AU8667791A - Inhibition of metastasis potential and invasiveness by oligosaccharides or oligosaccharide antigens or antibodies - Google Patents
Inhibition of metastasis potential and invasiveness by oligosaccharides or oligosaccharide antigens or antibodiesInfo
- Publication number
- AU8667791A AU8667791A AU86677/91A AU8667791A AU8667791A AU 8667791 A AU8667791 A AU 8667791A AU 86677/91 A AU86677/91 A AU 86677/91A AU 8667791 A AU8667791 A AU 8667791A AU 8667791 A AU8667791 A AU 8667791A
- Authority
- AU
- Australia
- Prior art keywords
- sialosyl
- oligosaccharide
- tumor
- group
- antigens
- 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.)
- Granted
Links
- 150000002482 oligosaccharides Chemical class 0.000 title claims description 54
- 229920001542 oligosaccharide Polymers 0.000 title claims description 52
- 239000000427 antigen Substances 0.000 title claims description 41
- 102000036639 antigens Human genes 0.000 title claims description 41
- 108091007433 antigens Proteins 0.000 title claims description 41
- 206010027476 Metastases Diseases 0.000 title claims description 39
- 230000009401 metastasis Effects 0.000 title claims description 33
- 230000005764 inhibitory process Effects 0.000 title description 9
- 206010028980 Neoplasm Diseases 0.000 claims description 32
- 210000004881 tumor cell Anatomy 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 26
- 150000001720 carbohydrates Chemical class 0.000 claims description 25
- 239000003795 chemical substances by application Substances 0.000 claims description 21
- -1 poly(ethylene glycol) Polymers 0.000 claims description 18
- 230000002401 inhibitory effect Effects 0.000 claims description 17
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 16
- 238000002360 preparation method Methods 0.000 claims description 14
- 241001465754 Metazoa Species 0.000 claims description 11
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 10
- 239000008101 lactose Substances 0.000 claims description 10
- 229920001223 polyethylene glycol Polymers 0.000 claims description 9
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 239000003814 drug Substances 0.000 claims description 3
- 210000004027 cell Anatomy 0.000 description 50
- FHNIYFZSHCGBPP-ABBMIVAOSA-N beta-D-Gal-(1->4)-beta-D-Glc-OMe Chemical compound O[C@@H]1[C@@H](O)[C@H](OC)O[C@H](CO)[C@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 FHNIYFZSHCGBPP-ABBMIVAOSA-N 0.000 description 24
- 206010061289 metastatic neoplasm Diseases 0.000 description 24
- 230000001394 metastastic effect Effects 0.000 description 23
- 210000004072 lung Anatomy 0.000 description 22
- 235000014633 carbohydrates Nutrition 0.000 description 14
- 238000002474 experimental method Methods 0.000 description 12
- 241000699670 Mus sp. Species 0.000 description 10
- 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 9
- 239000002953 phosphate buffered saline Substances 0.000 description 9
- 238000011282 treatment Methods 0.000 description 9
- 239000012980 RPMI-1640 medium Substances 0.000 description 8
- 210000002889 endothelial cell Anatomy 0.000 description 8
- 239000007924 injection Substances 0.000 description 8
- 238000002347 injection Methods 0.000 description 8
- 201000001441 melanoma Diseases 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- 238000000338 in vitro Methods 0.000 description 7
- 230000003993 interaction Effects 0.000 description 7
- 239000002502 liposome Substances 0.000 description 7
- 230000009467 reduction Effects 0.000 description 7
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 6
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 230000021164 cell adhesion Effects 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- 238000011580 nude mouse model Methods 0.000 description 5
- 235000000346 sugar Nutrition 0.000 description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 206010009944 Colon cancer Diseases 0.000 description 4
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 4
- 208000001382 Experimental Melanoma Diseases 0.000 description 4
- 241001529936 Murinae Species 0.000 description 4
- 241000699660 Mus musculus Species 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 230000015556 catabolic process Effects 0.000 description 4
- 239000006285 cell suspension Substances 0.000 description 4
- 208000029742 colonic neoplasm Diseases 0.000 description 4
- 238000006731 degradation reaction Methods 0.000 description 4
- 239000012894 fetal calf serum Substances 0.000 description 4
- 150000002339 glycosphingolipids Chemical class 0.000 description 4
- 210000004408 hybridoma Anatomy 0.000 description 4
- 238000002649 immunization Methods 0.000 description 4
- 230000003053 immunization Effects 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 125000005647 linker group Chemical group 0.000 description 4
- 210000003462 vein Anatomy 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- 102000005348 Neuraminidase Human genes 0.000 description 3
- 108010006232 Neuraminidase Proteins 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 3
- 210000002469 basement membrane Anatomy 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 239000003480 eluent Substances 0.000 description 3
- 150000002148 esters Chemical class 0.000 description 3
- 230000007717 exclusion Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 239000007928 intraperitoneal injection Substances 0.000 description 3
- 150000002597 lactoses Chemical class 0.000 description 3
- 238000004949 mass spectrometry Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 239000007790 solid phase Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- RMVRSNDYEFQCLF-UHFFFAOYSA-N thiophenol Chemical compound SC1=CC=CC=C1 RMVRSNDYEFQCLF-UHFFFAOYSA-N 0.000 description 3
- KFEUJDWYNGMDBV-UHFFFAOYSA-N (N-Acetyl)-glucosamin-4-beta-galaktosid Natural products OC1C(NC(=O)C)C(O)OC(CO)C1OC1C(O)C(O)C(O)C(CO)O1 KFEUJDWYNGMDBV-UHFFFAOYSA-N 0.000 description 2
- 108010024212 E-Selectin Proteins 0.000 description 2
- 102100023471 E-selectin Human genes 0.000 description 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical class OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- 102000016359 Fibronectins Human genes 0.000 description 2
- 108010067306 Fibronectins Proteins 0.000 description 2
- 229930186217 Glycolipid Natural products 0.000 description 2
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 2
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- KFEUJDWYNGMDBV-LODBTCKLSA-N N-acetyllactosamine Chemical compound O[C@@H]1[C@@H](NC(=O)C)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 KFEUJDWYNGMDBV-LODBTCKLSA-N 0.000 description 2
- HESSGHHCXGBPAJ-UHFFFAOYSA-N N-acetyllactosamine Natural products CC(=O)NC(C=O)C(O)C(C(O)CO)OC1OC(CO)C(O)C(O)C1O HESSGHHCXGBPAJ-UHFFFAOYSA-N 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 206010061535 Ovarian neoplasm Diseases 0.000 description 2
- 108010035766 P-Selectin Proteins 0.000 description 2
- 102100023472 P-selectin Human genes 0.000 description 2
- 206010035226 Plasma cell myeloma Diseases 0.000 description 2
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 2
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 239000013060 biological fluid Substances 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 2
- 238000010265 fast atom bombardment Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 230000009545 invasion Effects 0.000 description 2
- 201000005243 lung squamous cell carcinoma Diseases 0.000 description 2
- 210000001165 lymph node Anatomy 0.000 description 2
- 230000036210 malignancy Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000011987 methylation Effects 0.000 description 2
- 238000007069 methylation reaction Methods 0.000 description 2
- 201000000050 myeloid neoplasm Diseases 0.000 description 2
- 238000004393 prognosis Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 208000011581 secondary neoplasm Diseases 0.000 description 2
- 230000002269 spontaneous effect Effects 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 125000001424 substituent group Chemical group 0.000 description 2
- 150000008163 sugars Chemical class 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 230000035899 viability Effects 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 208000005443 Circulating Neoplastic Cells Diseases 0.000 description 1
- 241000193403 Clostridium Species 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 108700023372 Glycosyltransferases Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 238000005684 Liebig rearrangement reaction Methods 0.000 description 1
- NVGBPTNZLWRQSY-UWVGGRQHSA-N Lys-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(O)=O)CCCCN NVGBPTNZLWRQSY-UWVGGRQHSA-N 0.000 description 1
- 102000000440 Melanoma-associated antigen Human genes 0.000 description 1
- 108050008953 Melanoma-associated antigen Proteins 0.000 description 1
- 206010027480 Metastatic malignant melanoma Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 241000276498 Pollachius virens Species 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 1
- 102000019318 Selectin superfamily Human genes 0.000 description 1
- 108050006823 Selectin superfamily Proteins 0.000 description 1
- 102000003800 Selectins Human genes 0.000 description 1
- 108090000184 Selectins Proteins 0.000 description 1
- 102000009843 Thyroglobulin Human genes 0.000 description 1
- 108010034949 Thyroglobulin Proteins 0.000 description 1
- 229910021627 Tin(IV) chloride Inorganic materials 0.000 description 1
- 240000003834 Triticum spelta Species 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 206010064390 Tumour invasion Diseases 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 208000008385 Urogenital Neoplasms Diseases 0.000 description 1
- WOTQVEKSRLZRSX-JRFIZLOQSA-N [(2r,3r,4s,5r,6r)-4,5,6-triacetyloxy-3-[(2s,3r,4s,5s,6r)-3,4,5-triacetyloxy-6-(acetyloxymethyl)oxan-2-yl]oxyoxan-2-yl]methyl acetate Chemical compound CC(=O)OC[C@H]1O[C@H](OC(C)=O)[C@H](OC(C)=O)[C@@H](OC(C)=O)[C@@H]1O[C@H]1[C@H](OC(C)=O)[C@@H](OC(C)=O)[C@@H](OC(C)=O)[C@@H](COC(C)=O)O1 WOTQVEKSRLZRSX-JRFIZLOQSA-N 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 150000001266 acyl halides Chemical class 0.000 description 1
- 208000009956 adenocarcinoma Diseases 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 150000008064 anhydrides Chemical class 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 1
- 230000001588 bifunctional effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000001772 blood platelet Anatomy 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 230000009400 cancer invasion Effects 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 238000000262 chemical ionisation mass spectrometry Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000005757 colony formation Effects 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000002591 computed tomography Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000012631 diagnostic technique Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- CEIPQQODRKXDSB-UHFFFAOYSA-N ethyl 3-(6-hydroxynaphthalen-2-yl)-1H-indazole-5-carboximidate dihydrochloride Chemical compound Cl.Cl.C1=C(O)C=CC2=CC(C3=NNC4=CC=C(C=C43)C(=N)OCC)=CC=C21 CEIPQQODRKXDSB-UHFFFAOYSA-N 0.000 description 1
- 235000019439 ethyl acetate Nutrition 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 102000045442 glycosyltransferase activity proteins Human genes 0.000 description 1
- 108700014210 glycosyltransferase activity proteins Proteins 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 150000004820 halides Chemical class 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 235000020256 human milk Nutrition 0.000 description 1
- 210000004251 human milk Anatomy 0.000 description 1
- 210000004754 hybrid cell Anatomy 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 239000002955 immunomodulating agent Substances 0.000 description 1
- 230000001024 immunotherapeutic effect Effects 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 102000006495 integrins Human genes 0.000 description 1
- 108010044426 integrins Proteins 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 108010045069 keyhole-limpet hemocyanin Proteins 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 210000004880 lymph fluid Anatomy 0.000 description 1
- 230000001926 lymphatic effect Effects 0.000 description 1
- 108010054155 lysyllysine Proteins 0.000 description 1
- 229920001427 mPEG Polymers 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 208000021039 metastatic melanoma Diseases 0.000 description 1
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000000269 nucleophilic effect Effects 0.000 description 1
- 238000010534 nucleophilic substitution reaction Methods 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 210000003200 peritoneal cavity Anatomy 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 238000002135 phase contrast microscopy Methods 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 210000003281 pleural cavity Anatomy 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000011533 pre-incubation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- AAEVYOVXGOFMJO-UHFFFAOYSA-N prometryn Chemical compound CSC1=NC(NC(C)C)=NC(NC(C)C)=N1 AAEVYOVXGOFMJO-UHFFFAOYSA-N 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 210000001179 synovial fluid Anatomy 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000007910 systemic administration Methods 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 229960002175 thyroglobulin Drugs 0.000 description 1
- HPGGPRDJHPYFRM-UHFFFAOYSA-J tin(iv) chloride Chemical compound Cl[Sn](Cl)(Cl)Cl HPGGPRDJHPYFRM-UHFFFAOYSA-J 0.000 description 1
- FTVLMFQEYACZNP-UHFFFAOYSA-N trimethylsilyl trifluoromethanesulfonate Chemical compound C[Si](C)(C)OS(=O)(=O)C(F)(F)F FTVLMFQEYACZNP-UHFFFAOYSA-N 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 210000000264 venule Anatomy 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/001169—Tumor associated carbohydrates
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6093—Synthetic polymers, e.g. polyethyleneglycol [PEG], Polymers or copolymers of (D) glutamate and (D) lysine
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Oncology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Saccharide Compounds (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Description
Description
INHIBITION OF METASTASIS POTENTIAL AND INVASIVENES BY OLIGOSACCHARIDES OR OLIGOSACCHARIDE ANTIGENS OR ANTIBODIES
Technical Field
The present invention is generally directed toward the inhibition of tumor cell metastases and invasiveness, and more specifically, toward such inhibition through the use of agents including tumor-associated carbohydrate antigens and their oligosaccharide derivatives.
Background of the Invention
Despite enormous investments of financial and human resources, cancer remains one of the major causes of death. Current cancer therapies cure only about fifty percent of the patients who develop a malignant tumor. In most human malignancies, metastasis is the major cause of death.
Metastasis is the formation of a secondary tumor colony at a distant site. It is a multistep process of which tumor invasion is the first step. Tumor cells locally invade host tissue barriers, such as the epithelial basement membrane, to reach the interstitial stroma, where they gain access to blood vessels (or lymphatic channels) for further dissemination. After invading the endothelial layer of the vessel wall, the circulating tumor cells are dislodged into the circulation and arrest in the precapillary venules of the target organ by adherence to endothelial cell lumenal surfaces, or exposed basement membranes. The tumor cells again invade the vascular wall to enter the organ parenchyma. Finally, the extravasated tumor cell grows in a tissue different from where it originated.
In most human malignancies, distant metastases are often too small to be detected at the time the primary tumor is treated. Furthermore, widespread initiation of metastatic colonies usually occurs before clinical symptoms of metastatic disease are evident. The size and age variation in metastases, their dispersed anatomical location, and their heterogeneous composition are all factors that hinder surgical removal and limit the concentration of anticancer drugs that can be delivered to the metastatic colonies.
Due to the difficulties in the current approaches to the treatment and prevention of metastases, there is a need in the art for improved methods and
compositions for inhibiting metastasis potential of tumor cells. The present invention fills this need, and further provides other related advantages.
Summary of the Invention
Briefly stated, the present invention provides a variety of agents and methods for inhibiting metastasis potential and invasiveness of tumor cells. In one aspect of the present invention, a method for inhibiting tumor cell metastasis potential within a biological preparation is provided. The method comprises incubating the biological preparation with at least one agent selected from the group consisting of (a) tumor-associated carbohydrate antigens that exhibit differential prognostic significance, (b) antibodies that specifically bind to these antigens, (c) oligosaccharide components of these antigens, and (d) conjugates of these antigens or oligosaccharides, the agent inhibiting the metastasis potential of the preparation. Suitable biological preparations include cell cultures and biological fluids.
Another aspect of the present invention provides agents for use within the manufacture of a medicament for inhibiting metastasis potential of tumor cells in a warm-blooded animal. An agent is selected from the group consisting of (a) tumor-associated carbohydrate antigens that exhibit differential prognostic significance, (b) antibodies that specifically bind to these antigens, (c) oligosaccharide components of these antigens, and (d) conjugates of these antigens or oligosaccharides, wherein the agent is capable of inhibiting tumor cell metastasis potential.
Within a related aspect, the present invention provides a variety of glycoconjugates useful for prolonging the in vivo lifetime of oligosaccharides. The conjugates comprise an oligosaccharide coupled to poly(ethylene glycol).
Additional oligosaccharides for use within the present invention include lactose, lacto-N-tetrose, methyl β-D -lactoside and phenyl β-D- thiolactoside. Oligosaccharides may be used individually or in combination with one another.
These and other aspects of the present invention will become evident upon reference to the following detailed description and attached drawings. Brief Description of the Drawings
Figure 1 graphically illustrates survival of cancer patients with or without expression of a defined tumor associated carbohydrate antigen (TACA) in
their tumors. Panel A represents the expression of H/Ley/Leb antigen in lung squamous cell carcinoma as determined by MAb MIA-15-5. Panel B represents sialosyl-Lex expression in colonic cancer. Pane! C represents sialosyl-Tn expression in colonic cancer. Panel D represents sialosyl-Tn level in sera of ovarian cancer patients.
Figure 2 graphically illustrates the effects of methyl β -D-lactoside or phenyl β-D-thiolactoside on the number and size of lung colony deposits of BL6 cells. BL6 were preincubated with control medium, 0.1 M methyl β- D-lactoside ("Me-β-lactoside"), or 0.1 M phenyl β-D-thiolactoside ("phe-β-S-lactoside). 2 x 104 cells were injected intravenously into C57/BL mice. Lung colony numbers were counted at 21 days, and colonies were classified on the basis of diameter (> 1 mm vs. < 1 mm), as indicated for each column. Colony numbers are expressed per single lung. Number of experiments ("n") is indicated in parentheses.
Figure 3 graphically illustrates the effect of prior administration of methyl β-D-lactoside on the number and size of lung colony deposits of BL6 cells. Methyl β-D-lactoside (1 ml dosage) was injected intraperitoneally into C57/BL mice. After 10 minutes, B16 melanoma cells were injected intravenously. Lung colonies were counted and sized at 19 days. Group A represents control animals (not administered methyl β-D-lactoside) and groups B and C represent animals injected with 0.25 M and 0.5 M methyl β-D-lactoside, respectively. For each group, column 1 represents the total number of colonies, column 2 the number of colonies with diameter > 1 mm, and column 3 the number of colonies with diameter < 1 mm. Number of experiments is expressed as "n".
Figure 4 graphically illustrates the metastasis-inhibitory effect of methyl(Me)-β-lactoside. Tumor cells were intravenously injected, followed by intraperitoneal injection of: PBS control (A); 0.25 M Me-β-lactoside (B); 0.5 M Me-β-lactoside (C); 0.5 M lactose (D); 0.25 M N-acetyllactosamine (E); or 0.5 M Me-β-galactoside (E).
Figure 5 graphically illustrates that melanoma cell adhesion on LacCer is based on GM3-LacCer interaction. The order of metastatic potential is BL6>F10>F1 > >WA4. Panel A shows the order of melanoma cell adhesion on LacCer-coated solid phase. Panel B shows the order of melanoma cell adhesion on LacCer/Fibronectin (FN) co-coated solid phase. Panel C shows integrindependent adhesion.
Figure 6 graphically illustrates the melanoma cell (BL6) adhesion on LacCer (Panel A) and on endothelial cells (HUVEC) (Panel B) are inhibited by LacCer and GM3.
Figure 7 graphically illustrates H-Ley and H-H interaction. Panel A shows H1-liposome binding to various glycolipids. Panel B shows Ley-liposome binding to various glycolipids. Detailed Description of the Invention
As noted above, the present invention is directed towards agents and methods for the inhibition of tumor cell metastasis potential and invasiveness. Numerous tumor cells possess the ability to metastasize, i.e., to form a secondary tumor colony at a distant site. Sources of malignant tumor cells include melanoma, lung, breast, colorectal and urogenital cancers, such as bladder and prostate cancers. Within the present invention, the metastasis potential of tumor cells may be inhibited (i.e., inhibiting the ability of tumor cells to metastasize) through the use of (a) tumor-associated carbohydrate antigens (TACAs); (b) antibodies directed to these TACAs; (c) oligosaccharide components of these TACAs; or (d) conjugates of such TACAs, such as multivalent conjugates of lysyllysine or TACA-bearing glycosphingolipid (GSL) liposomes.
TACA epitopes play essential roles in tumor cell adhesion through their interaction with endothelial cells, platelets and basement membranes, whereby tumor metastasis and invasion may occur. The mechanism of adhesion may be based upon carbohydrate (CHO) - CHO interaction, CHO-lectin interaction or selectin family interaction. Adhesion of various tumor cells on activated endothelial cells and platelets is mediated primarily by the Leccam or selectin superfamily (e.g., ELAM-1, GMP-140). Colo205 tumor cells, which express type 1 chain sialosyl-Lea (SA-Lea) but not sialosyl-Lex (SA-Lex), adhere to endothelial cells. This adhesion was inhibited by anti-SA-Lea MAb, but not by SA-Lex MAb. These findings suggest that not only SA-Lex, but also SA-Lea, are the important ligands recognized by ELAM-1 and GMP-140.
Within the present invention, tumor metastasis and invasion is inhibited by blocking tumor cell adhesion, thereby significantly reducing or eliminating the spread of metastatic cells. TACAs suitable for use within the present invention are those showing differential prognostic significance (i.e., TACAs that may be clearly correlated with invasive or metastatic potential). Within the context of the present invention, such TACAs may be distinguished through a comparison of invasiveness, metastasis and clinical prognosis of similar tumors showing expression vs. non-expression of such TACAs. Preferred TACAs for use within the present invention include H/Ley/Leb, sialosyl-Lex (SA-Lex),
sialosyl-Lea (SA-Lea), and sialosyl-Tn (SA-Tn). Derivatives of such TACAs include dimeric Lex, sialosyl-dimeric Lex and trifuscosyl Lex .
The structures of sialosyl-Lex (structure 1), sialosyl-dimeric Lex (structure 2), dimeric Lex (Structure 3), trifucosyl L ex (structure 4), Leb (structure 5), H (structure 6), SA-Lea (structure 7), SA-Tn (structure 8), and GM3 (structure 9) are shown below.
Structure 1:
NeuAcα2→3Galβ1→4GlcNAcβ1-3Galβ1→4GlcNAcβ1→3Galβ1→4Glcβ1→R F
Structure 2:
NeuAcα2-3Galβ1→4GlcNAcβ1→3Galβ1→4GlcNAcβ1→3Galβ1→4Glcβ1→R
Structure 3:
Galβ1→4GlcNAcβ1→3Galβ1→4GlcNAcβ1→3Galβ1→4Glcβ1→R
Structure 4:
Galβ1→4GlcNAcβ1→3Galβ1→4GlcNAcβ1→3Galβ1→4Glcβ1→R
Structure 5:
Fucα1-2Galβ1→3GlcNAcβ1→3Galβ1→R
Structure 6:
Fucα1→2Galβ1→3GlcNAcβ1-3Galβ1→R
Structure 7:
NeuAcα2→3Galβ1→3GlcNAcβ1→3Galβ1→R
Structure 8:
NeuAcα2→6GalNAcα1→O-Ser/Thr
Structure 9:
NeuAcα2→3Galβ1→4Glcβ1→Cer
As noted above, TACAs for use within the present invention exhibit a differential prognostic significance. By way of example, such a differential prognostic significance may be illustrated by the fact that tumors expressing H/Ley/Leb antigens (as defined by MAb MIA-15-5) showed much worse patient prognosis than tumors not expressing these antigens. For instance, as shown in Figure 1 A, patients with squamous cell lung carcinoma expressing H/Ley/Leb had only an 11% survival over a 5-year period (i.e., 89% died), whereas comparable patients not expressing H/Ley/Le" had an approximately 62% survival over this period. Similar results were obtained for tumors showing expression vs. non- expression of SA-Lex and SA-Tn antigens. More specifically, as shown in Figure 1B, patients with colonic cancer expressing SA-Lex had only a 15% survival over a 5-year period, whereas comparable patients not expressing this antigen had an approximately 50% survival over this period. In a separate study, the 5-year survival of patients with early-stage colonic cancer not expressing SA-Tn was 100%, as compared to 75% for patients who expressed SA-Tn (see Figure 1C). As shown in Figure 1D, similar but more obvious differences were observed in patients with ovarian cancer showing expression vs. non-expression of SA-Tn antigen.
As noted above, antibodies to suitable TACAs may also be employed within the context of the present invention. As used herein, such antibodies include both monoclonal and polyclonal antibodies and may be intact molecules, a fragment of such a molecule, or a functional equivalent thereof. The antibody may be genetically engineered. Examples of antibody fragments include F(ab')2, Fab', Fab and Fv.
Briefly, polyclonal antibodies may be produced by immunization of an animal and subsequent collection of its sera. Immunization is accomplished, for example, by a systemic administration, such as by subcutaneous, intraspienic or intramuscular injection, into a rabbit, rat or mouse. It is generally preferred to follow the initial immunization with one or more booster immunizations prior to sera collection. Such methodology is well known and described in a number of references.
While polyclonal antibodies may be employed in the present invention, monoclonal antibodies (MAbs) are preferred. MAbs suitable within the present invention include those of murine or human origin, or chimeric antibodies such as those which combine portions of both human and murine antibodies (i.e., antigen binding region of murine antibody plus constant regions of human antibody). Human and chimeric antibodies may be produced using methods known by those skilled in the art Human antibodies and chimeric human-mouse antibodies are advantageous because they are less likely than murine antibodies to cause the production of anti-antibodies when administered clinically.
MAbs may be generally produced by the method of Kohler and Milstein (Nature 256:495-497, 1975; Eur. J. Immunol. 6:511-519, 1976), as well as by various techniques which modify their initial method (see Harlow and Lane (eds.), Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988, which is herein incorporated by reference in its entirety). Briefly, the lymph nodes and/or spleens of an animal immunized with one of the TACAs or their oligosaccharide components are fused with myeloma cells to form hybrid cell lines ("hybridomas" or "clones"). Each hybridoma secretes a single type of immunoglobulin and, like the myeloma cells, has the potential for indefinite cell division. It may be desirable to couple such molecules to a carrier to increase their immunogenicity. Suitable carriers include keyhole limpet hemocyanin, thyroglobulin, bovine serum albumin and derivatives thereof. An alternative to the production of MAbs via hybridomas is the creation of MAb expression libraries using bacteriophage and bacteria (e.g., Sastry et al., Proc. Natl. Acad. Sci USA 86:5728, 1989; Huse et al., Science 246:1275, 1989). Selection of antibodies exhibiting appropriate specificity may be performed in a variety of ways which will be evident to those skilled in the art
Representative examples of MAbs suitable for use within the present invention include MlA-15-5 (Miyake and Hakomori, Biochem. 30:3328,
1991), as well as the MAbs cited within Hakomori, Advances In Cancer Research 52:2 57-331, 1989.
As discussed above, oligosaccharide components of suitable TACAs may also be used within the present invention. As used herein, the term "oligosaccharides" includes naturally derived oligosaccharides, synthetically prepared, and derivatives of either, including portions of a TACA oligosaccharide component
Additional oligosaccharides useful within the present invention include lactose and lactose derivatives, such as methyl β-D -lactoside, lacto-N- tetrose (Galβ1→3GlcNAcβ1→3Galβ1→4Glc), and phenyl β-D-thiolactoside. Other lactose derivatives may also be used, including ethyl or phenyl lactoside and methyl or ethyl thiolactoside.
Other oligosaccharides suitable for inhibiting metastasis potential of cells of a particular tumor may be identified based upon determination of the structure of specific carbohydrate chain(s) which are involved in the tumor's ability to metastasize. The identification of carbohydrate-containing molecules involved in a tumor's ability to metastasize may be accomplished in a variety of ways, including through the use of glycosidases and inhibitors of glycosyltransferases. The structure of carbohydrates bound to either lipids or proteins may be determined based on degradation, mass spectrometry, including electron-impact direct-probe (El) and fast atom bombardment (FAB), and methylation analysis (techniques described, for example, in Nudelman et al., J. Biol. Chem. 261:5487-5495. 1986). Degradation analysis may be accomplished chemically and/or enzymatically, e.g., by glycosidases. The carbohydrate sequence suggested by degradation analysis may be determined by methylation analysis (Hakomori, J. Biochem. 55:205-208, 1964) followed by chemical ionization mass spectrometry of permethylated sugars (Stellner et al., Arch. Biochem. Biophys. 155:464-472, 1974; Levery et al., Meth. Enzymol. 138:13-25, 1987). Alternatively, or in conjunction with these techniques, El mass spectrometry may be performed on permethylated glycans or after the appropriate degradation of intact glycans (Kannagi et al., J. Biol. Chem. 259:8444-8451, 1984; Nudelman et al., J. Biol. Chem. 263:13942-13951, 1988). Homogeneity of the carbohydrate sequence may be demonstrated based on various chemical and physical criteria, including proton NMR spectroscopy of intact or methylated glycans and FAB mass spectrometry. Once the carbohydrate sequence has been determined, it will be evident to those of ordinary skill in the art to select an appropriate oligosaccharide for inhibiting the tumor cell's metastasis potential.
As briefly discussed above, conjugates of suitable TACAs or oligosaccharide components thereof, such as multivalent conjugates with lysyilysine or TACA-bearing giycosphingolipid (GSL) liposomes, may also be used within the present invention.
The components of the conjugate may be covalently coupled to one another either directly or via a linker group. A direct reaction between components is possible when each possesses a substituent capable of reacting with the other. For example, a nucleophilic group, such as an amino or sulfhydryl group, on one component may be capable of reacting with a carbonyl-containing group, such as an anhydride or an acyl halide, or with an alkyl group containing a good leaving group, e.g., a halide, on the other.
It may be desirable to covalently couple components via a linker group. It will be evident to those skilled in the art that a variety of bifunctional or polyfunctional reagents, both homo- and hetero-functional (such as those described in the Pierce Chemical Co. catalog, Rockford, IL), may be employed as the linker group. A linker group can serve to increase the chemical reactivity of a substituent and thus increase the coupling efficiency. An increase in chemical reactivity may also facilitate the use of functional groups on components which would not otherwise be possible. For example, a carboxyl group may be activated. Activation of a carboxyl group includes formation of an "active ester," such as a succinimidyl ester. The term "active ester" is known to refer to esters which are highly reactive in nucleophilic substitution reactions.
Alternatively, it may be desirable to produce conjugates in which the components are non-covalently linked. For example, one or more TACAs may be incorporated into the outer surface of GSL Iiposomes in a similar manner as previously described (Eggens et al., J. Biol. Chem. 264:9476-9484. 1989; Batzri and Korn, Biochim. Biophys. Acta 298:1015-1019, 1973; Szoka and Papahadjopoulos, Proc. Natl. Acad. Sci. USA 75:4194-4198. 1978).
It may be desirable to increase the in vivo lifetime of an oligosaccharide. As disclosed within the present invention, oligosaccharides may be coupled to (i.e., covalently bonded to) a straight-chain amphiphilic polymer, such as poly(ethylene glycol). A representative example of a method for producing an oligosaccharide-poly(ethylene glycol) conjugate is the reaction of an oligosaccharide, which has been derivatized to contain a succinimidyl group, with a poly(ethylene glycol) having a terminal amino group. The latter compound has a general formula of NH2-(CH2CH2-O)n-CH3, where n typically averages 44.7
(i.e., molecular weight of about 2,000) to 112.9 (i.e., molecular weight of about 5,000).
The inhibition of metastasis potential of tumor cells has a variety of in vitro and in vivo uses, e.g., treatment of isolated tumor cells or tumor-bearing hosts. Regarding in vitro aspects, as noted above, the present invention provides a method for inhibiting tumor cell metastasis potential within a biological preparation. The method comprises incubating a biological preparation with at least one agent selected from the group consisting of (a) tumor-associated carbohydrate antigens that exhibit differential prognostic significance, (b) antibodies that specifically bind to these antigens, (c) oligosaccharide components of these antigens, and (d) conjugates of these antigens or oligosaccharides, the agent inhibiting the metastasis potential of the preparation. Suitable biological preparations include cell cultures and cell suspensions in biological fluids, such as blood, urine, lymph, synovial and cerebrospinal fluid. TACAs, oligosaccharides or conjugates thereof will generally be incubated at a final concentration of about 0.1 to 1 M, and typically at about 02 to 03 M. Incubation is typically performed for 5 to 15 minutes at 37°C. After treatment of a biological preparation, the preparation may be injected or implanted in an animal, e.g., to confirm effectiveness of the inhibition of metastasis potential.
The present invention also provides uses of the agents in medicaments and methods for inhibiting tumor cell metastasis potential in a warm-blooded animal, such as a human. One or more agents is selected from the group consisting of (a) tumor-associated carbohydrate antigens that exhibit differential prognostic significance, (b) antibodies that specifically bind to these antigens, (c) oligosaccharide components of these antigens, and (d) conjugates of these antigens or oligosaccharides, the agent inhibiting the metastasis potential of the preparation. TACAs, oligosaccharides or conjugates thereof will generally be administered at a concentration of about 0.1 to 1 M and typically at about 0.2 to 03 M. It will be evident to those skilled in the art how to determine the optimal effective dose for a particular substance, e.g., based upon in vitro and in vivo studies in non-human animals. A variety of routes of administration may be used. Typically, administration will be intravenous or intracavitoiy, e.g., in pleural or peritoneal cavities, or in the bed of a resected tumor. A TACA, Ab, oligosaccharide or derivative as discussed above may be administered in combination with a pharmaceutically acceptable carrier or diluent, such as physiological saline. Moreover, such substances may be administered in combination with an immunotherapeutic or chemotherapeutic agent When a
combination of such a substance and an agent is desired, each compound may be administered sequentially, simultaneously, or combined and administered as a single composition. Diagnostic techniques, such as CAT scans, may be performed prior to and subsequent to administration to confirm the effectiveness of the inhibition of metastatic potential.
The following examples are offered by way of illustration and not by way of limitation.
EXAMPLES
Example 1
SYNTHESIS OF 1-ACΓOSE DERIVATIVES
A. Methyl β-D-lactoside
Heptaacetyllactosylimidate (Zimmeπnann et al., J. Carbohydr. Chem. 7:435, 1988) was reacted with methanol in dry dichloromethane containing trimethylsilyl trifluoromethanesulfonate according to standard procedure (Grundler and Schmit, Liebigs Ann. Chem. 1984:1826, 1984). Purification by silica-gel column chromatography (toluene/EtOAc 1:1), followed by de-O-acetylation with 0.01 M sodium mefhoxide, gave methyl β-D-lactoside in 68% yield from the imidate: m.p. 211-212°C (lit. 205°C, Smith and van Cleve, J. Am. Chem. Soc. 77:3159, 1955); [α]D + 13° (c 6.9, H2O) (lit + 1°, c 5.0, H2O), ibid).
B. Phenyl β-D-thiolactoside
Lactose octaacetate (Hudson and Kunz, J. Am. Chem. Soc. 47:2052, 1926) was treated with thiophenol and SnCl4 (Nicolaou et al., J. Am. Chem. Soc. 110:7910. 1988) in dichloromethane at 0°C to give phenyl heptaacetyl β-D-thiolactoside in 80% yield. This product was deacetylated with NaOMe in MeOH and neutralized with Amberlyst® 15. Purification of the product on a BioGel® P-2 column using water as an eluent, followed by lyophilization of the sugar-containing fraction, left phenyl β-D-thiolactoside as a white amorphous powder.
C. Lacto-N-tetrose
This oligosaccharide (Galβ1→3GlcNAcβ1→3Galβ1-4Glc) was prepared from human milk by pretreatment with ethanol and recycling BioGel P-2 column chromatography with water as eluent, followed by reversed-phase (C18) high pressure liquid chromatography with water (Dua and Bush, Anal. Biochem. 133:1. 1983). The 1H-NMR spectrum was superimposed on that of the authentic sample (BioCarb Chemicals, Lund, Sweden).
D. Poly(ethylene glycol) derivative of β-D-lactoside The reaction scheme is as follows:
The poly(ethylene glycol) derivative of β-D-lactoside was prepared from readily available 3-succinimidooxycarbonylpropyl O-(2, 3, 4, 6-tetra-O-acetyl- β-D-galactopyranosyl)-(1→4)-2,3,6-tri-O-acetyl-β-D-glucopyranoside (1) and poly(ethylene glycol) methyl ether (average M.W.2000; Aldrich Chemical, Milwaukee, WI) having terminal amino group (2) (Zalipsky et al., Eur. Polym. J. 19:1177, 1983). Treatment of 1 (100 mg, 0.12 mmol) and 2 (163 mg, 0.082 mmol) in dry N,N-dimethylformamide (2 mL) at room temperature for 2 hours gave, after chromatography on LH-20 with acetone as an eluent the β-D-lactoside heptaacetate 3 in 91% yield: [α]D -53° (c 0.5, chloroform). A subsequent saponification of 3 with 0.05 M sodium hydroxide at room temperature for one hour, followed by lyophilization, afforded the desired lactoside 4 quantitatively: [α]D -2.4° (c 1.0, chloroform).
Example 2
EFFECT OF LACTOSE AND LACTOSE DERIVATIVES ON METASTATIC POTENTIAL OF B16
MELANOMA CELLS
A. Cells and Animals
The highly metastatic BL6 clone of B16 melanoma cell line was obtained originally from Dr. Jean Starkey (Montana State Univ., Bozeman, MT), and clones were reselected in syngeneic C57/BL mice according to their metastatic potential. C57/BL mice were maintained in plastic cages under filtered air atmosphere and provided with water and food pellets ad lib. Cells were cultured in RPMI 1640 supplemented with 2 mM glutamine and 10% fetal calf serum (FCS), and detached with phosphate buffered saline (PBS) containing 2 mM EDTA. Viability was tested by trypan blue exclusion test
B. Effects of Oligosaccharides on Metastatic Potential
A suspension of BL6 cells (1-3 x 106 cells/ml RPMI 1640 medium) was prepared and aliquots were incubated in the presence or absence of various oligosaccharides at various concentrations, at 37°C for 5-10 minutes. Following incubation, typically, 3 x 104 or 2 x 104 cells (with or without oligosaccharide pretreatment) per 200 μl were injected via tail vein into 8-week-old female mice. After 18-21 days, mice were killed, lungs were fixed in 10% formaldehyde in PBS (pH 7.4), and tumor cell colonies were counted under a dissecting microscope, thus providing background values of metastatic melanoma colony number in lung under these conditions. Data on number and size of colonies were statistically
treated by the analysis of variance (ANOVA) procedure. Colonies with a diameter of 1 mm or greater were considered large-size and those with a diameter less than 1 mm were considered small-size.
For one experiment, BL6 cells were incubated with various concentrations of lactose, lacto-N-tetrose (Galβ1→3GlcNAcβ1→3Galβ1→4Glc), methyl β-D-lactoside, or phenyl β-D-thiolactoside for various durations. In the majority of experiments, a concentration of 0.1 M was used and cells were incubated at 37°C for 10 minutes, separated from sugar-containing medium by mild centrifugation at 400 x g for 10 minutes, resuspended in RPMI 1640, and injected (3 x 104 cells in 02 ml suspension) via tail vein. For some experiments, 2 x 104 cells were injected and colonies were counted at 21 days. Viability and cell growth ability of BL6 cells after incubation in various sugar solutions were tested by trypan blue exclusion test, and by plating in RPMI 1640 culture under normal conditions in vitro, as well as by subcutaneous inoculation in age-matched C57/BL mice in order to test tumor growth.
Lactose and lacto-N-tetrose showed 26% and 36% reductions, respectively, of metastatic colomes in lung when BL6 cells were preincubated with these sugars followed by intravenous injection of cells under identical conditions. Treatment of BL6 cells with 0.1 M, 0.01 M, or 0.005 M methyl β-D-lactoside under the same conditions as above resulted in (respectively) a 43%, 16%, and 8% reduction of metastatic lung colony number compared to control. The significant reduction caused by 0.1 M methyl β-D-lactoside was reproduced in three separate experiments and the reduction was found to be consistently between 35% and 45%.
In a second, completely independent series of experiments, treatment with methyl β-D-lactoside or phenyl β-D-thiolactoside under different conditions also produced a significant reduction of metastatic colonization, i.e., total colony number was reduced to 35% or 50% of control value following preincubation with methyl β-D-lactoside or phenyl β-D-thiolactoside re spectively. Reduction of larger-size colomes was more apparent than that of smaller colomes in all experiments, particularly those with phenyl β-D-thiolactoside (Figure 2). Methyl β -D-lactoside and phenyl β-D-thiolactoskc both showed a slight in vitro stimulatory effect on cell number increase and on thymidine incorporation. Thus, the inhibitory effect on tumor deposition is not related to the effect on cell growth in vitro or in vivo.
In a separate experiment, the effect of methyl β-D-lactoside on melanoma cell metastasis was determined after administration of
the oligosaccharide, followed by inoculation with tumor cells. Specifically, a one ml dosage of methyl β-D-lactoside (at a concentration of 0.25 M or 03 M) was injected intraperitoneaily in mice. After 10 minutes, B16 melanoma cells were injected intravenously. Lung colonies were counted 19 days later. Injection of methyl β-D-lactoside in advance of inoculation with tumor cells resulted in a significant reduction of lung metastatic colony formation (Figure 3).
In addition, the observations on the metastasis-inhibitory effect of methyl-β-lactoside have been extended to separate methyl-β-lactoside injection; i.e., tumor cells were intravenously injected, followed by intraperitoneal injection of methyl- β-lactoside. In these experiments, injection of 0.25-03 M methyl-β- lactoside reduced lung metastatic colony number by 40%-70% (see Figure 4; A=PBS control, B=0.25 M Me-β-lactoside; C=0.5 M Me-β-lactoside; D=0.5 M lactose; E=0.25 M N-acetyllactosamine; F=0.5 M Me-β-galactoside; intraperitoneal injection).
In a separate experiment, mouse melanoma B16 variants showing different degrees of metastatic potential (BL6/F10/F1/WA4) showed the same order of expression of GM3 ganglioside, which was previously identified as a melanoma-associated antigen (Hirabayashi et al., J. Biol. Chem. 260:13328. 1985; Nores et al., J. Immunol. 122:3171, 1987). GM3 interacts with LacCer, which is highly expressed on endothelial cells. The order of adhesion of the B16 variants onto LacCer-coated solid phase or onto endothelial cells was also in the same order as their metastatic potential. In contrast, integrin-dependent adhesion of the B16 variants was approximately equal for BL6, F10, and F1 (see Figure 5). These observations suggest that B16 adhesion of LacCer is based on molecular GM3-LacCer interaction. It has also been demonstrated that B16 melanoma adhesion on endothelial cells is inhibited not only by methyl-β-lactoside but also by LacCer liposome, Gg3Cer liposome, and GM3 liposome (see Figure 6).
Example 3
EXPRESSION OF SIALOSYL-DIMERIC LEX ON HUMAN LUNG ADENOCARONOMA CELL LINES AND
METASTATIC POTENTIAL
A. Cell lines
KUM-LK-2 is a human non-adenocarcinoma cell line characterized as producing spontaneous lung metastasis in nude mice. After screening 35 human carcinoma cell lines grown in nude mice, only this cell line produced metastatic deposits in nude mouse lung. KUM-LK-2 was used as the parent cell
line to obtain, by limiting dilution technique, sub-cell lines producing lung metastasis upon IV injection.
The procedure for the limiting dilution technique was as follows. KUM-LK-2 was cultured in RPMI 1640 medium (GIBCO, Grand Island, NY) supplemented with 10% FCS (Hyclone, Logan, UT) at 37°C in a 5% CO2/95% air atmosphere. Cells were treated briefly with 2 mM EDTA solution and washed twice with RPMI 1640 to make a single cell suspension in RPMI with 10% FCS. Cell viability was > 98% as determined by trypan blue exclusion staining. A cell suspension containing 1 cell per 100 μl was transferred to each well of a 96-well microtiter plate (Corning Glass Works, Corning, NY) and cultured continuously for 24 hours. Each well was then examined by phase contrast microscopy.
Three cell lines (HAL-8, HAL-24, and HAL-33) with different metastatic potential ("MP") were selected out of 25 clones obtained by limiting dilution technique on the basis of stable cell morphology. These 25 clones were originally selected from 63 clones showing stable morphology as well as consistent in vitro cell growth. All of these clones produced spontaneous lung metastasis. However, upon IV injection, clear differences were observed among the clones in terms of lung metastatic deposit formation. Two clones with high MP, five with low MP, and 18 with no MP were distinguished. Through repeated selection by IV injection of these clones, the most stable sub-cell lines showing consistent MP were established. These were HAL-8, -33, and -24, showing high, low, and no MP, respectively, to nu/nu mouse lung (see Table 1 below). Judging by macroscopic and microscopic examination, none of these three sub-cell lines showed metastasis in other organs or lymph nodes. The sub-cell lines represent stable variants originally present in KUM-LK-2. Based on chromosome analysis, these subclones are independent.
Nude mice were injected (2 x 105 cells) in the tail vein at various generation times as indicated. 56 days after injection, mice were killed and metastatic nodules on lung surface were counted under dissecting microscope. b Mean of 6 animals (range in parentheses). B. Expression of Cell Surface Carbohydrate Epitopes
The cell surface expression of various carbohydrate epitopes was analyzed by cytofluorometry using various monoclonal antibodies (MAbs) directed to Lex (MAb SH1), sialosyl-Lex (MAb SH4), sialosyl-dimeric Lex (MAb FH6), T (MAb HH8), Tn (MAb 1E3), and sialosyl-Tn (MAb TKH2). All antibodies used were culture supematants from their respective hybridomas, adjusted as 10 μg/ml of immunoglobulin.
Cells were detached from culture flask with 0.25% trypsin, 2 mM EDTA solution; 1 x 105 cells were prepared for each MAb treatment Cells were incubated with a MAb for 1 hour at 4°C and washed 2 times with RPMI 1640. Goat anti-mouse IgG or IgM-FTTC (Boehringer-Mannheim, Indianapolis, IN), diluted 50 times with PBS, was then added and incubated 30 minutes at 4°C. Finally, cells were washed 3 times, resuspended with PBS, and applied to an
EPICS PROFILE flow cytometiy (Epics, Hialeah, FL). These experiments were repeated with three different cell generations.
Patterns of expression of six carbohydrate epitopes (defined by their respective MAbs) on sub-cell lines HAL-8, -24, and -33 showed nearly identical profiles (as did the protein profiles for the three sub-cell lines), except in the case of sialosyl-dimeric Lex. In particular, HAL-8, -24, and -33 were found to highly and equally express sialosyl-Lex and sialosyl-Tn structures. Each of the three lines expressed low quantities of Lex and Tn, and did not express T. In contrast, expression of sialosyl-dimeric Lex was high on HAL-8, moderate on HAL-33, and low on HAL-24.
C. Inhibition of Metastasis by Sialidase Treatment of Cells
Cells were detached using 2 mM EDTA in PBS, washed, and resuspended in 9 volumes of PBS. One ml of cell suspension was incubated 5 minutes at 37°C with 0.2 U/ml of Clostridium perftingens sialidase (type X, Sigma Chemical Co., St Louis, MO). After incubation, cells were washed three times, resuspended with RPMI 1640, and investigated for MP and expression of sialosyl- dimeric Lex. MP of HAL-8 and -33 was completely inhibited by sialidase treatment of cells (see Table 2 below). Expression of sialosyl-dimeric Lex appears to play an important role in blood-borne metastasis.
a Nude mice were injected (2 x 105 cells) in the tail vein. 56 days after injection, mice were killed and metastatic nodules on lung surface were counted under dissecting microscope.
b Mean of 6 animals (range in parentheses).
From the foregoing, it will be evident that although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention.
Claims (13)
1. An agent selected from the group consisting of tumor-associated carbohydrate antigens that exhibit differential prognostic significance, antibodies that specifically bind to said antigens, oligosaccharide components of said antigens, and conjugates of said antigens or said oligosaccharides for use within the manufacture of a medicament for inhibiting tumor cell metastasis potential in a warm-blooded animal.
2. The agent of claim 1 wherein the oligosaccharide component is selected from the group consisting of lactose, lacto-N-tetrose, methyl D-lactoside, and phenyl D-thiolactoside.
3. The agent of claim 1 wherein the tumor-associated carbohydrate antigen is selected from the group consisting of sialosyl-Lex, H/Ley/Leb, sialosyl-Lea, sialosyl-Tn, dimeric Lex, sialosyl-dimeric Lex, and trifucosyl Lex.
4. The agent of claim 1 wherein the oligosaccharide component is the oligosaccharide group of a tumor-associated carbohydrate antigen selected from the group consisting of H/Ley/Leb, sialosyl-Lex, sialosyl-Lea, sialosyl-Tn, dimeric Lex, sialosyl-dimeric Lex, and trifucosyl Lex.
5. The agent of either of claims 2 or 4 wherein the oligosaccharide is coupled to poly(ethylene glycol).
6. A conjugate comprising an oligosaccharide coupled to poly(ethylene glycol).
7. The conjugate of claim 6 wherein the oligosaccharide is selected from the group consisting of lactose, lacto-N-tetrose, methyl D -lactoside, and phenyl D- thiolactoside.
8. The conjugate of claim 6 wherein the oligosaccharide is the oligosaccharide portion of a tumor-associated carbohydrate antigen selected from the group consisting of sialosyl-Lex, H/Ley/Leb, sialosyl-Lea, sialosyl-Tn, dimeric Lex, sialosyl-dimeric Lex, and trifucosyl Lex.
9. A method for inhibiting tumor cell metastasis potential within a biological preparation, comprising:
incubating the biological preparation with at least one agent selected from the group consisting of tumor-associated carbohydrate antigens that exhibit differential prognostic significance, antibodies that specifically bind to said antigens, oligosaccharide components of said antigens, and conjugates of said antigens or said oligosaccharides, said agent inhibiting the metastasis potential of the preparation.
10. The method of claim 9 wherein the oligosaccharide component is selected from the group consisting of lactose, lacto-N-tetrose, methyl D-lactoside, and phenyl D-thiolactoside.
11. The method of claim 9 wherein the tumor-associated carbohydrate antigen is selected from the group consisting of sialosyl-Lex, H/Ley/Leb, sialosyl-Lea, sialosyl-Tn, dimeric Lex, sialosyl-dimeric Lex, and trifucosyl Lex.
12. The method of claim 9 wherein the oligosaccharide component is the oligosaccharide group of a tumor-associated carbohydrate antigen selected from the group consisting of H/Ley/Leb, sialosyl-Lex , sialosyl-Lea, sialosyl-Tn, dimeric Lex, sialosyl-dimeric Lex, and trifucosyl Lex.
13. The method of claims 10 or 12 wherein the oligosaccharide is coupled to poly(ethylene glycol).
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US57553990A | 1990-08-30 | 1990-08-30 | |
US575539 | 1990-08-30 | ||
US72498391A | 1991-07-02 | 1991-07-02 | |
US724983 | 1991-07-02 | ||
PCT/US1991/006202 WO1992004048A2 (en) | 1990-08-30 | 1991-08-29 | Inhibition of metastasis potential and invasiveness by oligosaccharides or oligosaccharide antigens or antibodies |
Publications (2)
Publication Number | Publication Date |
---|---|
AU8667791A true AU8667791A (en) | 1992-03-30 |
AU659808B2 AU659808B2 (en) | 1995-06-01 |
Family
ID=27076723
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU86677/91A Ceased AU659808B2 (en) | 1990-08-30 | 1991-08-29 | Inhibition of metastasis potential and invasiveness by oligosaccharides or oligosaccharide antigens or antibodies |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP0546122A1 (en) |
JP (1) | JPH06501008A (en) |
AU (1) | AU659808B2 (en) |
CA (1) | CA2089375A1 (en) |
WO (1) | WO1992004048A2 (en) |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5162430A (en) * | 1988-11-21 | 1992-11-10 | Collagen Corporation | Collagen-polymer conjugates |
WO1993000588A1 (en) * | 1991-06-26 | 1993-01-07 | The Biomembrane Institute | Early diagnosis of lung cancer using anti-carbohydrate antibody combinations |
US5639737A (en) * | 1991-11-04 | 1997-06-17 | Co Enzyme Technology Ltd. | Method and compositions for treating malignant tumors and inhibiting growth and metastases of malignant tumors |
WO1994001483A1 (en) * | 1992-07-02 | 1994-01-20 | Collagen Corporation | Biocompatible polymer conjugates |
DE4236237A1 (en) * | 1992-10-27 | 1994-04-28 | Behringwerke Ag | Prodrugs, their preparation and use as medicines |
AU5289096A (en) * | 1995-04-14 | 1996-10-30 | Kazunori Kataoka | Polyethylene oxides having saccharide residue at one end and different functional group at another end, and process for producing the same |
US6841543B1 (en) | 1996-01-31 | 2005-01-11 | President And Fellows Of Harvard College | Methods of inhibiting production of T helper type 2 cytokines in human immune cells |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE3228233A1 (en) * | 1982-07-28 | 1984-02-02 | Wolfgang Dr. 6500 Mainz Dippold | The use of the antibody R-24 and method for the inhibition of the cell growth of melanoma cells |
US4904596A (en) * | 1985-08-07 | 1990-02-27 | Fred Hutchinson Cancer Research Center | Hybridoma antibody (FH6) defining a human cancer-associated difucoganglioside |
ES2058070T3 (en) * | 1986-05-09 | 1994-11-01 | Pulverer Gerhard | USE OF SPECIFIC MONOSACCHARIDES FOR THE PREPARATION OF A MEDICINAL PRODUCT TO PREVENT METASTASIS OF MALIGNANT TUMORS. |
EP0380084A3 (en) * | 1989-01-27 | 1991-03-13 | The Biomembrane Institute | Method for the inhibition of cell-cell and cell substratum interactions by the blocking of carbohydrate-carbohydrate interactions |
-
1991
- 1991-08-29 AU AU86677/91A patent/AU659808B2/en not_active Ceased
- 1991-08-29 EP EP91919388A patent/EP0546122A1/en not_active Withdrawn
- 1991-08-29 WO PCT/US1991/006202 patent/WO1992004048A2/en not_active Application Discontinuation
- 1991-08-29 CA CA002089375A patent/CA2089375A1/en not_active Abandoned
- 1991-08-29 JP JP3516668A patent/JPH06501008A/en active Pending
Also Published As
Publication number | Publication date |
---|---|
JPH06501008A (en) | 1994-01-27 |
EP0546122A1 (en) | 1993-06-16 |
WO1992004048A2 (en) | 1992-03-19 |
AU659808B2 (en) | 1995-06-01 |
WO1992004048A3 (en) | 1992-10-01 |
CA2089375A1 (en) | 1992-03-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US6121233A (en) | Methods for the inhibition of cancer metastasis mediated by endothelial adhesion molecules | |
EP0580763B1 (en) | Compositions and methods for endothelial binding | |
US5240833A (en) | Method for the production of monoclonal antibodies directed to tumor-associated gangliosides and fucogangliosides | |
CA1337403C (en) | Methods for the production of antibodies and induction of immune responses to tumor-associated gangliosides by immunization with ganglioside lactones | |
Casadesús et al. | A shift from N-glycolyl-to N-acetyl-sialic acid in the GM3 ganglioside impairs tumor development in mouse lymphocytic leukemia cells | |
EP0381310A1 (en) | Monoclonal antibodies directed to tumor-associated gangliosides and fucogangliosides and method for production thereof | |
EP0972782B1 (en) | Monoclonal antibody which recognizes the oligosaccharide n-glycolylated-galactose-glucose sialic acid in malignant tumors, and composition containing it | |
Robbe-Masselot et al. | Expression of a core 3 disialyl-Lex hexasaccharide in human colorectal cancers: a potential marker of malignant transformation in colon | |
AU659808B2 (en) | Inhibition of metastasis potential and invasiveness by oligosaccharides or oligosaccharide antigens or antibodies | |
Jin et al. | Pathophysiological roles and applications of glycosphingolipids in the diagnosis and treatment of cancer diseases | |
Watarai et al. | Production of monoclonal antibodies directed to Hanganutziu-Deicher active gangliosides, N-glycolylneuraminic acid-containing gangliosides | |
WO1993017033A1 (en) | Inhibition of cell adhesion by chemically-defined oligosaccharides, their derivatives, mimetics, and antibodies directed thereto | |
US6083929A (en) | Extended type 1 chain glycosphingolipids as tumor-associated antigens | |
Ørntoft et al. | Blood group ABH‐related antigens in normal and malignant bladder urothelium: Possible structural basis for the deletion of type‐2 chain ABH antigens in invasive carcinomas | |
TW202333798A (en) | Antibody-drug conjugate targeting claudin 18.2 | |
EP0521692A2 (en) | Inhibition of tumor cell metastasis potential and invasiveness by chemically-defined oligosaccharides, their derivatives, mimetics and antibodies directed to them | |
Satoh et al. | Glycolipid expression in prostatic tissue and analysis of the antigen recognized by antiprostatic monoclonal antibody APG1 | |
KR100666238B1 (en) | Extended type 1 chain glycosphingolipids as tumor-associated antigens | |
US5418129A (en) | Blood treatment method | |
Hollander | Complex N-Glycosylation in Anti-Tumor Immunity | |
WO1992019634A1 (en) | Extended type 1 chain glycosphingolipids as tumor-associated antigens | |
AU2000260652A1 (en) | Extended type 1 chain glycosphingolipids as tumor-associated antigens | |
Smith | Discovery and Characterization of Glyco-Immune Checkpoints | |
JP3006943B2 (en) | Anti-glycolipid glycan monoclonal antibody | |
Stroud et al. | Human Tumor-Associated Lea-Lex Hybrid Carbohydrate Antigen IV3 (Gal. beta. 1. fwdarw. 3 [Fuc. alpha. 1. fwdarw. 4] GlcNAc) III3FucnLc4 Defined by Monoclonal Antibody 43-9F: Enzymic Synthesis, Structural Characterization, and Comparative Reactivity with Various Antibodies |