from N- deoxycholylethylenediamine and heparin.
- FIG. 7 shows tumor volume as a function of days after inoculation of C3H/HeN mice with LMWH (5 mg/kg), HTlO (5 mg/kg), HT2 (5 mg/kg), and a control.
- FIG. 8 shows tumor volume as a function of days after inoculation of C3H/HeN mice with LMWH (0.5 mg/kg), HTlO (0.5 mg/kg), HTlO (1 mg/kg), HTlO (5 mg/kg), be- vacizumab (5 mg/kg), and a control.
- FIG. 9 shows tumor volume as a function of days after inoculation of C3H/HeN mice with HD (5 mg/kg), UFH (5 mg/kg), HL (5 mg/kg), and a saline control.
- FIG. 10 shows tumor volume as a function of days after inoculation of C3H/HeN mice with HD (1 mg/kg), HD (5 mg/kg), HD (10 mg/kg), heparin (5 mg/kg), and a saline control.
- FIG. 11 shows detection of microvessels and expression of the proliferating cell nuclear antigen (PCNA) in tumor tissues treated with a control, heparin, or HD, as detected by immunohistochemistry using either an anti-CD34 antibody or and anti- PCNA antibody.
- PCNA proliferating cell nuclear antigen
- FIG. 12-14 illustrates a scheme for synthesis of a cRGDyK-heparin-taurocholic acid conjugate.
- FIG. 15 illustrates a scheme for synthesis of a highly sulfated heparin derivative, namely, a heparin-naphthalene trisulfonic acid conjugate.
- An illustrative embodiment according to the present invention comprises a composition represented by the formula:
- B is a bile acid residue or a bile acid analog
- H is a heparin residue
- S 1 is a first spacer
- S 2 is a second spacer
- T is a targeting moiety
- m is an integer of 0 to about 50
- n is an integer of 0 to about 10, with the proviso that both m and n cannot be 0, and wherein B and S 1 are bonded to each other through a 3-carbon of B, and S 1 and S 2 can be the same or different.
- m is about 1 to about 30.
- m is about 1 to about 10.
- n is 1 to about 10.
- the bile acid residue may be selected, for example, from the group consisting of residues of cholic acid, deoxycholic acid, chenodeoxycholic acid, lithocholic acid, ursocholic acid, ursodeoxycholic acid, isoursodeoxycholic acid, lagodeoxycholic acid, glycocholic acid, taurocholic acid, glycodeoxycholic acid, glycochenodeoxycholic acid, dehydrocholic acid, hyocholic acid, and hyodeoxycholic acid.
- the heparin can comprise heparin of any type, such as unfractionated heparin, high molecular weight heparin, low molecular weight heparin, heparin fragments, recombinant heparin, heparin analogs, and sulfonated polysaccharides containing heparin activity, and the like.
- the spacer can comprise an alkyl chain, polyethylene glycol, an ethylene diamine residue, and the like.
- the targeting moiety may comprise a folic acid residue, a cRGD residue, and the like.
- An illustrative composition according to the present invention comprises a composition wherein B is a taurocholate residue, H is a low molecular weight heparin residue, S 1 and S 2 are an ethylene diamine residue, and T is a cRGD residue.
- Another illustrative composition according to the present invention comprises a composition wherein B is a lithocholate residue, H is a low molecular weight heparin residue, S 1 and S 2 are an ethylene diamine residue, and T is folate residue.
- Another illustrative embodiment according to the present invention comprises a composition comprising at least one sulfonated moiety, such as naphthalene trisulfonate, covalently bonded heparin.
- Other illustrative embodiments comprise compositions wherein analogs of naphthalene trisulfonate or sulfonated naphthalenes are substituted for naphthalene trisulfonate.
- Still another illustrative embodiment according to the present invention comprises a composition comprising heparin bonded to the 3-carbon of a bile acid or bile acid analog.
- the bile acid analog comprises a sulfonyl group.
- the composition may further comprise a spacer between heparin and the bile acid or bile acid analog, such as an ethylene diamine residue, an alkyl chain, polyethylene glycol, and the like.
- the composition may also further comprise a targeting moiety coupled to the heparin through a second spacer.
- Yet another illustrative embodiment of the invention comprises a method of treating cancer comprising administering to an individual in need thereof an effective amount of a composition represented by the formula:
- B is a bile acid residue or a bile acid analog
- H is a heparin residue
- S 1 is a first spacer
- S 2 is a second spacer
- T is a targeting moiety
- m is an integer of 0 to about 50
- n is an integer of 0 to about 10, with the proviso that both m and n cannot be 0, and wherein B and S 1 are bonded to each other through a 3-carbon of B, and S 1 and S 2 can be the same or different.
- Still another illustrative embodiment of the invention comprises a method of treating cancer comprising administering to an individual in need thereof an effective amount of a composition comprising heparin bonded to the 3-carbon of a bile acid or bile acid analog.
- Another illustrative embodiment of the invention comprises a method of making a heparin-spacer-bile acid or heparin-spacer-bile acid analog conjugate.
- the method comprises activating the 3-carbon of a bile acid or bile acid analog to result in an activated bile acid or activated bile analog, bonding a first spacer to the activated bile acid or activated bile acid analog to result in a spacer-bile acid or spacer-bile acid analog, activating a heparin to result in an activated heparin, and then bonding the activated heparin to the spacer-bile acid or spacer-bile acid analog to result in the heparin-spacer-bile acid or heparin-spacer-bile acid analog conjugate.
- Still another illustrative embodiment of the invention comprises a method of making a heparin-spacer-bile acid-spacer-targeting moiety or heparin-spacer-bile acid analog- spacer-targeting moiety conjugate.
- the method comprises activating the targeting moiety to result in an activated targeting moiety, bonding a second spacer to the activated targeting moiety to result in a second spacer-targeting moiety, and then bonding the second spacer-targeting moiety to a heparin-spacer-bile acid conjugate or heparin-spacer-bile acid analog conjugate to result in the heparin-spacer-bile acid- spacer-targeting moiety conjugate or heparin-spacer-bile acid analog-spacer-targeting moiety conjugate.
- Bile acids means natural and synthetic derivatives of the steroid, cholanic acid, including, without limitation, cholic acid, deoxycholic acid, chen- odeoxycholic acid, lithocholic acid, ursocholic acid, ursodeoxycholic acid, isours- odeoxycholic acid, lagodeoxycholic acid, glycocholic acid, taurocholic acid, gly- codeoxycholic acid, glycochenodeoxycholic acid, dehydrocholic acid, hyocholic acid, hyodeoxycholic acid, and mixtures thereof, and the like.
- Bile acid analogs can also be used according to the present invention. Examples of such bile acid analogs include bile acids bearing at least one sulfonyl group.
- ⁇ means an amount of a heparin conjugate that is nontoxic but sufficient to provide the desired effect and performance at a reasonable benefit/risk ratio attending any cancer treatment.
- administering means delivering the composition to the individual being treated such that the composition is capable of being circulated systemically to the parts of the body where cancer cells are located.
- the composition is preferably administered to the individual by systemic administration, typically by subcutaneous, intramuscular, or intravenous administration, or intraperitoneal administration.
- injectables for such use can be prepared in conventional forms, either as a liquid solution or suspension or in a solid form suitable for preparation as a solution or suspension in a liquid prior to injection, or as an emulsion.
- Suitable excipients include, for example, water, saline, dextrose, glycerol, ethanol, and the like; and if desired, minor amounts of auxiliary substances such as wetting or emulsifying agents, buffers, and the like can be added.
- An illustrative embodiment according to the present invention comprises a composition represented by the formula : [63] (B-SVH-(S 2 -T) n
- B is a bile acid residue or a bile acid analog
- H is a heparin residue
- S 1 is a first spacer
- S 2 is a second spacer
- T is a targeting moiety
- m is an integer of 0 to about 50
- n is an integer of 0 to about 10, with the proviso that both m and n cannot be 0, and wherein B and S 1 are bonded to each other through a 3-carbon of B, and S 1 and S 2 can be the same or different.
- n is about 1 to about 30, and even more typically m is about 1 to about
- n is about 1 to about 10.
- the heparin can comprise heparin of any type, such as unfractionated heparin, high molecular weight heparin, low molecular weight heparin, heparin fragment, recombinant heparin, heparin analogs, heparin sulfate, and sulfonated polysaccharides containing heparin activity, and the like.
- the spacers, S 1 and S 2 are independently selected from the group consisting of alkyl chains, polyethylene glycol, an ethylene diamine residue, and the like.
- the targeting moiety may comprise a folic acid residue, a cRGD residue, and the like.
- An illustrative composition according to the present invention comprises a composition wherein B is a taurocholate residue, H is a low molecular weight heparin residue, S 1 and S 2 are ethylene diamine residues, and T is a cRGD residue.
- Another illustrative composition according to the present invention comprise a composition wherein B is lithocholate residue, H is a low molecular weight heparin residue, S 1 and S 2 are ethylene diamine residues, and T is a folate residue.
- Another illustrative embodiment according to the present invention comprises a composition comprising at least one sulfonated moiety, such as naphthalene trisulfonate, covalently bonded to heparin.
- sulfonated moieties that can be substituted for naphthalene trisulfonate include analogs of naphthalene trisulfonate, other sulfonated naphthalenes, and the like.
- Still another illustrative embodiment according to the present invention comprises a composition comprising heparin bonded to the 3-carbon of a bile acid or bile acid analog.
- the bile acid analog comprises a sulfonyl group.
- the composition may further comprise a first spacer between heparin and the bile acid or bile acid analog, such as an ethylene diamine residue, an alkyl chain, polyethylene glycol, and the like.
- the composition may also further comprise a targeting moiety coupled to the heparin through a second spacer.
- Yet another illustrative embodiment of the invention comprises a method of treating cancer comprising administering to an individual in need thereof an effective amount of a composition represented by the formula:
- B is a bile acid residue or a bile acid analog
- H is a heparin residue
- S 1 is a first spacer
- S 2 is a second spacer
- T is a targeting moiety
- m is an integer of 0 to about 50
- n is an integer of 0 to about 10, with the proviso that both m and n cannot be 0, and wherein B and S 1 are bonded to each other through a 3-carbon of B, and S 1 and S 2 can be the same or different.
- Still another illustrative embodiment of the invention comprises a method of treating cancer comprising administering to an individual in need thereof an effective amount of a composition comprising heparin bonded to the 3-carbon of a bile acid or bile acid analog.
- Another illustrative embodiment of the invention comprises a method of making a heparin-spacer-bile acid or heparin-spacer-bile acid analog conjugate.
- the method comprises first activating the 3-carbon of a bile acid or bile acid analog to result in an activated bile acid or activated bile acid analog.
- a first spacer is bonded to the activated bile acid or activated bile acid analog to result in a spacer-bile acid or spacer- bile acid analog.
- Heparin is activated by reacting the heparin with an activating agent to result in an activated heparin.
- the activated heparin is bonded to the spacer- bile acid or spacer-bile acid analog to result in the heparin-spacer-bile acid or heparin- spacer-bile acid analog conjugate.
- Still another illustrative embodiment of the invention comprises a method of making a heparin-spacer-bile acid-spacer-targeting moiety or heparin-spacer-bile acid analog- spacer-targeting moiety conjugate.
- the method comprises reacting a targeting moiety with an activating agent to result in an activated targeting moiety.
- a second spacer is bonded to the activated targeting moiety to result in a second spacer-targeting moiety.
- the second spacer-targeting moiety is bonded to the previously described heparin-spacer-bile acid conjugate or heparin-spacer-bile acid analog conjugate to result in the heparin-spacer-bile acid-spacer-targeting moiety conjugate or heparin-spacer-bile acid analog-spacer-targeting moiety conjugate.
- TCA-LMWH Taurocholic acid
- TCA-TCA taurocholic acid ni- trophenyl carbonate
- DMF JV,./V-dimethylformamide
- NPC 4-nitrophenyl chloroformate
- Et-TCA an amine derivative of taurocholic acid
- FIG. 2 An amine derivative of taurocholic acid (Et-TCA) was synthesized (FIG. 2).
- CB-TCA 0.5 g was dissolved in 5 ml of DMF.
- 4-methyl-morpholine (0.144 g; Sigma) was added, followed by agitation for 1 h at 5O 0 C.
- This solution was slowly added to an excess of ethylenediamine, followed by agitation for 16 h at room tem ⁇ perature.
- the feed mole ratio of CT-TCA, 4-methylmorpholine, and ethylenediamine was 1:2:100. Since Et-TCA was water soluble, but ethylenediamine, 4-methylmorpholine, and NPC were not, acetone recrystallization was used to obtain pure Et-TCA, which was then dried under partial vacuum.
- heparin taurocholate HT
- Table 1 The coupling ratio and anticoagulant activity of each preparation were determined. Coupling ratio is expressed as the number of taurocholate moieties per heparin. Anticoagulant activity is expressed as the percent of activity of dialyzed LMWH, as determined with an FXa kit from Sigma.
- Heparin 100 mg was dissolved in 3 ml of formamide by gentle heating. Different amounts of EDC were mixed with heparin solutions at room temperature, followed by the addition of different amounts of LITHO-NH 2 dissolved in DMF with slight heating. The reaction was performed at room temperature for 24 h. The product was precipitated in excess cold acetone, and precipitates were stored at O 0 C, washed 5 times with cold acetone to remove unreacted LITHO-NH 2 , followed by drying under reduced pressure. The resulting precipitates were collected by lyophilization to give heparin- lithocholic acid derivatives.
- DOCA (Sigma) was mixed with DCC (7.4 g) and NHS (4.5 g) in 100 ml of tetrahydrofuran (THF). The mixture was reacted for 12 h at room temperature under a nitrogen atmosphere, then the precipitated dicyclohexylurea was removed by filtration. The filtrate was precipitated in n- hexane. The succinimido DOCA precipitate was filtered off and washed thoroughly with n-hexane, followed by vacuum drying at room temperature.
- aDOCA was converted to JV-deoxycholylethylenediamine according to the procedure illustrated in FIG. 5.
- JV-Deoxycholylethylenediamine (DOCA-NH 2 ) was synthesized by introducing ethylenediamine to the activated (with a succinimido group) DOCA.
- Succinimido DOCA (1 g) was dissolved in DMF (5 ml), and the solution was slowly added dropwise into ethylenediamine (13.4 ml) solution. After reaction for 6 h, the mixture was precipitated in distilled water.
- the white powder DOCA-NH 2 was obtained after washing 3 times with distilled water and drying at reduced pressure.
- heparin was conjugated to JV-deoxycholylethylenediamine according to the procedure illustrated in FIG. 6.
- Heparin 0.1 g was dissolved in formamide (5 ml) with gentle heating.
- Different amounts of EDC were mixed with heparin solutions at room temperature, followed by addition of different amounts of DOCA- NH 2 dissolved in DMF.
- the resulting solutions were stirred at room temperature under a nitrogen atmosphere for 24 h.
- precipitates were carefully washed 3 times with acetone to remove excess DOCA- NH 2 , followed by drying at reduced pressure.
- the dried HD conjugates were dissolved in water. Lyo- philization of the HD conjugates provided a white powder.
- heparin activity means the anticoagulation ability of heparin.
- COATEST HEPARIN FXa assay kit from Chromogenix was used for determining heparin activity of heparin conjugates. Results were recorded at 405 nm.
- the size of self-aggregated dispersions was determined using dynamic light scattering (Spectra Physics Laser Model 127-35) operated at 633 nm and 25+0.1 0 C. Scattered light was measured at an angle of 90° and collected with a BI-9000At auto- correlator. The concentration of HD conjugates was kept constant at 1 mg/ml. The hy- drodynamic diameter of self-aggregates was calculated by the Stokes-Einstein equation. The polydispersity factor, represented as 2 /T 2 was evaluated from the cumulant method, where 2 is the second cumulant of the decay function and F is the average characteristic line width. The zeta potentials of the nanoparticles were measured using an ELS- 8000 electrophoretic light scattering spectrophotometer (Otsuka Electronics Co., Ltd., Japan).
- the ultra-violet circular dichroism (CD) spectrum of proteins can predict important characteristics of their secondary structure. CD spectra can be readily used to estimate the fraction of a molecule that is in the alpha-helix conformation, the beta-sheet conformation, the beta-turn conformation, or some other (e.g., random coil) conformation. Jasco J-715 Circular Dichroism (Jasco, Japan) was used for making the determination.
- Table 2 shows the characterization of heparin derivatives prepared according to Examples 1-4.
- Example 6 [HO] Tumor Growth Inhibition [111] The anti-tumor activity of LMWH, HTlO, HT2, and control were compared in a tumor volume inhibition study. Seven-week-old male C3H/HeN mice (Orient Bio) were used for all animal experiments. Subcutaneous tumors were established by inoculating 1 x 10 6 SCC cells in the backs of the mice by subcutaneous injection. Care and maintenance of animals were performed in adherence to institutional guidelines of the Institutional Animal Care and Use Committee (IACUC).
- IACUC Institutional Animal Care and Use Committee
- mice When the tumors had grown to about 50-70 mm 3 , the mice were given intravenous injections of 0.1 ml of saline containing HTlO (5 mg/kg), HT2 (5 mg/kg), or LMWH (5 mg/kg), or saline alone (control), every three days. On the 15th day, the mice were sacrificed and their tumors removed. All treatment groups contained 7 or 8 mice. Tumor tissues were isolated from three representative treated and untreated tumor-bearing mice. Detection of micro vessels and expression of the proliferating cell nuclear antigen (PCNA) marker in tumor tissues were carried out by immunohistochemistry using a specific anti-CD34 antibody and anti-PCNA antibody (Dako, Carpinteria, California), respectively. FIG. 7 shows that HTlO resulted in about 69% tumor volume inhibition compared to the control treatment.
- PCNA proliferating cell nuclear antigen
- Example 6 The procedure of Example 6 was followed, except that HTlO (5 mg/kg), HTlO (1 mg/kg), HTlO (0.5 mg /kg), LMWH (0.5 mg/kg), and control were intravenously administered daily, and bevacizumab (5 mg/kg) was intraperitoneally administered twice weekly.
- HTlO is a nontoxic heparin derivative, and its anticoagulation activity is less than 13% that of LMWH, thus daily administration was not problematic.
- FIG. 8 shows that HTlO administered in doses of 1 mg/kg and 5 mg/kg was similar in its tumor growth inhibition effect to bevacizumab.
- HTlO (5 mg/kg) inhibited tumor growth about 71% compared with the control, while HTlO (1 mg/kg) inhibited tumor growth about 63%, and bevacizumab inhibited tumor growth about 74%.
- HTlO nor LMWH administered at 0.5 mg/kg were effective in inhibiting tumor growth.
- Example 6 The procedure of Example 6 was followed except that UFH (5 mg/kg), HL (5 mg/ kg), HD (5 mg/kg), and a control were administered. As shown in FIG. 9, HD and HL exhibited about 55-60% tumor volume inhibition compared with the control.
- HD was selected for a study on the in vivo dose effect of this heparin conjugate.
- the procedure of Example 6 was carried out, except that treatments were HD (10 mg/kg), HD (5mg/kg), HD (1 mg/kg), heparin (5 mg/kg), ad control.
- Detection of micro vessels and expression of PCNA was carried out using immunohistochemistry and anti-CD34 antibody and anti-PCNA antibody (Dako). Results are shown in FIGS. 10 and 11.
- FIGS. 12-14 show synthesis of a cRGD-HT conjugate.
- 50 mg of cRGDyK peptide was dissolved in DMF (10 ml) and then methyl-N-succinimidyl adipate (MSA, 32 mg) was added. After stirring for 12 h at room temperature, phenyl ester was added to the reaction mixture and stirring was continued for 6 h to protect the carboxyl group of the cRGDyK peptide.
- the reaction mixture was precipitated with a 3-fold excess of water to remove any unreacted reagents, and the precipitate was washed with water and then dried in vacuo.
- heparin in an amount indicated in column A was dissolved in the corresponding amount of formamide from column B, and this solution was agitated on an ice bath.
- EDAC in the corresponding amount from column C was added to the heparin solution.
- trisodium 8-amino-l,3,6-naphthalene trisulfonate (ANTS) in the corresponding amount from column D dissolved in the corresponding amount of formamide from column E was mixed with the activated heparin solution. After stirring for 12 h at room temperature, the reaction mixture was dialyzed against water using a 1000 MWCO membrane for 2 days and was then lyophilized.
- the amount of naphthalene trisulfonate in the resulting heparin derivative was determined by HPLC, and the anti-coagulation activity of this highly sulfated heparin derivative was evaluated using an anti-FXa chromogenic assay.
- Folate -heparin-lithocholate was prepared as follows. Folic acid (1 mmol) dissolved in 20 ml DMSO was reacted with DCC (1.2 mmol) and NHS (2 mmol) at 5O 0 C for 6 h. Folate has two " ⁇ - and ⁇ -carboxylic acids, but the ⁇ -carboxylic acid is more selectively activated due to its higher reactivity. The resulting folate-NHS was reacted with ethylene diamine (13 mmol) and pyridine (500 mg) at room temperature overnight.
- the folylethylamine (folate-NH 2 ) was precipitated by the addition of excess acetonitrile, and the precipitate was filtered and washed with diethyl ether before trying under vacuum to get yellow powder.
- This product was added to HL (100 mg), dissolved in 20 ml of formamide, and activated by EDAC (3.38 mg) with 5 ⁇ l of N, N - diisopropylethylamine (DIEA) for 12 h.
- the unreacted folate-NH 2 was removed by dialysis (MWCO 2000).
- the final product, FHL was obtained by lyophilization at a yield of 91%.
- the folate content in FHL was determined by quantitative UV spectrophotometry at 365 nm.
- the anti-coagulation activity of FHL was measured by Fxa chromogenic assay (COATEST®eparin, Milan, Italy).
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Nanotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Botany (AREA)
- Materials Engineering (AREA)
- Dermatology (AREA)
- Cell Biology (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Crystallography & Structural Chemistry (AREA)
- Biochemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US82459407A | 2007-06-29 | 2007-06-29 | |
PCT/KR2008/003731 WO2009005258A2 (en) | 2007-06-29 | 2008-06-27 | Heparin conjugates and methods |
Publications (1)
Publication Number | Publication Date |
---|---|
EP2173359A2 true EP2173359A2 (de) | 2010-04-14 |
Family
ID=40226647
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP08778410A Withdrawn EP2173359A2 (de) | 2007-06-29 | 2008-06-27 | Heparin-konjugate und verfahren |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP2173359A2 (de) |
KR (1) | KR101027161B1 (de) |
WO (1) | WO2009005258A2 (de) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102009028526A1 (de) * | 2009-08-13 | 2011-02-24 | Leibniz-Institut Für Polymerforschung Dresden E.V. | Verfahren zur Modifikation und Funktionalisierung von Sacchariden |
KR101313894B1 (ko) | 2011-04-08 | 2013-10-01 | 경북대학교 산학협력단 | 신규한 탈황산화된 헤파린-담즙산 유도체를 포함하는 염증성 질환의 예방 및 치료용 조성물 |
KR101702251B1 (ko) * | 2012-11-29 | 2017-02-02 | 에스티팜 주식회사 | 신규한 소포 수송에 이용하기 위한 담즙산 올리고머 결합체 및 이의 용도 |
KR20210122492A (ko) * | 2020-04-01 | 2021-10-12 | 에스티팜 주식회사 | 항암 요법을 위한 헤파린-담즙산 올리고머 컨쥬게이트 |
US12030912B2 (en) * | 2020-05-15 | 2024-07-09 | Brigham Young University | Cationic steroidal antimicrobial compounds with urethane linkages |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR100378109B1 (ko) * | 2000-10-24 | 2003-03-29 | 주식회사 메디프렉스 | 소수성 다중복합 헤파린 결합체, 그의 제조방법 및 용도 |
FR2864091B1 (fr) | 2003-12-19 | 2006-04-07 | Ethypharm Sa | Derive amphiphile d'heparine forme par couplage de l'heparine avec un acide biliaire |
-
2008
- 2008-06-27 KR KR1020080061613A patent/KR101027161B1/ko not_active IP Right Cessation
- 2008-06-27 WO PCT/KR2008/003731 patent/WO2009005258A2/en active Application Filing
- 2008-06-27 EP EP08778410A patent/EP2173359A2/de not_active Withdrawn
Non-Patent Citations (1)
Title |
---|
See references of WO2009005258A2 * |
Also Published As
Publication number | Publication date |
---|---|
WO2009005258A3 (en) | 2009-02-26 |
KR20090003128A (ko) | 2009-01-09 |
WO2009005258A2 (en) | 2009-01-08 |
KR101027161B1 (ko) | 2011-04-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US8088753B2 (en) | Heparin conjugates and methods | |
CN103751795B (zh) | 透明质酸‑抗肿瘤药偶联物及复合纳米粒组合物的制备和应用 | |
AU2006274992B2 (en) | Antitumoral bioconjugates of hyaluronic acid or its derivatives obtained by indirect chemical conjugation | |
JP4704753B2 (ja) | ヒアルロン酸又はヒアルロン酸誘導体に共有結合したタキサン類 | |
Pasut et al. | PEG-epirubicin conjugates with high drug loading | |
CN101396563B (zh) | 以奥曲肽为靶向配基的壳聚糖衍生物及其在药剂中的应用 | |
Cheewatanakornkool et al. | Thiolated pectin–doxorubicin conjugates: Synthesis, characterization and anticancer activity studies | |
CN108524919A (zh) | 用于改进的药物递送的载体 | |
JP2015505559A (ja) | 治療剤送達のためのシクロデキストリン系ポリマー | |
CA2257233A1 (en) | Process for producing drug complexes | |
WO2008136536A1 (ja) | 化学架橋ヒアルロン酸誘導体を含むハイブリッドゲルおよびそれを用いた医薬組成物 | |
Sargazi et al. | Hyaluronic acid/polyethylene glycol nanoparticles for controlled delivery of mitoxantrone | |
EP2173359A2 (de) | Heparin-konjugate und verfahren | |
CN112933219B (zh) | 一种dna-多肽可逆共价偶联分子的制备方法及其用途 | |
CN108794654A (zh) | 一种生物可降解的氧化还原敏感型聚合物及其制备方法和应用 | |
US20230066990A1 (en) | Hyaluronic acid derivative, pharmaceutical composition, and hyaluronic acid derivative-drug complex | |
KR101201473B1 (ko) | 자기 조립형 약물 및 세포 전달체의 제조 방법 및 이에 의해 제조된 자기 조립형 약물 및 세포 전달체 | |
Wang et al. | A novel biomimetic chitosan-based nanocarrier with suppression of the protein-nanocarrier interactions | |
Warrier et al. | Novel derivatives of arabinogalactan, pullulan & lactobionic acid for targeting asialoglycoprotein receptor: Biomolecular interaction, synthesis & evaluation | |
WO2009145594A2 (ko) | 약물전달체 | |
CN109908084A (zh) | 一种铂交联喜树碱前药胶束纳米药物及其制备方法和应用 | |
CN110177566A (zh) | 用于制备靶向肿瘤-脉管系统的抗肿瘤剂的方法 | |
US20090170195A1 (en) | Curcumin-hyaluronan compounds | |
JP5189243B2 (ja) | 薬物複合体および薬物送達用担体 | |
WO2022068927A1 (zh) | 具有肺靶向的肝素-多肽双重接枝环糊精骨架组合物及其制备方法与应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 20100129 |
|
AK | Designated contracting states |
Kind code of ref document: A2 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MT NL NO PL PT RO SE SI SK TR |
|
AX | Request for extension of the european patent |
Extension state: AL BA MK RS |
|
RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: PARK, RANG-WOON Inventor name: KIM, SANG YOON Inventor name: JEON, OK-CHEOL Inventor name: LEE, E SAK Inventor name: BYUN, YOUNGRO |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION HAS BEEN WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 20110601 |
|
18W | Application withdrawn |
Effective date: 20110601 |