NO154697B - DERIVATIVES OF CANAMYCIN A OR B. - Google Patents
DERIVATIVES OF CANAMYCIN A OR B. Download PDFInfo
- Publication number
- NO154697B NO154697B NO831197A NO831197A NO154697B NO 154697 B NO154697 B NO 154697B NO 831197 A NO831197 A NO 831197A NO 831197 A NO831197 A NO 831197A NO 154697 B NO154697 B NO 154697B
- Authority
- NO
- Norway
- Prior art keywords
- kanamycin
- mmol
- acylation
- acid
- hours
- Prior art date
Links
- 229960000318 kanamycin Drugs 0.000 claims description 137
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical class O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 claims description 105
- 238000002360 preparation method Methods 0.000 claims description 45
- 229960001192 bekanamycin Drugs 0.000 claims description 40
- SKKLOUVUUNMCJE-FQSMHNGLSA-N kanamycin B Chemical compound N[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SKKLOUVUUNMCJE-FQSMHNGLSA-N 0.000 claims description 35
- 229930182824 kanamycin B Natural products 0.000 claims description 30
- 230000000903 blocking effect Effects 0.000 claims description 29
- 239000002253 acid Substances 0.000 claims description 28
- 125000003808 silyl group Chemical group [H][Si]([H])([H])[*] 0.000 claims description 15
- 125000000217 alkyl group Chemical group 0.000 claims description 12
- 229910052739 hydrogen Inorganic materials 0.000 claims description 9
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 claims description 8
- 231100000252 nontoxic Toxicity 0.000 claims description 8
- 230000003000 nontoxic effect Effects 0.000 claims description 8
- 150000003839 salts Chemical class 0.000 claims description 8
- 229910052794 bromium Inorganic materials 0.000 claims description 6
- 229910052801 chlorine Inorganic materials 0.000 claims description 6
- 229910052731 fluorine Inorganic materials 0.000 claims description 6
- 229910052740 iodine Inorganic materials 0.000 claims description 3
- 125000003545 alkoxy group Chemical group 0.000 claims description 2
- 229910007161 Si(CH3)3 Inorganic materials 0.000 claims 1
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 180
- 229930182823 kanamycin A Natural products 0.000 description 99
- 239000000243 solution Substances 0.000 description 73
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 72
- 229910001868 water Inorganic materials 0.000 description 71
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 69
- 238000005917 acylation reaction Methods 0.000 description 69
- LKCWBDHBTVXHDL-RMDFUYIESA-N amikacin Chemical compound O([C@@H]1[C@@H](N)C[C@H]([C@@H]([C@H]1O)O[C@@H]1[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O1)O)NC(=O)[C@@H](O)CCN)[C@H]1O[C@H](CN)[C@@H](O)[C@H](O)[C@H]1O LKCWBDHBTVXHDL-RMDFUYIESA-N 0.000 description 66
- FFUAGWLWBBFQJT-UHFFFAOYSA-N hexamethyldisilazane Chemical compound C[Si](C)(C)N[Si](C)(C)C FFUAGWLWBBFQJT-UHFFFAOYSA-N 0.000 description 66
- 239000000203 mixture Substances 0.000 description 64
- 230000010933 acylation Effects 0.000 description 63
- 229960004821 amikacin Drugs 0.000 description 62
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 42
- 125000001181 organosilyl group Chemical group [SiH3]* 0.000 description 40
- 238000003756 stirring Methods 0.000 description 37
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 30
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 30
- 239000000047 product Substances 0.000 description 30
- 239000003054 catalyst Substances 0.000 description 28
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 25
- 239000000706 filtrate Substances 0.000 description 25
- 239000007787 solid Substances 0.000 description 25
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 24
- FDPIMTJIUBPUKL-UHFFFAOYSA-N pentan-3-one Chemical compound CCC(=O)CC FDPIMTJIUBPUKL-UHFFFAOYSA-N 0.000 description 24
- 238000005406 washing Methods 0.000 description 23
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 21
- 238000006884 silylation reaction Methods 0.000 description 21
- 239000002904 solvent Substances 0.000 description 21
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 20
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 20
- 239000000463 material Substances 0.000 description 20
- 235000016496 Panda oleosa Nutrition 0.000 description 19
- 240000000220 Panda oleosa Species 0.000 description 19
- 238000000034 method Methods 0.000 description 19
- -1 N-protected L-(-)-γ-amino-α-hydroxybutyric acid Chemical class 0.000 description 18
- 238000010992 reflux Methods 0.000 description 17
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 16
- 235000011114 ammonium hydroxide Nutrition 0.000 description 16
- 229930027917 kanamycin Natural products 0.000 description 16
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 15
- SWOHHBNJQHDNFM-WYSMZKEGSA-N (2s)-4-amino-n-[(1s,2r,3r,4s,5r)-5-amino-4-[(2s,3r,4s,5s,6r)-4-amino-3,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-2-[(2r,3r,4s,5s,6r)-6-(aminomethyl)-3,4,5-trihydroxyoxan-2-yl]oxy-3-hydroxycyclohexyl]-2-hydroxybutanamide;sulfuric acid Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.O([C@H]1[C@H](N)C[C@@H]([C@H]([C@@H]1O)O[C@@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CN)O1)O)NC(=O)[C@@H](O)CCN)[C@H]1O[C@H](CO)[C@@H](O)[C@H](N)[C@H]1O SWOHHBNJQHDNFM-WYSMZKEGSA-N 0.000 description 14
- 238000006243 chemical reaction Methods 0.000 description 14
- JHIVVAPYMSGYDF-UHFFFAOYSA-N cyclohexanone Chemical compound O=C1CCCCC1 JHIVVAPYMSGYDF-UHFFFAOYSA-N 0.000 description 14
- 229910052757 nitrogen Inorganic materials 0.000 description 14
- 239000011541 reaction mixture Substances 0.000 description 14
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 13
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Substances C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 13
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 12
- 239000000908 ammonium hydroxide Substances 0.000 description 11
- 239000007864 aqueous solution Substances 0.000 description 11
- 239000003795 chemical substances by application Substances 0.000 description 11
- IJOOHPMOJXWVHK-UHFFFAOYSA-N chlorotrimethylsilane Chemical compound C[Si](C)(C)Cl IJOOHPMOJXWVHK-UHFFFAOYSA-N 0.000 description 11
- 239000007788 liquid Substances 0.000 description 11
- 239000003960 organic solvent Substances 0.000 description 11
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 10
- 238000004458 analytical method Methods 0.000 description 10
- 150000008064 anhydrides Chemical class 0.000 description 10
- 150000001875 compounds Chemical class 0.000 description 10
- 238000010828 elution Methods 0.000 description 10
- NMHMNPHRMNGLLB-UHFFFAOYSA-N phloretic acid Chemical compound OC(=O)CCC1=CC=C(O)C=C1 NMHMNPHRMNGLLB-UHFFFAOYSA-N 0.000 description 10
- 239000007858 starting material Substances 0.000 description 10
- 238000001914 filtration Methods 0.000 description 9
- ADFXKUOMJKEIND-UHFFFAOYSA-N 1,3-dicyclohexylurea Chemical compound C1CCCCC1NC(=O)NC1CCCCC1 ADFXKUOMJKEIND-UHFFFAOYSA-N 0.000 description 8
- WHBMMWSBFZVSSR-UHFFFAOYSA-N 3-hydroxybutyric acid Chemical compound CC(O)CC(O)=O WHBMMWSBFZVSSR-UHFFFAOYSA-N 0.000 description 8
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 8
- 239000002002 slurry Substances 0.000 description 8
- 239000000725 suspension Substances 0.000 description 8
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 7
- 150000002148 esters Chemical class 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 238000004809 thin layer chromatography Methods 0.000 description 7
- ULKOBRDRCYROKY-JTQLQIEISA-N (2s)-2-hydroxy-4-(phenylmethoxycarbonylamino)butanoic acid Chemical compound OC(=O)[C@@H](O)CCNC(=O)OCC1=CC=CC=C1 ULKOBRDRCYROKY-JTQLQIEISA-N 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 239000008346 aqueous phase Substances 0.000 description 6
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 6
- 238000012543 microbiological analysis Methods 0.000 description 6
- WLJVXDMOQOGPHL-UHFFFAOYSA-N phenylacetic acid Chemical compound OC(=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-UHFFFAOYSA-N 0.000 description 6
- 125000000026 trimethylsilyl group Chemical group [H]C([H])([H])[Si]([*])(C([H])([H])[H])C([H])([H])[H] 0.000 description 6
- KYVBNYUBXIEUFW-UHFFFAOYSA-N 1,1,3,3-tetramethylguanidine Chemical compound CN(C)C(=N)N(C)C KYVBNYUBXIEUFW-UHFFFAOYSA-N 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 5
- 150000008065 acid anhydrides Chemical class 0.000 description 5
- 238000009835 boiling Methods 0.000 description 5
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- 239000001257 hydrogen Substances 0.000 description 5
- 239000012299 nitrogen atmosphere Substances 0.000 description 5
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- NTIZESTWPVYFNL-UHFFFAOYSA-N Methyl isobutyl ketone Chemical compound CC(C)CC(C)=O NTIZESTWPVYFNL-UHFFFAOYSA-N 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 125000003277 amino group Chemical group 0.000 description 4
- 239000002585 base Substances 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- DCFKHNIGBAHNSS-UHFFFAOYSA-N chloro(triethyl)silane Chemical compound CC[Si](Cl)(CC)CC DCFKHNIGBAHNSS-UHFFFAOYSA-N 0.000 description 4
- 239000013078 crystal Substances 0.000 description 4
- 238000005828 desilylation reaction Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000012065 filter cake Substances 0.000 description 4
- 229910052736 halogen Inorganic materials 0.000 description 4
- 150000002367 halogens Chemical class 0.000 description 4
- 238000005984 hydrogenation reaction Methods 0.000 description 4
- 238000002955 isolation Methods 0.000 description 4
- IUGYQRQAERSCNH-UHFFFAOYSA-N pivalic acid Chemical compound CC(C)(C)C(O)=O IUGYQRQAERSCNH-UHFFFAOYSA-N 0.000 description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 4
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 3
- CFMZSMGAMPBRBE-UHFFFAOYSA-N 2-hydroxyisoindole-1,3-dione Chemical compound C1=CC=C2C(=O)N(O)C(=O)C2=C1 CFMZSMGAMPBRBE-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 239000005711 Benzoic acid Substances 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 description 3
- 235000010233 benzoic acid Nutrition 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 238000002425 crystallisation Methods 0.000 description 3
- 230000008025 crystallization Effects 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 3
- 238000005342 ion exchange Methods 0.000 description 3
- OOYGSFOGFJDDHP-KMCOLRRFSA-N kanamycin A sulfate Chemical compound OS(O)(=O)=O.O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N OOYGSFOGFJDDHP-KMCOLRRFSA-N 0.000 description 3
- ZUSSTQCWRDLYJA-UHFFFAOYSA-N n-hydroxy-5-norbornene-2,3-dicarboximide Chemical compound C1=CC2CC1C1C2C(=O)N(O)C1=O ZUSSTQCWRDLYJA-UHFFFAOYSA-N 0.000 description 3
- 230000035484 reaction time Effects 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- RMVRSNDYEFQCLF-UHFFFAOYSA-N thiophenol Chemical compound SC1=CC=CC=C1 RMVRSNDYEFQCLF-UHFFFAOYSA-N 0.000 description 3
- PUPZLCDOIYMWBV-UHFFFAOYSA-N (+/-)-1,3-Butanediol Chemical compound CC(O)CCO PUPZLCDOIYMWBV-UHFFFAOYSA-N 0.000 description 2
- MASDFXZJIDNRTR-UHFFFAOYSA-N 1,3-bis(trimethylsilyl)urea Chemical compound C[Si](C)(C)NC(=O)N[Si](C)(C)C MASDFXZJIDNRTR-UHFFFAOYSA-N 0.000 description 2
- QPEJHSFTZVMSJH-UHFFFAOYSA-N 3-amino-5-hydroxybenzoic acid Chemical compound NC1=CC(O)=CC(C(O)=O)=C1 QPEJHSFTZVMSJH-UHFFFAOYSA-N 0.000 description 2
- QUSNBJAOOMFDIB-UHFFFAOYSA-N Ethylamine Chemical compound CCN QUSNBJAOOMFDIB-UHFFFAOYSA-N 0.000 description 2
- 206010019233 Headaches Diseases 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 239000005909 Kieselgur Substances 0.000 description 2
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 description 2
- 230000006181 N-acylation Effects 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 208000010513 Stupor Diseases 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 239000012736 aqueous medium Substances 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethanethiol Chemical compound CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000012458 free base Substances 0.000 description 2
- 150000004820 halides Chemical class 0.000 description 2
- 231100000869 headache Toxicity 0.000 description 2
- 150000002431 hydrogen Chemical class 0.000 description 2
- 238000007327 hydrogenolysis reaction Methods 0.000 description 2
- 239000002085 irritant Substances 0.000 description 2
- 231100000021 irritant Toxicity 0.000 description 2
- GWYFCOCPABKNJV-UHFFFAOYSA-N isovaleric acid Chemical compound CC(C)CC(O)=O GWYFCOCPABKNJV-UHFFFAOYSA-N 0.000 description 2
- 229960000768 kanamycin a sulfate Drugs 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 2
- 239000002808 molecular sieve Substances 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- LWFWUJCJKPUZLV-UHFFFAOYSA-N n-trimethylsilylacetamide Chemical compound CC(=O)N[Si](C)(C)C LWFWUJCJKPUZLV-UHFFFAOYSA-N 0.000 description 2
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- QNGNSVIICDLXHT-UHFFFAOYSA-N para-ethylbenzaldehyde Natural products CCC1=CC=C(C=O)C=C1 QNGNSVIICDLXHT-UHFFFAOYSA-N 0.000 description 2
- XNLICIUVMPYHGG-UHFFFAOYSA-N pentan-2-one Chemical compound CCCC(C)=O XNLICIUVMPYHGG-UHFFFAOYSA-N 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 125000006239 protecting group Chemical group 0.000 description 2
- 238000010926 purge Methods 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 2
- DYHSDKLCOJIUFX-UHFFFAOYSA-N tert-butoxycarbonyl anhydride Chemical compound CC(C)(C)OC(=O)OC(=O)OC(C)(C)C DYHSDKLCOJIUFX-UHFFFAOYSA-N 0.000 description 2
- 125000005931 tert-butyloxycarbonyl group Chemical group [H]C([H])([H])C(OC(*)=O)(C([H])([H])[H])C([H])([H])[H] 0.000 description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- QAEDZJGFFMLHHQ-UHFFFAOYSA-N trifluoroacetic anhydride Chemical compound FC(F)(F)C(=O)OC(=O)C(F)(F)F QAEDZJGFFMLHHQ-UHFFFAOYSA-N 0.000 description 2
- 239000005051 trimethylchlorosilane Substances 0.000 description 2
- 238000010626 work up procedure Methods 0.000 description 2
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- UCKORWKZRPKRQE-UHFFFAOYSA-N bromo(triethyl)silane Chemical compound CC[Si](Br)(CC)CC UCKORWKZRPKRQE-UHFFFAOYSA-N 0.000 description 1
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- PFKFTWBEEFSNDU-UHFFFAOYSA-N carbonyldiimidazole Chemical compound C1=CN=CN1C(=O)N1C=CN=C1 PFKFTWBEEFSNDU-UHFFFAOYSA-N 0.000 description 1
- 238000009903 catalytic hydrogenation reaction Methods 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 239000007810 chemical reaction solvent Substances 0.000 description 1
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- PDNUHAXBKKDGAM-UHFFFAOYSA-N chloro-diethyl-methylsilane Chemical compound CC[Si](C)(Cl)CC PDNUHAXBKKDGAM-UHFFFAOYSA-N 0.000 description 1
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- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 1
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- 238000001514 detection method Methods 0.000 description 1
- 229910000071 diazene Inorganic materials 0.000 description 1
- VLNZUSMTOFYNPS-UHFFFAOYSA-N diethylphosphorylformonitrile Chemical compound CCP(=O)(CC)C#N VLNZUSMTOFYNPS-UHFFFAOYSA-N 0.000 description 1
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- MKRTXPORKIRPDG-UHFFFAOYSA-N diphenylphosphoryl azide Chemical compound C=1C=CC=CC=1P(=O)(N=[N+]=[N-])C1=CC=CC=C1 MKRTXPORKIRPDG-UHFFFAOYSA-N 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
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- ZUKSLMGYYPZZJD-UHFFFAOYSA-N ethenimine Chemical compound C=C=N ZUKSLMGYYPZZJD-UHFFFAOYSA-N 0.000 description 1
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- 229960002064 kanamycin sulfate Drugs 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- TYQCGQRIZGCHNB-JLAZNSOCSA-N l-ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(O)=C(O)C1=O TYQCGQRIZGCHNB-JLAZNSOCSA-N 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 201000003102 mental depression Diseases 0.000 description 1
- 238000006140 methanolysis reaction Methods 0.000 description 1
- 239000005055 methyl trichlorosilane Substances 0.000 description 1
- JLUFWMXJHAVVNN-UHFFFAOYSA-N methyltrichlorosilane Chemical compound C[Si](Cl)(Cl)Cl JLUFWMXJHAVVNN-UHFFFAOYSA-N 0.000 description 1
- 239000003595 mist Substances 0.000 description 1
- LNOPIUAQISRISI-UHFFFAOYSA-N n'-hydroxy-2-propan-2-ylsulfonylethanimidamide Chemical compound CC(C)S(=O)(=O)CC(N)=NO LNOPIUAQISRISI-UHFFFAOYSA-N 0.000 description 1
- 229940078487 nickel acetate tetrahydrate Drugs 0.000 description 1
- OINIXPNQKAZCRL-UHFFFAOYSA-L nickel(2+);diacetate;tetrahydrate Chemical compound O.O.O.O.[Ni+2].CC([O-])=O.CC([O-])=O OINIXPNQKAZCRL-UHFFFAOYSA-L 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
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- 231100000956 nontoxicity Toxicity 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
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- 125000000612 phthaloyl group Chemical group C(C=1C(C(=O)*)=CC=CC1)(=O)* 0.000 description 1
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- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 1
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- 238000005057 refrigeration Methods 0.000 description 1
- FZHAPNGMFPVSLP-UHFFFAOYSA-N silanamine Chemical class [SiH3]N FZHAPNGMFPVSLP-UHFFFAOYSA-N 0.000 description 1
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- 230000009747 swallowing Effects 0.000 description 1
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- BCNZYOJHNLTNEZ-UHFFFAOYSA-N tert-butyldimethylsilyl chloride Chemical compound CC(C)(C)[Si](C)(C)Cl BCNZYOJHNLTNEZ-UHFFFAOYSA-N 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 150000007970 thio esters Chemical class 0.000 description 1
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 1
- 150000003852 triazoles Chemical class 0.000 description 1
- ZOYFEXPFPVDYIS-UHFFFAOYSA-N trichloro(ethyl)silane Chemical compound CC[Si](Cl)(Cl)Cl ZOYFEXPFPVDYIS-UHFFFAOYSA-N 0.000 description 1
- ILWRPSCZWQJDMK-UHFFFAOYSA-N triethylazanium;chloride Chemical compound Cl.CCN(CC)CC ILWRPSCZWQJDMK-UHFFFAOYSA-N 0.000 description 1
- AATYKEFFPLPLST-UHFFFAOYSA-N trimethylsilylurea Chemical compound C[Si](C)(C)NC(N)=O AATYKEFFPLPLST-UHFFFAOYSA-N 0.000 description 1
- 125000002221 trityl group Chemical group [H]C1=C([H])C([H])=C([H])C([H])=C1C([*])(C1=C(C(=C(C(=C1[H])[H])[H])[H])[H])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 1
- 150000003672 ureas Chemical class 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/22—Cyclohexane rings, substituted by nitrogen atoms
- C07H15/222—Cyclohexane rings substituted by at least two nitrogen atoms
- C07H15/226—Cyclohexane rings substituted by at least two nitrogen atoms with at least two saccharide radicals directly attached to the cyclohexane rings
- C07H15/234—Cyclohexane rings substituted by at least two nitrogen atoms with at least two saccharide radicals directly attached to the cyclohexane rings attached to non-adjacent ring carbon atoms of the cyclohexane rings, e.g. kanamycins, tobramycin, nebramycin, gentamicin A2
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Saccharide Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Description
Den foreliggende oppfinnelse vedrører et derivat av kanamycin A eller kanamycin B til anvendelse ved fremstilling av 1-N-|'w-amino-a-hydroksyalkanoyl"J-kanamycin A eller B som har den generelle formel (I) The present invention relates to a derivative of kanamycin A or kanamycin B for use in the production of 1-N-|'w-amino-α-hydroxyalkanoyl"J-kanamycin A or B which has the general formula (I)
hvor R er OH eller NH2 og n er et helt tall fra 0 til 2, eller et ikke-toksisk, farmasøytisk akseptabelt syreaddisjonssalt derav. wherein R is OH or NH 2 and n is an integer from 0 to 2, or a non-toxic, pharmaceutically acceptable acid addition salt thereof.
Kanamycinene er velkjente antibiotika, som f.eks. er beskrevet i Merck Index, 8. utgave, side 597-598. Det er kjent tallrike derivater av kanamycinene. Strukturformlene for kanamycin A og B er angitt nedenfor sammen med det på området anvendte standard nummereringssystem. I det etterfølgende vil de forskjellige derivater av kanamycin, hvor det vil være lett forståelig, bli omtalt som derivater av kanamycin A eller B fremfor ved strukturformelen, slik at behovet for å sammenligne komplekse strukturer for å fastslå forskjeller unngås. The kanamycins are well-known antibiotics, such as e.g. is described in the Merck Index, 8th edition, pages 597-598. Numerous derivatives of the kanamycins are known. The structural formulas for kanamycin A and B are given below together with the standard numbering system used in the field. In the following, the various derivatives of kanamycin, where it will be easily understood, will be referred to as derivatives of kanamycin A or B rather than by the structural formula, so that the need to compare complex structures to determine differences is avoided.
US-patentskrift 3.781.268 vedrører 1- [L-(-)-y-amino-a-hydroksybutyryl] -kanamycin A (amikacin) og B, samt de mono- og dikarbobenzyloksybeskyttende derivater av disse. Når det gjelder lavere og høyere homologer henvises det til US-patentskrifter 3.886.139 og 3.904.597. Forbindelsene fremstilles ved acylering av et 6'-N-beskyttet kanamycin A eller B med et acylerende derivat av en N-beskyttet L-(-)-y-amino-a-hydroksysmørsyre i et vandig medium, etterfulgt av fjerning av den ene N-beskyttende gruppe eller begge N-beskyttende grupper. US patent 3,781,268 relates to 1-[L-(-)-γ-amino-α-hydroxybutyryl]-kanamycin A (amikacin) and B, as well as the mono- and dicarbobenzyloxy-protecting derivatives thereof. Regarding lower and higher homologs, reference is made to US patents 3,886,139 and 3,904,597. The compounds are prepared by acylation of a 6'-N-protected kanamycin A or B with an acylating derivative of an N-protected L-(-)-γ-amino-α-hydroxybutyric acid in an aqueous medium, followed by removal of one N -protecting group or both N-protecting groups.
Fra US-patentskrift 3.974.137 er det kjent en fremgangsmåte til fremstilling av 1-[l-(-)-y-amino-a-hydroksybutyryl]-kanamycin A, som omfatter at man omsetter 6 1 -ka-rbobenzyloksy-kanamycin—A—med— minst 3 mol benzaldehyd, et substituert benzaldehyd eller piv-aldehyd, til dannelse av 6'-N-karbobenzyloksykanamycin A som inneholder Seiffske-basegrupper i 1,3- og 3"-stillingene, acylerer dette tetra-beskyttende kanamycin A derivat med N-hydroksysuccinimidesteren av L-(-)-Y-benzyloksykarbonylamino-a-hydroksysmørsyre og deretter fjerner de beskyttende grupper. From US patent 3,974,137, a method for the production of 1-[1-(-)-γ-amino-α-hydroxybutyryl]-kanamycin A is known, which comprises reacting 6 1 -carbobenzyloxy-kanamycin— A—with— at least 3 moles of benzaldehyde, a substituted benzaldehyde or pivaldehyde, to form 6'-N-carbobenzyloxykanamycin A containing Seiffske base groups in the 1,3 and 3" positions, this acylates tetra-protecting kanamycin A derivative with the N-hydroxysuccinimide ester of L-(-)-Y-benzyloxycarbonylamino-α-hydroxybutyric acid and then remove the protecting groups.
Fra belgisk patentskrift 828.192 er det kjent en fremgangsmåte til fremstilling av 1-[L-(-)-Y-amino-a-hydroksybutyryl ]-kanamycin A ved fremstilling av samme tetra-beskyttende kanamycin A derivat som ifølge US-patentskrift 3.974.137, acylering med N-hydroksy-5-norbornen-2,3-dikarboksimidesteren av L-(-)-y-benzy1-oksykarbonylamino-a-hydroksysmørsyre og etterfølgende fjerning av de beskyttede grupper. From Belgian patent document 828,192, a method for the production of 1-[L-(-)-Y-amino-α-hydroxybutyryl]-kanamycin A is known by producing the same tetra-protective kanamycin A derivative as according to US patent document 3,974,137 , acylation with the N-hydroxy-5-norbornene-2,3-dicarboximide ester of L-(-)-γ-benzy1-oxycarbonylamino-α-hydroxybutyric acid and subsequent removal of the protected groups.
Anvendelsen av et polysilylert kanamycin A eller B som utgangsmateriale ved fremstilling av forbindelser med formelen (I) gir stor løselighet i det organiske løsningsmiddelssystem, noe som muliggjør reaksjon ved høye konsentrasjoner. Selv om reaksjonen vanligvis utføres i løsninger som inneholder 10-20% polysilylert kanamycin-utgangsmateriale, er meget gode resultater blitt oppnådd ved konsentrasjoner på ca. 50 g/100 ml løs-ningsmiddel . The use of a polysilylated kanamycin A or B as starting material in the preparation of compounds of the formula (I) provides high solubility in the organic solvent system, which enables reaction at high concentrations. Although the reaction is usually carried out in solutions containing 10-20% polysilylated kanamycin starting material, very good results have been obtained at concentrations of approx. 50 g/100 ml solvent.
Fremstillingen gir en blanding av acylerte produkter. Det ønskete 1-N-acylerte produkt fraskilles fra de andre produkter ved krom-atografi, og dersom det er ønskelig kan biproduktene hydrolyseres til utgangskanamycinen med henblikk på resirkulering. Det har tidligere vist seg at eventuelt 3"-N-acylert materiale som ble dannet forå rsaket et tap på omtrent en tilsvarende mengde av The preparation gives a mixture of acylated products. The desired 1-N-acylated product is separated from the other products by chromatography, and if desired, the by-products can be hydrolyzed to the starting kanamycin with a view to recycling. It has previously been shown that any 3"-N-acylated material that was formed caused a loss of approximately a corresponding amount of
det ønskete 1-N-acylerte produkt på grunn av den store vanske-lighet med å fraskille sistnevnte fra førstnevnte. Et særlig attraktivt trekk ved fremgangsmåten hvor derivatene ifølge oppfinnelsen anvendes er den meget lave mengde uønsket 3"-N-acylert produkt som dannes (typisk påvises det intet). the desired 1-N-acylated product because of the great difficulty in separating the latter from the former. A particularly attractive feature of the process where the derivatives according to the invention are used is the very low amount of unwanted 3"-N-acylated product that is formed (typically none is detected).
Ved fremstilling av 1- [ L - ( - ) -y-atnino-o - hy droksybutyry 1 ] - kanamycin A, amikacin, ved forskjellige fremgangsmåter, dannes det typisk også det 3"-N-acylerte produkt (BB-K11), det 3-N-acylerte produkt (BB-K29), det 6'-N-acylerte produkt (BB-K6) og polyacylert materiale sammen med ureagert kanamycin A. Ved kom-mersiell fremstilling av amikacin ved acylering av 6'-N-karbobenzyloksy kanamycin A i et vandig medium etterfulgt av fjerning av den beskyttende gruppe, viste det seg således at ca. 10% av det ønskete amikacin (2,5 kg i en sats på 25 kg) vanligvis gikk tapt på grunn av nærværet av BB-Kll som koprodukt. Når amikacin fremstilles ved fremgangsmåten hvor derivatene ifølge oppfinnelsen anvendes, påvises BB-Kll typisk ikke i reaksjonsblandingen. In the production of 1- [ L - ( - ) -y-atnino-o - hydroxybutyry 1 ] - kanamycin A, amikacin, by various methods, the 3"-N-acylated product (BB-K11) is also typically formed, the 3-N-acylated product (BB-K29), the 6'-N-acylated product (BB-K6) and polyacylated material together with unreacted kanamycin A. In the commercial production of amikacin by acylation of 6'-N- carbobenzyloxy kanamycin A in an aqueous medium followed by removal of the protecting group, it was thus found that about 10% of the desired amikacin (2.5 kg in a 25 kg batch) was usually lost due to the presence of BB- Kll as a coproduct When amikacin is prepared by the method in which the derivatives according to the invention are used, BB-Kll is typically not detected in the reaction mixture.
Denne fremgangsmåte omfatter at man This method includes that one
acylerer polysilylert kanamycin A eller B eller polysilylert kanamycin A eller B som inneholder en blokkerende gruppe som er forskjellig fra silyl, på 6'-aminogruppen, med et acylerende derivat av syren med den generelle formel (II): acylates polysilylated kanamycin A or B or polysilylated kanamycin A or B containing a blocking group other than silyl, on the 6'-amino group, with an acylating derivative of the acid of the general formula (II):
hvor n er et helt tall fra 0 til 2 og B er en aminoblokkerende gruppe, i et stort sett vannfritt organisk løsningsmiddel, og at man deretter fjerner alle blokkerende grupper. where n is an integer from 0 to 2 and B is an amino blocking group, in a largely anhydrous organic solvent, and that all blocking groups are then removed.
De blokkerende grupper som kan anvendes til beskyttelse The blocking groups that can be used for protection
av 6'-aminogruppen på kanamycinet og aminogruppen på den acylerende syre (gruppe B i formel II) er konvensjonelle blokkerende grupper for beskyttelse av primære aminogrupper, og de er vel-kjent for fagfolk. Egnete blokkerende grupper omfatter alkoksy-karbonylgrupper, såsom tert-butoksykarbonyl og tert-amyloksy-karbonyl, aralkoksykarbonylgrupper som benzyloksykarbony1, cyklo-alkyloksykarbonylgrupper som cykloheksyloksykarbony1, halogen-alkoksykarbonylgrupper som trikloretoksykarbonyl, acylgrupper som ftaloyl og o-nitrofenoksyacetyl, samt andre kjente blokkerende grupper, såsom o-nitrofenyltiogruppen, tritylgruppen, etc. of the 6'-amino group of the kanamycin and the amino group of the acylating acid (group B in formula II) are conventional blocking groups for protection of primary amino groups and are well known to those skilled in the art. Suitable blocking groups include alkyloxycarbonyl groups, such as tert-butoxycarbonyl and tert-amyloxycarbonyl, aralkyloxycarbonyl groups such as benzyloxycarbonyl, cycloalkyloxycarbonyl groups such as cyclohexyloxycarbonyl, halogeno-alkoxycarbonyl groups such as trichloroethoxycarbonyl, acyl groups such as phthaloyl and o-nitrophenoxyacetyl, as well as other known blocking groups, such as the o-nitrophenylthio group, the trityl group, etc.
Den acylerende syre med formelen (II) kan være i den (+)-eller (-)-isomere form eller en blanding av de to isomerer (d,l-formen), slik at det fremstilles den tilsvarende forbindelse med formelen (I), hvor 1-N-[u-amino-a-hydroksyalkanoyl]-gruppen er i sin (+)- [eller (R)]-form eller sin (-)- [eller (S)]-form, eller en blanding av disse. The acylating acid with the formula (II) can be in the (+)- or (-)-isomeric form or a mixture of the two isomers (d,l-form), so that the corresponding compound with the formula (I) is produced , where the 1-N-[u-amino-α-hydroxyalkanoyl] group is in its (+)- [or (R)] form or its (-)- [or (S)] form, or a mixture of these.
Derivatet ifølge oppfinnelsen er kjennetegnet ved at det har den generelle formel (!')• hvor Z er -0- eller -NH-, R' or H eller en silylgruppe med formelen Si (lavere alkyl)3, minst to og høyst ti R'-grupper er Si (lavere alkyl)^/ og R<1>' er R' eller en annen blokkerende gruppe enn en silylgruppe, og valgt blondt grupper med formlene The derivative according to the invention is characterized by having the general formula (!')• where Z is -0- or -NH-, R' or H or a silyl group with the formula Si (lower alkyl)3, at least two and at most ten R '-groups are Si (lower alkyl)^/ and R<1>' is R' or a blocking group other than a silyl group, and selected blonde groups of the formulas
hvor R 1 og R 2 er like eller forskjellige, og hver er H, F* Cl, Br, NO^ 0H~, —lavere alkyl eller lavere alkoksy, X er Cl, Br, F. eller I, og Y er H, Cl, Br, F eller I. wherein R 1 and R 2 are the same or different and each is H, F* Cl, Br, NO^ 0H~, —lower alkyl or lower alkoxy, X is Cl, Br, F. or I, and Y is H, Cl, Br, F or I.
Det acylerende derivat av syren med formelen (II) er fortrinnsvis en aktiv ester, fortrinnsvis dens aktive ester med N-hydroksyftalimid. Det acylerende derivat av syren med formelen (II) kan også være et blandet syreanhydrid, fortrinnsvis dens blandete syreanhydrid med pivalinsyre, benzoesyre, isobutylkarbonsyre eller benzylkarbonsyre. The acylating derivative of the acid of formula (II) is preferably an active ester, preferably its active ester with N-hydroxyphthalimide. The acylating derivative of the acid of the formula (II) can also be a mixed acid anhydride, preferably its mixed acid anhydride with pivalic acid, benzoic acid, isobutylcarboxylic acid or benzylcarboxylic acid.
Ved fremstilling av 1-N-|-L-(-)-v-amino-^-hydroksybutyryl|-kanamycin A eller et ikke-toksisk, farmasøytisk akseptabelt syreaddisjonssalt av denne omfatter fremgangsmåten at man acylerer polysilylert kanamycin A med et blandet syreanhydrid av L-( - )-\'-benzyloksykarbonylamino-e*-hydroksysmørsyre (fortrinnsvis dens blandete syreanhydrid med pivalinsyre, benzoesyre, isobutyrlkarbonsyre eller benzylkarbonsyre) i et stort sett vannfritt organisk løsningsmiddel, 'og at man deretter fjerner alle blokkerende grupper. In the preparation of 1-N-|-L-(-)-v-amino-^-hydroxybutyryl|-kanamycin A or a non-toxic, pharmaceutically acceptable acid addition salt thereof, the method comprises acylating polysilylated kanamycin A with a mixed acid anhydride of L-( - )-\'-benzyloxycarbonylamino-e*-hydroxybutyric acid (preferably its mixed anhydride with pivalic acid, benzoic acid, isobutyrylcarboxylic acid or benzylcarboxylic acid) in a largely anhydrous organic solvent, and that all blocking groups are then removed.
Ved fremstilling av 1-N-[L-(-)-Y-amino-a-hydroksybutyry1]-kanamycin A eller et ikke-toksisk, farmasøytisk akseptabelt syreaddisjonssalt av denne omfatter fremgangsmåten at man acylerer polysilylert kanamycin A som inneholder en karbobenzyloksygruppe på 6'-aminogruppen med et blandet syreanhydrid av L-(-)-y-benzyloksykarbonylamino-a-hydroksysmørsyre (fortrinnsvis dens blandete syreanhydrid med pivalinsyre, benzoesyre, isobutylkarbonsyre eller benzylkarbonsyre) i et stort sett vannfritt organisk løsningsmiddel, og at man deretter fjerner alle blokkerende grupper. In the preparation of 1-N-[L-(-)-Y-amino-α-hydroxybutyryl]-kanamycin A or a non-toxic, pharmaceutically acceptable acid addition salt thereof, the method comprises acylating polysilylated kanamycin A which contains a carbobenzyloxy group of 6 the '-amino group with a mixed anhydride of L-(-)-γ-benzyloxycarbonylamino-α-hydroxybutyric acid (preferably its mixed anhydride with pivalic acid, benzoic acid, isobutylcarboxylic acid or benzylcarboxylic acid) in a largely anhydrous organic solvent, and that all blocking groups.
Ved fremstilling av 1-N-[L-(-)-Y~amino-a-hydroksybutyry1]-kanamycin A eller et ikke-toksisk, farmasøytisk akseptabelt syre-addis j onssalt av denne omfatter fremgangsmåten-at -man—acylerer - polysilylert kanamycin A med en aktiv ester av L-(-)-Y-benzyloksykarbony lamino-a-hydroksy smørsyre (fortrinnsvis dens aktive ester med N-hydroksysuccinimid, N-hydroksy-5-norbornen-2,3-dikarboks-imid eller N-hydroksyftalimid) i et stort sett vannfritt organisk løsningsmiddel, og at man deretter fjerner alle blokkerende grupper. In the preparation of 1-N-[L-(-)-Y~amino-α-hydroxybutyryl]-kanamycin A or a non-toxic, pharmaceutically acceptable acid addition salt thereof, the method comprises kanamycin A with an active ester of L-(-)-Y-benzyloxycarbonyl lamino-α-hydroxy butyric acid (preferably its active ester with N-hydroxysuccinimide, N-hydroxy-5-norbornene-2,3-dicarboximide or N- hydroxyphthalimide) in a largely anhydrous organic solvent, and that all blocking groups are then removed.
Ved fremstilling av 1-N-[L-(-)-Y-amino-a-hydroksybutyry1]-kanamycin A eller et ikke-toksisk, farmasøytisk akseptabelt syre-addis jonssalt av denne omfatter fremgangsmåten at man acylerer polysilylert kanamycin A som inneholder en karbobenzyloksygruppe på 6<1->aminogruppen, med en aktiv ester av L-(-)-Y-benzyloksykarbony lamino-a-hydroksysmørsyre (fortrinnsvis dens aktive ester med N-hydroksysuccinimid, N-hydroksy-5-norbornen-2,3-dikarboks-imid eller N-hydroksyftalimid) i et stort sett vannfritt organisk løsningsmiddel og at man deretter fjerner alle blokkerende grupper. In the preparation of 1-N-[L-(-)-Y-amino-α-hydroxybutyryl]-kanamycin A or a non-toxic, pharmaceutically acceptable acid addition salt thereof, the method comprises acylating polysilylated kanamycin A which contains a carbobenzyloxy group on the 6<1->amino group, with an active ester of L-(-)-Y-benzyloxycarbonyl lamino-α-hydroxybutyric acid (preferably its active ester with N-hydroxysuccinimide, N-hydroxy-5-norbornene-2,3- dicarboximide or N-hydroxyphthalimide) in a largely anhydrous organic solvent and that all blocking groups are then removed.
I en foretrukket utførelsesform er derivatet polysilylert kanamycin A eller B (fortrinnsvis polysilylert kanamycin A) som inneholder et gjennomsnittlig antall silylgrupper (fortrinnsvis trimetylsilylgrupper) pr. molekyl på fra 4 til 8. I en annen foretrukket utførelsesform er derivatet polysilylert kanamycin A eller B (fortrinnsvis polysilylert kanamycin A) som inneholder en annen blokkerende gruppe enn silyl på 6'-aminogruppen og inneholder et gjennomsnittlig antall silylgrupper (fortrinnsvis trimetylsilylgrupper) pr molekyl på f.r_a_3_ til 7. In a preferred embodiment, the derivative is polysilylated kanamycin A or B (preferably polysilylated kanamycin A) which contains an average number of silyl groups (preferably trimethylsilyl groups) per molecule of from 4 to 8. In another preferred embodiment, the derivative is polysilylated kanamycin A or B (preferably polysilylated kanamycin A) which contains a blocking group other than silyl on the 6'-amino group and contains an average number of silyl groups (preferably trimethylsilyl groups) per molecule on f.r_a_3_ to 7.
Betegnelsen "ikke-toksisk, farmasøytisk akseptabelt syre-addis jonssalt" av en. forbindelse med formelen (I) betyr her et The term "non-toxic, pharmaceutically acceptable acid addition salt" of a. connection with the formula (I) here means a
nono-, di-, tri- eller tetrasalt dannet ved reaksjon mellom ett molekyl av en forbindelse med formelen (I) og 1-4 ekvivalenter av en ikke-toksisk, farmasøytisk akseptabel syre. Disse syrer omfatter eddiksyre, saltsyre, svovelsyre, maleinsyre, fosfor-syre, salpetersyre, hydrobromsyre, askorbinsyre, eplesyre og sitronsyre, samt de andre syrer som vanligvis anvendes til fremstilling av salter av aminoholdige legemidler. nono-, di-, tri- or tetrasalt formed by reaction between one molecule of a compound of formula (I) and 1-4 equivalents of a non-toxic, pharmaceutically acceptable acid. These acids include acetic acid, hydrochloric acid, sulfuric acid, maleic acid, phosphoric acid, nitric acid, hydrobromic acid, ascorbic acid, malic acid and citric acid, as well as the other acids which are usually used for the production of salts of amino-containing drugs.
Acylering av det polysilylerte kanamycin A eller B (med eller uten en blokkerende gruppe som er forskjellig fra silyl på 6'-aminogruppen) kan vanligvis utføres i et organisk løsnings-middel som utgangsmaterialet har tilstrekkelig løselighet i. Disse materialer er godt løselige i de fleste vanlige organiske løs-ningsmidler. Egnete løsningsmidler er f.eks. aceton, dietylketon, metyl-n-propylketon, metylisobutylketon, metyletylketon, acetonitril, glyme, diglyme, dioksan, toluen, tetrahydrofuran, cykloheksanon, metylenklorid, kloroform, karbontetraklorid og blan-dinger av aceton/butanol eller dietylketon/butanol. Valget av løsningsmiddel avhenger av det spesielle utgangsmateriale som anvendes. Ketoner er vanligvis de foretrukne løsningsmidler. Acylation of the polysilylated kanamycin A or B (with or without a blocking group other than silyl on the 6'-amino group) can usually be carried out in an organic solvent in which the starting material has sufficient solubility. These materials are well soluble in most common organic solvents. Suitable solvents are e.g. acetone, diethyl ketone, methyl-n-propyl ketone, methyl isobutyl ketone, methyl ethyl ketone, acetonitrile, glyme, diglyme, dioxane, toluene, tetrahydrofuran, cyclohexanone, methylene chloride, chloroform, carbon tetrachloride and mixtures of acetone/butanol or diethyl ketone/butanol. The choice of solvent depends on the particular starting material used. Ketones are usually the preferred solvents.
Det mest fordelaktige løsningsmiddel for den spesielle kombinasjon av reaksjonsdeltakere som anvendes kan lettvint bestemmes ved rutineforsøk. The most advantageous solvent for the particular combination of reaction participants used can easily be determined by routine testing.
Egnete silyleringsmidler for anvendelse ved fremstilling av de polysilylerte kanamyciner omfatter silyleringsmidlene med de generelle formler: Suitable silylating agents for use in the preparation of the polysilylated kanamycins include the silylating agents of the general formulas:
5 6 1 hvor R , R og R er hydrogen, halogen, (lavere)alkyl, halogen-(lavere)alkyl eller fenyl, idet minst én av gruppene R 5 , R 6 og R 7 ikke er halogen eller hydrogen, R 4 er (lavere)alkyl, m er et helt tall på 1 eller 2 og X er halogen eller hvor R 8 er hydrogen eller (lavere)alkyl og R <9>er hydrogen, (lavere)alkyl eller 5 6 1 where R , R and R are hydrogen, halogen, (lower) alkyl, halogen (lower) alkyl or phenyl, with at least one of the groups R 5 , R 6 and R 7 not being halogen or hydrogen, R 4 being (lower) alkyl, m is an integer of 1 or 2 and X is halogen or where R 8 is hydrogen or (lower) alkyl and R<9> is hydrogen, (lower) alkyl or
hvor R^, R^ og R7 har den ovenfor angitte betydning. where R^, R^ and R7 have the meaning indicated above.
Spesifikke silylforbindelser med formlene IV og V er: trimetylklorsilan, heksametyldisilazan, trietylklorsilan, metyl-triklorsilan, dimetyldiklorsilan, trietylbromsilan, tri-n-pro-pylklorsilan, metyldietylklorsilan, dimetyletylklorsilan, di-metyl-t-butylklorsilan, fenyldimetylbromsilan, benzylmetylety1-klorsilan, fenyletylmetylklorsilan, trifenylklorsilan, trifenyl-fluorsilan, tri-o-tolylklorsilan, tri- p- dimety laminof enyJLklor-silan, N-etyltrietylsilylamin, heksaetyldisilazan, trifenylsilyl-amin, tri-n-propylsilylamin, tetraetyldimetyldisilazan, heksa-fenyldisilazan, heksa-p-tolyldisilazan, etc. Også heksaalkyl-cyklotrisilazaner og okta-alkylcyklotetrasilazaner er anvend-bare. Andre egnete silyleringsmidler er silylamider (såsom trialkylsilylacetamider og bis-trialkylsilylacetamider), silylurea såsom trimetylsilylurea samt silylureider. Også trimetylsily1-imidazol kan anvendes. Specific silyl compounds of formulas IV and V are: trimethylchlorosilane, hexamethyldisilazane, triethylchlorosilane, methyltrichlorosilane, dimethyldichlorosilane, triethylbromosilane, tri-n-propylchlorosilane, methyldiethylchlorosilane, dimethylethylchlorosilane, dimethyl-t-butylchlorosilane, phenyldimethylbromosilane, benzylmethylethylchlorosilane, phenylethylchloromethylchlorosilane. . , etc. Also hexaalkyl-cyclotrisilazanes and octa-alkylcyclotetrasilazanes are usable. Other suitable silylating agents are silylamides (such as trialkylsilylacetamides and bis-trialkylsilylacetamides), silylureas such as trimethylsilylurea and silylureides. Trimethylsily1-imidazole can also be used.
En foretrukket silylgruppe er trimetylsilylgruppen, og foretrukne silyleringsmidler for innføring av trimetylsilylgruppen er heksametyldisilazan, bis-(trimetylsilyl)-acetamid samt trimetylsilylacetamid. Heksametyldisilazan er det mest foretrukne. A preferred silyl group is the trimethylsilyl group, and preferred silylating agents for introducing the trimethylsilyl group are hexamethyldisilazane, bis-(trimethylsilyl)-acetamide and trimethylsilylacetamide. Hexamethyldisilazane is most preferred.
Når det anvendes polysilylert kanamycin A eller B som inneholder en annen blokkerende gruppe enn silyl på 6<1->aminogruppen, kan dette fremstilles enten ved polysilylering av det ønskete 6'-N-blokkerte kanamycin A eller B, eller ved innføring av den ønskete 6'-blokkerende gruppe i polysilylert kanamycin A eller B. When polysilylated kanamycin A or B containing a blocking group other than silyl on the 6<1->amino group is used, this can be produced either by polysilylation of the desired 6'-N-blocked kanamycin A or B, or by introducing the desired 6' blocking group in polysilylated kanamycin A or B.
Fremgangsmåter for innføring av silylgrupper i organiske forbindelser, inklusive visse aminoglykosider, er kjent på området. De polysilylerte kanamyciner (med eller uten en annen blokkerende gruppe enn silyl på 6'-aminogruppen) kan fremstilles ved i og for seg kjente fremgangsmåter eller slik som beskrevet i denne beskrivelse. Methods for introducing silyl groups into organic compounds, including certain aminoglycosides, are known in the field. The polysilylated kanamycins (with or without a blocking group other than silyl on the 6'-amino group) can be prepared by methods known per se or as described in this description.
Betegnelsen polysilylert kanamycin A eller B refererer slik den er anvendt her til kanamycin A eller B som inneholder fra 2 til 10 silylgrupper i molekylet. Betegnelsen polysilylert kanamycin A eller B omfatter således ikke persilylert kanamycin A eller B, som ville inneholde 11 silylgrupper i molekylet. The term polysilylated kanamycin A or B as used herein refers to kanamycin A or B containing from 2 to 10 silyl groups in the molecule. The term polysilylated kanamycin A or B thus does not include persilylated kanamycin A or B, which would contain 11 silyl groups in the molecule.
Det nøyaktige antall silylgrupper som er til stede i de polysilylerte kanamycin-utgangsmaterialer (med eller uten en blokkerende gruppe som er forskjellig fra silyl på 6'-aminogruppen) eller deres stilling er ikke kjent. Det har vist seg at både undersilylering og oversilylering minsker utbyttet av det ønskete produkt og øker utbyttet av andre produkter. I tilfelle kraftig under- og oversilylering kan det bli dannet litt eller over hode ikke noe av det ønskete produkt. Den silyleringsgrad som vil gi det største utbytte av ønsket produkt vil avhenge av de spesielle reaksjonsdeltakere som anvendes i acyleringstrinnet. Den mest fordelaktige silyleringsgrad under anvendelse av en vilkårlig kombinasjon av reaksjonsdeltakere kan lettvint fastslåes ved rutineforsøk. The exact number of silyl groups present in the polysilylated kanamycin starting materials (with or without a blocking group other than silyl on the 6'-amino group) or their position is not known. It has been shown that both undersilylation and oversilylation reduce the yield of the desired product and increase the yield of other products. In case of strong under- and over-silylation, little or none of the desired product may be formed. The degree of silylation which will give the greatest yield of the desired product will depend on the particular reaction participants used in the acylation step. The most advantageous degree of silylation using an arbitrary combination of reaction participants can easily be determined by routine experiments.
Ved fremstilling av 1-N- [ h-(-)-y-amino-a-hydroksybutyry1] - kanamycin A ved acylering av polysilylert kanamycin A med N-hydroksysuccinimidesteren av L-(-)-y-benzyloksykarbonylamino-a-hydroksysmørsyre i acetonløsning har det vist seg at gode utbytter av det ønskete produkt oppnås ved anvendelse av polysilylert kanamycin A, som er blitt fremstilt ved omsetning av fra 4 til In the preparation of 1-N-[h-(-)-y-amino-α-hydroxybutyryl]-kanamycin A by acylation of polysilylated kanamycin A with the N-hydroxysuccinimide ester of L-(-)-y-benzyloxycarbonylamino-α-hydroxybutyric acid in acetone solution, it has been shown that good yields of the desired product are obtained by using polysilylated kanamycin A, which has been prepared by reacting from 4 to
5,5 mol heksametyldisilazan pr. mol kanamycin A. Større eller mTfrdr"é~~mÆngder héksame~tyldisilazan kan anvendes, men utbyttet 5.5 mol hexamethyldisilazane per moles of kanamycin A. Larger or lower amounts of hexamethyldisilazane can be used, but the yield
av det ønskete produkt i det etterfølgende acyleringstrinn minskes betraktelig. Ved den spesielle fremgangsmåte som er anført ovenfor, foretrekkes det å anvende fra 4,5 til 5,0 mol heksametyldisilazan pr. mol kanamycin med henblikk på oppnåelse av maksi-malt produktutbytte i acyleringstrinnet. of the desired product in the subsequent acylation step is considerably reduced. In the particular method stated above, it is preferred to use from 4.5 to 5.0 mol of hexamethyldisilazane per mol of kanamycin with a view to achieving maximum product yield in the acylation step.
Det vil forståes at hvert mol heksametyldisilazan er i stand til å innføre to ekvivalenter av trimetylsilylgruppen i kanamycin A eller B. Både kanamycin A og B har ialt 11 stillinger (NH2- og OH-grupper) som kan silyleres, mens kanamycin A og B som inneholder en blokkerende gruppe som er forskjellig fra silyl på 6'-aminogruppen, har totalt 10 slike stillinger. 5,5 mol heksametyldisilazan pr. mol kanamycin A eller B kunne således teoretisk silylere alle OH- og N^-gruppene på kanamycinet fullstendig, mens 5,0 mol heksametyldisilazan fullstendig kunne silylere ett mol kanamycin A eller B som inneholder en blokkerende gruppe forskjellig fra silyl på 6'-aminogruppen. Det menes imidlertid at en så vidtgående silylering ikke finner sted med disse molare forhold innenfor rimelige reaksjonstidsrom, selv om høyere silyleringsgrader oppnås i løpet av et gitt reaksjonstidsrom når det tilsettes en silyleringskatalysator. It will be understood that each mole of hexamethyldisilazane is capable of introducing two equivalents of the trimethylsilyl group into kanamycin A or B. Both kanamycin A and B have a total of 11 positions (NH2 and OH groups) which can be silylated, while kanamycin A and B which contains a blocking group different from silyl on the 6'-amino group, has a total of 10 such positions. 5.5 mol hexamethyldisilazane per mol of kanamycin A or B could thus theoretically completely silylate all OH and N^ groups on the kanamycin, while 5.0 mol of hexamethyldisilazane could completely silylate one mol of kanamycin A or B containing a blocking group different from silyl on the 6'-amino group. However, it is believed that such extensive silylation does not take place with these molar ratios within reasonable reaction times, even if higher degrees of silylation are achieved during a given reaction time when a silylation catalyst is added.
Silyleringskatalysatorer akselererer i høy grad silyler-ingshastigheten. Egnete silyleringskatalysatorer er velkjente på området og omfatter blant annet aminsulfater, f.eks. kanamycin-sulfat, sulfaminsyre, imidazol samt trimetylklorsilan. Silyleringskatalysatorer fremmer vanligvis en høyere silyleringsgrad enn det som er nødvendig. Silylation catalysts greatly accelerate the silylation rate. Suitable silylation catalysts are well known in the field and include, among other things, amine sulphates, e.g. kanamycin sulfate, sulfamic acid, imidazole and trimethylchlorosilane. Silylation catalysts usually promote a higher degree of silylation than is necessary.
Oversilylert kanamycin A eller B kan imidlertid anvendes som utgangsmateriale dersom det først behandles med et desilyleringsmiddel for minskning av silyleringsgraden før acyleringsreaksjonen utføres. However, oversilylated kanamycin A or B can be used as starting material if it is first treated with a desilylation agent to reduce the degree of silylation before the acylation reaction is carried out.
Gode utbytter av det ønskete produkt oppnås når man acylerer polysilylert kanamycin A fremstilt under anvendelse av et 5,5:1 molart forhold av heksametyldisilazan til kanamycin A. Når imidlertid kanamycin A silylert med et 7:1 molart forhold av heksametyldisilazan (eller med et 5,5:1 molart forhold under nærvær av en silyleringskatalysator) ble acylert i aceton med N-hydroksysuccinimidesteren av L-(-)-y-benzyloksykarbonylamino-a-hydroksy-smørsyre, ble det oppnådd mindre enn 1% utbytte av det ønskete produkt. Når imidlertid samme "oversilylerte" kanamycin A ble acylert med samme acyleringsmiddel i acetonløsning, som var tilsatt vann (21 mol vann pr. mol kanamycin, 2,5% vann(vekt/volum) som desilyleringsmiddel 1 time før acyleringen, ble det oppnådd et utbytte på ca. 4 0% av det ønskete produkt. Samme resultater oppnås dersom vannet erstattes med metanol eller en annen aktiv hydrogenforbindelse som er i stand til å bevirke desilylering, f.eks. etanol, propanol, butandiol, metylmerkaptan, etylmerkaptan, fenylmerkaptan eller liknende. Good yields of the desired product are obtained when acylating polysilylated kanamycin A prepared using a 5.5:1 molar ratio of hexamethyldisilazane to kanamycin A. However, when kanamycin A silylated with a 7:1 molar ratio of hexamethyldisilazane (or with a 5 .5:1 molar ratio in the presence of a silylation catalyst) was acylated in acetone with the N-hydroxysuccinimide ester of L-(-)-γ-benzyloxycarbonylamino-α-hydroxybutyric acid, less than 1% yield of the desired product was obtained. However, when the same "oversilylated" kanamycin A was acylated with the same acylating agent in acetone solution, to which water had been added (21 mol of water per mol of kanamycin, 2.5% water (w/v) as desilylating agent 1 hour before the acylation, a yield of about 40% of the desired product. The same results are obtained if the water is replaced by methanol or another active hydrogen compound capable of effecting desilylation, e.g. ethanol, propanol, butanediol, methyl mercaptan, ethyl mercaptan, phenyl mercaptan or similar.
Selv om det er vanlig å anvende tørre løsningsmidler når det arbeides med silylerte materialer, har det overraskende vist seg at tilsetning av vann til reaksjonsløsningsmidlet før acylering selv uten "oversilylering" ofte gir like gode utbytter og av og til bedre utbytter av ønsket produkt enn et tørt løsnings-middel. Ved acyleringsreaksjoner gjennomført i aceton i de vanlige konsentrasjoner på 10-20% (vekt/volum) av polysilylert kanamycin A, har det vist seg at meget høye utbytter av 1-N-[ h-(-)-y-amino-a-hydroksybutyryl]-kanamycin A ble oppnådd når det ble tilsatt inntil 28 mol vann pr. mol polysilylert kanamycin A. Ved en konsentrasjon på 20% er 28 mol pr. mol ca. 8% vann. Ved andre kombinasjoner av reaksjonsdeltakere kan sogar mer vann tolereres eller være gunstig. Acyleringsreaksjonen kan utføres i løsnings-midler som inneholder opptil ca. 4 0% vann, selv om man ved så Although it is common to use dry solvents when working with silylated materials, it has surprisingly been found that adding water to the reaction solvent prior to acylation even without "oversilylation" often gives equally good yields and sometimes better yields of the desired product than a dry solvent. In acylation reactions carried out in acetone in the usual concentrations of 10-20% (w/v) of polysilylated kanamycin A, it has been shown that very high yields of 1-N-[ h-(-)-y-amino-a- hydroxybutyryl]-kanamycin A was obtained when up to 28 mol of water per mol polysilylated kanamycin A. At a concentration of 20%, 28 mol per moles approx. 8% water. With other combinations of reaction participants, even more water can be tolerated or be beneficial. The acylation reaction can be carried out in solvents containing up to approx. 4 0% water, even if you know
høye vannkonsentrasjoner må anvende korte acyleringstider for å high water concentrations must use short acylation times to
unngå for sterk desilylering av det polysilylerte kanamycin A avoid too strong desilylation of the polysilylated kanamycin A
eller B utgangsmateriale. Følgelig er det hensikten med betegn- or B starting material. Consequently, it is the purpose of denoting
elsen "stort sett vannfritt organisk løsningsmiddel" slik det er anvendt her å omfatte løsningsmidler som inneholder opptil ca. The term "largely anhydrous organic solvent" as used here includes solvents containing up to approx.
25% vann. Et foretrukket intervall er opptil ca. 20%, et mer foretrukket intervall er opptil 8% vann, og et særlig fore- 25% water. A preferred interval is up to approx. 20%, a more preferred interval is up to 8% water, and a particularly
trukket intervall er opptil ca. 4% vann. drawn interval is up to approx. 4% water.
Som anført ovenfor kan den mest ønskelige silylerings- As stated above, the most desirable silylation
grad for en vilkårlig kombinasjon av acyleringsreaksjonsdeltakere lettvint bestemmes ved rutineforsøk. Det menes at det foretrukne gjennomsnittlige antall silylgrupper i utgangsmaterialet van- degree for an arbitrary combination of acylation reaction participants is easily determined by routine experiments. It is believed that the preferred average number of silyl groups in the starting material
ligvis vil være mellom 4 og 8 for kanamycin A eller B og mellom 3 og 7 for kanamycin A.eller B som inneholder en blokkerende would normally be between 4 and 8 for kanamycin A or B and between 3 and 7 for kanamycin A or B containing a blocking
gruppe forskjellig fra silyl på 6'-aminogruppen, men dette er bare teori og betraktes ikke som en vesentlig del av oppfinnelsen. group other than silyl on the 6'-amino group, but this is only theory and is not considered an essential part of the invention.
Varigheten av og temperaturen under acyleringsreaksjonen The duration and temperature during the acylation reaction
er ikke kritisk. Temperaturer i intervallet fra -30 til 100°C is not critical. Temperatures in the range from -30 to 100°C
kan anvendes ved reaksjonstider i området fra ca. 1 time og can be used for reaction times in the range from approx. 1 hour and
"opptil en dag eller~mer. Det har vist seg at reaksjonen vanligvis forløper godt ved romtemperatur, og av bekvemmelighets grunner og økonomiske grunner foretrekkes det å utføre reaksjonen ved omgivel-sestemperatur. Med henblikk på oppnåelse av maksimale utbytter og selektiv acylering foretrekkes det imidlertid å utføre acyleringen ved fra 0 til 5°C. "up to a day or more. It has been found that the reaction usually proceeds well at room temperature, and for reasons of convenience and economy it is preferred to carry out the reaction at ambient temperature. However, for the purpose of obtaining maximum yields and selective acylation, it is preferred to carry out the acylation at from 0 to 5°C.
Acylering av 1-aminogruppen i det polysilylerte kanamycin Acylation of the 1-amino group in the polysilylated kanamycin
A eller B (med eller uten en blokkerende gruppe som er forskjellig fra silyl på 6<1->aminogruppen) kan utføres med et vilkårlig acyl-eringsderivat av syren med formelen II, som er kjent på området at er egnet til acylering av en primær aminogruppe. Eksempler på egnete acyleringsderivater av den frie syre omfatter de tilsvarende syreanhydrider, blandete anhydrider, f.eks. alkoksymaursyreanhyd-rider, syrehalogenider, syreazider, aktive estre og aktive tio-estre. Den frie syre kan koples med det polysilylerte kanamycin-utgangsmateriale etter at den frie syre først er omsatt med N,N'- A or B (with or without a blocking group other than silyl on the 6<1-> amino group) can be carried out with any acylation derivative of the acid of formula II known in the art to be suitable for acylation of a primary amino group. Examples of suitable acylation derivatives of the free acid include the corresponding acid anhydrides, mixed anhydrides, e.g. alkoxyformic anhydrides, acid halides, acid azides, active esters and active thioesters. The free acid can be coupled with the polysilylated kanamycin starting material after the free acid is first reacted with N,N'-
dimetylkloroforminiumklorid [se britisk patentskrift 1.008.17 0 dimethylchloroforminium chloride [see British patent document 1.008.17 0
og Novak og Weichet, Experientia XXI, 6, 360 (1065)] eller ved anvendelse av et N,N<*->karbonyldiimidazol eller N,N<1->karbonyl-ditriazol (se sydafrikansk patentskrift 63/2684) eller et karbo-diimidreagens [særlig N,N<1->dicykloheksylkarbodiimid, N,N'-di-isopropylkarbodiimid eller N-cykloheksyl-N'-(2-morfolinoetyl)-karbodiimid: se Sheenan og Hess, J.A.C.S., 77 1967 (1955)] , eller av et alkynylaminreagens [se R. Buijle og H.G. Viehe, angew. Chem. International Edition, 3, 582 (1964)] eller av et isoksazoliumsaltreagens [se R.B. Woodward, R.A. Olofson og H. Mayer, J. Amer. Chem. Soc, 83, 1010 (1961)] , eller av et keten-iminreagens [se CL. Stevens og M.E. Munk, J. Amer. Chem. Soc, 80, 4065 (1958)J eller av heksaklorcyklotrifosfatriazin eller heksabromcyklotrifosfatriazin (US-patentskrift 3.651.050) eller av difenylfosforylazid [DDPA, J. Amer. Chem. Soc., 94, 6203-6205 and Novak and Weichet, Experientia XXI, 6, 360 (1065)] or by using an N,N<*->carbonyldiimidazole or N,N<1->carbonyl-ditriazole (see South African patent document 63/2684) or a carb -diimide reagent [especially N,N<1->dicyclohexylcarbodiimide, N,N'-di-isopropylcarbodiimide or N-cyclohexyl-N'-(2-morpholinoethyl)-carbodiimide: see Sheenan and Hess, J.A.C.S., 77 1967 (1955)] , or of an alkynylamine reagent [see R. Buijle and H.G. Viehe, Angew. Chem. International Edition, 3, 582 (1964)] or by an isoxazolium salt reagent [see R.B. Woodward, R.A. Olofson and H. Mayer, J. Amer. Chem. Soc, 83, 1010 (1961)] , or by a ketene-imine reagent [see CL. Stevens and M.E. Munk, J. Amer. Chem. Soc, 80, 4065 (1958)J or of hexachlorocyclotriphosphatetriazine or hexabromocyclotriphosphatetriazine (US Patent 3,651,050) or of diphenylphosphoryl azide [DDPA, J. Amer. Chem. Soc., 94, 6203-6205
(1972)]] eller av dietylfosforylcyanid [DEPC, Tetrahedron Letters (1972)]] or of diethyl phosphoryl cyanide [DEPC, Tetrahedron Letters
nr. 18, sider 1595-1598 (1973)] eller av difenylfosfit [Tetrahedron Letters nr. 49, sider 5047-5050 (1972)]. Et annet derivat som er ekvivalent med syren er et tilsvarende azolid, dvs. et amid av den tilsvarende syre hvis amidnitrogen inngår i en quasiaroma-tisk 5-leddet ring som inneholder minst to nitrogenatomer, dvs. imidazol, pyrazol, triazolene, benzimidazol, benzotriazol og deres substituerte derivater. Som fagfolk vil forstå kan det av og til være Ønskelig eller nødvendig å beskytte hydroksylgruppen på det acylerte derivat av syren med formelen (II), f.eks. når det anvendes acyleringsderivater såsom et syrehalogenid. Beskyttelse av hydroksylgruppen kan utføres på måter som er kjent på området^—f—eks.—ved anvendelse av en karbobenzyloksygruppe, ved acetylering, ved silylering eller liknende. No. 18, pages 1595-1598 (1973)] or of diphenyl phosphite [Tetrahedron Letters No. 49, pages 5047-5050 (1972)]. Another derivative equivalent to the acid is a corresponding azolide, i.e. an amide of the corresponding acid whose amide nitrogen forms part of a quasiaromatic 5-membered ring containing at least two nitrogen atoms, i.e. imidazole, pyrazole, triazoles, benzimidazole, benzotriazole and their substituted derivatives. As those skilled in the art will appreciate, it may occasionally be desirable or necessary to protect the hydroxyl group on the acylated derivative of the acid of formula (II), e.g. when using acylation derivatives such as an acid halide. Protection of the hydroxyl group can be carried out in ways known in the art^—for example—by using a carbobenzyloxy group, by acetylation, by silylation or the like.
Etter avslutning av acyleringsreaksjonen fjernes alle blokkerende grupper på i og for seg kjent måte til dannelse av det ønskete produkt med formelen (I). Silylgruppene kan f.eks. lettvint fjernes ved hydrolyse med vann, fortrinnsvis ved lav pH-verdi. Også den blokkerende gruppe B på acyleringsderivatet av syren med formelen (II) og den blokkerende gruppe på 6'-aminogruppen på det polysilylerte kanamycin-utgangsmateriale (dersom denne er til stede) kan fjernes ved hjelp av kjente metoder. Således kan en tert-butoksykarbonylgruppe fjernes under anvendelse av maursyre, en karbobenzyloksygruppe ved katalytisk hydrogenering, en 2-hydroksy-l-naftkarbonylgruppe ved sur hydrolyse, en triklor-etoksykarbonylgruppe ved behandling med sinkstøv i iseddik, ftaloyl-i 78 timer. Den gule, klare løsning (pH 7,0) ble tynnet med 100 ml vann. Blandingens pH-verdi ble innstilt på 2,8 (3N HCl), og blandingen ble omrørt kraftig ved 23°C i 15 minutter. Den vandige fase ble fraskilt, og den organiske fase ble ekstrahert med 50 After completion of the acylation reaction, all blocking groups are removed in a manner known per se to form the desired product with the formula (I). The silyl groups can e.g. easily removed by hydrolysis with water, preferably at a low pH value. Also, the blocking group B on the acylation derivative of the acid of formula (II) and the blocking group on the 6'-amino group on the polysilylated kanamycin starting material (if present) can be removed by means of known methods. Thus, a tert-butoxycarbonyl group can be removed using formic acid, a carbobenzyloxy group by catalytic hydrogenation, a 2-hydroxy-l-naphthocarbonyl group by acid hydrolysis, a trichloroethoxycarbonyl group by treatment with zinc dust in glacial acetic acid, phthaloyl-for 78 hours. The yellow, clear solution (pH 7.0) was diluted with 100 ml of water. The pH of the mixture was adjusted to 2.8 (3N HCl) and the mixture was stirred vigorously at 23°C for 15 minutes. The aqueous phase was separated, and the organic phase was extracted with 50
ml vann med pH 2,8. De kombinerte vandige fraksjoner ble vasket med 50 ml etylacetat. Løsningen ble anbrakt i en 500 ml Parr-flaske sammen med 5 g 5% palladium-på-karbon katalysator og redusert ved 3,4 atm. H2 i 2 timer ved 23°C. Blandingen ble filtrert gjennom et lag "Dicalite" som deretter ble vasket med ytterligere 30 ml vann. Det fargeløse filtrat ble konsentrert under vakuum (40-45°C) til 50 ml. Løsningen ble fyllt på en 5 x 100 cm "CG-50" (NH4<+>) ionebytterkolonne. Etter vasking med 1000 ml vann ble ikke-reagert kanamycin A, 3-[l-(-)-y-amino-a-hydroksy-butyry lj-kanamycin A (BB-K29) og amikacin eluert med 0,5 N ammoniumhydroksyd. Polyacylmateriale ble utvunnet med 3N ammoniumhydroksyd. Bioanalyse, tynnsjiktskromatografi og optisk dreining ble anvendt for registrering av elueringens fremadskridelse. Volumet og den iakttatte optiske dreining for hver eluatfraksjon samt vekten og det prosentuelle utbytte av fast stoff isolert fra hver fraksjon ved inndamping til tørr tilstand er sammenfattet nedenfor: ml of water with a pH of 2.8. The combined aqueous fractions were washed with 50 mL of ethyl acetate. The solution was placed in a 500 ml Parr flask along with 5 g of 5% palladium-on-carbon catalyst and reduced at 3.4 atm. H2 for 2 hours at 23°C. The mixture was filtered through a layer of "Dicalite" which was then washed with another 30 ml of water. The colorless filtrate was concentrated under vacuum (40-45°C) to 50 ml. The solution was loaded onto a 5 x 100 cm "CG-50" (NH4<+>) ion exchange column. After washing with 1000 ml of water, unreacted kanamycin A, 3-[l-(-)-γ-amino-α-hydroxy-butyryl lj-kanamycin A (BB-K29) and amikacin were eluted with 0.5 N ammonium hydroxide. Polyacyl material was extracted with 3N ammonium hydroxide. Bioanalysis, thin-layer chromatography and optical rotation were used to record the progress of the elution. The volume and observed optical rotation for each eluate fraction as well as the weight and percentage yield of solid isolated from each fraction by evaporation to dryness are summarized below:
Det brukte dietylketonsjikt viste seg ved meget effektiv væskekromatografi å inneholde ytterligere 3-5% amikacin. The used diethyl ketone layer was shown by highly efficient liquid chromatography to contain an additional 3-5% amikacin.
Det rå amikacin (6,20 g) ble løst i 20 ml vann og tynnet med 20 ml metanol, og 20 ml isopropanol ble tilsatt for å frembringe krystallisasjon. Det ble oppnådd 6,0 g (45,8%) krystallinsk amikacin. The crude amikacin (6.20 g) was dissolved in 20 ml of water and diluted with 20 ml of methanol, and 20 ml of isopropanol was added to induce crystallization. 6.0 g (45.8%) of crystalline amikacin was obtained.
Eksempel 2 Example 2
Fremstilling av 1-N-[l-(-)-y-amino-a-hydroksybutyryl] -kana-* mycin A, amikacin, ved selektiv acylering av poly-(trimetylsilyl) 6'-N-karbobenzyloksykanamycin A i vannfri aceton. Preparation of 1-N-[1-(-)-γ-amino-α-hydroxybutyryl]-kanan-* mycin A, amikacin, by selective acylation of poly-(trimethylsilyl) 6'-N-carbobenzyloxykanamycin A in anhydrous acetone.
103 g (0,081 mol, regnet som 6<1->N-karbobenzyloksykanamycin 103 g (0.081 mol, calculated as 6<1->N-carbobenzyloxykanamycin
A (silyl) g) poly-(tr ime tyl silyl)-6 ' -N-karbobenzyloksy kanamycin A fremstilt som i eksempel 1 ble løst i 100 ml tørr aceton ved 23°C. 35,24 g (0,085 mol) L-(-)-y-benzyloksykarbonylamino-a-hydroksysmørsyre-N-hydroksy-5-norbornen-2,3-dikarboksimidester (NAE) løst i 100 ml tørr aceton ved 23°C ble langsomt tilsatt under god omrøring til løsningen av poly-(trimetylsilyl)-61 - N-karbobenzyloksykanamycin A i løpet av et tidsrom på 15 minutter. Løsningen ble omrørt ved 23°C i 20 timer under nitrogenatmosfære. Den blekgule, klare løsning (pH 7,2) ble tynnet med 100 ml vann. Blandingens pH-verdi ble innstilt på 2,5 (3N HCl), og omrøringen ble fortsatt ved 23°C i 15 minutter. Aceton ble fjernet under anvendelse av dampejektorvakuum ved ca. 3 5°C. Løsningen ble anbrakt i en 500 ml Parr-flaske sammen med 10 g 5% palladium-på-karbon katalysator (Engelhard) og redusert ved 2,72 atm. H2 i 2 timer ved 23 C. Blandingen ble filtrert gjennom et lag diatoméjord som deretter ble vasket med ytterligere 50 ml vann. Etter konsentrering til ca. 1/3 volum ble løsningen (pH 6,9-7,2) fyllt i en 6 x 110 cm "CG-50" (NH^<+>) ionebytterkolonne og eluert med en trinnvis gradient fra H20 til 0,6 N ammoniumhydroksyd for ut-vinnelse av amikacin. Et automatisk polarimeter ble anvendt for registrering av elueringens fremadskridelse. Kombinasjoner ble foretatt på basis av tynnsjiktskromatografivurdering. De kombinerte amikacinfraksjoner ble konsentrert til 25-30% tørrstoff. Løs-ningen ble tynnet med et like stort volum metanol, etterfulgt av 2 volumer isopropanol for å frembringe krystallisasjon. Det ble utvunnet 18,2 g (40%) krystallinsk amikacin. A (silyl) g) poly-(trimethylsilyl)-6'-N-carbobenzyloxy kanamycin A prepared as in example 1 was dissolved in 100 ml of dry acetone at 23°C. 35.24 g (0.085 mol) of L-(-)-γ-benzyloxycarbonylamino-α-hydroxybutyric acid-N-hydroxy-5-norbornene-2,3-dicarboximide ester (NAE) dissolved in 100 ml of dry acetone at 23°C was slowly added with good stirring to the solution of poly-(trimethylsilyl)-61-N-carbobenzyloxykanamycin A over a period of 15 minutes. The solution was stirred at 23°C for 20 hours under a nitrogen atmosphere. The pale yellow, clear solution (pH 7.2) was diluted with 100 ml of water. The pH of the mixture was adjusted to 2.5 (3N HCl), and stirring was continued at 23°C for 15 minutes. Acetone was removed using steam ejector vacuum at ca. 35°C. The solution was placed in a 500 ml Parr flask along with 10 g of 5% palladium-on-carbon catalyst (Engelhard) and reduced at 2.72 atm. H2 for 2 hours at 23 C. The mixture was filtered through a layer of diatomaceous earth which was then washed with another 50 ml of water. After concentration to approx. 1/3 volume solution (pH 6.9-7.2) was loaded into a 6 x 110 cm "CG-50" (NH^<+>) ion exchange column and eluted with a stepwise gradient from H 2 O to 0.6 N ammonium hydroxide for recovery of amikacin. An automatic polarimeter was used to record the progress of the elution. Combinations were made on the basis of thin-layer chromatography assessment. The combined amikacin fractions were concentrated to 25-30% solids. The solution was diluted with an equal volume of methanol, followed by 2 volumes of isopropanol to induce crystallization. 18.2 g (40%) of crystalline amikacin was recovered.
Utvinningen av 12% kanamycin A, 40% BB-K29 og 5% polyacylert kanamycin ga en materialbalanse på 97%. The recovery of 12% kanamycin A, 40% BB-K29 and 5% polyacylated kanamycin gave a material balance of 97%.
Eksempel 3 Example 3
Fremstilling av 1-N- [L-(-)-y-amino-a-hydroksybutyryl] - kanamycin A, amikacin, ved selektiv acylering av poly-(trimetylsilyl) -kanamycin A under anvendelse av blokkering in situ. Preparation of 1-N-[L-(-)-γ-amino-α-hydroxybutyryl]-kanamycin A, amikacin, by selective acylation of poly-(trimethylsilyl)-kanamycin A using in situ blocking.
A. Poly-( trimetylsilyl)- kanamycin A. A. Poly-(trimethylsilyl)-kanamycin A.
Kanamycin A fri base (18 g aktivitet, 37,15 mmol) ble oppslemmet i 200 ml tørr acetonitril og oppvarmet til tilbakekjøling. 29,8 g (184,6 mmol) heksametyldisilazan ble tilsatt i løpet av 30 minutter, og blandingen ble omrørt med tilbakekjøling i 78 timer, hvorved det-ble-oppnådd en__lysegul, klar løsning. Fjerning av løsningsmidlet under vakuum etterlot 43 g [94%, regnet som kanamycin A (silyl) 1Q] av en amorf-, fast rest. Kanamycin A free base (18 g activity, 37.15 mmol) was slurried in 200 ml dry acetonitrile and heated to reflux. 29.8 g (184.6 mmol) of hexamethyldisilazane was added over 30 minutes, and the mixture was stirred with reflux for 78 hours to give a pale yellow, clear solution. Removal of the solvent under vacuum left 43 g [94%, calculated as kanamycin A (silyl) 1Q ] of an amorphous solid residue.
B. 1-N- [.L- (-)- y- amino- g- hydroksybutyryl] - kanamycin A. B. 1-N- [.L- (-)- y- amino- g- hydroxybutyryl] - kanamycin A.
5,56 g (20,43 mmol) p-(benzyloksykarbonyloksy)-benzoesyre ble oppslemmet i 50 ml tørr acetonitril ved 23°C. 8,4 g (41,37 mmol) N,0-bis-trimetylsilylacetamid ble tilsatt under god omrøring. Løsningen ble holdt ved 23°C i 30 minutter og ble deretter i løpet av 3 timer under kraftig omrøring tilsatt til en løsning av 21,5 g (17,83 mmol, regnet som (silyl)1Q-forbindelsen) av poly-(trimetylsilyl)-kanamycin A i 7 5 ml tørr acetonitril ved 23°C. Blandingen ble omrørt i 4 timer, løsningsmidlet ble fjernet under vakuum (40°C), og den oljeaktige rest ble løst i 50 ml tørr aceton ved 23°C. 5.56 g (20.43 mmol) of p-(benzyloxycarbonyloxy)-benzoic acid was slurried in 50 ml of dry acetonitrile at 23°C. 8.4 g (41.37 mmol) of N,0-bis-trimethylsilylacetamide was added with good stirring. The solution was kept at 23°C for 30 minutes and was then added over the course of 3 hours with vigorous stirring to a solution of 21.5 g (17.83 mmol, calculated as the (silyl)1Q compound) of poly-(trimethylsilyl )-kanamycin A in 75 ml of dry acetonitrile at 23°C. The mixture was stirred for 4 hours, the solvent was removed under vacuum (40°C), and the oily residue was dissolved in 50 mL of dry acetone at 23°C.
8,55 g (20,63 mmol) L-(-)-y-benzyloksykarbonylamino-a-hydroksysmørsyre-N-hydroksy-5-norbornen-2,3-dikarbokimidester (NAE) i 30 ml aceton ble tilsatt til løsningen ovenfor i løpet av et tidsrom på 5 minutter. Blandingen ble holdt ved 23°C i 78 timer. Løsningen ble tynnet med 100 ml vann, og pH-verdien 8.55 g (20.63 mmol) of L-(-)-γ-benzyloxycarbonylamino-α-hydroxybutyric acid-N-hydroxy-5-norbornene-2,3-dicarboximide ester (NAE) in 30 ml of acetone was added to the above solution in during a period of 5 minutes. The mixture was kept at 23°C for 78 hours. The solution was diluted with 100 ml of water, and the pH value
(7,0) ble senket til 2,5 (6N HCl). Blandingen ble anbrakt i en 500 ml Parr-flaske sammen med 3 g 5% palladium-på-karbon katalysator (Engelhard) og redusert ved 2,7 2 atm. H2 i 2 timer ved 23°C. Blandingen ble filtrert gjennom et lag diatoméjord som deretter ble vasket med 20 ml vann. Det ble ved mikrobiologisk analyse overfor E. coli påvist at det kombinerte filtrat og vaskevann (168 ml) inneholdt ca. 11.400 mcg/ml (19% utbytte) amikacin. (7.0) was lowered to 2.5 (6N HCl). The mixture was placed in a 500 ml Parr flask along with 3 g of 5% palladium-on-carbon catalyst (Engelhard) and reduced at 2.7 2 atm. H2 for 2 hours at 23°C. The mixture was filtered through a layer of diatomaceous earth which was then washed with 20 ml of water. Microbiological analysis against E. coli showed that the combined filtrate and washing water (168 ml) contained approx. 11,400 mcg/ml (19% yield) amikacin.
Eksempel 4 Example 4
Fremstilling av 1-N-[L-(-)-y-amino-a-hydroksybutyryl] -kanamycin A, amikacin, ved selektiv acylering av poly-(trimetylsilyl)-kanamycin A. Preparation of 1-N-[L-(-)-γ-amino-α-hydroxybutyryl]-kanamycin A, amikacin, by selective acylation of poly-(trimethylsilyl)-kanamycin A.
A. Poly- ( trimetylsilyl)- kanamycin A. A. Poly-(trimethylsilyl)-kanamycin A.
En suspensjon av 10 g (20,6 mmol) kanamycin A i 100 ml tørr acetonitril og 25 ml (119 mmol) 1,1,1,3,3,3-heksametyldisilazan ble kokt med tilbakekjøling i 72 timer. Løsningen ble inndampet til tørr tilstand under vakuum ved 3 0-4 0°C. Det ble oppnådd 21,3 A suspension of 10 g (20.6 mmol) of kanamycin A in 100 ml of dry acetonitrile and 25 ml (119 mmol) of 1,1,1,3,3,3-hexamethyldisilazane was refluxed for 72 hours. The solution was evaporated to dryness under vacuum at 30-40°C. 21.3 was achieved
g poly-(trimetylsilyl)-kanamycin A som et lyst, gulbrunt, amorft pulver [85% utbytte regnet som kanamycin A (silyl)^g]• g poly-(trimethylsilyl)-kanamycin A as a light, yellowish-brown, amorphous powder [85% yield calculated as kanamycin A (silyl)^g]•
B. 1- N- LL-(-)- y- amino- a- hydroksybutyryl] - kanamycin A. B. 1- N- LL-(-)- y- amino- a- hydroxybutyryl] - kanamycin A.
Til en løsning av 2,4 g (2,0 mmol) poly-(trimetylsilyl)-kanamycin A i 3 0 ml tørr aceton ble det langsomt tilsatt 2,0 mmol L-(-)-y-benzyloksykarbonylamino-a-hydroksysmørsyre-N-hydroksy-5-norbornen-2,3-dikarboksimidester (NAE) i 10 ml tørr aceton ved 0-5°C. Reaksjonsblandingen ble omrørt ved 23°C i 1 uke og deretter inndampet til tø£r tilstand under vakuum ved en badtemperatur på 30-40°C. Det ble deretter tilsatt 60 ml vann til resten etterfulgt av 7 0 ml metanol for å frembringe en løsning. Løsningen ble surgjort med 3N HCl til pH 2,0 og deretter redusert ved 3,4 0 atm. H2 i 2 timer under anvendelse av 500 mg 5% palladium-på-karbon katalysator. Materialet ble filtrert, og det ble ved mikrobiologisk analyse overfor E. coli påvist at kombinert filtrat og vaskevæske inneholdt amikacin i et utbytte på 29,4%. To a solution of 2.4 g (2.0 mmol) of poly-(trimethylsilyl)-kanamycin A in 30 ml of dry acetone, 2.0 mmol of L-(-)-γ-benzyloxycarbonylamino-α-hydroxybutyric acid was slowly added N-hydroxy-5-norbornene-2,3-dicarboximide ester (NAE) in 10 ml of dry acetone at 0-5°C. The reaction mixture was stirred at 23°C for 1 week and then evaporated to dryness under vacuum at a bath temperature of 30-40°C. 60 ml of water was then added to the residue followed by 70 ml of methanol to produce a solution. The solution was acidified with 3N HCl to pH 2.0 and then reduced at 3.40 atm. H2 for 2 h using 500 mg of 5% palladium-on-carbon catalyst. The material was filtered, and it was demonstrated by microbiological analysis against E. coli that the combined filtrate and washing liquid contained amikacin in a yield of 29.4%.
Eksempel 5 Example 5
Fremstilling av amikacin ved selektiv N-acylering av polytrimetylsilyl- 6'- N- karbobenzoksykanamycin A i vannfri aceton. Preparation of amikacin by selective N-acylation of polytrimethylsilyl-6'-N-carbobenzoxykanamycin A in anhydrous acetone.
I. Sammendrag. I. Summary.
Silylering av 6'-N-karbobenzoksykanamycin A i acetonitril under anvendelse av heksametyldisilazan (HMDA) gir 6'-N-karbobenzoksykanamycin A (silyl)g-mellomproduktet © . Dette silylerte kanamycin A er lettløselig i de fleste organiske løsningsmidler. Acylering med NAE i vannfri aceton ved 23°C av et 5% molart overskudd av-NAE-i., f or ho ld til det anvendte 6'-N-Cbz kanamycin A ga en blanding som bare inneholdt Cbz-derivater av amikacin og BB-K29, noe uomsatt kanamycin A og noe polyacylmateriale. Det kunne ikke påvises BB-Kll ved noen av disse undersøkelser. Eluering av en acetonacyleringsblanding etter reduksjon og opp-arbeidelse fra en "CG-50" (NH4<+>) kolonne under anvendelse av en ammoniumhydroksydgradient ga isolerte utbytter av ren amikacin i området omkring 4 0%. Silylation of 6'-N-carbobenzoxykanamycin A in acetonitrile using hexamethyldisilazane (HMDA) gives the 6'-N-carbobenzoxykanamycin A (silyl)g intermediate © . This silylated kanamycin A is readily soluble in most organic solvents. Acylation with NAE in anhydrous acetone at 23°C of a 5% molar excess of -NAE-i., relative to the used 6'-N-Cbz kanamycin A gave a mixture containing only Cbz derivatives of amikacin and BB-K29, some unreacted kanamycin A and some polyacyl material. BB-Kll could not be detected in any of these examinations. Elution of an acetone acylation mixture after reduction and work-up from a "CG-50" (NH4<+>) column using an ammonium hydroxide gradient gave isolated yields of pure amikacin in the range of about 40%.
II. Reaksjonsskjemaer. A. III. Materialer II. Reaction forms. A. III. Materials
IV. Sikkerhet IV. Safety
6'- N- Cbz Kana A - Ingen direkte opplysninger tilgjengelig. Unngå støvkontakt. 6'- N- Cbz Kana A - No direct information available. Avoid contact with dust.
Acetonitril - Behandles som et cyanid. Unngå Acetonitrile - Treated as a cyanide. Avoid
innånding av damper. Kan forårsake hudirritasjon. inhalation of vapours. May cause skin irritation.
Heksametyldisilazan - Irritasjonsfremkallende, Hexamethyldisilazane - Irritant,
( HMDS) omgås med forsiktighet. (HMDS) should be handled with caution.
6'-N-Cbz Kana A (silyl)Ingen direkte opplysninger tilgjengelig, omgås med forsiktighet. 6'-N-Cbz Kana A (silyl) No direct information available, handle with caution.
BHBA - Toksisitet ikke fastslått. BHBA - Toxicity not determined.
Unngå eksponering for fast stoff. —HONB = Toksisitet ukjent. Utvis forsiktighet under håndtering. Avoid exposure to solids. —HONB = Toxicity unknown. Exercise caution during handling.
DCC - Alvorlig hud- og øye-irriterende. Unngå inhalering av tåke eller DCC - Severe skin and eye irritant. Avoid inhalation of mist or
damper. Toksisk. vapors. Toxic.
Aceton - Brennbar. Inhalering kan frem-kalle hodepine, tretthet, opp-hisselse, bronkial irritasjon og i store mengder narkose. Acetone - Flammable. Inhalation can cause headache, fatigue, agitation, bronchial irritation and, in large quantities, narcosis.
NAE - Ingen direkte informasjon tilgjengelig, anvendes alltid direkte NAE - No direct information available, always applied directly
som oppløsning i aceton. as a solution in acetone.
Metanol - Brennbar. Forgiftning kan inntreffe ved svelging, inhalering eller perkutan absorpsjon. Methanol - Flammable. Poisoning can occur by ingestion, inhalation or percutaneous absorption.
Isopropanol - Brennbar. Svelging eller inhalering av store mengder av dampene kan forårsake hodepine, svimmelhet, mental depresjon, oppkast, narkose. Isopropanol - Flammable. Swallowing or inhaling large quantities of the vapors may cause headache, dizziness, mental depression, vomiting, narcosis.
Ammoniumhydroksyd - Toksiske damper. Bær maske, unngå Ammonium hydroxide - Toxic fumes. Wear a mask, avoid
kontakt med væske. contact with liquid.
" CG- 50" ( NH. + ) - Ingen toksisitetsdata tilgjengelig, "CG- 50" ( NH. + ) - No toxicity data available,
omgås med forsiktighet. handled with care.
V. Fremgangsmåte V. Procedure
A. Fremstilling av 6'-N-karbobenzyloksykanamycin A (silyl)g A. Preparation of 6'-N-carbobenzyloxykanamycin A (silyl)g
[6'-N-Cbz Kana A (silyl) g] [6'-N-Cbz Kana A (silyl)g]
1. 50 g 6<1->N-karbobenzyloksykanamycin A (KF<4%) oppslemmes i 300 ml acetonitril (KF< 0,01%). Oppslemmingen bringes til koking ved tilbakekjøling (74°c) mens det opprettholdes en strøm av tørr nitrogen gjennom oppslemmingen. 2. I løpet av et tidsrom på 3 0 minutter tilsettes det langsomt 75,8 ml heksametyldisilazan (HMDS). Fullstendig opp-løsning vil inntreffe med utvikling av ammoniakkgass. 3. Tilbakekjøling fortsettes i 18-20 timer under nitrogengjennomblåsing. 4. Den klare, lysegule løsning konsentreres under vakuum (badtemperatur 4 0-50°C) til et skumliknende fast stoff. Utbytter av silylg-forbindelsen 89-92 g (90-94% av teoretisk utbytte). 1. 50 g of 6<1->N-carbobenzyloxykanamycin A (KF<4%) is suspended in 300 ml of acetonitrile (KF< 0.01%). The slurry is brought to boiling by reflux (74°c) while maintaining a flow of dry nitrogen through the slurry. 2. Over a period of 30 minutes, slowly add 75.8 ml of hexamethyldisilazane (HMDS). Complete dissolution will occur with the evolution of ammonia gas. 3. Recooling is continued for 18-20 hours under nitrogen purging. 4. The clear, pale yellow solution is concentrated under vacuum (bath temperature 40-50°C) to a foam-like solid. Yield of the silyl compound 89-92 g (90-94% of theoretical yield).
NB: For fremtidig referanse. I andre løsningsmiddelsunder-søkelser isoleres dette faste stoff vanligvis ikke, men anvendes direkte til acyleringen. NB: For future reference. In other solvent studies, this solid is usually not isolated, but used directly for the acylation.
B. Fremstilling av N-hydroksy-5-norbornen-2,3-dikarboksimidester av L-(-)-y-karbobenzyloksyamino-a-hydroksy-smørsyre ( NAE) B. Preparation of N-hydroxy-5-norbornene-2,3-dicarboximide ester of L-(-)-γ-carbobenzyloxyamino-α-hydroxybutyric acid (NAE)
1. 21,5 g L-(-)-y-karbobenzyloksyamino-a-hydroksysmørsyre (BHBA) løses i 100 ml tørr aceton ved 23°C og deretter 15,2 g N-hydroksy-5-norbornen-2,3-dikarboksimid (HOMB). Det oppnås en fullstendig oppløsning. 2. I løpet av 30 minutter tilsettes det en løsning av 17,48 g dicykloheksylkarbodiimid (DCC) i 50 ml aceton under omrøring. Temperaturen vil stige til ca. 4 0°C under tilsetningen ved utfelling av dicykloheksylurea (DCU). 3. Oppslemmingen omrører i 3-4 timer idet temperaturen tillates å utlignes til 23-25°C. 4. Ureaderivatet fjernes ved filtrering. Filterkaken vaskes med 30 ml aceton. Filtratet pluss vaskevæskene oppbevares til acyleringstrinnet nedenfor. 1. 21.5 g of L-(-)-γ-carbobenzyloxyamino-α-hydroxybutyric acid (BHBA) are dissolved in 100 ml of dry acetone at 23°C and then 15.2 g of N-hydroxy-5-norbornene-2,3- dicarboximide (HOMB). A complete resolution is achieved. 2. During 30 minutes, a solution of 17.48 g of dicyclohexylcarbodiimide (DCC) in 50 ml of acetone is added while stirring. The temperature will rise to approx. 4 0°C during the addition by precipitation of dicyclohexylurea (DCU). 3. The slurry is stirred for 3-4 hours while the temperature is allowed to equalize to 23-25°C. 4. The urea derivative is removed by filtration. The filter cake is washed with 30 ml of acetone. The filtrate plus the washings are kept for the acylation step below.
C. Acylering av 6'-N-Cbz Kana A (silyl)g. C. Acylation of 6'-N-Cbz Kana A (silyl)g.
1. 6'-N-Cbz Kana-A (silyl)g isolert under del A, trinn 4, løses i 100 ml tørr aceton ved 23-24 c. 2. Langsomt og under god omrøring tilsettes den under del B fremstilte NAE-løsning i løpet av et tidsrom på 15 minutter. Temperaturen vil gradvis stige til ca. 40°C. Løsningen tillates å komme i likevekt ved 23°C og omrøringen fortsettes i 18-20 timer under nitrogenatmosfære. 3. 100 ml vann tilsettes og pH-verdien (6,9-7,2) senkes til 2,2-2,5 med 6N saltsyre. Det omrøres i 15 minutter ved 23°C. (NB: Et ytterligere sjikt kan dannes. Dette gir ikke noe problem ved opparbeidelsen). 4. Aceton fjernes under vakuum ved en badtemperatur på 30-35°C. Konsentratet overføres til en passende hydrogeneringsbeholder (på forhånd gjennomblåst med nitrogen). 10 g 5% palladium-på-karbon katalysator tilsettes, og det hydrogeneres ved 2,72 atm. i 2-3 timer. 5. Blandingen filtreres gjennom et "Dicalite"-lag og hydrogeneringsbeholderen og filterkaken vaskes med ytterligere 5 0 ml vann. 6. Filtratet pluss vaskevæsken konsentreres til ca. 1/3 1. 6'-N-Cbz Kana-A (silyl)g isolated during part A, step 4, is dissolved in 100 ml of dry acetone at 23-24 c. 2. Slowly and with good stirring, add the NAE prepared during part B- solution within a period of 15 minutes. The temperature will gradually rise to approx. 40°C. The solution is allowed to equilibrate at 23°C and stirring is continued for 18-20 hours under a nitrogen atmosphere. 3. 100 ml of water is added and the pH value (6.9-7.2) is lowered to 2.2-2.5 with 6N hydrochloric acid. It is stirred for 15 minutes at 23°C. (NB: A further layer can be formed. This does not cause any problem during processing). 4. Acetone is removed under vacuum at a bath temperature of 30-35°C. The concentrate is transferred to a suitable hydrogenation vessel (pre-purged with nitrogen). 10 g of 5% palladium-on-carbon catalyst is added, and it is hydrogenated at 2.72 atm. for 2-3 hours. 5. The mixture is filtered through a "Dicalite" pad and the hydrogenation vessel and filter cake are washed with an additional 50 ml of water. 6. The filtrate plus the washing liquid is concentrated to approx. 1/3
av volumet (50 ml) under vakuum ved 4 0-4 5°C. of the volume (50 ml) under vacuum at 40-45°C.
7. pH-verdien kontrolleres. Den bør ligge i intervallet 6,9-7,2. Dersom ikke innstilles den med IN ammoniumhydroksyd. Blandingen fylles på en "CG-50" (NH4<+>) kolonne (6 x 110 cm). 8. Søylen vaskes med 1000 ml avionisert vann. Deretter elueres det med 0,5-0,6 N ammoniumhydroksyd under anvendelse av et automatisk polarimeter for registrering av elueringens fremad- ~ skridelse~. "Elueringsrekkefølgen er som følger: 7. The pH value is checked. It should be in the range 6.9-7.2. If not set it with IN ammonium hydroxide. The mixture is loaded onto a "CG-50" (NH4<+>) column (6 x 110 cm). 8. The column is washed with 1000 ml of deionized water. It is then eluted with 0.5-0.6 N ammonium hydroxide using an automatic polarimeter to record the progress of the elution. ~ progress~. "The elution order is as follows:
Det ble ikke påvist noe BB-Kll ved noen av acylerings-opparbeidelsene. Polyacylmateriale, dvs. 1,3-diAHBA-analogen til kana A, utvinnes ved utvasking av kolonnen med 3N ammoniumhydroksyd. 9. Amikacin-fraksjonene kombineres og konsentreres til 25-3 0% tørrstoff. Det tynnes med 1 volum metanol og podes med amikacinkrystaller. 10. Langsomt tilsettes det i løpet av 2 timer 2 volumer isopropanol (IPA) under god omrøring, og krystalliseres ved 23°C i 6-8 timer. 11. Det faste stoff frafiltreres, vaskes med 50 ml 1:1:2 vann/metanol/IPA-blanding og til slutt med 25 ml IPA. 12. Det tørkes under vakuum ved 40°C i 12-16 timer. Utbytte: 17,3-19,0 g (38-42%) amikacin med følgende egenskaper: No BB-Kll was detected in any of the acylation preparations. Polyacyl material, ie the 1,3-diAHBA analogue of kana A, is recovered by washing the column with 3N ammonium hydroxide. 9. The amikacin fractions are combined and concentrated to 25-30% solids. It is diluted with 1 volume of methanol and inoculated with amikacin crystals. 10. Slowly add 2 volumes of isopropanol (IPA) over 2 hours with good stirring, and crystallize at 23°C for 6-8 hours. 11. The solid is filtered off, washed with 50 ml of 1:1:2 water/methanol/IPA mixture and finally with 25 ml of IPA. 12. It is dried under vacuum at 40°C for 12-16 hours. Yield: 17.3-19.0 g (38-42%) amikacin with the following properties:
Tynnsj iktskromatografi. Thin layer chromatography.
CHCl3-metanol - NH^OH - vann (1:4:2:1), 5 x 20 cm silikagelplater fra Quantum Industries - én sone som påvist med nin-hydrin (RF~ 0,4). CHCl3-methanol - NH^OH - water (1:4:2:1), 5 x 20 cm silica gel plates from Quantum Industries - one zone as detected with nin-hydrin (RF~ 0.4).
Spesifikk dreining. Specific turning.
13. Utvinningen av BB-K29 i dette system var også 3 9-4 2%, restkana A 10-14% og 1,3-diAHBA-kana A ca. 5%, noe som gir en materialbalanse på over 95%. 13. The recovery of BB-K29 in this system was also 3 9-4 2%, residual kana A 10-14% and 1,3-diAHBA-kana A approx. 5%, which gives a material balance of over 95%.
Eksempel 6 Example 6
Fremstilling av amikacin ved selektiv N-acylering av poly-trlmetylsilyl kana A i vannfri aceton. Preparation of amikacin by selective N-acylation of polytrimethylsilyl kana A in anhydrous acetone.
I. Sammendrag. I. Summary.
Silylering av kana A "base" i acetonitril under anvendelse av heksametyldisilazan (HMDA) ga polytrimetylsilyl kana A. Silyleringsgraden er ennå uviss, men antas for tiden å være kana A (silyl)^q. Polysilylert kana A er lettløselig i de fleste organiske løsningsmidler. Acylering med SAE i vannfri aceton ved 23°C under anvendelse av et 1:1 molart forhold av SAE til anvendt kana A ga en blanding som inneholdt Cbz-derivater av amikacin og BB-K29, vanligvis i forholdet 2-3/1, BB-K6 (5-8%), uomsatt kana A (15-20%) samt noe polyacylmateriale (5-10%). Liksom det var tilfellet ved det tidligere arbeid med acylering av polytrimetylsilyl-6<1->N-karbobenzoksy kana A ble det ikke påvist noe BB-Kll ved noen av disse forsøk. Reduksjon og opp-arbeidelse av en acetonacyleringsblanding etterfulgt av kromato-grafi i en "CG-50"-(NH^<+>) kolonne under anvendelse av 0,5 N ammoniumhydroksyd ga isolert krystallinsk amikacin i området 34-39%. Silylation of kana A "base" in acetonitrile using hexamethyldisilazane (HMDA) gave polytrimethylsilyl kana A. The degree of silylation is still uncertain, but is currently believed to be kana A (silyl)^q. Polysilylated kana A is readily soluble in most organic solvents. Acylation with SAE in anhydrous acetone at 23°C using a 1:1 molar ratio of SAE to cana A used gave a mixture containing Cbz derivatives of amikacin and BB-K29, usually in a ratio of 2-3/1, BB -K6 (5-8%), unreacted kana A (15-20%) and some polyacyl material (5-10%). As was the case with the previous work on the acylation of polytrimethylsilyl-6<1->N-carbobenzoxycan A, no BB-Kll was detected in any of these experiments. Reduction and work-up of an acetone acylation mixture followed by chromatography on a "CG-50"-(NH₂<+>) column using 0.5 N ammonium hydroxide afforded isolated crystalline amikacin in the range of 34-39%.
II. Reaksjonsskjemaer. A. III. Materialer II. Reaction forms. A. III. Materials
IV. Sikkerhet IV. Safety
Kana A " base" - Kjent legemiddel - vanlig Kana A "base" - Known drug - common
forsiktighet tilrås. caution is advised.
Kana A (silyl) .. - Ingen direkte opplysninger tilgjengelig, omgås med forsiktighet. Kana A (silyl) .. - No direct information available, approach with caution.
Andre materialer - Se eksempel 5. Other materials - See example 5.
V. Fremgang små te V. Progress small teas
A. Fremstilling av Kana A (silyl)^q. A. Preparation of Kana A (silyl)^q.
1. 50 g kana A "base" (KF 2,5-3,5%) oppslemmes i 500 ml aceton (KF<0,01%). Oppslemmingen bringes til tilbakekjøling (74°C) mens det opprettholdes en strøm av tørr nitrogen gjennom oppslemmingen. 2. I løpet av 3 0 minutter tilsettes det langsomt 112 ml heksametyldisilazan (HMDS). Fullstendig oppløsning vil inntreffe innen 4-5 timer med utvikling av ammoniakkgass. 3. Koking med tilbakekjøling fortsettes i 22-26 timer under nitrogengjennomblåsing. 4. Den klare, svakt gule løsning konsentreres under vakuum (40°C) til en tyktflytende rest. Det skylles med ytterligere 100 ml acetonitril og tørkes fullstendig under høyvakuum i 3-6 timer. Utbytte av hvitaktig, amorft, fast stoff er 109-115 g (90-95% av det teoretiske, regnet som kana A (silyl)^q)• B. Fremstilling av N-hydroksysuccinimidester av L-(-)-a-karbobenzyloksyamino- ct- hydroksysmørsyre ( SAE. 1. 100 g L-(-)-a-benzyloksykarbonylamino-a-hydroksysmørsyre (BHBA) og 4 5,38 g N-hydroksysuccinimid (N-HOS) løses i 13 00 ml etylacetat (KF<0,05%) under omrøring ved 23°C. 2. 81,29 g dicykloheksylkarbodiimid (DCC) løses i 400 ml 1. 50 g kana A "base" (KF 2.5-3.5%) is suspended in 500 ml acetone (KF<0.01%). The slurry is brought to reflux (74°C) while maintaining a flow of dry nitrogen through the slurry. 2. Over 30 minutes, slowly add 112 ml of hexamethyldisilazane (HMDS). Complete dissolution will occur within 4-5 hours with evolution of ammonia gas. 3. Boiling with recooling is continued for 22-26 hours under nitrogen purging. 4. The clear, slightly yellow solution is concentrated under vacuum (40°C) to a viscous residue. It is rinsed with a further 100 ml of acetonitrile and dried completely under high vacuum for 3-6 hours. Yield of whitish, amorphous solid is 109-115 g (90-95% of the theoretical, calculated as kana A (silyl)^q)• B. Preparation of N-hydroxysuccinimide esters of L-(-)-a-carbobenzyloxyamino - ct- hydroxybutyric acid ( SAE. 1. 100 g of L-(-)-a-benzyloxycarbonylamino-a-hydroxybutyric acid (BHBA) and 4 5.38 g of N-hydroxysuccinimide (N-HOS) are dissolved in 13 00 ml of ethyl acetate (KF< 0.05%) with stirring at 23° C. 2. Dissolve 81.29 g of dicyclohexylcarbodiimide (DCC) in 400 ml
etylacetat (KF<0,05%) ved 23°C. Under god omrøring tilsettes denne løsning i løpet av 3 0 minutter til løsningen fra trinn 1. Temperaturen vil stige til 4 0-42°C under samtidig utfelling av dicykloheksylurea (DCU). ethyl acetate (KF<0.05%) at 23°C. With good stirring, this solution is added over 30 minutes to the solution from step 1. The temperature will rise to 40-42°C during simultaneous precipitation of dicyclohexylurea (DCU).
Oppslemmingen omrøres i 3-4 timer hvorved temperaturen tillates å likevektsinnstilles på 23°C. 3. DCU f raf Utreres og filterkaken vaskes med 250 ml etylacetat (KF< 0,05%). DCU-kaken kastes mens filtratet og vaskevæskene oppbevares. The slurry is stirred for 3-4 hours whereby the temperature is allowed to equilibrate to 23°C. 3. DCU f raf Drain and wash the filter cake with 250 ml ethyl acetate (KF< 0.05%). The DCU cake is discarded while the filtrate and washing liquids are kept.
4. Filtratet pluss vaskevæskene konsentreres til ca. 500 4. The filtrate plus the washing liquids are concentrated to approx. 500
ml (under vakuum ved 3 0-3 5°C). Litt produkt vil utkrystalliseres. ml (under vacuum at 30-35°C). A little product will crystallize out.
5. Konsentratet overføres til en passende beholder og under kraftig omrøring tilsettes det 100 ml heptan. Om nødvendig tilsettes SAE-podekrystaller. Krystallisasjon vil begynne nesten øyeblikkelig. Oppslemmingen omrøres i 30 minutter ved 23°C. 6. I løpet av 30 minutter tilsettes det 400 ml heptan, og oppslemmingen omrøres 4-5 timer ved 23°C. 7. Kaken filtreres og vaskes med 200 ml 3:1 heptan/etylacetat etterfulgt av 100 ml heptan. 8. Det tørkes i en vakuum-ovn ved 30-35°C i 18-20 timer. 5. The concentrate is transferred to a suitable container and, with vigorous stirring, 100 ml of heptane is added. If necessary, SAE seed crystals are added. Crystallization will begin almost immediately. The slurry is stirred for 30 minutes at 23°C. 6. Over the course of 30 minutes, 400 ml of heptane are added, and the slurry is stirred for 4-5 hours at 23°C. 7. The cake is filtered and washed with 200 ml of 3:1 heptane/ethyl acetate followed by 100 ml of heptane. 8. It is dried in a vacuum oven at 30-35°C for 18-20 hours.
Utbyttet er 110,1-131,4 g (80-95%). The yield is 110.1-131.4 g (80-95%).
Smp. : 119-120°C med mykning ved 114°C (korr.) Tynnsjiktskromatografi: 4 aceton:12 benzen:l CH^CO-H - på-visning 1% vandig KMO^. Temp. : 119-120°C with softening at 114°C (corr.) Thin layer chromatography: 4 acetone:12 benzene:1 CH^CO-H - on display 1% aqueous KMO^.
Rf: 0,7 for SAE, 0,2 BHBA på 2 x 10 cm formerkete silikagelplater fra Analtech Inc. Rf: 0.7 for SAE, 0.2 BHBA on 2 x 10 cm pre-coated silica gel plates from Analtech Inc.
C. Acylering av Kana A (silyl) ^q. C. Acylation of Kana A (silyl) ^q.
1. Det under avsnitt A, trinn 4, isolerte kana A (silyl) løses i 100 ml tørr aceton ved 23°C. 1. The kana A (silyl) isolated under section A, step 4 is dissolved in 100 ml of dry acetone at 23°C.
2. Hurtig og under god omrøring tilsettes det under avsnitt 2. Quickly and with good stirring, it is added under section
B fremstilte SAE (35,03 g) som en 10 prosentig løsning i tørr aceton i løpet av 5-10 minutter. Temperaturen vil stige ca. 5°C. Løsningen tillates å likevektsinnstilles på 23°C, og omrøring fortsettes i 18-20 timer. B prepared SAE (35.03 g) as a 10 percent solution in dry acetone over 5-10 minutes. The temperature will rise approx. 5°C. The solution is allowed to equilibrate at 23°C, and stirring is continued for 18-20 hours.
3. Den lysorange, klare løsning tynnes med 400 ml vann, 3. The light orange, clear solution is diluted with 400 ml of water,
og pH-verdien (7,0-7,5) senkes til 2,2-2,5 med 3N saltsyre. and the pH value (7.0-7.5) is lowered to 2.2-2.5 with 3N hydrochloric acid.
Den klare løsning omrøres nå ved 23°C i 15-30 minutter. The clear solution is now stirred at 23°C for 15-30 minutes.
4. Aceton fjernes under vakuum ved en badtemperatur på 3 0-35°C (en liten materialmengde kan utskilles på dette tidspunkt, men betyr ikke noe problem). Konsentratet overføres til en passende hydrogeneringsbeholder. Det tilsettes 10 g palladium-på-karbon katalysator, og det hydrogeneres ved 3,4 0 atm. i 2-3 timer. 5. Blandingen filtreres gjennom et "Dicalite"-lag, og hydrogeneringsbeholderen og filterkaken vaskes med ytterligere 4. Acetone is removed under vacuum at a bath temperature of 30-35°C (a small amount of material may separate at this point, but does not pose a problem). The concentrate is transferred to a suitable hydrogenation vessel. 10 g of palladium-on-carbon catalyst are added and hydrogenated at 3.40 atm. for 2-3 hours. 5. The mixture is filtered through a "Dicalite" layer, and the hydrogenation vessel and filter cake are washed with additional
2 x 50 ml vann. 2 x 50 ml water.
6. Filtratet pluss vaskevæskene konsentreres til ca en tredjedel av volumet (150-165 ml) under vakuum ved 40-45°C. 7. pH-verdien ligger på dette tidspunkt i intervallet 6,0-7,0. Blandingen helles i en "CG-50" (NH4<+>) kolonne (6 x 110 cm). 8. Kolonnen vaskes med 1000 ml avionisert vann. Det elueres med 0,5 N ammoniumhydroksyd under anvendelse av et automatisk polarimeter for overvåkning av elueringens fremadskridelse. Elueringsrekkefølgen er som følger: 6. The filtrate plus the washing liquids are concentrated to about a third of the volume (150-165 ml) under vacuum at 40-45°C. 7. The pH value is at this point in the range 6.0-7.0. The mixture is poured into a "CG-50" (NH4<+>) column (6 x 110 cm). 8. The column is washed with 1000 ml of deionized water. It is eluted with 0.5 N ammonium hydroxide using an automatic polarimeter to monitor the progress of the elution. The elution order is as follows:
Rest kana A BB-K6 BB-K29 amikacin. Rest kana A BB-K6 BB-K29 amikacin.
Det ble ikke påvist noe BB-Kll ved noen av forsøkene. No BB-Kll was detected in any of the experiments.
9. Amikacinfraksjonene kombineres og konsentreres til 25-3 0% tørrstoff. Det tynnes med 1 volum metanol og podes med amikacinkrystaller. 10. I løpet av 2 timer tilsettes det langsomt 2 volumer IPA under god omrøring, og det krystalliseres ved 23°C i 6-8 timer. 11. Det faste stoff filtreres og vaskes med 35 ml 1:1:2 vann/metanol/IPA og til slutt med 3 5 ml IPA. 12. Det tørkes i en vakuum-ovn ved 40°C i 16-24 timer. 9. The amikacin fractions are combined and concentrated to 25-30% dry matter. It is diluted with 1 volume of methanol and inoculated with amikacin crystals. 10. Over the course of 2 hours, 2 volumes of IPA are slowly added with good stirring, and it crystallizes at 23°C for 6-8 hours. 11. The solid is filtered and washed with 35 ml of 1:1:2 water/methanol/IPA and finally with 35 ml of IPA. 12. It is dried in a vacuum oven at 40°C for 16-24 hours.
Utbytte: 19,91-22,84 g (34-39%). IR-, PMR- og CMR-spektral-data samt spesifikk dreining var fullstendig<1>overensstemmelse med den ønskete struktur. Yield: 19.91-22.84 g (34-39%). IR, PMR and CMR spectral data as well as specific rotation were in complete <1> agreement with the desired structure.
Tynnsjiktskromatografisystem Thin layer chromatography system
CHCl3/metanol/NH4OH/vann (1:4:2:1) 5 x 20 cm silikagelplater fra Quantum Industries, 1 sone amikacin med Rf~ 0,4 (ninhydrin-påvisning). CHCl3/methanol/NH4OH/water (1:4:2:1) 5 x 20 cm silica gel plates from Quantum Industries, 1 zone amikacin with Rf~ 0.4 (ninhydrin detection).
Eksempel 7 Example 7
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl) -6 ' -N-Cbz-kana A i tetrahydrofuran med blandet syreanhydrid av pivalinsyre og BHBA. Preparation of amikacin by acylation of poly-(trimethylsilyl)-6'-N-Cbz-cana A in tetrahydrofuran with mixed acid anhydride of pivalic acid and BHBA.
A. Fremstilling av blandet anhydrid. A. Preparation of mixed anhydride.
5,066 g (20,0 mmol) BHBA, 4,068 g (20,0 mmol) BSA og 2,116 g (22,0 mmol) etylamin ble løst i 200 ml sikttørket tetrahydrofuran. Løsningen ble kokt med tilbakekjøling i 2 1/4 time og deretter avkjølt til -10°C. 2,412 g (20,0 mmol) pivaloylklorid ble tilsatt i løpet av 2-3 minutter under omrøring, og omrøringen ble fortsatt i 2 timer ved -10°C. Temperaturen fikk deretter stige til 23°C. 5.066 g (20.0 mmol) of BHBA, 4.068 g (20.0 mmol) of BSA and 2.116 g (22.0 mmol) of ethylamine were dissolved in 200 ml of sieve-dried tetrahydrofuran. The solution was refluxed for 2 1/4 hours and then cooled to -10°C. 2.412 g (20.0 mmol) of pivaloyl chloride was added over 2-3 minutes with stirring, and stirring was continued for 2 hours at -10°C. The temperature was then allowed to rise to 23°C.
B. Acylering av poly-( trimetylsilyl)- 6'- N- Cbz kana A. B. Acylation of poly-(trimethylsilyl)-6'-N-Cbz kana A.
5,454 g (4,97 mmol, regnet som 6'-N-Cbz kana A (silyl)g) poly-(trimetylsilyl)-6<1->N-Cbz kana A fremstilt som i eksempel 1 ble løst i 50 ml tørr (molekylsikt) tetrahydrofuran ved 23°C. Halvparten av den i trinn A ovenfor fremstilte løsning av 5.454 g (4.97 mmol, calculated as 6'-N-Cbz kana A (silyl)g) poly-(trimethylsilyl)-6<1->N-Cbz kana A prepared as in example 1 was dissolved in 50 ml of dry (molecular sieve) tetrahydrofuran at 23°C. Half of the solution prepared in step A above
blandet anhydrid (10,0 mmol) ble tilsatt i løpet av 20 minutter under omrøring, og omrøringen ble fortsatt i 7 dager. mixed anhydride (10.0 mmol) was added over 20 min with stirring, and stirring was continued for 7 days.
Deretter ble det tilsatt 100 ml vann til reaksjonsblandingen, og pH (5,4) ble innstilt på 2,0 med 3 M H2S04- Omrøring ble fortsatt i 1 time, og løsningen ble ekstrahert med etylacetat. Polyacylert materiale begynte å krystallisere ut, så reaksjonsblandingen ble filtrert. Etter tørking over P2°5 veiet det ut~ vunnete faste stoff 0,702 g. Ekstraksjonen av reaksjonsblandingen ble fortsatt i totalt 4 x 7 5 ml etylacetat, hvoretter overskytende etylacetat ble drevet bort fra det vandige sjikt. En aliquot av den vandige løsning ble underkastet analyse ved hjelp av HPLC. Den resulterende kurve indikerte et utbytte på 26,4% av di-Cbz amikacin. Then 100 mL of water was added to the reaction mixture, and the pH (5.4) was adjusted to 2.0 with 3 M H 2 SO 4 - Stirring was continued for 1 hour, and the solution was extracted with ethyl acetate. Polyacylated material began to crystallize out, so the reaction mixture was filtered. After drying over P2°5, the recovered solid weighed 0.702 g. The extraction of the reaction mixture was continued in a total of 4 x 75 ml of ethyl acetate, after which excess ethyl acetate was driven away from the aqueous layer. An aliquot of the aqueous solution was subjected to analysis by HPLC. The resulting curve indicated a 26.4% yield of di-Cbz amikacin.
Det vandige sjikt ble deretter hydrogenert i et Parr-apparat ved 3,4 0 atm. H2~trykk i 2 timer under anvendelse av 0,5 g 10% Pd-på-karbon katalysator. Materialet ble filtrert, og det ble påvist overfor E. coli at kombinert filtrat og vaskevæsker inneholdt et 31,2% utbytte av amikacin. Amikacin/BB-K29-forholdet var ca. 9-10:1. Spor av polyacyl og ureagert kana A forelå. The aqueous layer was then hydrogenated in a Parr apparatus at 3.40 atm. H 2 ~ pressure for 2 hours using 0.5 g of 10% Pd-on-carbon catalyst. The material was filtered, and it was demonstrated against E. coli that the combined filtrate and washing liquids contained a 31.2% yield of amikacin. The amikacin/BB-K29 ratio was approx. 9-10:1. Traces of polyacyl and unreacted kana A were present.
Eksempel 8 Example 8
Innvirkning av vann for fremstillingen av amikacin ved acylering av poly-(trimetylsilyl)-kana A i acetonløsning ved 23°C. Effect of water on the production of amikacin by acylation of poly-(trimethylsilyl)cana A in acetone solution at 23°C.
A. Vannfritt løsningsmiddel. A. Anhydrous solvent.
2,40 g (2,0 mmol, regnet som kana A (silyl)1Q) poly-(trimetylsilyl) kana A fremstilt som i eksempel 3 ble løst i 20 ml aceton som var blitt tørket med en molekylsikt. Løsningen ble omrørt ved 23°C, og en løsning av 0,701 g (2,0 mmol) SAE i 10 ml sikttørket aceton ble tilsatt i løpet av 10 sekunder. Omrøring ble fortsatt ved 23°C i 22 timer. Det ble tilsatt 50 ml vann, 2.40 g (2.0 mmol, calculated as kana A (silyl)1Q) poly-(trimethylsilyl) kana A prepared as in example 3 was dissolved in 20 ml of acetone which had been dried with a molecular sieve. The solution was stirred at 23°C, and a solution of 0.701 g (2.0 mmol) SAE in 10 mL sieve-dried acetone was added over 10 seconds. Stirring was continued at 23°C for 22 hours. 50 ml of water was added,
og pH (7,5) ble innstilt på 2,5. Acetonen ble avdampet under vakuum ved 4 0°C, og den vandige løsning ble deretter redusert ved et H2~trykk på 3,47 atm. ved 23°C i 2 timer under "anvendelse av 1,0 g 10% Pd-på-karbon som katalysator. Mikrobiologisk analyse viste et 31,24% utbytte av amikacin. and pH (7.5) was adjusted to 2.5. The acetone was evaporated under vacuum at 40°C, and the aqueous solution was then reduced at a H 2 ~ pressure of 3.47 atm. at 23°C for 2 hours using 1.0 g of 10% Pd-on-carbon as catalyst. Microbiological analysis showed a 31.24% yield of amikacin.
B. Løsningsmiddel tilsatt vann. B. Solvent added to water.
Trinn A ovenfor ble gjentatt, bortsett fra at 1,0 ml (56 mmol) vann ble tilsatt til løsningen av poly-(trimetylsilyl)-kana A, Step A above was repeated, except that 1.0 mL (56 mmol) of water was added to the solution of poly-(trimethylsilyl)cana A,
og det ble omrørt i 15 minutter før acylering med SAE. Mikrobiologisk analyse viste et utbytte på 33,80% amikacin. and it was stirred for 15 min before acylation with SAE. Microbiological analysis showed a yield of 33.80% amikacin.
Eksempel 9 Example 9
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl) -6 1 -N-Cnz kana A i aceton med det blandete anhydrid av BHBA og isobutylkarbonsyre. Preparation of amikacin by acylation of poly-(trimethylsilyl)-6 1 -N-Cnz kana A in acetone with the mixed anhydride of BHBA and isobutylcarbonic acid.
A. Fremstilling av blandet anhydrid. A. Preparation of mixed anhydride.
1,267 g (5,0 mmol) BHBA og 1,313 g (10,0 mmol) N-trimetylsilylacetamid (MSA) i 20 ml sikttørket aceton ble omrørt ved 23°C, og 0,70 ml (5,0 mmol) trietylamin (TEA) ble tilsatt. Blandingen ble kokt med tilbakekjøling under ^-atmosfære i 2\ time. Blandingen ble avkjølt til -20°C, og 0,751 g (0,713 ml, 5,50 mmol) isobutylklorformiat ble tilsatt. Trietylaminhydroklorid begynte umiddelbart å utskilles. Blandingen ble omrørt i 1 time ved -20°C. 1.267 g (5.0 mmol) BHBA and 1.313 g (10.0 mmol) N-trimethylsilylacetamide (MSA) in 20 ml sieve-dried acetone were stirred at 23°C, and 0.70 ml (5.0 mmol) triethylamine (TEA ) was added. The mixture was refluxed under a 2-atmosphere for 2 hours. The mixture was cooled to -20°C and 0.751 g (0.713 mL, 5.50 mmol) of isobutyl chloroformate was added. Triethylamine hydrochloride immediately began to be excreted. The mixture was stirred for 1 hour at -20°C.
B. Acylering. B. Acylation.
6,215 g (4,9 mmol, regnet som (silyl)^-forbindelsen) poly-(trimetylsilyl)-6'-N-Cbz-kana A fremstilt som i eksempel 1 ble løst i 20 ml sikttørket aceton under omrøring ved 23°C. Løsningen ble avkjølt til -20°C, og den kalde løsning av blandet anhydrid fra trinn A ble langsomt tilsatt i løpet av 30 minutter. Reaksjonsblandingen ble omrørt i ytterligere lh time ved -20°C og deretter i 17 timer ved 23°C. Reaksjonsblandingen ble deretter helt i 150 ml vann ved 23°C under omrøring, pH (7,75) innstilt på 2,5 med 3N HCl, og omrøring ble fortsatt i 15 minutter. Aceton ble deretter avdampet under vakuum ved 4 0°C. En aliquot av den resulterende vandige løsning ble underkastet analyse ved hjelp av HPLC. Den resulterende kurve viste et utbytte på 34,33% di-Cbz-amikacin. 6.215 g (4.9 mmol, calculated as the (silyl)^-compound) poly-(trimethylsilyl)-6'-N-Cbz-cana A prepared as in example 1 was dissolved in 20 ml sieve-dried acetone with stirring at 23°C . The solution was cooled to -20°C and the cold solution of mixed anhydride from step A was slowly added over 30 minutes. The reaction mixture was stirred for a further 1h hour at -20°C and then for 17 hours at 23°C. The reaction mixture was then poured into 150 mL water at 23°C with stirring, pH (7.75) adjusted to 2.5 with 3N HCl, and stirring continued for 15 minutes. Acetone was then evaporated under vacuum at 40°C. An aliquot of the resulting aqueous solution was subjected to analysis by HPLC. The resulting curve showed a yield of 34.33% di-Cbz-amikacin.
Hovedparten av den vandige løsning ble redusert ved 3,4 0 atm. H2~trykk ved 23°C i 3 1/4 time under anvendelse av 2,0 g Pd/C katalysator. Katalysatoren ble fjernet ved filtrering, og ved mikrobiologisk analyse overfor E. coli ble det påvist at kombinert filtrat og vaskevæsker inneholdt et utbytte på 3 5,0% amikacin. The bulk of the aqueous solution was reduced at 3.40 atm. H 2 ~ pressure at 23°C for 3 1/4 hours using 2.0 g of Pd/C catalyst. The catalyst was removed by filtration, and by microbiological analysis against E. coli it was demonstrated that the combined filtrate and washing liquids contained a yield of 35.0% amikacin.
Eksempel 10 Example 10
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl)- 6'- N- Cbz- kana A i 3- pentanon. Preparation of amikacin by acylation of poly-(trimethylsilyl)-6'-N-Cbz-cana A in 3-pentanone.
30 g (23,65 mmol, regnet som 6'-N-Cbz-kana A (silyl)g) poly-(trimetylsilyl)-6'-N-Cbz-kana fremstilt som i eksempel 1 30 g (23.65 mmol, calculated as 6'-N-Cbz-cana A (silyl)g) poly-(trimethylsilyl)-6'-N-Cbz-cana prepared as in example 1
og løst i 100 ml sikttørket 3-pentanon ble omrørt ved 23°C, og NAE (26,02 mmol, 10% overskudd) ble tilsatt i løpet av 40 minutter. Omrøring ble fortsatt i 113 timer ved 23°C, og blandingen ble deretter tilsatt til 250 ml vann under kraftig omrøring. pH (7,3) ble innstilt på 2,5 med 3N HCl, hvoretter blandingen ble omrørt i ytterligere 3 0 minutter og 3-pentanon avdrevet under vakuum ved 40°C. Den vandige løsning ble ekstrahert med 4 x 100 ml etylacetat. En aliquot av den vandige løsning ble deretter underkastet analyse ved hjelp av HPLC. Den resulterende kurve viste et utbytte på 4 6,12% di-Cbz-amikacin. and dissolved in 100 mL of sieve-dried 3-pentanone was stirred at 23°C, and NAE (26.02 mmol, 10% excess) was added over 40 minutes. Stirring was continued for 113 hours at 23°C, and the mixture was then added to 250 ml of water with vigorous stirring. The pH (7.3) was adjusted to 2.5 with 3N HCl, after which the mixture was stirred for a further 30 minutes and 3-pentanone driven off under vacuum at 40°C. The aqueous solution was extracted with 4 x 100 ml of ethyl acetate. An aliquot of the aqueous solution was then subjected to analysis by HPLC. The resulting curve showed a yield of 4 6.12% di-Cbz-amikacin.
Hovedparten av den vandige reaksjonsblanding ble redusert ved 3,47 atm. H2_trykk ved 23°C i 2\ time under anvendelse av 3,0 g 10% Pd/C katalysator. Mikrobiologisk analyse av en aliquot av kombinert filtrat og vaskevæsker viste et utbytte på 4 0,24% amikacin. Hovedparten av den reduserte, vandige reaksjonsblanding ble deretter konsentrert under vakuum ved 40°C til ca. 100 ml og fraksjonert i en "CG-50" (NH4<+>) ionebytterkolonne (ca. 10 cm x ca. 120 cm, inneholdende ca. 10 liter harpiks). Den vandige løsning ble helt i kolonnen som ble vasket med 5 liter vann, og materialet ble eluert med 0,5N NH^OH, etterfulgt av 3N NH^OH The bulk of the aqueous reaction mixture was reduced at 3.47 atm. H 2 -pressure at 23°C for 2 hours using 3.0 g of 10% Pd/C catalyst. Microbiological analysis of an aliquot of combined filtrate and washings showed a yield of 4 0.24% amikacin. The bulk of the reduced aqueous reaction mixture was then concentrated under vacuum at 40°C to approx. 100 ml and fractionated in a "CG-50" (NH4<+>) ion exchange column (approx. 10 cm x approx. 120 cm, containing approx. 10 liters of resin). The aqueous solution was poured into the column which was washed with 5 liters of water, and the material was eluted with 0.5N NH^OH, followed by 3N NH^OH
for eluering av polyacylerte produkter. Polarimetri av fraksjonene viste et utbytte på 4 2,7% amikacin, et utbytte på 12,0% ureagert kanamycin A, et utbytte på 12,4% polyacylert materiale og et utbytte på 23,2% BB-K29. for elution of polyacylated products. Polarimetry of the fractions showed a yield of 4 2.7% amikacin, a 12.0% yield of unreacted kanamycin A, a 12.4% yield of polyacylated material and a 23.2% yield of BB-K29.
Eksempel 11 Example 11
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl) -6 ' -N-Cbz-kana A i vannfri cykloheksanon i varierende tidsrom. Preparation of amikacin by acylation of poly-(trimethylsilyl)-6'-N-Cbz-kana A in anhydrous cyclohexanone for varying periods of time.
A. 2,537 g (2,0 mmol, regnet som 6<1->N-Cbz-kana A (silyl)g) poly-(trimetylsilyl)-6'-N-Cbz-kana A fremstilt som i eksempel 1 A. 2.537 g (2.0 mmol, calculated as 6<1->N-Cbz-cana A (silyl)g) poly-(trimethylsilyl)-6'-N-Cbz-cana A prepared as in example 1
og løst i 3 00 ml tørr cykloheksanon ble acylert i 20 timer ved 23°C med en NAE-løsning i tørr cykloheksanon (10,8 ml av en 0,1944 mmol/ml løsning, 2,10 mmol). Reaksjonsblandingen ble deretter tilsatt til 150 ml vann under omrøring, og pH (5,6) ble innstilt på 2,5 med 3N HCl. Cykloheksanonen ble avdampet under and dissolved in 300 mL of dry cyclohexanone was acylated for 20 h at 23°C with a NAE solution in dry cyclohexanone (10.8 mL of a 0.1944 mmol/mL solution, 2.10 mmol). The reaction mixture was then added to 150 mL of water with stirring, and the pH (5.6) was adjusted to 2.5 with 3N HCl. The cyclohexanone was evaporated under
vakuum ved 4 0°C, og en aliquot av den resterende vandige fase ble tatt ut for analyse ved hjelp av HPLC. vacuum at 40°C, and an aliquot of the remaining aqueous phase was removed for analysis by HPLC.
Hovedparten av den vandige fase ble redusert under 3,4 0 atm. H2-trykk i 3 timer ved 23°C under anvendelse av 1,0 g 10% Pd/C katalysator. Katalysatoren ble fjernet ved filtrering, og kombinert filtrat og vaskevæsker ble analysert mikrobiologisk på amikacin. The majority of the aqueous phase was reduced below 3.40 atm. H 2 pressure for 3 hours at 23°C using 1.0 g of 10% Pd/C catalyst. The catalyst was removed by filtration, and the combined filtrate and washings were analyzed microbiologically for amikacin.
B. Reaksjon A ble gjentatt, bortsett fra at acyleringen ble fortsatt i 115 timer istedenfor 20 timer. B. Reaction A was repeated, except that the acylation was continued for 115 hours instead of 20 hours.
Eksempel 12 Example 12
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl) -6 ' -N-Cbz-kana A i vannfri tetrahydrofuran i varierende tidsrom. Preparation of amikacin by acylation of poly-(trimethylsilyl)-6'-N-Cbz-kana A in anhydrous tetrahydrofuran for varying periods of time.
A. Eksempel 11 A ble gjentatt, bortsett fra at det som løs-ningsmiddel istedenfor tørr cykloheksanon ble anvendt tørr tetra-hydrof uran. A. Example 11 A was repeated, except that dry tetrahydrofuran was used as solvent instead of dry cyclohexanone.
B. Eksempel 11 B ble gjentatt, bortsett fra at det som løs-ningsmiddel istedenfor tørr cykloheksanon ble anvendt tørr tetra-hydrof uran. B. Example 11 B was repeated, except that dry tetrahydrofuran was used as solvent instead of dry cyclohexanone.
Eksempel 13 Example 13
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl) -6 1 -N-Cbz-kana A i vannfri dioksan i varierende tidsrom. Preparation of amikacin by acylation of poly-(trimethylsilyl)-6 1 -N-Cbz-kana A in anhydrous dioxane for varying periods of time.
A. Eksempel 11 A ble gjentatt, bortsett fra at acyleringen ble fortsatt i 44 timer under anvendelse av tørr dioksan som løsningsmiddel. A. Example 11 A was repeated, except that the acylation was continued for 44 hours using dry dioxane as the solvent.
B. Eksempel 11 B ble gjentatt, bortsett fra at acyleringen ble fortsatt i 18^ time under anvendelse av tørr dioksan som løsningsmiddel. B. Example 11 B was repeated, except that the acylation was continued for 18 hours using dry dioxane as the solvent.
Eksempel 14 Example 14
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl)- 6'- N- Cbz- kana A i vannfri dietylketon ved 75°C. Preparation of amikacin by acylation of poly-(trimethylsilyl)-6'-N-Cbz-cana A in anhydrous diethyl ketone at 75°C.
Til en omrørt løsning av 2,537 g (2,0 mmol, regnet som 6'-N-Cbz-kana A (silyl)g) poly-(trimetylsilyl)-6'-N-Cbz-kana A fremstilt som i eksempel 1, i 32 ml sikttørket dietylketon ved 7 5°C ble det tilsatt en løsning av NAE (10,8 ml med 0,1944 mmol/ml dietylketon, 2,10 mmol) i løpet av 15 minutter. Omrøring ble fortsatt ved 7 5°C i ytterligere 3 timer, hvoretter blandingen ble helt i 150 ml vann. pH-verdien ble innstilt på 2,8 med 3N HCl, og dietylketonen ble avdampet under vakuum ved 4 0°C. HPLC-analyse av en aliquot av den vandige fase viste et utbytte på 39,18% di-Cbz-amikacin. To a stirred solution of 2.537 g (2.0 mmol, calculated as 6'-N-Cbz-cana A (silyl)g) poly-(trimethylsilyl)-6'-N-Cbz-cana A prepared as in Example 1, in 32 ml of sieve-dried diethyl ketone at 75°C, a solution of NAE (10.8 ml with 0.1944 mmol/ml diethyl ketone, 2.10 mmol) was added over 15 minutes. Stirring was continued at 75°C for a further 3 hours, after which the mixture was poured into 150 ml of water. The pH was adjusted to 2.8 with 3N HCl, and the diethyl ketone was evaporated under vacuum at 40°C. HPLC analysis of an aliquot of the aqueous phase showed a yield of 39.18% di-Cbz-amikacin.
Hovedparten av den vandige fase ble redusert under 3,39 atm. H2-trykk i 3 1/4 time ved 23°C under anvendelse av 1,0 g Pd/C katalysator. Katalysatoren ble fjernet ved filtrering, og kombinert filtrat og vaskevæsker ble analysert mikrobiologisk for amikacin. Turbidimetrisk analyse viste et utbytte på 27,84%, og plateanalyse viste 28,6% utbytte. The majority of the aqueous phase was reduced below 3.39 atm. H 2 pressure for 3 1/4 hours at 23°C using 1.0 g of Pd/C catalyst. The catalyst was removed by filtration, and the combined filtrate and washings were analyzed microbiologically for amikacin. Turbidimetric analysis showed a yield of 27.84%, and plate analysis showed a 28.6% yield.
Eksempel 15 Example 15
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl) -kana A med NAE ved 0-5°C etter tilbakehydrolysering med vann. Preparation of amikacin by acylation of poly-(trimethylsilyl) cana A with NAE at 0-5°C after back-hydrolysis with water.
A. Silylering av kanamycin A under anvendelse av HMDS med A. Silylation of kanamycin A using HMDS with
TMCS som katalysator. TMCS as a catalyst.
10 g kanamycin A med renhet 97,6% (20,14 mmol) i 100 ml sikttørket acetonitril ble brakt til å koke med tilbakekjøling under nitrogenatmosfære. En blanding av 22,76 g (141 mmol, 7 mol pr. mol kanamycin A) av HMDS og 1 ml (0,856 g, 7,88 mmol) TMCS ble tilsatt til den tilbakekjølte reaksjonsblanding i løpet av 10 minutter. Tilbakekjøling ble fortsatt i 4 3/4 time hvoretter blandingen ble avkjølt og konsentrert under vakuum til en gul, viskøs væske og tørket under høyvakuum i 2 timer. Utbyttet av produkt var 23,8 g (97,9%, regnet som kanamycin A (silyl)1Q). 10 g of kanamycin A with a purity of 97.6% (20.14 mmol) in 100 ml of sieve-dried acetonitrile was brought to boiling with reflux under a nitrogen atmosphere. A mixture of 22.76 g (141 mmol, 7 mol per mol kanamycin A) of HMDS and 1 mL (0.856 g, 7.88 mmol) of TMCS was added to the cooled reaction mixture over 10 minutes. Refrigeration was continued for 4 3/4 hours after which the mixture was cooled and concentrated under vacuum to a yellow viscous liquid and dried under high vacuum for 2 hours. The yield of product was 23.8 g (97.9%, calculated as kanamycin A (silyl)1Q).
B. Acylering B. Acylation
23,8 g (20,14 mmol) poly-(trimetylsilyl)-kanamycin A fremstilt under trinn A ovenfor ble løst i 250 ml sikttørket aceton ved 23°C og deretter avkjølt til 0-5°C. 3,63 ml vann (201,4 mmol, 10 mol pr. mol polysilylert kanamycin A) ble tilsatt under om-røring, og blandingen ble anbrakt under moderat vakuum i 30 minutter. NAE (19,133 mmol, 0,95 mol pr. mol polysilylert kanamycin A) i 108,3 ml aceton ble deretter tilsatt i løpet av mindre enn 1 minutt. Blandingen ble omrørt ved 0-5°C i 1 time, 23.8 g (20.14 mmol) of poly-(trimethylsilyl)-kanamycin A prepared during step A above was dissolved in 250 ml sieve-dried acetone at 23°C and then cooled to 0-5°C. 3.63 mL of water (201.4 mmol, 10 mol per mol of polysilylated kanamycin A) was added with stirring, and the mixture was placed under moderate vacuum for 30 minutes. NAE (19.133 mmol, 0.95 mol per mol polysilylated kanamycin A) in 108.3 mL of acetone was then added in less than 1 minute. The mixture was stirred at 0-5°C for 1 hour,
tynnet med vann, pH ble innstilt på 2,5 og acetonen deretter fjernet under vakuum. Den vandige løsning ble deretter redusert ved 3,4 0 atm. H2~trykk ved 23°C i 1\ time under anvendelse av 2,0 g 10% Pd/C som katalysator. Den reduserte reaksjonsblanding ble filtrert gjennom "Dicalite", konsentrert til ca. 100 ml under vakuum ved 40°C og deretter helt i en %CG-50" (NH4<+>) diluted with water, the pH was adjusted to 2.5 and the acetone then removed under vacuum. The aqueous solution was then reduced at 3.40 atm. H 2 -pressure at 23°C for 1/h using 2.0 g of 10% Pd/C as catalyst. The reduced reaction mixture was filtered through "Dicalite", concentrated to ca. 100 ml under vacuum at 40°C and then poured into a %CG-50" (NH4<+>)
kolonne (6 liter harpiks, 5 x 100 cm). Den ble vasket med vann og deretter eluert med 0,6N-1,0N-3N NH^OH. Det ble oppnådd 60,25% amikacin, 4,37% BB-K6, 4,35% BB-K29, 26,47% kanamycin A og 2,18% polyacylforbindelser. column (6 liters of resin, 5 x 100 cm). It was washed with water and then eluted with 0.6N-1.0N-3N NH^OH. 60.25% amikacin, 4.37% BB-K6, 4.35% BB-K29, 26.47% kanamycin A and 2.18% polyacyl compounds were obtained.
Eksempel 16 Example 16
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl) -6 ' -N-Cbz-kana A med SAE ved 0-5°C etter tilbake-metanolyse. Preparation of amikacin by acylation of poly-(trimethylsilyl)-6'-N-Cbz-cana A with SAE at 0-5°C after back methanolysis.
A. Silylering av 6'- N- Cbz- kanamycin A. A. Silylation of 6'-N-Cbz-kanamycin A.
20.0 g (32,4 mmol) 6'-N-Cbz-kanamycin A i 200 ml sikttørket acetonitril ble brakt til å koke med tilbakekjøling under nitrogenatmosfære. 47,3 ml HMDS (226,8 mmol, 7 mol pr. mol 6<1->N-Cbz-kana A) ble tilsatt i løpet av 10 minutter, og koking med tilbake-kjøling ble fortsatt i 20 timer. Blandingen ble deretter avkjølt, konsentrert under vakuum og tørket under høyvakuum i 2 timer, hvorved det ble oppnådd 39,1 g hvitt, amorft fast stoff (95,4% utbytte, regnet som 6<1->N-Cbz-kana A (silyl)g). 20.0 g (32.4 mmol) of 6'-N-Cbz-kanamycin A in 200 ml of sieve-dried acetonitrile was brought to boiling with reflux under a nitrogen atmosphere. 47.3 mL of HMDS (226.8 mmol, 7 mol per mol 6<1->N-Cbz-can A) was added over 10 minutes, and refluxing was continued for 20 hours. The mixture was then cooled, concentrated under vacuum and dried under high vacuum for 2 hours to give 39.1 g of white, amorphous solid (95.4% yield, calculated as 6<1->N-Cbz-kana A ( silyl)g).
B. Acylering. B. Acylation.
39.1 g (32,4 mmol) poly-(trimetylsilyl)-6'-N-Cbz-kana A fremstilt under trinn A ovenfor ble løst i 4 00 ml tørr aceton under omrøring ved 23°C. 6,6 ml metanol (162 mmol, 5 mol pr. mol polysilylert 6'-N-Cbz-kana A) ble tilsatt, og blandingen ble om- 39.1 g (32.4 mmol) of poly-(trimethylsilyl)-6'-N-Cbz-cana A prepared during step A above was dissolved in 400 ml of dry acetone with stirring at 23°C. 6.6 ml of methanol (162 mmol, 5 mol per mol polysilylated 6'-N-Cbz-cana A) was added, and the mixture was re-
rørt ved 23°C i 1 time under en sterk nitrogenstrøm. Blandingen ble avkjølt til 0-5°C, og en løsning av 11,35 g (32,4 mmol) SAE i 120 ml tørr aceton som var avkjølt på forhånd ble tilsatt. Blandingen ble omrørt i ytterligere 3 timer ved 0-5°C og deretter anbrakt i et 4°C kaldt rom i 1 uke. 3 00 ml vann ble tilsatt og pH innstilt på 2,0. Blandingen ble omrørt i 1 time og acetonen deretter avdampet under vakuum. Den resulterende vandige løsning ble redusert ved 3,67 atm. t^-trykk i 17 timer ved 23°C under anvendelse av 3,0 g 10% Pd/C som katalysator. Den ble deretter filtrert gjennom "Dicalite", konsentrert under vakuum til 75-100 ml og fyllt i en "CG-50" NH4<+>) kolonne og eluert med vann og 0,6N NH4OH. Det ble oppnådd 52,52% amikacin, 14,5% BB-K29, 19,6% kanamycin A og 1,71% polyacylforbindelser. stirred at 23°C for 1 hour under a strong stream of nitrogen. The mixture was cooled to 0-5°C and a solution of 11.35 g (32.4 mmol) of SAE in 120 ml of dry acetone which had been previously cooled was added. The mixture was stirred for a further 3 hours at 0-5°C and then placed in a 4°C cold room for 1 week. 300 ml of water was added and the pH adjusted to 2.0. The mixture was stirred for 1 hour and the acetone was then evaporated under vacuum. The resulting aqueous solution was reduced at 3.67 atm. t^ pressure for 17 hours at 23°C using 3.0 g of 10% Pd/C as catalyst. It was then filtered through "Dicalite", concentrated under vacuum to 75-100 ml and loaded onto a "CG-50" NH4<+>) column and eluted with water and 0.6N NH4OH. 52.52% amikacin, 14.5% BB-K29, 19.6% kanamycin A and 1.71% polyacyl compounds were obtained.
Eksempel 17 Example 17
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl) -kana A med SAE ved 0-5°C etter tilbakehydrolysering med vann. Preparation of amikacin by acylation of poly-(trimethylsilyl) cana A with SAE at 0-5°C after back-hydrolysis with water.
A. Silylering av kanamycin A med TMCS i acetonitril under A. Silylation of kanamycin A with TMCS in acetonitrile below
anvendelse av tetrametylguanidin som syreakseptor. use of tetramethylguanidine as acid acceptor.
4,88 g (10,07 mmol) kanamycin A ble suspendert i 100 ml sikttørket acetonitril under omrøring ved 23°C. Til den omrørte suspensjon ble det tilsatt 16,234 g (140,98 mmol, 14 mol pr. mol kanamycin A) tetrametylguanidin (TMG). Blandingen ble oppvarmet til tilbakekjøling, og 15,32 g (140,98 mmol, 14 mol pr. mol kanamycin A) TMCS ble tilsatt i løpet av 15 minutter. Det ble dannet et hvitt bunnfall av TMG*HCl etter at ca. halvparten av TMCS var blitt tilsatt. Blandingen ble avkjølt til romtemperatur, konsentrert til en klebrig rest og tørket under høyvakuum i 2 timer. 4.88 g (10.07 mmol) of kanamycin A was suspended in 100 ml of sieve-dried acetonitrile with stirring at 23°C. To the stirred suspension was added 16.234 g (140.98 mmol, 14 mol per mol kanamycin A) tetramethylguanidine (TMG). The mixture was heated to reflux and 15.32 g (140.98 mmol, 14 mol per mol kanamycin A) of TMCS was added over 15 minutes. A white precipitate of TMG*HCl was formed after approx. half of the TMCS had been added. The mixture was cooled to room temperature, concentrated to a sticky residue and dried under high vacuum for 2 hours.
Det faste stoff ble blandet grundig med 100 ml tørr THF, og det uløselige TMG*HCl ble frafiltrert og vasket med 5 x 20 ml porsjoner THF. Kombinert filtrat og vaskevæsker ble konsentrert under vakuum ved 4 0°C til en klebrig rest og tørket under høy-vakuum i 2 timer. Det ble oppnådd 10,64 g av en lys fløtefarget, klebrig rest (87,6% utbytte, regnet som kanamycin A (silyl)1Q)• The solid was mixed thoroughly with 100 ml of dry THF, and the insoluble TMG*HCl was filtered off and washed with 5 x 20 ml portions of THF. Combined filtrate and washings were concentrated under vacuum at 40°C to a sticky residue and dried under high vacuum for 2 hours. 10.64 g of a light cream-colored, sticky residue was obtained (87.6% yield, calculated as kanamycin A (silyl)1Q)•
B. Acylering B. Acylation
10,64 g (10,07 mmol) poly-(trimetylsilyl)-kanamycin A fremstilt under trinn A ovenfor ble løst i 110 ml sikttørket aceton under omrøring ved 23°C, og løsningen ble avkjølt til 0-5°C. Det ble tilsatt 1,81 ml (100,7 mmol, 10 mol pr. mol polysilylert kana A) vann, og løsningen ble omrørt i 3 0 minutter 10.64 g (10.07 mmol) of poly-(trimethylsilyl)-kanamycin A prepared during step A above was dissolved in 110 ml sieve-dried acetone with stirring at 23°C, and the solution was cooled to 0-5°C. 1.81 mL (100.7 mmol, 10 mol per mol polysilylated cana A) of water was added and the solution was stirred for 30 minutes
I v/ T U / I I w/ T U / I
under moderat vakuum. 3,70 g (10,57 mmol, 5% overskudd) SAE i 40 ml aceton som var avkjølt på forhånd ble tilsatt i løpet av mindre enn 1 minutt, og blandingen ble omrørt i 1 time. Blandingen ble opparbeidet ved den generelle fremgangsmåte i eksempel 16B, hvorved det ble oppnådd ca. 50% amikacin, ca. 10% BB-K29, 5-8% BB-K6, ca. 20% kanamycin A og 5-8% polyacylforbindelser. under moderate vacuum. 3.70 g (10.57 mmol, 5% excess) of SAE in 40 mL of pre-cooled acetone was added in less than 1 minute and the mixture was stirred for 1 hour. The mixture was worked up by the general method in example 16B, whereby approx. 50% amikacin, approx. 10% BB-K29, 5-8% BB-K6, approx. 20% kanamycin A and 5-8% polyacyl compounds.
Eksempel 18 Example 18
Fremstilling av poly-(trimetylsilyl)-kanamycin A i pyridin under anvendelse av HMDS. Preparation of poly-(trimethylsilyl)-kanamycin A in pyridine using HMDS.
10,0 g (20,64 mmol) kanamycin A ble suspendert i 100 ml sikttørket, nydestillert pyridin ved 23°C. Nitrogengjennomstrøm-ning ble påbegynt, og suspensjonen ble brakt til å koke med til-bakekjøling. 17,33 g (107,32 mmol, 5,2 mol pr. mol kanamycin A) HMDS ble tilsatt i løpet av 10 minutter, og blandingen ble kokt med tilbakekjøling i 19 timer. Den ble deretter avkjølt til romtemperatur, konsentrert under vakuum til en lysegul-gylden viskøs væske og tørket under høyvakuum til et hvitt, amorft fast stoff. Det ble oppnådd 22,1 g (92,6% utbytte, regnet som kanamycin A (silyl)1Q). 10.0 g (20.64 mmol) of kanamycin A was suspended in 100 ml of sieve-dried, freshly distilled pyridine at 23°C. Nitrogen flow was started and the suspension was brought to boiling with reflux. 17.33 g (107.32 mmol, 5.2 mol per mol kanamycin A) of HMDS was added over 10 minutes, and the mixture was refluxed for 19 hours. It was then cooled to room temperature, concentrated under vacuum to a light yellow-golden viscous liquid and dried under high vacuum to a white, amorphous solid. 22.1 g were obtained (92.6% yield, calculated as kanamycin A (silyl)1Q).
Eksempel 19 Example 19
Fremstilling av poly- (trietylsilyl)-kanamycin A under anvendelse av trietylklorsilan med trietylamin som syreakseptor. Preparation of poly-(triethylsilyl)-kanamycin A using triethylchlorosilane with triethylamine as acid acceptor.
5,0 g kanamycin A av 97,6% renhet (10,07 mmol) ble suspendert i 100 ml sikttørket acetonitril ved 23°C. 33,8 ml, 24,5 g (241,7 mmol) trietylamin (TEA) ble tilsatt, og suspensjonen ble brakt til å koke med tilbakekjøling. En løsning av23,7 ml, 21,3 g (140,98 mmol) trikloretylsilan i 25 ml tørr acetonitril ble tilsatt i løpet av 20 minutter. Kokingen med tilbakekjøling ble fortsatt i ytterligere 7 timer, og blandingen ble avkjølt til romtemperatur hvoretter det ble utskilt lange, fine nåler av TEA-HCl. Blandingen ble hensatt ved romtemperatur i 16 timer, konsentrert under vakuum ved 4 0°C til et klebrig, fast stoff og tørket i 2 timer under høyvakuum til et dyporange, klebrig fast stoff. Det faste stoff ble blandet grundig med 100 ml tørr THF ved 23°C, og det uløselige TEA*HCl ble frafiltrert, vasket med 5 x 20 ml THF og tørket hvorved det ble oppnådd 16,0 g TEA*HCl. Kombinert filtrat og vaskevæsker ble konsentrert under vakuum til et fast stoff og tørket under høyvakuum i 2 timer. Det ble oppnådd 19,3 g poly-(trietylsilyl)-kanamycin A som en 5.0 g of kanamycin A of 97.6% purity (10.07 mmol) was suspended in 100 ml of sieve-dried acetonitrile at 23°C. 33.8 mL, 24.5 g (241.7 mmol) of triethylamine (TEA) was added and the suspension was brought to reflux. A solution of 23.7 mL, 21.3 g (140.98 mmol) of trichloroethylsilane in 25 mL of dry acetonitrile was added over 20 minutes. The reflux was continued for another 7 hours and the mixture was cooled to room temperature after which long fine needles of TEA-HCl separated. The mixture was allowed to stand at room temperature for 16 hours, concentrated under vacuum at 40°C to a sticky solid and dried for 2 hours under high vacuum to a deep orange sticky solid. The solid was mixed thoroughly with 100 ml of dry THF at 23°C, and the insoluble TEA*HCl was filtered off, washed with 5 x 20 ml of THF and dried to give 16.0 g of TEA*HCl. Combined filtrate and washings were concentrated under vacuum to a solid and dried under high vacuum for 2 hours. 19.3 g of poly-(triethylsilyl)-kanamycin A were obtained as a
dyporange, viskøs væske. deep orange, viscous liquid.
Eksempel 20 Example 20
Fremstilling av poly-(trimetylsilyl)-kanamycin A under anvendelse av bis- trimetylsilylurea. Preparation of poly-(trimethylsilyl)-kanamycin A using bis-trimethylsilylurea.
10,0 g kanamycin A av renhet 99,7% (20,58 mmol) ble suspendert i 200 ml sikttørket acetonitril under omrøring ved 23°C. 10.0 g of kanamycin A of purity 99.7% (20.58 mmol) was suspended in 200 ml of sieve-dried acetonitrile with stirring at 23°C.
Til suspensjonen ble det tilsatt 29,45 g (144,01 mmol, 7 mol pr. mol kanamycin) bis-trimetylsilylurea (BSU), og blandingen ble brakt til å koke med tilbakekjøling under nitrogenatmosfære. Kokingen med tilbakekjøling ble fortsatt i 17 timer, hvoretter reaksjonsblandingen ble avkjølt til romtemperatur. Et lite kvantum nærværende uløselig materiale ble fjernet ved filtrering, vasket med 3 x 10 ml porsjoner acetonitril og tørket (1,1381 g). IR-analyse viste at dette var BSU pluss et lite kvantum ureagert kanamycin A. Kombinert filtrat og vaskevæsker ble avkjølt til 4°C i 16 timer. Ytterligere fast stoff ble utskilt, fjernet som ovenfor (7,8 g) og viste seg ved IR-analyse å være BSU pluss urea. Det lysegule filtrat og vaskevæskene ble konsentrert under vakuum ved 40°C og tørket under høyvakuum, hvorved det ble frembrakt 27,0 g av et hvitt, fast stoff som dels var klebrig og dels var fine, nålliknende krystaller. Det faste stoff ble behandlet med 150 ml heptan ved 23°C. Den uløselige del ble fjernet ved filtrering, vasket med 2 x 50 ml porsjoner heptan og tørket, hvorved det ble frembrakt 6,0 g hvite nåler (viste seg ved IR-analyse å være BSU pluss urea). Kombinert filtrat og vaskevæsker ble konsentrert under vakuum ved 4 0°C og tørket under høyvakuum i 2 timer, hvorved det ble frembrakt 2 0,4 g hvite nåler hvis IR-spektrum var typisk for polysilylert kanamycin A. Beregninger viste at produktet i gjennomsnitt inneholdt 7,22 trimetylsilylgrupper. To the suspension was added 29.45 g (144.01 mmol, 7 mol per mol kanamycin) of bis-trimethylsilylurea (BSU), and the mixture was brought to reflux under a nitrogen atmosphere. The reflux was continued for 17 hours, after which the reaction mixture was cooled to room temperature. A small amount of insoluble material present was removed by filtration, washed with 3 x 10 mL portions of acetonitrile and dried (1.1381 g). IR analysis showed this to be BSU plus a small amount of unreacted kanamycin A. Combined filtrate and washings were cooled to 4°C for 16 hours. Additional solid separated, removed as above (7.8 g) and was shown by IR analysis to be BSU plus urea. The pale yellow filtrate and washings were concentrated under vacuum at 40°C and dried under high vacuum, whereby 27.0 g of a white solid was produced which was partly sticky and partly fine, needle-like crystals. The solid was treated with 150 ml of heptane at 23°C. The insoluble portion was removed by filtration, washed with 2 x 50 mL portions of heptane and dried to yield 6.0 g of white needles (shown by IR analysis to be BSU plus urea). Combined filtrate and washings were concentrated under vacuum at 40°C and dried under high vacuum for 2 hours, yielding 20.4 g of white needles whose IR spectrum was typical of polysilylated kanamycin A. Calculations showed that the product contained, on average, 7.22 trimethylsilyl groups.
Eksempel 21 Example 21
Fremstilling av amikacin ved acylering av per-(trimetylsilyl) -kanamycin A etter partiell desilylering med 1,3-butandiol. Preparation of amikacin by acylation of per-(trimethylsilyl)-kanamycin A after partial desilylation with 1,3-butanediol.
A. Fremstilling av per-( trimetylsilyl)- kanamycin A. A. Preparation of per-(trimethylsilyl)-kanamycin A.
10,0 g (20,639 mmol) kanamycin A ble suspendert i 100 ml sikttørket acetonitril under omrøring ved 23°C. Suspensjonen ble brakt til å koke med tilbakekjøling under en nitrogenstrøm, og 23,322 g HMDS (144,5 mmol, 7 mol pr. mol kanamycin A) ble 10.0 g (20.639 mmol) of kanamycin A was suspended in 100 ml of sieve-dried acetonitrile with stirring at 23°C. The suspension was brought to reflux under a stream of nitrogen, and 23.322 g of HMDS (144.5 mmol, 7 mol per mol kanamycin A) was
Eksempel 24 Example 24
Fremstilling av poly-(trimetylsilyl)-kanamycin A under anvendelse av HMDS med imidazol som katalysator. 11 g (22,7 mmol) kanamycin A og 100 mg imidazol ble oppvarmet til tilbakekjøling i 100 ml sikttørket acetonitril under en nitrogenstrøm. 18,48 g (114,5 mmol, 5 mol pr. mol kanamycin A) HMDS ble tilsatt i løpet av 3 0 minutter, og blandingen ble kokt med tilbakekjøling i 20 timer. Fullstendig oppløsning inntraff på ca. 2\ time. Løsningen ble avkjølt til 23°C, og løsnings-midlet ble fjernet under vakuum og etterlot 21,6 g poly-(trimetylsilyl) -kanamycin A som en skumaktig rest (93,1% utbytte regnet som kanamycin (silyl) -q). Preparation of poly-(trimethylsilyl)-kanamycin A using HMDS with imidazole as catalyst. 11 g (22.7 mmol) of kanamycin A and 100 mg of imidazole were heated to reflux in 100 ml of sieve-dried acetonitrile under a stream of nitrogen. 18.48 g (114.5 mmol, 5 mol per mol kanamycin A) of HMDS was added over 30 minutes and the mixture was refluxed for 20 hours. Complete resolution occurred in approx. 2\ hours. The solution was cooled to 23°C and the solvent was removed under vacuum leaving 21.6 g of poly-(trimethylsilyl)-kanamycin A as a foamy residue (93.1% yield calculated as kanamycin (silyl)-q).
Eksempel 25 Example 25
Fremstilling av 1-N-[l-(-)-y-amino-a-hydroksybutyryl] - kanamycin B (BB-K26) ved acylering av poly-(trimetylsilyl)- kanamycin B med SAE. Preparation of 1-N-[l-(-)-γ-amino-α-hydroxybutyryl]-kanamycin B (BB-K26) by acylation of poly-(trimethylsilyl)-kanamycin B with SAE.
A. Fremstilling av poly-(trimetylsilyl)-kanamycin B under A. Preparation of poly-(trimethylsilyl)-kanamycin B below
anvendelse av HMDS med TMCS- katalysator. application of HMDS with TMCS catalyst.
25 g (51,7 mmol) kanamycin B i 250 ml sikttørket acetonitril ble oppvarmet til tilbakekjøling under en nitrogenstrøm. 62,3 g (385,81 mmol, 7,5 mol pr. mol kanamycin B) HMDS ble tilsatt i løpet av 3 0 minutter, etterfulgt av 1 ml TMCS som katalysator. Blandingen ble kokt med tilbakekjøling i 21 timer med fullstendig oppløsning etter 1 time. Løsningsmidlet ble fjernet under vakuum ved 60°C, og den oljeaktige rest ble holdt på 60°C under høy-vakuum i 3 timer. Det ble oppnådd 53,0 g poly-(trimetylsilyl)-kanamycin B (85,2% utbytte regnet som kanamycin B (silyl)1Q)• 25 g (51.7 mmol) of kanamycin B in 250 ml of sieve-dried acetonitrile was heated to reflux under a stream of nitrogen. 62.3 g (385.81 mmol, 7.5 mol per mol kanamycin B) of HMDS was added over 30 minutes, followed by 1 mL of TMCS as a catalyst. The mixture was refluxed for 21 hours with complete dissolution after 1 hour. The solvent was removed under vacuum at 60°C, and the oily residue was kept at 60°C under high vacuum for 3 hours. 53.0 g of poly-(trimethylsilyl)-kanamycin B were obtained (85.2% yield calculated as kanamycin B (silyl)1Q)•
B. Acylering B. Acylation
53,0 g av det i trinn A ovenfor fremstilte poly-(trimetylsilyl) -kanamycin B ble løst i 500 ml tørr aceton ved 0-5°C, 20,9 ml metanol ble tilsatt, og blandingen ble omrørt under vakuum i 30 minutter ved 0-5°C. En løsning av 18,1 g (51,67 mmol) SAE 53.0 g of the poly-(trimethylsilyl)-kanamycin B prepared in step A above was dissolved in 500 ml of dry acetone at 0-5°C, 20.9 ml of methanol was added, and the mixture was stirred under vacuum for 30 minutes at 0-5°C. A solution of 18.1 g (51.67 mmol) SAE
i 200 ml aceton som var blitt avkjølt på forhånd, ble tilsatt i løpet av mindre enn 1 minutt, og blandingen ble omrørt i 30 minutter ved 0-5°C. Blandingen ble opparbeidet i overensstemmelse med åen generelle fremgangsmåte i eksempel 22 og ble deretter fyllt i en kolonne av "CG-50" (NH4<+>) (8 x 120 cm). Den ble eluert med en NH^OH-gradient på fra 0,6N til 3N. Det ble oppnådd 38% BB-K26, 5% av det tilsvarende 6'-N-acylerte kanamycin B (BB-K22), 10% av det tilsvarende 3-N-acylerte kanamycin B (BB-K46), in 200 ml of pre-cooled acetone was added in less than 1 minute and the mixture was stirred for 30 minutes at 0-5°C. The mixture was worked up in accordance with the general procedure of Example 22 and was then loaded into a column of "CG-50" (NH4<+>) (8 x 120 cm). It was eluted with an NH^OH gradient from 0.6N to 3N. 38% BB-K26 was obtained, 5% of the corresponding 6'-N-acylated kanamycin B (BB-K22), 10% of the corresponding 3-N-acylated kanamycin B (BB-K46),
14,63% kanamycin B samt et lite kvantum polyacylert kanamycin B. 14.63% kanamycin B as well as a small amount of polyacylated kanamycin B.
Eksempel 26 Example 26
Fremstilling av poly-(trimetylsilyl)-kanamycin A under anvendelse av HMDS med kanamycin A sulfat som katalysator. 19,5 g (4 0,24 6 mmol) kanamycin A og 0,5 g (0,8 58 mmol) kanamycin A sulfat (totalt = 20,0 g, 41,0 mmol) i 200 ml sikt-tørket acetonitril ble brakt til koking med tilbakekjøling. 60,3 Preparation of poly-(trimethylsilyl)-kanamycin A using HMDS with kanamycin A sulfate as catalyst. 19.5 g (4 0.24 6 mmol) kanamycin A and 0.5 g (0.8 58 mmol) kanamycin A sulfate (total = 20.0 g, 41.0 mmol) in 200 ml sieve-dried acetonitrile were brought to a boil with reflux. 60.3
ml (287,7 mmol, 7 mol pr. mol kanamycin A) HMDS ble langsomt tilsatt, og blandingen ble kokt med tilbakekjøling i 28 timer. ml (287.7 mmol, 7 mol per mol kanamycin A) of HMDS was slowly added and the mixture was refluxed for 28 hours.
Den ble deretter inndampet til tørr tilstand i en rotasjonsfordamper og tørket under dampinjektorvakuum. Det ble oppnådd 47,5 g —poiy- (-trr-imetr<y>-l-s-i-l-<y>-l-)--kanamye-ln—A—som—en—blek<g>ul—oLj e—(-95-,-82% utbytte regnet som kanamycin A (silyl)1Q)• It was then evaporated to dryness in a rotary evaporator and dried under steam injector vacuum. 47.5 g of —poiy- (-trr-imetr<y>-l-s-i-l-<y>-l-)--kanamye-ln—A—as—a—pale<g>ul—oLj e—( -95-,-82% yield calculated as kanamycin A (silyl)1Q)•
Eksempel 27 Example 27
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl) -kanamycin A med N-trifluoracetylblokkert AHBA-N-hydroksysucc inimidester. Preparation of amikacin by acylation of poly-(trimethylsilyl)-kanamycin A with N-trifluoroacetyl blocked AHBA-N-hydroxysuccinimide ester.
A. Fremstilling av N-trifluoracetyl-AHBA og omdannelse til A. Preparation of N-trifluoroacetyl-AHBA and conversion to
dens N- hydroksysuccinimidester. its N-hydroxysuccinimide ester.
Til en suspensjon av 5,0 g (42 mmol) AHBA i 100 ml THF ble det tilsatt 40 g (191 mmol) trifluoreddiksyreanhydrid under om-røring i løpet av 10 minutter. Løsningen ble omrørt i 18 timer ved 23°C og deretter konsentrert til tørr tilstand under vakuum ved 50°C. Resten ble løst i 100 ml vandig metanol (1:1) og-om- To a suspension of 5.0 g (42 mmol) of AHBA in 100 ml of THF, 40 g (191 mmol) of trifluoroacetic anhydride was added with stirring during 10 minutes. The solution was stirred for 18 hours at 23°C and then concentrated to dryness under vacuum at 50°C. The residue was dissolved in 100 ml of aqueous methanol (1:1) and
rørt i 1 time. Den ble deretter konsentrert til tørr tilstand under vakuum og gjenoppløst i 50 ml H2O. Den vandige løsning ble ekstrahert med 3 x 50 ml porsjoner MIBK, og ekstrakten ble etter tørking over Na2S04 konsentrert til en olje. Spor av løs-ningsmiddel ble fjernet ved tilsetning og avdestillering av 4 ml vann. Ved oppbevaring forandret oljen seg til et voksaktig, krystallinsk fast stoff (2,5 g, 28% utbytte). stirred for 1 hour. It was then concentrated to dryness under vacuum and redissolved in 50 mL H2O. The aqueous solution was extracted with 3 x 50 ml portions of MIBK, and the extract was concentrated to an oil after drying over Na 2 SO 4 . Traces of solvent were removed by adding and distilling off 4 ml of water. On storage, the oil changed to a waxy, crystalline solid (2.5 g, 28% yield).
2,4 g (11,3 mmol) N-trifluoracetyl-AHBA ble løst i 50 ml 2.4 g (11.3 mmol) of N-trifluoroacetyl-AHBA were dissolved in 50 ml
tørr aceton, og 1,30 g (11,31 mmol) N-hydroksysuccinimid ble tilsatt til løsningen. En løsning av 2,33 g dicykloheksylkarbodiimid i 20 ml tørr aceton ble langsomt tilsatt. Reaksjonsblandingen ble omrørt i 2 timer ved 23°C, og det utfelte dicykloheksylurea ble fjernet ved filtrering og vasket med litt aceton. Kombinert filtrat og vaskevæsker (en løsning av N-hydroksysuccinimidesteren av N-trifluoracetyl-AHBA) ble anvendt i det neste trinn uten isolering. dry acetone, and 1.30 g (11.31 mmol) of N-hydroxysuccinimide was added to the solution. A solution of 2.33 g of dicyclohexylcarbodiimide in 20 ml of dry acetone was slowly added. The reaction mixture was stirred for 2 hours at 23°C, and the precipitated dicyclohexylurea was removed by filtration and washed with a little acetone. Combined filtrate and washings (a solution of the N-hydroxysuccinimide ester of N-trifluoroacetyl-AHBA) were used in the next step without isolation.
B. Acylering B. Acylation
Til en løsning av poly-(trimetylsilyl)-kanamycin A fremstilt som i eksempel 26 (11,31 mmol) i 54 ml aceton ble det tilsatt 2,0 ml (113,4 mmol) vann, og blandingen ble omrørt under vakuum ved 0-5°C i 30 minutter. N-hydroksysuccinimidesteren av N-trifluoracetyl-AHBA fremstilt i trinn A ovenfor (11,31 mmol) ble tilsatt til blandingen som deretter ble holdt på 5°C i 1 time. Deretter ble pH-verdien innstilt på ca. 2,0 med 20 prosentig I^SO^, blandingen omrørt i 3 0 minutter og pH-verdien deretter hevet til ca. 6,0 med NH^OH. Blandingen ble deretter inndampet i til tørr tilstand i en rotasjonsfordamper, hvorved det ble oppnådd 14,4 g av et klebrig, ikke helt hvitt, fast stoff. Det faste stoff ble løst i 100 ml vann, pH-verdien ble hevet fra 5,5 til 11,0 med ION NH^OH, og løsningen ble oppvarmet i et olje-bad ved 70°C i 1 time. pH (9,5) ble senket til 7,0 med HCl hvoretter løsningen ble finfiltrert for fjerning av et lite kvantum uløselige bestanddeler, og filtratet ble vasket med vann. Kombinert filtrat og vaskevæsker (188 ml) ble tilført til en "CG-50" (NH4<+>) kolonne (8 x 90 cm), vasket med 2 liter vann og eluert med en NH4OH-gradient (0,6N-1,ON-konsentrert). Det ble oppnådd 28,9% amikacin, 5,0% BB-K6, 5,7% BB-K29, 43,8% kanamycin A, 3,25% polyacylforbindelser samt 14,3% av et ukjent materiale som var i den første fraksjon fra kolonnen. To a solution of poly-(trimethylsilyl)-kanamycin A prepared as in Example 26 (11.31 mmol) in 54 ml of acetone was added 2.0 ml (113.4 mmol) of water, and the mixture was stirred under vacuum at 0 -5°C for 30 minutes. The N-hydroxysuccinimide ester of N-trifluoroacetyl-AHBA prepared in step A above (11.31 mmol) was added to the mixture which was then kept at 5°C for 1 hour. The pH value was then set to approx. 2.0 with 20 percent I^SO^, the mixture stirred for 30 minutes and the pH then raised to approx. 6.0 with NH^OH. The mixture was then evaporated to dryness in a rotary evaporator, whereby 14.4 g of a sticky, off-white, solid was obtained. The solid was dissolved in 100 ml of water, the pH was raised from 5.5 to 11.0 with ION NH 2 OH, and the solution was heated in an oil bath at 70°C for 1 hour. The pH (9.5) was lowered to 7.0 with HCl after which the solution was finely filtered to remove a small amount of insoluble constituents, and the filtrate was washed with water. Combined filtrate and washings (188 mL) were fed to a "CG-50" (NH4<+>) column (8 x 90 cm), washed with 2 liters of water and eluted with an NH4OH gradient (0.6N-1, ON-concentrated). 28.9% amikacin, 5.0% BB-K6, 5.7% BB-K29, 43.8% kanamycin A, 3.25% polyacyl compounds and 14.3% of an unknown material that was in the first fraction from the column.
Eksempel 28 Example 28
Fremstilling av amikacin ved acylering av poly-(trimetylsilyl) -kanamycin A med tert-butyloksykarbonyl-blokkert AHBA- N- hydroksysuccinimidester. Preparation of amikacin by acylation of poly-(trimethylsilyl)-kanamycin A with tert-butyloxycarbonyl-blocked AHBA-N-hydroxysuccinimide ester.
A. Fremstilling av tert-BOC-AHBA og omdannelse til dens N-hydroksysuccinimidester A. Preparation of tert-BOC-AHBA and conversion to its N-hydroxysuccinimide ester
En løsning av 5,0 g (4 2 mmol) AHBA i 100 ml vann og 20 ml aceton ble innstilt på pH 10 med 10N NaOH. I løpet av 3-4 minutter ble det tilsatt 11,6 g (53 mmol) di-tert-butyldikarbonat, og løsningen ble omrørt i 35 minutter, idet pH-verdien ble holdt på 10 ved periodisk tilsetning av 10N NaOH. Acetonen ble fjernet under vakuum, og den vandige fase ble vasket med 4 0 ml etylacetat. pH-verdien til den vandige løsning ble senket til 2,0 med 3N HCl, og løsningen ble deretter ekstrahert med 3 x 3 0 ml MIBK. De kombinerte MIBK-ekstraker ble tørket over Na2S04 og konsentrert til en klar, oljeaktig rest (8,2 g, 89%). A solution of 5.0 g (42 mmol) AHBA in 100 ml water and 20 ml acetone was adjusted to pH 10 with 10N NaOH. In the course of 3-4 minutes, 11.6 g (53 mmol) of di-tert-butyl dicarbonate were added, and the solution was stirred for 35 minutes, the pH value being maintained at 10 by periodic addition of 10N NaOH. The acetone was removed under vacuum and the aqueous phase was washed with 40 ml of ethyl acetate. The pH of the aqueous solution was lowered to 2.0 with 3N HCl, and the solution was then extracted with 3 x 30 mL of MIBK. The combined MIBK extracts were dried over Na 2 SO 4 and concentrated to a clear oily residue (8.2 g, 89%).
4,25 g (19,4 mmol) tert-BOC-AHBA ble løst i 50 ml aceton, 4.25 g (19.4 mmol) of tert-BOC-AHBA was dissolved in 50 ml of acetone,
og det ble tilsatt 2,23 g (19,4 mmol) N-hydroksysuccinimid. En løsning av 4,00 g (19,4 mmol) dicykloheksylkarbodiimid i 20 ml aceton ble langsomt tilsatt, og blandingen ble omrørt i 2 timer ved 23°C. Det utfelte dicykloheksylurea ble fjernet ved filtrering og vasket med et lite kvantum aceton. Kombinert filtrat og vaskevæsker (en løsning av N-hydroksysuccinimidesteren av tert-BOC-AHBA) ble anvendt i neste trinn uten isolering. and 2.23 g (19.4 mmol) of N-hydroxysuccinimide was added. A solution of 4.00 g (19.4 mmol) of dicyclohexylcarbodiimide in 20 ml of acetone was slowly added, and the mixture was stirred for 2 hours at 23°C. The precipitated dicyclohexylurea was removed by filtration and washed with a small amount of acetone. Combined filtrate and washings (a solution of the N-hydroxysuccinimide ester of tert-BOC-AHBA) were used in the next step without isolation.
B. Acylering B. Acylation
Til en løsning av poly-(trimetylsilyl)-kanamycin A fremstilt som i eksempel 26 (41,28 mmol) i 94 ml aceton ble det tilsatt 3,5 ml (194 mmol) vann, og blandingen ble omrørt under vakuum ved 0-5°C i 3 0 minutter. N-hydroksysuccinimidesteren av tert-BOC-AHBA fremstilt i trinn A ovenfor (19,4 mmol) ble tilsatt, og blandingen ble hensatt ved 5°C i 1 time. Det ble tilsatt 2 00 ml vann og pH senket fra 7,0 til 2,0 med 2 0 prosentig H^ SO^. Etter 30 minutters omrøring ble pH hevet til ca. 6,0 med NH^OH, og blandingen ble inndampet til tørr tilstand under vakuum, hvorved det ble frembrakt 36,3 g av en gylden olje. Oljen ble løst i 200 ml trifluoreddiksyre, hensatt i 15 minutter og inndampet til tørr tilstand i en rotasjonsfordamper. Oljen ble vasket med vann, og vannet ble hurtigavdampet. Konsentrert NH^OH ble tilsatt til pH 6,0 og hurtigavdampet. Det resulterende faste stoff ble løst i vann og filtrert, og filteret ble vasket med vann. Kombinert filtrat og vaskevæsker (259 ml) ble fyllt i en "CG-50" (NH4<+>) kolonne (8 x 92 cm), vasket med 4 liter vann og eluert med en NH4OH-gradient (0", 6N-1, ON-konsentrert) . Det ble oppnådd 40,32% amikacin, 4,58% BB-K6, 8,32% BB-K29, 30,50% kanamycin A og 7,43% polyacylforbindelser. To a solution of poly-(trimethylsilyl)-kanamycin A prepared as in Example 26 (41.28 mmol) in 94 ml of acetone was added 3.5 ml (194 mmol) of water, and the mixture was stirred under vacuum at 0-5 °C for 30 minutes. The N-hydroxysuccinimide ester of tert-BOC-AHBA prepared in Step A above (19.4 mmol) was added and the mixture was allowed to stand at 5°C for 1 hour. 200 ml of water was added and the pH lowered from 7.0 to 2.0 with 20 percent H 2 SO 2 . After 30 minutes of stirring, the pH was raised to approx. 6.0 with NH 3 OH, and the mixture was evaporated to dryness under vacuum to give 36.3 g of a golden oil. The oil was dissolved in 200 ml of trifluoroacetic acid, left for 15 minutes and evaporated to dryness in a rotary evaporator. The oil was washed with water, and the water was rapidly evaporated. Concentrated NH^OH was added to pH 6.0 and rapidly evaporated. The resulting solid was dissolved in water and filtered, and the filter was washed with water. Combined filtrate and washings (259 mL) were loaded into a "CG-50" (NH4<+>) column (8 x 92 cm), washed with 4 liters of water and eluted with an NH4OH gradient (0", 6N-1 , ON-concentrated) 40.32% amikacin, 4.58% BB-K6, 8.32% BB-K29, 30.50% kanamycin A and 7.43% polyacyl compounds were obtained.
Eksempel 29 Example 29
Den generelle fremgangsmåte fra eksempel 1 ble gjentatt, bortsett fra at det der anvendte 6<1->N-karbobenzyloksykanamycin A ble erstattet med en ekvimolar vektmengde 6'-N-karbobenzyloksykanamycin B, og det ble derved fremstilt et 1-N-[l-(-)-y-amino-a-hydroksybutyryl] -kanamycin B. The general procedure from Example 1 was repeated, except that the 6<1->N-carbobenzyloxykanamycin A used there was replaced by an equimolar amount by weight of 6'-N-carbobenzyloxykanamycin B, and a 1-N-[l -(-)-γ-amino-α-hydroxybutyryl]-kanamycin B.
Eksempel 3 0 Example 3 0
Den generelle fremgangsmåte fra eksempel 1 ble gjentatt, bortsett fra at det istedenfor den der anvendte L-(-)-y-benzyl-oksykarbonylamino-a-hydroksysmørsyre-N-hydroksy-5-norbornen-2,3-dikarboksimidester ble erstattet med henholdsvis L-(-)-3-benzyl-oksykarbonylamino-a-hydroksypropionsyre-N-hydroksy-5-norbornen-2 , 3-dikarboksimidester og L- (-) -6 -benzy lok sy karbonyl arnino-a - hydroksyvaleriansyre-N-hydroksy-5-norbornen-2,3-dikarboksimidester, og det ble derved fremstilt henholdsvis l-N-[L-(-)-B-amino-a-hydroksypropiony1]-kanamycin A og l-N-[L-(-)-5-amino-a-hydroksyvaleryl]-kanamycin A. The general procedure from example 1 was repeated, except that instead of the L-(-)-γ-benzyl-oxycarbonylamino-α-hydroxybutyric acid-N-hydroxy-5-norbornene-2,3-dicarboximide ester used there, it was replaced with L-(-)-3-benzyl-oxycarbonylamino-α-hydroxypropionic acid-N-hydroxy-5-norbornene-2 , 3-dicarboximide ester and L-(-)-6-benzyl oxycarbonylamino-α-hydroxyvaleric acid-N- hydroxy-5-norbornene-2,3-dicarboximide ester, and 1-N-[L-(-)-B-amino-α-hydroxypropiony1]-kanamycin A and 1-N-[L-(-)-5- amino-α-hydroxyvaleryl]-kanamycin A.
Eksempel 31 Example 31
Den generelle fremgangsmåte fra eksempel 25 ble gjentatt, bortsett fra at det istedenfor den der anvendte L-(-)-Y-benzyl-oksykarbonylamino-a-hydroksysmørsyre-N-hydroksyester ble anvendt henholdsvis L-(-)-ø-benzyloksykarbonylamino-a-hydroksypropionsyre-N-hydroksysuccinimidester og L-(-)-6-benzyloksykarbonylamino-a-hydroksyvaleriansyre-N-hydroksysuccinimidester, og det ble derved fremstilt henholdsvis The general procedure from example 25 was repeated, except that instead of the L-(-)-Y-benzyl-oxycarbonylamino-α-hydroxybutyric acid-N-hydroxyester used there, respectively L-(-)-ø-benzyloxycarbonylamino-α -hydroxypropionic acid-N-hydroxysuccinimide ester and L-(-)-6-benzyloxycarbonylamino-α-hydroxyvaleric acid-N-hydroxysuccinimide ester, and it was thereby prepared respectively
l-N-[L-(-)-B-amino-a-hydroksypropionyl]-kanamycin B og l-N-[L-(-)-B-amino-α-hydroxypropionyl]-kanamycin B and
l-N-(L-(-)-if-amino-a-hydroksyvaleryl]-kanamycin B. l-N-(L-(-)-if-amino-α-hydroxyvaleryl]-kanamycin B.
Eksempel 32 Example 32
Fremstilling av BB-K8 ved acylering av poly-(trimetylsilyl)-3^ 6'- di- N- karbobenzyloksykanamycin A i vannfri dietylketon. Preparation of BB-K8 by acylation of poly-(trimethylsilyl)-3^6'-di-N-carbobenzyloxykanamycin A in anhydrous diethyl ketone.
A./ 3, 6'- di- N- karbobenzyloksykanamycin A. A./ 3, 6'- di- N- carbobenzyloxykanamycin A.
En suspensjon av 7,26 g (15 mmol) kanamycin A (fri base) og 18,6 g (75 mmol) nikkelacetattetrahydrat i 300 ml dimetylsulf-Qksyd (DMSO) ble holdt oppvarmet ved 100°C i ca. 30 minutter under omrøring, inntil det var oppnådd en klar, grønn løsning. Etter avkjøling ble det tilsatt en løsning av 11,8 g (37,6 mmol) N-karbobenzyloksy-5-norbornen-2,3-dikarboksimid i 50 ml DMSO A suspension of 7.26 g (15 mmol) kanamycin A (free base) and 18.6 g (75 mmol) nickel acetate tetrahydrate in 300 ml dimethylsulfoxide (DMSO) was kept heated at 100°C for approx. 30 minutes with stirring, until a clear, green solution was obtained. After cooling, a solution of 11.8 g (37.6 mmol) of N-carbobenzyloxy-5-norbornene-2,3-dicarboximide in 50 ml of DMSO was added
til løsningen. Blandingen ble omrørt ved romtemperatur natten over, behandlet med 100 ml kons. ammoniakkvann og 1 liter vann, omrørt ved romtemperatur i 1 time og overført til toppen av en "Diaion HP-10"-søyle (300 ml). Søylen ble underkastet trinnvis eluering, først med 7 n NH^OH, deretter med metanol-vann (1:1) og endelig med metanol-vann (10:1) under oppsamling av 20 ml fraksjoner og overvåkning ved hjelp av tynnsjiktskromatografi på "Merck-Silica Gel 60 F-254"-plater under anvendelse av kloroform-etanol-28% ammoniumhydroksyd (1:2:1). En porsjon av det ønskete produkt, som krystalliserte som fine. nåler fra fraksjoner som to the solution. The mixture was stirred at room temperature overnight, treated with 100 ml conc. ammonia water and 1 liter of water, stirred at room temperature for 1 hour and transferred to the top of a "Diaion HP-10" column (300 ml). The column was subjected to stepwise elution, first with 7 n NH^OH, then with methanol-water (1:1) and finally with methanol-water (10:1) collecting 20 ml fractions and monitoring by thin-layer chromatography on "Merck -Silica Gel 60 F-254" plates using chloroform-ethanol-28% ammonium hydroxide (1:2:1). A portion of the desired product, which crystallized as fine. needles from factions that
inneholdt produktet i høye konsentrasjoner, ble frafiltrert, hvorved man fikk en analytisk prøve. Filtratet og de andre fraksjoner som inneholdt det ønskete produkt (RF 0,42) ble kombinert, og løsningen ble inndampet under vakuum. Resten ble blandet grundig med dietyleter, hvormed man fikk totalt 9,7 g (86%) av det i overskriften anførte produkt. Smp. 300°C. IR(KBr): uc=o 1690 cm"<1>. NMR (DMSO-dg) + DC1, pD ca. 3): 6 4,76-5,26 (6^1^*1^ , 11^' og CO-OCH2-CgH5 x 2), 7,26 (10H, s, CO-OCH2-CfiH5 x 2). contained the product in high concentrations, was filtered off, whereby an analytical sample was obtained. The filtrate and the other fractions containing the desired product (RF 0.42) were combined, and the solution was evaporated under vacuum. The residue was thoroughly mixed with diethyl ether, with which a total of 9.7 g (86%) of the title product was obtained. Temp. 300°C. IR(KBr): uc=o 1690 cm"<1>. NMR (DMSO-dg) + DC1, pD ca. 3): 6 4.76-5.26 (6^1^*1^ , 11^' and CO-OCH2-CgH5 x 2), 7.26 (10H, s, CO-OCH2-CfH5 x 2).
Analyse: Beregnet for <C>34<H>48<N>4°15'H20: C: 52'98' H: 6»54» N: 7 >21 Analysis: Calculated for <C>34<H>48<N>4°15'H20: C: 52'98' H: 6»54» N: 7 >21
Funnet: C: 53,20, H: 6,42, *N: 7,04. Found: C: 53.20, H: 6.42, *N: 7.04.
B. Poly-( trimetylsilyl)- 3, 6'- di- N- karbobenzyloksykanamycin A B. Poly-(trimethylsilyl)-3,6'-di-N-carbobenzyloxykanamycin A
En blanding av 1,5 g (2 mmol) 3,6'-di-N-karbobenzyloksykanamycin A fra trinn A ovenfor og 1,29 g (8 mmol) heksametyldisilazan i 15 ml tørr acetonitril ble kokt med tilbakekjøling i 16 timer. Den klare løsning ble konsentrert til tørr tilstand under vakuum, og resten ble løst i 20 ml tørr dietylketon. Løs-ningen ble anvendt direkte i det neste trinn. A mixture of 1.5 g (2 mmol) of 3,6'-di-N-carbobenzyloxykanamycin A from Step A above and 1.29 g (8 mmol) of hexamethyldisilazane in 15 ml of dry acetonitrile was refluxed for 16 hours. The clear solution was concentrated to dryness under vacuum, and the residue was dissolved in 20 ml of dry diethyl ketone. The solution was used directly in the next step.
C. Acylering av poly-( trimetylsilyl)- 3, 6'- di- N- karbobenzyloksykanamycin A under anvendelse av en ekvimolar mengde acyleringsmiddel C. Acylation of poly-(trimethylsilyl)-3,6'-di-N-carbobenzyloxykanamycin A using an equimolar amount of acylating agent
Til løsningen fra trinn B ovenfor ble det under omrøring tilsatt 700 mg (2 mmol) L-(-)-a-karbobenzyloksyamino-a-hydroksy-smørsyre-N-hydroksysuccinimidester (SAE). Blandingen ble omrørt ved romtemperatur i 19 timer, deretter behandlet med 8 ml vann og 35 ml tetrahydrofuran (THF), innstilt på pH 3 med vandig saltsyre, omrørt i 30 minutter og konsentrert til tørr tilstand under vakuum. Resten ble løst i en blanding av 300 ml vann, 40 ml metanol, 10 ml n-butanol og 40 ml THF og hydrogenert natten over 1 nærvær av 500 mg 10% palladium-på-karbon. Katalysatoren ble fjernet ved filtrering, og filtratet ble inndampet under vakuum og lyofilisert, hvorved man fikk 1,7 g rå BB-K8. Det amorfe pulver ble gjenoppløst i vann, og løsningen ble innstilt på pH To the solution from step B above, 700 mg (2 mmol) of L-(-)-α-carbobenzyloxyamino-α-hydroxybutyric acid N-hydroxysuccinimide ester (SAE) was added with stirring. The mixture was stirred at room temperature for 19 hours, then treated with 8 mL of water and 35 mL of tetrahydrofuran (THF), adjusted to pH 3 with aqueous hydrochloric acid, stirred for 30 minutes, and concentrated to dryness under vacuum. The residue was dissolved in a mixture of 300 ml water, 40 ml methanol, 10 ml n-butanol and 40 ml THF and hydrogenated overnight in the presence of 500 mg 10% palladium-on-carbon. The catalyst was removed by filtration, and the filtrate was evaporated under vacuum and lyophilized to give 1.7 g of crude BB-K8. The amorphous powder was redissolved in water and the solution was adjusted to pH
4 med vandig saltsyre og ble kromatografert i en søyle av 4 with aqueous hydrochloric acid and was chromatographed in a column of
"Amberlite CG-50" i NH4<+->syklusen. Søylen ble underkastet trinnvis elusring med vann, 0,1 n NH^OH, 0,3 n NH^OH, 0,5 n NH^OH og 2 n NH^OH, under oppsamling av 10 ml fraksjoner og overvåkning ved hjelp av tynnsjiktskromatografi under anvendelse av "Merck "Amberlite CG-50" in the NH4<+->cycle. The column was subjected to stepwise elution with water, 0.1 n NH^OH, 0.3 n NH^OH, 0.5 n NH^OH and 2 n NH^OH, collecting 10 ml fractions and monitoring by thin layer chromatography using "Merck
Silica Gel 60 F-254"-plater under anvendelse av kloroform-metanol-28% ammoniumhydroksyd-vann (1:4:2:1). De homogene fraksjoner ble kombinert, inndampet og til slutt lyofilisert. Fraksjoner som inneholdt BB-K8 og fraksjoner som inneholdt gjenvunnet kanamycin A ble analysert under anvendelse av henholdsvis K. pneumonia A20680 og B. subtilis PCI 129. Silica Gel 60 F-254" plates using chloroform-methanol-28% ammonium hydroxide-water (1:4:2:1). The homogeneous fractions were combined, evaporated and finally lyophilized. Fractions containing BB-K8 and fractions containing recovered kanamycin A were analyzed using K. pneumonia A20680 and B. subtilis PCI 129, respectively.
Hydrogenolyse av det delvis avblokkerte produkt med Pd-c etterfulgt av isolering i en "CG-50"-søyle ga ytterligere 30 mg (2%) BB-K8. Det totale utbytte av BB-K8 ble 846 mg (69%). Hydrogenolysis of the partially deblocked product with Pd-c followed by isolation on a "CG-50" column gave an additional 30 mg (2%) of BB-K8. The total yield of BB-K8 was 846 mg (69%).
D. Acylering av poly-( trimetylsilyl- 3, 6'- di- N- karbobenzyloksykanamycin A under anvendelse av 1, 2 ekvivalenter acyleringsmidde1 D. Acylation of poly-(trimethylsilyl-3,6'-di-N-carbobenzyloxykanamycin A using 1.2 equivalents of acylating agent1
Trinn c ovenfor ble gjentatt, bortsett fra at det ble anvendt et 20% overskudd av acyleringsmiddel. Det ble oppnådd følgende resultater: Step c above was repeated, except that a 20% excess of acylating agent was used. The following results were obtained:
Hydrogenolyse av det delvis avblokkerte produkt med Pd-C etterfulgt av isolering i en "CG-50"-søyle ga ytterligere 21 mg (2%) BB-K8. Det totale utbytte av BB-K8 ble 771 mg (62%). Hydrogenolysis of the partially deblocked product with Pd-C followed by isolation on a "CG-50" column gave an additional 21 mg (2%) of BB-K8. The total yield of BB-K8 was 771 mg (62%).
E. Acylering av poly-( trimetylsilyl- 3, 6'- di- N- karbobenzyloksykanamycin A under anvendelse av 1, 5 ekvivalenter acyleringsmiddel E. Acylation of poly-(trimethylsilyl-3,6'-di-N-carbobenzyloxykanamycin A using 1.5 equivalents of acylating agent
Trinn C ovenfor ble gjentatt, bortsett fra at det ble anvendt et 50% overskudd av acyleringsmiddel. det ble oppnådd følgende resultater: Step C above was repeated, except that a 50% excess of acylating agent was used. the following results were obtained:
Claims (4)
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US79180677A | 1977-04-28 | 1977-04-28 | |
US88858578A | 1978-03-20 | 1978-03-20 |
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NO781437A NO149635C (en) | 1977-04-28 | 1978-04-25 | PROCEDURE FOR THE PREPARATION OF 1, N- (OMEGA-AMINO-ALFA-HYDROXYALKANOYL) -CANAMYCINE A OR B |
NO83831197A NO154697C (en) | 1977-04-28 | 1983-04-05 | DERIVATIVES OF CANAMYCIN A OR B. |
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NO781437A NO149635C (en) | 1977-04-28 | 1978-04-25 | PROCEDURE FOR THE PREPARATION OF 1, N- (OMEGA-AMINO-ALFA-HYDROXYALKANOYL) -CANAMYCINE A OR B |
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CA (2) | CA1105454A (en) |
CH (1) | CH639104A5 (en) |
CS (1) | CS207607B2 (en) |
CY (1) | CY1241A (en) |
DE (2) | DE2818992A1 (en) |
DK (2) | DK172543B1 (en) |
EG (1) | EG13710A (en) |
ES (1) | ES469303A1 (en) |
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NL (1) | NL178790C (en) |
NO (2) | NO149635C (en) |
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JPS55306A (en) * | 1978-04-28 | 1980-01-05 | Bristol Myers Co | Manufacture of 11nn*omegaaaminooalphaahydroxy alkanoyl**aminoglycoside antibiotic |
EP1484317A4 (en) * | 2002-02-19 | 2006-08-30 | Amato Pharm Prod Ltd | Lactic acid derivative |
EP3150617B1 (en) | 2007-11-21 | 2020-04-08 | Cipla USA, Inc. | Antibacterial aminoglycoside analogs |
WO2010132759A1 (en) | 2009-05-15 | 2010-11-18 | Achaogen, Inc. | Antibacterial derivatives of dibekacin |
WO2010132760A1 (en) | 2009-05-15 | 2010-11-18 | Achaogen, Inc. | Antibacterial derivatives of tobramycin |
WO2010132757A2 (en) | 2009-05-15 | 2010-11-18 | Achaogen, Inc. | Antibacterial aminoglycoside analogs |
WO2010132768A1 (en) | 2009-05-15 | 2010-11-18 | Achaogen, Inc. | Antibacterial derivatives of sisomicin |
WO2010132765A2 (en) | 2009-05-15 | 2010-11-18 | Achaogen, Inc. | Antibacterial aminoglycoside analogs |
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1978
- 1978-04-21 AR AR271852A patent/AR218910A1/en active
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- 1978-04-26 DK DK197801830A patent/DK172543B1/en not_active IP Right Cessation
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- 1978-04-27 IE IE841/78A patent/IE46945B1/en not_active IP Right Cessation
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