WO2015042759A1 - Carboxymethyl-hydroxypropyl-β-cyclodextrin and preparation method thereof - Google Patents
Carboxymethyl-hydroxypropyl-β-cyclodextrin and preparation method thereof Download PDFInfo
- Publication number
- WO2015042759A1 WO2015042759A1 PCT/CN2013/001333 CN2013001333W WO2015042759A1 WO 2015042759 A1 WO2015042759 A1 WO 2015042759A1 CN 2013001333 W CN2013001333 W CN 2013001333W WO 2015042759 A1 WO2015042759 A1 WO 2015042759A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cyclodextrin
- hydroxypropyl
- carboxymethyl
- substitution
- product
- Prior art date
Links
- 238000002360 preparation method Methods 0.000 title abstract description 23
- 229920000858 Cyclodextrin Polymers 0.000 claims abstract description 98
- 238000006467 substitution reaction Methods 0.000 claims abstract description 77
- -1 hydroxypropyl Chemical group 0.000 claims abstract description 62
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical class OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 claims abstract description 50
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 claims abstract description 40
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 claims abstract description 38
- 239000001116 FEMA 4028 Substances 0.000 claims abstract description 31
- 229960004853 betadex Drugs 0.000 claims abstract description 31
- 239000000546 pharmaceutical excipient Substances 0.000 claims abstract description 6
- 239000003814 drug Substances 0.000 claims description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 28
- VEUUMBGHMNQHGO-UHFFFAOYSA-N ethyl chloroacetate Chemical compound CCOC(=O)CCl VEUUMBGHMNQHGO-UHFFFAOYSA-N 0.000 claims description 26
- 235000011175 beta-cyclodextrine Nutrition 0.000 claims description 25
- 238000000034 method Methods 0.000 claims description 18
- 238000003756 stirring Methods 0.000 claims description 15
- 239000000203 mixture Substances 0.000 claims description 14
- GOOHAUXETOMSMM-UHFFFAOYSA-N Propylene oxide Chemical compound CC1CO1 GOOHAUXETOMSMM-UHFFFAOYSA-N 0.000 claims description 11
- 238000006243 chemical reaction Methods 0.000 claims description 11
- 239000002585 base Substances 0.000 claims description 7
- 239000003513 alkali Substances 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
- 239000004375 Dextrin Substances 0.000 claims description 2
- 229920001353 Dextrin Polymers 0.000 claims description 2
- 239000007806 chemical reaction intermediate Substances 0.000 claims description 2
- 235000019425 dextrin Nutrition 0.000 claims description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 2
- 229940124531 pharmaceutical excipient Drugs 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 239000008194 pharmaceutical composition Substances 0.000 claims 2
- 230000002949 hemolytic effect Effects 0.000 abstract description 15
- 231100000053 low toxicity Toxicity 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 121
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 59
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 49
- 206010018910 Haemolysis Diseases 0.000 description 36
- 230000008588 hemolysis Effects 0.000 description 36
- 229940079593 drug Drugs 0.000 description 33
- 229940125396 insulin Drugs 0.000 description 25
- 102000004877 Insulin Human genes 0.000 description 24
- 108090001061 Insulin Proteins 0.000 description 24
- 238000012360 testing method Methods 0.000 description 23
- 229930101531 artemisinin Natural products 0.000 description 21
- BLUAFEHZUWYNDE-NNWCWBAJSA-N artemisinin Chemical compound C([C@](OO1)(C)O2)C[C@H]3[C@H](C)CC[C@@H]4[C@@]31[C@@H]2OC(=O)[C@@H]4C BLUAFEHZUWYNDE-NNWCWBAJSA-N 0.000 description 21
- 229960004191 artemisinin Drugs 0.000 description 21
- 239000000243 solution Substances 0.000 description 17
- 150000001875 compounds Chemical class 0.000 description 16
- 238000005481 NMR spectroscopy Methods 0.000 description 15
- RXUWDKBZZLIASQ-UHFFFAOYSA-N Puerarin Natural products OCC1OC(Oc2c(O)cc(O)c3C(=O)C(=COc23)c4ccc(O)cc4)C(O)C(O)C1O RXUWDKBZZLIASQ-UHFFFAOYSA-N 0.000 description 15
- 125000001424 substituent group Chemical group 0.000 description 15
- 229960002963 ganciclovir Drugs 0.000 description 14
- HKEAFJYKMMKDOR-VPRICQMDSA-N puerarin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1C1=C(O)C=CC(C2=O)=C1OC=C2C1=CC=C(O)C=C1 HKEAFJYKMMKDOR-VPRICQMDSA-N 0.000 description 14
- IRSCQMHQWWYFCW-UHFFFAOYSA-N ganciclovir Chemical compound O=C1NC(N)=NC2=C1N=CN2COC(CO)CO IRSCQMHQWWYFCW-UHFFFAOYSA-N 0.000 description 13
- 238000002347 injection Methods 0.000 description 13
- 239000007924 injection Substances 0.000 description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 12
- 230000000694 effects Effects 0.000 description 12
- 229940090044 injection Drugs 0.000 description 12
- 238000001228 spectrum Methods 0.000 description 12
- KWYUFKZDYYNOTN-UHFFFAOYSA-M potassium hydroxide Substances [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 11
- 239000000523 sample Substances 0.000 description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 238000004128 high performance liquid chromatography Methods 0.000 description 9
- 230000002829 reductive effect Effects 0.000 description 9
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 8
- 239000006069 physical mixture Substances 0.000 description 7
- DNIAPMSPPWPWGF-GSVOUGTGSA-N (R)-(-)-Propylene glycol Chemical compound C[C@@H](O)CO DNIAPMSPPWPWGF-GSVOUGTGSA-N 0.000 description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 239000012535 impurity Substances 0.000 description 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N monopropylene glycol Natural products CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 230000003287 optical effect Effects 0.000 description 6
- 235000013772 propylene glycol Nutrition 0.000 description 6
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 5
- 229940097362 cyclodextrins Drugs 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 239000012467 final product Substances 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- ODLHGICHYURWBS-LKONHMLTSA-N trappsol cyclo Chemical compound CC(O)COC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)COCC(O)C)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1COCC(C)O ODLHGICHYURWBS-LKONHMLTSA-N 0.000 description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 239000012981 Hank's balanced salt solution Substances 0.000 description 4
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 238000000502 dialysis Methods 0.000 description 4
- 229930003935 flavonoid Natural products 0.000 description 4
- 235000017173 flavonoids Nutrition 0.000 description 4
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 238000011068 loading method Methods 0.000 description 4
- 238000002844 melting Methods 0.000 description 4
- 230000008018 melting Effects 0.000 description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- 230000035515 penetration Effects 0.000 description 4
- 108090000765 processed proteins & peptides Proteins 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 230000032258 transport Effects 0.000 description 4
- CUJVBAPGYBSBHJ-YWBSARSQSA-N 2-[[(1R,3R,5R,6S,8R,10R,11S,13R,15R,16S,18R,20R,21R,23R,25R,26R,28R,30R,31R,33R,35R,36R,37R,38R,39R,40R,41R,42R,43R,44R,45R,46R,47R,48R,49R)-36,38,40,42-tetrakis(carboxymethoxy)-10,15-bis(carboxymethoxymethyl)-37,39,41,43,44,45,46,47,48,49-decahydroxy-20,25,30,35-tetrakis(hydroxymethyl)-2,4,7,9,12,14,17,19,22,24,27,29,32,34-tetradecaoxaoctacyclo[31.2.2.23,6.28,11.213,16.218,21.223,26.228,31]nonatetracontan-5-yl]methoxy]acetic acid Chemical compound OC[C@H]1O[C@@H]2O[C@H]3[C@H](O)[C@@H](O)[C@H](O[C@@H]3COCC(O)=O)O[C@H]3[C@H](O)[C@@H](O)[C@H](O[C@@H]3COCC(O)=O)O[C@H]3[C@H](O)[C@@H](O)[C@H](O[C@@H]3COCC(O)=O)O[C@@H]3[C@@H](CO)O[C@H](O[C@@H]4[C@@H](CO)O[C@H](O[C@@H]5[C@@H](CO)O[C@H](O[C@H]1[C@H](OCC(O)=O)[C@H]2O)[C@H](O)[C@H]5OCC(O)=O)[C@H](O)[C@H]4OCC(O)=O)[C@H](O)[C@H]3OCC(O)=O CUJVBAPGYBSBHJ-YWBSARSQSA-N 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 238000006073 displacement reaction Methods 0.000 description 3
- 230000002708 enhancing effect Effects 0.000 description 3
- 150000002215 flavonoids Chemical class 0.000 description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000010992 reflux Methods 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- DOMXUEMWDBAQBQ-WEVVVXLNSA-N terbinafine Chemical compound C1=CC=C2C(CN(C\C=C\C#CC(C)(C)C)C)=CC=CC2=C1 DOMXUEMWDBAQBQ-WEVVVXLNSA-N 0.000 description 3
- 238000002076 thermal analysis method Methods 0.000 description 3
- YCHXNMPJFFEIJG-UHFFFAOYSA-N 3-methyloxiran-2-one Chemical compound CC1OC1=O YCHXNMPJFFEIJG-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 101000986989 Naja kaouthia Acidic phospholipase A2 CM-II Proteins 0.000 description 2
- XPUJQEGMNQWMJI-UHFFFAOYSA-N OCC(=O)O.[Na] Chemical compound OCC(=O)O.[Na] XPUJQEGMNQWMJI-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Natural products CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 2
- 230000007059 acute toxicity Effects 0.000 description 2
- 231100000403 acute toxicity Toxicity 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000005100 correlation spectroscopy Methods 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 229940093181 glucose injection Drugs 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 231100000956 nontoxicity Toxicity 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 238000007911 parenteral administration Methods 0.000 description 2
- 229920001606 poly(lactic acid-co-glycolic acid) Polymers 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 238000001896 rotating frame Overhauser effect spectroscopy Methods 0.000 description 2
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 description 2
- 235000010234 sodium benzoate Nutrition 0.000 description 2
- 239000004299 sodium benzoate Substances 0.000 description 2
- 239000012265 solid product Substances 0.000 description 2
- 238000005063 solubilization Methods 0.000 description 2
- 230000007928 solubilization Effects 0.000 description 2
- 229960000699 terbinafine hydrochloride Drugs 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- YZOUYRAONFXZSI-SBHWVFSVSA-N (1S,3R,5R,6R,8R,10R,11R,13R,15R,16R,18R,20R,21R,23R,25R,26R,28R,30R,31S,33R,35R,36R,37S,38R,39S,40R,41S,42R,43S,44R,45S,46R,47S,48R,49S)-5,10,15,20,25,30,35-heptakis(hydroxymethyl)-37,39,40,41,42,43,44,45,46,47,48,49-dodecamethoxy-2,4,7,9,12,14,17,19,22,24,27,29,32,34-tetradecaoxaoctacyclo[31.2.2.23,6.28,11.213,16.218,21.223,26.228,31]nonatetracontane-36,38-diol Chemical compound O([C@@H]([C@H]([C@@H]1OC)OC)O[C@H]2[C@@H](O)[C@@H]([C@@H](O[C@@H]3[C@@H](CO)O[C@@H]([C@H]([C@@H]3O)OC)O[C@@H]3[C@@H](CO)O[C@@H]([C@H]([C@@H]3OC)OC)O[C@@H]3[C@@H](CO)O[C@@H]([C@H]([C@@H]3OC)OC)O[C@@H]3[C@@H](CO)O[C@@H]([C@H]([C@@H]3OC)OC)O3)O[C@@H]2CO)OC)[C@H](CO)[C@H]1O[C@@H]1[C@@H](OC)[C@H](OC)[C@H]3[C@@H](CO)O1 YZOUYRAONFXZSI-SBHWVFSVSA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 238000012404 In vitro experiment Methods 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 206010029155 Nephropathy toxic Diseases 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 102000008579 Transposases Human genes 0.000 description 1
- 108010020764 Transposases Proteins 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 231100000215 acute (single dose) toxicity testing Toxicity 0.000 description 1
- 238000011047 acute toxicity test Methods 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 239000005456 alcohol based solvent Substances 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000012752 auxiliary agent Substances 0.000 description 1
- 230000037429 base substitution Effects 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000001142 circular dichroism spectrum Methods 0.000 description 1
- 229940121657 clinical drug Drugs 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000009792 diffusion process Methods 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
- 238000007922 dissolution test Methods 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- 238000005570 heteronuclear single quantum coherence Methods 0.000 description 1
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 238000009830 intercalation Methods 0.000 description 1
- 230000002687 intercalation Effects 0.000 description 1
- 230000000622 irritating effect Effects 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 231100000682 maximum tolerated dose Toxicity 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- JWHHANVGNNWIRI-UHFFFAOYSA-N methanol phosphoric acid hydrate Chemical compound O.OC.OP(O)(O)=O JWHHANVGNNWIRI-UHFFFAOYSA-N 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000007694 nephrotoxicity Effects 0.000 description 1
- 231100000417 nephrotoxicity Toxicity 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 125000001436 propyl group Chemical class [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 238000007363 ring formation reaction Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 229960003885 sodium benzoate Drugs 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 1
- 238000012353 t test Methods 0.000 description 1
- 229960002722 terbinafine Drugs 0.000 description 1
- 238000001757 thermogravimetry curve Methods 0.000 description 1
- 230000005945 translocation Effects 0.000 description 1
- 230000032895 transmembrane transport Effects 0.000 description 1
- 238000002211 ultraviolet spectrum Methods 0.000 description 1
- 231100000925 very toxic Toxicity 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61K47/40—Cyclodextrins; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
- C08B37/0012—Cyclodextrin [CD], e.g. cycle with 6 units (alpha), with 7 units (beta) and with 8 units (gamma), large-ring cyclodextrin or cycloamylose with 9 units or more; Derivatives thereof
Definitions
- the present invention relates to a carboxy-methyl and hydroxypropyl mixed-substituted oxime-cyclodextrin derivative carboxymethyl group for pharmaceutical excipients. - Hydroxypropyl cyclodextrin and a process for its preparation. Background technique
- ⁇ -cyclodextrin is a cyclic oligo natural product obtained by cyclization of cyclodextrin glucan transposase to obtain a cyclic oligosaccharide.
- the molecules are connected by a glycoside bond to form a frustoconical body having a hollow inner cavity, and 21 hydroxyl groups are distributed around the periphery of the pyramid cavity. Its hydrophobic and extracellular hydrophilic properties make it possible for 3-CD to form a supramolecular (compound) with weak organic molecules.
- Organic small molecules have improved physical and chemical properties due to the formation of supramolecules, and have high research and application value in agricultural, pharmaceutical, food and cosmetic applications.
- P-CD solubility of P-CD is small (1.85%), which affects the ability of inclusion and solubilization. It has obvious nephrotoxicity in parenteral administration and has strong hemolysis effect. It is only suitable for oral administration and cannot be used for parenteral administration. Therefore, it is important to modify the ⁇ -CD structure to create new derivatives, improve the solubility and enhance the inclusion ability, and reduce the hemolysis.
- the pharmaceutically acceptable cyclodextrin derivative must have good water solubility, low hemolytic activity, low toxicity, and inclusion of solubilizing ability. Studies have shown that both hydrophilic hydroxypropyl and sulfobutyl groups are good substituents for the modification of ⁇ -CD, and the derivatives thus prepared are hydroxypropyl- ⁇ -cyclodextrin ( ⁇ - ⁇ -CD) and sulfonate. Butyl- ⁇ -cyclodextrin (SBE--CD) has become a commonly used medicinal cyclodextrin and is widely used in the pharmaceutical and food industries. Carboxymethyl- ⁇ -cyclodextrin (CM-p-CD) is commercially available as a commercially available product, but there are currently no large industrial applications.
- M 1149 ⁇ nysg-moi 1 )
- the drug-loading efficiency is high, but the hemolysis effect of the aqueous solution is obvious and the irritating effect is strong
- the sulfobutyl-e-cyclodextrin is insoluble in the organic solvent, and the molecular weight is relatively high.
- the drug-loading efficiency is low
- substituents can significantly enhance the solubility and inclusion capacity of ⁇ -CD derivatives.
- the change in the type of substituents has little effect on the solubility properties, but the change in inclusion ability and hemolysis
- the impact is more complicated, and the inclusion ability is related to the type of substituent (including the structure of the material), and also closely related to the structure and functional group type of the encapsulated drug molecule; the change of hemolysis depends on the type of substituent.
- ⁇ - ⁇ -CD is less hemolytic than maternal ⁇ -CD, and has little change in hemolysis in the wider DS range (2-10).
- the hemolytic effect of SBE-P-CD decreased significantly and fluctuated greatly with the change of DS.
- the increase of DS was beneficial to reduce hemolysis.
- the carboxymethyl group can reduce the hemolysis of the product, and the relationship between DS and hemolysis is still unclear. Although the low concentration shows significant hemolysis, it has lower hemolytic properties than HP- ⁇ -CD at high concentrations.
- the molecular weight of pharmaceutically acceptable cyclodextrin derivatives is also an important factor that must be considered in practical applications.
- an excessively large molecular weight will make the amount of prescription excipients too high, and the effect of the actual function of the cyclodextrin product becomes an important reason for limiting its practicability.
- the design and preparation of lower molecular weight cyclodextrin derivatives is not only of great significance, but also one of the technical difficulties in the development of pharmaceutical cyclodextrins. Summary of the invention
- the technical problem to be solved by the present invention is to improve the water solubility of ⁇ -cyclodextrin and to prepare a ⁇ -cyclodextrin derivative which is safe, effective and has a relatively low molecular weight.
- the technical method is to introduce two smaller hydrophilic substituents in the ⁇ -cyclodextrin precursor to improve the performance of ⁇ -cyclodextrin, improve product safety, and enhance product practicability.
- the technical solution of the present invention is:
- the carboxymethyl-hydroxypropyl-e-cyclodextrin described in the present invention contains a plurality of compounds of the above molecular structure.
- CH 2 CH(OH)CH 3 , H" means that when R ⁇ OR is CH 2 COO-X, among the 7 R 3 groups in the above structural formula, part of the R 3 group is CH 2 CH(OH CH 3 , another part of the R 3 group is H. The rest of the similar expressions are all this.
- Carboxymethyl-hydroxypropyl- ⁇ -cyclodextrin can be represented by CMn-HPm-p-CD, the structure of which is carboxymethyl (-CH2COO-X) and 2-hydroxypropyl (-CH 2 CH(OH)CH 3 ) are ether bond-substituted derivatives formed by simultaneous attachment to a cyclodextrin precursor, ie, the cyclodextrin derivative has both a carboxyl group Base and hydroxypropyl two substituent groups.
- the average number of substituents per mole of ⁇ -cyclodextrin is the average degree of substitution (DS): ⁇ carboxymethyl and m hydroxypropyl.
- n is the average degree of substitution of carboxymethyl groups
- m + n - Z is the total average degree of substitution; the mixed substituted product must be nl and ml.
- the substitution position of the hydroxypropyl or carboxymethyl group of the product is randomly substituted. Derivative of cyclodextrin at position 2, or position 3 or position 6.
- Carboxymethyl and hydroxypropyl mixed substituted cyclodextrin derivatives abbreviation: carboxymethyl-hydroxypropyl-e-cyclodextrin ), not hydroxypropyl-cyclodextrin and carboxymethyl- a simple mixture of both ⁇ -cyclodextrin.
- the average degree of substitution n and tn represented by the structural characteristics of the cyclodextrin derivative also contain the actual average value within ⁇ 0.5, and the average degree of substitution for each of the optional derivatives is
- the carboxymethyl-hydroxypropyl-e-cyclodextrin designed by the present invention is ⁇ -cyclodextrin, ethyl chloroacetate It is prepared by using 1,2-epoxypropionam as a raw material, adding ⁇ -cyclodextrin and a catalytic amount of alkali in water, and adding ethyl chloroacetate and 1,2-epoxypropanoid dropwise after stirring at an appropriate temperature. After purification, a series of designs CMn-HPm-P-CDo can be obtained.
- the product with the desired degree of substitution can be obtained: with the increase of the reagent ratio, the degree of product substitution. That is to say, it is continuously improved, and the ratio of substitution of reagents is also reduced.
- the reagent charge can be obtained by adding 2.0 times the molar amount of ethyl chloroacetate (relative to cyclodextrin) and 1.5 times or more by mole of 1,2-epoxypropane.
- a mixed substitution product of m ⁇ l; a molar amount of 7 times cyclodextrin plus 2 times of ethyl chloroacetate reagent (molar) is the total amount of catalyst base.
- the present invention provides a preparation method of the above carboxymethyl-hydroxypropyl- ⁇ -cyclodextrin, which is prepared by a method of base-catalyzed continuous reaction, and the specific reaction step is one of the following two schemes: 1: Take ⁇ -cyclodextrin and add 2.2 times mass of water and 7.0 times molar amount of base to 0-cyclodextrin, stir to dissolve, and add 7.0 times molar amount of 1,2-epoxypropane to react at room temperature.
- the parent cyclodextrin is used as a raw material, and the reaction intermediate is mixed and replaced by a continuous feeding reaction step without separation.
- the present invention provides the pharmaceutical use of any of the above carboxymethyl-hydroxypropyl cyclodextrin, comprising the pharmaceutically acceptable form of carboxymethyl-hydroxypropyl- ⁇ -cyclodextrin
- the agent is applied to a composition comprising an active molecule preparation.
- the present invention provides the above oral preparation product containing the carboxymethyl-hydroxypropyl-3-cyclodextrin composition for therapeutic or health care use.
- the present invention provides a composition of the above carboxymethyl-hydroxypropyl cyclodextrin for use as a non-oral preparation product for therapeutic or health care purposes.
- the present invention provides the use of any of the above carboxymethyl-hydroxypropyl cyclodextrin as a pharmaceutical adjuvant.
- the hemolytic effect of the synthesized carboxymethyl-hydroxypropyl- ⁇ -cyclodextrin of the invention is significantly lower than that of the corresponding monosubstituted derivatives ⁇ - ⁇ -CD and CM-p-CD, with low hemolysis rate and substantially no toxicity. . No harmful residues, the product is safe. Not only the product itself has a small hemolysis effect, but also can significantly reduce the hemolysis rate of the encapsulated drug and improve the safety of administration.
- the product has excellent solubility, solubility in water is 50%, and organic solvent also shows good solubility. It has laid a material foundation for improving the solubility of poorly soluble drugs by inclusion technology, and has a broad prospect in the application of poorly soluble pharmaceutical preparations.
- the product of the invention has the appropriate ability to contain drugs, and has good inclusion properties for various types of drugs, including polypeptide macromolecules. It has important application value for improving drug solubility, enhancing absorption and improving bioavailability.
- the carboxymethyl-hydroxypropyl- ⁇ -cyclodextrin prepared by the invention has a moderate average molecular weight, which is equivalent to the molecular weight of the existing product ⁇ - ⁇ -CD, but the hemolytic effect is significantly lower than that of ⁇ - ⁇ -CD, thereby The application feasibility of the product of the invention is greatly improved.
- the preparation and analysis method of the invention is simple, the preparation yield is high, and the quality is stable.
- the product of the series was prepared in a weight yield of 100%. Further, the preparation method of the present invention is easy to obtain (commercially available p-CD, 1,2-propylene oxide, ethyl chloroacetate), and the production cost is low. In the preparation process, the feed ratio can be adjusted to obtain the Products that need to be replaced.
- the product of the invention has stable properties, no toxicity, no flammable and explosive characteristics, no environmental pollution, no deterioration, easy storage and easy transportation.
- Figure 6 Product differential thermogram: a: ⁇ -CD; b: ⁇ - ⁇ -CD; c: CM-P-CD; d: HP-p-CD/p-CD/CM-p-CD E; CM 2 -HP 6 - -CD
- Figure 7 HPLC chromatogram of product
- Figure 8 HPLC chromatogram of three physical mixtures of p-CD, CM-p-CD and ⁇ - ⁇ -CD
- Figure 9 Artemisinin , CM 3 -HP 6 -p-CD, artemisinin and CM 3 -HP 6 -p-CD physical mixture, CM 3 -HP 6 -P-CD / artemisinin inclusion complex four samples comparison DTA map: a : artemisinin; b: CM 3 -HP 6 -p-CD ; c: artemisinin / CM 3 -HP 6 -p_CD physical mixture; d: CM 3 -HP 6 -P-CD / artemisinin inclusion
- a three-neck round bottom flask was equipped with a constant pressure funnel, a reflux condenser, and a thermometer. 50 ml of purified water was added thereto, and stirring was started. Then, 0.02 ⁇ 1 ⁇ -cyclodextrin (22.7 g) and 0.3 mol of solid NaOH (12.0) were slowly added thereto with stirring. g), after the system is dissolved, slowly heat, keep 80 ° C, then slowly add 0.08 mol of ethyl chloroacetate (9.8 g), add about 4 h, continue to stir and reflux for 2 h and then cool to room temperature.
- DTA spectrum 95 'C dehydration endothermic peak, 270 ° C melting point, 300 ⁇ 335 ° C exothermic decomposition.
- Example 4 It was basically the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.22, 0.04, and 0.05 mol, respectively. The final product yield was 98.5%. The average degree of substitution of carboxymethyl group was 1.2 by 1 H NMR, and the average degree of substitution of hydroxypropyl group was 1.8. The product was abbreviated as CM!-HPz-p-CDa. Example 4
- Example 2 Basically the same as in Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.22, 0.04, and 0.08 mol, respectively.
- the yield of the final product was 99.1%.
- the average degree of substitution of the carboxymethyl group was 1.1 by the 1 H NMR, and the average degree of substitution of the hydroxypropyl group was 3.4.
- the product was abbreviated as CM!-HPHi-CDa.
- Example 2 The same as Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.26, 0.06, and 0.06 mol, respectively.
- the yield of the final product was 100.7%.
- the average degree of substitution of carboxymethyl group was 1.9 by 1 H NMR, and the average degree of substitution of hydroxypropyl group was 2.2.
- the product was abbreviated as CM 2 -HPH3-CD.
- Example 8 the product of potassium salt
- Example 2 It was basically the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.22, 0.04, and 0.12 mol, respectively.
- the yield of the product was 106.2%.
- the average degree of substitution of the carboxymethyl group was 1.3 by the 1 H NMR, and the average degree of substitution of the hydroxypropyl group was 4.6.
- the product was abbreviated as CMi-HPs-P-CDn.
- the yield was 110.9%, the average degree of substitution of carboxymethyl group was 2.9 by 1 H NMR, and the average degree of substitution of hydroxypropyl group was 2.8.
- the product was abbreviated as CM 3 -HP 3 -P-CD.
- Example 10 was substantially the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate and 1,2-propylene oxide were 0.26, 0.06, and 0.11 mol, respectively.
- Example 11 is basically the same as Example 1, except that NaOH is changed to KOH, and the amounts of KOH, ethyl chloroacetate, and 1,2-epoxypropionam are 0.62, 0.24, and 0.08 mol, respectively.
- the product (X K, hydrazine), yield 119.9%, the average degree of substitution of carboxymethyl groups was 5.7 by 1 H NMR, and the average degree of substitution of hydroxypropyl groups was 3.2.
- Example 12 was substantially the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-propylene oxide were 0.22, 0.04, and 0.03 mol, respectively.
- the yield of the product was 93.4%.
- the average degree of substitution of the carboxymethyl group was 1.4 by the 1 H NMR, and the average degree of substitution of the hydroxypropyl group was 1.3.
- the product was abbreviated as CMT-HP!-P-CDa.
- Example 13 was basically the same as Example 1. However, the amounts of NaOH, ethyl chloroacetate and 1,2-epoxypropanone were 0.5, 0.18 and 0.13 mol, respectively.
- Example 14 was basically the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate and 1,2-propylene oxide were 0.7, 0.24, and 0.03 mol, respectively.
- the final yield of the product was 113.8%.
- the average degree of substitution of the carboxymethyl group was 5.6 by 1 H NMR, and the average degree of substitution of the hydroxypropyl group was 1.3.
- the product was abbreviated as CM 6 -HP ⁇ -CD.
- Example 15 was substantially the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate and 1,2-propylene oxide were 0.26, 0.06, and 0.10 mol, respectively.
- the final yield of the product was 108.7%.
- the average degree of substitution of the carboxymethyl group was 1.8 by the 1 H NMR, and the average degree of substitution of the hydroxypropyl group was 3.9.
- the product was abbreviated as CM 2 -HP 4 -P-CD.
- Example 16 was substantially the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.45, 0.15, and 0.03 mol, respectively.
- the yield of the final product was 105.4%.
- the average substitution degree of carboxymethyl group was 3.8 by 1 H NMR, and the average degree of substitution of hydroxypropyl group was 1.3.
- the product was abbreviated as
- Example 17 Inclusion of a product with artemisinin and preparation of a clathrate.
- CM-p-CD ⁇ 0.0102 ⁇ + 9.5804 0.9957 939.2
- CM 3 -HP 6 - -CD ⁇ - 0.0069 ⁇ + 10.667 0.9934 1545.9
- the inclusion constant Ka determination results show that the inclusion ability of CM 3 -HP 6 -P-CD for artemisinin is significantly stronger than the existing product ⁇ - ⁇ - CD and CM-p-CD, CM-HP-p-CD binary substituted cyclodextrin derivatives facilitate drug inclusion, and the corresponding drug inclusion complexes are more stable and easier to manufacture.
- 10 g of CM 3 -HPH3-CD and 1.70 g of artemisinin were accurately weighed, mixed with an appropriate amount of pure water for 3 hours, and dried under reduced pressure at 50 ° C to obtain a white solid clathrate (molar ratio of 1:1). Take artemisinin, CM 3 -HP 6 -P-CD, artemisinin and CM 3 -HP 6 -P-CD physical mixture,
- CM 3 -HPH3-CD/artemisinin inclusion complex four samples each about 5.0mg, for differential scanning thermal analysis: ⁇ 1 2 ⁇ 3 reference, range ⁇ 50 ⁇ , temperature range 40 ° C ⁇ 400 ° C, heating rate At 10 ° C / min, the DTA map was obtained (Fig. 9).
- Solubility determination Artemisinin, artemisinin/HP-p-CD inclusion complex (prepared by the same method, molar ratio 1: 1.16), CM 3 -HP 6 -P-CD/artemisinin inclusion complex Molar ratio 1: 1 ) The solubility of the sample was measured at 25 ° C. Results: Artemisinin Ol lmg'ml- ⁇ CM 3 -HP 6 -P_CD inclusion complex l.OSmg'ml- ⁇ - ⁇ -CD inclusion complex 0.53 Mg'm, CM 3 -HP 6 -p-CD increased the solubility of artemisinin by 9.55 times, while ⁇ - ⁇ -CD only increased the solubility by 4.81 times.
- the iHNMR spectrum shows: due to the influence of the substituent, the proton of the cyclodextrin glucose ring in the product structure
- the displacements are interlaced to form a set of multiple peaks (similar to the ⁇ - ⁇ -CD spectrum), rather than as clear and easy to distinguish as ⁇ -CD, the -dH proton of the glucose ring is also superimposed by the doublet in ⁇ -CD.
- CM 2 -HP 3 -P-CD sample was weighed and mixed with the ganciclovir bulk drug 1.0 g, and added to 4 ml of water for 0.5 h, and then dried under reduced pressure at 45 Torr to obtain 1 g of a powdery product.
- Weigh the appropriate amount of ganciclovir inclusion compound dissolve in glucose injection, filter with 0.2 ⁇ filter, and prepare 100ml of inclusion compound injection of ganciclovir concentration 1.0mg_m.
- the invention adopts HP-p-CD, CM-p-CD and ⁇ -CD as control, and carries out in vitro experiments of CM-HP_p_CD on insulin incorporation, promotion of Caco-2 cell insulin in vitro, and cytotoxicity.
- CM-HP-p-CD has a stronger inclusion of insulin and promotes transmembrane transport of insulin, and is essentially non-toxic to membrane cells.
- CM-HP-P-CD has a good hypoglycemic ability to promote insulin.
- the control ⁇ - ⁇ -CD can be used not only as a preparation for chemical drugs, but also as a carrier for excellent protein macromolecular drugs, thus greatly expanding the application field of ⁇ - ⁇ -CD.
- Intensive strength Preparation and properties of carboxymethyl-hydroxypropyl cyclodextrin and peptide type drug-insulin inclusion compound.
- the CM 2 -HP 6 -p-CD of the present invention is measured by the commercially available HP-P-CD, CM_p_CD and ⁇ -CD as the control, and the apparent inclusion constant Ka of insulin is determined. The results are shown in the following table:
- the cells were seeded in 12-well Transwell (1.12 cm 2 , 0.4 ⁇ ), the plate density was 2 ⁇ 10 5 'mL- 1 , and the medium added to the apical and basal ends was 0.5 mL and 1.5 mL, respectively, and the cells were cultured for about 3 weeks. Fusion differentiation.
- HBSS was used to wash away the metabolites adhering to the cell surface, and the permeation amount of sodium fluorescein did not exceed 4. ⁇ g-hr l -cm- 2 , and the TEER value was about 500 Q'cm- 2 .
- the Apical side chamber (referred to as the A side) was added with 15 Il ⁇ mL- 1 insulin and clathrate solution, and 1.5 mL HBSS was added to the Basolateral side chamber (B side) below the membrane, and the temperature was 50 rmin- 1 at 37 °C. Incubate on an air shaker, sample 0.5 ml from the BL pool at 30, 60, 90, 120, 150 min, and immediately add an equal amount of HBSS; determine the insulin content in the sample by HPLC. Repeat 3 times for each experiment. The penetration of insulin (in IU) versus time (t) gives the insulin penetration curve.
- CM 2 - HP 6 - -CD > CM- -CD> HP- -CD>p-CD the order is exactly the same as their insulin inclusion constant.
- CM 2 -HIVP-CD prepared according to the invention has the strongest effect of enhancing insulin penetration and translocation.
- Example 21 Preparation of carboxymethyl-hydroxypropyl- ⁇ -cyclodextrin and flavonoid drug-puerarin and hemolysis test. Weigh accurately 10g of the CM 3 -HP 3 -P-CD of the present invention is uniformly mixed with 2.8g of puerarin (molar ratio of 1:1), melted at 60 ° C, heated at a temperature of 1 h, and then cooled at 25 ° C.
- the product of the present invention can easily prepare a clathrate compound for the polyphenolic flavonoid drug, and it is not necessary to add any solvent auxiliary agent in the preparation process, and is particularly suitable for the preparation of the sensitive drug clathrate.
- CM 2 -HP 5 -p-CD inclusion complex mass ratio 5: 1
- the inclusion complex 6g dissolved in NaCl injection filtered with 0.2 ⁇ filter, prepared into 500ml of injection of puerarin concentration Z.Omg.ml- 1
- hemolysis test (literature method: "Guidelines for Drug Research Technology (2005)", Chinese medicine Science and Technology Press, 2006, P116-132) No obvious hemolysis (hemolytic rate ⁇ 5%) was observed, and the common commercial puerarin injection (puerarin concentration S.Omg'ml- 1 ) with the same concentration was observed. Hemolysis (hemolytic rate 09%). Tests have shown that the use of the CM-HP-P-CD prepared by the present invention can effectively reduce the hemolysis of the drug, thereby enhancing the safety of the clinical drug.
- IR spectrum of KBr pellets of different substituted carboxymethyl-hydroxypropyl-P-cyclodextrin products prepared by the present invention is compared with ⁇ -CD, and the CM-HP-p-CD products are 2969cm- 1 and 1461cm.
- - 1 has significant absorption (antisymmetric stretching vibration of hydroxypropyl group and symmetric bending vibration absorption), and 1420 cm- 1 and 1617 cm- 1 peaks indicate the presence of a carboxymethyl substituent.
- the products of different degrees of substitution of carboxymethyl-hydroxypropyl cyclodextrin prepared by the invention are all white amorphous powdery solids, which are easy to absorb moisture, and the products still contain 4% ⁇ 11% of inclusion water after sufficient drying. All products are thermally decomposed without a fixed melting point, and the decomposition point is gradually increased in the range of 230X to 280 °C as the degree of substitution (increase in molecular weight) increases.
- the sample (e) features: 90 ° C or less is wider than p-CD, CM-p-CD and ⁇ - ⁇ -CD.
- Table 2 the partial product substitution degree prepared by the present invention obtained by iH NMR is shown in Table 2:
- the carboxymethyl-hydroxypropyl-3-cyclodextrin prepared by the invention has the similar solubility characteristics of ⁇ - ⁇ -CD, and the series products are highly soluble in water, dilute acid/dilute lye, and also very soluble in methanol. / Ethanol aqueous solution, soluble in pure methanol / pure ethanol, difficult to dissolve or insoluble in common organic solvents such as n-hexane, ethyl acetate, chloroform, acetone, etc., the solubility properties of the products with different degrees of substitution are basically no difference.
- the solubility of carboxymethyl-hydroxypropyl-e-cyclodextrin in pure ethanol can be more than 2.5 times that of hydroxypropyl-sulfobutylcyclodextrin. Good solubility characteristics will be very beneficial for high fat-soluble drug inclusion complexes. Preparation, research and development of new systems for improving the performance of poorly soluble drugs Agent products have important application value.
- the solubility test results using CM 3 -HP 3 -p-CD as an example are shown in the table below.
- the carboxymethyl-hydroxypropyl- ⁇ -cyclodextrin prepared by the present invention is a carboxymethyl and hydroxypropyl mixed-substituted cyclodextrin derivative instead of hydroxypropyl- ⁇ -cyclodextrin.
- the CD has significantly different HPLC chromatographic characteristics, making it easy to identify the products of the invention.
- Chromatographic conditions column Amino column (4.6mm x 250mm, 5 m); Refractive detector; Mobile phase: Acetonitrile: Water (67: 33); Flow rate: l.Oml/min; Column temperature 30 °C. Accurately weigh P_CD, CM--CD.
- the product of the invention is a product of mixed substitution of hydroxypropyl and carboxymethyl, instead of hydroxypropyl cyclodextrin and carboxymethyl - A simple physical mixture of ⁇ -cyclodextrin.
- the carboxymethyl-hydroxypropyl-3-cyclodextrin of the invention is obtained by reacting ⁇ -cyclodextrin with ethyl chloroacetate and 1,2-epoxypropanone in a strong base in an aqueous solution, and the system impurities are residual Base-catalyzed hydrolysates of ⁇ -cyclodextrin and reagents: glycolic acid and 1,2-propanediol, using dialysis purification technology to adjust ⁇ , can greatly reduce the residual amount of ⁇ -cyclodextrin and 1,2-propanediol
- the glycolic acid has a small molecular weight and exists in the form of an anion to be more easily removed, so that the carboxymethyl-hydroxypropyl cyclodextrin product of the invention has high purity and good product dissolution and inclusion properties, and is significantly low.
- the main impurities and limits to be controlled in the carboxymethyl-hydroxypropyl- ⁇ -cyclodextrin product of the present invention are as follows: Main Impurities in Carboxymethyl-Hydroxypropyl Cyclodextrin Impurity Source Impurity Limit Determination Method Cyclodextrin Residue ⁇ 1.5% HPLC
- 1,2-propanediol hydrolysate ⁇ 2.5% GC 1 HPLC product contains only traces of glycolic acid (about 10 ⁇ 20ppm) after purification. This impurity is related to the new material of biomedical engineering, lactic acid monoglycolic acid copolymer (PLGA). The degradation products are identical. Extensive research has shown that PLGA has good biocompatibility and biodegradability, and it has good safety. It can be made into artificial catheter, drug delivery carrier, tissue engineering scaffold material, and has been rapidly developed.
- the ⁇ -cyclodextrin is liable to form a hydrogen bond, and therefore exhibits an extremely high inclusion ability for a drug having a carboxyl group and a hydroxyl group structure. Further determining the inclusion constants of the different degree of substitution products of the present invention and the above drugs, and found that the inclusion constant is about the same as the degree of substitution ( ⁇ ).
- Tests have shown that the CM-HP-p-CD prepared by the invention not only has a small self-hemolytic effect, but also can effectively reduce the hemolysis of the encapsulated drug and improve the safety of administration.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Inorganic Chemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Preparation (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
Disclosed are a carboxymethyl-hydroxypropyl-β-cyclodextrin and preparation method thereof, the carboxymethyl-hydroxypropyl-β-cyclodextrin being a β-cyclodextrin derivative mixedly substituted by carboxymethyl and hydroxypropyl, the derivative being: n-(2, 3, 6-O-carboxymethyl)-m-(2, 3, 6-O-2-hydroxypropyl)-β-cyclodextrin. The substituted groups of each mole of cyclodextrin contain n carboxymethyl and m hydroxypropyl, m being the number of connections to hydroxypropyl in each mole of cyclodextrin derivative, that is, the average degree of substitution of hydroxypropyl, and n being the number of connections to carboxymethyl in each mole of cyclodextrin derivative, that is, the average degree of substitution of carboxymethyl, where m is a number selected from 1 to 6 and n is a number selected from 1 to 6. The present invention acts as a medical excipient, and has the features of low hemolytic property, low toxicity, and strong solubility.
Description
羧甲基-羟丙基- β -环糊精及其制备方法 技术领域 本发明涉及的是羧甲基和羟丙基混合取代的用于药用辅料的 Ρ -环糊精衍生物 羧甲基-羟丙基 环糊精及其制备方法。 背景技术 TECHNICAL FIELD The present invention relates to a carboxy-methyl and hydroxypropyl mixed-substituted oxime-cyclodextrin derivative carboxymethyl group for pharmaceutical excipients. - Hydroxypropyl cyclodextrin and a process for its preparation. Background technique
β -环糊精（ β -cyclodextrin, β -CD ) 系淀粉经环糊精葡聚糖转位酶酶解环 合后得到的由 Ί 个葡萄糖分子连接而成的环状低聚糖天然产物。 分子 以糖甙键连接构成有中空内腔的截头圆锥形体，锥体空腔外围分布 21个羟基。 其腔内疏水、 腔外亲水的特性， 使得 3 -CD能与有机小分子形成主客分子弱作 用的超分子 （包合物） 。 有机小分子因形成超分子而使理化性质得到改善， 在 农业、医药业、食品及化妆品等应用领域有很高的研究和应用价值。但 P -CD 溶 解度小 （1.85% ) ， 影响了包合增溶能力， 非肠道给药具明显肾毒性， 且溶血 作用强， 只适合口服而不能用于非肠道给药。 因此， 通过 β -CD结构改造创制 新型衍生物， 提高溶解性增强包合能力、 降低溶血作用具有重要意义。 Β-cyclodextrin (β-CD) is a cyclic oligo natural product obtained by cyclization of cyclodextrin glucan transposase to obtain a cyclic oligosaccharide. The molecules are connected by a glycoside bond to form a frustoconical body having a hollow inner cavity, and 21 hydroxyl groups are distributed around the periphery of the pyramid cavity. Its hydrophobic and extracellular hydrophilic properties make it possible for 3-CD to form a supramolecular (compound) with weak organic molecules. Organic small molecules have improved physical and chemical properties due to the formation of supramolecules, and have high research and application value in agricultural, pharmaceutical, food and cosmetic applications. However, the solubility of P-CD is small (1.85%), which affects the ability of inclusion and solubilization. It has obvious nephrotoxicity in parenteral administration and has strong hemolysis effect. It is only suitable for oral administration and cannot be used for parenteral administration. Therefore, it is important to modify the β-CD structure to create new derivatives, improve the solubility and enhance the inclusion ability, and reduce the hemolysis.
药用环糊精衍生物必须具有水溶性好、 溶血性小、 毒性低， 且包合增溶能 力适当的特性。 已有研究表明， 亲水基羟丙基和磺丁基都是修饰 β-CD的良好 取代基， 由此制备的衍生物羟丙基 -β-环糊精 （ΗΡ-β-CD ) 和磺丁基 -β-环糊精 ( SBE- -CD ) 已成为目前常用的药用环糊精， 广泛用于药品和食品工业。 羧 甲基 - β -环糊精 （CM-p-CD) 已有市售产品可购买， 但目前尚没有大量工业应 用。 The pharmaceutically acceptable cyclodextrin derivative must have good water solubility, low hemolytic activity, low toxicity, and inclusion of solubilizing ability. Studies have shown that both hydrophilic hydroxypropyl and sulfobutyl groups are good substituents for the modification of β-CD, and the derivatives thus prepared are hydroxypropyl-β-cyclodextrin (ΗΡ-β-CD) and sulfonate. Butyl-β-cyclodextrin (SBE--CD) has become a commonly used medicinal cyclodextrin and is widely used in the pharmaceutical and food industries. Carboxymethyl-β-cyclodextrin (CM-p-CD) is commercially available as a commercially available product, but there are currently no large industrial applications.
目前正在使用的药用环糊精衍生物皆有高的水溶性和较强的包合性能， 其 中的甲基 -β-环糊精能溶于有机溶剂， 分子量较小 （ DS = 1〜10， M = 1149〜 nysg-moi 1 ) ， 载药效率高， 但其水溶液显强溶血作用， 刺激性强； 羟丙基 -β- 环糊精能溶于有机醇类溶剂， 分子量适中 （DS = 1〜12， M = 1193〜1831 g-mol-1 ) ， 载药效率较高， 但水溶液溶血作用明显， 刺激性较强； 磺丁基 - e - 环糊精难溶于有机溶剂， 分子量较大 （DS = 5.6〜7.4， M = 2019.8〜2304.2 g-mol"1 ) ， 载药效率较低， 但溶血作用低， 刺激性小。 The medicinal cyclodextrin derivatives currently in use have high water solubility and strong inclusion properties, wherein the methyl-β-cyclodextrin is soluble in organic solvents and has a small molecular weight (DS = 1 to 10). , M = 1149~ nysg-moi 1 ) , high drug-loading efficiency, but its aqueous solution has strong hemolysis and strong irritancy; hydroxypropyl-β-cyclodextrin is soluble in organic alcohol solvents with moderate molecular weight (DS = 1~12, M = 1193~1831 g-mol- 1 ), the drug-loading efficiency is high, but the hemolysis effect of the aqueous solution is obvious and the irritating effect is strong; the sulfobutyl-e-cyclodextrin is insoluble in the organic solvent, and the molecular weight is relatively high. Large (DS = 5.6~7.4, M = 2019.8~2304.2 g-mol" 1 ), the drug-loading efficiency is low, but the hemolytic effect is low and the irritation is small.
一般而言， 引入取代基能够使 β-CD衍生物溶解性和包合能力明显增强， 取代基类型的改变对溶解性能的影响差异不大， 而对包合能力的改变及溶血作
用的影响则较复杂， 包合能力即与取代基类型相关 （包合材料结构） ， 也与被 包合药物分子的结构和官能团类型等关系密切； 溶血作用的改变即取决于取代 基类型， 又与其取代基数量 （取代度， DS ) 相关， ΗΡ-β-CD较母体 β-CD溶血 作用有所减小， 并在较宽的 DS范围 （2-10 ) 溶血作用强弱变化不大， 而 SBE-P-CD溶血作用降低明显而且随 DS的改变波动较大， DS的增大对降低溶 血作用有利； 而羧甲基能够降低产物的溶血作用， 其 DS与溶血的关系尚不明 确， 虽然低浓度即表现明显的溶血作用， 但高浓度下具有比 HP- ø -CD更低的 溶血性。 In general, the introduction of substituents can significantly enhance the solubility and inclusion capacity of β-CD derivatives. The change in the type of substituents has little effect on the solubility properties, but the change in inclusion ability and hemolysis The impact is more complicated, and the inclusion ability is related to the type of substituent (including the structure of the material), and also closely related to the structure and functional group type of the encapsulated drug molecule; the change of hemolysis depends on the type of substituent. Also related to the number of substituents (degree of substitution, DS), ΗΡ-β-CD is less hemolytic than maternal β-CD, and has little change in hemolysis in the wider DS range (2-10). The hemolytic effect of SBE-P-CD decreased significantly and fluctuated greatly with the change of DS. The increase of DS was beneficial to reduce hemolysis. The carboxymethyl group can reduce the hemolysis of the product, and the relationship between DS and hemolysis is still unclear. Although the low concentration shows significant hemolysis, it has lower hemolytic properties than HP-ø-CD at high concentrations.
目前， 大部分的环糊精衍生物是单一取代基的衍生化产物， 近年来采用混 合取代基改进 β-CD制备质量更优的衍生物已有报道。 WO 2005042584 制备了 二元取代的磺烷基-烷基-环糊精 （SAEx-AEy-CD ) ， 不仅提高了水溶性、 增强 了包合性能， 而且其溶血性也较垸基-环糊精有明显降低， 其中乙基-磺丁基 -β- 环糊精衍生物的溶血作用与 SBE4-P-CD相当。 CN100503647C结合 ΗΡ-β-CD和 SBE-P-CD的优势， 同时以羟丙基和磺丁基取代制得羟丙基-磺丁基 -β-环糊精At present, most of the cyclodextrin derivatives are derivatized products of single substituents, and in recent years, it has been reported that mixed substituents are used to improve the quality of β-CD. WO 2005042584 Preparation of a disubstituted sulfoalkyl-alkyl-cyclodextrin (SAE x -AE y -CD ), which not only improves water solubility, enhances inclusion properties, but also has hemolytic properties compared to thiol-ring There is a significant decrease in dextrin, in which the hemolysis of the ethyl-sulfobutyl-β-cyclodextrin derivative is comparable to that of SBE4-P-CD. CN100503647C Combining the advantages of ΗΡ-β-CD and SBE-P-CD, and simultaneously preparing hydroxypropyl-sulfobutyl-β-cyclodextrin with hydroxypropyl and sulfobutyl groups
(HP-SBE-P-CD ) ， 能够获得较低溶血性的衍生物产品， 显示了较好的应用前 景。 但其较高分子量导致的载药率偏低、 过高的制剂辅料用量明显制约了产 品的实际应用。 (HP-SBE-P-CD), a derivative product that achieves lower hemolysis, shows a better application prospect. However, the higher drug-loading rate caused by the higher molecular weight and the excessive use of the auxiliary materials significantly restrict the practical application of the product.
除满足水溶性、 包合性能和安全性要求之外， 药用环糊精衍生物的分子量 也是实际应用中必须考虑的重要因素。 作为药物载体， 过大的分子量将使处方 辅料用量过高， 而影响环糊精产品实际作用功能的发挥， 成为限制其实用性的 重要原因。 在确保水溶性和安全性基础上， 设计制备较低分子量的环糊精衍生 物不仅具有重要意义， 也是目前药用环糊精发展的技术难点之一。 发明内容 In addition to meeting water solubility, inclusion properties and safety requirements, the molecular weight of pharmaceutically acceptable cyclodextrin derivatives is also an important factor that must be considered in practical applications. As a drug carrier, an excessively large molecular weight will make the amount of prescription excipients too high, and the effect of the actual function of the cyclodextrin product becomes an important reason for limiting its practicability. On the basis of ensuring water solubility and safety, the design and preparation of lower molecular weight cyclodextrin derivatives is not only of great significance, but also one of the technical difficulties in the development of pharmaceutical cyclodextrins. Summary of the invention
本发明拟解决的技术问题： 改善 β-环糊精水溶性， 制备出安全、 有效而又 具有相对较低分子量的 β-环糊精衍生物。 其技术方法是， 在 β-环糊精母体中同 时引入两种较小的亲水取代基， 以达到改善 β-环糊精性能、 提高产品安全性、 增强产品实用性的目的。 本发明的技术解决方案是： The technical problem to be solved by the present invention is to improve the water solubility of β-cyclodextrin and to prepare a β-cyclodextrin derivative which is safe, effective and has a relatively low molecular weight. The technical method is to introduce two smaller hydrophilic substituents in the β-cyclodextrin precursor to improve the performance of β-cyclodextrin, improve product safety, and enhance product practicability. The technical solution of the present invention is:
设计一种以羧甲基和羟丙基混合取代的 β-环糊精衍生物 η-(2,3，6-Ο-羧甲 基) -m-(2,3，6-O-2-羟丙基) -β-环糊精 （简称：羧甲基-羟丙基 - β -环糊精） ， 该 β-环 糊精衍生物具有下列通式所示的结构：
Rl=H R CHiCOO-X, ¾= CH2CH(OI¾CHj , ¾ Design of a β-cyclodextrin derivative η-(2,3,6-fluorenyl-carboxymethyl)-m-(2,3,6-O-2-) substituted with a mixture of carboxymethyl and hydroxypropyl groups Hydroxypropyl)-β-cyclodextrin (abbreviation: carboxymethyl-hydroxypropyl-β-cyclodextrin), the β-cyclodextrin derivative having the structure shown by the following formula: Rl=HR CHiCOO-X, 3⁄4= CH 2 CH(OI3⁄4CHj , 3⁄4
本发明中所述的羧甲基-羟丙基 -e-环糊精， 包含多种上述分子结构的化合 物。 上述结构式中，
CH2CH(OH)CH3、 H" ， 是指当 R^OR 均为 CH2COO-X时， 上述结构式中的 7个 R3基团中， 部分 R3基团为 CH2CH(OH)CH3, 另一部分 R3基团为 H。 其余类似表述均为此意。 羧甲基-羟丙基- β -环糊精可以 CMn-HPm-p-CD表示， 其结构中羧甲基 (-CH2COO-X) 和 2-羟丙基 （-CH2CH(OH)CH3) 是同时连接于环糊精母体形 成的醚键取代衍生物， 即该环糊精衍生物同时具有羧甲基和羟丙基两种取代基 团。 每摩尔 β-环糊精连接的取代基平均数即为平均取代度 （DS) 有： η个羧甲基 和 m个羟丙基。 其中 m为羟丙基平均取代度； n为羧甲基平均取代度， m + n - Z 为总平均取代度； 混合取代产物必须 n l及 m l。 其产物的羟丙基或羧甲 基的取代位是随机取代于 环糊精的 2位， 或 3位， 或 6位的衍生物。 羧甲基 和羟丙基混合取代的环糊精衍生物（简称：羧甲基-羟丙基 -e-环糊精）， 不是羟丙基 - -环糊精与羧甲基- β -环糊精二者的简单混合物。 羧甲基-羟丙基 -β-环糊精中， 羧甲基的平均取代度范围是 1〜6， 即 η= 1、The carboxymethyl-hydroxypropyl-e-cyclodextrin described in the present invention contains a plurality of compounds of the above molecular structure. In the above structural formula, CH 2 CH(OH)CH 3 , H" , means that when R^OR is CH 2 COO-X, among the 7 R 3 groups in the above structural formula, part of the R 3 group is CH 2 CH(OH CH 3 , another part of the R 3 group is H. The rest of the similar expressions are all this. Carboxymethyl-hydroxypropyl-β-cyclodextrin can be represented by CMn-HPm-p-CD, the structure of which is carboxymethyl (-CH2COO-X) and 2-hydroxypropyl (-CH 2 CH(OH)CH 3 ) are ether bond-substituted derivatives formed by simultaneous attachment to a cyclodextrin precursor, ie, the cyclodextrin derivative has both a carboxyl group Base and hydroxypropyl two substituent groups. The average number of substituents per mole of β-cyclodextrin is the average degree of substitution (DS): η carboxymethyl and m hydroxypropyl. Where m is hydroxy The average degree of substitution of propyl; n is the average degree of substitution of carboxymethyl groups, m + n - Z is the total average degree of substitution; the mixed substituted product must be nl and ml. The substitution position of the hydroxypropyl or carboxymethyl group of the product is randomly substituted. Derivative of cyclodextrin at position 2, or position 3 or position 6. Carboxymethyl and hydroxypropyl mixed substituted cyclodextrin derivatives (abbreviation: carboxymethyl-hydroxypropyl-e-cyclodextrin ), not hydroxypropyl-cyclodextrin and carboxymethyl- a simple mixture of both β-cyclodextrin. In carboxymethyl-hydroxypropyl-β-cyclodextrin, the average degree of substitution of carboxymethyl groups ranges from 1 to 6, ie η = 1,
2、 3、 4、 5和 6中的任一值； 羟丙基的平均取代度范围是 1〜6， 即 m= l、 2、2, 3, 5, and 6; the average degree of substitution of the hydroxypropyl group is 1 to 6, that is, m = 1, 2
3、 4、 5和 6中的任一值； 而两取代基加合的总平均取代度 Z = 2、 3、 4、 5、 6、 7、 8、 9和 10中的任一值。 如：
-羧甲基-一 -羟丙基- β -环糊精 (CMi-HPi-•β-■CD Z = 2) 、Any of the values of 3, 4, 5, and 6; and the total average degree of substitution of the two substituents Z = 2, 3, 4, 5, 6, 7, 8, 9, and 10. Such as: -carboxymethyl-mono-hydroxypropyl-β-cyclodextrin (CMi-HPi-•β-■CD Z = 2),
-羧甲基-二 -羟丙基- -环糊精 (CMi-HP2- β· ■CD Z = 3) 、 - Carboxymethyl - di - hydroxypropyl - - cyclodextrin (CMi-HP 2 - β · ■ CD Z = 3),
-羧甲基-二 -羟丙基- β -环糊精 (CM2-HP2- •β. ■CD Z = 4) 、 -carboxymethyl-di-hydroxypropyl-β-cyclodextrin (CM 2 -HP 2 - •β. ■CD Z = 4),
二 -羧甲基-二 -羟丙基- β -环糊精 (CM3-HP2- β- ■CD Z = 5) 、 Di-carboxymethyl-di-hydroxypropyl-β-cyclodextrin (CM 3 -HP 2 - β- ■CD Z = 5),
四 -羧甲基-一 -羟丙基- -环糊精 (CM4-HPi- β· ■CD Z = 5) 、 Tetra-carboxymethyl-mono-hydroxypropyl-cyclodextrin (CM 4 -HPi-β· ■CD Z = 5),
-羧甲基-三 -羟丙基- β -环糊精 (CM2-HP3- •β. ■CD Z = 5) 、 -carboxymethyl-tris-hydroxypropyl-β-cyclodextrin (CM 2 -HP 3 - •β. ■CD Z = 5),
-羧甲基-四 -羟丙基- β -环糊精 (CM2-HP4- β· ■CD Z = 6) 、 -carboxymethyl-tetra-hydroxypropyl-β-cyclodextrin (CM 2 -HP 4 - β· ■ CD Z = 6),
二 -羧甲基-三 -羟丙基- β -环糊精 (CM3-HP3- •β. ■CD Z = 6) 、 Di-carboxymethyl-tris-hydroxypropyl-β-cyclodextrin (CM3-HP3- •β. ■CD Z = 6),
二 -羧甲基-四 -羟丙基- β -环糊精 (CM3-HP4- ·β· ■CD Z = 7) 、 Di-carboxymethyl-tetra-hydroxypropyl-β-cyclodextrin (CM 3 -HP 4 - ·β· ■CD Z = 7),
-羧甲基-五 -羟丙基- β -环糊精 (CM2-HP5- ■β. ■CD Z = 7) 、 -carboxymethyl-penta-hydroxypropyl-β-cyclodextrin (CM 2 -HP 5 - ■β. ■CD Z = 7),
-羧甲基-五 -羟丙基- β -环糊精 (CM3-HP5- •β' ■CD Z = 8) 、 -carboxymethyl-penta-hydroxypropyl-β-cyclodextrin (CM 3 -HP 5 - •β' ■CD Z = 8),
~ - -羧甲基-六 -轻丙基- β -环糊精 (CM2-HP6- •β. ■CD Z = 8) 、 ~ - -Carboxymethyl-hexa-propylpropyl-β-cyclodextrin (CM 2 -HP 6 - •β. ■CD Z = 8),
_、 . _, .
/、 -羧甲基-三 -羟丙基- β -环糊精 (CM6-HP3- ■β- ■CD Z = 9) 、 /, -Carboxymethyl-tris-hydroxypropyl-β-cyclodextrin (CM 6 -HP 3 - ■β- ■CD Z = 9),
五 -羧甲基-五 -羟丙基- -环糊精 (CM5-HP5- β· ■CD Z = 10) 。 Penta-carboxymethyl-penta-hydroxypropyl-cyclodextrin (CM5-HP5-β· ■ CD Z = 10).
由环糊精衍生物结构特点表示的平均取代度 n 和 tn还包含其 ±0.5内的实 际平均值， 各可选衍生物的平均取代度范围是 The average degree of substitution n and tn represented by the structural characteristics of the cyclodextrin derivative also contain the actual average value within ±0.5, and the average degree of substitution for each of the optional derivatives is
CM3-HP2- -CD n = 2.5〜3.4; m： 1.5〜2·4、CM 3 -HP 2 - -CD n = 2.5~3.4; m: 1.5~2·4,
CM4-HPi- -CD n = 3.5〜4.4; m = 1.0—1.4. CM 4 -HPi- -CD n = 3.5~4.4; m = 1.0-1.4.
CM6-HP3-p-CD n = 5.5-6.4； m = 2.5〜3.4、CM 6 -HP 3 -p-CD n = 5.5-6.4; m = 2.5~3.4,
CM2-HP2-P-CD n = 1.5-2.4； m = 1.5〜2.4、CM 2 -HP 2 -P-CD n = 1.5-2.4; m = 1.5~2.4,
CM4-HP3- -CD n = 3.5-4.4； m = 2.5〜3.4、CM 4 -HP 3 - -CD n = 3.5-4.4; m = 2.5~3.4,
CM2-HP4-P-CD n = 1.5-2.4； m = 3.5〜4.4、 CM 2 -HP 4 -P-CD n = 1.5-2.4; m = 3.5~4.4,
CM3-HP4-p-CD n = 2.5〜3.4; m = 3.5〜4.4、CM 3 -HP 4 -p-CD n = 2.5~3.4; m = 3.5~4.4,
CM2-HP5-p-CD n = 1.5-2.4； m = 4.5〜5·4、CM 2 -HP 5 -p-CD n = 1.5-2.4; m = 4.5~5·4,
CM3-HP5-p-CD n = 2.5-3.4； m = 4.5〜5.4、CM 3 -HP 5 -p-CD n = 2.5-3.4; m = 4.5~5.4,
CM2-HP6-p-CD n = 1.5〜2.4; m = 5.5— 6.4或CM 2 -HP 6 -p-CD n = 1.5~2.4; m = 5.5- 6.4 or
CM2-HP3-p-CD n = 1·5〜2·4; m = 2·5〜3.4。 本发明所设计的羧甲基-羟丙基- β -环糊精中， 优选 Ζ = 4〜7的如下结构
CM.-HP3-P-CD (n = 1， m =3, Z = 4) CM 2 -HP 3 -p-CD n = 1 · 5~2 · 4; m = 2 · 5~3.4. In the carboxymethyl-hydroxypropyl-β-cyclodextrin designed by the present invention, the following structure of Ζ = 4 to 7 is preferred. CM.-HP 3 -P-CD (n = 1, m = 3, Z = 4)
CM2-HP2- -CD (n =2， m =2， Z = 4) CM 2 -HP 2 - -CD (n = 2, m = 2, Z = 4)
CM1-HP4- -CD (n = 1, m =4, z = 5) CM 1 -HP 4 - -CD (n = 1, m = 4, z = 5)
CM2-HP3- -CD (n =2, m =3, z = 5) CM 2 -HP 3 - -CD (n = 2, m = 3, z = 5)
CM3-HP2-p-CD (n =3， m =2, z = 5) CM 3 -HP 2 -p-CD (n =3, m =2, z = 5)
CMi-HP5-p-CD (n = 1， m = 5， z = 6) CMi-HP 5 -p-CD (n = 1, m = 5, z = 6)
CM2-HP4-P-CD (n =2, m =4， z = 6) CM 2 -HP 4 -P-CD (n =2, m =4, z = 6)
CM3-HP3-p-CD (n =3， m =3, z = 6) CM 3 -HP 3 -p-CD (n =3, m =3, z = 6)
CMj-HPe- -CD (n = 1， m =6, z = 7) CMj-HPe- -CD (n = 1, m =6, z = 7)
CM2-HP5-p-CD (n =2， m = 5, z = 7) CM 2 -HP 5 -p-CD (n =2, m = 5, z = 7)
CM3-HP4- -CD (n =3, m =4, z = 7) 本发明所设计的羧甲基-羟丙基 -e-环糊精以 β-环糊精、 氯乙酸乙酯和 1,2- 环氧丙垸为原料制备， 在水中加入 β-环糊精及催化量碱， 在适当温度搅拌下先 后滴加氯乙酸乙酯和 1,2-环氧丙垸反应， 再经过纯化处理制得系列设计物 CMn-HPm-P-CDo 以此方法， 调整试剂及催化剂碱的不同投料比即可制得设计 所需取代度的产物： 随试剂投料比的增加， 产物取代度即不断提高， 降低试剂 投料比取代度亦减小； 试剂投料比氯乙酸乙酯 2.0倍摩尔量 （相对 环糊精） 以上、 1,2-环氧丙烷最低 1.5倍摩尔量以上能制得 n l及 m^l的混合取代产物； 7倍 环糊精摩尔量加 2倍氯乙酸乙酯试剂的投料量 （摩尔） 为催化剂碱的总 投料量。 以 NaOH催化， 可制备得 X = Na、 H的产物， KOH催化则得 X = K、 Η的产物。 CM 3 -HP 4 - -CD (n = 3, m = 4, z = 7) The carboxymethyl-hydroxypropyl-e-cyclodextrin designed by the present invention is β-cyclodextrin, ethyl chloroacetate It is prepared by using 1,2-epoxypropionam as a raw material, adding β-cyclodextrin and a catalytic amount of alkali in water, and adding ethyl chloroacetate and 1,2-epoxypropanoid dropwise after stirring at an appropriate temperature. After purification, a series of designs CMn-HPm-P-CDo can be obtained. By adjusting the different ratios of reagents and catalyst bases, the product with the desired degree of substitution can be obtained: with the increase of the reagent ratio, the degree of product substitution. That is to say, it is continuously improved, and the ratio of substitution of reagents is also reduced. The reagent charge can be obtained by adding 2.0 times the molar amount of ethyl chloroacetate (relative to cyclodextrin) and 1.5 times or more by mole of 1,2-epoxypropane. And a mixed substitution product of m^l; a molar amount of 7 times cyclodextrin plus 2 times of ethyl chloroacetate reagent (molar) is the total amount of catalyst base. The product of X = Na and H can be prepared by catalysis with NaOH, and the product of X = K and hydrazine can be obtained by KOH catalysis.
进一步地， 本发明提供了一种上述的羧甲基-羟丙基- β -环糊精的制备方 法，采用碱催化连续反应的方法进行制备，具体反应步骤是以下两种方案之一： 方案 1: 取 β-环糊精加入 0-环糊精 2.2倍质量的水和 7.0倍摩尔量的碱，搅 拌溶解， 室温下搅拌滴加 7.0倍摩尔量的 1，2-环氧丙烷反应， 数小时后补加 8.0 倍摩尔量的碱， 加热， 升温稳定后加入 4.0摩尔量的氯乙酸乙酯， 反应生成特 定取代度的羧甲基-羟丙基- β -环糊精产物；
方案 2: 取 β -环糊精加入 P -环糊精 2.2倍质量的水和 11.0~35.0倍摩尔量 的碱， 搅拌溶解， 加热下滴加 2.0~10.5倍摩尔量的氯乙酸乙酯反应数小时， 冷 却至室温， 然后控制室温下加入 1.5〜7.0倍摩尔量的 1,2-环氧丙烷， 反应数小时 即生成特定取代度的羧甲基-羟丙基 - β -环糊精产物。 Further, the present invention provides a preparation method of the above carboxymethyl-hydroxypropyl-β-cyclodextrin, which is prepared by a method of base-catalyzed continuous reaction, and the specific reaction step is one of the following two schemes: 1: Take β-cyclodextrin and add 2.2 times mass of water and 7.0 times molar amount of base to 0-cyclodextrin, stir to dissolve, and add 7.0 times molar amount of 1,2-epoxypropane to react at room temperature. After an hour, 8.0 times the molar amount of the base is added, heated, and the temperature is stabilized, and 4.0 molar amount of ethyl chloroacetate is added to react to form a carboxymethyl-hydroxypropyl-β-cyclodextrin product having a specific degree of substitution; Scheme 2: Add β-cyclodextrin to P-cyclodextrin 2.2 times the mass of water and 11.0~35.0 times the molar amount of alkali, stir to dissolve, and add 2.0~10.5 times molar amount of ethyl chloroacetate reaction under heating. After cooling to room temperature, 1.5 to 7.0 times the molar amount of 1,2-propylene oxide was added at room temperature, and a specific degree of substitution of the carboxymethyl-hydroxypropyl-β-cyclodextrin product was formed in a few hours.
进一步地， 本发明中以母体 环糊精为原料， 采用反应中间体不经分离的 连续加料反应步骤进行混合取代。 Further, in the present invention, the parent cyclodextrin is used as a raw material, and the reaction intermediate is mixed and replaced by a continuous feeding reaction step without separation.
进一步地， 本发明提供了上述的任一羧甲基-羟丙基 环糊精在药学上的 应用， 包括所述的羧甲基-羟丙基 - β -环糊精在药学上作为赋形剂应用于包合活 性分子制备的组合物。 Further, the present invention provides the pharmaceutical use of any of the above carboxymethyl-hydroxypropyl cyclodextrin, comprising the pharmaceutically acceptable form of carboxymethyl-hydroxypropyl-β-cyclodextrin The agent is applied to a composition comprising an active molecule preparation.
进一步地， 本发明提供了上述的该含有所述羧甲基-羟丙基 - 3 -环糊精的组 合物用作治疗或保健用途的口服制剂产品。 Further, the present invention provides the above oral preparation product containing the carboxymethyl-hydroxypropyl-3-cyclodextrin composition for therapeutic or health care use.
进一步地， 本发明提供了上述羧甲基-羟丙基 环糊精的组合物用作治疗 或保健用途的非口服制剂产品。 Further, the present invention provides a composition of the above carboxymethyl-hydroxypropyl cyclodextrin for use as a non-oral preparation product for therapeutic or health care purposes.
迸一步地， 本发明提供了上述的任一羧甲基-羟丙基 环糊精作为药物辅 料的应用。 本发明的优点 Further, the present invention provides the use of any of the above carboxymethyl-hydroxypropyl cyclodextrin as a pharmaceutical adjuvant. Advantages of the invention
1、 本发明合成的羧甲基-羟丙基 -β-环糊精溶血作用显著低于相应的单取代 基衍生物 ΗΡ-β-CD和 CM-p-CD， 溶血率低， 基本无毒性。 无有害残留物， 产 品安全性强。 不仅产物自身溶血作用小， 还能够明显降低被包合药物溶血率， 提高用药安全性。 1. The hemolytic effect of the synthesized carboxymethyl-hydroxypropyl-β-cyclodextrin of the invention is significantly lower than that of the corresponding monosubstituted derivatives ΗΡ-β-CD and CM-p-CD, with low hemolysis rate and substantially no toxicity. . No harmful residues, the product is safe. Not only the product itself has a small hemolysis effect, but also can significantly reduce the hemolysis rate of the encapsulated drug and improve the safety of administration.
2、 产品溶解性优异， 水中溶解度 50%， 有机溶解剂中也表现出良好的溶 解性。 为包合技术改进难溶药物溶解性奠定了物质基础， 在难溶药物制剂的应 用将有广阔前景。 2. The product has excellent solubility, solubility in water is 50%, and organic solvent also shows good solubility. It has laid a material foundation for improving the solubility of poorly soluble drugs by inclusion technology, and has a broad prospect in the application of poorly soluble pharmaceutical preparations.
3、 本发明的产品包合药物能力适当， 对各类型药物 （包括多肽大分子） 皆有良好的包合性能。 对提高药物溶解度、 增强吸收和改善生物利用度具有重 要应用价值。 3. The product of the invention has the appropriate ability to contain drugs, and has good inclusion properties for various types of drugs, including polypeptide macromolecules. It has important application value for improving drug solubility, enhancing absorption and improving bioavailability.
4、 本发明制备的羧甲基-羟丙基 -β-环糊精具有适度的平均分子量， 与现有 产品 ΗΡ-β-CD分子量相当， 但溶血作用显著低于 ΗΡ-β-CD, 从而极大的提高了 本发明产品的应用可行性。 4. The carboxymethyl-hydroxypropyl-β-cyclodextrin prepared by the invention has a moderate average molecular weight, which is equivalent to the molecular weight of the existing product ΗΡ-β-CD, but the hemolytic effect is significantly lower than that of ΗΡ-β-CD, thereby The application feasibility of the product of the invention is greatly improved.
5、 本发明的制备和分析方法简单， 制备产率高， 质量稳定。 系列产物的 制备重量收率 100%。 另外， 本发明制备方法原料易得 （市售 p-CD、 1,2-环氧 丙烷、 氯乙酸乙酯） ， 生产成本低。 制备过程中通过调节投料比就可以得到所
需取代度的产品。 5. The preparation and analysis method of the invention is simple, the preparation yield is high, and the quality is stable. The product of the series was prepared in a weight yield of 100%. Further, the preparation method of the present invention is easy to obtain (commercially available p-CD, 1,2-propylene oxide, ethyl chloroacetate), and the production cost is low. In the preparation process, the feed ratio can be adjusted to obtain the Products that need to be replaced.
6、 本发明产品性质稳定， 无毒害无易燃易爆特性、 无环境污染， 不易变 质， 易储藏、 易运输。 附图说明 图 1 : 产物 HSQC谱 图 2: 产物 COSY谱 图 3: 产物 ROESY谱 图 4: 产物 13CNMR谱 图 5: 产物！HNMR谱 6. The product of the invention has stable properties, no toxicity, no flammable and explosive characteristics, no environmental pollution, no deterioration, easy storage and easy transportation. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1: Product HSQC spectrum Figure 2: Product COSY spectrum Figure 3: Product ROESY spectrum Figure 4: Product 13 CNMR spectrum Figure 5: Product! HNMR spectrum
图 6: 产物差示热分析图: a: β-CD; b: ΗΡ-β-CD; c: CM-P-CD; d: HP-p-CD/p-CD/CM-p-CD三者混合 物； e: CM2-HP6- -CD 图 7: 产物 HPLC色谱 图 8: p-CD、 CM-p-CD和 ΗΡ-β-CD的三者物理混合物 HPLC色谱 图 9: 青蒿素、 CM3-HP6-p-CD、 青蒿素与 CM3-HP6-p-CD物理混合物、 CM3-HP6-P-CD/青蒿素包合物四样品对比 DTA图谱： a:青蒿素； b: CM3-HP6-p-CD; c:青蒿素 /CM3-HP6-p_CD物理混合物； d: CM3-HP6-P-CD/青蒿素包合物 附图 10: 多种环糊精的体外胰岛素转运率 Figure 6: Product differential thermogram: a: β-CD; b: ΗΡ-β-CD; c: CM-P-CD; d: HP-p-CD/p-CD/CM-p-CD E; CM 2 -HP 6 - -CD Figure 7: HPLC chromatogram of product Figure 8: HPLC chromatogram of three physical mixtures of p-CD, CM-p-CD and ΗΡ-β-CD Figure 9: Artemisinin , CM 3 -HP 6 -p-CD, artemisinin and CM 3 -HP 6 -p-CD physical mixture, CM 3 -HP 6 -P-CD / artemisinin inclusion complex four samples comparison DTA map: a : artemisinin; b: CM 3 -HP 6 -p-CD ; c: artemisinin / CM 3 -HP 6 -p_CD physical mixture; d: CM 3 -HP 6 -P-CD / artemisinin inclusion Figure 10: In vitro insulin transport rate of various cyclodextrins
具体实施方式 detailed description
实施例 1 Example 1
三口圆底烧瓶装配恒压漏斗、 回流冷凝管、 温度计， 向其中加入 50ml纯 净水， 开动搅拌， 再依次搅拌下缓缓加入 0.02πιο1 β -环糊精 （22.7g)和 0.3mol 固体 NaOH ( 12.0g)， 体系溶解后缓慢加热， 保持 80°C， 然后缓慢滴加 0.08mol 氯乙酸乙酯 （9.8g) , 约 4h加完， 继续搅拌回流反应 2h后冷却至室温。 水浴
控制温度<20 , 搅拌下向体系中缓慢滴加 0.14mol的 1，2-环氧丙垸 （8.1g) ， 约 3h加完， 水浴温度<20 条件下继续搅拌反应 5h， 加入盐酸调 pH至中性， 过滤， 滤液倒入透析袋中， 透析除去残留的 环糊精和反应生成的 1，2-丙二醇 及羟基乙酸 （钠） 。 换水透析 10次， 袋内溶液<60°( 下减压浓縮， 再 60Ό下减 压干燥， 得 X = Na、 H的类白色固体产物 25.0g， 产率 110.1%。 A three-neck round bottom flask was equipped with a constant pressure funnel, a reflux condenser, and a thermometer. 50 ml of purified water was added thereto, and stirring was started. Then, 0.02πιο1 β-cyclodextrin (22.7 g) and 0.3 mol of solid NaOH (12.0) were slowly added thereto with stirring. g), after the system is dissolved, slowly heat, keep 80 ° C, then slowly add 0.08 mol of ethyl chloroacetate (9.8 g), add about 4 h, continue to stir and reflux for 2 h and then cool to room temperature. Water bath Control the temperature <20, slowly add 0.14mol of 1,2-epoxypropionamidine (8.1g) to the system under stirring, add about 3h, stir the reaction for 5h under the water bath temperature <20, add hydrochloric acid to adjust the pH to Neutral, filtered, the filtrate was poured into a dialysis bag, and the residual cyclodextrin and the 1,2-propanediol and glycolic acid (sodium) formed by the reaction were removed by dialysis. The solution was dialyzed 10 times with water, and the solution in the bag was <60° (concentrated under reduced pressure, and dried under reduced pressure at 60 Torr to obtain 25.0 g of a white solid product of X = Na, H, yield 110.1%.
以 D2O为溶剂， 测定产物 iHNMR谱（附图 5 ) ， 以 δ =5.07峰面积积分标 记为 7， 其 S =5.28的亚甲基单峰面积 （5.41 ) 的 1/2值即为 CM基取代度 （n = 2.70 ) ， 其 δ =1.13〜1.15的甲基二重峰面积 （17.66 ) 值 1/3即为 HP基取代度 (m = 5.89 ) ， 因此， 合成产物简写为： CM3-HP6-P-CD The product iH NMR spectrum (Fig. 5) was determined by using D 2 O as a solvent, and the integral of δ = 5.07 peak area was 7 , and the 1/2 value of the methylene single peak area (5.41 ) of S = 5.28 was CM. The degree of substitution (n = 2.70), the area of the methyl doublet of δ = 1.13 to 1.15 (17.66), and the value of 1/3 is the degree of HP substitution (m = 5.89). Therefore, the synthesized product is abbreviated as: CM 3 -HP 6 -P-CD
IR (KB r ) if : 2969cm-1 , 1461cm 1 , 1420cm'1和 1617cm-1 IR (KB r ) if : 2969cm- 1 , 1461cm 1 , 1420cm' 1 and 1617cm- 1
DTA谱： 95 'C脱水吸热峰， 270°C熔点， 300〜335°C放热分解。 DTA spectrum: 95 'C dehydration endothermic peak, 270 ° C melting point, 300 ~ 335 ° C exothermic decomposition.
理化参数如下： 分子量 M- MSS^g'mol-1 ; 含水量： 5.1% (参考： 2000 年药典附录 VMM水分测定法-甲苯法） ； 比旋光度: [ α ]=+85.98 (参考： 旋光度 测定法 2000年药典附录 VIE旋光度测定法） 。 The physical and chemical parameters are as follows: Molecular weight M-MSS^g'mol- 1 ; Water content: 5.1% (Reference: 2000 Pharmacopoeia Appendix VMM Moisture Determination - Toluene method); Specific optical rotation: [α]=+85.98 (Reference: Optical rotation Determination of the 2000 Pharmacopoeia Appendix VIE optical rotation method).
纯度测定： 残留 β -环糊精 0.82% (HPLC法）， 1,2-丙二醇 1.1% ( GC法）， 羟基乙酸<20 1!1 (沉淀法） 。 Purity determination: Residual β-cyclodextrin 0.82% (HPLC method), 1,2-propanediol 1.1% (GC method), glycolic acid <20 1!1 (precipitation method).
实施例 2 Example 2
向装有恒压漏斗、 回流冷凝管、 温度计的三口圆底烧瓶中加入 50ml纯净 水，开动搅拌，再依次缓缓加入 0.02mol β -环糊精（22.7g)和 0.14mol固体 NaOH ( 12.0g )，体系溶解后，水浴控制温度 <20°C，搅拌下向体系中缓慢滴加 0.14mol 的 1,2-环氧丙垸 （8.1g) ， 约 3h加完， 水浴温度<20 条件下继续搅拌反应 5h 后补加 0.16mol的 NaOH, 再缓慢加热， 保持 80°C， 然后缓慢滴加 0.08mol氯 乙酸乙酯 （9.8g) ， 约 4h加完， 继续搅拌回流反应 2h后冷却至室温。 加入盐 酸调 pH至中性， 过滤， 滤液倒入透析袋中， 透析除去残留的 β -环糊精和反应 生成的 1,2-丙二醇及羟基乙酸 （钠） 。 换水透析 10次， 袋内溶液<601：下减压 浓缩，再 60°C下减压干燥，得 X = Na、H的类白色固体产物 20.5g，产率 90.3%。 'HNMR验证羧甲基平均取代数度 2.1， 羟丙基的平均取代度 5.8， 产物简写为 CM2-HP6-p-CD。 Add 50 ml of purified water to a three-neck round bottom flask equipped with a constant pressure funnel, a reflux condenser, and a thermometer. Stir and add 0.02 mol of β-cyclodextrin (22.7 g) and 0.14 mol of solid NaOH (12.0 g). After the system is dissolved, the temperature of the water bath is controlled to <20 ° C, and 0.14 mol of 1,2-epoxypropionamidine (8.1 g) is slowly added dropwise to the system under stirring, and the addition is completed in about 3 hours, and the water bath temperature is <20. After stirring for 5 h, 0.16 mol of NaOH was added, and the mixture was slowly heated to maintain 80 ° C. Then, 0.08 mol of ethyl chloroacetate (9.8 g) was slowly added dropwise thereto, and the addition was completed for about 4 hours. The mixture was further stirred and refluxed for 2 hours, and then cooled to room temperature. Hydrochloric acid was added to adjust the pH to neutrality, and the filtrate was poured into a dialysis bag, and the residual β-cyclodextrin and the 1,2-propanediol and glycolic acid (sodium) formed by the reaction were dialyzed off. The mixture was dialyzed for 10 times, and the solution in the bag was <601: concentrated under reduced pressure, and dried under reduced pressure at 60 ° C to obtain 20.5 g of a white solid product of X = Na, H, yield 90.3%. 'HNMR verified that the average number of substitutions of the carboxymethyl group was 2.1, and the average degree of substitution of the hydroxypropyl group was 5.8. The product was abbreviated as CM 2 -HP 6 -p-CD.
实施例 3 Example 3
与实施例 1基本相同， 但是其中 NaOH、 氯乙酸乙酯、 1，2-环氧丙垸的投料 量分别是 0.22、 0.04、 0.05mol。 最后得产品产率为 98.5%， 经 1 HNMR验证羧 甲基平均取代度 1.2， 羟丙基平均取代度 1.8， 产物简写为 CM!-HPz-p-CDa
实施例 4 It was basically the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.22, 0.04, and 0.05 mol, respectively. The final product yield was 98.5%. The average degree of substitution of carboxymethyl group was 1.2 by 1 H NMR, and the average degree of substitution of hydroxypropyl group was 1.8. The product was abbreviated as CM!-HPz-p-CDa. Example 4
与实施例 1基本相同， 但是其中 NaOH、 氯乙酸乙酯、 1，2-环氧丙垸的投料 量分别是 0.22、 0.04、 0.08mol。 最后得产品产率为 99.1%， 经 1 HNMR验证羧 甲基平均取代度 1.1， 羟丙基平均取代度 3.4， 产物简写为 CM!-HPHi-CDa 实施例 5 Basically the same as in Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.22, 0.04, and 0.08 mol, respectively. The yield of the final product was 99.1%. The average degree of substitution of the carboxymethyl group was 1.1 by the 1 H NMR, and the average degree of substitution of the hydroxypropyl group was 3.4. The product was abbreviated as CM!-HPHi-CDa.
与实施例 1基本相同， 但是其中 NaOH、 氯乙酸乙酯、 1,2-环氧丙垸的投料 量分别是 0.26、 0.06、 0.06mol。 最后得产品产率为 100.7%， 经 1 HNMR验证羧 甲基平均取代度 1.9， 羟丙基平均取代度 2.2， 产物简写为 CM2-HPH3-CD。 The same as Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.26, 0.06, and 0.06 mol, respectively. The yield of the final product was 100.7%. The average degree of substitution of carboxymethyl group was 1.9 by 1 H NMR, and the average degree of substitution of hydroxypropyl group was 2.2. The product was abbreviated as CM 2 -HPH3-CD.
实施例 6 Example 6
与实施例 1基本相同， 但是其中 NaOH换为 KOH， 其 KOH、 氯乙酸乙酯、 1,2-环氧丙烷的投料量分别是 0.22、 0.04、 0.09mol。最后得钾盐的产物 （X = K、 Η) ， 产率为 106.8%， 经 1 HNMR验证羧甲基平均取代度 1.2， 羟丙基平均取代 度 4.1， 产物简写为 CM!-HP^p-CD a Basically the same as in Example 1, except that NaOH was changed to KOH, and the amounts of KOH, ethyl chloroacetate, and 1,2-propylene oxide were 0.22, 0.04, and 0.09 mol, respectively. Finally, the product of potassium salt (X = K, Η), the yield was 106.8%, the average degree of substitution of carboxymethyl group was 1.2 by 1 H NMR, the average degree of substitution of hydroxypropyl group was 4.1, and the product was abbreviated as CM!-HP^p- CD a
实施例 Ί Example Ί
与实施例 1基本相同， 但是其中 NaOH换为 KOH, 其 KOH、 氯乙酸乙酯、 1,2-环氧丙垸的投料量分别是 0.34、 0.1、 0.06mol。 最后得钾盐的产物 （X = K、 Η) , 产率为 111.2%， 经 1 HNMR验证羧甲基平均取代度 3.3， 羟丙基平均取代 度 2.4， 产物简写为 CM3-HP2-p-CD。 实施例 8 Basically the same as in Example 1, except that NaOH was changed to KOH, and the amounts of KOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.34, 0.1, and 0.06 mol, respectively. Finally, the product of potassium salt (X = K, Η), the yield was 111.2%, the average degree of substitution of carboxymethyl group was 3.3 by 1 H NMR, the average degree of substitution of hydroxypropyl group was 2.4, and the product was abbreviated as CM 3 -HP 2 -p -CD. Example 8
与实施例 1基本相同， 但是其中 NaOH、 氯乙酸乙酯、 1，2-环氧丙垸的投料 量分别是 0.22、 0.04、 0.12mol。 最后得产品产率为 106.2%， 经 1 HNMR验证羧 甲基平均取代度 1.3， 羟丙基平均取代度 4.6， 产物简写为 CMi-HPs-P-CDn 实施例 9 与实施例 1基本相同， 但是其中 NaOH换为 KOH， 其 KOH、 氯乙酸乙酯、 1,2-环氧丙烷的投料量分别是 0.33、 0.09、 0.07mol o最后得钾盐的产物 （X = K、 Η) ， 产率为 110.9%， 经 1 HNMR验证羧甲基平均取代度 2.9， 羟丙基平均取代 度 2.8， 产物简写为 CM3-HP3-P-CD。 实施例 10 与实施例 1基本相同， 但是其中 NaOH、 氯乙酸乙酯、 1,2-环氧丙烷的投料 量分别是 0.26、 0.06、 0.11mol。 最后得产品产率为 115.3%， 经 1 HNMR验证羧
甲基平均取代度 1.8， 羟丙基平均取代度 4.7， 产物简写为 CM2-HP5-P-CD。 实施例 11 与实施例 1基本相同， 但是其中 NaOH换为 KOH, 其 KOH、 氯乙酸乙酯、 1,2-环氧丙垸的投料量分别是 0.62、 0.24、 0.08mol o最后得钾盐的产物 （X = K、 Η) ， 产率为 119.9%， 经 1 HNMR验证羧甲基平均取代度 5.7， 羟丙基平均取代 度 3.2 , 产物简写为 CM6-HP3-P-CD。 实施例 12 与实施例 1基本相同， 但是其中 NaOH、 氯乙酸乙酯、 1，2-环氧丙烷的投料 量分别是 0.22、 0.04、 0.03mol。 最后得产品产率为 93.4%， 经 1 HNMR验证羧 甲基平均取代度 1.4， 羟丙基平均取代度 1.3， 产物简写为 CMt-HP!-P-CDa 实施例 13 与实施例 1基本相同， 但是其中 NaOH、 氯乙酸乙酯、 1，2-环氧丙垸的投料 量分别是 0.5、 0.18、 0.13mol。 最后得产品产率为 117.6%， 经 1 HNMR验证羧 甲基平均取代度 4.7， 羟丙基平均取代度 4.9， 产物简写为 CM5-HP5-p-CD。 实施例 14 与实施例 1基本相同， 但是其中 NaOH、 氯乙酸乙酯、 1，2-环氧丙烷的投料 量分别是 0.7、 0.24、 0.03mol。 最后得产品产率为 113.8%， 经 1 HNMR验证羧 甲基平均取代度 5.6， 羟丙基平均取代度 1.3， 产物简写为 CM6-HP^-CD。 实施例 15 与实施例 1基本相同， 但是其中 NaOH、 氯乙酸乙酯、 1,2-环氧丙烷的投料 量分别是 0.26、 0.06、 0.10mol。 最后得产品产率为 108.7%， 经 1 HNMR验证羧 甲基平均取代度 1.8， 羟丙基平均取代度 3.9， 产物简写为 CM2-HP4-P-CD。 实施例 16 与实施例 1基本相同， 但是其中 NaOH、 氯乙酸乙酯、 1，2-环氧丙垸的投料 量分别是 0.45、 0.15、 0.03mol。 最后得产品产率为 105.4%， 经 1 HNMR验证羧 甲基平均取代度 3.8， 羟丙基平均取代度 1.3， 产物简写为
It was basically the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.22, 0.04, and 0.12 mol, respectively. The yield of the product was 106.2%. The average degree of substitution of the carboxymethyl group was 1.3 by the 1 H NMR, and the average degree of substitution of the hydroxypropyl group was 4.6. The product was abbreviated as CMi-HPs-P-CDn. Example 9 was basically the same as Example 1, but Among them, NaOH was changed to KOH, and the amounts of KOH, ethyl chloroacetate and 1,2-propylene oxide were 0.33, 0.09, and 0.07 mol, respectively, and the products of the potassium salt (X = K, Η) were obtained. The yield was 110.9%, the average degree of substitution of carboxymethyl group was 2.9 by 1 H NMR, and the average degree of substitution of hydroxypropyl group was 2.8. The product was abbreviated as CM 3 -HP 3 -P-CD. Example 10 was substantially the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate and 1,2-propylene oxide were 0.26, 0.06, and 0.11 mol, respectively. The final product yield was 115.3%, and the carboxyl group was verified by 1 H NMR. The average degree of substitution of methyl groups was 1.8, and the average degree of substitution of hydroxypropyl groups was 4.7. The product was abbreviated as CM 2 -HP 5 -P-CD. Example 11 is basically the same as Example 1, except that NaOH is changed to KOH, and the amounts of KOH, ethyl chloroacetate, and 1,2-epoxypropionam are 0.62, 0.24, and 0.08 mol, respectively. The product (X = K, hydrazine), yield 119.9%, the average degree of substitution of carboxymethyl groups was 5.7 by 1 H NMR, and the average degree of substitution of hydroxypropyl groups was 3.2. The product was abbreviated as CM 6 -HP 3 -P-CD. Example 12 was substantially the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-propylene oxide were 0.22, 0.04, and 0.03 mol, respectively. The yield of the product was 93.4%. The average degree of substitution of the carboxymethyl group was 1.4 by the 1 H NMR, and the average degree of substitution of the hydroxypropyl group was 1.3. The product was abbreviated as CMT-HP!-P-CDa. Example 13 was basically the same as Example 1. However, the amounts of NaOH, ethyl chloroacetate and 1,2-epoxypropanone were 0.5, 0.18 and 0.13 mol, respectively. Finally, to obtain the product in a yield of 117.6 percent, verified by 1 HNMR average degree of substitution of carboxymethyl 4.7, average substitution degree 4.9 hydroxypropyl, the product is abbreviated as CM 5 -HP 5 -p-CD. Example 14 was basically the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate and 1,2-propylene oxide were 0.7, 0.24, and 0.03 mol, respectively. The final yield of the product was 113.8%. The average degree of substitution of the carboxymethyl group was 5.6 by 1 H NMR, and the average degree of substitution of the hydroxypropyl group was 1.3. The product was abbreviated as CM 6 -HP^-CD. Example 15 was substantially the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate and 1,2-propylene oxide were 0.26, 0.06, and 0.10 mol, respectively. The final yield of the product was 108.7%. The average degree of substitution of the carboxymethyl group was 1.8 by the 1 H NMR, and the average degree of substitution of the hydroxypropyl group was 3.9. The product was abbreviated as CM 2 -HP 4 -P-CD. Example 16 was substantially the same as Example 1, except that the amounts of NaOH, ethyl chloroacetate, and 1,2-epoxypropionam were 0.45, 0.15, and 0.03 mol, respectively. The yield of the final product was 105.4%. The average substitution degree of carboxymethyl group was 3.8 by 1 H NMR, and the average degree of substitution of hydroxypropyl group was 1.3. The product was abbreviated as
实施例 17 产品与青蒿素的包合及包合物的制备。 Example 17 Inclusion of a product with artemisinin and preparation of a clathrate.
以 ΗΡ-β-CD和 CM-P-CD为对照，本发明制备产物 CM3-HP6-P-CD在 pH6.86
溶液中的青蒿素包合常数 Ka (270nm)测定结果如下表: 环糊精 /青蒿素包合常数 Taking ΗΡ-β-CD and CM-P-CD as control, the preparation product CM 3 -HP 6 -P-CD of the present invention at pH 6.86 The artesian inclusion constant Ka (270 nm) in the solution is determined as follows: Cyclodextrin/artemisinin inclusion constant
CDD 拟 合 方 程 R2 KaCDD fitting equation R 2 Ka
ΗΡ-β-CD Y= = 0.0217χ + 18.99 0.9925 875.1ΗΡ-β-CD Y= = 0.0217χ + 18.99 0.9925 875.1
CM-p-CD Υ= 0.0102χ + 9.5804 0.9957 939.2CM-p-CD Υ = 0.0102χ + 9.5804 0.9957 939.2
CM3-HP6- -CD Υ- 0.0069Χ + 10.667 0.9934 1545.9 包合常数 Ka测定结果表明 CM3-HP6-P-CD对青蒿素的包合能力显著强于 现有产品 ΗΡ-β-CD和 CM-p-CD, CM-HP-p-CD二元取代环糊精衍生物有利于 对药物的包合， 相应的药物包合物更稳定且更容易制得。 精密称取 10克 CM3-HPH3-CD及 1.70g青蒿素， 与适量纯水混合充分研磨 3小时， 于 50°C减压干燥， 得白色固体包合物 （摩尔比 1 : 1 ) 。 取青蒿素、 CM3-HP6-P-CD、 青蒿素与 CM3-HP6-P-CD物理混合物、 CM 3 -HP 6 - -CD Υ- 0.0069Χ + 10.667 0.9934 1545.9 The inclusion constant Ka determination results show that the inclusion ability of CM 3 -HP 6 -P-CD for artemisinin is significantly stronger than the existing product ΗΡ-β- CD and CM-p-CD, CM-HP-p-CD binary substituted cyclodextrin derivatives facilitate drug inclusion, and the corresponding drug inclusion complexes are more stable and easier to manufacture. 10 g of CM 3 -HPH3-CD and 1.70 g of artemisinin were accurately weighed, mixed with an appropriate amount of pure water for 3 hours, and dried under reduced pressure at 50 ° C to obtain a white solid clathrate (molar ratio of 1:1). Take artemisinin, CM 3 -HP 6 -P-CD, artemisinin and CM 3 -HP 6 -P-CD physical mixture,
CM3-HPH3-CD/青蒿素包合物四样品各约 5.0mg， 进行差示扫描热分析： Α12Ο3 参比， 量程 ±50μν， 升温范围 40°C〜400°C， 升温速率 10°C/min， 得 DTA图 谱 （附图 9 ) 。 由产物、 混合物与包合物 DTA谱 （b、 c和 d) 比较可见： 混合 物 c 保留了青蒿素的熔点特征峰 （〜150°C ) ； 而包合物 DTA谱 d的青蒿素熔 点峰消失， 〜262.5 °C开始分解； 混合物 c于〜 259.0Ό即开始分解； 产物 b (CM3-HP6-p-CD样品） 则在〜 266.6'C才发生分解。 由此可判断 d是一新的物 相， 为 CM3-HP6-P-CD/青蒿素包合物。 溶解度测定： 分别取青蒿素、 青蒿素 /HP-p-CD包合物 （同法制备， 摩尔比 1： 1.16 ) 、 CM3-HP6-P-CD/青蒿素包合物 （摩尔比 1 : 1 ) 样品于 25°C测定溶解 度， 结果： 青蒿素 O.l lmg'ml-^ CM3-HP6-P_CD包合物 l.OSmg'ml- ΗΡ-β-CD 包合物 0.53mg'm , CM3-HP6-p-CD能够增加青蒿素溶解度 9.55倍，而 ΗΡ-β-CD 仅增加溶解度 4.81倍。 实施例 18 CM 3 -HPH3-CD/artemisinin inclusion complex four samples each about 5.0mg, for differential scanning thermal analysis: Α1 2 Ο 3 reference, range ± 50μν, temperature range 40 ° C ~ 400 ° C, heating rate At 10 ° C / min, the DTA map was obtained (Fig. 9). From the DTA spectra (b, c and d) of the product, mixture and inclusion complex, it can be seen that: mixture c retains the melting point characteristic peak of artemisinin (~150 ° C); and the melting point of artemisinin of the inclusion complex DTA spectrum d peak disappeared, ~262.5 ° C begins to decompose; c mixture began to decompose at ~ 259.0Ό; product of b (CM 3 -HP 6 -p- CD sample) was then decomposed at ~ 266.6'C. From this, it can be judged that d is a new phase, which is CM 3 -HP 6 -P-CD/artemisinin inclusion complex. Solubility determination: Artemisinin, artemisinin/HP-p-CD inclusion complex (prepared by the same method, molar ratio 1: 1.16), CM 3 -HP 6 -P-CD/artemisinin inclusion complex Molar ratio 1: 1 ) The solubility of the sample was measured at 25 ° C. Results: Artemisinin Ol lmg'ml-^ CM 3 -HP 6 -P_CD inclusion complex l.OSmg'ml- ΗΡ-β-CD inclusion complex 0.53 Mg'm, CM 3 -HP 6 -p-CD increased the solubility of artemisinin by 9.55 times, while ΗΡ-β-CD only increased the solubility by 4.81 times. Example 18
产物结构 NMR谱 Product structure NMR spectrum
HSQC、 COSY. ROESY、 I3CNMR和 1 HNMR谱 （溶剂 D2O， 图 1-5 ) ， 图 1-4的结构信号位移、 强弱、 峰形及其关联性质皆能对应符合于所标示的羧 甲基-羟丙基 - β -环糊精产物结构各单元特征， 证明本发明制备产物具有所设计 的结构。产物各取代基的结构 13CNMR (D2O) δ (ppm)谱归属： 177.594 ( C2，， -COOH )， 99.455 ( Ci，， -CH2-)， 76.278 ( Ci", O-CH2-) , 67.997 (C2"， -CHOH -)，
18.17 ( C3"， -CH3 ) ， 其环糊精主环基本保持原有碳谱特点。 iHNMR谱 （表） 显示： 由于取代基的影响， 产物结构中的环糊精葡糖环上质子位移相互交错呈 一组多重峰 （与 ΗΡ-β-CD谱类似） ， 而不象 β-CD那样清晰易于分辨， 葡糖环 的 -d-H质子也由 β-CD中的二重峰叠合为单峰； 羧甲基结构的亚甲基质子 0-CH2-COO-由于氧及羧基的共同影响向低场位移 （至 δ =5.28 ) ， 其位移及峰 形特点易与其他质子分辨而成为羧甲基及其取代度大小的重要判据； 羟丙基中 甲基质子基本保持了 ΗΡ-β-CD的二重峰特点，由于受其他质子影响较小且极易 分辨而成为羟丙基取代度的重要判据。 HSQC, COSY. ROESY, I3 CNMR and 1 H NMR spectra (solvent D 2 O, Figure 1-5), Figure 1-4 Structural signal displacement, strength, peak shape and their associated properties can correspond to the indicated The characteristics of each unit of the carboxymethyl-hydroxypropyl-β-cyclodextrin product structure demonstrate that the product of the present invention has a designed structure. Structure of each substituent of the product 13 CNMR (D 2 O) δ (ppm) Spectral assignment: 177.594 (C 2 ,, -COOH ), 99.455 ( Ci,, -CH 2 -), 76.278 ( Ci", O-CH 2 -) , 67.997 (C 2 ", -CHOH -), 18.17 ( C 3 ", -CH 3 ) , the cyclodextrin main ring basically retains the original carbon spectrum characteristics. The iHNMR spectrum (Table) shows: due to the influence of the substituent, the proton of the cyclodextrin glucose ring in the product structure The displacements are interlaced to form a set of multiple peaks (similar to the ΗΡ-β-CD spectrum), rather than as clear and easy to distinguish as β-CD, the -dH proton of the glucose ring is also superimposed by the doublet in β-CD. Single peak; the carboxymethyl structure of the methylene proton 0-CH 2 -COO- shifts to the low field due to the combined influence of oxygen and carboxyl groups (to δ = 5.28), and its displacement and peak shape are easily distinguished from other protons. An important criterion for the size of carboxymethyl groups and their degree of substitution; the methyl protons in hydroxypropyl groups basically retain the doublet characteristic of ΗΡ-β-CD, which is hydroxypropyl due to the small influence of other protons and easy to distinguish. An important criterion for the degree of substitution.
产物结构质子归属及结构单元 Product structure proton assignment and structural unit
ppm 质子 X归属 峰特征 结构单元 Ppm proton X attribution peak characteristics structural unit
1.13-1.15 3H:-CH3 d 羟丙基 1.13-1.15 3H:-CH 3 d Hydroxypropyl
3.40-4.30 3H:-O-CH2-CH-; m 羟丙基 3.40-4.30 3H:-O-CH 2 -CH-; m Hydroxypropyl
2H:-O-CH2- 葡糖环 2H:-O-CH 2 - glucose ring
6H:C2-H;C3-H;C4-H; 葡糖环 6H: C 2 -H; C 3 -H; C 4 -H; glucose ring
C5-H; 2 C6-H C 5 -H; 2 C 6 -H
5.07 s 葡糖环5.07 s glucose ring
5.28 2H:-0-CH2-COO- s 羧甲基 实施例 19 羧甲基-羟丙基 - β -环糊精的更昔洛韦包合物及稳定性试验 5.28 2H:-0-CH 2 -COO- s carboxymethyl Example 19 ganciclovir inclusion complex of carboxymethyl-hydroxypropyl-β-cyclodextrin and stability test
称取 CM2-HP3-P-CD样品 lO.Og与更昔洛韦原料药 l.Og混合， 加入 4ml水 研磨 0.5h ， 然后 45 Ό温度下减压干燥， 得 l lg粉末状产物。 DTA热分析： 产 物谱图中 235 °C：〜 260°C (更昔洛韦原料特征） 峰消失； 粉末与更昔洛韦原料稀 溶液 UV谱完全一致， 证明粉末产物为更昔洛韦包合物。 称取适量更昔洛韦包合物， 溶于葡萄糖注射液， 以 0.2μιη滤膜过滤， 配制 成更昔洛韦浓度 1.0mg_m 的包合物注射液 100ml , 市售更昔洛韦葡萄糖注射 液 （浓度 l.Omg'm ) 为对照， 取各注射液 20ml放置于 60Ό下， 于 0、 5、 10 天观察体系外观并测定更昔洛韦浓度， 计算注射液中更昔洛韦相对含量变化。 结果： 包合物注射液 5天及 10天体系无明显变化； 市售注射液颜色加深变黄。 HPLC ( C18柱，甲醇-水 -磷酸 3:97:0.01流动相，波长 284nm ) 含量 （％) 测定结 果见下表：
注射液 60Ό加速试验含量测定结果 The CM 2 -HP 3 -P-CD sample was weighed and mixed with the ganciclovir bulk drug 1.0 g, and added to 4 ml of water for 0.5 h, and then dried under reduced pressure at 45 Torr to obtain 1 g of a powdery product. DTA thermal analysis: 235 °C: ~ 260 °C in the product spectrum (the characteristics of ganciclovir) disappeared; the powder was completely consistent with the UV spectrum of the ganciclovir dilute solution, which proved that the powder product was ganciclovir Compound. Weigh the appropriate amount of ganciclovir inclusion compound, dissolve in glucose injection, filter with 0.2μιη filter, and prepare 100ml of inclusion compound injection of ganciclovir concentration 1.0mg_m. Commercially available ganciclovir glucose injection. (Concentration l.Omg'm) For the control, take 20ml of each injection and place it under 60Ό. Observe the appearance of the system at 0, 5, 10 days and determine the concentration of ganciclovir. Calculate the relative content of ganciclovir in the injection. . RESULTS: There was no significant change in the system of the inclusion compound for 5 days and 10 days. The color of the commercially available injections turned yellow. HPLC (C18 column, methanol-water-phosphoric acid 3:97:0.01 mobile phase, wavelength 284 nm) Content (%) The results are shown in the table below: Injection 60 Ό accelerated test content determination results
_ p 更昔洛韦相对含量 _ P relative content of ganciclovir
样 口口 Mouth
0 ^ 10天 0 ^ 10 days
市售注射液 100 97.8 94.7 Commercially available injection 100 97.8 94.7
包合物注射液 100 99.3 98.3 Inclusion Compound Injection 100 99.3 98.3
实施例 20 Example 20
蛋白质药物载体试验 Protein drug carrier test
本发明以 HP-p-CD、 CM-p-CD和 β-CD为对照， 进行了 CM-HP_p_CD对胰 岛素的包合能力、促进 Caco-2细胞体外胰岛素的转运作用、细胞毒性等体外实 验， 意外发现： CM-HP-p-CD与胰岛素有更强的包合和促进胰岛素跨膜转运的 效果， 而且对膜细胞基本无毒性。 此外， 通过动物实验， 发现 CM-HP-P-CD有 很好的促迸胰岛素的降糖能力。 （对照物 ΗΡ-β-CD不仅可用于化学药物的制 剂，还被认为是一种优良的蛋白大分子药物的载体，从而极大地开拓了 ΗΡ-β-CD 的应用领域。 ） 与胰岛素的包合作用强度： 羧甲基-羟丙基 环糊精与多肽类型药物-胰岛素的包合物制备及性能。本 发明的 CM2-HP6-p-CD以市售 HP-P-CD、 CM_p_CD和 β-CD为对照， 测定与胰 岛素的表观包合常数 Ka， 结果见下表： The invention adopts HP-p-CD, CM-p-CD and β-CD as control, and carries out in vitro experiments of CM-HP_p_CD on insulin incorporation, promotion of Caco-2 cell insulin in vitro, and cytotoxicity. Unexpectedly found: CM-HP-p-CD has a stronger inclusion of insulin and promotes transmembrane transport of insulin, and is essentially non-toxic to membrane cells. In addition, through animal experiments, it was found that CM-HP-P-CD has a good hypoglycemic ability to promote insulin. (The control ΗΡ-β-CD can be used not only as a preparation for chemical drugs, but also as a carrier for excellent protein macromolecular drugs, thus greatly expanding the application field of ΗΡ-β-CD.) Intensive strength: Preparation and properties of carboxymethyl-hydroxypropyl cyclodextrin and peptide type drug-insulin inclusion compound. The CM 2 -HP 6 -p-CD of the present invention is measured by the commercially available HP-P-CD, CM_p_CD and β-CD as the control, and the apparent inclusion constant Ka of insulin is determined. The results are shown in the following table:
多种环糊精的胰岛素表观包合常数 环糊精 方程 R2 a β-CD y = 0.0178x + 1.2572 0.9914 56.7Apparent inclusion constant of a variety of cyclodextrins. Cyclodextrin equation R 2 a β-CD y = 0.0178x + 1.2572 0.9914 56.7
ΗΡ-β-CD = 0.026x + 0.9614 0.9980 88.9ΗΡ-β-CD = 0.026x + 0.9614 0.9980 88.9
CM- -CD y = = 0.055x + 4.8394 0.9992 180.7CM- -CD y == 0.055x + 4.8394 0.9992 180.7
CM2-HP6- -CD y = 0.0149x + 2.6842 0.9939 31 1.2 CM 2 -HP 6 - -CD y = 0.0149x + 2.6842 0.9939 31 1.2
对比试验表明， CM2-HP6-P-CD包合胰岛素具有最大的表观包合常数 Ka， 揭示本发明制备的产物其包合性能明显优于对照产品 HP-p-CD、 CM-P-CD和
p-CD。 Comparative experiments showed that CM 2 -HP 6 -P-CD inclusion insulin has the largest apparent inclusion constant Ka, which reveals that the product prepared by the invention has better inclusion performance than the control products HP-p-CD, CM-P. -CD and p-CD.
胰岛素包合物制备： Preparation of insulin inclusion complex:
精密称取 6.0g本发明制备的 CM2-HP6-P-CD和 l.Og 胰岛素， 混合均匀后 加入 3mL水，研磨 lh，后 40°C下减压干燥得粉状包合物。溶解试验显示， lOmg 包合物能够完全溶解于 lmL水。 体外胰岛素的转运率测定： 6.0 g of CM 2 -HP 6 -P-CD and 1.0 g of insulin prepared by the present invention were accurately weighed, mixed uniformly, and then added with 3 mL of water, ground for 1 h, and then dried under reduced pressure at 40 ° C to obtain a powdery clathrate. The dissolution test showed that the lOmg inclusion complex was completely soluble in 1 mL of water. In vitro insulin transport rate determination:
样品： CM2-HP6-p-CD、 CM-P-CD, ΗΡ-β-CD, β-CD的胰岛素包合物 （环 糊精： 胰岛素包合比 =6: 1) 。 精密称取胰岛素和包合物适量， 将胰岛素和包合物分别溶于少量 0.01M的 盐酸中制成浓溶液， 以 pH7.2的 HBSS溶液稀释， 制备成浓度 15IUTI1L-1的样品溶 液， 15 IU'mL-1的胰岛素溶液为对照。 细胞接种于 12孔 Transwell (1.12cm2, 0.4μιπ) ， 板密度 2<105 个 'mL-1, 加 在顶端和基底端的培养基分别为 0.5mL及 1.5mL， 培养细胞约 3星期， 达到融合 分化。实验前用 HBSS洗去黏附在细胞表面的代谢物，取荧光素钠渗透量不超过 4.^g-hrl-cm-2, 同时 TEER值 500 Q'cm-2左右的 Transwell; 在膜上方的 Apical侧室 (简称 A侧)分别加入 15 Il^mL-1的胰岛素及包合物溶液， 在膜下方的 Basolateral 侧室 (简称 B侧)加入 1.5mLHBSS， 置于 37 °C转速为 50 rmin-1的空气摇床孵育， 分别于 30、 60、 90、 120、 150min从 BL池取样 0.5 ml， 并立即补加等量的 HBSS; 用 HPLC法测定样品中胰岛素含量。 每次实验重复 3次。 以胰岛素渗透量 （单位为 IU) 对时间 （t) 作图， 得胰岛素的渗透曲线。 渗 透系数 （^£) 的计算公式： Papp = (dQ/dt) / (A'C0) 其中 为单位时间内药物累计渗透量， ^为扩散面积 （^=1.12cm2) , Co 为 A侧药物初始浓度。 采用 t 检验， PO.05为有显著性差异。 结果见附图 10。 体外细胞模型胰岛素渗透转运试验中， 多种环糊精随时间的延长皆有胰岛素渗 透转运的促进作用， 其中 β-CD的促进作用最弱， 各种环糊精促进渗透的效果： CM2-HP6- -CD > CM- -CD> HP- -CD>p-CD, 其顺序与它们的胰岛素包合常数 完全一致。 本发明制备的 CM2-HIVP-CD具有最强的促进胰岛素渗透转运作用。 Samples: CM 2 -HP 6 -p-CD, CM-P-CD, ΗΡ-β-CD, β-CD insulin inclusion complex (cyclodextrin: insulin inclusion ratio = 6: 1). Accurately weigh the appropriate amount of insulin and the inclusion compound, dissolve the insulin and the inclusion compound in a small amount of 0.01M hydrochloric acid to prepare a concentrated solution, and dilute with a solution of pH 7.2 in HBSS to prepare a sample solution with a concentration of 15 IUTI1L- 1 , 15 The insulin solution of IU'mL- 1 was used as a control. The cells were seeded in 12-well Transwell (1.12 cm 2 , 0.4 μιπ), the plate density was 2<10 5 'mL- 1 , and the medium added to the apical and basal ends was 0.5 mL and 1.5 mL, respectively, and the cells were cultured for about 3 weeks. Fusion differentiation. Before the experiment, HBSS was used to wash away the metabolites adhering to the cell surface, and the permeation amount of sodium fluorescein did not exceed 4.^g-hr l -cm- 2 , and the TEER value was about 500 Q'cm- 2 . The Apical side chamber (referred to as the A side) was added with 15 Il^mL- 1 insulin and clathrate solution, and 1.5 mL HBSS was added to the Basolateral side chamber (B side) below the membrane, and the temperature was 50 rmin- 1 at 37 °C. Incubate on an air shaker, sample 0.5 ml from the BL pool at 30, 60, 90, 120, 150 min, and immediately add an equal amount of HBSS; determine the insulin content in the sample by HPLC. Repeat 3 times for each experiment. The penetration of insulin (in IU) versus time (t) gives the insulin penetration curve. The calculation formula of the permeability coefficient (^£): P app = (dQ/dt) / (A'C 0 ) where is the cumulative penetration of the drug per unit time, ^ is the diffusion area (^=1.12cm 2 ), and Co is A The initial concentration of the side drug. Using the t test, PO.05 was significantly different. The results are shown in Figure 10. In the in vitro cell model insulin permeation transport test, a variety of cyclodextrins promote insulin permeation transport over time, in which β-CD promotes the weakest effect, and various cyclodextrins promote permeation: CM 2 - HP 6 - -CD > CM- -CD> HP- -CD>p-CD, the order is exactly the same as their insulin inclusion constant. CM 2 -HIVP-CD prepared according to the invention has the strongest effect of enhancing insulin penetration and translocation.
研究结果显示：本发明制备的产物 CM-HP-P-CD对蛋白质 /多肽药物的包合 性能明显优于现有产品 ΗΡ-β-CD,因此在改进蛋白质 /多肽药物的口服给药方面 将具有很好的应用前景。 实施例 21
羧甲基-羟丙基 - β -环糊精与黄酮类型药物-葛根素的包合物制备及溶血试 验。 精密称取 10g本发明的 CM3-HP3-P-CD与 2.8g葛根素混合均匀（摩尔比 1 : 1 ) ， 于 60Ό温度下加热即熔融， 保持温度 lh， 然后于 25'C下冷却， 24小时 后研磨， 得粉末状产物； DTA谱证明产物为葛根素的 CM3-HIVP-CD包合物。 证明本发明产物对多羟基类的黄酮药物容易制得其包合物， 制备过程中无须添 加任何溶剂助剂， 特别适合于敏感性药物包合物的制备。 另取 lO.Og本发明的 CM2-HPH3-CD和 2.0g葛根素， 同法制得葛根素： CM2-HP5-p-CD包合物（质量比 5: 1 )，取包合物 6g溶于 NaCl注射液， 以 0.2μπι 滤膜过滤， 配制成葛根素浓度 Z.Omg.ml-1的注射液 500ml， 溶血试验 （文献方 法： 《药物研究技术指导原则（2005 )》，中国医药科技出版社，2006， P116-132 ) 观测不到明显溶血作用 （溶血率 <5% ) ， 相同浓度对比的普通市售葛根素注射 液 （葛根素浓度 S.Omg'ml-1 ) 则存在明显溶血作用 （溶血率 09%) 。 试验证明， 使用本发明制备的 CM-HP-P-CD能够有效降低药物溶血作用， 从而增强临床用药的安全性。 The results of the study show that the inclusion of the product CM-HP-P-CD of the present invention on the protein/polypeptide drug is superior to the existing product ΗΡ-β-CD, and thus will improve the oral administration of the protein/polypeptide drug. Has a good application prospects. Example 21 Preparation of carboxymethyl-hydroxypropyl-β-cyclodextrin and flavonoid drug-puerarin and hemolysis test. Weigh accurately 10g of the CM 3 -HP 3 -P-CD of the present invention is uniformly mixed with 2.8g of puerarin (molar ratio of 1:1), melted at 60 ° C, heated at a temperature of 1 h, and then cooled at 25 ° C. After 24 hours, it was ground to obtain a powdery product; the DTA spectrum confirmed that the product was a CM 3 -HIVP-CD clathrate of puerarin. It is proved that the product of the present invention can easily prepare a clathrate compound for the polyphenolic flavonoid drug, and it is not necessary to add any solvent auxiliary agent in the preparation process, and is particularly suitable for the preparation of the sensitive drug clathrate. Another lO.Og CM 2 -HPH3-CD of the present invention and 2.0 g of puerarin, the same method to prepare puerarin: CM 2 -HP 5 -p-CD inclusion complex (mass ratio 5: 1), the inclusion complex 6g dissolved in NaCl injection, filtered with 0.2μπι filter, prepared into 500ml of injection of puerarin concentration Z.Omg.ml- 1 , hemolysis test (literature method: "Guidelines for Drug Research Technology (2005)", Chinese medicine Science and Technology Press, 2006, P116-132) No obvious hemolysis (hemolytic rate <5%) was observed, and the common commercial puerarin injection (puerarin concentration S.Omg'ml- 1 ) with the same concentration was observed. Hemolysis (hemolytic rate 09%). Tests have shown that the use of the CM-HP-P-CD prepared by the present invention can effectively reduce the hemolysis of the drug, thereby enhancing the safety of the clinical drug.
实施例 22 Example 22
产品 IR谱 Product IR spectrum
本发明制备的不同取代度羧甲基-羟丙基 - P -环糊精系列产物 KBr压片测 定 IR光谱， 与 β-CD相比， CM-HP-p-CD产物在 2969cm-1和 1461cm-1有明显吸 收 （羟丙基上甲基的反对称伸缩振动和对称弯曲振动吸收） ， 1420cm-1和 1617cm-1峰证明存在羧甲基取代基。 The IR spectrum of KBr pellets of different substituted carboxymethyl-hydroxypropyl-P-cyclodextrin products prepared by the present invention is compared with β-CD, and the CM-HP-p-CD products are 2969cm- 1 and 1461cm. - 1 has significant absorption (antisymmetric stretching vibration of hydroxypropyl group and symmetric bending vibration absorption), and 1420 cm- 1 and 1617 cm- 1 peaks indicate the presence of a carboxymethyl substituent.
实施例 23 Example 23
产品热分析 Product thermal analysis
本发明制备的不同取代度羧甲基-羟丙基 环糊精系列产物皆为白色无 定型粉末状固体， 易吸潮， 充分干燥后产品仍含 4%〜11%不等的包合水。 所有 产物加热分解而无固定熔点，分解点随取代度的增加（分子量递增）在 230X〜 280°C范围逐步升高。 以 CM2-HP6-p-CD的 DTA图谱（图 6 ) 为例， 其样品（e) 特征： 90°C以下有着比 p-CD、 CM-p-CD和 ΗΡ-β-CD更宽的脱水吸热峰； 100 °C〜175 °C体系比热缓慢增大， 175 ° ( 〜 270°C比热又渐变小； 270°C开始分解； 300〜335°C呈多重放热峰。 多样品的 DTA图谱比较可见， 各温度区间的
CM2-HP6- -CD热性质皆与其他对比样品存在明显 The products of different degrees of substitution of carboxymethyl-hydroxypropyl cyclodextrin prepared by the invention are all white amorphous powdery solids, which are easy to absorb moisture, and the products still contain 4%~11% of inclusion water after sufficient drying. All products are thermally decomposed without a fixed melting point, and the decomposition point is gradually increased in the range of 230X to 280 °C as the degree of substitution (increase in molecular weight) increases. Taking the DTA map of CM 2 -HP 6 -p-CD (Fig. 6 ) as an example, the sample (e) features: 90 ° C or less is wider than p-CD, CM-p-CD and ΗΡ-β-CD. Dehydration endothermic peak; 100 °C ~ 175 °C system slowly increases in specific heat, 175 ° (~ 270 ° C is slightly smaller than heat; 270 ° C begins to decompose; 300 ~ 335 ° C shows multiple replay heat peaks. The DTA spectra of multiple samples are visible, and the temperature ranges are CM 2 -HP 6 - -CD thermal properties are significantly present with other comparative sample
实施例 24 Example 24
产品取代度 Product substitution
产物 ¾HNMR特征为测定产品取代度控制质量提供了方便， 以 δ =5.07峰 (环糊精葡糖环 1位 Η)面积积分标记为 7 (每摩尔 β -环糊精含 7个葡糖环）， 则测定 δ =5.28的单峰（-0-CH2-)面积的 1/2值即为 CM基取代度，其 δ =1.13〜 1.15二重峰 （-CHOH-CH3) 面积值 1/3即为 HP基取代度 （图 5 ) 。 采用相同 的方法， 由 iHNMR得到的本发明制备的部分产物取代度见表 2: The 3⁄4 H NMR characteristics of the product provide convenience for determining the degree of product substitution control, with an area integral of δ = 5.07 peak (cyclodextrin glucose ring 1 position 标记) as 7 (containing 7 glucose rings per mole of β-cyclodextrin) ), the 1/2 value of the single peak (-0-CH2-) area of δ = 5.28 is determined as the CM group substitution degree, and the δ = 1.13 to 1.15 doublet (-CHOH-CH 3 ) area value is 1/ 3 is the HP base substitution degree (Figure 5). Using the same method, the partial product substitution degree prepared by the present invention obtained by iH NMR is shown in Table 2:
10种不同羧甲基-羟丙基 - β -环糊精的平均取代度 实际平均取代度 总取代度 平均分子量 Average degree of substitution of 10 different carboxymethyl-hydroxypropyl-β-cyclodextrins Actual average degree of substitution Total degree of substitution Average molecular weight
产物 Product
n m z (g-mol"1 ) Nmz (g-mol" 1 )
CMi-HP2-p-CD 1.2 1.8 3 1309 CMi-HP 2 -p-CD 1.2 1.8 3 1309
CMi-HPs- -CD 1.1 3.3 4 1367 CMi-HPs- -CD 1.1 3.3 4 1367
CM2-HP2- -CD 1.9 2.2 4 1367 CM 2 -HP 2 - -CD 1.9 2.2 4 1367
CMi-HP4- -CD 1.2 4.1 5 1425 CMi-HP 4 - -CD 1.2 4.1 5 1425
CM3-HP2-P-CD 3.3 2.4 5 1425 CM 3 -HP 2 -P-CD 3.3 2.4 5 1425
CMi-HPs- -CD 1.3 4.6 6 1483 CMi-HPs- -CD 1.3 4.6 6 1483
CM3-HP3- -CD 2.9 2.8 6 1483 CM 3 -HP 3 - -CD 2.9 2.8 6 1483
CM2-HP5-P-CD 1.8 4.7 7 1541 CM 2 -HP 5 -P-CD 1.8 4.7 7 1541
CM2-HP6-p-CD 2.1 5.8 8 1599 CM 2 -HP 6 -p-CD 2.1 5.8 8 1599
CM3-HP6-p-CD 2.7 5.9 9 1657 CM 3 -HP 6 -p-CD 2.7 5.9 9 1657
CM6-HP3-p-CD 5.7 3.2 9 1657 CM 6 -HP 3 -p-CD 5.7 3.2 9 1657
实施例 25 Example 25
理化性质试验 Physical and chemical properties test
本发明制备的羧甲基-羟丙基- 3 -环糊精具有 ΗΡ-β-CD相似的溶解特性，系 列产物皆极易溶于水、 稀酸 /稀碱液， 也极易溶于甲醇 /乙醇水溶液， 易溶于纯 甲醇 /纯乙醇， 在常用有机溶剂如正己烷、 乙酸乙酯、 三氯甲烷、 丙酮等中难溶 或不溶解， 不同取代度产物溶解性能基本无差异。 羧甲基-羟丙基 - e -环糊精在 纯乙醇中的溶解度可达羟丙基-磺丁基 环糊精的 2.5倍以上， 良好的溶解特 性将十分有利于高脂溶性药物包合物的制备， 对改进难溶药物性能研制新型制
剂产品具有十分重要的应用价值。 以 CM3-HP3-p-CD为例的溶解性试验结果见 下表. · The carboxymethyl-hydroxypropyl-3-cyclodextrin prepared by the invention has the similar solubility characteristics of ΗΡ-β-CD, and the series products are highly soluble in water, dilute acid/dilute lye, and also very soluble in methanol. / Ethanol aqueous solution, soluble in pure methanol / pure ethanol, difficult to dissolve or insoluble in common organic solvents such as n-hexane, ethyl acetate, chloroform, acetone, etc., the solubility properties of the products with different degrees of substitution are basically no difference. The solubility of carboxymethyl-hydroxypropyl-e-cyclodextrin in pure ethanol can be more than 2.5 times that of hydroxypropyl-sulfobutylcyclodextrin. Good solubility characteristics will be very beneficial for high fat-soluble drug inclusion complexes. Preparation, research and development of new systems for improving the performance of poorly soluble drugs Agent products have important application value. The solubility test results using CM 3 -HP 3 -p-CD as an example are shown in the table below.
CM3-HP3-P-CD溶解性试验 * CM 3 -HP 3 -P-CD Solubility Test*
样品量 溶剂加入量 Sample amount
溶 剂 溶质 /溶剂 溶解情况 结论 Solvent Solute / Solvent Dissolution Conclusion
(g) (ml) (g) (ml)
水 0.50 0.50 1/1 全部溶解 极易溶解 Water 0.50 0.50 1/1 All dissolved Very soluble
O.lmol-L"1 HC1 0.50 0.50 1/1 全部溶解 极易溶解O.lmol-L" 1 HC1 0.50 0.50 1/1 All dissolved very soluble
O.lmol-L 'NaOH 0.50 0.50 1/1 全部溶解 极易溶解 甲 醇 0.10 0.60 1/ 6 全部溶解 易溶 乙 醇 0.10 0.90 1/ 9 全部溶解 易溶 乙 腈 0.10 2.70 1/ 27 全部溶解 溶解 正己烷 0.01 100 1/10000 溶液混浊 不溶 乙酸乙酯 0.01 100 1/10000 溶液混浊 不溶 三氯甲烷 0.01 100 1/10000 溶液混浊 不溶 丙酮 0.01 100 1/10000 溶液混浊 不溶 O.lmol-L 'NaOH 0.50 0.50 1/1 All dissolved very soluble methanol 0.10 0.60 1/ 6 All dissolved easily soluble ethanol 0.10 0.90 1/ 9 All dissolved readily soluble acetonitrile 0.10 2.70 1/ 27 All dissolved dissolved n-hexane 0.01 100 1/10000 solution turbid insoluble ethyl acetate 0.01 100 1/10000 solution turbid insoluble chloroform 0.01 100 1/10000 solution turbid insoluble acetone 0.01 100 1/10000 solution turbid insoluble
* 溶解度： 《中华人民共和国药典》 2000年版二部凡例溶解度项下测试。 本发明制备的羧甲基-羟丙基 - -环糊精系列产物皆具有光学活性， 比旋 度 [ α ]随取代度的变化在 +85°〜+107°间波动， 但比旋度值明显低于单一取代基 的羟丙基 -β-环糊精 （+ 128°〜+145° ) 、 羧甲基 -β-环糊精 （+ 174.0°〜+180° ) 和 母体 β-环糊精（ + 162°) 的比旋度， 也明显低于这些已知环糊精产品的混合物的 比旋度， 与羟丙基-磺丁基-环糊精比旋度 （+99.0°〜+171° ) 亦有明显差异。 部 分产物 [ α ]的测定值见下表： 部分产物的比旋光度 目标化合物 比旋光度 [α* Solubility: "People's Republic of China Pharmacopoeia" 2000 version of the second part of the solubility test. The carboxymethyl-hydroxypropyl-cyclodextrin series products prepared by the invention all have optical activity, and the specific rotation [α] fluctuates with the degree of substitution between +85° and +107°, but the specific rotation value Hydroxypropyl-β-cyclodextrin (+128°~+145°), carboxymethyl-β-cyclodextrin (+ 174.0°~+180°) and parent β-cyclodide significantly lower than a single substituent The specific rotation of the fine (+ 162°) is also significantly lower than the specific rotation of the mixture of these known cyclodextrin products, and the specific rotation of hydroxypropyl-sulfobutyl-cyclodextrin (+99.0°~+ 171°) There are also significant differences. The measured values of some products [α ] are shown in the following table: Specific optical rotation of some products Target compound Specific optical rotation [α
CM3-HP2-P-CD +90.43 CM 3 -HP 2 -P-CD +90.43
CM3-HP3- -CD + 106.77 CM 3 -HP 3 - -CD + 106.77
CM2-HP6- -CD +87.88 CM 2 -HP 6 - -CD +87.88
CM3-HP6-p-CD +85.98 CM 3 -HP 6 -p-CD +85.98
CM6-HP3-P-CD +102.39
比旋度测试说明： 本发明制备的羧甲基-羟丙基 - β -环糊精是羧甲基和羟丙 基混合取代的环糊精衍生物而不是羟丙基- β -环糊精与羧甲基- β -环糊精二者 的简单混合物。 CM 6 -HP 3 -P-CD +102.39 Specific rotation test description: The carboxymethyl-hydroxypropyl-β-cyclodextrin prepared by the present invention is a carboxymethyl and hydroxypropyl mixed-substituted cyclodextrin derivative instead of hydroxypropyl-β-cyclodextrin. A simple mixture with both carboxymethyl-β-cyclodextrin.
在 HPLC测定 ΗΡ-β-CD和 β-CD的一般色谱条件下， 本发明制备的羧甲基- 羟丙基- β -环糊精与 HP-p-CD、 β-CD和 CM-p-CD存在明显不同的 HPLC色谱 特征， 易于识别本发明产物。 The carboxymethyl-hydroxypropyl-β-cyclodextrin prepared by the present invention and HP-p-CD, β-CD and CM-p- under the general chromatographic conditions for the determination of ΗΡ-β-CD and β-CD by HPLC. The CD has significantly different HPLC chromatographic characteristics, making it easy to identify the products of the invention.
色谱条件 色谱柱： 氨基柱 (4.6mmx250mm,5 m); 折光检测器； 流动相：乙 腈： 水 (67: 33)； 流速： l.Oml/min; 柱温 30°C。 精密称取 P_CD、 CM- -CD. ΗΡ-β-CD, 以及这三者的物理混合物 （质量比 1 : 2： 6 ) 、 合成的羧甲基 -羟丙 基 -β-环糊精（以 CM2-HP6-p-CD为例）各 O.lg，分别用流动相溶解并定容至 5ml， 进样 20μ1测定， 得色谱图： β-CD为色谱单峰 （11.3min) ， ΗΡ-β-CD也近似 为色谱单峰 （6.1〜6.4min ) ， CM^-CD先为宽平色谱峰 （14.2min) 再现多重 色谱峰（21.0〜35.0min) , CM2-HP6-P-CD (附图 7)则先出现多重色谱峰 ( 7.3 - 11.2min ) 后再现宽色谱峰 （17〜35min ) ， CM2-HP6-P_CD色谱中无 ΗΡ-β-CD 的 6.1〜6.4min特征单峰 （显示无 ΗΡ-β-CD存在） ， 也基本无 11.3min的 β-CD 特征单峰 （β-CD残留少） 。 p-CD、 CM-P-CD和 ΗΡ-β-CD的三者物理混合物色 谱（附图 8 ) 显示 β-CD和 ΗΡ-β-CD高的特征单峰， 其后出现 CM-p-CD低矮的 宽色谱峰， 色谱峰形及出峰时间均与本发明产物具有明显差异。 Chromatographic conditions column: Amino column (4.6mm x 250mm, 5 m); Refractive detector; Mobile phase: Acetonitrile: Water (67: 33); Flow rate: l.Oml/min; Column temperature 30 °C. Accurately weigh P_CD, CM--CD. ΗΡ-β-CD, and the physical mixture of these three (mass ratio 1: 2: 6), synthetic carboxymethyl-hydroxypropyl-β-cyclodextrin CM 2 -HP 6 -p-CD as an example) each O.lg, were dissolved in mobile phase and volume to 5ml, 20μ1 sample measured chromatogram obtained: β-CD as a single chromatographic peak (11.3min), ΗΡ -β-CD is also approximated as a single peak of chromatography (6.1~6.4min), and CM^-CD firstly reproduces multiple chromatographic peaks (21.0~35.0min) for broad-spectrum peaks (14.2min), CM 2 -HP 6 -P- The CD (Fig. 7) showed multiple peaks (7.3 - 11.2min) and reproduced the broad peak (17~35min). The CM 2 -HP 6 -P_CD chromatogram showed no 6.1-β-CD 6.1~6.4min. A single peak (showing the presence of no sputum-β-CD) also had essentially no single peak of β-CD characteristic of 11.3 min (less β-CD residue). The physical mixture chromatogram of p-CD, CM-P-CD and ΗΡ-β-CD (Fig. 8) shows a characteristic single peak of β-CD and ΗΡ-β-CD, followed by CM-p-CD The low broad peaks, peak shape and peak time are all significantly different from the products of the present invention.
色谱特征及比旋度测定说明： 本发明产物羧甲基-羟丙基 - β -环糊精是羟丙 基和羧甲基混合取代的产物，而不是羟丙基 环糊精与羧甲基 - β -环糊精的简 单物理混合物。 Chromatographic characteristics and specific rotation determination: The product of the invention, carboxymethyl-hydroxypropyl-β-cyclodextrin, is a product of mixed substitution of hydroxypropyl and carboxymethyl, instead of hydroxypropyl cyclodextrin and carboxymethyl - A simple physical mixture of β-cyclodextrin.
实施例 26 Example 26
产物纯度测定 Product purity determination
本发明的羧甲基-羟丙基 - 3 -环糊精由 β-环糊精与氯乙酸乙酯和 1,2-环氧丙 垸在水溶液中强碱催化反应而得，体系杂质为残留的 β-环糊精和反应试剂的碱 催化水解产物： 羟基乙酸和 1,2-丙二醇， 采用调整 ρΗ的透析纯化技术， 可极 大降低 β-环糊精残留量和 1,2-丙二醇含量，羟基乙酸分子量小并以负离子形式 存在而更易于除净， 从而使本发明的羧甲基-羟丙基 环糊精产品具有高的纯 度， 并具备良好的产品溶解和包合性能， 显著低的溶血性和更好的安全性。 本 发明的羧甲基-羟丙基 - β -环糊精产品中需控制的主要杂质及限量如下表：
羧甲基-羟丙基 环糊精中的主要杂质 杂 质 杂质来源 杂质限量 测定方法 环糊精 残 留 <1.5% HPLC The carboxymethyl-hydroxypropyl-3-cyclodextrin of the invention is obtained by reacting β-cyclodextrin with ethyl chloroacetate and 1,2-epoxypropanone in a strong base in an aqueous solution, and the system impurities are residual Base-catalyzed hydrolysates of β-cyclodextrin and reagents: glycolic acid and 1,2-propanediol, using dialysis purification technology to adjust ρΗ, can greatly reduce the residual amount of β-cyclodextrin and 1,2-propanediol The glycolic acid has a small molecular weight and exists in the form of an anion to be more easily removed, so that the carboxymethyl-hydroxypropyl cyclodextrin product of the invention has high purity and good product dissolution and inclusion properties, and is significantly low. Hemolysis and better safety. The main impurities and limits to be controlled in the carboxymethyl-hydroxypropyl-β-cyclodextrin product of the present invention are as follows: Main Impurities in Carboxymethyl-Hydroxypropyl Cyclodextrin Impurity Source Impurity Limit Determination Method Cyclodextrin Residue <1.5% HPLC
1,2-丙二醇 水解产物 <2.5% GC 1 HPLC 产品纯化处理后仅含有的微量羟基乙酸 （约 10〜20ppm ) ， 该杂质与目前 生物医学工程领域的新型材料乳酸一羟基乙酸共聚物 (PLGA) 的降解产物完全 相同。 广泛研究已经证明， PLGA有着良好的生物相容性和生物降解性能， 安 全性好， 可制作为人工导管、 药物缓释载体、 组织工程支架材料， 目前已获得 快速发展。 1,2-propanediol hydrolysate <2.5% GC 1 HPLC product contains only traces of glycolic acid (about 10~20ppm) after purification. This impurity is related to the new material of biomedical engineering, lactic acid monoglycolic acid copolymer (PLGA). The degradation products are identical. Extensive research has shown that PLGA has good biocompatibility and biodegradability, and it has good safety. It can be made into artificial catheter, drug delivery carrier, tissue engineering scaffold material, and has been rapidly developed.
实施例 27 Example 27
药物包合试验 Drug inclusion test
采用紫外分光光度法测定系列 CM-HP-P-CD与葛根素、 更昔洛韦、 盐酸 特比萘芬、 苯甲酸钠等富含羟基的黄酮、 N杂环、 阳离子及阴离子结构等典型 结构药物的包合常数 Ka (王亚娜，陆亚鹏，任勇等.环糊精及衍生物 /药物包合常数 的测定方法及其应用 [J].药学进展, 2004，28(1):23. ) ， 以检验本发明产品的包合 作用及性能， 以 CM3-HP3-P-CD为例， 考察比较其与其它环糊精衍生物的包合 性能， 结果见下表： Determination of CM-HP-P-CD and puerarin, ganciclovir, terbinafine hydrochloride, sodium benzoate and other hydroxy-containing flavonoids, N-heterocyclic, cationic and anionic structures by ultraviolet spectrophotometry Inclusion constant Ka (Wang Yana, Lu Yapeng, Ren Yong et al. Determination of cyclodextrin and derivatives/drug inclusion constants and their applications [J]. Progress in Pharmaceutical Sciences, 2004, 28(1): 23.), The inclusion and performance of the product of the present invention were examined. Taking CM 3 -HP 3 -P-CD as an example, the inclusion properties of the product with other cyclodextrin derivatives were examined and compared. The results are shown in the following table:
产品与典型结构药物的包合常数 结构 波长 包合常数 Ka Inclusion constant of product and typical structural drug Structure Wavelength Inclusion constant Ka
ΗΡ-β-CD CM-p-CD CM3-HP3- -CD 葛根素 黄 酮 250 519.8 702.7 1746.7 更昔洛韦 N杂环 252 650.2 435.5 737.6 盐酸特比萘芬 阳离子 282 897.7 771.3 964.1 苯甲酸钠 阴离子 224 784.2 1 134.9 3438.3 结果显示， 本发明的羧甲基-羟丙基- β -环糊精与多种结构类型药物皆有较 强的包合作用， 由于本发明的羧甲基-羟丙基 - β -环糊精易形成氢键， 因此， 对 含羧基及羟基结构的药物显示极高的包合能力。 进一步测定本发明的不同取代 度产物与上述药物的包合常数， 发现随取代度 （Ζ) 改变， 其包合常数约在上
述 Ka值 ±5%〜士 9% 范围内波动， Z = 4〜7范围产物有较大的 Ka值， 在更大 的取代度范围内 （Z = 2〜9) 即使 Ka有所降低的情况下， 不同取代度产物包合 上述药物的 Ka值仍然大于上述环糊精对照产品。 配制 CM3-HP3-P-CD及对照 环糊精衍生物产品的 3%溶液， 加入过量上述药物， 搅拌 10小时， 过滤测定药 物溶解度结果见下表- 环糊精溶液中的药物溶解度 溶解度 mg-ml"1 ΗΡ-β-CD CM-p-CD CM 3 -HP 3 - -CD puerarin flavonoid 250 519.8 702.7 1746.7 ganciclovir N heterocyclic 252 650.2 435.5 737.6 terbinafine hydrochloride 282 897.7 771.3 964.1 sodium benzoate anion 224 784.2 1 134.9 3438.3 The results show that the carboxymethyl-hydroxypropyl-β-cyclodextrin of the present invention has strong intercalation with various structural types of drugs due to the carboxymethyl-hydroxypropyl group of the present invention. The β-cyclodextrin is liable to form a hydrogen bond, and therefore exhibits an extremely high inclusion ability for a drug having a carboxyl group and a hydroxyl group structure. Further determining the inclusion constants of the different degree of substitution products of the present invention and the above drugs, and found that the inclusion constant is about the same as the degree of substitution (Ζ). The Ka value ranges from ±5% to 9% within the range of fluctuations, and the Z = 4 to 7 range product has a larger Ka value, in the range of greater substitution (Z = 2 to 9) even if the Ka is reduced Next, the Ka value of the above-mentioned drugs containing different substitution products is still greater than the above cyclodextrin control product. Prepare a 3% solution of CM 3 -HP 3 -P-CD and control cyclodextrin derivative product, add the above drug, stir for 10 hours, and determine the solubility of the drug by filtration. See the following table - Solubility Solubility in Cyclodextrin Solution Mg-ml" 1
ΗΡ-β-CD CM- -CD CM2-HP3-P-CD 葛根素 4.62 13.77 12.13 16.68 更昔洛韦 4.27 36.22 37.41 49.53 特比萘芬 1.12 5.67 5.32 7.86 苯甲酸 3.4 4.26 4.48 5.33 ΗΡ-β-CD CM- -CD CM 2 -HP 3 -P-CD Puerarin 4.62 13.77 12.13 16.68 Ganciclovir 4.27 36.22 37.41 49.53 Terbinafine 1.12 5.67 5.32 7.86 Benzoic acid 3.4 4.26 4.48 5.33
Ka测定及增溶试验表明本发明制备产物具有优良的包合能力。 实施例 28 Ka determination and solubilization tests show that the products prepared by the present invention have excellent inclusion ability. Example 28
溶血试验（文献方法： 《药物研究技术指导原则 （2005) 》 ， 中国医药科技 出版社， 2006， P116-132) Hemolysis test (literature method: Guiding Principles of Drug Research (2005), China Medical Science and Technology Press, 2006, P116-132)
以 ΗΡ-β-CD和 CM-P-CD为对照， 选取本发明的 11种化合物溶血试验结果 见表 13。 结果表明 :1) 在测试浓度下， 本发明制备的化合物溶血性均明显小于 对照； 2) ll.Omg.m l以下浓度， 本发明制备的化合物几乎观察不到溶血现象， ll.O lSJmg'ml-1产生轻度溶血（溶血率 5%);3)化合物浓度增大至 SS.Omg'ml-1 以上产生明显溶血 （溶血率 50%) ， 但所需浓度显著高于对照； 4) 取代度 Z = 4〜7的化合物溶血性较小。
各受试样品溶血实验结果 Taking ΗΡ-β-CD and CM-P-CD as controls, the results of the hemolysis test of 11 compounds of the present invention were selected as shown in Table 13. The results showed that: 1) at the test concentration, the hemolytic property of the compound prepared by the invention was significantly smaller than that of the control; 2) the concentration of ll.Omg.ml or less, the compound prepared by the invention hardly observed hemolysis, ll.O lSJmg'ml - 1 produces mild hemolysis (hemolytic rate 5%); 3) compound concentration increased to SS.Omg 'ml- 1 or more to produce significant hemolysis (hemolytic rate 50%), but the required concentration is significantly higher than the control; 4) replaced Compounds with degree Z = 4 to 7 have less hemolytic activity. Hemolysis test results of each test sample
溶血 5% 时浓度溶血 50% 时浓度 Hemolysis 5% concentration concentration hemolysis 50% concentration
( mg-ml"1 ) (mg-ml"1 )
( mg-ml" 1 ) (mg-ml" 1 )
CM2-HP2-P-CD 11.8 42.3 CM 2 -HP 2 -P-CD 11.8 42.3
CM1-HP4-p-CD 12.4 37.2 CM 1 -HP 4 -p-CD 12.4 37.2
CM3-HP2-p-CD 13.3 47.9CM 3 -HP 2 -p-CD 13.3 47.9
CM3-HP3-p-CD 12.8 39.8 CM 3 -HP 3 -p-CD 12.8 39.8
CM2-HP5-p-CD 12.6 41.4 CM 2 -HP 5 -p-CD 12.6 41.4
CM2-HP6- -CD 11.9 35.2 CM 2 -HP 6 - -CD 11.9 35.2
CM3-HP6- -CD 11.2 33.6 CM 3 -HP 6 - -CD 11.2 33.6
CM6-HP3-p-CD 11.6 35.1 CM 6 -HP 3 -p-CD 11.6 35.1
HP- β -CD 7.7 17.7 HP-β-CD 7.7 17.7
CM- β -CD 2.2 18.8 葛根素常规注射液 （使用浓度 0.4mg.ml -1〜 O.Smg.ml-1 ) 临床时常发生与溶 血有关的不良反应。 以 CM2-HP5-P-CD制备葛根素： CM2-HP5-P-CD包合物 （质 量比 1 :5 )，配制成葛根素浓度 S.Omg.ml-1的注射液， 观测不到明显溶血作用（溶 血率 <5% ) ， 相同浓度的葛根素对比注射液 （葛根素浓度 Z.Omg'm ) 则存在 明显溶血作用 （溶血率>9%) 。 试验表明， 本发明制备的 CM-HP-p-CD不仅自 身溶血作用小， 还能够有效降低被包合药物的溶血作用， 提高用药安全性。 CM-β-CD 2.2 18.8 Puerarin conventional injection (using concentration 0.4mg.ml - 1 ~ O.Smg.ml- 1 ) Clinically, adverse reactions related to hemolysis often occur. Preparation of puerarin by CM 2 -HP 5 -P-CD: CM 2 -HP 5 -P-CD inclusion complex (mass ratio 1:5), formulated into an injection of puerarin concentration S.Omg.ml- 1 , No obvious hemolysis was observed (hemolysis rate <5%), and the same concentration of puerarin contrast injection (puerarin concentration Z.Omg'm) showed significant hemolysis (hemolysis rate >9%). Tests have shown that the CM-HP-p-CD prepared by the invention not only has a small self-hemolytic effect, but also can effectively reduce the hemolysis of the encapsulated drug and improve the safety of administration.
实施例 29 Example 29
产物急性毒性试验及动物观察 Acute toxicity test and animal observation
以文献方法： （ 《药物研究技术指导原则 （2005 ) 》 ， 中国医药科技出版 社， 2006， P83-93 ) 选取多个不同取代度的本发明的 CM-HP-p-CD样品， 考察 产物的急性毒性， 结果显示小鼠灌胃给药， 样品剂量达 SOOO mg'kg-1时， 未见 受试动物死亡， 也未观察到明显的毒副反应。 以静脉注射给药剂量达 2000 mg^g-1时， 未见受试动物死亡， 也未观察到明显的毒副反应。 本试验未能测出 选取样品的两种给药途经的 LD5«), 其最大耐受量分别为 8000 mg-kg-1和 2000 mg-kg-1 , 结果提示， 本发明制备的 CM-HP-P-CD毒性很低。
CM-HP-p-CD急性毒性
By literature method: (Guidelines for Technical Guidance in Drug Research (2005), China Medical Science and Technology Press, 2006, P83-93) A plurality of samples of CM-HP-p-CD of the present invention with different degrees of substitution were selected to examine the product. Acute toxicity, the results showed that the mice were intragastrically administered. When the sample dose reached SOOO mg'kg- 1 , no test animals died and no obvious side effects were observed. When the dose was 2000 mg^g- 1 administered intravenously, no test animals died and no obvious side effects were observed. This experiment failed to measure the LD 5 «) of the two routes of administration of the selected samples, and the maximum tolerated doses were 8000 mg-kg- 1 and 2000 mg-kg- 1 , respectively. The results suggest that the CM- prepared by the present invention HP-P-CD is very toxic. CM-HP-p-CD acute toxicity
CM2-HP6-P-CD 小鼠 口服 - 8000 CM 2 -HP 6 -P-CD mice orally - 8000
静注 - 2000 Intravenous - 2000
CM3-HP6- -CD 小鼠 口服 - 8000 CM 3 -HP 6 - -CD Mouse Oral - 8000
静注 - 2000 Intravenous - 2000
CM3-HP3-P-CD 小鼠 口服 - 8000 CM 3 -HP 3 -P-CD mice orally - 8000
静注 - 2000 受试动物灌胃静注本发明合成的羧甲基-羟丙基 - β -环糊精不仅未观察到 明显的毒副反应且无动物死亡，也未发现动物呼吸、运动等方面存在任何异常， 显示发明合成的产物对试验动物无明显不良影响。 Intravenous Injection - 2000 Intravenous Injection of the Test Animals The carboxymethyl-hydroxypropyl-β-cyclodextrin synthesized in the present invention not only showed no obvious side effects but no animal death, and no animal breathing, exercise, etc. There were any abnormalities in the aspect, indicating that the product synthesized by the invention had no significant adverse effects on the test animals.
以上详细描述了本发明的较佳具体实施例。应当理解，本领域的普通技术无需 创造性劳动就可以根据本发明的构思作出诸多修改和变化。因此，凡本技术领域中 技术人员依本发明的构思在现有技术的基础上通过逻辑分析、推理或者有限的实验 可以得到的技术方案， 皆应在由权利要求书所确定的保护范围内。
The above has described in detail the preferred embodiments of the invention. It will be appreciated that many modifications and variations can be made in the present invention without departing from the scope of the invention. Therefore, any technical solution that can be obtained by a person skilled in the art based on the prior art by logic analysis, reasoning or limited experimentation should be within the scope of protection determined by the claims.
Claims
1、 一种羧甲基-羟丙基 - e -环糊精， 其特征在于， 所述羧甲基-羟丙基 环糊 精是以羧甲基和羟丙基混合取代的 β-环糊精衍生物： η-(2，3,6-0-羧甲 基) -m-(2,3,6-0-2-羟丙基) -β-环糊精， 该 β-环糊精衍生物具有下列通式所示的结 构： 1. A carboxymethyl-hydroxypropyl-e-cyclodextrin, characterized in that the carboxymethyl-hydroxypropyl cyclodextrin is a β-cyclodextrin substituted with a mixture of carboxymethyl and hydroxypropyl groups Spirit derivatives: eta-(2,3,6-0-carboxymethyl)-m-(2,3,6-0-2-hydroxypropyl)-β-cyclodextrin, the β-cyclodextrin The derivative has the structure shown by the following general formula:
Rl= RrO¾COO-X, Rj= CH2CH(OIOC½ , or Rl= RrO¾COO-X, Rj= CH2CH(OIOC½ , or
X= Ν^ , Η X= Ν^ , Η
2、 按照权利要求 1所述的羧甲基-羟丙基 - β -环糊精， 其特征在于： 该环糊精 衍生物同时具有羧甲基和 2-羟丙基两种取代基，该结构物以 CMn-HPm-p-CD表 示， 其羧甲基和 2-羟丙基是同时连接于环糊精母体形成的醚键取代衍生物。2. Carboxymethyl-hydroxypropyl-β-cyclodextrin according to claim 1, characterized in that: the cyclodextrin derivative has both carboxymethyl and 2-hydroxypropyl substituents, and the The structure is represented by CM n -HP m -p-CD, and its carboxymethyl group and 2-hydroxypropyl group are ether bond substituted derivatives formed by simultaneously connecting to the cyclodextrin parent.
3、 按照权利要求 1所述的羧甲基-羟丙基 - 0 -环糊精， 其特征在于： 每摩尔环 糊精取代的基团数有 n个羧甲基和 m个羟丙基； 其中 m为每摩尔环糊精衍生 物中连接的羟丙基取代基个数即羟丙基平均取代度； n为每摩尔环糊精衍生物 中连接的羧甲基取代基个数即羧甲基平均取代度； 其中 m取值为 1~6， n取值 为 1〜6。 3. Carboxymethyl-hydroxypropyl-0-cyclodextrin according to claim 1, characterized in that: the number of groups substituted per mole of cyclodextrin is n carboxymethyl groups and m hydroxypropyl groups; Where m is the number of connected hydroxypropyl substituents per mole of cyclodextrin derivative, that is, the average degree of substitution of hydroxypropyl; n is the number of connected carboxymethyl substituents per mole of cyclodextrin derivative, that is, carboxymethyl The base average degree of substitution; where m takes a value of 1 to 6, and n takes a value of 1 to 6.
4、 按照权利要求 3所述的羧甲基-羟丙基 环糊精， 其特征在于： 环糊精衍 生物的总平均取代度 Z = m + n ， 其中 Z取值为 2~10。 4. The carboxymethyl-hydroxypropyl cyclodextrin according to claim 3, characterized in that: the total average degree of substitution of the cyclodextrin derivatives is Z = m + n, where the value of Z is 2 to 10.
5、 按照权利要求 4所述的羧甲基-羟丙基 - β -环糊精， 其特征在于： 2-羟丙基或 羧甲基取代基的取代位是随机取代于 环糊精葡萄糖单元的 2位， 或 3位， 或 6 位的衍生物。 5. Carboxymethyl-hydroxypropyl-β-cyclodextrin according to claim 4, characterized in that: the substitution position of the 2-hydroxypropyl or carboxymethyl substituent is randomly substituted with the cyclodextrin glucose unit 2-digit, 3-digit, or 6-digit derivatives.
23 twenty three
更正页 （细则第 91条)
6、 按照权利要求 5所述的羧甲基-羟丙基 -β-环糊精， 其特征在于： 所述羧甲 基-羟丙基 -0-环糊精选自： Correction page (Rule 91) 6. Carboxymethyl-hydroxypropyl-β-cyclodextrin according to claim 5, characterized in that: the carboxymethyl-hydroxypropyl-β-cyclodextrin is selected from:
-HPs-P-CD; Z = 6、 -HPs-P-CD; Z = 6,
6、
6.
CM2-HP2-P-CD; Z =4、 CM3-HP4-P-CD; Z = 7、 CM 2 -HP 2 -P-CD; Z =4, CM 3 -HP 4 -P-CD; Z = 7,
CMi-HPs-P-CD; Z =4、 CM2-HP5-P-CD; Z = 7、 CMi-HPs-P-CD; Z =4, CM 2 -HP 5 -P-CD ; Z = 7,
CMi-HP4- -CD; Z =5、 CMi-HP6-P-CD; Z = 7、 CMi-HP 4 - -CD; Z =5, CMi-HP 6 -P-CD; Z = 7,
CM3-HP2- -CD; Z =5、 CM3-HP5-P-CD; Z = 8、
Z =5、 CM2-HP6- -CD; Z = 8、 CM 3 -HP 2 - -CD; Z =5, CM 3 -HP 5 -P-CD ; Z = 8, Z =5, CM 2 -HP 6 - -CD; Z = 8,
CM2-HP3- -CD; Z =5、 CM6-HP3-P-CD; Z = 9或 CM 2 -HP 3 - -CD ; Z =5, CM 6 -HP 3 -P-CD ; Z = 9 or
CM2-HP4- -CD; Z =6、 CMs-HPs-p-CD; Z = 10。 CM 2 -HP 4 - -CD ; Z = 6, CMs-HPs-p-CD; Z = 10.
7、 按照权利要求 6所述的羧甲基-羟丙基 -e-环糊精， 其特征在于： 所述羧甲 基-羟丙基- β -环糊精选自 Ζ = 4〜7的羧甲基-羟丙基- β -环糊精： 7. Carboxymethyl-hydroxypropyl-e-cyclodextrin according to claim 6, characterized in that: the carboxymethyl-hydroxypropyl-β-cyclodextrin is selected from Z=4~7 Carboxymethyl-hydroxypropyl-β-cyclodextrin:
CM2-HP2-p-CD、 CM3-HP2-P-CD, CMi-HP6-P-CD、 CMi-HP3-P-CD、 CMi-HPs-p-CD. CM2-HP5-p-CD、 CMi-HP4-P-CD、 CM2-HP4-p-CD、 CM3-HP4-p-CD、 CM2-HP3- -CD或 CM3-HP3-p-CD。 CM 2 -HP 2 -p-CD, CM3-HP2-P-CD, CMi-HP 6 -P-CD, CMi-HP 3 -P-CD, CMi-HPs-p-CD. CM 2 -HP 5 - p-CD, CMi-HP 4 -P-CD, CM 2 -HP4-p-CD, CM 3 -HP 4 -p-CD, CM 2 -HP 3 - -CD or CM3-HP 3 -p-CD.
8、 按照权利要求 1所述的羧甲基-羟丙基 -P-环糊精，其特征在于：母体 β-环糊 精的含量不超过 1.5% 。 8. Carboxymethyl-hydroxypropyl-P-cyclodextrin according to claim 1, characterized in that: the content of parent β-cyclodextrin does not exceed 1.5%.
9、 一种权利要求 1-8所述的羧甲基-羟丙基 -β-环糊精的制备方法， 其特征在 于，釆用碱催化连续反应的方法进行制备，具体反应步骤是以下两种方案之一: 方案 1: 取 Ρ -环糊精加入 β -环糊精 2.2倍质量的水和 7.0倍摩尔量的碱， 搅泮溶解，室温下搅拌滴加 7.0倍摩尔量的 1，2-环氧丙烷反应，数小时后补加 8.0 倍摩尔量的碱， 加热， 升温稳定后加入 4.0摩尔量的氯乙酸乙酯， 反应生成特 定取代度的羧甲基-羟丙基 -β-环糊精产物； 9. A method for preparing carboxymethyl-hydroxypropyl-β-cyclodextrin according to claims 1-8, characterized in that it is prepared by a base-catalyzed continuous reaction method, and the specific reaction steps are the following two One of the options: Option 1: Take P-cyclodextrin, add 2.2 times the mass of β-cyclodextrin to water and 7.0 times the molar amount of alkali, stir to dissolve, stir and drop 7.0 times the molar amount of 1,2 at room temperature. - Propylene oxide reaction, add 8.0 times the molar amount of alkali after a few hours, heat, add 4.0 molar amount of ethyl chloroacetate after the temperature is stable, and react to generate carboxymethyl-hydroxypropyl-β-ring with a specific degree of substitution Dextrin products;
方案 2: 取 β -环糊精加入 Ρ -环糊精 2.2倍质量的水和 11.0〜35.0倍摩尔量 的碱， 搅拌溶解， 加热下滴加 2.0~10.5倍摩尔量的氯乙酸乙酯反应数小时， 冷 却至室温， 然后控制室温下加入 1.5〜7.0倍摩尔量的 1,2-环氧丙烷， 反应数小时 即生成特定取代度的羧甲基-羟丙基- β -环糊精产物。 Scheme 2: Take β-cyclodextrin, add 2.2 times the mass of water to P-cyclodextrin and 11.0~35.0 times the molar amount of alkali, stir to dissolve, and add 2.0~10.5 times the molar amount of ethyl chloroacetate dropwise under heating. hours, cool to room temperature, and then add 1.5 to 7.0 times the molar amount of 1,2-epoxypropane under controlled room temperature, and react for several hours to generate a carboxymethyl-hydroxypropyl-β-cyclodextrin product with a specific degree of substitution.
10、 按照权利要求 9所述的方法， 其特征在于： 以母体 环糊精为原料， 采 10. The method according to claim 9, characterized in that: using parent cyclodextrin as raw material,
24 twenty four
更正页 （细则第 91条）
用反应中间体不经分离的连续加料反应步骤进行混合取代。 Correction page (Rule 91) Mixing and substitution are carried out using continuous feeding reaction steps without separation of reaction intermediates.
11、 权利要求 1-8中任一项所述的羧甲基-羟丙基 环糊精在药学上的应用。 11. The pharmaceutical application of the carboxymethyl-hydroxypropyl cyclodextrin according to any one of claims 1 to 8.
12、 按照权利要求 11所述的应用， 其特征在于， 所述的羧甲基-羟丙基 - e - 环糊精在药学上用作为药物辅料。 12. The application according to claim 11, characterized in that the carboxymethyl-hydroxypropyl-e-cyclodextrin is used as a pharmaceutical excipient in medicine.
13、 按照权利要求 12所述的应用， 其特征在于， 所述的羧甲基-羟丙基 - β - 环糊精在药学上用作为赋形剂。 13. The application according to claim 12, characterized in that the carboxymethyl-hydroxypropyl-β-cyclodextrin is used as an excipient in medicine.
14、 一种药物组合物， 其特征在于， 所述药物组合物含有权利要求 1-8任一项 所述的羧甲基-羟丙基 环糊精。 14. A pharmaceutical composition, characterized in that the pharmaceutical composition contains the carboxymethyl-hydroxypropyl cyclodextrin described in any one of claims 1-8.
25 25
更正页 （细则第 91条)
Correction Page (Rule 91)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310439451.0 | 2013-09-25 | ||
| CN201310439451.0A CN103554307B (en) | 2013-09-25 | 2013-09-25 | carboxymethyl-hydroxypropyl-beta-cyclodextrin and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2015042759A1 true WO2015042759A1 (en) | 2015-04-02 |
Family
ID=50008691
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/CN2013/001333 WO2015042759A1 (en) | 2013-09-25 | 2013-11-04 | Carboxymethyl-hydroxypropyl-β-cyclodextrin and preparation method thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN103554307B (en) |
| WO (1) | WO2015042759A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018140856A1 (en) | 2017-01-30 | 2018-08-02 | Bio-Rad Laboratories, Inc. | Emulsion compositions and methods of their use |
| FR3075799A1 (en) * | 2017-12-22 | 2019-06-28 | Roquette Freres | PROCESS FOR MODIFICATION OF POLYSACCHARIDE MATERIAL BY HOMOGENEOUS CHEMICAL FUNCTIONALIZATION SEQUENCED |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104558253A (en) * | 2014-12-19 | 2015-04-29 | 福建工程学院 | Green synthesis method of 2-O-methyl-6-O-(2-hydroxypropyl)-beta-cyclodextrin |
| CN104983784A (en) * | 2015-07-30 | 2015-10-21 | 淄博千汇生物科技有限公司 | Method for increasing solubility of pueraria flavonoids |
| CN107573427B (en) * | 2017-10-19 | 2019-10-15 | 无锡甜丰食品有限公司 | A kind of preparation method of hydroxypropyl maltodextrin |
| CN107540750B (en) * | 2017-10-19 | 2019-10-15 | 无锡甜丰食品有限公司 | A kind of preparation method of carboxymethyl maltodextrin |
| CN108503729A (en) * | 2018-03-15 | 2018-09-07 | 山东滨州智源生物科技有限公司 | A kind of ion exchange resin is used to produce the preparation method of HYDROXYPROPYL BETA-CYCLODEXTRIN as catalyst |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4582900A (en) * | 1983-12-17 | 1986-04-15 | Hoechst Aktiengesellschaft | Water-soluble mixed ethers of β-cyclodextrin and a process for their preparation |
| WO1992014762A1 (en) * | 1991-02-15 | 1992-09-03 | Janssen Pharmaceutica N.V. | (carboxyl)alkyloxyalkyl derivatives of cyclodextrins |
| CN1550505A (en) * | 1999-12-16 | 2004-12-01 | 伊斯曼化学公司 | Cyclodextrin ethers |
| WO2007082897A2 (en) * | 2006-01-17 | 2007-07-26 | B. Braun Melsungen Ag | Polysaccharide-cyclodextrin conjugates |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4440236A1 (en) * | 1994-11-10 | 1996-05-15 | Wacker Chemie Gmbh | Redispersible polymer powder composition containing cyclodextrins or cyclodextrin derivatives |
| CN1166693C (en) * | 2001-06-18 | 2004-09-15 | 中奇制药技术（石家庄）有限公司 | Butylbenzene phthalein cyclodextrin or cyclodextrin derivative clathrate, its preparation method and application |
| CA2543443A1 (en) * | 2003-10-31 | 2005-05-12 | The University Of Kansas | Sulfoalkyl ether-alkyl ether cyclodextrin derivatives |
| CN1663968A (en) * | 2005-02-21 | 2005-09-07 | 山东大学 | Hydroxy butyl cyclodextrin derivatives and method for preparing same |
-
2013
- 2013-09-25 CN CN201310439451.0A patent/CN103554307B/en not_active Expired - Fee Related
- 2013-11-04 WO PCT/CN2013/001333 patent/WO2015042759A1/en active Application Filing
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4582900A (en) * | 1983-12-17 | 1986-04-15 | Hoechst Aktiengesellschaft | Water-soluble mixed ethers of β-cyclodextrin and a process for their preparation |
| WO1992014762A1 (en) * | 1991-02-15 | 1992-09-03 | Janssen Pharmaceutica N.V. | (carboxyl)alkyloxyalkyl derivatives of cyclodextrins |
| CN1550505A (en) * | 1999-12-16 | 2004-12-01 | 伊斯曼化学公司 | Cyclodextrin ethers |
| WO2007082897A2 (en) * | 2006-01-17 | 2007-07-26 | B. Braun Melsungen Ag | Polysaccharide-cyclodextrin conjugates |
Non-Patent Citations (1)
| Title |
|---|
| ZHANG, LIEFENG ET AL.: "Synthesis and Evaluation of a Novel beta-Cyclodextrin Derivative for Oral Insulin Delivery and Absorption.", INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES., vol. 61, 28 August 2013 (2013-08-28), pages 494 - 500 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018140856A1 (en) | 2017-01-30 | 2018-08-02 | Bio-Rad Laboratories, Inc. | Emulsion compositions and methods of their use |
| FR3075799A1 (en) * | 2017-12-22 | 2019-06-28 | Roquette Freres | PROCESS FOR MODIFICATION OF POLYSACCHARIDE MATERIAL BY HOMOGENEOUS CHEMICAL FUNCTIONALIZATION SEQUENCED |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103554307B (en) | 2017-01-11 |
| CN103554307A (en) | 2014-02-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2015042759A1 (en) | Carboxymethyl-hydroxypropyl-β-cyclodextrin and preparation method thereof | |
| JP6956767B2 (en) | Production method for cyclodextrin derivatives | |
| US8278437B2 (en) | Hydroxypropyl-sulfobutyl-beta-cyclodextrin, the preparation method, the analytical method, and the pharmacutical application thereof | |
| JP2722277B2 (en) | Highly water-soluble cyclodextrin derivative composition and use thereof | |
| JP4820758B2 (en) | Novel block copolymer, micelle preparation, and anticancer agent containing the same as an active ingredient | |
| US20150238634A1 (en) | Inclusion Compounds of Fumagillol Derivative or its Salt, and Pharmaceutical Compositions Comprising the Same | |
| KR20050013548A (en) | Complex of organic medicines and beta-cyclodextrin derivatives and its preparing process | |
| JP4757633B2 (en) | Micellar preparation containing poorly water-soluble anticancer agent and novel block copolymer | |
| US20080318898A1 (en) | Inclusion complexes of butylphthalide with cyclodextrin or its derivatives, a process for their preparation and the use thereof | |
| EP2080524A1 (en) | Pharmaceutical composition containing docetaxel-cyclodextrin inclusion complex and its preparing process | |
| Khan et al. | Cyclodextrin: an overview | |
| CN109620799B (en) | Stable bromhexine hydrochloride liquid preparation composition and preparation method thereof | |
| CN108239185A (en) | A kind of inclusion compound of quinindium and amine cyclodextrin | |
| KR101128450B1 (en) | Composition comprising ß-cyclodextrin derivatives as a stabilizing agent | |
| CN102861342B (en) | Scutellarin prodrug using cyclodextrin as carrier and preparation method for scutellarin prodrug | |
| JP2021176932A (en) | Carboxylated degradable polyrotaxane and method for producing the same | |
| Cheng et al. | Diblock copolymer glyco-nanomicelles constructed by a maltoheptaose-based amphiphile for reduction-and pH-mediated intracellular drug delivery | |
| CN110156736B (en) | Daidzein carbamate prodrug, salt thereof, preparation method and application thereof | |
| CN108096584B (en) | By using supercritical CO2Tosufloxacin tosylate/cyclodextrin inclusion compound prepared by technology, preparation method and oral preparation thereof | |
| CN111346234A (en) | Deferoxamine-eight-arm star-shaped polyethylene glycol conjugate and application thereof | |
| CN113087814B (en) | Amphiphilic xylan carrier and preparation method thereof | |
| CN109265676B (en) | Folate polyethylene glycol cholesterol lipid material and application thereof | |
| CN115869312B (en) | PDC anti-tumor medicine and preparation method and application thereof | |
| WO2015002078A1 (en) | Novel boronic acid compound preparation | |
| CN117357664A (en) | Clathrate compound of methyl-beta-cyclodextrin and albendazole and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 13894176 Country of ref document: EP Kind code of ref document: A1 |
|
| NENP | Non-entry into the national phase |
Ref country code: DE |
|
| 122 | Ep: pct application non-entry in european phase |
Ref document number: 13894176 Country of ref document: EP Kind code of ref document: A1 |
|
| 32PN | Ep: public notification in the ep bulletin as address of the adressee cannot be established |
Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC |
|
| 122 | Ep: pct application non-entry in european phase |
Ref document number: 13894176 Country of ref document: EP Kind code of ref document: A1 |