JPS6256137B2 - - Google Patents
Info
- Publication number
- JPS6256137B2 JPS6256137B2 JP54142153A JP14215379A JPS6256137B2 JP S6256137 B2 JPS6256137 B2 JP S6256137B2 JP 54142153 A JP54142153 A JP 54142153A JP 14215379 A JP14215379 A JP 14215379A JP S6256137 B2 JPS6256137 B2 JP S6256137B2
- Authority
- JP
- Japan
- Prior art keywords
- tumor
- antibody
- antitumor
- antibodies
- solution
- 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.)
- Expired
Links
- 239000000126 substance Substances 0.000 claims description 47
- 206010028980 Neoplasm Diseases 0.000 claims description 38
- 239000002246 antineoplastic agent Substances 0.000 claims description 36
- 230000000259 anti-tumor effect Effects 0.000 claims description 32
- 239000000427 antigen Substances 0.000 claims description 29
- 102000036639 antigens Human genes 0.000 claims description 29
- 108091007433 antigens Proteins 0.000 claims description 29
- 230000000340 anti-metabolite Effects 0.000 claims description 27
- 229940100197 antimetabolite Drugs 0.000 claims description 27
- 239000002256 antimetabolite Substances 0.000 claims description 27
- 238000001042 affinity chromatography Methods 0.000 claims description 25
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 claims description 13
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 12
- 238000000746 purification Methods 0.000 claims description 12
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 claims description 11
- 125000003277 amino group Chemical group 0.000 claims description 11
- 229960000684 cytarabine Drugs 0.000 claims description 11
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 claims description 10
- 108060003951 Immunoglobulin Proteins 0.000 claims description 9
- 150000001875 compounds Chemical class 0.000 claims description 9
- 229960002949 fluorouracil Drugs 0.000 claims description 9
- 102000018358 immunoglobulin Human genes 0.000 claims description 9
- LPXQRXLUHJKZIE-UHFFFAOYSA-N 8-azaguanine Chemical compound NC1=NC(O)=C2NN=NC2=N1 LPXQRXLUHJKZIE-UHFFFAOYSA-N 0.000 claims description 7
- 239000007924 injection Substances 0.000 claims description 7
- 238000002347 injection Methods 0.000 claims description 7
- 229960005508 8-azaguanine Drugs 0.000 claims description 6
- 239000004480 active ingredient Substances 0.000 claims description 6
- 229960004155 aminopterin sodium Drugs 0.000 claims description 6
- USOUFAHHWNKDJC-LTCKWSDVSA-L disodium;(2s)-2-[[4-[(2,4-diaminopteridin-6-yl)methylamino]benzoyl]amino]pentanedioate Chemical compound [Na+].[Na+].C1=NC2=NC(N)=NC(N)=C2N=C1CNC1=CC=C(C(=O)N[C@@H](CCC([O-])=O)C([O-])=O)C=C1 USOUFAHHWNKDJC-LTCKWSDVSA-L 0.000 claims description 6
- 208000032839 leukemia Diseases 0.000 claims description 6
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 claims description 3
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 claims description 3
- 201000009030 Carcinoma Diseases 0.000 claims description 3
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 claims description 3
- 201000006491 bone marrow cancer Diseases 0.000 claims description 3
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 2
- 208000009916 Yoshida Sarcoma Diseases 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 238000004587 chromatography analysis Methods 0.000 claims description 2
- 201000005202 lung cancer Diseases 0.000 claims description 2
- 208000020816 lung neoplasm Diseases 0.000 claims description 2
- 208000006268 Sarcoma 180 Diseases 0.000 claims 1
- 239000002260 anti-inflammatory agent Substances 0.000 claims 1
- 229940124599 anti-inflammatory drug Drugs 0.000 claims 1
- 229960003058 methotrexate sodium Drugs 0.000 claims 1
- 239000000243 solution Substances 0.000 description 40
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 25
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 22
- 210000004027 cell Anatomy 0.000 description 22
- 238000006243 chemical reaction Methods 0.000 description 21
- 239000007864 aqueous solution Substances 0.000 description 19
- 238000000034 method Methods 0.000 description 19
- 210000004881 tumor cell Anatomy 0.000 description 19
- 241000283973 Oryctolagus cuniculus Species 0.000 description 16
- 239000012153 distilled water Substances 0.000 description 15
- 239000006228 supernatant Substances 0.000 description 15
- 239000002953 phosphate buffered saline Substances 0.000 description 13
- 238000011765 DBA/2 mouse Methods 0.000 description 11
- 235000017557 sodium bicarbonate Nutrition 0.000 description 11
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 11
- 238000009739 binding Methods 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 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 239000011521 glass Substances 0.000 description 9
- 229960000485 methotrexate Drugs 0.000 description 9
- 241000699670 Mus sp. Species 0.000 description 8
- 239000007795 chemical reaction product Substances 0.000 description 8
- 239000000385 dialysis solution Substances 0.000 description 8
- 239000002244 precipitate Substances 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 7
- 230000015572 biosynthetic process Effects 0.000 description 7
- 238000000502 dialysis Methods 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 238000003786 synthesis reaction Methods 0.000 description 7
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 6
- 208000003455 anaphylaxis Diseases 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 239000007758 minimum essential medium Substances 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 206010002199 Anaphylactic shock Diseases 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 5
- 229940041181 antineoplastic drug Drugs 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 231100000263 cytotoxicity test Toxicity 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 206010003445 Ascites Diseases 0.000 description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 229920002684 Sepharose Polymers 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 235000019441 ethanol Nutrition 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 239000007974 sodium acetate buffer Substances 0.000 description 4
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 210000004988 splenocyte Anatomy 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 230000001988 toxicity Effects 0.000 description 4
- 231100000419 toxicity Toxicity 0.000 description 4
- 230000004614 tumor growth Effects 0.000 description 4
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 3
- IVLXQGJVBGMLRR-UHFFFAOYSA-N 2-aminoacetic acid;hydron;chloride Chemical compound Cl.NCC(O)=O IVLXQGJVBGMLRR-UHFFFAOYSA-N 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 239000004375 Dextrin Substances 0.000 description 3
- 229920001353 Dextrin Polymers 0.000 description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- 229920005654 Sephadex Polymers 0.000 description 3
- 239000012507 Sephadex⢠Substances 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000000862 absorption spectrum Methods 0.000 description 3
- 239000011543 agarose gel Substances 0.000 description 3
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 239000003054 catalyst Substances 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 235000019425 dextrin Nutrition 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 229960004857 mitomycin Drugs 0.000 description 3
- 239000008057 potassium phosphate buffer Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000013076 target substance Substances 0.000 description 3
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 2
- 206010000830 Acute leukaemia Diseases 0.000 description 2
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 241000700199 Cavia porcellus Species 0.000 description 2
- 206010010904 Convulsion Diseases 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 206010025323 Lymphomas Diseases 0.000 description 2
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 description 2
- 206010039491 Sarcoma Diseases 0.000 description 2
- RJURFGZVJUQBHK-UHFFFAOYSA-N actinomycin D Natural products CC1OC(=O)C(C(C)C)N(C)C(=O)CN(C)C(=O)C2CCCN2C(=O)C(C(C)C)NC(=O)C1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)NC4C(=O)NC(C(N5CCCC5C(=O)N(C)CC(=O)N(C)C(C(C)C)C(=O)OC4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-UHFFFAOYSA-N 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 239000005557 antagonist Substances 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 150000001718 carbodiimides Chemical class 0.000 description 2
- FOCAUTSVDIKZOP-UHFFFAOYSA-N chloroacetic acid Chemical compound OC(=O)CCl FOCAUTSVDIKZOP-UHFFFAOYSA-N 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 230000036461 convulsion Effects 0.000 description 2
- 230000001472 cytotoxic effect Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 238000000921 elemental analysis Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 230000003211 malignant effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 201000001441 melanoma Diseases 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 239000003021 water soluble solvent Substances 0.000 description 2
- ILFPCMXTASDZKM-YFKPBYRVSA-N (1s)-2-methylidene-3-oxocyclopentane-1-carboxylic acid Chemical compound OC(=O)[C@H]1CCC(=O)C1=C ILFPCMXTASDZKM-YFKPBYRVSA-N 0.000 description 1
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 1
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 description 1
- 201000004384 Alopecia Diseases 0.000 description 1
- 206010002198 Anaphylactic reaction Diseases 0.000 description 1
- 108010006654 Bleomycin Proteins 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 1
- 108010092160 Dactinomycin Proteins 0.000 description 1
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 1
- MWWSFMDVAYGXBV-RUELKSSGSA-N Doxorubicin hydrochloride Chemical compound Cl.O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 MWWSFMDVAYGXBV-RUELKSSGSA-N 0.000 description 1
- 239000006145 Eagle's minimal essential medium Substances 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 208000018522 Gastrointestinal disease Diseases 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
- 208000008342 Leukemia P388 Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 208000015634 Rectal Neoplasms Diseases 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- RJURFGZVJUQBHK-IIXSONLDSA-N actinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-IIXSONLDSA-N 0.000 description 1
- 230000007059 acute toxicity Effects 0.000 description 1
- 231100000403 acute toxicity Toxicity 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 230000036783 anaphylactic response Effects 0.000 description 1
- 238000011091 antibody purification Methods 0.000 description 1
- 239000003125 aqueous solvent Substances 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 229960001561 bleomycin Drugs 0.000 description 1
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 210000001715 carotid artery Anatomy 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 230000002301 combined effect Effects 0.000 description 1
- 239000000306 component Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 229960000640 dactinomycin Drugs 0.000 description 1
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 description 1
- 229960000975 daunorubicin Drugs 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 229960002918 doxorubicin hydrochloride Drugs 0.000 description 1
- 239000013583 drug formulation Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 239000007973 glycine-HCl buffer Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 208000024963 hair loss Diseases 0.000 description 1
- 230000003676 hair loss Effects 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 229940072221 immunoglobulins Drugs 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 201000002364 leukopenia Diseases 0.000 description 1
- 231100001022 leukopenia Toxicity 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- -1 methotoxate Chemical compound 0.000 description 1
- IZXGZAJMDLJLMF-UHFFFAOYSA-N methylaminomethanol Chemical compound CNCO IZXGZAJMDLJLMF-UHFFFAOYSA-N 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- SJXULIRYGQTEJK-UHFFFAOYSA-N n'-(2-morpholin-4-ylethyl)methanediimine Chemical compound N=C=NCCN1CCOCC1 SJXULIRYGQTEJK-UHFFFAOYSA-N 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 210000003200 peritoneal cavity Anatomy 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000000722 protumoral effect Effects 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 206010038038 rectal cancer Diseases 0.000 description 1
- 201000001275 rectum cancer Diseases 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 210000002235 sarcomere Anatomy 0.000 description 1
- ILFPCMXTASDZKM-UHFFFAOYSA-N sarkomycin Natural products OC(=O)C1CCC(=O)C1=C ILFPCMXTASDZKM-UHFFFAOYSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 229940083542 sodium Drugs 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000012089 stop solution Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000003799 water insoluble solvent Substances 0.000 description 1
Landscapes
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Description
æ¬çºæã¯æ°èŠæè
«çå€ã«é¢ããæŽã«è©³ããã¯ã
è «çæåã«å¯Ÿããæäœã«ãæè «çæ§ä»£è¬æ®æå€ã
ã¢ããçµåãããŠãªãç©è³ªãæå¹æåãšããŠå«æ
ããæè «çå€ã«é¢ããã æ¬çºæè ã¯ãããã«ãã€ããã€ã·ã³ïŒ£ãå¡©é žã
ããœã«ãã·ã³ããã¬ãªãã€ã·ã³ãããŠãã«ãã·
ã³ãã¢ã¯ãããã€ã·ã³ïŒ€ããã³ã¶ã«ã³ãã€ã·ã³ã®
ããšãæçç©è³ªç³»ã®æè «çå€ãè «çæäœã«çµåã
ãŠãªãç©è³ªãæå¹æåãšããæè «çå€ãçºæãã
ïŒç¹é¡æ53â161388åç §ïŒã ããããäžèšæçç©è³ªç³»ã®æè «çå€ã¯äž»ãšããŠ
埮çç©ã«ããçç£ãããç©è³ªã§ãã€ãŠæ§é ãè€é
ã§ããããããããè «çæäœãšçµåãããŠæè «ç
æ§ç©è³ªã補é ããããã§æé©ã®ãã®ãšã¯èšããª
ãã æ¬çºæè ã¯æŽã«ç 究ããçµæãã·ã¿ã©ãã³ãã¡
ãœããã»ãŒããã¢ãããããªã³ãããªãŠã ãïŒâ
ã¢ã¶ã°ã¢ãã³ãïŒâãã«ãªããŠã©ã·ã«ã®ããšã代
è¬æ®æå€ç³»ã®ç©è³ªãè «çæäœãšçµåãããŠæè «ç
æ§ç©è³ªã補é ããã«é©ããŠããäºãèŠãã ãæ¬çº
æããªãã«è³ã€ãã æ¬çºæã¯ã现èæ¯æ§ã®é«ãäžæ²æè «çæ§ä»£è¬æ®
æç©è³ªã極ããŠç©åãªæ¡ä»¶äžã§è «çæåã«å¯Ÿãã
æäœãšã¢ããçµåïŒâNHCOâïŒã«ãã€ãŠçµåã
ããæ°èŠãªç©è³ªãæå¹æåãšããæè «çå€ã«ä¿ã
ãã®ã§ãã€ãŠãæè «çå¹æã«ããããªãã现èæ¯
æ§ã¯ãåºçºç©è³ªã®ïŒã€ã§ããäžèšæè «çæ§ä»£è¬æ®
æå€ã«ããã¹ãŠæ Œæ®µã«äœãæè «çå€ãæäŸããã
ãšãç®çãšããã ããã«ãæ¬çºæã¯è «çæåã«å¯Ÿããæäœèªèº«ã®
粟補æ段ããã³ããã粟補æäœãã¢ããåºåã¯ã«
ã«ããã·ã«åºãæããäžèšã®æè «çæ§ä»£è¬æ®æç©
質åã³ã¢ããåºåã¯ãã«ã«ããã·ã«åºãå°å ¥ãã
äžèšã®æè «çæ§ä»£è¬æ®æç©è³ªã«ã¢ããçµåïŒâ
NHCOâïŒã«ãã€ãŠçµåãããããšãããªãäžèš
ã®ç©è³ªãæå¹æåãšããæè «çå€ã補é ããæ¹æ³
ãæäŸããããšãç®çãšããã è¿å¹Žãçš®ã ã®æè «çå€ãåºã䜿çšãããŠããã
æãçšåºŠã®å¹æããããŠããããããã®æè «çå€
ãšããŠã·ã¿ã©ãã³ãã¡ãœãã¬ãã»ãŒããã¢ããã
ããªã³ãããªãŠã ãïŒâã¢ã¶ã°ã¢ãã³ãïŒâãã«
ãªããŠã©ã·ã«ã®ããšã代è¬æ®æå€ã䜿çšãããŠã
ããããããã®ç©è³ªã¯ããèªäœäœããé«ã现èæ¯
æ§ãæããŠããŠãæäžããæã«çœè¡çæžå°ãè±
æ¯ãèè žé害çã®å¯äœçšãåããããšãç¥ãããŠ
ããããã®çºã«ãããè¬å€ã®äœ¿çšã«é床ã®ããã®
ãå®æ ã§ããã ãŸããåŸæ¥ããããçš®ã®è «çæåã«å¯Ÿããæäœ
ã補é ãŸãã¯åé¢ããŠããããã®è «çã®æ²»çã«çš
ããè©Šã¿ããªãããŠããããæãŸããæè «çå¹æ
ã¯åŸãããŠããªããããã«ãæè¿è «çæäœã«æè «
çå€ãååŠçã«çµåãããŠåŸãããæ°èŠãªç©è³ªã«
ããæè «çå¹æãæåŸ ããããšãææ¡ãããŠãã
ããäžèšç©è³ªãåŸãããã®ååŠåå¿ã®æ¡ä»¶ãéé ·
ãããããã«ååãªæé·ã¯åŸãããŠããªãããŸ
ãããããã®å®éšã§çšããããæäœã¯å ç«ã°ãã
ãªã³ç»åãŸã§ã®ç²Ÿè£œããè¡ãªãããŠããªãã®ã§ã
äžè¬ã®å ç«ã°ãããªã³ãå«æããŠããŠãçã®æå³
ã§ã¯è «çæäœãšã¯èªãé£ãããããŠæäœãäžè¬ã®
å ç«ã°ãããªã³ã§ããæ ã«äžèšååç©ãæäžãã
ãçäœã«æ£åžžçµç¹ã®é害ãå šèº«çæ£ãŸãã¯ç¡¬çŽãª
ã©ã®ã¢ããã€ã©ãã·ãŒã·ãšãã¯ã®çããäºãå€
ãã æ¬çºæã¯ãåç©ã«æ¥çš®ããè «çã®æåã«å¯Ÿãã
æäœãå«ãæè¡æž ãæ¡åãããã®äžã®å ç«ã°ãã
ãªã³ç»åãååãããããæŽã«ç¹ç°ãªæ段ã§ç²Ÿè£œ
ããããšã«ãã€ãŠãããçŽç²ãªè «çæäœãå ã¥ã
åé¢ãããããæè «çæ§ä»£è¬æ®æå€ãšçµåããã
ããšã«ãããç©åãªåå¿æ¡ä»¶äžã§è£œé ã§ãããã€
äžèšã®ããšãã¢ããã€ã©ãã·ãŒã·ãšãã¯ãçããª
ãæè «çå€ãæäŸãåŸããã®ã§ããã 以äžæ¬çºæã詳ãã説æããã æ¬çºæã®æå¹æåã§ããç©è³ªã¯ãæäœããã³æ
è «çæ§ä»£è¬æ®æå€ã®äž¡è ã«å æ¥ååšããŠããã¢ã
ãåºåã¯ã«ã«ããã·ã«åºãç©åãªåå¿æ¡ä»¶äžã«å
å¿ããããããšã«ããããã®äž¡è ãã¢ããçµåã
ããŠåŸãããããã®éãå¿ èŠã«ããäžèšä»£è¬æ®æ
å€ã«ããã«ã¢ããåºåã¯ã«ã«ããã·ã«åºãå°å ¥ã
ããã®ãæäœãšåå¿ãããããšã«ããäž¡è ãã¢ã
ãçµåãããã æ¬ç©è³ªã®è£œé ã«çšããæè «çæ§ä»£è¬æ®æå€ãšã
ãŠã¯ã·ã¿ã©ãã³ãã¡ãœãã¬ãã»ãŒããã¢ãããã
ãªã³ãããªãŠã ãïŒâã¢ã¶ã°ã¢ãã³ãªãã³ã«ïŒâ
ãã«ãªããŠã©ã·ã«ã奜ãŸããã äžæ²ã®æè «çæ§ä»£è¬æ®æå€ã®å称ã¯äžè¬åã§ã
ã€ãŠãããã®æ§é ã¯æ¬¡ã®ãšããã§ããã ã·ã¿ã©ãã³ïŒ ã¡ãœãã¬ãã»ãŒãïŒ ã¢ãããããªã³ãããªãŠã ïŒ ïŒâã¢ã¶ã°ã¢ãã³ïŒ ïŒâãã«ãªãã©ãŠã·ã«ïŒ ãŸããæ¬ç©è³ªã®åºçºç©è³ªã®ïŒã€ã§ããæäœãšã
ãŠã¯æ²»çãç®çãšããè «çã®æäœãçšããããã è «çãšããŠã¯ã¶ã«ã³ãŒããŒ180ãäœè€èºçãå
ç°èè «ããšãŒãªããçãâ1210çœè¡ç ãâ
388çœè¡ç ãæ¥æ§çœè¡ç ãæªæ§ãªã³ãè «ãçè «ã
èè «ãæªæ§ç¹æ¯äžç®è «ãæ¥æ§éªšé«æ§çœè¡ç ãã¡ã©
ããŒããæ¥æ§ãªã³ãæ§çœè¡ç ã骚é«çããªã©ã®å
çš®ã®è «çããããããã æ¬çºæã§çšããæäœã®è£œé ã¯æ¥æ¬å ç«åŠäŒç·äŒ
èšé²ç¬¬ïŒå·»198é ïŒ1976幎ïŒèšé²ã®æ¹æ³ã«ããå
ã¯DauphinãM.J.ãã®æ¹æ³ãJ.Immunal.113948
ïŒ1974åç §ïŒãã«æºããŠè¡ãªããåè ã¯ããã€ã³ã
ã®ã³ã³ããªãŒãã¢ãžãŠãã³ãïŒComplete
AjuvantïŒãçšããå ãè «ç现èãåç©ã«ç®äžæ³š
å°ããããšã«ãã€ãŠããããå ç«ãæŽã«ç¶ããŠè «
ç现èãéèå 泚å°ããããšã«ããè¿œå å ç«ããŠ
ããããæäœãåŸãæ¹æ³ã§ãããåŸè ã¯è «çæå
ãåç©ã®è ¹è å ã«ïŒãïŒåå埩æäžããŠåç©ãå
ç«ããŠãããããæäœãåŸãæ¹æ³ã§ããã æ¬çºæã§çšããæäœã¯åçš®æäœãç°çš®æäœã®ã
ããã§ãããããåçš®æäœã奜ãŸããã ãã®ããã«ããŠåŸãæäœã¯ãã¢ãã€ããã€ã¯ã
ããã°ã©ãã€ãŒã«ãããã«ã©ã ã®å å¡«å€ã«æ²»çã®
ç®çã®è «çæåãå ã¥ããã ã·ã¢ã³ãçšããŠçµå
ããããŠçœ®ããŠããããã«ã©ã ã«å å¡«ããã«ã©ã
ã«å ç«ã°ãããªã³ç»åãŸã§ç²Ÿè£œããæäœã®æº¶æ¶²ã
æµå ¥ãããããããŠã«ã©ã äžã§æåãšæäœãšãçµ
åãããåŸã«ãç¹æ®ãªæº¶å€ãã«ã©ã ã«æµå ¥ããŠæ
åã»æäœçµåã解ãããŠæäœã®ã¿ã溶åºãããæŽ
ã«ãããéæããŠç²Ÿè£œæäœæ°Žæº¶æ¶²ãåŸããå³ã¡ã
ãããŠåŸãããæäœã¯åŸæ¥ã®å ç«ã°ãããªã³ç»å
ãããæŽã«çŽåºŠã®é«ãæè «çå ç«ã°ãããªã³ã§ã
ãã äžè¬ã«æäœã®ç²Ÿè£œã«ã¯ãç¡«å®å¡©æãšDEAEã»ã«
ããŒã¹ã«ã©ã ã«ããã€ãªã³äº€æã¯ãããã°ã©ãã€
ãŒã䜿ã€ãŠãæè¡æž ããåç»ãåŸãæ¹æ³ããã°ã
ã°çšãããããæ¬çºæã§ã¯ãããã«å ããŠãã¢ã
ã€ããã€ã¯ãããã°ã©ãã€ãŒãçšããŠè «ç现èã«
察ããç¹ç°çæäœã®ã¿ãéžæçã«åŸãçºã®ç¹ç°ç
粟補æäœãè¡ãªã€ãã ã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒã¯ãé µçŽ ãšåº
質ãæäœãšæåãªã©ã®ãããªçäœç©è³ªçžäºéã«å
ãç¹ç°ç芪ååãå©çšããäžæ¹ã®çäœç©è³ªã䜿ã€
ãŠä»æ¹ãéžæçã«åé¢ãããšããåçã«ããšã¥ã
ãã®ã§ããã æ¬çºæã§çšããããã¢ãã€ããã€ã¯ãããã°ã©
ãã€ãŒã«ã¯ã(1)è «ç现èããæœåºããæåãã»ã
ã¢ããŒã¹ïŒSepharoseRïŒã®ããšãæ äœã«ïŒãã
ã ã·ã¢ã³ãçšããŠïŒå ±æçµåããããããã«ã©ã
ã«å å¡«ããæäœæº¶æ¶²ãéããŠãæäœãæåã«çµå
ãããæŽã«ååéã®æº¶åªãæµããŠçµåããªãã€ã
æäœãæŽãå»ã€ãåŸãPHã®ããäœã緩衡溶液ãæµ
ãå ¥ããŠãæäœã»æåã®çµåã解ããŠãåé¢ãã
æäœã溶åºãããæ¹æ³ã(2)ã«ã©ã ãçšãããæå
ãçµåãããæ äœãšæäœæº¶æ¶²ãæ··åããŠæäœãæ
åã«çµåãããæ äœç²åãæŽæµããŠçµåããªãã€
ãæäœãé€ããåŸãæäœã溶é¢ãããæ¹æ³ããã
ã³(3)äžèšã§æåãçµåãããæ äœã®ä»£ãã«ãè «ç
现èèªèº«ãçšããæ¹æ³ãå«ããæ¬çºæã®å®æœäŸã§
ã¯äžèš(1)ãš(3)ã®æ¹æ³ãæ¡çšããã åŸã€ãŠãã¢ãã€ããã€ã¯ãããã°ã©ãã€ã«ãã€
ãŠç²Ÿè£œãããæäœã¯åŸæ¥ã®å ç«ã°ãããªã³ç»åã
ããé¥ã«çŽåºŠã®é«ãæè «çå ç«ã°ãããªã³å³ã¡æ
è «çæäœã§ããã ãã®ããã«ããŠåŸãããæäœãæè «çæ§ä»£è¬æ®
æå€ã«çµåãããã«ã¯ãã¢ããåºåã¯ã«ã«ããã·
ã«åºãæããäžèšä»£è¬æ®æå€ãããã¯ã¢ããåºå
ã¯ã¢ããåºãå°å ¥ããäžèšä»£è¬æ®æå€ãšã氎溶æ§
溶åªäžã«æº¶è§£ããããããã«ã«ã«ããžã€ããã觊
åªãšããŠå ããŠïŒã50âã奜ãŸããã¯10ã40âã§
10åãïŒæéã奜ãŸããã¯30åãïŒæéåå¿ãã
ãŠé ¢é žâé ¢é žãããªãŠã ç·©è¡æ¶²ãªã©ã®æ·»å ã§åå¿
ãåæ¢ãããã 次ã«ãã®åå¿æ¶²äžã®éå°ãªä»£è¬æ®æå€ã觊åªã
ãã³äžèšåå¿åæ¢æ¶²ã®æåãªãã³ã«å¡©é¡ãé€ãçº
ã«éæãã²ã«éãªãã³ã«éå€éã®äœããã®æ
äœãè¡ãªãããåã¯ãããã®æäœãçµã¿åããæ
äœãè¡ãªãã äžèšåå¿ã§è§ŠåªãšããŠçšããã«ã«ããžã€ãã
ã¯ãïŒâãšãã«âïŒâïŒïŒâãžã¡ãã«ã¢ãããã
ãã«ïŒâã«ã«ããžã€ãããïŒâã·ã¯ãããã·ã«â
ïŒâïŒïŒâã¢ã«ããªããšãã«ïŒã«ã«ããžã€ããå
ã¯ãžã·ã¯ãããã·ã«ã«ã«ããžã€ãããå«ãã æè «çæ§ä»£è¬æ®æå€ã«å¿ èŠã«ããã¢ããåºåã¯
ã«ã«ããã·ã«åºãå°å ¥ããã«ã¯ã該æ®æå€ãã®ãŸ
ãŸã«ãåã¯NaããAgã®ããšãå¡©ã®åœ¢äœã«ãã
ãã®ã«ãäžè¬åŒïŒžïŒCH2ïŒoCOOHïŒåŒäžïŒžã¯Clå
ã¯Brã瀺ããïœã¯ïŒä¹è³ïŒã®æŽæ°ã瀺ãïŒã®å
åç©ãæãã¯äžè¬åŒHClã»NH2ïŒCH2ïŒoCOXïŒåŒ
äžïŒžã¯Clåã¯Brã瀺ããïœã¯ïŒä¹è³ïŒã®æŽæ°ã
瀺ãïŒã®ååç©ãæ°Žæ§æº¶åªãäŸãã°ã¡ã¿ããŒã«ã
æ°Žããšã¿ããŒã«ããžã¡ãã«ã¹ã«ããªãã·ãããžãª
ããµã³äžã§ïŒã50âã奜ãŸããã¯10ã40âã§10å
ä¹è³72æéåå¿ããããšãããåå¿çæç©ã¯æ°Žã
ã¢ã«ã³ãŒã«ãã¯ãããã«ã ããžãªããµã³ã®ããšã
溶åªãçšããŠåçµæ¶ããããšã«ããæè «çæ§ä»£è¬
æ®æå€èªå°äœãåŸãããã æ¬ç©è³ªã¯æäœïŒååã«å¯ŸããŠæè «çæ§ä»£è¬æ®æ
å€ã®ïŒã10ååãçµåãããã®ã§ããã æ¬ç©è³ªã®åºä¹³åç©ã«å¯Ÿããæ¥æ§æ¯æ§ã¯ããŠã¹ã
çšã300mgïŒKgã®éèæäžã§èª¿ã¹ããïŒé±éã®èŠ³
å¯ã§æ»äº¡ãèªããããªãã€ãã ãããã€ãŠãæ¬ç©è³ªã¯æ¯æ§ãäœãåçš®ã®äººçã«
察ããŠæå¹ã§ãããäŸãã°ãæ¥æ§çœè¡ç ãæªæ§ãª
ã³ãè «ãçè «ãèè «ãæªæ§ç¹æ¯äžç®è «ãæ¥æ§éªšé«
æ§çœè¡ç ãã¡ã©ããŒããæ¥æ§ãªã³ãæ§çœè¡ç ã骚
é«ççã«æå¹ã§ããã æ¬ç©è³ªãæè «çå€ãšããŠçšããå Žåã®è£œå€åæ¹
æ³ãããã³æäžã®æ¹æ³ãšããŠã¯æè «çå€ã«é¢ãã
å ¬ç¥ã®æ¹æ³ãé©çšãåŸããæäžæ¹æ³ãšããŠã¯çµ
å£ã泚å°ãŸãã¯çŽè žæäžãããããæäžåœ¢æ ãšã
ãŠã¯ç²æ«ãé¡ç²ãé å€ãŸãã¯æ³šå°å€ã座è¬çã®ã
ããã§ãã€ãŠãããã ç¹ã«æ³šå°ã«ããæäžã奜ãŸããã 泚å°è¬ã®è£œå€ã«ã¯çççé£å¡©æ°Žãæ» èæ°Žããªã³
ã²ã«æ¶²çã®æ°Žæº¶æ§æº¶å€ãé氎溶æ§æº¶å€ãç匵å
å€ãç¡çåå€ãå®å®å€ãé²è å€ãæžæ¿åå€ãç·©è¡¡
å€ãä¹³åå€çãä»»æã«çšãåŸãã ãã®äžäŸã瀺ããšãæ¬ç©è³ª10mgãšãã³ãããŒã«
50mgãèžæºæ°Žã«æº¶è§£ããŠ10mlãšããŠåžžæ³ã§é€èã
ãåŸïŒmlå®ã泚å°çšå°ç¶ã«å泚ããåã¯ãã®ãŸãŸ
åçµä¹Ÿç¥ããŠæ³šå°å€ãšããããããŠæ¬å€ã¯äœ¿çšã«
éããçççé£å¡©æ°Žã§çšéããŠæ³šå°æ¶²ãšããã æ¬ç©è³ªã¯è£œå€åäžäžè¬ã«0.01ã90ïŒ å¥œãŸããã¯
0.1ã60ïŒ å«æããããšãã§ããã æ¬ç©è³ªã®æäžéã¯äž»ãšããŠçç¶ã«å·Šå³ãããã
æ人ïŒäººïŒæ¥åœã0.1ãïŒïœã奜ãŸããã¯ïŒãïŒ
ïœã§ããã ãªããæ¬çºæã«ãããšãæäœã®åè «çæ§ãšæè «
çå€ãªãã³ã«æè «çå€ã®æè «çæ§ã¯å€±ãããããš
ãªãäžèšååç©ã«ãã®ãŸãŸä¿ãããŠããã®ã§ãæ¬
ç©è³ªã¯æäžããããšå¹çããç®çãšããè «çéšäœ
ã«å°éããæè «çå¹æãçºæ®ãããåŸã€ãŠæ¬ç©è³ª
ããã®æåãšããŠå«æããæè «çå€ã®ééãåºæº
ãšããŠèãããªãã°ãåäžã®æè «çå€ãã®ãã®ã®
æäžéã®1/10ã1/20ã«çžåœããééã®æè «çå€ã
æåãšããŠå«æããæ¬ç©è³ªã®æäžã«ãã€ãŠåçšåºŠ
ã®è «çå¢æ®æå¶ãåŸããããã€æåãšããŠå«æã
ãæè «çå€ã«ããå¯äœçšã¯ãåäžã®æè «çå€ãã®
ãã®ã®æäžã®å Žåã®1/10ã2/10ã«ãããªããšæåŸ
ããããããã¯æ¬ç©è³ªã®æåããããã®å¥œãŸãã
æ§è³ªã®è€åå¹æãšäºãããšãã§ããã 以äžæ¬çºæãå®æœäŸã«ãã€ãŠæŽã«è©³çŽ°ã«èª¬æã
ãã å®æœäŸ ïŒ ïŒâïŒïŒã¢ãã€ããã€ã¯ãããã°ã©ãã€(3)ãå©çš
ããæäœã®èª¿è£œãšç²Ÿè£œ DBAïŒïŒããŠã¹ãçšããŠç¶ä»£å¹é€ããè ¹
æ°Žåâ388è «ç现èãçççé£å¡©æ°Žã«æžæ¿
ããããã€ããã€ã·ã³ïŒ£ïŒ50ÎŒïœïŒmlïŒãå
ããŠ30åé37âã§åŠçããã®ã¡é å¿åé¢ã«ã
ã€ãŠäžæž ãé€ãã现èã0.85ïŒ ã®ççé£å¡©æ°Ž
ã§ïŒåæŽæ»ãããããããŠå¢æ®èœã倱ã€ã
â388è «ç现èã«ããã€ã³ãïŒFreundïŒã®ã³
ã³ããªãŒãã¢ãžãŠãã³ãïŒComplete
AjuvantïŒïŒFCAãšç¥ãïŒãæ··åããäœé2.9
Kgã®ãŠãµã®è¶³è¹ ã®ç®äžã«ïŒå¹åœã现è108å
ã®å²åã§æ³šå°ããïŒé±éåŸã«åæ§ãªæ¹æ³ã§æ³š
å°ããŠãŠãµã®ãå ç«ããæŽã«ïŒé±éåŸåäžã®
现è108åããã®ãŠãµã®ã®éèã«æ³šå°ããã
ïŒé±éçµéåŸã«é åèã«ã«ããŠãŒã¬ãæ¿å ¥ã
ãŠå šè¡ãæ¡åããããããæè¡æž ãåé¢ããŠ
äžèšã®ç²Ÿè£œãè¡ã€ããå³ã¡ã100mlã®æè¡æž
ã«ç¡«é žã¢ã³ã¢ããŠã ã飜åéã®20ã30ïŒ æ·»å
ããŠçããå¡©æç»åãæ¡åããŠãããã20ml
ã®æ°Žã«å溶解ãããã®æº¶æ¶²ãPH7.0ã®10ïœïŒ
çé žå¡©ç·©è¡é£å¡©æ°Žæº¶æ¶²ïŒPBSãšç¥ãïŒã«å¯Ÿã
ãŠïŒâã72æééæããŠè±å¡©ããïŒãã®éã
24æéæ¯ã«éæå€æ¶²ã®äº€æãè¡ã€ãïŒããã
ã«ãé£å¡©æ°Žã§ïŒåæŽæµããæ£åžžãªDBAïŒïŒ
ããŠã¹ã®è¡çãçéæ··åããïŒâã§30åæŸçœ®
ããŠåžåãè¡ããããåŸãé å¿åé¢ããŠäžæž
ãåŸãããã®åžåæäœãæŽã«ïŒåè¡ã€ãŠãå š
éšã§ïŒåã®åžåæäœãå ããããããŠåŸãã
ãæäœã粟補åæäœãšäºãïŒé垞ïœïŒ§ïŒã
次ãã§äžèšã®æ¹æ³ã§æŽã«ç²Ÿè£œãè¡ã€ããå³ã¡
äžèšåžåã®æžãã äžæž ã«ãâ388现èãç
éæ··åããïŒâã«30åæŸçœ®ããŠïŒ°â388现è
ã«å¯Ÿããæäœãâ388è «ç现èã«çµåãã
ãåŸãé å¿åé¢ããŠäžæž ãé€ããæ²æŸ±ã«PH
3.0ã®ã°ãªã·ã³âå¡©é žç·©è¡æ¶²ãå ããŠãæäœ
ãéé¢ãããããã®æ··åç©ãé å¿åé¢ããŠã
æäœãå«ãäžæž ãæ¡åãããäžæž ã®PHã
0.1Mèæ§ãœãŒã氎溶液ã§äžæ§è¿ãã«èª¿æŽã
ãåŸãPBSã«å¯ŸããŠïŒâã§24æééæãè¡ã€
ãïŒïŒæéæ¯ã«éæå€æ¶²ã亀æããïŒããã
ãŠè¡ããããã®ããŠãµã®ã®æâ388çœè¡ç
è «çå ç«æäœã®æ°Žæº¶æ¶²ã§ããã ïŒâïŒïŒè «ç现èããã³æ£åžžçŽ°èã«å¯Ÿããé害æ§
è©ŠéšïŒãã®ïŒïŒ äžèšã§åŸããŠãµã®ã®æâ388è «çå ç«æ
äœã«ã€ããŠè£äœïŒã¢ã«ã¢ããã®è¡æž ïŒååšäž
ã®çŽ°èé害詊éšãè¡ã€ããå³ã¡ãäžèšæäœæ°Ž
溶液åã¯ããã10ã100ããã³1000åã«çšé
ãããã®ãšãâ388è «ç现èæµ®é液æãã¯
æ£åžžDBAïŒïŒããŠã¹èŸçŽ°èæµ®é液ïŒäœãã
ã€ãŒã°ã«ïŒEagleïŒã®Minimum Essential
MediumïŒä»¥äžMEMãšç¥ãïŒã溶åªãšããŠ
çšãã现èæ¿åºŠã¯ïŒÃ106åïŒãšããïŒãšã
100ÎŒå®æ··åãã宀枩ã§15åæŸçœ®ããŠæäœ
ãããããã®çŽ°èã«çµåããããããã®åŸã¢
ã«ã¢ããã®è¡æž ãã€ãŒã°ã«ã®MEMã§ïŒåã«
çšéãïŒãããè£äœãšäºãïŒãã®100ÎŒ
ããäžèšã®æ··å液ã«å ãã37âã§30åå¹é€
ããé å¿åé¢ããŠãæ²æŸ±ãã€ãŒã°ã«ã®MEM
ã§ïŒåæŽæ»ããããã«ããªãã³ãã«ãŒæ¶²ãå
ããŠãé¡åŸ®é¡äžã§äžèšããããã®çŽ°èã®æ»æ»
ã®çšåºŠã芳å¯ããã ãã®çµæã¯äžèšã®ç¬¬ïŒè¡šã«ç€ºãããšãã现
èã®æ»æ» çšåºŠïŒçŽ°èé害掻æ§ïŒãïŒãïŒïŒãïŒ
ïŒïŒã®ïŒæ®µéã«åããŠè¡šç€ºããïŒæ»æ» ãªãã¯
âã§ç€ºãïŒãšæè¡æž ãäžèšã®æ¹æ³ã§ç²Ÿè£œãã
åŸã«æäœãåãåºãããã®ã¯ãæè¡æž ã粟補
ããããšãªãæäœãåãåºãããã®ã«ããã¹
ãŠïŒ°â388çœè¡ç 现èã«å¯Ÿããæ¯æ§ã¯ããŸã
倧ããç°ããªãããæ£åžžã®DBAïŒïŒããŠã¹
èŸçŽ°èã«å¯Ÿããæ¯æ§ã¯æ¥µããŠäœããããã殺
ãããšã¯ãªãã€ããå³ã¡æè¡æž ã®ç²Ÿè£œã¯ç®ç
ã«ããªã€ãŠããããšããã瀺ãããã
è «çæåã«å¯Ÿããæäœã«ãæè «çæ§ä»£è¬æ®æå€ã
ã¢ããçµåãããŠãªãç©è³ªãæå¹æåãšããŠå«æ
ããæè «çå€ã«é¢ããã æ¬çºæè ã¯ãããã«ãã€ããã€ã·ã³ïŒ£ãå¡©é žã
ããœã«ãã·ã³ããã¬ãªãã€ã·ã³ãããŠãã«ãã·
ã³ãã¢ã¯ãããã€ã·ã³ïŒ€ããã³ã¶ã«ã³ãã€ã·ã³ã®
ããšãæçç©è³ªç³»ã®æè «çå€ãè «çæäœã«çµåã
ãŠãªãç©è³ªãæå¹æåãšããæè «çå€ãçºæãã
ïŒç¹é¡æ53â161388åç §ïŒã ããããäžèšæçç©è³ªç³»ã®æè «çå€ã¯äž»ãšããŠ
埮çç©ã«ããçç£ãããç©è³ªã§ãã€ãŠæ§é ãè€é
ã§ããããããããè «çæäœãšçµåãããŠæè «ç
æ§ç©è³ªã補é ããããã§æé©ã®ãã®ãšã¯èšããª
ãã æ¬çºæè ã¯æŽã«ç 究ããçµæãã·ã¿ã©ãã³ãã¡
ãœããã»ãŒããã¢ãããããªã³ãããªãŠã ãïŒâ
ã¢ã¶ã°ã¢ãã³ãïŒâãã«ãªããŠã©ã·ã«ã®ããšã代
è¬æ®æå€ç³»ã®ç©è³ªãè «çæäœãšçµåãããŠæè «ç
æ§ç©è³ªã補é ããã«é©ããŠããäºãèŠãã ãæ¬çº
æããªãã«è³ã€ãã æ¬çºæã¯ã现èæ¯æ§ã®é«ãäžæ²æè «çæ§ä»£è¬æ®
æç©è³ªã極ããŠç©åãªæ¡ä»¶äžã§è «çæåã«å¯Ÿãã
æäœãšã¢ããçµåïŒâNHCOâïŒã«ãã€ãŠçµåã
ããæ°èŠãªç©è³ªãæå¹æåãšããæè «çå€ã«ä¿ã
ãã®ã§ãã€ãŠãæè «çå¹æã«ããããªãã现èæ¯
æ§ã¯ãåºçºç©è³ªã®ïŒã€ã§ããäžèšæè «çæ§ä»£è¬æ®
æå€ã«ããã¹ãŠæ Œæ®µã«äœãæè «çå€ãæäŸããã
ãšãç®çãšããã ããã«ãæ¬çºæã¯è «çæåã«å¯Ÿããæäœèªèº«ã®
粟補æ段ããã³ããã粟補æäœãã¢ããåºåã¯ã«
ã«ããã·ã«åºãæããäžèšã®æè «çæ§ä»£è¬æ®æç©
質åã³ã¢ããåºåã¯ãã«ã«ããã·ã«åºãå°å ¥ãã
äžèšã®æè «çæ§ä»£è¬æ®æç©è³ªã«ã¢ããçµåïŒâ
NHCOâïŒã«ãã€ãŠçµåãããããšãããªãäžèš
ã®ç©è³ªãæå¹æåãšããæè «çå€ã補é ããæ¹æ³
ãæäŸããããšãç®çãšããã è¿å¹Žãçš®ã ã®æè «çå€ãåºã䜿çšãããŠããã
æãçšåºŠã®å¹æããããŠããããããã®æè «çå€
ãšããŠã·ã¿ã©ãã³ãã¡ãœãã¬ãã»ãŒããã¢ããã
ããªã³ãããªãŠã ãïŒâã¢ã¶ã°ã¢ãã³ãïŒâãã«
ãªããŠã©ã·ã«ã®ããšã代è¬æ®æå€ã䜿çšãããŠã
ããããããã®ç©è³ªã¯ããèªäœäœããé«ã现èæ¯
æ§ãæããŠããŠãæäžããæã«çœè¡çæžå°ãè±
æ¯ãèè žé害çã®å¯äœçšãåããããšãç¥ãããŠ
ããããã®çºã«ãããè¬å€ã®äœ¿çšã«é床ã®ããã®
ãå®æ ã§ããã ãŸããåŸæ¥ããããçš®ã®è «çæåã«å¯Ÿããæäœ
ã補é ãŸãã¯åé¢ããŠããããã®è «çã®æ²»çã«çš
ããè©Šã¿ããªãããŠããããæãŸããæè «çå¹æ
ã¯åŸãããŠããªããããã«ãæè¿è «çæäœã«æè «
çå€ãååŠçã«çµåãããŠåŸãããæ°èŠãªç©è³ªã«
ããæè «çå¹æãæåŸ ããããšãææ¡ãããŠãã
ããäžèšç©è³ªãåŸãããã®ååŠåå¿ã®æ¡ä»¶ãéé ·
ãããããã«ååãªæé·ã¯åŸãããŠããªãããŸ
ãããããã®å®éšã§çšããããæäœã¯å ç«ã°ãã
ãªã³ç»åãŸã§ã®ç²Ÿè£œããè¡ãªãããŠããªãã®ã§ã
äžè¬ã®å ç«ã°ãããªã³ãå«æããŠããŠãçã®æå³
ã§ã¯è «çæäœãšã¯èªãé£ãããããŠæäœãäžè¬ã®
å ç«ã°ãããªã³ã§ããæ ã«äžèšååç©ãæäžãã
ãçäœã«æ£åžžçµç¹ã®é害ãå šèº«çæ£ãŸãã¯ç¡¬çŽãª
ã©ã®ã¢ããã€ã©ãã·ãŒã·ãšãã¯ã®çããäºãå€
ãã æ¬çºæã¯ãåç©ã«æ¥çš®ããè «çã®æåã«å¯Ÿãã
æäœãå«ãæè¡æž ãæ¡åãããã®äžã®å ç«ã°ãã
ãªã³ç»åãååãããããæŽã«ç¹ç°ãªæ段ã§ç²Ÿè£œ
ããããšã«ãã€ãŠãããçŽç²ãªè «çæäœãå ã¥ã
åé¢ãããããæè «çæ§ä»£è¬æ®æå€ãšçµåããã
ããšã«ãããç©åãªåå¿æ¡ä»¶äžã§è£œé ã§ãããã€
äžèšã®ããšãã¢ããã€ã©ãã·ãŒã·ãšãã¯ãçããª
ãæè «çå€ãæäŸãåŸããã®ã§ããã 以äžæ¬çºæã詳ãã説æããã æ¬çºæã®æå¹æåã§ããç©è³ªã¯ãæäœããã³æ
è «çæ§ä»£è¬æ®æå€ã®äž¡è ã«å æ¥ååšããŠããã¢ã
ãåºåã¯ã«ã«ããã·ã«åºãç©åãªåå¿æ¡ä»¶äžã«å
å¿ããããããšã«ããããã®äž¡è ãã¢ããçµåã
ããŠåŸãããããã®éãå¿ èŠã«ããäžèšä»£è¬æ®æ
å€ã«ããã«ã¢ããåºåã¯ã«ã«ããã·ã«åºãå°å ¥ã
ããã®ãæäœãšåå¿ãããããšã«ããäž¡è ãã¢ã
ãçµåãããã æ¬ç©è³ªã®è£œé ã«çšããæè «çæ§ä»£è¬æ®æå€ãšã
ãŠã¯ã·ã¿ã©ãã³ãã¡ãœãã¬ãã»ãŒããã¢ãããã
ãªã³ãããªãŠã ãïŒâã¢ã¶ã°ã¢ãã³ãªãã³ã«ïŒâ
ãã«ãªããŠã©ã·ã«ã奜ãŸããã äžæ²ã®æè «çæ§ä»£è¬æ®æå€ã®å称ã¯äžè¬åã§ã
ã€ãŠãããã®æ§é ã¯æ¬¡ã®ãšããã§ããã ã·ã¿ã©ãã³ïŒ ã¡ãœãã¬ãã»ãŒãïŒ ã¢ãããããªã³ãããªãŠã ïŒ ïŒâã¢ã¶ã°ã¢ãã³ïŒ ïŒâãã«ãªãã©ãŠã·ã«ïŒ ãŸããæ¬ç©è³ªã®åºçºç©è³ªã®ïŒã€ã§ããæäœãšã
ãŠã¯æ²»çãç®çãšããè «çã®æäœãçšããããã è «çãšããŠã¯ã¶ã«ã³ãŒããŒ180ãäœè€èºçãå
ç°èè «ããšãŒãªããçãâ1210çœè¡ç ãâ
388çœè¡ç ãæ¥æ§çœè¡ç ãæªæ§ãªã³ãè «ãçè «ã
èè «ãæªæ§ç¹æ¯äžç®è «ãæ¥æ§éªšé«æ§çœè¡ç ãã¡ã©
ããŒããæ¥æ§ãªã³ãæ§çœè¡ç ã骚é«çããªã©ã®å
çš®ã®è «çããããããã æ¬çºæã§çšããæäœã®è£œé ã¯æ¥æ¬å ç«åŠäŒç·äŒ
èšé²ç¬¬ïŒå·»198é ïŒ1976幎ïŒèšé²ã®æ¹æ³ã«ããå
ã¯DauphinãM.J.ãã®æ¹æ³ãJ.Immunal.113948
ïŒ1974åç §ïŒãã«æºããŠè¡ãªããåè ã¯ããã€ã³ã
ã®ã³ã³ããªãŒãã¢ãžãŠãã³ãïŒComplete
AjuvantïŒãçšããå ãè «ç现èãåç©ã«ç®äžæ³š
å°ããããšã«ãã€ãŠããããå ç«ãæŽã«ç¶ããŠè «
ç现èãéèå 泚å°ããããšã«ããè¿œå å ç«ããŠ
ããããæäœãåŸãæ¹æ³ã§ãããåŸè ã¯è «çæå
ãåç©ã®è ¹è å ã«ïŒãïŒåå埩æäžããŠåç©ãå
ç«ããŠãããããæäœãåŸãæ¹æ³ã§ããã æ¬çºæã§çšããæäœã¯åçš®æäœãç°çš®æäœã®ã
ããã§ãããããåçš®æäœã奜ãŸããã ãã®ããã«ããŠåŸãæäœã¯ãã¢ãã€ããã€ã¯ã
ããã°ã©ãã€ãŒã«ãããã«ã©ã ã®å å¡«å€ã«æ²»çã®
ç®çã®è «çæåãå ã¥ããã ã·ã¢ã³ãçšããŠçµå
ããããŠçœ®ããŠããããã«ã©ã ã«å å¡«ããã«ã©ã
ã«å ç«ã°ãããªã³ç»åãŸã§ç²Ÿè£œããæäœã®æº¶æ¶²ã
æµå ¥ãããããããŠã«ã©ã äžã§æåãšæäœãšãçµ
åãããåŸã«ãç¹æ®ãªæº¶å€ãã«ã©ã ã«æµå ¥ããŠæ
åã»æäœçµåã解ãããŠæäœã®ã¿ã溶åºãããæŽ
ã«ãããéæããŠç²Ÿè£œæäœæ°Žæº¶æ¶²ãåŸããå³ã¡ã
ãããŠåŸãããæäœã¯åŸæ¥ã®å ç«ã°ãããªã³ç»å
ãããæŽã«çŽåºŠã®é«ãæè «çå ç«ã°ãããªã³ã§ã
ãã äžè¬ã«æäœã®ç²Ÿè£œã«ã¯ãç¡«å®å¡©æãšDEAEã»ã«
ããŒã¹ã«ã©ã ã«ããã€ãªã³äº€æã¯ãããã°ã©ãã€
ãŒã䜿ã€ãŠãæè¡æž ããåç»ãåŸãæ¹æ³ããã°ã
ã°çšãããããæ¬çºæã§ã¯ãããã«å ããŠãã¢ã
ã€ããã€ã¯ãããã°ã©ãã€ãŒãçšããŠè «ç现èã«
察ããç¹ç°çæäœã®ã¿ãéžæçã«åŸãçºã®ç¹ç°ç
粟補æäœãè¡ãªã€ãã ã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒã¯ãé µçŽ ãšåº
質ãæäœãšæåãªã©ã®ãããªçäœç©è³ªçžäºéã«å
ãç¹ç°ç芪ååãå©çšããäžæ¹ã®çäœç©è³ªã䜿ã€
ãŠä»æ¹ãéžæçã«åé¢ãããšããåçã«ããšã¥ã
ãã®ã§ããã æ¬çºæã§çšããããã¢ãã€ããã€ã¯ãããã°ã©
ãã€ãŒã«ã¯ã(1)è «ç现èããæœåºããæåãã»ã
ã¢ããŒã¹ïŒSepharoseRïŒã®ããšãæ äœã«ïŒãã
ã ã·ã¢ã³ãçšããŠïŒå ±æçµåããããããã«ã©ã
ã«å å¡«ããæäœæº¶æ¶²ãéããŠãæäœãæåã«çµå
ãããæŽã«ååéã®æº¶åªãæµããŠçµåããªãã€ã
æäœãæŽãå»ã€ãåŸãPHã®ããäœã緩衡溶液ãæµ
ãå ¥ããŠãæäœã»æåã®çµåã解ããŠãåé¢ãã
æäœã溶åºãããæ¹æ³ã(2)ã«ã©ã ãçšãããæå
ãçµåãããæ äœãšæäœæº¶æ¶²ãæ··åããŠæäœãæ
åã«çµåãããæ äœç²åãæŽæµããŠçµåããªãã€
ãæäœãé€ããåŸãæäœã溶é¢ãããæ¹æ³ããã
ã³(3)äžèšã§æåãçµåãããæ äœã®ä»£ãã«ãè «ç
现èèªèº«ãçšããæ¹æ³ãå«ããæ¬çºæã®å®æœäŸã§
ã¯äžèš(1)ãš(3)ã®æ¹æ³ãæ¡çšããã åŸã€ãŠãã¢ãã€ããã€ã¯ãããã°ã©ãã€ã«ãã€
ãŠç²Ÿè£œãããæäœã¯åŸæ¥ã®å ç«ã°ãããªã³ç»åã
ããé¥ã«çŽåºŠã®é«ãæè «çå ç«ã°ãããªã³å³ã¡æ
è «çæäœã§ããã ãã®ããã«ããŠåŸãããæäœãæè «çæ§ä»£è¬æ®
æå€ã«çµåãããã«ã¯ãã¢ããåºåã¯ã«ã«ããã·
ã«åºãæããäžèšä»£è¬æ®æå€ãããã¯ã¢ããåºå
ã¯ã¢ããåºãå°å ¥ããäžèšä»£è¬æ®æå€ãšã氎溶æ§
溶åªäžã«æº¶è§£ããããããã«ã«ã«ããžã€ããã觊
åªãšããŠå ããŠïŒã50âã奜ãŸããã¯10ã40âã§
10åãïŒæéã奜ãŸããã¯30åãïŒæéåå¿ãã
ãŠé ¢é žâé ¢é žãããªãŠã ç·©è¡æ¶²ãªã©ã®æ·»å ã§åå¿
ãåæ¢ãããã 次ã«ãã®åå¿æ¶²äžã®éå°ãªä»£è¬æ®æå€ã觊åªã
ãã³äžèšåå¿åæ¢æ¶²ã®æåãªãã³ã«å¡©é¡ãé€ãçº
ã«éæãã²ã«éãªãã³ã«éå€éã®äœããã®æ
äœãè¡ãªãããåã¯ãããã®æäœãçµã¿åããæ
äœãè¡ãªãã äžèšåå¿ã§è§ŠåªãšããŠçšããã«ã«ããžã€ãã
ã¯ãïŒâãšãã«âïŒâïŒïŒâãžã¡ãã«ã¢ãããã
ãã«ïŒâã«ã«ããžã€ãããïŒâã·ã¯ãããã·ã«â
ïŒâïŒïŒâã¢ã«ããªããšãã«ïŒã«ã«ããžã€ããå
ã¯ãžã·ã¯ãããã·ã«ã«ã«ããžã€ãããå«ãã æè «çæ§ä»£è¬æ®æå€ã«å¿ èŠã«ããã¢ããåºåã¯
ã«ã«ããã·ã«åºãå°å ¥ããã«ã¯ã該æ®æå€ãã®ãŸ
ãŸã«ãåã¯NaããAgã®ããšãå¡©ã®åœ¢äœã«ãã
ãã®ã«ãäžè¬åŒïŒžïŒCH2ïŒoCOOHïŒåŒäžïŒžã¯Clå
ã¯Brã瀺ããïœã¯ïŒä¹è³ïŒã®æŽæ°ã瀺ãïŒã®å
åç©ãæãã¯äžè¬åŒHClã»NH2ïŒCH2ïŒoCOXïŒåŒ
äžïŒžã¯Clåã¯Brã瀺ããïœã¯ïŒä¹è³ïŒã®æŽæ°ã
瀺ãïŒã®ååç©ãæ°Žæ§æº¶åªãäŸãã°ã¡ã¿ããŒã«ã
æ°Žããšã¿ããŒã«ããžã¡ãã«ã¹ã«ããªãã·ãããžãª
ããµã³äžã§ïŒã50âã奜ãŸããã¯10ã40âã§10å
ä¹è³72æéåå¿ããããšãããåå¿çæç©ã¯æ°Žã
ã¢ã«ã³ãŒã«ãã¯ãããã«ã ããžãªããµã³ã®ããšã
溶åªãçšããŠåçµæ¶ããããšã«ããæè «çæ§ä»£è¬
æ®æå€èªå°äœãåŸãããã æ¬ç©è³ªã¯æäœïŒååã«å¯ŸããŠæè «çæ§ä»£è¬æ®æ
å€ã®ïŒã10ååãçµåãããã®ã§ããã æ¬ç©è³ªã®åºä¹³åç©ã«å¯Ÿããæ¥æ§æ¯æ§ã¯ããŠã¹ã
çšã300mgïŒKgã®éèæäžã§èª¿ã¹ããïŒé±éã®èŠ³
å¯ã§æ»äº¡ãèªããããªãã€ãã ãããã€ãŠãæ¬ç©è³ªã¯æ¯æ§ãäœãåçš®ã®äººçã«
察ããŠæå¹ã§ãããäŸãã°ãæ¥æ§çœè¡ç ãæªæ§ãª
ã³ãè «ãçè «ãèè «ãæªæ§ç¹æ¯äžç®è «ãæ¥æ§éªšé«
æ§çœè¡ç ãã¡ã©ããŒããæ¥æ§ãªã³ãæ§çœè¡ç ã骚
é«ççã«æå¹ã§ããã æ¬ç©è³ªãæè «çå€ãšããŠçšããå Žåã®è£œå€åæ¹
æ³ãããã³æäžã®æ¹æ³ãšããŠã¯æè «çå€ã«é¢ãã
å ¬ç¥ã®æ¹æ³ãé©çšãåŸããæäžæ¹æ³ãšããŠã¯çµ
å£ã泚å°ãŸãã¯çŽè žæäžãããããæäžåœ¢æ ãšã
ãŠã¯ç²æ«ãé¡ç²ãé å€ãŸãã¯æ³šå°å€ã座è¬çã®ã
ããã§ãã€ãŠãããã ç¹ã«æ³šå°ã«ããæäžã奜ãŸããã 泚å°è¬ã®è£œå€ã«ã¯çççé£å¡©æ°Žãæ» èæ°Žããªã³
ã²ã«æ¶²çã®æ°Žæº¶æ§æº¶å€ãé氎溶æ§æº¶å€ãç匵å
å€ãç¡çåå€ãå®å®å€ãé²è å€ãæžæ¿åå€ãç·©è¡¡
å€ãä¹³åå€çãä»»æã«çšãåŸãã ãã®äžäŸã瀺ããšãæ¬ç©è³ª10mgãšãã³ãããŒã«
50mgãèžæºæ°Žã«æº¶è§£ããŠ10mlãšããŠåžžæ³ã§é€èã
ãåŸïŒmlå®ã泚å°çšå°ç¶ã«å泚ããåã¯ãã®ãŸãŸ
åçµä¹Ÿç¥ããŠæ³šå°å€ãšããããããŠæ¬å€ã¯äœ¿çšã«
éããçççé£å¡©æ°Žã§çšéããŠæ³šå°æ¶²ãšããã æ¬ç©è³ªã¯è£œå€åäžäžè¬ã«0.01ã90ïŒ å¥œãŸããã¯
0.1ã60ïŒ å«æããããšãã§ããã æ¬ç©è³ªã®æäžéã¯äž»ãšããŠçç¶ã«å·Šå³ãããã
æ人ïŒäººïŒæ¥åœã0.1ãïŒïœã奜ãŸããã¯ïŒãïŒ
ïœã§ããã ãªããæ¬çºæã«ãããšãæäœã®åè «çæ§ãšæè «
çå€ãªãã³ã«æè «çå€ã®æè «çæ§ã¯å€±ãããããš
ãªãäžèšååç©ã«ãã®ãŸãŸä¿ãããŠããã®ã§ãæ¬
ç©è³ªã¯æäžããããšå¹çããç®çãšããè «çéšäœ
ã«å°éããæè «çå¹æãçºæ®ãããåŸã€ãŠæ¬ç©è³ª
ããã®æåãšããŠå«æããæè «çå€ã®ééãåºæº
ãšããŠèãããªãã°ãåäžã®æè «çå€ãã®ãã®ã®
æäžéã®1/10ã1/20ã«çžåœããééã®æè «çå€ã
æåãšããŠå«æããæ¬ç©è³ªã®æäžã«ãã€ãŠåçšåºŠ
ã®è «çå¢æ®æå¶ãåŸããããã€æåãšããŠå«æã
ãæè «çå€ã«ããå¯äœçšã¯ãåäžã®æè «çå€ãã®
ãã®ã®æäžã®å Žåã®1/10ã2/10ã«ãããªããšæåŸ
ããããããã¯æ¬ç©è³ªã®æåããããã®å¥œãŸãã
æ§è³ªã®è€åå¹æãšäºãããšãã§ããã 以äžæ¬çºæãå®æœäŸã«ãã€ãŠæŽã«è©³çŽ°ã«èª¬æã
ãã å®æœäŸ ïŒ ïŒâïŒïŒã¢ãã€ããã€ã¯ãããã°ã©ãã€(3)ãå©çš
ããæäœã®èª¿è£œãšç²Ÿè£œ DBAïŒïŒããŠã¹ãçšããŠç¶ä»£å¹é€ããè ¹
æ°Žåâ388è «ç现èãçççé£å¡©æ°Žã«æžæ¿
ããããã€ããã€ã·ã³ïŒ£ïŒ50ÎŒïœïŒmlïŒãå
ããŠ30åé37âã§åŠçããã®ã¡é å¿åé¢ã«ã
ã€ãŠäžæž ãé€ãã现èã0.85ïŒ ã®ççé£å¡©æ°Ž
ã§ïŒåæŽæ»ãããããããŠå¢æ®èœã倱ã€ã
â388è «ç现èã«ããã€ã³ãïŒFreundïŒã®ã³
ã³ããªãŒãã¢ãžãŠãã³ãïŒComplete
AjuvantïŒïŒFCAãšç¥ãïŒãæ··åããäœé2.9
Kgã®ãŠãµã®è¶³è¹ ã®ç®äžã«ïŒå¹åœã现è108å
ã®å²åã§æ³šå°ããïŒé±éåŸã«åæ§ãªæ¹æ³ã§æ³š
å°ããŠãŠãµã®ãå ç«ããæŽã«ïŒé±éåŸåäžã®
现è108åããã®ãŠãµã®ã®éèã«æ³šå°ããã
ïŒé±éçµéåŸã«é åèã«ã«ããŠãŒã¬ãæ¿å ¥ã
ãŠå šè¡ãæ¡åããããããæè¡æž ãåé¢ããŠ
äžèšã®ç²Ÿè£œãè¡ã€ããå³ã¡ã100mlã®æè¡æž
ã«ç¡«é žã¢ã³ã¢ããŠã ã飜åéã®20ã30ïŒ æ·»å
ããŠçããå¡©æç»åãæ¡åããŠãããã20ml
ã®æ°Žã«å溶解ãããã®æº¶æ¶²ãPH7.0ã®10ïœïŒ
çé žå¡©ç·©è¡é£å¡©æ°Žæº¶æ¶²ïŒPBSãšç¥ãïŒã«å¯Ÿã
ãŠïŒâã72æééæããŠè±å¡©ããïŒãã®éã
24æéæ¯ã«éæå€æ¶²ã®äº€æãè¡ã€ãïŒããã
ã«ãé£å¡©æ°Žã§ïŒåæŽæµããæ£åžžãªDBAïŒïŒ
ããŠã¹ã®è¡çãçéæ··åããïŒâã§30åæŸçœ®
ããŠåžåãè¡ããããåŸãé å¿åé¢ããŠäžæž
ãåŸãããã®åžåæäœãæŽã«ïŒåè¡ã€ãŠãå š
éšã§ïŒåã®åžåæäœãå ããããããŠåŸãã
ãæäœã粟補åæäœãšäºãïŒé垞ïœïŒ§ïŒã
次ãã§äžèšã®æ¹æ³ã§æŽã«ç²Ÿè£œãè¡ã€ããå³ã¡
äžèšåžåã®æžãã äžæž ã«ãâ388现èãç
éæ··åããïŒâã«30åæŸçœ®ããŠïŒ°â388现è
ã«å¯Ÿããæäœãâ388è «ç现èã«çµåãã
ãåŸãé å¿åé¢ããŠäžæž ãé€ããæ²æŸ±ã«PH
3.0ã®ã°ãªã·ã³âå¡©é žç·©è¡æ¶²ãå ããŠãæäœ
ãéé¢ãããããã®æ··åç©ãé å¿åé¢ããŠã
æäœãå«ãäžæž ãæ¡åãããäžæž ã®PHã
0.1Mèæ§ãœãŒã氎溶液ã§äžæ§è¿ãã«èª¿æŽã
ãåŸãPBSã«å¯ŸããŠïŒâã§24æééæãè¡ã€
ãïŒïŒæéæ¯ã«éæå€æ¶²ã亀æããïŒããã
ãŠè¡ããããã®ããŠãµã®ã®æâ388çœè¡ç
è «çå ç«æäœã®æ°Žæº¶æ¶²ã§ããã ïŒâïŒïŒè «ç现èããã³æ£åžžçŽ°èã«å¯Ÿããé害æ§
è©ŠéšïŒãã®ïŒïŒ äžèšã§åŸããŠãµã®ã®æâ388è «çå ç«æ
äœã«ã€ããŠè£äœïŒã¢ã«ã¢ããã®è¡æž ïŒååšäž
ã®çŽ°èé害詊éšãè¡ã€ããå³ã¡ãäžèšæäœæ°Ž
溶液åã¯ããã10ã100ããã³1000åã«çšé
ãããã®ãšãâ388è «ç现èæµ®é液æãã¯
æ£åžžDBAïŒïŒããŠã¹èŸçŽ°èæµ®é液ïŒäœãã
ã€ãŒã°ã«ïŒEagleïŒã®Minimum Essential
MediumïŒä»¥äžMEMãšç¥ãïŒã溶åªãšããŠ
çšãã现èæ¿åºŠã¯ïŒÃ106åïŒãšããïŒãšã
100ÎŒå®æ··åãã宀枩ã§15åæŸçœ®ããŠæäœ
ãããããã®çŽ°èã«çµåããããããã®åŸã¢
ã«ã¢ããã®è¡æž ãã€ãŒã°ã«ã®MEMã§ïŒåã«
çšéãïŒãããè£äœãšäºãïŒãã®100ÎŒ
ããäžèšã®æ··å液ã«å ãã37âã§30åå¹é€
ããé å¿åé¢ããŠãæ²æŸ±ãã€ãŒã°ã«ã®MEM
ã§ïŒåæŽæ»ããããã«ããªãã³ãã«ãŒæ¶²ãå
ããŠãé¡åŸ®é¡äžã§äžèšããããã®çŽ°èã®æ»æ»
ã®çšåºŠã芳å¯ããã ãã®çµæã¯äžèšã®ç¬¬ïŒè¡šã«ç€ºãããšãã现
èã®æ»æ» çšåºŠïŒçŽ°èé害掻æ§ïŒãïŒãïŒïŒãïŒ
ïŒïŒã®ïŒæ®µéã«åããŠè¡šç€ºããïŒæ»æ» ãªãã¯
âã§ç€ºãïŒãšæè¡æž ãäžèšã®æ¹æ³ã§ç²Ÿè£œãã
åŸã«æäœãåãåºãããã®ã¯ãæè¡æž ã粟補
ããããšãªãæäœãåãåºãããã®ã«ããã¹
ãŠïŒ°â388çœè¡ç 现èã«å¯Ÿããæ¯æ§ã¯ããŸã
倧ããç°ããªãããæ£åžžã®DBAïŒïŒããŠã¹
èŸçŽ°èã«å¯Ÿããæ¯æ§ã¯æ¥µããŠäœããããã殺
ãããšã¯ãªãã€ããå³ã¡æè¡æž ã®ç²Ÿè£œã¯ç®ç
ã«ããªã€ãŠããããšããã瀺ãããã
ãè¡šã
ãªããããã§æ£åžžçŽ°èã®ä»£è¡šãšããŠçšãã
DBAïŒïŒããŠã¹ã®èŸçŽ°èã¯ãèŸãæåºãã
åŸããã³ã»ãããçšããŠã€ãŒã°ã«ã®MEMäž
ã§ããã现ãç ãã200meshã®ã¹ãã³ã¬ã¹ç¶²
ãééãããïŒåMEMã§æŽæ»ããããªã¹â
ïŒããããã·ã«ã¡ãã«ïŒã¢ããã¡ã¿ã³ç·©è¡
0.75ïŒ å¡©åã¢ã³ã¢ããŠã 氎溶液ïŒPH7.4ïŒïŒ
mlãå ããŠè©Šæäžã®èµ€è¡çãç Žå£ãæŽã«ã€ãŒ
ã°ã«ã®MEMã§ïŒåæŽã€ãŠåŸããã®ã§ããã ïŒâïŒïŒã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã
æäœã®ç²Ÿè£œ è «çæåãçµåãããã«ã©ã ã䜿çšããã¢
ãã€ããã€ã¯ãããã°ã©ãã€ãŒãçšããŠäžèš
ã«ç€ºãããšã粟補ãè¡ã€ããå ãæåèªèº«ã
äžèšã«ãã粟補ãããå³ã¡DBAïŒïŒããŠã¹
ãçšããŠç¶ä»£å¹é€ããâ388è «ç现èãå
çµä¹Ÿç¥ãããã®ãã®30ïœã«ïŒïœïŒçé žã«ãªãŠ
ã ç·©è¡æ¶²ã§ç·©è¡ãã3MKCl溶液ïŒPH7.4ïŒã
å ããŠãæåã®æœåºã20æéè¡ããæœåºæ¶²ã
65000Gã§10åéé å¿åé¢ããŠäžæž ãæ¡å
ããæŽã«ãã®äžæž ã180000Gã§30åéé å¿å
é¢ããŠäžæž ãæ¡åããèžæºæ°Žã«å¯ŸããŠïŒâã§
70æééæããïŒãã®é24æéæ¯ã«å€æ¶²ã亀
æïŒãéæåŸæŽã«65000Gã§éæ液ãé å¿åé¢
ããŠæ²æŸ±ãé€ããäžæž ã«ç¡«é žã¢ã³ã¢ããŠã ã
å ããŠ2Mã®æ¿åºŠã«ããŠããã65000Gã§10å
éé å¿åé¢ããŠæ²æŸ±ãæ¡åããããã®æ²æŸ±ã
èžæºæ°Žã«æº¶è§£ãããã®æº¶æ¶²ãèžæºæ°Žã«å¯ŸããŠ
72æééæããïŒãã®éã24æéæ¯ã«å€æ¶²ã
亀æããïŒã ããããŠåŸãâ388è «çæåãçšããŠã
ã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒçšã«ã©ã ã
äžèšã®ããšãäœè£œããã å ããã¢ã¬ããŒã¹ã²ã«ïŒSepharose 4BïŒ
Pharmacia Japan Co Ltdã®è£œåïŒãæ°Žã§èš
最ãããŠ20mlãšããããã«ãïŒïœïŒmlã®ãã
ã ã·ã¢ã³æ°Žæº¶æ¶²20mlãå ããŠãPHã11.0ã«ä¿
ã¡ã€ã€äž¡è ãïŒåéåå¿ãããŠããåå¿ç©ã
ã¬ã©ã¹æã§éããæ²æŸ±ãæäžã§æ°·å·ã
ãèžçæ°Žã次ã«æ°·å·ãã0.5Mçé žæ°ŽçŽ ãã
ãªãŠã 氎溶液ã§æŽæ»ããåŸãçŽã¡ã«0.1Mç
é žæ°ŽçŽ ãããªãŠã 氎溶液ã«æžæ¿ãããããã«
äžè¿°ãã粟補æå氎溶液ãæ·»å ãã宀枩ã§ïŒ
å€æ¹æããªããåå¿ãããããçæç©ãã¬ã©
ã¹æã§éããå ã¥0.1Mçé žæ°ŽçŽ ãããª
ãŠã 氎溶液ã次ãã§èžæºæ°ŽãæåŸã«çé žå¡©ç·©
è¡é£å¡©æ°Žæº¶æ¶²ïŒ0.85ïŒ ãPH7.0ïŒã§æŽæ»ã
ãããã®æŽæ»ããåå¿çæç©ãå åŸ13mmé«ã
15cmã®ã¬ã©ã¹ç®¡ã«å å¡«ããŠã¢ãã€ããã€ã¯ã
ããã°ã©ãã€ãŒçšã«ã©ã ãšããããã®ã«ã©ã
ã«åèšïŒâïŒã®æäœïŒãã ãâ388è «ç现
èãšã®çµåã«ããæäœã¯çç¥ïŒã§äœæããæ
äœæº¶æ¶²ïŒïŒ©ïœïŒ§ïŒïŒmlãæµå ¥ãã次ãã§ïŒïœ
ïŒçé žå¡©ç·©è¡é£å¡©æ°Žæº¶æ¶²ïŒ0.85ïŒ ãHz7.0ïŒ
ããã«ã©ã ã®æº¶åºæ¶²ã«èçœãæ€åºãããªããª
ããŸã§æµãã次ãã§ã50ïœïŒã°ãªã·ã³äžå¡©é ž
ç·©è¡æ°Žæº¶æ¶²ïŒPH4.0ïŒãæ·»å ãã0.5Mé£å¡©æ°Ž
溶液ãã«ã©ã ã«æµããŠã溶åºããç»åãæ¡å
ãããããçŽã¡ã«çé žæ°ŽçŽ ãããªãŠã ã§äžæ§
ã«ããçé žå¡©ç·©è¡é£å¡©æ°Žæº¶æ¶²ïŒ0.85ïŒ ãPH
7.0ïŒã«å¯ŸããŠ72æééæããïŒ24æéæ¯ã«
éæå€æ¶²ã亀æããïŒãããããŠã«ã©ã ã«ã
ãã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒãçšããŠ
粟補ããâ388è «çã«å¯Ÿãã粟補æäœæ°Žæº¶
液ãåŸãã ïŒâïŒïŒè «ç现èããã³æ£åžžçŽ°èã«å¯Ÿããé害æ§
è©ŠéšïŒãã®ïŒïŒ äžèšïŒâïŒã§åŸããŠãµã®ã®æâ388è «ç
å ç«æäœã«ã€ããŠïŒâïŒãšåæ§ãªæ¹æ³ã§çŽ°è
é害æ§è©Šéšãè¡ã€ããçµæã¯äžèšç¬¬ïŒè¡šã«ç€º
ãã
DBAïŒïŒããŠã¹ã®èŸçŽ°èã¯ãèŸãæåºãã
åŸããã³ã»ãããçšããŠã€ãŒã°ã«ã®MEMäž
ã§ããã现ãç ãã200meshã®ã¹ãã³ã¬ã¹ç¶²
ãééãããïŒåMEMã§æŽæ»ããããªã¹â
ïŒããããã·ã«ã¡ãã«ïŒã¢ããã¡ã¿ã³ç·©è¡
0.75ïŒ å¡©åã¢ã³ã¢ããŠã 氎溶液ïŒPH7.4ïŒïŒ
mlãå ããŠè©Šæäžã®èµ€è¡çãç Žå£ãæŽã«ã€ãŒ
ã°ã«ã®MEMã§ïŒåæŽã€ãŠåŸããã®ã§ããã ïŒâïŒïŒã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã
æäœã®ç²Ÿè£œ è «çæåãçµåãããã«ã©ã ã䜿çšããã¢
ãã€ããã€ã¯ãããã°ã©ãã€ãŒãçšããŠäžèš
ã«ç€ºãããšã粟補ãè¡ã€ããå ãæåèªèº«ã
äžèšã«ãã粟補ãããå³ã¡DBAïŒïŒããŠã¹
ãçšããŠç¶ä»£å¹é€ããâ388è «ç现èãå
çµä¹Ÿç¥ãããã®ãã®30ïœã«ïŒïœïŒçé žã«ãªãŠ
ã ç·©è¡æ¶²ã§ç·©è¡ãã3MKCl溶液ïŒPH7.4ïŒã
å ããŠãæåã®æœåºã20æéè¡ããæœåºæ¶²ã
65000Gã§10åéé å¿åé¢ããŠäžæž ãæ¡å
ããæŽã«ãã®äžæž ã180000Gã§30åéé å¿å
é¢ããŠäžæž ãæ¡åããèžæºæ°Žã«å¯ŸããŠïŒâã§
70æééæããïŒãã®é24æéæ¯ã«å€æ¶²ã亀
æïŒãéæåŸæŽã«65000Gã§éæ液ãé å¿åé¢
ããŠæ²æŸ±ãé€ããäžæž ã«ç¡«é žã¢ã³ã¢ããŠã ã
å ããŠ2Mã®æ¿åºŠã«ããŠããã65000Gã§10å
éé å¿åé¢ããŠæ²æŸ±ãæ¡åããããã®æ²æŸ±ã
èžæºæ°Žã«æº¶è§£ãããã®æº¶æ¶²ãèžæºæ°Žã«å¯ŸããŠ
72æééæããïŒãã®éã24æéæ¯ã«å€æ¶²ã
亀æããïŒã ããããŠåŸãâ388è «çæåãçšããŠã
ã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒçšã«ã©ã ã
äžèšã®ããšãäœè£œããã å ããã¢ã¬ããŒã¹ã²ã«ïŒSepharose 4BïŒ
Pharmacia Japan Co Ltdã®è£œåïŒãæ°Žã§èš
最ãããŠ20mlãšããããã«ãïŒïœïŒmlã®ãã
ã ã·ã¢ã³æ°Žæº¶æ¶²20mlãå ããŠãPHã11.0ã«ä¿
ã¡ã€ã€äž¡è ãïŒåéåå¿ãããŠããåå¿ç©ã
ã¬ã©ã¹æã§éããæ²æŸ±ãæäžã§æ°·å·ã
ãèžçæ°Žã次ã«æ°·å·ãã0.5Mçé žæ°ŽçŽ ãã
ãªãŠã 氎溶液ã§æŽæ»ããåŸãçŽã¡ã«0.1Mç
é žæ°ŽçŽ ãããªãŠã 氎溶液ã«æžæ¿ãããããã«
äžè¿°ãã粟補æå氎溶液ãæ·»å ãã宀枩ã§ïŒ
å€æ¹æããªããåå¿ãããããçæç©ãã¬ã©
ã¹æã§éããå ã¥0.1Mçé žæ°ŽçŽ ãããª
ãŠã 氎溶液ã次ãã§èžæºæ°ŽãæåŸã«çé žå¡©ç·©
è¡é£å¡©æ°Žæº¶æ¶²ïŒ0.85ïŒ ãPH7.0ïŒã§æŽæ»ã
ãããã®æŽæ»ããåå¿çæç©ãå åŸ13mmé«ã
15cmã®ã¬ã©ã¹ç®¡ã«å å¡«ããŠã¢ãã€ããã€ã¯ã
ããã°ã©ãã€ãŒçšã«ã©ã ãšããããã®ã«ã©ã
ã«åèšïŒâïŒã®æäœïŒãã ãâ388è «ç现
èãšã®çµåã«ããæäœã¯çç¥ïŒã§äœæããæ
äœæº¶æ¶²ïŒïŒ©ïœïŒ§ïŒïŒmlãæµå ¥ãã次ãã§ïŒïœ
ïŒçé žå¡©ç·©è¡é£å¡©æ°Žæº¶æ¶²ïŒ0.85ïŒ ãHz7.0ïŒ
ããã«ã©ã ã®æº¶åºæ¶²ã«èçœãæ€åºãããªããª
ããŸã§æµãã次ãã§ã50ïœïŒã°ãªã·ã³äžå¡©é ž
ç·©è¡æ°Žæº¶æ¶²ïŒPH4.0ïŒãæ·»å ãã0.5Mé£å¡©æ°Ž
溶液ãã«ã©ã ã«æµããŠã溶åºããç»åãæ¡å
ãããããçŽã¡ã«çé žæ°ŽçŽ ãããªãŠã ã§äžæ§
ã«ããçé žå¡©ç·©è¡é£å¡©æ°Žæº¶æ¶²ïŒ0.85ïŒ ãPH
7.0ïŒã«å¯ŸããŠ72æééæããïŒ24æéæ¯ã«
éæå€æ¶²ã亀æããïŒãããããŠã«ã©ã ã«ã
ãã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒãçšããŠ
粟補ããâ388è «çã«å¯Ÿãã粟補æäœæ°Žæº¶
液ãåŸãã ïŒâïŒïŒè «ç现èããã³æ£åžžçŽ°èã«å¯Ÿããé害æ§
è©ŠéšïŒãã®ïŒïŒ äžèšïŒâïŒã§åŸããŠãµã®ã®æâ388è «ç
å ç«æäœã«ã€ããŠïŒâïŒãšåæ§ãªæ¹æ³ã§çŽ°è
é害æ§è©Šéšãè¡ã€ããçµæã¯äžèšç¬¬ïŒè¡šã«ç€º
ãã
ãè¡šã
ãã®çµæã¯ãã¢ãã€ããã€ã¯ãããã°ã©ã
ã€ãŒã«ããæäœã®ç²Ÿè£œã«ãã€ãŠæäœã®ïŒ°â
388现èãžã®æŽ»æ§ãæ Œæ®µã«é«ãŸããèŸçŽ°èã«
察ããæ¯æ§ãäœäžããããšã瀺ããã®ã§ãã¢
ãã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã粟補ã®
åªç§ãã瀺ããŠããã ïŒâïŒïŒæâ388æäœãšæè «çæ§ä»£è¬æ®æå€ãš
ã®çµå åèšïŒâïŒããã³ïŒâïŒã§èª¿è£œããåã ã®
æ¹æ³ã§ç²Ÿè£œãããŠãµã®æâ388æäœãšã·ã¿
ã©ãã³ãã¡ãœãã¬ãã»ãŒããªãã³ã«ïŒâãã«
ãªããŠã©ã·ã«ã®åã ãšãåå¿ããããŠã¢ãã
çµåã«ããããããã®ååç©ãåæããã以
äžãã®åæäŸãè¿°ã¹ãã åæäŸïŒãã®ïŒïŒ æäœãšã·ã¿ã©ãã³ãšã®çµååå¿ ïŒâïŒã§åŸã粟補ãŠãµã®æâ388æäœãïŒml
ã®æ°Žäžã«10.0mgå«æãã氎溶液ã«20.0mgã®ã·ã¿ã©
ãã³ãå ããã®ã¡ãïŒâãšãã«âïŒâïŒïŒâãžã¡
ãã«ã¢ãããããã«ïŒã«ã«ããžã€ããå¡©é žå¡©ã30
mgå ãäžèšã®æéåå¿ããããã®ã«é ¢é žâé ¢é žã
ããªãŠã ç·©è¡æ¶²ïŒPH4.70ïŒïŒmlã®æ·»å ã§åå¿ãå
æ¢ãããã次ãã§åå¿æ¶²ãïŒâã§72æéãïŒã®
èžæºæ°Žã«å¯ŸããŠéæïŒãã®åãå€æ¶²ãïŒå亀æã
ãïŒãããéæå 液ãæ¿çž®ããåŸã«ãããã¹ããª
ã³èªå°äœïŒSephadex â25ãã¢ã«ãã·ã¢ã»ãžã€
ãã³ç€Ÿè£œïŒãå å¡«ããçŽåŸ1.5cmé«ã55cmã®ã«ã©
ã ãéããŠåå¿æ¶²äžã®äœååéç©è³ªãå®å šã«ã«ã©
ã ã«åžçãå»ã溶åºæ¶²ãâ20âã§åçµä¹Ÿç¥ããŠç®
çç©è³ªãåŸããæäœã«å¯Ÿããååå¿æéã«ããã
ã·ã¿ã©ãã³ã®çµåéãBio Assayããæšå®ããçµ
æã第ïŒè¡šã«ç€ºãã
ã€ãŒã«ããæäœã®ç²Ÿè£œã«ãã€ãŠæäœã®ïŒ°â
388现èãžã®æŽ»æ§ãæ Œæ®µã«é«ãŸããèŸçŽ°èã«
察ããæ¯æ§ãäœäžããããšã瀺ããã®ã§ãã¢
ãã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã粟補ã®
åªç§ãã瀺ããŠããã ïŒâïŒïŒæâ388æäœãšæè «çæ§ä»£è¬æ®æå€ãš
ã®çµå åèšïŒâïŒããã³ïŒâïŒã§èª¿è£œããåã ã®
æ¹æ³ã§ç²Ÿè£œãããŠãµã®æâ388æäœãšã·ã¿
ã©ãã³ãã¡ãœãã¬ãã»ãŒããªãã³ã«ïŒâãã«
ãªããŠã©ã·ã«ã®åã ãšãåå¿ããããŠã¢ãã
çµåã«ããããããã®ååç©ãåæããã以
äžãã®åæäŸãè¿°ã¹ãã åæäŸïŒãã®ïŒïŒ æäœãšã·ã¿ã©ãã³ãšã®çµååå¿ ïŒâïŒã§åŸã粟補ãŠãµã®æâ388æäœãïŒml
ã®æ°Žäžã«10.0mgå«æãã氎溶液ã«20.0mgã®ã·ã¿ã©
ãã³ãå ããã®ã¡ãïŒâãšãã«âïŒâïŒïŒâãžã¡
ãã«ã¢ãããããã«ïŒã«ã«ããžã€ããå¡©é žå¡©ã30
mgå ãäžèšã®æéåå¿ããããã®ã«é ¢é žâé ¢é žã
ããªãŠã ç·©è¡æ¶²ïŒPH4.70ïŒïŒmlã®æ·»å ã§åå¿ãå
æ¢ãããã次ãã§åå¿æ¶²ãïŒâã§72æéãïŒã®
èžæºæ°Žã«å¯ŸããŠéæïŒãã®åãå€æ¶²ãïŒå亀æã
ãïŒãããéæå 液ãæ¿çž®ããåŸã«ãããã¹ããª
ã³èªå°äœïŒSephadex â25ãã¢ã«ãã·ã¢ã»ãžã€
ãã³ç€Ÿè£œïŒãå å¡«ããçŽåŸ1.5cmé«ã55cmã®ã«ã©
ã ãéããŠåå¿æ¶²äžã®äœååéç©è³ªãå®å šã«ã«ã©
ã ã«åžçãå»ã溶åºæ¶²ãâ20âã§åçµä¹Ÿç¥ããŠç®
çç©è³ªãåŸããæäœã«å¯Ÿããååå¿æéã«ããã
ã·ã¿ã©ãã³ã®çµåéãBio Assayããæšå®ããçµ
æã第ïŒè¡šã«ç€ºãã
ãè¡šã
åæäŸïŒãã®ïŒïŒ
æäœãšã¡ãœãã¬ãã»ãŒããšã®çµååå¿
ïŒâïŒã§èª¿è£œãåã
ã®æ¹æ³ã§ç²Ÿè£œãããŠãµã®æ
â388æäœãïŒmlã®æ°Žæº¶æ¶²äžã«10mgå«æãã溶
液äžã«6.0mgã®ã¡ãœãã¬ãã»ãŒããå ããŠãæ¹æ
äžã«å¡©é žã§æ¶²ã®PHã4.75ã«èª¿ç¯ãã€ã€2.5mgã®ïŒ
âãšãã«âïŒâïŒïŒâãžã¡ãã«ã¢ãããããã«ïŒâ
ã«ã«ããžã€ããå¡©é žå¡©ãå ããŠäžèšã®æéåå¿ã
ãããã®ã«é ¢é žâé ¢é žãããªãŠã ç·©è¡æ¶²ïŒPH
4.7ïŒïŒmlã®æ·»å ã§åå¿ãåæ¢ããã次ãã§åå¿
液ãïŒâã§72æéïŒã®èžæºæ°Žã«å¯ŸããŠéæïŒã
ã®éãå€æ¶²ãïŒå亀æããïŒãããéæå 液ãæ¿
çž®ããåŸã«ããã¹ããªã³èªå°äœïŒSephadex â
25 ãã¢ã«ãã·ã¢ã»ãžã€ãã³ç€Ÿè£œïŒãå å¡«ããçŽ
åŸ1.5cmé«ã55cmã®ã«ã©ã ãéããŠåå¿æ¶²äžã®äœ
ååéç©è³ªãå®å šã«ã«ã©ã ã«åžçãå»ã溶åºæ¶²ã
â20âã§åçµä¹Ÿç¥ããŠç®çç©è³ªãåŸããæäœã«å¯Ÿ
ããååå¿æéã«ãããã¡ãœãã¬ãã»ãŒãã®çµå
éãäžèšç¬¬ïŒè¡šã«ç€ºãã
â388æäœãïŒmlã®æ°Žæº¶æ¶²äžã«10mgå«æãã溶
液äžã«6.0mgã®ã¡ãœãã¬ãã»ãŒããå ããŠãæ¹æ
äžã«å¡©é žã§æ¶²ã®PHã4.75ã«èª¿ç¯ãã€ã€2.5mgã®ïŒ
âãšãã«âïŒâïŒïŒâãžã¡ãã«ã¢ãããããã«ïŒâ
ã«ã«ããžã€ããå¡©é žå¡©ãå ããŠäžèšã®æéåå¿ã
ãããã®ã«é ¢é žâé ¢é žãããªãŠã ç·©è¡æ¶²ïŒPH
4.7ïŒïŒmlã®æ·»å ã§åå¿ãåæ¢ããã次ãã§åå¿
液ãïŒâã§72æéïŒã®èžæºæ°Žã«å¯ŸããŠéæïŒã
ã®éãå€æ¶²ãïŒå亀æããïŒãããéæå 液ãæ¿
çž®ããåŸã«ããã¹ããªã³èªå°äœïŒSephadex â
25 ãã¢ã«ãã·ã¢ã»ãžã€ãã³ç€Ÿè£œïŒãå å¡«ããçŽ
åŸ1.5cmé«ã55cmã®ã«ã©ã ãéããŠåå¿æ¶²äžã®äœ
ååéç©è³ªãå®å šã«ã«ã©ã ã«åžçãå»ã溶åºæ¶²ã
â20âã§åçµä¹Ÿç¥ããŠç®çç©è³ªãåŸããæäœã«å¯Ÿ
ããååå¿æéã«ãããã¡ãœãã¬ãã»ãŒãã®çµå
éãäžèšç¬¬ïŒè¡šã«ç€ºãã
ãè¡šã
åæäŸïŒãã®ïŒïŒ
ïŒâãã«ãªããŠã©ã·ã«ãšæäœãšã®çµååå¿
ïŒâïŒã«ãã調補ããåã
ã®æ¹æ³ã§ç²Ÿè£œãããŠ
ãµã®æâ388æäœãïŒmlã®æ°Žæº¶æ¶²äžã«10mgãå«
æãã溶液10mläžã«34.7mgã®ïŒâãã«ãªããŠã©ã·
ã«èªå°äœãå ããŠæ¹æäžã«å¡©é žã§æ¶²ã®PHã4.75ã«
調ç¯ãã€ã€ïŒâãšãã«âïŒâïŒâãžã¡ãã«ã¢ãã
ãããã«âã«ã«ããžã€ããå¡©é žå¡©25.2mgãå ããŠ
40ååå¿ãè¡ãªã€ãã次ã«é ¢é žâé ¢é žãããªãŠã
ç·©è¡æ¶²ïŒPH4.75ïŒ20mlã®æ·»å ã§åå¿ãåæ¢ãã
ãã次ãã§ãåå¿æ¶²ãïŒâã§72æéïŒã®PBSã«
察ããŠéæïŒãã®éãå€æ¶²ãïŒå亀æããïŒã
ããéæå 液ãæ¿çž®ããåŸã«ãããã¹ããªã³èªå°
äœïŒSephadex â200ãã¢ã«ãã·ã¢ã»ãžã€ãã³
瀟補ïŒãå å¡«ããçŽåŸïŒcmé«ã65cmã®ã«ã©ã ãé
ããŠåå¿æ¶²äžã®é«ååéç©è³ªåã³äœååéç©è³ªã
å®å šã«åé¢ããã溶åºæ¶²ãè¶ é å¿åé¢æ©ã«ãŠ
40000ïœÃ60åé å¿åé¢ããäžæž ãâ20âã§åçµ
也ç¥ããã ãã®ããã«ããŠåŸãããç©è³ªã§ã¯æäœïŒmgã«å¯Ÿ
ããŠïŒâãã«ãªããŠã©ã·ã«ã¯ã»ãšãã©çµåããŠã
ãªãã€ãã ããã§ïŒâãã«ãªããŠã©ã·ã«ã¯ã¢ãã¯ãã«é ¢é ž
ãåå¿ãããŠã«ã«ããã·ã«åºãå°å ¥ããŠãã®çµå
æ§ãé«ããã以äžãã®äŸãã®ã¹ãã 10mlã®ã¡ã¿ããŒã«ã«500mgã®ïŒâãã«ãªããŠã©
ãžã«ãš86mgã®æ°Žé žåã«ãªãŠã ãå ããããã«ãã
ã«ïŒmlã®èžæºæ°Žãå ãããåŸãããéæ液ã«ãã
ã¡ã«ã145mgã®ã¢ãã¯ãã«é ¢é žãå ããŠ60å宀枩
ã§æ¹æãããåå¿çµäºåŸãæžå§æ¿çž®ãè¡ãªãæ®æž£
ããšãã«ã¢ã³ãŒã«ãšã¯ãããã«ã«ã ããåçµæ¶ã
ããš495mgïŒåç49ïŒ ïŒã®çœè²çµæ¶ãåŸãããã®
ãã®ã¯ãèµ€å€åžåã¹ãã¯ãã«åã³å çŽ åæããäž
èšæ§é ãæããçµåäœã§ããããšã確èªããã
ãµã®æâ388æäœãïŒmlã®æ°Žæº¶æ¶²äžã«10mgãå«
æãã溶液10mläžã«34.7mgã®ïŒâãã«ãªããŠã©ã·
ã«èªå°äœãå ããŠæ¹æäžã«å¡©é žã§æ¶²ã®PHã4.75ã«
調ç¯ãã€ã€ïŒâãšãã«âïŒâïŒâãžã¡ãã«ã¢ãã
ãããã«âã«ã«ããžã€ããå¡©é žå¡©25.2mgãå ããŠ
40ååå¿ãè¡ãªã€ãã次ã«é ¢é žâé ¢é žãããªãŠã
ç·©è¡æ¶²ïŒPH4.75ïŒ20mlã®æ·»å ã§åå¿ãåæ¢ãã
ãã次ãã§ãåå¿æ¶²ãïŒâã§72æéïŒã®PBSã«
察ããŠéæïŒãã®éãå€æ¶²ãïŒå亀æããïŒã
ããéæå 液ãæ¿çž®ããåŸã«ãããã¹ããªã³èªå°
äœïŒSephadex â200ãã¢ã«ãã·ã¢ã»ãžã€ãã³
瀟補ïŒãå å¡«ããçŽåŸïŒcmé«ã65cmã®ã«ã©ã ãé
ããŠåå¿æ¶²äžã®é«ååéç©è³ªåã³äœååéç©è³ªã
å®å šã«åé¢ããã溶åºæ¶²ãè¶ é å¿åé¢æ©ã«ãŠ
40000ïœÃ60åé å¿åé¢ããäžæž ãâ20âã§åçµ
也ç¥ããã ãã®ããã«ããŠåŸãããç©è³ªã§ã¯æäœïŒmgã«å¯Ÿ
ããŠïŒâãã«ãªããŠã©ã·ã«ã¯ã»ãšãã©çµåããŠã
ãªãã€ãã ããã§ïŒâãã«ãªããŠã©ã·ã«ã¯ã¢ãã¯ãã«é ¢é ž
ãåå¿ãããŠã«ã«ããã·ã«åºãå°å ¥ããŠãã®çµå
æ§ãé«ããã以äžãã®äŸãã®ã¹ãã 10mlã®ã¡ã¿ããŒã«ã«500mgã®ïŒâãã«ãªããŠã©
ãžã«ãš86mgã®æ°Žé žåã«ãªãŠã ãå ããããã«ãã
ã«ïŒmlã®èžæºæ°Žãå ãããåŸãããéæ液ã«ãã
ã¡ã«ã145mgã®ã¢ãã¯ãã«é ¢é žãå ããŠ60å宀枩
ã§æ¹æãããåå¿çµäºåŸãæžå§æ¿çž®ãè¡ãªãæ®æž£
ããšãã«ã¢ã³ãŒã«ãšã¯ãããã«ã«ã ããåçµæ¶ã
ããš495mgïŒåç49ïŒ ïŒã®çœè²çµæ¶ãåŸãããã®
ãã®ã¯ãèµ€å€åžåã¹ãã¯ãã«åã³å çŽ åæããäž
èšæ§é ãæããçµåäœã§ããããšã確èªããã
ãåŒã
èç¹160ã165âã§å解
å
çŽ åæçµæ ïŒ
ïŒ
ïŒ
èšç®å€ 27.43ïŒ
1.91ïŒ
10.66ïŒ
å®éšå€ 27.40ïŒ
2.00ïŒ
10.30ïŒ
次ã«ãäžè¿°ã®ããšãããŠåŸãããïŒâãã«ãªã
ãŠã©ã·ã«èªå°äœãæ¬åæäŸã§ããã«è¿°ã¹ãæ¹æ³ãš
ãŸã€ããåãæé ã§ïŒâïŒã§åŸã粟補ãŠãµã®æ
â388现èè «çæäœãšåå¿ãããŠç®çç©è³ªãåŸ
ãã ãã®ç©è³ªã¯æäœïŒmgã«å¯ŸããŠïŒâãã«ãªããŠã©
ã·ã«èªå°äœã10ÎŒïœå«æããŠããã åæäŸïŒãã®ïŒïŒ ïŒâïŒã§åŸã粟補ãŠãµã®æâ388æäœãçš
ããåè¿°ãããšåæ§ãªæé ã§è©²æäœãã·ã¿ã©ã
ã³ãïŒâã¢ã¶ã°ã¢ãã³ãã¡ãœãã¬ãã»ãŒããã¢ã
ããããªã³ãããªãŠã ãªãã³ã«ïŒâãã«ãªããŠã©
ã·ã«èªå°äœïŒåæäŸãã®ïŒåç §ïŒãšããããåå¿
ããããåŸãããåååç©ã¯äžèšåæäŸïŒãã®
ïŒïŒâïŒãã®ïŒïŒã«ããåŸããããã®ãšæ®ãã©åæ§
ãªçµåäœã§ãã€ãã å®æœäŸ ïŒ ïŒâïŒïŒè «ç现èã«å¯Ÿããæäœã®èª¿è£œãšç²Ÿè£œ DBAïŒïŒããŠã¹ãçšããŠç¶ä»£å¹é€ããã
ã€ããã€ã·ã³ïŒ£ã«ãŠå¢æ®èœããªãããè ¹æ°Žå
â388现è107åãïŒé±éæ¯ã«ïŒåäžå¹ã®
DBAïŒïŒè ¹è å ã«æ¥çš®ããïŒåç®ã®æ¥çš®åŸ
ïŒæ¥ç®ã«ãã®ããŠã¹ã麻é ããŠéè ¹ãè ¹éšå€§
éèããæ¡è¡ãããã®è¡æ¶²ããæäœãå«ãæ
è¡æž ã調補ããããããŠ100å¹ã®ããŠã¹ãã
53mlã®æè¡æž ãåŸããæè¡æž ããæäœã®æ¡å
ããã³æäœã®ç²Ÿè£œã¯ïŒâïŒã«æºããŠè¡ãªã€
ãã ïŒâïŒïŒã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã
æäœã®ç²Ÿè£œ DBAïŒïŒãçšããŠç¶ä»£å¹é€ããè ¹æ°Žå
â388è «ç现èãåçµä¹Ÿç¥ãããã®ãã®30ïœ
ã«ïŒïœïŒçé žã«ãªãŠã ç·©è¡æ¶²ã§ç·©è¡ãã
3MKCl溶液ïŒPH7.4ïŒãå ããŠæåã®æœåºã
20æéè¡ãªã€ãããã®æœåºæ¶²ã65000Gã§10
åéé å¿åé¢ããŠäžæž ãæ¡åããæŽã«ãã®äž
æž ã180000Gã§30åé å¿åé¢ããŠäžæž ãæ¡å
ããŠãããèžæºæ°Žã«å¯ŸãïŒâã§72æééæã
ããïŒãã®éã24æéæ¯ã«éæå€æ¶²ã亀æã
ããïŒããããŠåŸãããâ388è «çæåã«ã
ãã¢ãã€ããã€ãŒã¯ãããã°ã©ãã€ãŒã¯æ¬¡ã®
ããšãè¡ãªã€ãã å ããã¢ã¬ããŒã¹ã²ã«ïŒSepharose 4BïŒ
Pharmacia Japan Co Ltdã®è£œåïŒãæ°Žã§èš
最ãããŠ20mlãšããããã«ïŒïœïŒmlã®ããã
ã·ã¢ã³æ°Žæº¶æ¶²20mlãå ããŠãPHã11.0ã«ä¿ã¡
ã€ã€äž¡è ãïŒåéåå¿ãããŠãããåå¿ç©ã
ã¬ã©ã¹æã§éãæ²æŸ±ãæäžã§æ°·å·ãã
èžæºæ°Žæ¬¡ã«æ°·å·ãã0.5Mçé žæ°ŽçŽ ãããªãŠ
ã 氎溶液ã§æŽæ»ããåŸçŽã¡ã«0.1Mçé žæ°ŽçŽ
ãããªãŠã ã«æžæ¿ãããããã«äžè¿°ãã粟補
æå氎溶液ãæ·»å ãã宀枩ã§ïŒå€æ¹æããªã
ãåå¿ãããããçæç©ãã¬ã©ã¹æã§é
ããå ã0.1Mçé žæ°ŽçŽ ãããªãŠã 氎溶液ã
次ãã§èžæºæ°ŽãæåŸã«çé žå¡©ç·©è¡é£å¡©æ°Ž
ïŒ0.85ïŒ PH7.0ïŒã§æŽæ»ããããã®æŽæ»ããå
å¿çæç©ãå åŸ13mmé«ã15cmã®ã¬ã©ã¹ç®¡ã«å
å¡«ããŠã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒçšã«
ã©ã ãšãããåèšïŒâïŒã®æäœã§äœã€ãæè¡
æž ïŒæäœãå«ãïŒæº¶æ¶²ïŒmlãäžèšã®ã¢ãã€ã
ãã€ã¯ãããã°ã©ãã€ãŒçšã«ã©ã ã«æµå ¥ãã
次ãã§ïŒïœïŒçé žå¡©ç·©è¡é£å¡©æ°Žæº¶æ¶²ïŒ0.85ïŒ
PH7.0ïŒããã«ã©ã ã®æº¶åºæ¶²ã«èçœãæ€åºã
ããªããªããŸã§æµããã次ã«50ïœïŒã°ãªã·ã³
âå¡©é žç·©è¡æ°Žæº¶æ¶²ïŒPH4.0ïŒãæ·»å ãã0.5M
é£å¡©æ°Žæº¶æ¶²ãã«ã©ã ã«æµããŠæº¶åºããç»åã
æ¡åããããããçŽã¡ã«çé žæ°ŽçŽ ãããªãŠã
ã§äžæ§ã«ããŠçé žå¡©ç·©è¡é£å¡©æ°Žæº¶æ¶²ïŒ0.85ïŒ
PH7.0ïŒã«å¯ŸããŠ72æééæãããïŒ24æéæ¯
ã«éæå€æ¶²ã亀æãããïŒããããŠã¢ãã€ã
ãã€ã¯ãããã°ã©ãã€ãŒãçšããŠç²Ÿè£œããæ
â388è «ç现èã«å¯Ÿããæäœæ°Žæº¶æ¶²ãåŸ
ãã ïŒâïŒïŒè «ç现èããã³æ£åžžçŽ°èã«å¯Ÿããé害æ§
è©Šéš äžèšïŒâïŒã§åŸãããŠã¹ã®æâ388è «ç
å ç«æäœã«ã€ããŠå®æœäŸïŒãšåæ§ãªæ¹æ³ã§çŽ°
èé害æ§è©Šéšãè¡ãªã€ããçµæã¯äžèšç¬¬ïŒè¡š
ã«ç€ºãã
ãŠã©ã·ã«èªå°äœãæ¬åæäŸã§ããã«è¿°ã¹ãæ¹æ³ãš
ãŸã€ããåãæé ã§ïŒâïŒã§åŸã粟補ãŠãµã®æ
â388现èè «çæäœãšåå¿ãããŠç®çç©è³ªãåŸ
ãã ãã®ç©è³ªã¯æäœïŒmgã«å¯ŸããŠïŒâãã«ãªããŠã©
ã·ã«èªå°äœã10ÎŒïœå«æããŠããã åæäŸïŒãã®ïŒïŒ ïŒâïŒã§åŸã粟補ãŠãµã®æâ388æäœãçš
ããåè¿°ãããšåæ§ãªæé ã§è©²æäœãã·ã¿ã©ã
ã³ãïŒâã¢ã¶ã°ã¢ãã³ãã¡ãœãã¬ãã»ãŒããã¢ã
ããããªã³ãããªãŠã ãªãã³ã«ïŒâãã«ãªããŠã©
ã·ã«èªå°äœïŒåæäŸãã®ïŒåç §ïŒãšããããåå¿
ããããåŸãããåååç©ã¯äžèšåæäŸïŒãã®
ïŒïŒâïŒãã®ïŒïŒã«ããåŸããããã®ãšæ®ãã©åæ§
ãªçµåäœã§ãã€ãã å®æœäŸ ïŒ ïŒâïŒïŒè «ç现èã«å¯Ÿããæäœã®èª¿è£œãšç²Ÿè£œ DBAïŒïŒããŠã¹ãçšããŠç¶ä»£å¹é€ããã
ã€ããã€ã·ã³ïŒ£ã«ãŠå¢æ®èœããªãããè ¹æ°Žå
â388现è107åãïŒé±éæ¯ã«ïŒåäžå¹ã®
DBAïŒïŒè ¹è å ã«æ¥çš®ããïŒåç®ã®æ¥çš®åŸ
ïŒæ¥ç®ã«ãã®ããŠã¹ã麻é ããŠéè ¹ãè ¹éšå€§
éèããæ¡è¡ãããã®è¡æ¶²ããæäœãå«ãæ
è¡æž ã調補ããããããŠ100å¹ã®ããŠã¹ãã
53mlã®æè¡æž ãåŸããæè¡æž ããæäœã®æ¡å
ããã³æäœã®ç²Ÿè£œã¯ïŒâïŒã«æºããŠè¡ãªã€
ãã ïŒâïŒïŒã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã
æäœã®ç²Ÿè£œ DBAïŒïŒãçšããŠç¶ä»£å¹é€ããè ¹æ°Žå
â388è «ç现èãåçµä¹Ÿç¥ãããã®ãã®30ïœ
ã«ïŒïœïŒçé žã«ãªãŠã ç·©è¡æ¶²ã§ç·©è¡ãã
3MKCl溶液ïŒPH7.4ïŒãå ããŠæåã®æœåºã
20æéè¡ãªã€ãããã®æœåºæ¶²ã65000Gã§10
åéé å¿åé¢ããŠäžæž ãæ¡åããæŽã«ãã®äž
æž ã180000Gã§30åé å¿åé¢ããŠäžæž ãæ¡å
ããŠãããèžæºæ°Žã«å¯ŸãïŒâã§72æééæã
ããïŒãã®éã24æéæ¯ã«éæå€æ¶²ã亀æã
ããïŒããããŠåŸãããâ388è «çæåã«ã
ãã¢ãã€ããã€ãŒã¯ãããã°ã©ãã€ãŒã¯æ¬¡ã®
ããšãè¡ãªã€ãã å ããã¢ã¬ããŒã¹ã²ã«ïŒSepharose 4BïŒ
Pharmacia Japan Co Ltdã®è£œåïŒãæ°Žã§èš
最ãããŠ20mlãšããããã«ïŒïœïŒmlã®ããã
ã·ã¢ã³æ°Žæº¶æ¶²20mlãå ããŠãPHã11.0ã«ä¿ã¡
ã€ã€äž¡è ãïŒåéåå¿ãããŠãããåå¿ç©ã
ã¬ã©ã¹æã§éãæ²æŸ±ãæäžã§æ°·å·ãã
èžæºæ°Žæ¬¡ã«æ°·å·ãã0.5Mçé žæ°ŽçŽ ãããªãŠ
ã 氎溶液ã§æŽæ»ããåŸçŽã¡ã«0.1Mçé žæ°ŽçŽ
ãããªãŠã ã«æžæ¿ãããããã«äžè¿°ãã粟補
æå氎溶液ãæ·»å ãã宀枩ã§ïŒå€æ¹æããªã
ãåå¿ãããããçæç©ãã¬ã©ã¹æã§é
ããå ã0.1Mçé žæ°ŽçŽ ãããªãŠã 氎溶液ã
次ãã§èžæºæ°ŽãæåŸã«çé žå¡©ç·©è¡é£å¡©æ°Ž
ïŒ0.85ïŒ PH7.0ïŒã§æŽæ»ããããã®æŽæ»ããå
å¿çæç©ãå åŸ13mmé«ã15cmã®ã¬ã©ã¹ç®¡ã«å
å¡«ããŠã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒçšã«
ã©ã ãšãããåèšïŒâïŒã®æäœã§äœã€ãæè¡
æž ïŒæäœãå«ãïŒæº¶æ¶²ïŒmlãäžèšã®ã¢ãã€ã
ãã€ã¯ãããã°ã©ãã€ãŒçšã«ã©ã ã«æµå ¥ãã
次ãã§ïŒïœïŒçé žå¡©ç·©è¡é£å¡©æ°Žæº¶æ¶²ïŒ0.85ïŒ
PH7.0ïŒããã«ã©ã ã®æº¶åºæ¶²ã«èçœãæ€åºã
ããªããªããŸã§æµããã次ã«50ïœïŒã°ãªã·ã³
âå¡©é žç·©è¡æ°Žæº¶æ¶²ïŒPH4.0ïŒãæ·»å ãã0.5M
é£å¡©æ°Žæº¶æ¶²ãã«ã©ã ã«æµããŠæº¶åºããç»åã
æ¡åããããããçŽã¡ã«çé žæ°ŽçŽ ãããªãŠã
ã§äžæ§ã«ããŠçé žå¡©ç·©è¡é£å¡©æ°Žæº¶æ¶²ïŒ0.85ïŒ
PH7.0ïŒã«å¯ŸããŠ72æééæãããïŒ24æéæ¯
ã«éæå€æ¶²ã亀æãããïŒããããŠã¢ãã€ã
ãã€ã¯ãããã°ã©ãã€ãŒãçšããŠç²Ÿè£œããæ
â388è «ç现èã«å¯Ÿããæäœæ°Žæº¶æ¶²ãåŸ
ãã ïŒâïŒïŒè «ç现èããã³æ£åžžçŽ°èã«å¯Ÿããé害æ§
è©Šéš äžèšïŒâïŒã§åŸãããŠã¹ã®æâ388è «ç
å ç«æäœã«ã€ããŠå®æœäŸïŒãšåæ§ãªæ¹æ³ã§çŽ°
èé害æ§è©Šéšãè¡ãªã€ããçµæã¯äžèšç¬¬ïŒè¡š
ã«ç€ºãã
ãè¡šã
第ïŒè¡šããåãããšãåè¿°ãšåæ§ã¢ãã€ã
ãã€ã¯ãããã°ã©ãã€ãŒã«ãã€ãŠïŒ°â388现
èã«å¯Ÿãã掻æ§ã¯æ Œæ®µã«å¢å€§ããŠããã ïŒâïŒïŒããŠã¹æâ388æäœãšæè «çæ§ä»£è¬æ®
æå€ãšã®çµå ïŒâïŒããã³ïŒâïŒã§åŸãããŠã¹æâ
388æäœãšã·ã¿ã©ãã³ãã¡ãœãã¬ãã»ãŒãã
ã¢ãããããªã³ãããªãŠã ãïŒâã¢ã¶ã°ã¢ã
ã³ãªãã³ã«ïŒâãã«ãªããŠã©ã·ã«ã®åã ãå
èšïŒâïŒã§è¿°ã¹ãæ¹æ³ãšãŸã€ããåæ§ã®æ¹æ³
ã§çµåããããŠãäž¡è ãã¢ããåºã§çµåãã
ç©è³ªãåŸãã ãããã®ç©è³ªã¯äœããåèšïŒâïŒã§åŸã察
å¿ç©è³ªãšæ®ã©åäžã®ç©ççãååŠçæ§è³ªã瀺
ããã å®æœäŸ ïŒ ããŠã¹çœè¡ç â388ã«å¯Ÿããæè «çå¹æ
DBAïŒïŒããŠã¹ãçšããŠç¶ä»£å¹é€ããè ¹æ°Žå
â388现èã10å¹ãããªã矀ã®åDBAïŒïŒããŠã¹
ã®è ¹è å ã«ïŒÃ106åïŒå¹ç§»æ€ãã移æ€ã®24æé
åŸããåçš®æäœãåçš®åžè²©æè «çå€ããããåç¬
ããã³ãŠãµã®æâ388æäœãšåçš®åžè²©æè «çå€
ãšã®ååç©ããã³ããŠã¹æâ388æäœãšåçš®åž
販æè «çå€ãšã®ååç©ã®ããããã®æ°Žæº¶æ¶²ãïŒæ¥
ïŒåïŒæ¥éé£ç¶ãåèšã§ïŒååããŠã¹ã®è ¹è å ã«
泚å°ããè©ŠææäžçŸ€ã®å¹³åçåæ¥æ°ïŒïŒŽïŒããã³
å¯Ÿç §çŸ€ã®å¹³åçåæ¥æ°(C)ãæ±ãã延åœçïŒïŒŽïŒïŒ£
Ã100ïŒãç®åºããã çµæã第ïŒè¡šä¹è³ç¬¬10è¡šã«ç€ºãã
ãã€ã¯ãããã°ã©ãã€ãŒã«ãã€ãŠïŒ°â388现
èã«å¯Ÿãã掻æ§ã¯æ Œæ®µã«å¢å€§ããŠããã ïŒâïŒïŒããŠã¹æâ388æäœãšæè «çæ§ä»£è¬æ®
æå€ãšã®çµå ïŒâïŒããã³ïŒâïŒã§åŸãããŠã¹æâ
388æäœãšã·ã¿ã©ãã³ãã¡ãœãã¬ãã»ãŒãã
ã¢ãããããªã³ãããªãŠã ãïŒâã¢ã¶ã°ã¢ã
ã³ãªãã³ã«ïŒâãã«ãªããŠã©ã·ã«ã®åã ãå
èšïŒâïŒã§è¿°ã¹ãæ¹æ³ãšãŸã€ããåæ§ã®æ¹æ³
ã§çµåããããŠãäž¡è ãã¢ããåºã§çµåãã
ç©è³ªãåŸãã ãããã®ç©è³ªã¯äœããåèšïŒâïŒã§åŸã察
å¿ç©è³ªãšæ®ã©åäžã®ç©ççãååŠçæ§è³ªã瀺
ããã å®æœäŸ ïŒ ããŠã¹çœè¡ç â388ã«å¯Ÿããæè «çå¹æ
DBAïŒïŒããŠã¹ãçšããŠç¶ä»£å¹é€ããè ¹æ°Žå
â388现èã10å¹ãããªã矀ã®åDBAïŒïŒããŠã¹
ã®è ¹è å ã«ïŒÃ106åïŒå¹ç§»æ€ãã移æ€ã®24æé
åŸããåçš®æäœãåçš®åžè²©æè «çå€ããããåç¬
ããã³ãŠãµã®æâ388æäœãšåçš®åžè²©æè «çå€
ãšã®ååç©ããã³ããŠã¹æâ388æäœãšåçš®åž
販æè «çå€ãšã®ååç©ã®ããããã®æ°Žæº¶æ¶²ãïŒæ¥
ïŒåïŒæ¥éé£ç¶ãåèšã§ïŒååããŠã¹ã®è ¹è å ã«
泚å°ããè©ŠææäžçŸ€ã®å¹³åçåæ¥æ°ïŒïŒŽïŒããã³
å¯Ÿç §çŸ€ã®å¹³åçåæ¥æ°(C)ãæ±ãã延åœçïŒïŒŽïŒïŒ£
Ã100ïŒãç®åºããã çµæã第ïŒè¡šä¹è³ç¬¬10è¡šã«ç€ºãã
ãè¡šã
ãè¡šã
ãè¡šã
ãè¡šã
ãè¡šã
ãè¡šããã
ãè¡šã
ãæäœãè¡šããã
äžèšç¬¬ïŒã10è¡šã§å€ãããšããæ¬ç©è³ªã®ïŒ°â
388现èã«å¯ŸããããŠã¹ã®å»¶åœçã¯ãæ姿ã§ã¯ã
åžè²©ã®æè «çå€ã®ïŒã10åã®æäžéã§ãã»ãŒåžè²©
æè «çå€ã®æå¶çãšçããããã®ããšã¯ç¬¬10è¡šã
ããå€ãããšãããã®çšåºŠã®æäžéã§ã¯æäœèªèº«
ã®è «çæå¶åãã»ãšãã©çŸããªãããšã瀺ããã®
ã§ãã€ãŠãåœç¶ã®ããšã§ãããæ¬ç©è³ªã®ç¹åŸŽã¯ã
æ¬ç©è³ªã®æåã§ããåžè²©æè «çå€ã®æäžéãšãæ¬
ç©è³ªæäžã«ããåžè²©æè «çå€æåã®æäžéãæ¯èŒ
ãããšå€ç¶ãšããã®ã§ãã€ãŠãåŸè ã¯åè ã®1/10
ã1/20ã«éããªãã®ã«ãæ®ãã©åè ãšåçšåºŠã®è «
çå¢æ®æå¶çã瀺ããŠããããã®äºã¯æ¬ç©è³ªã®æ
åã®ïŒã€ã§ããæäœããä»ã®æåã§ããå°éåžè²©
æè «çå€ãåŠäœã«è¯ãè «çéšäœã«å°éããããŠã
ããã瀺ããã®ã«å€ãªãããæ¬çºæã®çæ³ãã¿ã
ãšã«å ·äœåããŠããã æ¬ç©è³ªã¯ãã®äœçšã«ãã€ãŠãå æ¥å¯äœçšã®æ¥µåºŠ
ã«é«ãåžè²©æè «çå€ã®äœ¿çšãåŸæ¥ã®1/10ã1/20ã«
äœäžããªããããã€åŸæ¥ãšåçšåºŠã®è «çå¢æ®æå¶
åãçºæ®ããã®ã§ããã å°ç¹èšãã¹ãã¯ãè «çå¢æ®æå¶çã®ç¹ã§ã¯æäœ
ã®ç±æ¥ã¯é¢ä¿ãªããããŠãµã®ãçšããŠèª¿è£œã粟補
ããæäœãšåžè²©æè «çå€ãšã®çµåããç©è³ªãããŠ
ã¹ã«æäžããå Žåã«ã10å¹ã®ããŠã¹ã®å ïŒå¹çšåºŠ
ã«å šèº«çæ£ããã³ç¡¬çŽãªã©ã®ã¢ããã©ãã·âã·ãš
ãã¯ãåãããããã«å¯ŸãããŠãµã®ãçšããŠèª¿è£œ
ããæ¬çºæã«ããã¢ãã€ããã€ã¯ãããã°ã©ãã€
ãŒã§ç²Ÿè£œããæäœãçšããŠåæããæ¬ç©è³ªãæäž
ããå Žåã«ã¯ããã®ãããªã¢ããã©ãã·âã·ãšã
ã¯ã®çºçã¯å€§ãã«è»œæžãããã ããŠã¹ãçšããŠäœæã»ç²Ÿè£œããæäœãçšããŠå
æããç©è³ªã®å Žåã«ã¯ãã¢ããã©ãã·âã·ãšãã¯
ã®çºçã¯æ¥µããŠçšã§ãã€ãããæŽã«ãã®æäœã®ç²Ÿ
補ãã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã€ãŠè¡
ã€ãå Žåã®æ¬ç©è³ªã§ã¯ã¢ããã©ãã·âã·ãšãã¯ã®
çºçã¯å šãèŠãããªãã€ãã å®æœäŸ ïŒ ïŒâïŒïŒè «ç现èã«å¯Ÿããæäœã®èª¿è£œãšç²Ÿè£œ çŽè žçæ£è ïŒ50æç·æ§ïŒããè¡æ¶²50mlãæ¡
è¡ãããã®è¡æ¶²ããæäœãå«ãæè¡æž 22mlã
åŸãã ïŒâïŒïŒã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã
æäœã®ç²Ÿè£œ æè¡æã«æåºããæ£è è «ç现èãåçµä¹Ÿç¥
ããããã«50mlã®ïŒïœïŒãªã³é žã«ãªãŠã ç·©è¡
液ã§ç·©è¡ãã3MKCl溶液ïŒPH7.4ïŒãå ã
ãŠãæåã®æœåºã20æéè¡ãªã€ãããã®æœåº
液ãã65000Gã§10åéé å¿åé¢ããŠãäžæž
ãæ¡åããŠãããèžçæ°Žã«å¯ŸãïŒâã§72æé
éæãããïŒãã®é24æéæ¯ã«éæå€æ¶²ã亀
æãããïŒããããŠåŸãããæ£è è «çæåã«
ããã¢ãã€ããã€ãŒã¯ãããã°ã©ãã€ãŒã¯æ¬¡
ã®ããšãè¡ãªã€ãã å ãã¢ã¬ããŒã¹ã²ã«ïŒSepharose 4BïŒ
Pharmacia Japan Co Ltdã®è£œåïŒãæ°Žã§èš
最ãããŠ20mlãšããããã«ïŒïœïŒmlã®ããã
ã·ã¢ã³æ°Žæº¶æ¶²20mlãå ããŠPHã11.0ã«ä¿ã¡ã€
ã€äž¡è ãïŒåéåå¿ãããŠãããåå¿çæç©
ãã¬ã©ã¹æã§éããæ²æŸ±ãæäžã§æ°·å·
ããèžçæ°Žã§æŽæ»ãã次ã«æ°Žå·ãã0.5çé ž
æ°ŽçŽ ãããªãŠã 氎溶液ã§æŽæ»ããåŸãçŽã¡ã«
0.1Mçé žæ°ŽçŽ ãããªãŠã ã«æžæ¿ããããã
ã«äžè¿°ãã粟補æå氎溶液ãæ·»å ãã宀枩ã§
ïŒå€æ¹æããªããåå¿ããããã åŸãããåå¿çæç©ãã¬ã©ã¹æã§é
ããããã«å ã0.1Mçé žæ°ŽçŽ ãããªãŠã æ°Ž
溶液ãã次ãã§èžçæ°ŽãæåŸã«çé žå¡©ç·©è¡é£
å¡©æ°ŽïŒ0.85ïŒ PH7.0ïŒã§ããããæŽæ»ããã
ãã®æŽæ»ããåå¿çæç©ãå åŸ13mmé«ã15cm
ã®ã¬ã©ã¹ç®¡ã«å å¡«ããŠã¢ãã€ããã€ã¯ããã
ã°ã©ãã€ãŒçšã«ã©ã ãšãããåèšïŒâïŒã®æ
äœã§èª¿è£œããæè¡æž ïŒæäœãå«ãïŒæº¶æ¶²ïŒml
ãäžèšã®ã¢ãã€ããã€ã¯ãããã°ã©ãã€çšã«
ã©ã ã«æµå ¥ãã次ãã§ïŒïœïŒçé žå¡©ç·©è¡é£å¡©
氎溶液ïŒ0.85ïŒ PH7.0ïŒããã«ã©ã ã®æº¶åºæ¶²
ã«èçœãæ€åºãããªããªããŸã§æµããã次ã«
50ïœïŒã°ãªã·ã³âå¡©é žç·©è¡æ°Žæº¶æ¶²ïŒPH4.0ïŒ
ãæ·»å ãã0.5Mé£å¡©æ°Žæº¶æ¶²ãã«ã©ã ã«æµã
ãŠæº¶åºããç»åãæ¡åããããããçŽã¡ã«ç
é žæ°ŽçŽ ãããªãŠã ã§äžæ§ã«ããŠçé žå¡©ç·©è¡é£
塩氎溶液ïŒ0.85ïŒ PH7.0ïŒå¯ŸããŠ72æééæ
ãããïŒ24æéæ¯ã«éæå€æ¶²ã亀æãããïŒã
ãããŠã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒãçš
ããŠç²Ÿè£œããæ£è è «çã«å¯Ÿããæäœæ°Žæº¶æ¶²ã
åŸãã ãã®ããã«ããŠåŸãæäœã¯ãæ°Žã«å¯æº¶ã§ã
ãããããããã¡ã¿ããŒã«ããšã¿ããŒã«ãã¢
ã»ãã³ããã³ãŒã³çã®ææ©æº¶åªã«ã¯äžæº¶ã§ã
ã€ããããã«ãã®æäœã¯æ·»éå³é¢ã®ç¬¬ïŒå³ã«
瀺ããããªèµ€å€ç·åžåãšç¬¬ïŒå³ã«ç€ºã玫å€ç·
åžåã瀺ããååéã¯ãçŽ15äžã§ãdiscé»æ°
æ³³åã§ã¯Rf0ã0.1ã®éã«ååšããç©è³ªã§ãã€
ãã
äžèšç¬¬ïŒã10è¡šã§å€ãããšããæ¬ç©è³ªã®ïŒ°â
388现èã«å¯ŸããããŠã¹ã®å»¶åœçã¯ãæ姿ã§ã¯ã
åžè²©ã®æè «çå€ã®ïŒã10åã®æäžéã§ãã»ãŒåžè²©
æè «çå€ã®æå¶çãšçããããã®ããšã¯ç¬¬10è¡šã
ããå€ãããšãããã®çšåºŠã®æäžéã§ã¯æäœèªèº«
ã®è «çæå¶åãã»ãšãã©çŸããªãããšã瀺ããã®
ã§ãã€ãŠãåœç¶ã®ããšã§ãããæ¬ç©è³ªã®ç¹åŸŽã¯ã
æ¬ç©è³ªã®æåã§ããåžè²©æè «çå€ã®æäžéãšãæ¬
ç©è³ªæäžã«ããåžè²©æè «çå€æåã®æäžéãæ¯èŒ
ãããšå€ç¶ãšããã®ã§ãã€ãŠãåŸè ã¯åè ã®1/10
ã1/20ã«éããªãã®ã«ãæ®ãã©åè ãšåçšåºŠã®è «
çå¢æ®æå¶çã瀺ããŠããããã®äºã¯æ¬ç©è³ªã®æ
åã®ïŒã€ã§ããæäœããä»ã®æåã§ããå°éåžè²©
æè «çå€ãåŠäœã«è¯ãè «çéšäœã«å°éããããŠã
ããã瀺ããã®ã«å€ãªãããæ¬çºæã®çæ³ãã¿ã
ãšã«å ·äœåããŠããã æ¬ç©è³ªã¯ãã®äœçšã«ãã€ãŠãå æ¥å¯äœçšã®æ¥µåºŠ
ã«é«ãåžè²©æè «çå€ã®äœ¿çšãåŸæ¥ã®1/10ã1/20ã«
äœäžããªããããã€åŸæ¥ãšåçšåºŠã®è «çå¢æ®æå¶
åãçºæ®ããã®ã§ããã å°ç¹èšãã¹ãã¯ãè «çå¢æ®æå¶çã®ç¹ã§ã¯æäœ
ã®ç±æ¥ã¯é¢ä¿ãªããããŠãµã®ãçšããŠèª¿è£œã粟補
ããæäœãšåžè²©æè «çå€ãšã®çµåããç©è³ªãããŠ
ã¹ã«æäžããå Žåã«ã10å¹ã®ããŠã¹ã®å ïŒå¹çšåºŠ
ã«å šèº«çæ£ããã³ç¡¬çŽãªã©ã®ã¢ããã©ãã·âã·ãš
ãã¯ãåãããããã«å¯ŸãããŠãµã®ãçšããŠèª¿è£œ
ããæ¬çºæã«ããã¢ãã€ããã€ã¯ãããã°ã©ãã€
ãŒã§ç²Ÿè£œããæäœãçšããŠåæããæ¬ç©è³ªãæäž
ããå Žåã«ã¯ããã®ãããªã¢ããã©ãã·âã·ãšã
ã¯ã®çºçã¯å€§ãã«è»œæžãããã ããŠã¹ãçšããŠäœæã»ç²Ÿè£œããæäœãçšããŠå
æããç©è³ªã®å Žåã«ã¯ãã¢ããã©ãã·âã·ãšãã¯
ã®çºçã¯æ¥µããŠçšã§ãã€ãããæŽã«ãã®æäœã®ç²Ÿ
補ãã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã€ãŠè¡
ã€ãå Žåã®æ¬ç©è³ªã§ã¯ã¢ããã©ãã·âã·ãšãã¯ã®
çºçã¯å šãèŠãããªãã€ãã å®æœäŸ ïŒ ïŒâïŒïŒè «ç现èã«å¯Ÿããæäœã®èª¿è£œãšç²Ÿè£œ çŽè žçæ£è ïŒ50æç·æ§ïŒããè¡æ¶²50mlãæ¡
è¡ãããã®è¡æ¶²ããæäœãå«ãæè¡æž 22mlã
åŸãã ïŒâïŒïŒã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã
æäœã®ç²Ÿè£œ æè¡æã«æåºããæ£è è «ç现èãåçµä¹Ÿç¥
ããããã«50mlã®ïŒïœïŒãªã³é žã«ãªãŠã ç·©è¡
液ã§ç·©è¡ãã3MKCl溶液ïŒPH7.4ïŒãå ã
ãŠãæåã®æœåºã20æéè¡ãªã€ãããã®æœåº
液ãã65000Gã§10åéé å¿åé¢ããŠãäžæž
ãæ¡åããŠãããèžçæ°Žã«å¯ŸãïŒâã§72æé
éæãããïŒãã®é24æéæ¯ã«éæå€æ¶²ã亀
æãããïŒããããŠåŸãããæ£è è «çæåã«
ããã¢ãã€ããã€ãŒã¯ãããã°ã©ãã€ãŒã¯æ¬¡
ã®ããšãè¡ãªã€ãã å ãã¢ã¬ããŒã¹ã²ã«ïŒSepharose 4BïŒ
Pharmacia Japan Co Ltdã®è£œåïŒãæ°Žã§èš
最ãããŠ20mlãšããããã«ïŒïœïŒmlã®ããã
ã·ã¢ã³æ°Žæº¶æ¶²20mlãå ããŠPHã11.0ã«ä¿ã¡ã€
ã€äž¡è ãïŒåéåå¿ãããŠãããåå¿çæç©
ãã¬ã©ã¹æã§éããæ²æŸ±ãæäžã§æ°·å·
ããèžçæ°Žã§æŽæ»ãã次ã«æ°Žå·ãã0.5çé ž
æ°ŽçŽ ãããªãŠã 氎溶液ã§æŽæ»ããåŸãçŽã¡ã«
0.1Mçé žæ°ŽçŽ ãããªãŠã ã«æžæ¿ããããã
ã«äžè¿°ãã粟補æå氎溶液ãæ·»å ãã宀枩ã§
ïŒå€æ¹æããªããåå¿ããããã åŸãããåå¿çæç©ãã¬ã©ã¹æã§é
ããããã«å ã0.1Mçé žæ°ŽçŽ ãããªãŠã æ°Ž
溶液ãã次ãã§èžçæ°ŽãæåŸã«çé žå¡©ç·©è¡é£
å¡©æ°ŽïŒ0.85ïŒ PH7.0ïŒã§ããããæŽæ»ããã
ãã®æŽæ»ããåå¿çæç©ãå åŸ13mmé«ã15cm
ã®ã¬ã©ã¹ç®¡ã«å å¡«ããŠã¢ãã€ããã€ã¯ããã
ã°ã©ãã€ãŒçšã«ã©ã ãšãããåèšïŒâïŒã®æ
äœã§èª¿è£œããæè¡æž ïŒæäœãå«ãïŒæº¶æ¶²ïŒml
ãäžèšã®ã¢ãã€ããã€ã¯ãããã°ã©ãã€çšã«
ã©ã ã«æµå ¥ãã次ãã§ïŒïœïŒçé žå¡©ç·©è¡é£å¡©
氎溶液ïŒ0.85ïŒ PH7.0ïŒããã«ã©ã ã®æº¶åºæ¶²
ã«èçœãæ€åºãããªããªããŸã§æµããã次ã«
50ïœïŒã°ãªã·ã³âå¡©é žç·©è¡æ°Žæº¶æ¶²ïŒPH4.0ïŒ
ãæ·»å ãã0.5Mé£å¡©æ°Žæº¶æ¶²ãã«ã©ã ã«æµã
ãŠæº¶åºããç»åãæ¡åããããããçŽã¡ã«ç
é žæ°ŽçŽ ãããªãŠã ã§äžæ§ã«ããŠçé žå¡©ç·©è¡é£
塩氎溶液ïŒ0.85ïŒ PH7.0ïŒå¯ŸããŠ72æééæ
ãããïŒ24æéæ¯ã«éæå€æ¶²ã亀æãããïŒã
ãããŠã¢ãã€ããã€ã¯ãããã°ã©ãã€ãŒãçš
ããŠç²Ÿè£œããæ£è è «çã«å¯Ÿããæäœæ°Žæº¶æ¶²ã
åŸãã ãã®ããã«ããŠåŸãæäœã¯ãæ°Žã«å¯æº¶ã§ã
ãããããããã¡ã¿ããŒã«ããšã¿ããŒã«ãã¢
ã»ãã³ããã³ãŒã³çã®ææ©æº¶åªã«ã¯äžæº¶ã§ã
ã€ããããã«ãã®æäœã¯æ·»éå³é¢ã®ç¬¬ïŒå³ã«
瀺ããããªèµ€å€ç·åžåãšç¬¬ïŒå³ã«ç€ºã玫å€ç·
åžåã瀺ããååéã¯ãçŽ15äžã§ãdiscé»æ°
æ³³åã§ã¯Rf0ã0.1ã®éã«ååšããç©è³ªã§ãã€
ãã
第ïŒå³ã¯å®æœäŸïŒã«èšèŒã®æé ã§èª¿è£œã粟補ã
ãæäœã«ã€ããŠã®èµ€å€ç·åžåã¹ãã¯ãã«ã瀺ãã
第ïŒå³ã¯åãã玫å€ç·åžåã¹ãã¯ãã«ã瀺ãã
ãæäœã«ã€ããŠã®èµ€å€ç·åžåã¹ãã¯ãã«ã瀺ãã
第ïŒå³ã¯åãã玫å€ç·åžåã¹ãã¯ãã«ã瀺ãã
Claims (1)
- ãç¹èš±è«æ±ã®ç¯å²ã ïŒ è «çæåãçµåãããã¢ãã€ããã€ã¯ããã
ã°ã©ãã€ãŒã«ãã粟補ããŠåŸããããè «çæåã«
察ããåçš®æäœãšãå°ãªããšãïŒåã®ã¢ããåºå
ã¯ã«ã«ããã·ã«åºãæããæè «çæ§ä»£è¬æ®æå€ã
ã¢ããçµåããŠãªãç©è³ªãæå¹æåãšããæè «ç
å€ã ïŒ æäœã¯ãã¶ã«ã³ãŒã180ãäœè€èºçãâ
1210çœè¡ç ãâ388çœè¡ç ããšãŒãªããçãå
ç°èè «ãæ¥æ§ãªã³ãæ§çœè¡ç ã骚é«çåã¯ãã®ä»
ã®äººçããéžæãããè «çã®æåããèªèµ·ããã
å ç«ã°ãããªã³ç»åãè «çæåãçµåãããã¢ã
ã€ããã€ã¯ãããã°ã©ãã€ãŒã«ãã粟補ãããã®
ã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒã®æè «çå€ã ïŒ æè «çæ§ä»£è¬æ®æå€ã¯ã·ã¿ã©ãã³ãïŒâã¢ã¶
ã°ã¢ãã³ãïŒâãã«ãªããŠã©ã·ã«ãã¡ãœãã¬ãã»
ãŒãããã³ã¢ãããããªã³ãããªãŠã ãããªã矀
ããéžæãããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒã®æè «
çå€ã ïŒ æè «çæ§ä»£è¬æ®æå€ã®ã¢ããåºåã¯ã«ã«ãã
ã·ã«åºã¯å°å ¥ããããã®ã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬
ïŒé åã¯ç¬¬ïŒé èšèŒã®æè «çå€ã ïŒ åèšã¢ããåºåã¯ã«ã«ããã·ã«åºã®å°å ¥ã¯ã
äžè¬åŒïŒžïŒCH2ïŒoCOOHïŒåŒäžïŒžã¯Clåã¯Brã瀺
ããïœã¯ïŒãïŒåã¯ïŒã®æŽæ°ã瀺ãïŒãæããå
åç©ãä»ããŠè¡ãããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒ
ã®æè «çå€ã ïŒ åèšç©è³ªã¯ãåçš®æäœïŒåååœãæè «çæ§ä»£
è¬æ®æå€ãïŒä¹è³10ååçµåãããã®ã§ããç¹èš±
è«æ±ã®ç¯å²ç¬¬ïŒé ä¹è³ç¬¬ïŒé ã®ããããã«èšèŒã®
æè «çå€ã ïŒ æ³šå°å€åœ¢æ ã«ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé ä¹è³
第ïŒé èšèŒã®ããããã«èšèŒã®æè «çå€ã
Priority Applications (12)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP14215379A JPS5665829A (en) | 1979-11-02 | 1979-11-02 | Antitumor agent |
US06/103,474 US4315851A (en) | 1978-12-29 | 1979-12-14 | Pharmaceutical composition having antitumor activity |
SE7910483A SE7910483L (sv) | 1978-12-29 | 1979-12-19 | Farmaceutisk komposition med tumorhemmande effekt |
PH23447A PH16902A (en) | 1978-12-29 | 1979-12-20 | Pharmaceutical composition having anti-tumor activity |
FR797932054A FR2445149B1 (fr) | 1978-12-29 | 1979-12-28 | Composition pharmaceutique ayant une activite antitumorale |
GB7944494A GB2038836B (en) | 1978-12-29 | 1979-12-28 | Antitumour substance |
CH11501/79A CH655010A5 (de) | 1978-12-29 | 1979-12-28 | Pharmazeutischer wirkstoff mit antitumoraktivitaet. |
DE2952690A DE2952690C2 (de) | 1978-12-29 | 1979-12-29 | Antitumorsubstanz, Verfahren zu ihrer Herstellung und diese enthaltendes pharmazeutisches Mittel |
IT28464/79A IT1127324B (it) | 1978-12-29 | 1979-12-31 | Composizione farmaceutica avente attivita' antitumorale |
US06/321,486 US4401592A (en) | 1978-12-29 | 1981-11-16 | Pharmaceutical composition having antitumor activity |
SE8406511A SE8406511D0 (sv) | 1978-12-29 | 1984-12-20 | Farmaceutisk komposition med tumorhemmande effekt |
SE8406510A SE8406510L (sv) | 1978-12-29 | 1984-12-20 | Farmaceutisk komposition med tumorhemmande effekt |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP14215379A JPS5665829A (en) | 1979-11-02 | 1979-11-02 | Antitumor agent |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS5665829A JPS5665829A (en) | 1981-06-03 |
JPS6256137B2 true JPS6256137B2 (ja) | 1987-11-24 |
Family
ID=15308586
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP14215379A Granted JPS5665829A (en) | 1978-12-29 | 1979-11-02 | Antitumor agent |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS5665829A (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH01257331A (ja) * | 1987-12-10 | 1989-10-13 | Fuji Electric Co Ltd | ãã€ã¯ãæ³¢æŽåè£ çœ®ãšãã€ã¯ãæ³¢æŽåæ¹æ³ |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0611714B2 (ja) * | 1983-04-08 | 1994-02-16 | å矜ååŠå·¥æ¥æ ªåŒäŒç€Ÿ | ããå ç«ã°ãããªã³çµåæè «çå€ |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS4962194A (ja) * | 1972-05-10 | 1974-06-17 | ||
JPS5161640A (en) * | 1974-09-20 | 1976-05-28 | Searle & Co | Koshuyozaino seizoho |
JPS51144723A (en) * | 1975-05-27 | 1976-12-13 | Yeda Res & Dev | Antiitumor agent |
-
1979
- 1979-11-02 JP JP14215379A patent/JPS5665829A/ja active Granted
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS4962194A (ja) * | 1972-05-10 | 1974-06-17 | ||
JPS5161640A (en) * | 1974-09-20 | 1976-05-28 | Searle & Co | Koshuyozaino seizoho |
JPS51144723A (en) * | 1975-05-27 | 1976-12-13 | Yeda Res & Dev | Antiitumor agent |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH01257331A (ja) * | 1987-12-10 | 1989-10-13 | Fuji Electric Co Ltd | ãã€ã¯ãæ³¢æŽåè£ çœ®ãšãã€ã¯ãæ³¢æŽåæ¹æ³ |
Also Published As
Publication number | Publication date |
---|---|
JPS5665829A (en) | 1981-06-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US4315851A (en) | Pharmaceutical composition having antitumor activity | |
US4925662A (en) | Anti-tumor substance and process for producing the same | |
US4046722A (en) | Immunological materials | |
US4093607A (en) | Immunologic chemotherapeutic agents comprising antigen binding dimers covalently bound to drugs | |
US4376765A (en) | Medicaments, their preparation and compositions containing same | |
JPH02256700A (ja) | 现èæ¯æ§è¬ç©ã³ã³ãžã¥ã²ãŒã | |
HU189469B (en) | Process for preparing antitumour immunoglobulin-derivatives with antibody effect, comprising ricin a subordinate unit | |
EP0620011A1 (en) | Antibody-drug conjugates | |
JPH02131499A (ja) | 现èæ¯æ§è¬ç©ã³ã³ãžã¥ã²ãŒãããã³ãã®è£œæ³ | |
JPH02191300A (ja) | æäœâè¬ç©æ±åäœ | |
JPH07223969A (ja) | ããªãšãŒãã«çµåäœã®è£œé æ³ | |
US5672688A (en) | Immunoglobulin Fc fragment bound to an alkylating, antibiotic, or antimetabolic antitum or substance | |
JPS6254086B2 (ja) | ||
JPS6256137B2 (ja) | ||
JPS6256136B2 (ja) | ||
JPH08509698A (ja) | èçœè³ªè€åäœããããå«ãçµæç©åã³ãã®å»è¬ãšããŠã®çšé | |
JPH0651643B2 (ja) | 殺现èæ§ä¿®é£Ÿå ç«ã°ãããªã³åã³ãã®è£œé æ¹æ³ | |
NZ225372A (en) | Immunotoxin composition comprising purified ricin-a-chain species | |
CA2121990A1 (en) | Antibody-drug conjugates | |
WO2023232144A1 (zh) | 寡ç³è¿æ¥åïŒå å«å¯¡ç³è¿æ¥åçè¿æ¥å-èŽèœœç©åç³éŸéå¡çæäœå¶èè¯ç©ïŒå ¶å¶å€æ¹æ³åçšé | |
EP0539584A1 (en) | Reagent for diagnosing mycoplasma pneumoniae | |
AU626087B2 (en) | Improved immunotoxin therapies utilizing purified ricin a-chain species | |
JPS62142124A (ja) | ããå ç«ã°ãããªã³çµåæè «çå€ | |
JPS62142123A (ja) | ããå ç«ã°ãããªã³çµåæè «çå€ | |
JPS62283101A (ja) | å¶çäœçšç©è³ªè€åäœã®è£œé æ¹æ³ |