WO2002012267A1 - Procede de production de derives d'acide biliaire a fluoresceine - Google Patents
Procede de production de derives d'acide biliaire a fluoresceine Download PDFInfo
- Publication number
- WO2002012267A1 WO2002012267A1 PCT/GB2001/003559 GB0103559W WO0212267A1 WO 2002012267 A1 WO2002012267 A1 WO 2002012267A1 GB 0103559 W GB0103559 W GB 0103559W WO 0212267 A1 WO0212267 A1 WO 0212267A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- compound
- general formula
- formula
- salt
- alkali metal
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims abstract description 22
- 238000004519 manufacturing process Methods 0.000 title description 6
- -1 fluorescein bile acid derivatives Chemical class 0.000 title description 3
- 150000001875 compounds Chemical class 0.000 claims abstract description 36
- 229910052783 alkali metal Inorganic materials 0.000 claims abstract description 12
- 150000001340 alkali metals Chemical group 0.000 claims abstract description 12
- 150000003839 salts Chemical class 0.000 claims abstract description 12
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical class C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 claims abstract description 11
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims abstract description 11
- 239000011369 resultant mixture Substances 0.000 claims abstract description 6
- 239000003513 alkali Substances 0.000 claims abstract description 5
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 3
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 3
- 229910052760 oxygen Inorganic materials 0.000 claims abstract description 3
- 229910052700 potassium Inorganic materials 0.000 claims abstract description 3
- 238000004366 reverse phase liquid chromatography Methods 0.000 claims abstract description 3
- 229910052708 sodium Inorganic materials 0.000 claims abstract description 3
- 239000011734 sodium Substances 0.000 claims abstract description 3
- 229910052717 sulfur Inorganic materials 0.000 claims abstract description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims abstract 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims abstract 2
- 239000011591 potassium Substances 0.000 claims abstract 2
- 239000000243 solution Substances 0.000 claims description 20
- 238000006243 chemical reaction Methods 0.000 claims description 19
- 230000000903 blocking effect Effects 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- RIFGWPKJUGCATF-UHFFFAOYSA-N ethyl chloroformate Chemical compound CCOC(Cl)=O RIFGWPKJUGCATF-UHFFFAOYSA-N 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 2
- 239000012670 alkaline solution Substances 0.000 claims description 2
- 125000000896 monocarboxylic acid group Chemical group 0.000 claims description 2
- 239000012454 non-polar solvent Substances 0.000 claims description 2
- 230000020477 pH reduction Effects 0.000 claims description 2
- 150000003512 tertiary amines Chemical class 0.000 claims description 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 33
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 20
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 15
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 15
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 14
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- 239000004472 Lysine Substances 0.000 description 12
- 238000003756 stirring Methods 0.000 description 11
- 239000000377 silicon dioxide Substances 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 238000004128 high performance liquid chromatography Methods 0.000 description 8
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 7
- 229910052757 nitrogen Inorganic materials 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 6
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 239000011541 reaction mixture Substances 0.000 description 6
- ZUJFMZPBQQCXQR-UHFFFAOYSA-N 5-[[5-carboxy-5-[4-(3,7,12-trihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl)pentanoylamino]pentyl]carbamothioylamino]-2-(3-hydroxy-6-oxoxanthen-9-yl)benzoic acid Chemical compound OC1CC2CC(O)CCC2(C)C(CC(O)C23C)C1C3CCC2C(C)CCC(=O)NC(C(O)=O)CCCCNC(=S)NC1=CC=C(C2=C3C=CC(=O)C=C3OC3=CC(O)=CC=C32)C(C(O)=O)=C1 ZUJFMZPBQQCXQR-UHFFFAOYSA-N 0.000 description 5
- 239000003480 eluent Substances 0.000 description 5
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 5
- 239000002244 precipitate Substances 0.000 description 5
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 description 4
- FTOTVNJFGHCOCZ-UHFFFAOYSA-N Lysine conjugated cholic acid Chemical compound OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(=O)NC(CCCCN)C(O)=O)C)C1(C)C(O)C2 FTOTVNJFGHCOCZ-UHFFFAOYSA-N 0.000 description 4
- 239000003613 bile acid Substances 0.000 description 4
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000012044 organic layer Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 239000004380 Cholic acid Substances 0.000 description 3
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 3
- 229960002471 cholic acid Drugs 0.000 description 3
- 235000019416 cholic acid Nutrition 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- RUDATBOHQWOJDD-UHFFFAOYSA-N (3beta,5beta,7alpha)-3,7-Dihydroxycholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 RUDATBOHQWOJDD-UHFFFAOYSA-N 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- GKQLYSROISKDLL-UHFFFAOYSA-N EEDQ Chemical compound C1=CC=C2N(C(=O)OCC)C(OCC)C=CC2=C1 GKQLYSROISKDLL-UHFFFAOYSA-N 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-M Formate Chemical compound [O-]C=O BDAGIHXWWSANSR-UHFFFAOYSA-M 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 159000000000 sodium salts Chemical class 0.000 description 2
- 238000003828 vacuum filtration Methods 0.000 description 2
- DKPMWHFRUGMUKF-UHFFFAOYSA-N (3alpha,5alpha,6alpha,7alpha)-3,6,7-Trihydroxycholan-24-oic acid Natural products OC1C(O)C2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 DKPMWHFRUGMUKF-UHFFFAOYSA-N 0.000 description 1
- HULQGYPWEGNXPA-PBDHEXIJSA-N (4r)-4-[(8r,9s,10s,13r,14s,17r)-3-hydroxy-10,13-dimethyl-4,5,6,7,8,9,11,12,14,15,16,17-dodecahydro-1h-cyclopenta[a]phenanthren-17-yl]pentanoic acid Chemical compound C1CC2CC(O)=CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@@H](CCC(O)=O)C)[C@@]1(C)CC2 HULQGYPWEGNXPA-PBDHEXIJSA-N 0.000 description 1
- JOYGXTIHTHBSOA-UHFFFAOYSA-N 1-(4-chlorophenyl)-3-thiophen-2-ylprop-2-en-1-one Chemical compound C1=CC(Cl)=CC=C1C(=O)C=CC1=CC=CS1 JOYGXTIHTHBSOA-UHFFFAOYSA-N 0.000 description 1
- FCSKOFQQCWLGMV-UHFFFAOYSA-N 5-{5-[2-chloro-4-(4,5-dihydro-1,3-oxazol-2-yl)phenoxy]pentyl}-3-methylisoxazole Chemical compound O1N=C(C)C=C1CCCCCOC1=CC=C(C=2OCCN=2)C=C1Cl FCSKOFQQCWLGMV-UHFFFAOYSA-N 0.000 description 1
- RPKLZQLYODPWTM-NIRKWIOJSA-N 5alpha-cholanic acid Chemical compound C([C@@H]1CC2)CCC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 RPKLZQLYODPWTM-NIRKWIOJSA-N 0.000 description 1
- 0 CC(CC1=C(c(c(*)c2)ccc2NC)c2ccc(*)cc2OC1=C1)C1=O Chemical compound CC(CC1=C(c(c(*)c2)ccc2NC)c2ccc(*)cc2OC1=C1)C1=O 0.000 description 1
- XLDRMKUGYBFFDY-OYSZZVJOSA-N C[C@](CC[C@H](C1)O)([C@@H]1C1)[C@@H](CC[C@H]2C3CC[C@@H]2CCCC(NC(CCCCN)C(O)=O)=O)[C@H]3[C@@H]1O Chemical compound C[C@](CC[C@H](C1)O)([C@@H]1C1)[C@@H](CC[C@H]2C3CC[C@@H]2CCCC(NC(CCCCN)C(O)=O)=O)[C@H]3[C@@H]1O XLDRMKUGYBFFDY-OYSZZVJOSA-N 0.000 description 1
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical class [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 description 1
- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 1
- 108010007979 Glycocholic Acid Proteins 0.000 description 1
- DGABKXLVXPYZII-UHFFFAOYSA-N Hyodeoxycholic acid Natural products C1C(O)C2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 DGABKXLVXPYZII-UHFFFAOYSA-N 0.000 description 1
- SMEROWZSTRWXGI-UHFFFAOYSA-N Lithocholsaeure Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 SMEROWZSTRWXGI-UHFFFAOYSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- RFDAIACWWDREDC-UHFFFAOYSA-N Na salt-Glycocholic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(=O)NCC(O)=O)C)C1(C)C(O)C2 RFDAIACWWDREDC-UHFFFAOYSA-N 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- BHQCQFFYRZLCQQ-PGHAKIONSA-N allocholic acid Chemical compound C([C@@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-PGHAKIONSA-N 0.000 description 1
- DKPMWHFRUGMUKF-GDYCBZMLSA-N alpha-muricholic acid Chemical compound C([C@H]1[C@H](O)[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 DKPMWHFRUGMUKF-GDYCBZMLSA-N 0.000 description 1
- DKPMWHFRUGMUKF-CRKPLTDNSA-N beta-muricholic acid Chemical compound C([C@H]1[C@H](O)[C@@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 DKPMWHFRUGMUKF-CRKPLTDNSA-N 0.000 description 1
- 210000000013 bile duct Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- RUDATBOHQWOJDD-BSWAIDMHSA-N chenodeoxycholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 RUDATBOHQWOJDD-BSWAIDMHSA-N 0.000 description 1
- 229960001091 chenodeoxycholic acid Drugs 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000001143 conditioned effect Effects 0.000 description 1
- KXGVEGMKQFWNSR-LLQZFEROSA-N deoxycholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 KXGVEGMKQFWNSR-LLQZFEROSA-N 0.000 description 1
- 229960003964 deoxycholic acid Drugs 0.000 description 1
- NJDNXYGOVLYJHP-UHFFFAOYSA-L disodium;2-(3-oxido-6-oxoxanthen-9-yl)benzoate Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=CC=C1C1=C2C=CC(=O)C=C2OC2=CC([O-])=CC=C21 NJDNXYGOVLYJHP-UHFFFAOYSA-L 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- RFDAIACWWDREDC-FRVQLJSFSA-N glycocholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 RFDAIACWWDREDC-FRVQLJSFSA-N 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- DGABKXLVXPYZII-SIBKNCMHSA-N hyodeoxycholic acid Chemical compound C([C@H]1[C@@H](O)C2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 DGABKXLVXPYZII-SIBKNCMHSA-N 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- SMEROWZSTRWXGI-HVATVPOCSA-N lithocholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 SMEROWZSTRWXGI-HVATVPOCSA-N 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- NIQQIJXGUZVEBB-UHFFFAOYSA-N methanol;propan-2-one Chemical compound OC.CC(C)=O NIQQIJXGUZVEBB-UHFFFAOYSA-N 0.000 description 1
- DKPMWHFRUGMUKF-NTPBNISXSA-N omega-muricholic acid Chemical compound C([C@H]1[C@@H](O)[C@@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 DKPMWHFRUGMUKF-NTPBNISXSA-N 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000011877 solvent mixture Substances 0.000 description 1
- 150000003431 steroids Chemical group 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 238000001665 trituration Methods 0.000 description 1
- 238000002211 ultraviolet spectrum Methods 0.000 description 1
- RUDATBOHQWOJDD-UZVSRGJWSA-N ursodeoxycholic acid Chemical compound C([C@H]1C[C@@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 RUDATBOHQWOJDD-UZVSRGJWSA-N 0.000 description 1
- 229960001661 ursodiol Drugs 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J41/00—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
Definitions
- This invention relates to the production of bile acid derivatives and is particularly concerned with the production of bile acid derivatives containing a fluorescein moiety.
- Bile acid derivatives containing a fluorescein moiety are absorbed by the liver after injection into the blood and have a variety of uses.
- WO 99/07325 discloses the use of a cholyl lysyl fluorescein (CLF) derivative for determining liver function in a patient
- WO97/06829 disclose the use of a CLF derivative for visualising the bile duct and biliary leakage during an operation as an aid to the surgeon.
- the cholyl-lysine formate is then treated with NaOH, incubated and acidified with HCI, before the resultant solution is passed through a Sep-Pak C18 column, eluted with methanol and then evaporated to dryness to form the free cholyl-lysine.
- the cholyl-lysine is then dissolved in 0.1 M methanolic NaOH followed by addition of ether and trituration with ethyl acetate to form a sodium salt.
- the sodium salt is then dissolved in bicarbonate buffer and FITC (fluorescein isothiocyanate) in bicarbonate buffer followed by stirring for 18 hours at room temperature in the dark.
- Unreacted free FITC is removed by percolating the reaction mixture through Sep-Pak-C18 cartridges, the desired product is recovered by elution with methanol and further purification by TLC (thin layer chromatography) on silica gel plates using a solvent mixture of 3:1 v/v chloroform/methanol.
- TLC thin layer chromatography
- A is ⁇ -OH or ⁇ -OH
- B is ⁇ -H or ⁇ -H
- C is -H, ⁇ -OH or ⁇ -OH
- B and C together form a double bond
- D is -H, ⁇ -OH or ⁇ -OH
- E is -H, ⁇ - OH or ⁇ -OH
- Q is S or O
- X is an alkali metal (preferably Na or K) or ammonium ion
- n is 0 or 1
- m is 1 to 8, preferably 3 to 5 and most preferably 4; said method comprising the step of reacting (a) a bile acid derivative of the general formula (II):-
- A, B, C, D, E, Q, n and m are as defined above and each of X' and X" is -H, alkali metal or ammonium ion, provided that at least one of X' and X" is an alkali metal or an ammonium ion; with (b) an alkali of the formula X-OH, where X is as defined above, so as to produce said salt of the formula (I), and purifying the resultant mixture using reverse phase chromatography so as to isolate said compound of the formula (I).
- the reaction between compound (II) and the alkali of formula X-OH is effected in an aqueous-based reaction system to enable accurate control of pH. It is preferred for the pH of the aqueous-based reaction system to be at least 8 and more preferably in the range of 8 - 8.2.
- the bile acid derivative of the general formula (II) is produced by reacting a compound of the general formula (III)
- the compound of the general formula (III) may be produced by mixing a compound of the general formula (V):-
- A, B, C, D, E and n are as defined above with a non-polar solvent (such as acetone) and a tertiary amine (such as triethylamine), mixing ethyl chloroformate with the resultant mixture, and then adding an aqueous alkaline solution containing a compound of the general formula (VI):- H
- a non-polar solvent such as acetone
- a tertiary amine such as triethylamine
- BL is a blocking group (such as N-e-CBZ), followed by acidification, extraction and removal of the blocking group BL.
- a blocking group such as N-e-CBZ
- the purity of said salt is preferably at least 96%.
- the steroid moiety in the compound of the formula (I) may be based on cholic acid, chenodeoxycholic acid, deoxycholic acid, hyodeoxycholic acid, hyocholic acid, ⁇ -, ⁇ -, or ⁇ -muricholic acid, a nor-bile acid, lithocholic acid, 3 ⁇ -hydroxycholenoic acid, ursodeoxycholic acid, allocholic acid (5 ⁇ -cholan-24-oic acid), or the like
- the salt has the formula (VII):-
- a 10 litre stirred reaction vessel equipped with a lid, mechanical stirrer, anchor stirrer rod, dropping funnel, condenser, thermometer and funnel is charged sequentially with 858g acetone, 146g cholic acid and 50g triethylamine at ambient temperature with stirring. Stirring is continued for 30 minutes and 39g ethyl chloroformate is added dropwise over 20 minutes to the vessel while maintaining an internal temperature of 15- 25°C using ice/water bath cooling as necessary. Stirring is continued for 90 minutes. Following this the reaction vessel is charged with an aqueous solution containing 100g N-e-CBZ-L-lysine and 15g sodium hydroxide in 800g water. The resultant mixture is stirred vigorously for a further three hours, following which 550g 1M hydrochloric acid are added so as to attain a pH of 2.0-2.5.
- the contents of the reaction vessel are then transferred to a separating funnel and extracted twice with 900g ethyl acetate, combining the organic layers after each extraction.
- the combined organic extracts are then vacuum filtered.
- the filtered solution is then mixed with 5000g water and acidified with 1 M HCI to a pH of 2.0-2.5 before separation of the organic layer from the aqueous layer.
- the upper organic layer is then transferred to a container and charged with 150g magnesium sulphate, followed by filtration.
- the filtered organic layer is then dried in an evaporating flask on a rotary evaporator at a temperature of 45-50°C and then allowed to cool for twenty minutes.
- the yield of the Compound (VIII) is 200-21 Og, having a purity of > 90% (HPLC area%), typically 91-93%.
- a two litre reaction vessel equipped with mechanical stirrer, thermometer, dropping funnel and water-cooled condenser is charged with 1520g methanol at ambient temperature, and 190g N-e-CBZ-cholyl-L-lysine and 19g of 10% Pd/C (on a dry weight basis) at 15-20°C are added with stirring. Then, 342g formic acid is added, and the resultant reaction mixture is stirred for 24 hours at an ambient temperature. The reaction mixture is then vacuum filtered through 285g Celite into a flask. The reaction vessel is then washed with 475g methanol and the resultant washing combined with the filtered reaction mixture. The combined mixture is then poured into 13.3kg water to give a solution having a pH of 2.0-2.5.
- the resultant solution is then extracted twice with 3420g ethyl acetate in a separating funnel following which the resultant aqueous solution is adjusted to pH 4.5-5.0 by addition of 1283g 2.5M sodium hydroxide solution, followed by cooling of the solution to 0-4°C for four hours and then removing the resultant precipitate by vacuum filtration.
- the filtered precipitate is then dried in a vacuum oven at 45-50°C for four hours to afford cholyl-L-lysine (IX) as a white solid.
- the yield of cholyl-L- lysine (IX) is about 1 10-120g having a purity > 90%> (HPLC area %), typically 92-98%.
- a 2 litre glass reaction vessel equipped with a mechanical stirrer and a thermometer is charged with 750 ml 0.5 M sodium methoxide in methanol. After commencement of stirring, 10Og cholyl-L-lysine is added to the reaction vessel and the contents are stirred for 15 minutes. Then, 72g of fluorescein isothiocyanate isomer of 90 + % purity is added at ambient temperature. After stirring for one hour, a sufficient quantity of a 0.5M solution of sodium methoxide in methanol is added to ensure complete solution of the reaction mixture. Then, the contents of the reaction vessel are stirred at ambient temperature in the absence of light for 24 hours.
- a 20 litre glass flask equipped with a mechanical stirrer and a thermometer is charged with 9000g ethyl acetate.
- the contents of the first-mentioned reaction vessel are then transferred to the 20 litre vessel dropwise with stirring.
- the reaction mixture is stirred for a further 30 minutes and the resultant orange precipitate is vacuum filtered off.
- the resulting solid is then mixed with 500g methanol in a 2 litre reaction vessel with stirring, and stirring is continued until all the solid residue has dissolved.
- the resultant solution is transferred dropwise to a 20 litre vessel charged with 4500g acetone with stirring.
- the mixture is then stirred for a further 30 minutes, after which the precipitate is vacuum filtered and the methanol-acetone treatment is repeated once more followed by vacuum filtration.
- the resultant precipitate is dried under vacuum 45-50°C to constant weight to product compound (X).
- the yield of cholyl-L-lysine fluorescein di-sodium salt (X) is about 150- 170g having a purity > 80 HPLC area %, typically 83-89 HPLC area %.
- a column having a 4 inch (100 mm) bore is charged with 1.80kg Envi 18 reverse phase silica and pre-conditioned with 9 litres of 5% v/v solution of acetonitrile in water using nitrogen pressure and a bottom run-off valve as required to maintain a flow of approximately 200ml/min through the silica bed. This mobile phase is run through the bed until the top of the bed is just wetted.
- the collected fractions can then be combined according to purity so that one set of combined fractions contains compound (VII) at a purity of at least 96% with no single impurity being present in an amount greater than 0.5%, another set of combined fractions contains compound (VII) at a purity of at least 96% with at least one impurity being present in an amount greater than 0.5%, and further fractions of lesser purity.
- the pooled fractions are then freeze-dried or rotary evaporated to afford homogeneous compound (VII).
- the chromatography yields about 18g (40g loading) of cholyl-L-lysine fluorescein tri-sodium salt (compound VII) having a purity > 96 HPLC area %, typically 98-99.5 HPLC area %.
- the UV spectrum, the FT-IR spectrum, the DSC data and the NMR data for compound (VII) are respectively shown in accompanying Figs. 1 to 4.
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Abstract
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU2001276541A AU2001276541A1 (en) | 2000-08-10 | 2001-08-08 | Method for the production of fluorescein bile acid derivatives |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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GB0019593A GB0019593D0 (en) | 2000-08-10 | 2000-08-10 | Production of bile acid derivatives |
GB0019593.3 | 2000-08-10 |
Publications (1)
Publication Number | Publication Date |
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WO2002012267A1 true WO2002012267A1 (fr) | 2002-02-14 |
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Application Number | Title | Priority Date | Filing Date |
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PCT/GB2001/003559 WO2002012267A1 (fr) | 2000-08-10 | 2001-08-08 | Procede de production de derives d'acide biliaire a fluoresceine |
Country Status (3)
Country | Link |
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AU (1) | AU2001276541A1 (fr) |
GB (1) | GB0019593D0 (fr) |
WO (1) | WO2002012267A1 (fr) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB2467902A (en) * | 2009-02-12 | 2010-08-18 | Norgine Bv | Preparation of Cholyl-L-lysine from N-E-CBZ-cholyl-L-lysine |
EP2773653A4 (fr) * | 2011-11-02 | 2015-08-05 | Broad Inst Inc | Substrats fluorescents destinés à la détermination de l'activité enzymatique modifiée par la lysine |
US9745613B2 (en) | 2014-07-22 | 2017-08-29 | The Broad Institute, Inc. | Compounds, substrates and methods related to histone deacetylases |
WO2018221479A1 (fr) * | 2017-05-29 | 2018-12-06 | 五稜化薬株式会社 | Nouvelle substance fluorescente pour analyse de fonction de transport |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1997006829A1 (fr) * | 1995-08-19 | 1997-02-27 | The University Of Birmingham | Utilisation de derives de l'acide biliaire |
US5952187A (en) * | 1995-12-01 | 1999-09-14 | Oxis International, Inc. | Topiramate immunoassay |
US6013802A (en) * | 1995-02-06 | 2000-01-11 | Molecular Probes, Inc. | Fluorescent conjugates of metal-chelating nitrogen heterocycles |
-
2000
- 2000-08-10 GB GB0019593A patent/GB0019593D0/en not_active Ceased
-
2001
- 2001-08-08 WO PCT/GB2001/003559 patent/WO2002012267A1/fr active Application Filing
- 2001-08-08 AU AU2001276541A patent/AU2001276541A1/en not_active Abandoned
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6013802A (en) * | 1995-02-06 | 2000-01-11 | Molecular Probes, Inc. | Fluorescent conjugates of metal-chelating nitrogen heterocycles |
WO1997006829A1 (fr) * | 1995-08-19 | 1997-02-27 | The University Of Birmingham | Utilisation de derives de l'acide biliaire |
US5952187A (en) * | 1995-12-01 | 1999-09-14 | Oxis International, Inc. | Topiramate immunoassay |
Non-Patent Citations (3)
Title |
---|
M. K. HERRLEIN ET AL: "57. 3 -Amino-Modified Nucleotides Useful as Potent Chain Terminators for Current DNA Sequencing Methods", HELVETICA CHIMICA ACTA, vol. 77, no. 2, 23 March 1994 (1994-03-23), BASEL CH, pages 586 - 596, XP002183276 * |
MILKIEWICZ, P. ET AL: "Canalicular secretion of lysyl-fluorescein conjugated bile acids in 17.beta.-estradiol glucuronide (17.beta.EG) induced cholestasis: protective effect of s-adenosyl-L-methionine (SAMe)", BILE ACIDS HEPATOBILIARY DIS., PROC. FALK WORKSHOP (2000), MEETING DATE 1999, 72-84. EDITOR(S): NORTHFIELD, TIM C. PUBLISHER: KLUWER ACADEMIC PUBLISHERS, DORDRECHT, NETH., XP001039804 * |
MILLS C O ET AL: "CHOLYL-LYSYLFLUORESCEIN: SYNSTHESIS, BILIARY EXCRETION IN VIVO AND DURING SINGLE-PASS PERFUSION OF ISOLATED PERFUSED RAT LIVER", BBA - GENERAL SUBJECTS, ELSEVIER SCIENCE PUBLISHERS, NL, vol. 1115, no. 2, 1991, pages 151 - 156, XP000615215, ISSN: 0304-4165 * |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB2467902A (en) * | 2009-02-12 | 2010-08-18 | Norgine Bv | Preparation of Cholyl-L-lysine from N-E-CBZ-cholyl-L-lysine |
WO2010092381A1 (fr) | 2009-02-12 | 2010-08-19 | Norgine Bv | Procédé de préparation de cholyl-l-lysine |
CN102317304A (zh) * | 2009-02-12 | 2012-01-11 | 诺金公司 | 胆酰-l-赖氨酸的制备方法 |
JP2012517463A (ja) * | 2009-02-12 | 2012-08-02 | ノージーン ビーブイ | コリル−l−リシンの製造方法 |
EP2773653A4 (fr) * | 2011-11-02 | 2015-08-05 | Broad Inst Inc | Substrats fluorescents destinés à la détermination de l'activité enzymatique modifiée par la lysine |
US9856509B2 (en) | 2011-11-02 | 2018-01-02 | The Broad Institute, Inc. | Fluorescent substrates for determining lysine modifying enzyme activity |
US9745613B2 (en) | 2014-07-22 | 2017-08-29 | The Broad Institute, Inc. | Compounds, substrates and methods related to histone deacetylases |
WO2018221479A1 (fr) * | 2017-05-29 | 2018-12-06 | 五稜化薬株式会社 | Nouvelle substance fluorescente pour analyse de fonction de transport |
Also Published As
Publication number | Publication date |
---|---|
GB0019593D0 (en) | 2000-09-27 |
AU2001276541A1 (en) | 2002-02-18 |
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