CN114478622A - Preparation method of phosphatidylcholine - Google Patents
Preparation method of phosphatidylcholine Download PDFInfo
- Publication number
- CN114478622A CN114478622A CN202210263401.0A CN202210263401A CN114478622A CN 114478622 A CN114478622 A CN 114478622A CN 202210263401 A CN202210263401 A CN 202210263401A CN 114478622 A CN114478622 A CN 114478622A
- Authority
- CN
- China
- Prior art keywords
- acid
- phosphatidylcholine
- lysophosphatidylcholine
- reaction
- crude product
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 title claims abstract description 38
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 238000005917 acylation reaction Methods 0.000 claims abstract description 78
- 239000003960 organic solvent Substances 0.000 claims abstract description 30
- 235000014113 dietary fatty acids Nutrition 0.000 claims abstract description 29
- 229930195729 fatty acid Natural products 0.000 claims abstract description 29
- 239000000194 fatty acid Substances 0.000 claims abstract description 29
- -1 (S) -glycidol fatty acid ester Chemical class 0.000 claims abstract description 28
- 238000007142 ring opening reaction Methods 0.000 claims abstract description 26
- RYCNUMLMNKHWPZ-SNVBAGLBSA-N 1-acetyl-sn-glycero-3-phosphocholine Chemical compound CC(=O)OC[C@@H](O)COP([O-])(=O)OCC[N+](C)(C)C RYCNUMLMNKHWPZ-SNVBAGLBSA-N 0.000 claims abstract description 25
- CTKINSOISVBQLD-GSVOUGTGSA-N (R)-Glycidol Chemical compound OC[C@@H]1CO1 CTKINSOISVBQLD-GSVOUGTGSA-N 0.000 claims abstract description 23
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 21
- YHHSONZFOIEMCP-UHFFFAOYSA-O phosphocholine Chemical compound C[N+](C)(C)CCOP(O)(O)=O YHHSONZFOIEMCP-UHFFFAOYSA-O 0.000 claims abstract description 21
- 229950004354 phosphorylcholine Drugs 0.000 claims abstract description 21
- 238000000034 method Methods 0.000 claims abstract description 20
- 230000010933 acylation Effects 0.000 claims abstract description 19
- 150000001732 carboxylic acid derivatives Chemical class 0.000 claims description 19
- 238000002156 mixing Methods 0.000 claims description 19
- 239000003795 chemical substances by application Substances 0.000 claims description 18
- 150000008065 acid anhydrides Chemical class 0.000 claims description 16
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 claims description 16
- VKOBVWXKNCXXDE-UHFFFAOYSA-N icosanoic acid Chemical compound CCCCCCCCCCCCCCCCCCCC(O)=O VKOBVWXKNCXXDE-UHFFFAOYSA-N 0.000 claims description 16
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 13
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 claims description 12
- 150000004665 fatty acids Chemical class 0.000 claims description 11
- NGNBDVOYPDDBFK-UHFFFAOYSA-N 2-[2,4-di(pentan-2-yl)phenoxy]acetyl chloride Chemical compound CCCC(C)C1=CC=C(OCC(Cl)=O)C(C(C)CCC)=C1 NGNBDVOYPDDBFK-UHFFFAOYSA-N 0.000 claims description 10
- 239000003054 catalyst Substances 0.000 claims description 10
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 claims description 10
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 claims description 10
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 claims description 8
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 claims description 8
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 claims description 8
- 239000005642 Oleic acid Substances 0.000 claims description 8
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 claims description 8
- 150000001263 acyl chlorides Chemical class 0.000 claims description 8
- 229940114079 arachidonic acid Drugs 0.000 claims description 8
- 235000021342 arachidonic acid Nutrition 0.000 claims description 8
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 claims description 8
- WTBAHSZERDXKKZ-UHFFFAOYSA-N octadecanoyl chloride Chemical compound CCCCCCCCCCCCCCCCCC(Cl)=O WTBAHSZERDXKKZ-UHFFFAOYSA-N 0.000 claims description 8
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 claims description 8
- 239000007787 solid Substances 0.000 claims description 8
- 239000003930 superacid Substances 0.000 claims description 8
- UNSAJINGUOTTRA-UHFFFAOYSA-N 3-(3-bromophenyl)prop-2-yn-1-ol Chemical compound OCC#CC1=CC=CC(Br)=C1 UNSAJINGUOTTRA-UHFFFAOYSA-N 0.000 claims description 7
- OCNZHGHKKQOQCZ-CLFAGFIQSA-N [(z)-octadec-9-enoyl] (z)-octadec-9-enoate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC(=O)CCCCCCC\C=C/CCCCCCCC OCNZHGHKKQOQCZ-CLFAGFIQSA-N 0.000 claims description 7
- NWADXBLMWHFGGU-UHFFFAOYSA-N dodecanoic anhydride Chemical compound CCCCCCCCCCCC(=O)OC(=O)CCCCCCCCCCC NWADXBLMWHFGGU-UHFFFAOYSA-N 0.000 claims description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 239000005639 Lauric acid Substances 0.000 claims description 5
- 235000021314 Palmitic acid Nutrition 0.000 claims description 5
- ARBOVOVUTSQWSS-UHFFFAOYSA-N hexadecanoyl chloride Chemical compound CCCCCCCCCCCCCCCC(Cl)=O ARBOVOVUTSQWSS-UHFFFAOYSA-N 0.000 claims description 5
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 claims description 5
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 claims description 5
- WVJVHUWVQNLPCR-UHFFFAOYSA-N octadecanoyl octadecanoate Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC(=O)CCCCCCCCCCCCCCCCC WVJVHUWVQNLPCR-UHFFFAOYSA-N 0.000 claims description 5
- 229910000314 transition metal oxide Inorganic materials 0.000 claims description 5
- 235000021355 Stearic acid Nutrition 0.000 claims description 4
- 125000004432 carbon atom Chemical group C* 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 claims description 4
- 239000008117 stearic acid Substances 0.000 claims description 4
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 3
- 125000002091 cationic group Chemical group 0.000 claims description 3
- 229910017604 nitric acid Inorganic materials 0.000 claims description 3
- 239000011347 resin Substances 0.000 claims description 3
- 229920005989 resin Polymers 0.000 claims description 3
- 150000002190 fatty acyls Chemical group 0.000 claims description 2
- 230000035484 reaction time Effects 0.000 claims description 2
- 238000006243 chemical reaction Methods 0.000 abstract description 13
- 230000008901 benefit Effects 0.000 abstract description 5
- 230000008569 process Effects 0.000 abstract description 4
- 238000003786 synthesis reaction Methods 0.000 abstract description 3
- 239000002699 waste material Substances 0.000 abstract description 2
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 87
- 239000012043 crude product Substances 0.000 description 81
- 239000011259 mixed solution Substances 0.000 description 54
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 39
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 39
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 38
- 238000001914 filtration Methods 0.000 description 36
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 33
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 33
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 32
- 238000003756 stirring Methods 0.000 description 31
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 30
- 239000000706 filtrate Substances 0.000 description 29
- 238000001816 cooling Methods 0.000 description 28
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 26
- 239000012046 mixed solvent Substances 0.000 description 23
- 239000007788 liquid Substances 0.000 description 22
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 description 21
- 238000010438 heat treatment Methods 0.000 description 20
- 239000002904 solvent Substances 0.000 description 20
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 19
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 18
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 18
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 18
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 18
- 238000001953 recrystallisation Methods 0.000 description 16
- 238000001514 detection method Methods 0.000 description 13
- 239000000243 solution Substances 0.000 description 13
- 239000000126 substance Substances 0.000 description 13
- IQGPMZRCLCCXAG-RUZDIDTESA-N 2-stearoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)O[C@H](CO)COP([O-])(=O)OCC[N+](C)(C)C IQGPMZRCLCCXAG-RUZDIDTESA-N 0.000 description 12
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 12
- 239000012024 dehydrating agents Substances 0.000 description 12
- HXJUTPCZVOIRIF-UHFFFAOYSA-N sulfolane Chemical compound O=S1(=O)CCCC1 HXJUTPCZVOIRIF-UHFFFAOYSA-N 0.000 description 12
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 11
- 238000004811 liquid chromatography Methods 0.000 description 11
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 10
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 10
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 10
- CTKINSOISVBQLD-VKHMYHEASA-N (S)-Glycidol Chemical compound OC[C@H]1CO1 CTKINSOISVBQLD-VKHMYHEASA-N 0.000 description 9
- ASWBNKHCZGQVJV-HSZRJFAPSA-O 1-O-palmitoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@@H](O)COP(O)(=O)OCC[N+](C)(C)C ASWBNKHCZGQVJV-HSZRJFAPSA-O 0.000 description 9
- 239000000203 mixture Substances 0.000 description 9
- VXUOFDJKYGDUJI-UHFFFAOYSA-N (2-hydroxy-3-tetradecanoyloxypropyl) 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCC(=O)OCC(O)COP([O-])(=O)OCC[N+](C)(C)C VXUOFDJKYGDUJI-UHFFFAOYSA-N 0.000 description 7
- NRJAVPSFFCBXDT-HUESYALOSA-N 1,2-distearoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCCCC NRJAVPSFFCBXDT-HUESYALOSA-N 0.000 description 7
- SNKAWJBJQDLSFF-NVKMUCNASA-N 1,2-dioleoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC SNKAWJBJQDLSFF-NVKMUCNASA-N 0.000 description 6
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 6
- XTHFKEDIFFGKHM-UHFFFAOYSA-N Dimethoxyethane Chemical compound COCCOC XTHFKEDIFFGKHM-UHFFFAOYSA-N 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 238000005481 NMR spectroscopy Methods 0.000 description 6
- BWKILASWCLJPBO-UHFFFAOYSA-N (3-dodecanoyloxy-2-hydroxypropyl) 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCC(=O)OCC(O)COP([O-])(=O)OCC[N+](C)(C)C BWKILASWCLJPBO-UHFFFAOYSA-N 0.000 description 5
- CITHEXJVPOWHKC-UUWRZZSWSA-N 1,2-di-O-myristoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCC CITHEXJVPOWHKC-UUWRZZSWSA-N 0.000 description 5
- KILNVBDSWZSGLL-KXQOOQHDSA-N 1,2-dihexadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCC KILNVBDSWZSGLL-KXQOOQHDSA-N 0.000 description 5
- IJFVSSZAOYLHEE-SSEXGKCCSA-N 1,2-dilauroyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCC IJFVSSZAOYLHEE-SSEXGKCCSA-N 0.000 description 5
- 230000002378 acidificating effect Effects 0.000 description 5
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 5
- 229960003724 dimyristoylphosphatidylcholine Drugs 0.000 description 5
- XNWFRZJHXBZDAG-UHFFFAOYSA-N 2-METHOXYETHANOL Chemical compound COCCO XNWFRZJHXBZDAG-UHFFFAOYSA-N 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 description 4
- 125000002252 acyl group Chemical group 0.000 description 4
- 125000000217 alkyl group Chemical group 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- SCYULBFZEHDVBN-UHFFFAOYSA-N 1,1-Dichloroethane Chemical compound CC(Cl)Cl SCYULBFZEHDVBN-UHFFFAOYSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 description 3
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 3
- 150000008064 anhydrides Chemical class 0.000 description 3
- 238000010511 deprotection reaction Methods 0.000 description 3
- 239000002502 liposome Substances 0.000 description 3
- 229940049964 oleate Drugs 0.000 description 3
- 150000003904 phospholipids Chemical class 0.000 description 3
- 125000006239 protecting group Chemical group 0.000 description 3
- 239000008096 xylene Substances 0.000 description 3
- YAMUFBLWGFFICM-PTGWMXDISA-N 1-O-oleoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](O)COP([O-])(=O)OCC[N+](C)(C)C YAMUFBLWGFFICM-PTGWMXDISA-N 0.000 description 2
- HZNVUJQVZSTENZ-UHFFFAOYSA-N 2,3-dichloro-5,6-dicyano-1,4-benzoquinone Chemical compound ClC1=C(Cl)C(=O)C(C#N)=C(C#N)C1=O HZNVUJQVZSTENZ-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- PTLZMJYQEBOHHM-AWEZNQCLSA-N [(2S)-oxiran-2-yl]methyl dodecanoate Chemical compound CCCCCCCCCCCC(=O)OC[C@@H]1CO1 PTLZMJYQEBOHHM-AWEZNQCLSA-N 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-M hexadecanoate Chemical compound CCCCCCCCCCCCCCCC([O-])=O IPCSVZSSVZVIGE-UHFFFAOYSA-M 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 238000009776 industrial production Methods 0.000 description 2
- 239000013067 intermediate product Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000005556 structure-activity relationship Methods 0.000 description 2
- 125000000547 substituted alkyl group Chemical group 0.000 description 2
- 238000006257 total synthesis reaction Methods 0.000 description 2
- NGEZPLCPKXKLQQ-VOTSOKGWSA-N (e)-4-(3-methoxyphenyl)but-3-en-2-one Chemical compound COC1=CC=CC(\C=C\C(C)=O)=C1 NGEZPLCPKXKLQQ-VOTSOKGWSA-N 0.000 description 1
- LEBVLXFERQHONN-UHFFFAOYSA-N 1-butyl-N-(2,6-dimethylphenyl)piperidine-2-carboxamide Chemical compound CCCCN1CCCCC1C(=O)NC1=C(C)C=CC=C1C LEBVLXFERQHONN-UHFFFAOYSA-N 0.000 description 1
- PZNPLUBHRSSFHT-RRHRGVEJSA-N 1-hexadecanoyl-2-octadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)O[C@@H](COP([O-])(=O)OCC[N+](C)(C)C)COC(=O)CCCCCCCCCCCCCCC PZNPLUBHRSSFHT-RRHRGVEJSA-N 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 125000002774 3,4-dimethoxybenzyl group Chemical group [H]C1=C([H])C(=C([H])C(OC([H])([H])[H])=C1OC([H])([H])[H])C([H])([H])* 0.000 description 1
- XFPGRCKILOWFOL-UHFFFAOYSA-N 3-chloro-4-phenylmethoxybenzaldehyde Chemical compound ClC1=CC(C=O)=CC=C1OCC1=CC=CC=C1 XFPGRCKILOWFOL-UHFFFAOYSA-N 0.000 description 1
- YZXBAPSDXZZRGB-DOFZRALJSA-M Arachidonate Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC([O-])=O YZXBAPSDXZZRGB-DOFZRALJSA-M 0.000 description 1
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical class OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 201000009906 Meningitis Diseases 0.000 description 1
- ACYNJBAUKQMZDF-HUDVFFLJSA-N [(2s)-oxiran-2-yl]methyl (5z,8z,11z,14z)-icosa-5,8,11,14-tetraenoate Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(=O)OC[C@@H]1CO1 ACYNJBAUKQMZDF-HUDVFFLJSA-N 0.000 description 1
- ATBOMIWRCZXYSZ-XZBBILGWSA-N [1-[2,3-dihydroxypropoxy(hydroxy)phosphoryl]oxy-3-hexadecanoyloxypropan-2-yl] (9e,12e)-octadeca-9,12-dienoate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(COP(O)(=O)OCC(O)CO)OC(=O)CCCCCCC\C=C\C\C=C\CCCCC ATBOMIWRCZXYSZ-XZBBILGWSA-N 0.000 description 1
- ATHVAWFAEPLPPQ-LNVKXUELSA-N [3-octadecanoyloxy-2-[(z)-octadec-9-enoyl]oxypropyl] 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC ATHVAWFAEPLPPQ-LNVKXUELSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- AWUCVROLDVIAJX-UHFFFAOYSA-N alpha-glycerophosphate Natural products OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229940114078 arachidonate Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 229960000684 cytarabine Drugs 0.000 description 1
- 229940070968 depocyt Drugs 0.000 description 1
- 150000002009 diols Chemical class 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 229940042577 exparel Drugs 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 150000002327 glycerophospholipids Chemical class 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- BXZBGYJQEFZICM-UHFFFAOYSA-N icosanoyl chloride Chemical compound CCCCCCCCCCCCCCCCCCCC(Cl)=O BXZBGYJQEFZICM-UHFFFAOYSA-N 0.000 description 1
- 125000000400 lauroyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000002960 lipid emulsion Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- RAFYDKXYXRZODZ-UHFFFAOYSA-N octanoyl octanoate Chemical compound CCCCCCCC(=O)OC(=O)CCCCCCC RAFYDKXYXRZODZ-UHFFFAOYSA-N 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 238000007363 ring formation reaction Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- RCRYHUPTBJZEQS-UHFFFAOYSA-N tetradecanoyl tetradecanoate Chemical compound CCCCCCCCCCCCCC(=O)OC(=O)CCCCCCCCCCCCC RCRYHUPTBJZEQS-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/06—Phosphorus compounds without P—C bonds
- C07F9/08—Esters of oxyacids of phosphorus
- C07F9/09—Esters of phosphoric acids
- C07F9/10—Phosphatides, e.g. lecithin
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
Abstract
The invention belongs to the technical field of organic synthesis, and particularly relates to a preparation method of phosphatidylcholine. The preparation method of the phosphatidylcholine provided by the invention comprises the following steps: dissolving (R) -glycidol and a first acylation reagent in a first organic solvent, and carrying out a first acylation reaction to obtain (S) -glycidol fatty acid ester; and (S) -glycidyl fatty acid ester, phosphorylcholine and a second organic solvent are mixed for ring-opening reaction to obtain the lysophosphatidylcholine. And dissolving lysophosphatidylcholine and a second acylation reagent in a third organic solvent, and carrying out a second acylation reaction to obtain phosphatidylcholine. The method has the advantages of simple process, few reaction steps, mild reaction conditions and more convenient operation; in addition, the post-treatment method is simple and convenient, the discharge of experimental waste is reduced, and the purity of the prepared phosphatidylcholine is more than 98%, so that the method is suitable for industrial preparation.
Description
Technical Field
The invention belongs to the technical field of organic synthesis, and particularly relates to a preparation method of phosphatidylcholine.
Background
Phosphatidylcholine is widely present in animal and plant bodies, is a basic substance for life activities, has important physiological functions and unique emulsifying property, and has important application in the industries of food, health care products, medicines and the like. The medicine prepared from phosphatidylcholine can be used for preparing high-energy parenteral fluid-fat emulsion, carriers of various anticancer drugs and liposome. For example, distearoylphosphatidylcholine is one of the important excipients for the preparation of liposomes, erucylphosphatidylcholine is used in the slow-release polycarbivacaine liposomes known under the name of Exparel, dioleoylphosphatidylcholine is used in the product known under the name of Depocyt (cytarabine) for the treatment of lymphoma meningitis. Extraction from animal and plant carriers rich in phospholipids is a traditional way of preparing phosphatidylcholine, but the substances obtained in the traditional way are mixtures and do not fully meet the market demand.
In order to obtain high-purity phosphatidylcholine with a single and definite structure-activity relationship, researchers synthesize and prepare phosphatidylcholine by a chemical method. For example, in the literature (Synthesis of a Small Library of Mixed-Acid Phospholipids from D-Mannitols as a Homochiral structural Material chem.pharm.Bull.1999,47(11) 1659) -1663), a benzyl-protected chiral diol is used as a starting Material, two identical or different acyl groups are attached according to the purpose, different acyl groups can be introduced by using different activities of primary and secondary hydroxyl groups or by using selective protecting groups, and finally the protecting groups are removed and coupled with phospholipid head groups; the introduction of the acyl group may utilize the corresponding fatty acid, fatty acid anhydride or more reactive acid chloride. For example, in the Master thesis of Chenguang at northwest university, "study on total synthesis of phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol", glycerol is used as a precursor compound to design a total synthesis route of glycerophospholipid containing the same carbon chain; ketal, 3, 4-dimethoxybenzyl are respectively used as dihydroxy and monohydroxy protecting groups of a glycerol skeleton, and acetic acid aqueous solution and 2, 3-dichloro-5, 6-dicyanobenzoquinone (DDQ) are used as corresponding deprotection reagents; the distearoyl phosphatidylcholine is prepared by the steps of reduction, chlorination, condensation, protection and deprotection of dihydroxyl and monohydroxy, acylation, phosphorylation, head group protection, coupling, deprotection and the like.
However, the existing route for preparing phosphatidylcholine has long steps, complex reaction and high requirements on equipment conditions, and is not suitable for scale-up preparation.
Disclosure of Invention
In view of the above, the invention provides a preparation method of phosphatidylcholine, and the preparation method provided by the invention is simple in process, convenient to operate and easy for industrial production.
In order to solve the technical problem, the invention provides a preparation method of phosphatidylcholine, which comprises the following steps:
dissolving (R) -glycidol and a first acylation reagent in a first organic solvent, and carrying out a first acylation reaction to obtain (S) -glycidol fatty acid ester;
mixing the (S) -glycidyl fatty acid ester, phosphorylcholine and a second organic solvent, and carrying out a ring-opening reaction to obtain lysophosphatidylcholine;
and dissolving lysophosphatidylcholine and a second acylation reagent in a third organic solvent, and carrying out a second acylation reaction to obtain phosphatidylcholine.
Preferably, the first acylating agent and the second acylating agent independently comprise a carboxylic acid, an acid chloride, or an acid anhydride.
Preferably, the number of carbon atoms in the carboxylic acid, the acyl chloride and the anhydride is 8-27 independently.
Preferably, the carboxylic acid comprises n-octanoic acid, lauric acid, palmitic acid, stearic acid, arachidic acid, oleic acid or arachidonic acid;
the acyl chloride comprises fatty acyl chloride, myristoyl chloride, palmitoyl chloride or stearoyl chloride;
the acid anhydride includes fatty acid anhydride, lauric acid anhydride, stearic acid anhydride or oleic acid anhydride.
Preferably, the molar ratio of the (R) -glycidol to the first acylating agent is 1:1 to 1.8;
the temperature of the first acylation reaction is-15-50 ℃; the time of the first acylation reaction is 2-8 h.
Preferably, the molar ratio of the lysophosphatidylcholine to the second acylating agent is 1: 1-1.8;
the temperature of the second acylation reaction is 50-150 ℃; the time of the second acylation reaction is 8-24 h.
Preferably, the molar ratio of the phosphorylcholine to the (S) -glycidyl fatty acid ester is 1: 1-1.5.
Preferably, the ring-opening reaction is carried out under the condition of a catalyst;
the mol ratio of the phosphorylcholine to the catalyst is 1: 0.1-1.
Preferably, the catalyst comprises one or more of transition metal oxide, sulfuric acid, nitric acid, hydrochloric acid, cationic resin and solid super acid.
Preferably, the temperature of the ring-opening reaction is 60-150 ℃; the ring-opening reaction time is 12-48 h.
The invention provides a preparation method of phosphatidylcholine, which comprises the following steps: dissolving (R) -glycidol and a first acylation reagent in a first organic solvent, and carrying out a first acylation reaction to obtain (S) -glycidol fatty acid ester; and (S) -glycidyl fatty acid ester, phosphorylcholine and a second organic solvent are mixed for ring-opening reaction to obtain the lysophosphatidylcholine. And dissolving lysophosphatidylcholine and a second acylation reagent in a third organic solvent, and carrying out a second acylation reaction to obtain phosphatidylcholine. The preparation method provided by the invention has the advantages of simple process, few reaction steps and convenience in operation, and is suitable for industrial production. The preparation method provided by the invention has the following advantages:
(1) the preparation method provided by the invention can prepare the phosphatidylcholine with a single and definite structure, and can prepare the high-purity phosphatidylcholine containing symmetrical or asymmetrical, saturated or unsaturated, substituted or unsubstituted double fatty acid chains according to the structure-activity relationship.
(2) The invention takes common (R) -glycidol, carboxylic acid, acyl chloride and acid anhydride as starting raw materials, the conventional chemical phosphorylcholine is used in the intermediate step, the solvents are common industrial reagents, no further treatment is needed, and the invention has the advantage of easily obtained raw materials.
(3) The method has the advantages of simple process, few reaction steps, mild reaction conditions and more convenient operation; in addition, the post-treatment method is simple and convenient, the discharge of experimental waste is reduced, and the purity of the prepared phosphatidylcholine is more than 98%, so that the method is suitable for industrial preparation.
Drawings
FIG. 1 shows lauroyl lysophosphatidylated substance prepared in example 21H NMR spectrum;
FIG. 2 is a liquid chromatogram of lauroyl lysophosphatidylcholine prepared in example 2;
FIG. 3 shows the preparation of dilauroylphosphatidylcholine in example 21H NMR spectrum;
FIG. 4 is a liquid chromatogram of dilauroyl phosphatidylcholine prepared in example 2;
FIG. 5 shows the preparation of myristoyl lysophosphatidylcholine in example 31H NMR spectrum;
FIG. 6 is a liquid chromatogram of myristoyl lysophosphatidylcholine prepared in example 3;
FIG. 7 shows dimyristoyl phosphatidylcholine prepared in example 31H NMR spectrum;
FIG. 8 is a liquid chromatogram of dimyristoyl phosphatidylcholine prepared in example 3;
FIG. 9 shows the preparation of palmitoyl lysophosphatidylcholine in example 41H NMR spectrum;
FIG. 10 is a liquid chromatogram of palmitoyl lysophosphatidylcholine prepared in example 4;
FIG. 11 shows dipalmitoylphosphatidylcholine prepared in example 41H NMR spectrum;
FIG. 12 is a liquid chromatogram of dipalmitoylphosphatidylcholine prepared in example 4;
FIG. 13 shows stearoyl lysophosphatidylcholine prepared in example 51H NMR chart;
FIG. 14 is a liquid chromatogram of stearoyl lysophosphatidylcholine prepared in example 5;
FIG. 15 shows the preparation of distearoylphosphatidylcholine in example 51H NMR spectrum;
FIG. 16 is a liquid chromatogram of distearoylphosphatidylcholine prepared in example 5;
FIG. 17 shows the preparation of dioleoylphosphatidylcholine prepared in example 71H NMR spectrum;
FIG. 18 is a liquid chromatogram of dioleoylphosphatidylcholine prepared in example 7;
FIG. 19 shows the preparation of 1-stearoyl-2-palmitoyl phosphatidylcholine from example 101H NMR spectrum;
FIG. 20 is a liquid chromatogram of 1-stearoyl-2-palmitoyl phosphatidylcholine prepared in example 10.
Detailed Description
The invention provides a preparation method of phosphatidylcholine, which comprises the following steps:
dissolving (R) -glycidol and a first acylation reagent in a first organic solvent, and carrying out a first acylation reaction to obtain (S) -glycidol fatty acid ester;
and (S) -glycidyl fatty acid ester, phosphorylcholine and a second organic solvent are mixed for ring-opening reaction to obtain the lysophosphatidylcholine.
And dissolving lysophosphatidylcholine and a second acylation reagent in a third organic solvent, and carrying out a second acylation reaction to obtain phosphatidylcholine.
The present invention provides a method for producing (S) -glycidol fatty acid ester by dissolving (R) -glycidol and a first acylating agent in a first organic solvent to carry out a first acylation reaction. In the present invention, the first acylating agent preferably comprises a carboxylic acid, an acid chloride or an acid anhydride, more preferably a carboxylic acid or an acid anhydride. In the invention, the number of carbon atoms in the carboxylic acid, the acyl chloride and the anhydride is preferably 8-27, and more preferably 10-26. In the present invention, the carboxylic acid preferably has a structure represented by the formula a-1; the acid chloride preferably has a structure represented by a-2; the acid anhydride preferably has a structure represented by formula a-3;
wherein R is1Preferably C7-C26 alkyl or substituted alkyl, more preferably C7-C26 alkyl. In the present invention, the carboxylic acid preferably includes fatty acid, n-octanoic acid, lauric acid, palmitic acid, stearic acid, arachidic acid, oleic acid, or arachidonic acid, and more preferably, arachidic acid, oleic acid, or arachidonic acid. In the present invention, the acid chloride preferably includes fatty acid chloride, myristoyl chloride, palmitoyl chloride, or stearoyl chloride, and more preferably myristoyl chloride or stearoyl chloride. In the present invention, the acid anhydride preferably includes fatty acid anhydride, lauric acid anhydride, stearic acid anhydride or oleic acid anhydride, and more preferably lauric acid anhydride or oleic acid anhydride.
In the present invention, the first organic solvent preferably includes one or more of acetonitrile, 1, 4-dioxane, N-dimethylformamide, N-dimethylacetamide, ethylene glycol dimethyl ether, dimethyl sulfoxide, sulfolane, dichloromethane, dichloroethane, toluene and xylene, more preferably 1, 4-dioxane, dichloromethane or toluene. In the present invention, when the first organic solvent is two or more of the above specific substances, the ratio of the specific substances in the present invention is not particularly limited, and any ratio may be used.
In the present invention, the molar ratio of the (R) -glycidol to the first acylating agent is preferably 1:1 to 1.8, and more preferably 1:1 to 1.5.
In the present invention, the ratio of the mass of the (R) -glycidol to the volume of the first organic solvent is preferably 19.6g:200 to 500mL, more preferably 19.6g:320 to 450 mL.
In the present invention, the (R) -glycidol, the first acylating agent, and the mixed solution in which the first acylating agent is dissolved in the first organic solvent preferably further include a dehydrating agent, and the dehydrating agent preferably includes N, N-dicyclohexylcarbodiimide, triethylamine, or 4-dimethylaminopyridine. In the present invention, the mass ratio of the (R) -glycidol to the dehydrating agent is preferably 19.6:0.32 to 55, more preferably 19.6:28 to 54.5. In the present invention, it is preferable that the reaction solution obtained by dissolving (R) -glycidol and the first acylating agent in the first organic solvent is heated to a temperature required for the first acylation reaction, and then the dehydrating agent is added.
The invention has no special requirements on the dissolving mode as long as the dissolving mode can be completely dissolved.
In the invention, the temperature of the first acylation reaction is preferably-15-50 ℃; the time of the first acylation reaction is preferably 2-8 h. In the invention, when the first acylating agent is carboxylic acid, the temperature of the first acylation reaction is preferably 45-50 ℃, and the time of the first acylation reaction is preferably 7-8 h; when the first acylation reagent is acid anhydride, the temperature of the first acylation reaction is preferably 25-30 ℃, and the time of the first acylation reaction is preferably 4-5 h; when the first acylation reagent is acyl chloride, the temperature of the first acylation reaction is preferably-15 to-10 ℃, and the time of the first acylation reaction is preferably 2 to 3 hours. In the present invention, the first acylation reaction is preferably accompanied by stirring. In the present invention, the stirring is not particularly limited as long as the reaction can be sufficiently performed.
In the present invention, the equation of the first acylation reaction is shown in formula 1:
in the present invention, it is preferable that the first acylation reaction further comprises: and (2) cooling the system after the first acylation reaction, filtering, concentrating the filtrate obtained by filtering, and recrystallizing to obtain the (S) -glycidyl fatty acid ester.
In the invention, the temperature of the cooled system is preferably room temperature, and the temperature of the room temperature is preferably 20-35 ℃, and more preferably 25-30 ℃. The cooling mode is not particularly limited, and the required temperature can be achieved. The invention has no special requirements on the filtration, and can adopt the conventional filtration mode in the field. In the present invention, the concentration is preferably concentration under reduced pressure. The present invention does not require any particular step for the concentration under reduced pressure, as long as most of the solvent in the system can be removed. In the present invention, the solvent for recrystallization preferably includes a mixed solution of dichloromethane and methanol, a mixed solution of toluene and isopropanol, a mixed solution of ethyl acetate and n-butanol, or a mixed solution of acetone and n-hexane. In the invention, the mass ratio of the dichloromethane and the methanol in the mixed solution of the dichloromethane and the methanol is preferably 1: 2.8-3.2, and more preferably 1:3. In the present invention, the mass ratio of toluene to isopropyl alcohol in the mixed solution of toluene and isopropyl alcohol is preferably 1:2.8 to 3.2, and more preferably 1:3. In the invention, the mass ratio of ethyl acetate to n-butanol in the mixed solution of ethyl acetate and n-butanol is preferably 1: 2.8-3.2, and more preferably 1:3. In the invention, the mass ratio of acetone to n-hexane in the mixed solution of acetone and n-hexane is preferably 1: 2.8-3.2, and more preferably 1:3. In the invention, the mass ratio of the crude product obtained after concentration to the solvent for recrystallization is preferably 1: 3.8-4.2, and more preferably 1: 4.
After obtaining the (S) -glycidyl fatty acid ester, the invention mixes the (S) -glycidyl fatty acid ester, the phosphorylcholine and a second organic solvent, and carries out ring-opening reaction to obtain the lysophosphatidylcholine. In the present invention, the second organic solvent preferably includes one or more of toluene, N-dimethylformamide, N-dimethylacetamide, methanol, ethanol, isopropanol, N-butanol, t-butanol, ethylene glycol methyl ether, ethylene glycol dimethyl ether, dimethyl sulfoxide and sulfolane, and more preferably ethylene glycol methyl ether, sulfolane or N, N-dimethylformamide. In the present invention, when the second organic solvent includes two or more of the above-mentioned specific substances, the ratio of the specific substances in the present invention is not particularly limited, and any ratio may be used.
In the present invention, the mass ratio of the (S) -glycidyl fatty acid ester to the second organic solvent is preferably 32.8 to 91g:400 to 450mL, and more preferably 32.8 to 90.5g:400 to 420 mL. In the present invention, the molar ratio of the phosphorylcholine to the (S) -glycidyl fatty acid ester is preferably 1:1 to 1.5, and more preferably 1:1.2 to 1.4.
In the present invention, the ring-opening reaction is preferably carried out under catalyst conditions; the catalyst preferably comprises one or more of transition metal oxide, sulfuric acid, nitric acid, hydrochloric acid, cationic resin and solid super acid, and more preferably transition metal oxide, sulfuric acid or solid super acid. In the present invention, the transition metal oxide preferably includes acidic alumina. In the present invention, the solid super acid preferably includes a zirconia-sulfuric acid solid super acid. In the present invention, when the catalyst includes two or more of the above-mentioned specific substances, the ratio of the specific substances in the present invention is not particularly limited, and any ratio may be used.
In the present invention, the molar ratio of the phosphorylcholine to the catalyst is preferably 1:0.1 to 1, and more preferably 1:0.2 to 0.8.
The invention has no special requirement on the mixing as long as the uniform mixing can be realized. In the invention, the temperature of the ring-opening reaction is preferably 60-150 ℃, and more preferably 70-140 ℃; the time of the ring-opening reaction is preferably 12-48 h, and more preferably 15-40 h.
In the present invention, the ring-opening reaction is preferably accompanied by stirring, and the stirring is not particularly required in the present invention as long as the reaction can be sufficiently performed.
In the present invention, the formula of the ring-opening reaction is shown in formula 2:
in the present invention, it is preferable that the ring-opening reaction further includes: and (3) cooling the system after the ring-opening reaction, filtering, concentrating the filtrate obtained by filtering, and recrystallizing to obtain the lysophosphatidylcholine.
In the invention, the temperature of the cooled system is preferably room temperature, and the temperature of the room temperature is preferably 20-35 ℃, and more preferably 25-30 ℃. The cooling mode is not particularly limited, and the required temperature can be achieved. The invention has no special requirements on the filtration, and can adopt the conventional filtration mode in the field. In the present invention, the concentration is preferably concentration under reduced pressure. The present invention does not require any particular step for the concentration under reduced pressure, as long as most of the solvent in the system can be removed. In the present invention, the solvent for recrystallization preferably includes a mixture of ethyl acetate and n-hexane, a mixture of chloroform and diethyl ether, or a mixture of acetonitrile and methyl t-butyl ether, and more preferably a mixture of ethyl acetate and n-hexane or a mixture of chloroform and diethyl ether. In the present invention, when the recrystallization solvent is a mixed solution of ethyl acetate and n-hexane, the mass ratio of ethyl acetate to n-hexane is preferably 0.8 to 1.2:1, and more preferably 1: 1; the recrystallization temperature is preferably-1 to 1 ℃, and more preferably 0 ℃. In the invention, when the solvent for recrystallization is a mixed solution of chloroform and diethyl ether, the mass ratio of the chloroform to the diethyl ether is preferably 2.8-3.2: 5, and more preferably 3: 5; the recrystallization temperature is preferably 38-42 ℃, and more preferably 40 ℃. In the invention, when the solvent for recrystallization is a mixed solution of acetonitrile and methyl tert-butyl ether, the mass ratio of the acetonitrile to the methyl tert-butyl ether is preferably 1.8-2.2: 6, and more preferably 2: 6; the temperature of the recrystallization is preferably-42 to-38 ℃, more preferably-40 ℃. In the invention, the mass ratio of the crude product obtained after concentration to the solvent for recrystallization is preferably 1: 7.8-8.2, and more preferably 1: 8. In the present invention, the number of recrystallization is preferably 1 to 5, and more preferably 2 to 4.
After obtaining the lysophosphatidylcholine, the lysophosphatidylcholine and a second acylation reagent are dissolved in a third organic solvent to carry out a second acylation reaction, so that the phosphatidylcholine is obtained. In the present invention, the second acylating agent preferably comprises a carboxylic acid, an acid chloride or an acid anhydride, more preferably a carboxylic acid or an acid anhydride. In the invention, the number of carbon atoms in the carboxylic acid, the acyl chloride and the anhydride is preferably 8-27 independently, and more preferably 10-16 independently. In the present invention, the carboxylic acid preferably has a structure represented by the formula b-1; the acid chloride preferably has a structure represented by b-2; the acid anhydride preferably has a structure represented by the formula b-3;
wherein R is2Preferably C7-C26 alkyl or substituted alkyl, more preferably C7-C26 alkyl. In the present invention, the carboxylic acid preferably includes fatty acid, n-octanoic acid, lauric acid, palmitic acid, stearic acid, arachidic acid, oleic acid, or arachidonic acid, and more preferably arachidic acid, oleic acid, or arachidonic acid. In the present invention, the acid chloride preferably includes fatty acid chloride, myristoyl chloride, palmitoyl chloride, or stearoyl chloride, and more preferably myristoyl chloride or stearoyl chloride. In the present invention, the acid anhydride preferably includes fatty acid anhydride, lauric acid anhydride, stearic acid anhydride or oleic acid anhydride, and more preferably lauric acid anhydride or oleic acid anhydride.
In the present invention, the acyl groups provided by the first and second acylating agents are preferably not identical, i.e. R1And R2Are not identical groups.
In the present invention, the third organic solvent preferably includes one or more of acetonitrile, 1, 4-dioxane, N-dimethylformamide, N-dimethylacetamide, ethylene glycol dimethyl ether, dimethyl sulfoxide, sulfolane, dichloromethane, dichloroethane, and toluene and xylene, more preferably toluene, sulfolane or dimethyl sulfoxide, and still more preferably toluene or dimethyl sulfoxide.
In the present invention, the molar ratio of the lysophosphatidylcholine to the second acylating agent is preferably 1:1 to 1.8, and more preferably 1:1.2 to 1.6.
In the invention, the volume ratio of the mass of the phosphatidylcholine to the third organic solvent is preferably 115-150 g:500mL, and more preferably 120-145 g:500 mL.
In the present invention, the mixed solution of the lysophosphatidylcholine and the second acylating agent dissolved in the third organic solvent preferably further includes a dehydrating agent, and the dehydrating agent preferably includes N, N-Dicyclohexylcarbodiimide (DCC), triethylamine, or 4-Dimethylaminopyridine (DMAP). In the invention, the mass ratio of the lysophosphatidylcholine to the dehydrating agent is preferably 115-150: 0.32-55, and more preferably 120-145: 28-55. In the invention, the reaction solution obtained by dissolving lysophosphatidylcholine and a second acylation reagent in a third organic solvent is preferably heated to the temperature required by the second acylation reaction, and then the dehydrating agent is added.
The invention has no special requirements on the dissolving mode as long as the dissolving mode can be completely dissolved.
In the invention, the temperature of the second acylation reaction is preferably 50-150 ℃; the time of the acylation reaction is preferably 8-24 h. In the invention, when the second acylating agent is carboxylic acid, the temperature of the second acylation reaction is preferably 145-150 ℃, and the time of the second acylation reaction is preferably 22-24 h; when the second acylation reagent is acid anhydride, the temperature of the second acylation reaction is preferably 100-110 ℃, and the time of the second acylation reaction is preferably 14-16 h; when the second acylation reagent is acyl chloride, the temperature of the second acylation reaction is preferably 50-60 ℃, and the time of the second acylation reaction is preferably 8-10 h. In the present invention, the second acylation reaction is preferably accompanied by stirring. In the present invention, the stirring is not particularly limited as long as the reaction can be sufficiently performed.
In the present invention, the equation of the second acylation reaction is shown in formula 3:
in the present invention, it is preferable that the second acylation reaction further comprises: and cooling the system after the second acylation reaction, filtering, concentrating the filtrate obtained by filtering, and recrystallizing to obtain the phosphatidylcholine.
In the invention, the temperature of the cooled system is preferably room temperature, and the temperature of the room temperature is preferably 20-35 ℃, and more preferably 25-30 ℃. The cooling mode is not particularly limited, and the required temperature can be achieved. The invention has no special requirements on the filtration, and can adopt the conventional filtration mode in the field. In the present invention, the concentration is preferably concentration under reduced pressure. The present invention does not require any particular step for the concentration under reduced pressure, as long as most of the solvent in the system can be removed. In the present invention, the solvent for recrystallization preferably includes a good solvent for phosphatidylcholine and a poor solvent for phosphatidylcholine. In the present invention, the good solvent of phosphatidylcholine preferably includes one or more of acetonitrile, ethyl acetate, dichloromethane, dichloroethane, toluene, xylene, chloroform, methanol, ethanol, n-butanol, isopropanol and tert-butanol, more preferably toluene, methanol or acetonitrile. In the invention, the mass ratio of the concentrated solution to the good solvent is preferably 2-4: 1, and more preferably 3: 1. In the present invention, the poor solvent of phosphatidylcholine preferably comprises one or more of n-hexane, cyclohexane, acetone, butanone, diethyl ether and methyl tert-butyl ether, more preferably n-hexane, butanone or methyl tert-butyl ether. In the invention, the mass ratio of the concentrated solution to the poor solvent is preferably 4-6: 1, and more preferably 5: 1. In the examples of the present invention, the solvent for recrystallization is specifically a mixture of toluene and methyl ethyl ketone at a mass ratio of 3:5, a mixture of acetonitrile and methyl tert-butyl ether at a mass ratio of 2:6, or a mixture of methanol and n-hexane at a mass ratio of 1:1. In the examples of the present invention, the mass ratio of the concentrated solution to the recrystallization solvent is specifically 1: 8.
In the invention, the recrystallization temperature is preferably-40 ℃, and more preferably 0-20 ℃. In the present invention, the number of recrystallization is preferably 1 to 5, and more preferably 2 to 4.
The invention adopts the reactant with definite structure and single carbon chain length to ensure that the prepared phosphatidylcholine has definite structure, and the asymmetric phosphatidylcholine can be obtained only by adjusting the types of the first acylation reagent and the second acylation reagent.
In order to further illustrate the present invention, the following embodiments are described in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
Dissolving (R) -glycidol (19.6g, 0.264mol) and N-octanoic acid (68.5g, 0.475mol) in 320mL of 1, 4-dioxane, heating to 50 ℃, adding a dehydrating agent N, N-Dicyclohexylcarbodiimide (DCC) (54.5g, 0.264mol) into the dissolved solution, carrying out a first acylation reaction (accompanied by stirring) for 8h, cooling to 25 ℃, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of toluene and isopropanol with the mass ratio of 1:3 to obtain 49.6g of (S) -glycidol n-caprylate;
mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidol n-caprylate (32.8g, 0.164mol) and 400mL of ethylene glycol monomethyl ether, heating to 60 ℃, carrying out ring-opening reaction (with stirring) for 12h, cooling to room temperature, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the crude product with a mixed solution of ethyl acetate and n-hexane in a mass ratio of 1:1 at 0 deg.C for 3 times (the mass ratio of the crude product to the mixed solution is 1:8) to obtain 38.3g of n-octanoyl lysophosphatidylcholine;
dissolving n-octanoyl lysophosphatidylcholine (101.1g, 0.264mol) and n-octanoic anhydride (52.3g, 0.370mol) in toluene (500mL), heating to 100 ℃, adding 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) into the dissolved solution to perform a second acylation reaction (with stirring) for 14h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product with a mixed solution of toluene and butanone at a mass ratio of 3:5 at 0 ℃ (the mass ratio of the crude product to the mixed solution is 1:8) for 3 times to obtain 116.9g of di-n-octanoyl phosphatidylcholine.
Example 2
Dissolving (R) -glycidol (19.6g, 0.264mol) and lauric acid (95.1g, 0.475mol) in 320mL of 1, 4-dioxane, heating to 50 ℃, adding a dehydrating agent N, N-Dicyclohexylcarbodiimide (DCC) (54.5g, 0.264mol) into the dissolved solution, carrying out a first acylation reaction (accompanied by stirring) for 8h, cooling to 25 ℃, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of toluene and isopropanol with the mass ratio of 1:3 to obtain 63.5g of (S) -glycidyl laurate;
mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidyllaurate (42.0g, 0.164mol), sulfuric acid (1.6g, 16.4mmol) and ethylene glycol monomethyl ether (400mL), heating to 60 ℃, carrying out ring-opening reaction (with stirring) for 12h, cooling to 25 ℃, filtering, collecting filtrate, carrying out reduced pressure concentration to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed liquid is 1:8) for 3 times by using the mixed liquid of ethyl acetate and n-hexane with the mass ratio of 1:1 at the temperature of 0 ℃ to obtain 46.8g of lauroyl lysophosphatidylcholine;
lauroyl lysophosphatidylcholine (115.9g, 0.264mol) and lauric anhydride (141.4g, 0370mol) are dissolved in toluene (500mL), after the temperature is raised to 100 ℃, 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) is added into the dissolved solution to carry out a second acylation reaction (with stirring) for 14h, the cooled temperature is 25 ℃, the filtered solution is filtered, the filtered solution is collected and concentrated under reduced pressure to obtain a crude product, and the crude product is recrystallized by using a mixed solution of toluene and butanone in a mass ratio of 3:5 at the temperature of 0 ℃ (the mass ratio of the crude product to the mixed solution is 1:8) for 3 times to obtain 142.6g of dilauroyl phosphatidylcholine.
Example 3
Mixing (R) -glycidol (19.6g, 0.264mol) and dichloromethane (320mL), cooling to-15 ℃, adding triethylamine (28.1g, 0.278mol) into the mixed solution, adding myristoyl chloride (65.2g, 0.264mol) into the mixed solution of (R) -glycidol, dichloromethane and triethylamine through a constant pressure dropping funnel at a constant speed within 1 hour (with stirring), continuing to react for 1 hour (with stirring), filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product by using a mixed solvent of ethyl acetate and n-butanol in a mass ratio of 1:3 to obtain 68.2g of (S) -glycidylmyristate;
mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidylmyristate (69.9g, 0.246mol), acidic alumina (16.6g, 0.164mol) and sulfolane (420mL), heating to 150 ℃, carrying out ring-opening reaction (with stirring) for 48h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and reacting at 40 ℃ in a mass ratio of 3:5, recrystallizing the crude product for 1 time (the mass ratio of the crude product to the mixed solution is 1:8) by using a mixed solution of chloroform and ether to obtain 50.5g of myristoyl lysophosphatidylcholine;
myristoyl lysophosphatidylcholine (123.3g, 0.264mol), myristic anhydride (162.3g, 0.370mol) and toluene (500mL) are mixed, the temperature is raised to 100 ℃, 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) is added to the mixed solution to carry out a second acylation reaction (with stirring) for 14h, the mixed solution is cooled to 25 ℃, the filtration is carried out, the filtrate is collected and concentrated under reduced pressure to obtain a crude product, and the crude product is recrystallized by using a mixed solvent of toluene and butanone in a mass ratio of 3:5 at the temperature of 0 ℃ (the mass ratio of the crude product to the mixed solution is 1:8) for 3 times to obtain 148.3g of dimyristoyl phosphatidylcholine.
Example 4
Dissolving (R) -glycidol (19.6g, 0.264mol) and palmitic acid (121.8g, 0.475mol) in 320mL of 1, 4-dioxane, heating to 50 ℃, adding a dehydrating agent N, N-Dicyclohexylcarbodiimide (DCC) (54.5g, 0.264mol) into the dissolved solution, carrying out a first acylation reaction (accompanied by stirring) for 8h, cooling to 25 ℃, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of toluene and isopropanol with the mass ratio of 1:3 to obtain 73.3g of (S) -glycidol palmitate;
mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidol palmitate (76.8g, 0.246mol), acidic alumina (16.6g, 0.164mol) and sulfolane (420mL), heating to 150 ℃, carrying out ring-opening reaction (with stirring) for 48h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and reacting at 40 ℃ in a mass ratio of 3:5, recrystallizing the crude product for 1 time by using a mixed solution of chloroform and diethyl ether (the mass ratio of the crude product to the mixed solution is 1:8) to obtain 56.8g of palmitoyl lysophosphatidylcholine;
mixing palmitoyl lysophosphatidylcholine (123.3g, 0.264mol), palmitic anhydride (183.1g, 0.370mol) and toluene (500mL), heating to 100 ℃, adding 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) to the mixed solution to perform a second acylation reaction (with stirring) for 14h, cooling to 25 ℃, filtering, collecting the filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8)1 time at 40 ℃ by using a mixed solvent of toluene and butanone in a mass ratio of 3:5 to obtain 164.5g dipalmitoyl phosphatidylcholine.
Example 5
Dissolving (R) -glycidol (19.6g, 0.264mol) and triethylamine (28.1g, 0.278mol) in 320mL of dichloromethane, cooling to-15 ℃, dropwise adding stearoyl chloride (80.0g, 0.264mol) within 1h at a constant speed, reacting for 1h while keeping the temperature, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of methanol and dichloromethane with the mass ratio of 1:3 to obtain 82.5g of (S) -glycidol stearate;
mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycide stearate (83.6g, 0.246mol), acidic alumina (16.6g, 0.164mol) and sulfolane (420mL), heating to 150 ℃, carrying out cyclization reaction (stirring) for 48h, cooling to 25 ℃, filtering, collecting filtrate, carrying out reduced pressure concentration to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8) by using a mixed solution of chloroform and diethyl ether with the mass ratio of 3:5 at the temperature of 40 ℃ to obtain 59.2g stearoyl lysophosphatidylcholine;
stearoyl lysophosphatidylcholine (138.1g, 0.264mol), stearic anhydride (183.1g, 0.370mol) and toluene (500mL) are mixed, heated to 100 ℃, 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) is added to the mixed solution to carry out a second acylation reaction (with stirring) for 14h, cooled to 25 ℃, filtered, the filtrate is collected and concentrated under reduced pressure to obtain a crude product, and the crude product is recrystallized (the mass ratio of the crude product to the mixed solution is 1:8)1 time by using a mixed solvent of toluene and butanone in a mass ratio of 3:5 at the temperature of 40 ℃ to obtain 183.3g distearoyl phosphatidylcholine.
Example 6
Dissolving (R) -glycidol (19.6g, 0.264mol) and triethylamine (28.1g, 0.278mol) in 320mL of dichloromethane, cooling to-15 ℃, dropwise adding arachidylchloride (87.4g, 0.264mol) within 1h at a constant speed, reacting for 1h while keeping the temperature, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of methanol and dichloromethane with the mass ratio of 1:3 to obtain 92.3g of (S) -glycidic arachidic acid ester;
mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycide arachidic acid ester (90.5g, 0.246mol), acidic alumina (16.6g, 0.164mol) and sulfolane (420mL), heating to 150 ℃, carrying out ring-opening reaction (with stirring) for 48h, cooling to 25 ℃, filtering, collecting filtrate, carrying out reduced pressure concentration to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8)1 time by using a mixed solution of chloroform and diethyl ether with the mass ratio of 3:5 at the temperature of 40 ℃ to obtain 63.3g of arachidoyl lysophosphatidylcholine;
mixing arachidylic lysophosphatidylcholine (145.5g, 0.264mol), arachidic acid (148.4g, 0.475mol) and sulfolane (500mL), heating to 150 ℃, adding N, N-Dicyclohexylcarbodiimide (DCC), 54.5g, 0.264mmol) into the mixed solution, carrying out a second acylation reaction (with stirring) for 24h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8) by using a mixed solvent of toluene and butanone with a mass ratio of 3:5 at 40 ℃ for 1 time to obtain 200.8g of arachidylic acylphosphatidylcholine.
Example 7
Dissolving (R) -glycidol (19.6g, 0.264mol) and oleic anhydride (202.2g, 0.370mol) in toluene (450mL), heating to 30 ℃, adding 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) into the dissolved solution to perform a first acylation reaction (with stirring) for 5h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product of (S) -glycidol oleate, and recrystallizing the crude product at (-5 ℃) by using a mixed solvent of acetone and n-hexane with the mass ratio of 1:3 to obtain 77.7g of (S) -glycidol oleate;
mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidol oleate (66.6g, 0.197mol), zirconia-sulfuric acid solid super acid (10.1g, 0.082mol) and N, N-dimethylformamide (420mL), heating to 100 ℃, carrying out ring-opening reaction (with stirring) for 30h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product with a mixed solution of acetonitrile and methyl tert-butyl ether in a mass ratio of 2:6 (the mass ratio of the crude product to the mixed solution is 1:8) for 5 times at-40 ℃ to obtain 47g of oleoyl lysophosphatidylcholine;
oleoyl lysophosphatidylcholine (137.5g, 0.264mol), oleic acid (134.2g, 0.475mol) and toluene (500mL) are mixed, heated to 150 ℃, 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) is added to the mixed solution to carry out a second acylation reaction (with stirring) for 14h, cooled to 25 ℃, filtered, the filtrate is collected and concentrated under reduced pressure to obtain a crude product, and the crude product is recrystallized (the mass ratio of the crude product to the mixed solution is 1:8)4 times by using a mixed solvent of acetonitrile and methyl tert-butyl ether with the mass ratio of 2:6 at the temperature of-40 ℃ to obtain 149.2g of dioleoyl phosphatidylcholine.
Example 8
Dissolving (R) -glycidol (19.6g, 0.264mol) and arachidonic acid anhydride (218.3g, 0.370mol) in 450mL of toluene, heating to 30 ℃, adding 4-Dimethylaminopyridine (DMAP) (0.32g, 2.64mmol) into a reaction system, preserving heat for reaction for 5h, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of acetone and n-hexane with the mass ratio of 1:3 to obtain 76.2g of (S) -glycidyl arachidonate;
mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidol arachidonate (70.9g, 0.197mol), zirconia-sulfuric acid solid super acid (10.1g, 0.082mol) and N, N-dimethylformamide (400mL), heating to 100 ℃, carrying out ring opening reaction (with stirring) for 30h, cooling to 25 ℃, filtering, collecting filtrate, carrying out reduced pressure concentration to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8) for 5 times by using a mixed solution of acetonitrile and methyl tert-butyl ether with the mass ratio of 2:6 at the temperature of-40 ℃ to obtain 38.3g of arachidonoyl lysophosphatidylcholine;
mixing arachidonic acid lysophosphatidylcholine (143.4g, 0.264mol), arachidonic acid (144.7g, 0.475mol) and sulfolane (500mL), heating to 150 ℃, adding N, N-dicyclohexylcarbodiimide (DCC, 54.5g, 0.264mmol) into the mixed solution, carrying out a second acylation reaction (with stirring) for 24h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8)5 times by using a mixed solvent of acetonitrile and methyl tert-butyl ether with the mass ratio of 2:6 at the temperature of-40 ℃ to obtain 142.4g of the eicosatetraylphosphatidylcholine.
Example 9
Stearoyl lysophosphatidylcholine was prepared according to the method of example 5;
mixing stearoyl lysophosphatidylcholine (138.1g, 0.264mol), oleic acid (134.2g, 0.475mol) and sulfolane (500mL), heating to 150 ℃, adding a dehydrating agent N, N-dicyclohexylcarbodiimide (DCC, 54.5g, 0.264mol) into the mixed solution to perform a second acylation reaction (with stirring) for 24h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8) for 3 times by using a mixed solvent of acetonitrile and methyl tert-butyl ether with the mass ratio of 2:6 at the temperature of-40 ℃ to obtain 153.7g of 1-stearoyl-2-oleoyl phosphatidylcholine.
Example 10
Stearoyl lysophosphatidylcholine was prepared according to the method of example 5;
mixing stearoyl lysophosphatidylcholine (138.1g, 0.264mol) and dimethyl sulfoxide (500mL), heating to 50 ℃, adding triethylamine (28.1g, 0.278mol) into the mixed solution, adding palmitoyl chloride (72.5g, 0.264mol) into the mixed solution of stearoyl lysophosphatidylcholine, dimethyl sulfoxide (500mL) and triethylamine through a constant-pressure dropping funnel within 7 hours, continuing to keep the temperature and stir for 1 hour, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8) for 1 time by using a mixed solvent of methanol and n-hexane with the mass ratio of 1:1 at the temperature of 40 ℃ to obtain 186.8g of 1-stearoyl-2-palmitoyl phosphatidylcholine.
Example 11
Palmitoyl lysophosphatidylcholine was prepared according to the method of example 4;
mixing palmitoyl lysophosphatidylcholine (130.7g, 0.264mol) and dimethyl sulfoxide (500mL), heating to 50 ℃, adding triethylamine (28.1g, 0.278mol) into the mixed solution, adding stearoyl chloride (82.5g, 0.264mol) into the mixed solution of palmitoyl lysophosphatidylcholine, dimethyl sulfoxide and triethylamine through a constant-pressure dropping funnel within 7 hours, continuing to keep the temperature and stir for 1h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8)1 time by using a mixed solvent of methanol and n-hexane with the mass ratio of 1:1 at the temperature of 40 ℃ to obtain 186.8g of 1-palmitoyl-2-stearoyl phosphatidylcholine.
The calculated yields of (S) -glycidyl fatty acid ester, lysophosphatidylcholine and phosphatidylcholine in examples 1 to 11 are shown in Table 1; the purity of lysophosphatidylcholine and phosphatidylcholine in examples 1 to 11 measured by liquid chromatography is shown in table 1.
TABLE 1 yield and purity of intermediate and final products in examples 1-11
Nuclear magnetic detection is carried out on lauroyl lysophosphatidylcholine prepared in example 2 to obtain1The H NMR spectrum is shown in FIG. 1; performing liquid chromatography detection on lauroyl lysophosphatidylcholine prepared in example 2 to obtain a liquid chromatogram shown in FIG. 2; performing nuclear magnetic resonance detection on the dilauroyl phosphatidylcholine prepared in example 2 to obtain1The H NMR spectrum is shown in FIG. 3; the dilauroyl phosphatidylcholine prepared in example 2 was subjected to liquid chromatography, and the liquid chromatogram obtained is shown in fig. 4.
Nuclear magnetic assay of myristoyl lysophosphatidylcholine prepared in example 3 to obtain myristoyl lysophosphatidylcholine1The H NMR spectrum is shown in FIG. 5; performing liquid chromatography detection on myristoyl lysophosphatidylcholine prepared in example 3 to obtain a liquid chromatogram shown in fig. 6; nuclear magnetic resonance detection was performed on dimyristoyl phosphatidylcholine obtained in example 3 to obtain dimyristoyl phosphatidylcholine1The H NMR spectrum is shown in FIG. 7; dimyristoyl phosphatidyl compound prepared in example 3Liquid chromatogram obtained by liquid chromatography detection of choline is shown in FIG. 8.
Subjecting palmitoyl lysophosphatidylcholine obtained in example 4 to nuclear magnetic detection to obtain1The H NMR spectrum is shown in FIG. 9; performing liquid chromatography detection on palmitoyl lysophosphatidylcholine prepared in example 4 to obtain a liquid chromatogram shown in fig. 10; subjecting the dipalmitoylphosphatidylcholine prepared in example 4 to nuclear magnetic resonance detection to obtain1The H NMR spectrum is shown in FIG. 11; the dipalmitoylphosphatidylcholine prepared in example 4 was subjected to liquid chromatography, and the liquid chromatogram obtained was shown in FIG. 12.
Performing nuclear magnetic detection on stearoyl lysophosphatidylcholine prepared in example 5 to obtain stearoyl lysophosphatidylcholine1The H NMR spectrum is shown in FIG. 13; detecting stearoyl lysophosphatidylcholine prepared in example 5 by liquid chromatography, and obtaining a liquid chromatogram as shown in fig. 14; performing nuclear magnetic detection on the distearoyl phosphatidylcholine prepared in example 5 to obtain1The H NMR spectrum is shown in FIG. 15; the distearoyl phosphatidylcholine prepared in example 5 was subjected to liquid chromatography detection, and the liquid chromatogram obtained was shown in fig. 16.
Subjecting the dioleoylphosphatidylcholine prepared in example 7 to nuclear magnetic resonance to obtain1The H NMR spectrum is shown in FIG. 17; the dioleoylphosphatidylcholine prepared in example 7 was subjected to liquid chromatography, and the liquid chromatogram obtained was shown in FIG. 18.
Subjecting the 1-stearoyl-2-palmitoyl phosphatidylcholine obtained in example 10 to nuclear magnetic resonance to obtain1The H NMR spectrum is shown in FIG. 19; the liquid chromatogram obtained by performing liquid chromatography detection on 1-stearoyl-2-palmitoyl phosphatidylcholine prepared in example 10 is shown in fig. 20.
The invention is based on1H NMR spectra are assigned to hydrogen in different chemical environments, so that the structure of the product can be determined, the intermediate product prepared in the embodiment is lysophosphatidylcholine, and the intermediate product is phosphatidylcholine, such as nuclear magnetic resonance hydrogen spectra provided by the invention; meanwhile, according to the retention time of different substances in the high performance liquid chromatogramThe difference in (3) can also be confirmed by comparing with a standard sample to confirm the structure and purity.
Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.
Claims (10)
1. A preparation method of phosphatidylcholine comprises the following steps:
dissolving (R) -glycidol and a first acylation reagent in a first organic solvent, and carrying out a first acylation reaction to obtain (S) -glycidol fatty acid ester;
mixing the (S) -glycidyl fatty acid ester, phosphorylcholine and a second organic solvent, and carrying out a ring-opening reaction to obtain lysophosphatidylcholine;
and dissolving lysophosphatidylcholine and a second acylation reagent in a third organic solvent, and carrying out a second acylation reaction to obtain phosphatidylcholine.
2. The method of claim 1, wherein the first and second acylating reagents independently comprise a carboxylic acid, an acid chloride, or an acid anhydride.
3. The method according to claim 2, wherein the number of carbon atoms in the carboxylic acid, the acid chloride and the acid anhydride is 8 to 27 independently.
4. The production method according to claim 3, wherein the carboxylic acid comprises n-octanoic acid, lauric acid, palmitic acid, stearic acid, arachidic acid, oleic acid or arachidonic acid;
the acyl chloride comprises fatty acyl chloride, myristoyl chloride, palmitoyl chloride or stearoyl chloride;
the acid anhydride includes fatty acid anhydride, lauric acid anhydride, stearic acid anhydride or oleic acid anhydride.
5. The production method according to any one of claims 1 to 4, wherein the molar ratio of the (R) -glycidol to the first acylating agent is 1:1 to 1.8;
the temperature of the first acylation reaction is-15-50 ℃; the time of the first acylation reaction is 2-8 h.
6. The method according to any one of claims 1 to 4, wherein the molar ratio of the lysophosphatidylcholine to the second acylating agent is 1:1 to 1.8;
the temperature of the second acylation reaction is 50-150 ℃; the time of the second acylation reaction is 8-24 h.
7. The method according to claim 1, wherein the molar ratio of the phosphorylcholine to the (S) -glycidyl fatty acid ester is 1:1 to 1.5.
8. The production method according to claim 1, wherein the ring-opening reaction is carried out under a catalyst condition;
the mol ratio of the phosphorylcholine to the catalyst is 1: 0.1-1.
9. The method of claim 8, wherein the catalyst comprises one or more of transition metal oxide, sulfuric acid, nitric acid, hydrochloric acid, cationic resin, and solid super acid.
10. The method according to any one of claims 4 and 7 to 9, wherein the ring-opening reaction is carried out at a temperature of 60 to 150 ℃; the ring-opening reaction time is 12-48 h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210263401.0A CN114478622A (en) | 2022-03-17 | 2022-03-17 | Preparation method of phosphatidylcholine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210263401.0A CN114478622A (en) | 2022-03-17 | 2022-03-17 | Preparation method of phosphatidylcholine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114478622A true CN114478622A (en) | 2022-05-13 |
Family
ID=81486859
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210263401.0A Pending CN114478622A (en) | 2022-03-17 | 2022-03-17 | Preparation method of phosphatidylcholine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114478622A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5536855A (en) * | 1994-03-04 | 1996-07-16 | National Starch And Chemical Investment Holding Corporation | Process for preparing glycidyl esters for use in electronics adhesives |
| WO2009128631A2 (en) * | 2008-04-15 | 2009-10-22 | Hwang Soon Ook | Method for preparing racemic or optically active α-glycerophosphorylcholine and derivative thereof |
| KR20100035897A (en) * | 2008-09-29 | 2010-04-07 | 엔자이텍 주식회사 | METHOD FOR PREPARING RACEMIC OR OPICALLY ACTIVE α-PHOSPHATIDYLCHOLINE DERIVATIVES |
| CN102617633A (en) * | 2012-03-01 | 2012-08-01 | 济南康和医药科技有限公司 | Synthesis method of L-alpha-glycerophosphoryl choline |
| KR101233138B1 (en) * | 2012-11-06 | 2013-02-22 | 주식회사 한서켐 | A PROCESS FOR PREPARING L-α-GLYCERYL PHOSPHORYL CHOLINE |
-
2022
- 2022-03-17 CN CN202210263401.0A patent/CN114478622A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5536855A (en) * | 1994-03-04 | 1996-07-16 | National Starch And Chemical Investment Holding Corporation | Process for preparing glycidyl esters for use in electronics adhesives |
| WO2009128631A2 (en) * | 2008-04-15 | 2009-10-22 | Hwang Soon Ook | Method for preparing racemic or optically active α-glycerophosphorylcholine and derivative thereof |
| KR20100035897A (en) * | 2008-09-29 | 2010-04-07 | 엔자이텍 주식회사 | METHOD FOR PREPARING RACEMIC OR OPICALLY ACTIVE α-PHOSPHATIDYLCHOLINE DERIVATIVES |
| CN102617633A (en) * | 2012-03-01 | 2012-08-01 | 济南康和医药科技有限公司 | Synthesis method of L-alpha-glycerophosphoryl choline |
| KR101233138B1 (en) * | 2012-11-06 | 2013-02-22 | 주식회사 한서켐 | A PROCESS FOR PREPARING L-α-GLYCERYL PHOSPHORYL CHOLINE |
Non-Patent Citations (1)
| Title |
|---|
| YUSUF REYHANOGLU等: "Alternative Approach for Synthesizing Polyglycolic Acid Copolymers from C1 Feedstocks and Fatty Ester Epoxides", 《ACS SUSTAINABLE CHEM. ENG. 》, vol. 7, pages 5103 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107266348B (en) | Preparation, structure and the purposes of 4-acetylbiphenyl hydrazone -3- indolecarboxaldehyde Schiff base | |
| CN114478622A (en) | Preparation method of phosphatidylcholine | |
| CN108586250A (en) | A kind of sodium stearyl fumarate auxiliary material and preparation method thereof | |
| CN115043826B (en) | Sinomenine furazan derivatives, and preparation method and application thereof | |
| EP3395808A1 (en) | Pseudo-ceramide compound and preparation method therefor | |
| CN109206469B (en) | Glycyrrhetinic acid derivative and preparation method and application thereof | |
| US20120283198A1 (en) | Halogenated dideoxy sugar derivates, preparation method and application thereof | |
| CN113321634A (en) | Long-chain primary amide chrysin derivative and preparation method and application thereof | |
| CN111253434A (en) | Preparation method of dipalmitoyl phosphatidic acid | |
| CN102336746B (en) | Preparation method and anti-cancer action of novel diacid solanesol alkyl tegafur diesters | |
| CN102482243A (en) | Solvates of 4-acetoxy-2a-benzoyloxy-5ss,20-epoxy-1,7ss,10ss-trihydroxy-9-oxo-tax-11 -en- 13a-yl (2r,3s)-3-tert-butoxycarbonylamino-2-hydroxy-3-phenylpropionate | |
| CN104098524A (en) | 1-m-methoxy benzoyl-3-phenyl-1, 4-dihydro-1,2,4,5-tetrazine and preparation and application thereof | |
| An et al. | Preparation and in vitro bioactivity evaluation of N-heterocyclic-linked dihomooxacalix [4] arene derivatives | |
| CN115594690B (en) | Metformin biotin MetBio, synthesis method thereof and application thereof in nucleic acid drug delivery | |
| CN111518155B (en) | Dammarenediol trimethyl alcohol compound, crystal form A and preparation method thereof | |
| CN114716433B (en) | Crystal form of demethylenetetrahydroberberine hydrochloride and preparation method thereof | |
| CN113149948B (en) | Preparation method and application of 3-methylene cyclopentano [ b ] chromone compound | |
| CN113831373A (en) | Synthesis method of morphine-6-glucuronide dehydrate impurities | |
| CN107827762B (en) | (2, 3-dioleoyl-propyl) trimethyl ammonium chloride and preparation method and application thereof | |
| BR102020026253A2 (en) | PROCESS FOR THE PRODUCTION OF DIMETHYL FUMARATE | |
| CN116041259A (en) | Hydroxychloroquine derivative and preparation method thereof | |
| CN117304029A (en) | 4,16- [2.2] cyclophenyl diester and 4- [2.2] cyclophenyl monoester, one-pot synthesis preparation method and application thereof | |
| CN114671869A (en) | Novel synthesis method of compound Tomivosertib | |
| CN111349230A (en) | PEG-containing chalcone derivative prodrug, synthesis method and application of PEG-containing chalcone derivative prodrug in preparation of non-alcoholic steatohepatitis resistant drugs | |
| EP0147174A2 (en) | Dihydroxybenzaldehyde derivatives as anti-inflammatory agents |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |