CN113943336A - Method for synthesizing cholesterol by taking BA as raw material - Google Patents
Method for synthesizing cholesterol by taking BA as raw material Download PDFInfo
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- CN113943336A CN113943336A CN202111448458.XA CN202111448458A CN113943336A CN 113943336 A CN113943336 A CN 113943336A CN 202111448458 A CN202111448458 A CN 202111448458A CN 113943336 A CN113943336 A CN 113943336A
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- cholesterol
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- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 title claims abstract description 140
- 235000012000 cholesterol Nutrition 0.000 title claims abstract description 70
- 239000002994 raw material Substances 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 37
- 230000002194 synthesizing effect Effects 0.000 title claims abstract description 20
- 238000006722 reduction reaction Methods 0.000 claims abstract description 39
- 238000005984 hydrogenation reaction Methods 0.000 claims abstract description 24
- 238000007239 Wittig reaction Methods 0.000 claims abstract description 18
- 238000006640 acetylation reaction Methods 0.000 claims abstract description 16
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 15
- 230000021736 acetylation Effects 0.000 claims abstract description 3
- 230000003647 oxidation Effects 0.000 claims abstract description 3
- 150000001875 compounds Chemical class 0.000 claims description 142
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 87
- 238000006243 chemical reaction Methods 0.000 claims description 79
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims description 60
- 239000002904 solvent Substances 0.000 claims description 58
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 51
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 41
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 34
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 29
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 28
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 24
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N triphenylphosphine Chemical compound C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 claims description 22
- WETWJCDKMRHUPV-UHFFFAOYSA-N acetyl chloride Chemical compound CC(Cl)=O WETWJCDKMRHUPV-UHFFFAOYSA-N 0.000 claims description 20
- 239000012346 acetyl chloride Substances 0.000 claims description 20
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 18
- 239000003638 chemical reducing agent Substances 0.000 claims description 18
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 claims description 16
- JRNVZBWKYDBUCA-UHFFFAOYSA-N N-chlorosuccinimide Chemical compound ClN1C(=O)CCC1=O JRNVZBWKYDBUCA-UHFFFAOYSA-N 0.000 claims description 16
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 14
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 14
- 239000003054 catalyst Substances 0.000 claims description 13
- LPNYRYFBWFDTMA-UHFFFAOYSA-N potassium tert-butoxide Chemical compound [K+].CC(C)(C)[O-] LPNYRYFBWFDTMA-UHFFFAOYSA-N 0.000 claims description 11
- 239000012279 sodium borohydride Substances 0.000 claims description 9
- 229910000033 sodium borohydride Inorganic materials 0.000 claims description 9
- 239000007800 oxidant agent Substances 0.000 claims description 7
- 230000001590 oxidative effect Effects 0.000 claims description 7
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 6
- GDOPTJXRTPNYNR-UHFFFAOYSA-N methyl-cyclopentane Natural products CC1CCCC1 GDOPTJXRTPNYNR-UHFFFAOYSA-N 0.000 claims description 6
- JRMUNVKIHCOMHV-UHFFFAOYSA-M tetrabutylammonium bromide Chemical compound [Br-].CCCC[N+](CCCC)(CCCC)CCCC JRMUNVKIHCOMHV-UHFFFAOYSA-M 0.000 claims description 6
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims description 5
- JWUJQDFVADABEY-UHFFFAOYSA-N 2-methyltetrahydrofuran Chemical compound CC1CCCO1 JWUJQDFVADABEY-UHFFFAOYSA-N 0.000 claims description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 4
- 230000035484 reaction time Effects 0.000 claims description 4
- MICMHFIQSAMEJG-UHFFFAOYSA-N 1-bromopyrrolidine-2,5-dione Chemical compound BrN1C(=O)CCC1=O.BrN1C(=O)CCC1=O MICMHFIQSAMEJG-UHFFFAOYSA-N 0.000 claims description 2
- XVMSFILGAMDHEY-UHFFFAOYSA-N 6-(4-aminophenyl)sulfonylpyridin-3-amine Chemical compound C1=CC(N)=CC=C1S(=O)(=O)C1=CC=C(N)C=N1 XVMSFILGAMDHEY-UHFFFAOYSA-N 0.000 claims description 2
- IFPHDUVGLXEIOQ-UHFFFAOYSA-N ortho-iodosylbenzoic acid Chemical compound OC(=O)C1=CC=CC=C1I=O IFPHDUVGLXEIOQ-UHFFFAOYSA-N 0.000 claims description 2
- 238000009776 industrial production Methods 0.000 abstract description 9
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- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 40
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 26
- 239000000243 solution Substances 0.000 description 26
- 239000012074 organic phase Substances 0.000 description 22
- 239000007787 solid Substances 0.000 description 22
- 238000004440 column chromatography Methods 0.000 description 18
- NPXOKRUENSOPAO-UHFFFAOYSA-N Raney nickel Chemical compound [Al].[Ni] NPXOKRUENSOPAO-UHFFFAOYSA-N 0.000 description 17
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 17
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 17
- 239000012535 impurity Substances 0.000 description 17
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- 230000000052 comparative effect Effects 0.000 description 12
- 230000014759 maintenance of location Effects 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 11
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 10
- 238000005160 1H NMR spectroscopy Methods 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 10
- 238000002360 preparation method Methods 0.000 description 10
- 239000012046 mixed solvent Substances 0.000 description 9
- 238000010992 reflux Methods 0.000 description 9
- 239000011734 sodium Substances 0.000 description 9
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 8
- 238000012544 monitoring process Methods 0.000 description 7
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- 229920006395 saturated elastomer Polymers 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- GKASDNZWUGIAMG-UHFFFAOYSA-N triethyl orthoformate Chemical compound CCOC(OCC)OCC GKASDNZWUGIAMG-UHFFFAOYSA-N 0.000 description 6
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium on carbon Substances [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 5
- 229940126214 compound 3 Drugs 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- OILXMJHPFNGGTO-UHFFFAOYSA-N (22E)-(24xi)-24-methylcholesta-5,22-dien-3beta-ol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(C)C(C)C)C1(C)CC2 OILXMJHPFNGGTO-UHFFFAOYSA-N 0.000 description 4
- OQMZNAMGEHIHNN-UHFFFAOYSA-N 7-Dehydrostigmasterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CC(CC)C(C)C)CCC33)C)C3=CC=C21 OQMZNAMGEHIHNN-UHFFFAOYSA-N 0.000 description 4
- XYFCBTPGUUZFHI-UHFFFAOYSA-N Phosphine Chemical compound P XYFCBTPGUUZFHI-UHFFFAOYSA-N 0.000 description 4
- HZYXFRGVBOPPNZ-UHFFFAOYSA-N UNPD88870 Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)=CCC(CC)C(C)C)C1(C)CC2 HZYXFRGVBOPPNZ-UHFFFAOYSA-N 0.000 description 4
- LGJMUZUPVCAVPU-UHFFFAOYSA-N beta-Sitostanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 LGJMUZUPVCAVPU-UHFFFAOYSA-N 0.000 description 4
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- 229910000144 sodium(I) superoxide Inorganic materials 0.000 description 4
- HCXVJBMSMIARIN-PHZDYDNGSA-N stigmasterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)/C=C/[C@@H](CC)C(C)C)[C@@]1(C)CC2 HCXVJBMSMIARIN-PHZDYDNGSA-N 0.000 description 4
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- BFDNMXAIBMJLBB-UHFFFAOYSA-N stigmasterol Natural products CCC(C=CC(C)C1CCCC2C3CC=C4CC(O)CCC4(C)C3CCC12C)C(C)C BFDNMXAIBMJLBB-UHFFFAOYSA-N 0.000 description 4
- CZHLPWNZCJEPJB-UHFFFAOYSA-N 1-chloro-3-methylbutane Chemical compound CC(C)CCCl CZHLPWNZCJEPJB-UHFFFAOYSA-N 0.000 description 3
- 241000283690 Bos taurus Species 0.000 description 3
- 238000005481 NMR spectroscopy Methods 0.000 description 3
- ORNBQBCIOKFOEO-YQUGOWONSA-N Pregnenolone Natural products O=C(C)[C@@H]1[C@@]2(C)[C@H]([C@H]3[C@@H]([C@]4(C)C(=CC3)C[C@@H](O)CC4)CC2)CC1 ORNBQBCIOKFOEO-YQUGOWONSA-N 0.000 description 3
- 229940125898 compound 5 Drugs 0.000 description 3
- 229960000249 pregnenolone Drugs 0.000 description 3
- ORNBQBCIOKFOEO-QGVNFLHTSA-N pregnenolone Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 ORNBQBCIOKFOEO-QGVNFLHTSA-N 0.000 description 3
- 238000001953 recrystallisation Methods 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- YXZFFTJAHVMMLF-UHFFFAOYSA-N 1-bromo-3-methylbutane Chemical compound CC(C)CCBr YXZFFTJAHVMMLF-UHFFFAOYSA-N 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 2
- DHKHKXVYLBGOIT-UHFFFAOYSA-N acetaldehyde Diethyl Acetal Natural products CCOC(C)OCC DHKHKXVYLBGOIT-UHFFFAOYSA-N 0.000 description 2
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- 210000000133 brain stem Anatomy 0.000 description 2
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- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 2
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- 229910052703 rhodium Inorganic materials 0.000 description 2
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- MHOVAHRLVXNVSD-UHFFFAOYSA-N rhodium atom Chemical compound [Rh] MHOVAHRLVXNVSD-UHFFFAOYSA-N 0.000 description 2
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- 239000001632 sodium acetate Substances 0.000 description 2
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- PODWXQQNRWNDGD-UHFFFAOYSA-L sodium thiosulfate pentahydrate Chemical compound O.O.O.O.O.[Na+].[Na+].[O-]S([S-])(=O)=O PODWXQQNRWNDGD-UHFFFAOYSA-L 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
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- 235000005282 vitamin D3 Nutrition 0.000 description 2
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- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 1
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- 239000004166 Lanolin Substances 0.000 description 1
- OPGYCKDTPNLLCZ-CHNGHHIASA-N OC[C@H]([C@H]1CC[C@H]2[C@@H]3CCC4=CC(CC[C@]4(C)[C@H]3CC[C@]12C)=O)C.OCC([C@H]1CC[C@H]2[C@@H]3CCC4=CC(CC[C@]4(C)[C@H]3CC[C@]12C)=O)C Chemical compound OC[C@H]([C@H]1CC[C@H]2[C@@H]3CCC4=CC(CC[C@]4(C)[C@H]3CC[C@]12C)=O)C.OCC([C@H]1CC[C@H]2[C@@H]3CCC4=CC(CC[C@]4(C)[C@H]3CC[C@]12C)=O)C OPGYCKDTPNLLCZ-CHNGHHIASA-N 0.000 description 1
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Steroid Compounds (AREA)
Abstract
The invention discloses a method for synthesizing cholesterol by using BA as a raw material, which is characterized in that a plant source raw material 21-hydroxy-20-methyl pregn-4-ene-3-ketone (also called as dideanol or BA) is used as a raw material, the cholesterol is synthesized by steps of oxidation, Wittig reaction, acetylation, reduction, selective hydrogenation reduction and the like, and the purity can reach the standard (> 95%) of commercial grade cholesterol. The raw materials for synthesizing the cholesterol are plant sources, so that the price is low, the safety is high, the risk of pathogenic bacteria and virus infection is avoided, the synthesis method is simple to operate, the yield is high, the side reaction is few, the environment is friendly, the economy is good, and the industrial production is facilitated; solves the safety problem of the prior cholesterol product and the problems of high cost, unfriendly environment and unsuitability for large-scale industrial production in the synthesis technology.
Description
Technical Field
The invention belongs to the technical field of organic chemical synthesis, and relates to a method for synthesizing cholesterol by using a plant source raw material 21-hydroxy-20-methyl pregn-4-ene-3-ketone (also called as di-noralcohol or BA (Bisnoralgolhol)) as a raw material.
Background
Cholesterol, also known as cholesterol, is widely found in animals, most notably in brain tissue. Cholesterol is an essential substance of animal tissue cells, is not only involved in the formation of cell membranes, but also is a raw material for synthesizing bile acid and steroid hormone, in particular a raw material for synthesizing vitamin D3, and is used as a pharmaceutical adjuvant (injection grade) and a food, feed additive and the like. Currently, cholesterol is commercially available from the brain and spinal cord of animals such as pigs, sheep, cattle, etc. or from lanolin. Researches find that the source of a plurality of diseases is animals, and particularly, as infection events such as mad cow disease, streptococcus suis and avian influenza occur, people pay more and more attention to the safety of cholesterol. If the upstream raw material for producing vitamin D3 is cholesterol, the European and American countries do not allow the use of brainstem cholesterol as the raw material for a long time because of epidemic risks, and China also limits the use of the brainstem cholesterol raw material from 7 months and 1 day in 2020. Therefore, there is an urgent need to develop a plant-derived, safe, green method for cholesterol synthesis.
The chemical synthesis of cholesterol has been reported mainly as follows:
(1) cholesterol (CN 1772760A, shown in Scheme 1) was synthesized in a total yield of 61% by 6 steps of reaction using diosgenin as a raw material. The route has relatively high raw material price, complicated steps, and large toxicity and pollution of used reagents, and is not suitable for industrial production.
(2) The stigmasterol degradation product is used as a raw material, and the cholesterol is synthesized with the total yield of 67 percent through 5 steps of reaction, (CN105218610A, shown in the Scheme 2). However, the applicant of the present invention has experimentally shown that the following problems exist in the route: in the first step of the reaction, triethyl orthoformate is adopted to carry out etherification protection on the C-3 carbonyl group, so that the selectivity is poor, and the side chain aldehyde group is easy to react to form an acetal compound (see the 'comparative example I' in the summary of the invention). Thus, the feasibility of this route presents serious problems.
(3) The pregnenolone is used as a raw material, and the total yield of the pregnenolone is 72% to synthesize the cholesterol through 4 steps of reaction, (CN 105218609A, shown in a Scheme 3), the route uses a noble metal rhodium catalyst and a chiral phosphine ligand, and is expensive and not suitable for large-scale industrial production.
(4) Pregnenolone is used as a raw material, cholesterol is synthesized with the total yield of 80% through 2-step reaction, (CN 104961788A, shown in a Scheme 4), the route also uses a noble metal rhodium catalyst and a chiral phosphine ligand, and the cost is high, so that the method is not suitable for large-scale industrial production.
(5) Using stigmasterol as raw material, synthesizing cholesterol with total yield of 68% (CN 105237603A, shown in Scheme 5) by 5-step reaction, wherein O is used in the synthesis process3Higher requirements are put on monitoring reaction and equipment, and the economy and the safety are not good enough.
(6) Using stigmasterol as raw material, synthesizing cholesterol with total yield of 70% by 4-step reaction, (CN 106632565A, shown in Scheme 6), the route also uses O3The process difficulty is increased, higher requirements are put on monitoring reaction and equipment, and the economy and the safety are poor.
The currently reported synthetic routes of cholesterol have the defects of complicated operation, high pollution, expensive price of used catalysts or problems in some operation steps, and the like, so the reported synthetic routes are not suitable for industrial production. Meanwhile, due to the occurrence of diseases such as mad cow disease, streptococcus suis infection and the like, people pay more and more attention to the safe production of cholesterol, so that the research and development of the efficient, green and economic cholesterol synthesis method based on plant source raw materials has important significance and industrial value.
Disclosure of Invention
In order to solve the defects in the prior art, the invention aims to provide a method for synthesizing cholesterol by taking BA as a raw material. The method takes a plant source raw material 21-hydroxy-20-methylpregn-4-en-3-one ((20S) -21-hydroxy-20-methylpregn-4-en-3-one), also called di-carbinol or BA (bisnalcohol), as a raw material, and synthesizes the cholesterol through the steps of oxidation, Wittig reaction, acetylation, reduction, selective hydrogenation reduction and the like, wherein the purity can reach the standard of commercial grade cholesterol (> 95%). The raw materials for synthesizing the cholesterol are safe and economic, the synthesis method is simple to operate, the yield is high, the total yield can reach 78.5 percent, the cost is low, and the industrial production is convenient.
The raw material BA (bisnalhol) used by the invention is derived from the fermentation of the waste plant sterol of the oil and fat process, is a green raw material of plant source, has annual output reaching kiloton level at present, has low price, and can well avoid the risk of pathogenic bacteria and virus infection possibly existing in the animal source raw material in the prior art.
In the present synthesis method, the raw material BA includes, but is not limited to, those obtained by biological fermentation of phytosterol, or those obtained by chemical synthesis.
The invention provides a method for synthesizing cholesterol by using BA raw material, which comprises the following steps:
step (a), in a first solvent, performing oxidation reaction on BA shown in a formula (1) to obtain a compound shown in a formula (2);
in a second solvent, carrying out a Wittig reaction on the compound shown in the formula (2) to obtain a compound shown in the formula (3);
in a third solvent, performing acetylation reaction on the compound in the formula (3) to obtain a compound in a formula (4);
in a fourth solvent, carrying out reduction reaction on the compound shown in the formula (4) to obtain compounds shown in the formula (5) and the formula (6);
in a fifth solvent, carrying out selective hydrogenation reduction reaction on the compound shown in the formula (5) to obtain the cholesterol; wherein the reaction process of the method is shown as a scheme (A):
in step (a) of the present invention, the oxidation reaction specifically comprises: in a first solvent, BA shown in a formula (1) is subjected to oxidation reaction with TEMPO, sodium bicarbonate, tetrabutylammonium bromide and an oxidant to obtain a compound shown in a formula (2).
Wherein the molar ratio of BA, TEMPO, sodium bicarbonate, tetrabutylammonium bromide and an oxidant shown in the formula (1) is 1: (0.01-1): (1.35-20): (0.1-1): (1.15-5); preferably, 1: 0.01: 1.35: 0.1: 1.15.
wherein the oxidation reaction is carried out under the action of an oxidant, and the oxidant is selected from one or more of N-chlorosuccinimide NCS, N-bromosuccinimide NBS, 2-iodosylbenzoic acid IBX and the like; preferably, it is N-chlorosuccinimide NCS.
Wherein the first solvent is selected from one or more of dichloromethane, tetrahydrofuran, toluene, dimethyl sulfoxide, water and the like; preferably, the solvent is a mixed solvent of dichloromethane and water (volume ratio V/V-5/2).
Wherein the temperature of the oxidation reaction is 0-30 ℃; preferably, it is 0 ℃.
Wherein the time of the oxidation reaction is 3-8 h; preferably, it is 6 h.
In one embodiment, the step of synthesizing the compound of formula (2) comprises: dissolving BA shown in the formula (1) in a first solvent, adding TEMPO, sodium bicarbonate, tetrabutylammonium bromide and NCS, and carrying out oxidation reaction to obtain a compound shown in the formula (2).
In the step (b), the Wittig reaction is specifically as follows: in a second solvent, the compound of the formula (2), 3-dimethylallyl halide, triphenylphosphine and potassium tert-butoxide are subjected to a Wittig reaction to obtain the compound of the formula (3).
Wherein the mol ratio of the compound shown in the formula (2), the 3, 3-dimethylallyl halide, the triphenylphosphine and the potassium tert-butoxide is 1: (1.5-4): (1.5-4): (1-4); preferably, 1: 2: 2: 1.8 or 1: 4: 4: 3.6.
wherein the second solvent is one or more of toluene, benzene, tetrahydrofuran, n-hexane and the like; preferably, it is toluene.
Wherein the temperature of the Wittig reaction is 60-140 ℃; preferably, it is 135 ℃.
Wherein the Wittig reaction time is 4-9 h; preferably, it is 5h or 8 h.
In the step (c), the acetylation reaction is specifically: and (3) performing acetylation reaction on the compound in the formula (3), acetyl chloride, acetic anhydride and pyridine in a third solvent to obtain the compound in the formula (4).
Wherein, the mol ratio of the compound shown in the formula (3), acetyl chloride, acetic anhydride and pyridine is as follows: 1: (25-62.5): (25-62.5): (4-6); preferably, 1: 25: 25: 5.
wherein the third solvent is one or more of acetic anhydride, acetyl chloride, ethyl acetate, dichloromethane and the like; preferably, it is a mixed solvent of acetyl chloride and acetic anhydride (molar ratio of 1: 1).
Wherein the temperature of the acetylation reaction is 40-110 ℃; preferably, it is 100 ℃.
Wherein the acetylation reaction time is 3-5 h; preferably, it is 3.5 h.
In the acetylation reaction, acetyl chloride and acetic anhydride are used as reactants and solvents.
In one embodiment, the step of synthesizing the compound of formula (4) comprises: adding acetyl chloride, acetic anhydride and pyridine into the compound of the formula (3) to perform acetylation reaction to obtain a compound of a formula (4).
In the step (d), the reduction reaction is specifically: and (3) carrying out reduction reaction on the compound of the formula (4) and a reducing agent in a fourth solvent to obtain compounds of the formula (5) and the formula (6).
Wherein the molar ratio of the compound shown in the formula (4) to the reducing agent is 1 (15-25); preferably, 1: 15.
wherein the fourth solvent is one or more of tetrahydrofuran, ethanol, water, dichloromethane, 2-methyltetrahydrofuran, isopropanol, acetic acid, methyl tert-butyl ether, etc.; preferably, the solvent is a mixed solvent of tetrahydrofuran, ethanol and water (volume ratio V/V: 16/8/5).
Wherein the reducing agent is NaBH4、KBH4One or more of the following; preferably, it is NaBH4。
Wherein the temperature of the reduction reaction is 0-30 ℃; preferably, it is 25 ℃.
Wherein the time of the reduction reaction is 6-9 h; preferably, it is 8 h.
In one embodiment, the step of synthesizing the compounds of formula (5) and formula (6) comprises: dissolving the compound of the formula (4) in a fourth solvent, and carrying out reduction reaction with a reducing agent to obtain the compounds of the formula (5) and the formula (6).
In the step (e), the selective hydrogenation reduction reaction specifically comprises: and (3) carrying out selective hydrogenation reduction reaction on the compound shown in the formula (5) and a reducing agent in the fifth solvent under the action of a catalyst to obtain the cholesterol.
Wherein, the method also comprises a purification step before obtaining the cholesterol, and the purification step is one or more of column chromatography, recrystallization, pulping and the like.
Wherein the catalyst is RaneyNi.
Wherein the reducing agent is H2。
Wherein the mass ratio of the compound shown in the formula (5) to the catalyst RaneyNi is 1: (0.05-5); preferably, 1: 2.
wherein the fifth solvent is selected from one or more of isopropanol, dichloromethane, methanol, 2-methyltetrahydrofuran, tetrahydrofuran, ethanol, water, methyl tert-butyl ether, ethyl acetate, toluene and the like; preferably, it is isopropanol.
Wherein the temperature of the hydrogenation reduction reaction is 0-60 ℃; preferably, it is 30 ℃.
Wherein, the reducing agent H of the hydrogenation reduction reaction2The pressure is 1-20atm, preferably 1 atm.
Wherein the time of the hydrogenation reduction reaction is 6-10 h; preferably, it is 7 h.
In one embodiment, the step of synthesizing cholesterol comprises: dissolving the compound of formula (5) in a fifth solvent, adding Raney Ni and H2After the displacement, selective hydrogenation reduction reaction is carried out to obtain the cholesterol.
The invention also provides a compound, and the structure of the compound is shown as the formula (4):
in the compounds of the above formula (3), formula (4), formula (5) and formula (6) of the present invention, since the double bond of the side chain of the D ring introduced by the Wittig reaction has a cis-trans configuration, the compounds of the formula (3), formula (4), formula (5) and formula (6) are not single substances, and are all mixtures.
The compound shown in the formula (6) is catalyzed by Raney Ni and is selectively hydrogenated and reduced to obtain the surface cholesterol.
The beneficial effects of the invention include: according to the preparation method of the cholesterol, the used raw material BA is a plant source raw material, so that the risk of pathogenic bacteria and virus infection possibly existing in animal source raw materials is avoided, and the cholesterol is cheap and easy to obtain; the synthesis steps of the cholesterol are simple and convenient, the yield is high, the side reaction is less, the environment is protected, and the industrial production is convenient to realize; solves the safety problem of the prior cholesterol product and the problems of high cost, unfriendly environment and unsuitability for large-scale industrial production in the synthesis technology.
In the Wittig reaction, 3-dimethylallyl bromide is used for synthesizing the Wittig reagent (3, 3-dimethylallyl triphenylphosphine bromide), side reactions are few, impurities are easy to remove, and the reaction yield is improved. In the acetylation reaction, acetyl chloride and acetic anhydride are used as reactants and solvents, so that impurities are prevented from being generated, and the yield is further improved. In the reduction reaction, through screening of a reaction solvent, when a mixture of tetrahydrofuran, ethanol and water is selected as the solvent, the generation of byproducts can be effectively reduced, and the reaction yield is greatly improved. In the selective hydrogenation reduction reaction of the present invention, H is selected2When Raney Ni is used as a catalyst as a reducing agent, the yield of the reaction can be effectively improved when isopropanol is selected as a solvent for the selective hydrogenation reduction reaction through the screening of a reaction solvent.
Drawings
FIG. 1 shows TLC thin plate chromatography of a reaction solution in comparative example I of the present invention.
FIG. 2 shows TLC plate chromatography of the reaction solution in comparative example I of the present invention.
FIG. 3 shows TLC plate chromatography of the reaction solution in comparative example II of the present invention.
FIG. 4 shows TLC plate chromatography of the reaction solution in comparative example II of the present invention.
FIG. 5 shows TLC thin plate chromatography of the reaction solution in comparative example III of the present invention.
FIG. 6 shows TLC plate chromatography of the reaction solution in comparative example four of the present invention.
Fig. 5, TLC: PE/EA is 20: 1; in addition to fig. 5, TLC: PE/EA is 5: 1.
FIG. 7 is a gas chromatogram of the crude cholesterol prepared by the method (1) in the fifth embodiment of the present invention.
FIG. 8 is a gas chromatogram of the fine cholesterol prepared by the method (1) in the fifth embodiment of the present invention.
FIG. 9 is a gas chromatogram of the crude cholesterol prepared by the method (2) in the fifth embodiment of the present invention.
FIG. 10 is a gas chromatogram of the fine cholesterol prepared by the method (2) in the fifth embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the following specific examples and the accompanying drawings. The procedures, conditions, experimental methods and the like for carrying out the present invention are general knowledge and common general knowledge in the art except for the contents specifically mentioned below, and the present invention is not particularly limited.
In the following examples, the structures of the compounds were determined using a nuclear magnetic resonance apparatus and a high-resolution mass spectrometer; the reagent is mainly provided by Shanghai national drug chemical reagent company; the product is purified mainly by pulping and column chromatography; silica gel (200- & 300) was produced by Qingdao maritime works.
EXAMPLES preparation of Compounds of formula (2)
A1000 mL single-neck flask was charged with BA (50.00g, 0.15mol), TEMPO (235mg, 1.50mmol), dichloromethane (400mL), sodium bicarbonate (17.60g, 0.21mol), NCS (23.10g, 173.00mmol), tetrabutylammonium bromide (4.84g, 15mmol) and water (160mL) in that order and reacted at 0 ℃ for 6 h. After completion of TLC detection reaction, sodium thiosulfate pentahydrate solution (11.25g sodium thiosulfate pentahydrate/225 mL water) was added, stirred at 5-10 deg.C for 20min, separated, the aqueous phase was extracted with dichloromethane (100 mL. times.3), the organic layers were combined, washed with 1% sodium hydroxide solution (300mL), separated, the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the compound of formula (2) (47.21g, white solid, 95% molar yield).1H NMR(400MHz,CDCl3)δ9.55(s,1H),5.71(s,1H),2.45-2.23(m,5H),1.99(t,J＝13.7Hz,2H),1.91-1.78(m,2H),1.68(t,J＝10.2Hz,2H),1.43(m,5H),1.30-1.19(m,2H),1.17(s,3H),1.11(d,J＝5.5Hz,3H),1.06-0.89(m,3H),0.75(s,3H).13C NMR(100MHz,CDCl3)δ205.00,199.65,171.31,123.99,55.25,53.84,51.04,49.54,43.10,39.39,38.68,35.80,35.68,34.06,32.93,32.05,27.11,24.64,21.06,17.48,13.53,12.44.HRMS(ESI):calcd forC22H32NaO2[M+Na]+,351.2295,found 351.2292.
EXAMPLE two preparation of Compound (3)
This example shows the results of the preparation of a compound of formula (3) under 4 different experimental conditions:
(1) triphenylphosphine (23.97g, 91.40mmol), 3-dimethylallyl bromide (13.62g, 91.40mmol) and 230mL of toluene were put into a 500mL single-neck flask, and after refluxing at 135 ℃ for 2 hours, the flask was cooled to room temperature, potassium tert-butoxide (9.23g, 82.26mmol) was added under ice bath, and after stirring for 0.5 hour, the compound of formula (2) (15.00g, 45.70mmol) was added, and the mixture was heated to 135 ℃ and refluxed for 2.5 hours. After the completion of the reaction monitored by TLC, the reaction was cooled to room temperature, the solvent was evaporated, dichloromethane was added, 2M HCl (12mL) was added to adjust the PH to neutral, the organic phase was washed with water, a saturated aqueous NaCl solution (100mL × 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure and slurried with a mixed solution of methyl tert-butyl ether and methanol (30mL, V/V ═ 5/1) to give the compound of formula (3) (3E and 3Z, 3E/3Z ≈ 87/13, 16.72g of a white solid, 96% molar yield).
Note: in the invention, the proportion of the E/Z configuration of the intermediate compounds 4, 5 and 6 obtained by acetylating and reducing the compound 3 is basically kept unchanged, the double bond at the C-22 position is mainly the E configuration, and the Z configuration is auxiliary (3E/3Z is approximately equal to 87/13). In addition, cholesterol is obtained by hydrogenation reduction of cis-trans isomers of C-22 double bond in the compound 5 through Raney nickel, so that the proportion of E/Z configuration of the corresponding compound is not indicated in the following examples.
(2) Triphenylphosphine (47.95g, 182.80mmol), 3-dimethylallyl chloride (19.12g, 182.80mmol) and 230mL of toluene were placed in a 500mL single-neck flask, and reacted at 135 ℃ for 4 hours under reflux, followed by cooling to room temperature, potassium tert-butoxide (18.46g, 164.52mmol) was added under ice-cooling, followed by stirring for 0.5 hours, followed by addition of the compound of formula (2) (15.00g, 45.70mmol), and then heated to 135 ℃ for 4.5 hours under reflux. After completion of the reaction monitored by TLC, the reaction was cooled to room temperature, the solvent was evaporated, dichloromethane was added, 2M HCl (12mL) was added to adjust the PH to neutral, the organic phase was washed with water, a saturated aqueous NaCl solution (100mL × 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure and slurried with a mixed solution of methyl tert-butyl ether and methanol (30mL, V/V ═ 5/1) to give compounds of formula (3) (3E and 3Z, 12.51g of white solids, 72% molar yield).
(3) Triphenylphosphine (13.19g, 50.27mmol), 3-dimethylallyl bromide (7.49g, 50.27mmol) and 230mL of toluene were put into a 500mL single-neck flask, and after refluxing at 135 ℃ for 2 hours, the flask was cooled to room temperature, potassium tert-butoxide (9.23g, 82.26mmol) was added under ice bath, and after stirring for 0.5 hour, the compound of formula (2) (15.00g, 45.70mmol) was added, and the mixture was heated to 135 ℃ and refluxed for 2.5 hours. After completion of the reaction monitored by TLC, the reaction was cooled to room temperature, the solvent was evaporated, dichloromethane was added, 2M HCl (12mL) was added to adjust the PH to neutral, the organic phase was washed with water, a saturated aqueous NaCl solution (100mL × 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure and slurried with a mixed solution of methyl tert-butyl ether and methanol (30mL, V/V ═ 5/1) to give compounds of formula (3) (3E and 3Z, 13.90g of white solids, 80% molar yield).
(4) Triphenylphosphine (17.62g, 67.18mmol), 3-dimethylallyl bromide (10.01g, 67.18mmol) and 230mL of toluene were placed in a 500mL single-neck flask, and after refluxing at 135 ℃ for 2 hours, the flask was cooled to room temperature, potassium tert-butoxide (9.23g, 82.26mmol) was added under ice-cooling, and after stirring for 0.5 hour, the compound of formula (2) (15.00g, 45.70mmol) was added, and the mixture was heated to 135 ℃ and refluxed for 2.5 hours. After completion of the reaction monitored by TLC, the reaction was cooled to room temperature, the solvent was evaporated, dichloromethane was added, 2M HCl (12mL) was added to adjust the PH to neutral, the organic phase was washed with water, a saturated aqueous NaCl solution (100mL × 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure and slurried with a mixed solution of methyl tert-butyl ether and methanol (30mL, V/V ═ 5/1) to give compounds of formula (3) (3E and 3Z, 14.77g of white solids, 85% molar yield).1H NMR(500MHz,CDCl3)δ6.17-6.08(m,1H),5.77-5.69(m,2H),5.40-5.35(m,1H),2.45-2.22(m,6H),2.13-2.07(m,1H),2.04-1.96(m,1H),1.92-1.79(m,4H),1.76-1.72(m,8H),1.70-1.64(m,2H),1.62(d,J＝0.9Hz,3H),1.54-1.39(m,3H),1.38-1.17(m,5H),1.15(s,4H),1.07-0.96(m,5H),0.92(d,J＝6.6Hz,3H),0.68(s,3H).13C NMR(125MHz,CDCl3)δ199.74,171.79,138.55,132.59,125.25,124.27,123.73,56.32,55.69,53.88,42.51,40.63,38.84,38.65,35.68,35.60,34.01,32.97,32.06,27.87,25.96,24.08,21.50,20.88,18.27,17.39,12.24.HRMS(ESI):calcd for C27H40NaO[M+Na]+,403.2971,found 403.2967.
EXAMPLE III preparation of Compound of formula (4)
This example shows the results of the preparation of a compound of formula (4) under 5 different experimental conditions:
(1) a 50mL single-neck flask was charged with the compound of formula (3) (3E and 3Z, 500mg, 1.31mmol), acetic anhydride (3.34g, 32.75mmol), acetyl chloride (2.57g, 32.75mmol) and pyridine (518mg, 6.55mmol), refluxed at 100 ℃ for 3.5h, cooled to room temperature after TLC monitoring completion of the reaction, evaporated to remove acetyl chloride and acetic anhydride, added dichloromethane (150mL), washed with organic phase water (80mL × 3), separated the organic phase, dried over anhydrous sodium sulfate, evaporated to remove the solvent, and purified by column chromatography (petroleum ether/ethyl acetate: 80/1, v/v) to give the compound of formula (4) (4E and 4Z, 528mg of white solid, 95% molar yield).
(2) A 50mL single-neck flask was charged with the compound of formula (3) (3E and 3Z, 500mg, 1.31mmol), acetic anhydride (3.34g, 32.75mmol), acetyl chloride (2.57g, 32.75mmol) and pyridine (621mg, 7.86mmol), refluxed at 110 ℃ for 4h, TLC monitored for completion of the reaction, cooled to room temperature, evaporated to remove acetyl chloride and acetic anhydride, added dichloromethane (150mL), washed with organic phase water (80mL × 3), separated from the organic phase, dried over anhydrous sodium sulfate, evaporated to remove the solvent, and purified by column chromatography (petroleum ether/ethyl acetate ═ 80/1, v/v) to give the compound of formula (4) (4E and 4Z, 516mg of white solid, 93% molar yield).
(3) A 50mL single-neck flask was charged with the compound of formula (3) (3E and 3Z, 500mg, 1.31mmol), acetic anhydride (6.02g, 58.95mmol), acetyl chloride (2.57g, 32.75mmol) and pyridine (518mg, 6.55mmol), refluxed at 90 ℃ for 4h, TLC monitored for completion of the reaction, cooled to room temperature, evaporated to remove acetyl chloride and acetic anhydride, added dichloromethane (150mL), washed with an organic phase (80mL × 3), separated from the organic phase, dried over anhydrous sodium sulfate, evaporated to remove the solvent, and purified by column chromatography (petroleum ether/ethyl acetate ═ 80/1, v/v) to give the compound of formula (4) (4E and 4Z, 505mg of white solid, 91% molar yield).
(4) A 50mL single-neck flask was charged with the compound of formula (3) (3E and 3Z, 500mg, 1.31mmol), acetic anhydride (6.02g, 58.95mmol), acetyl chloride (2.57g, 32.75mmol) and pyridine (621mg, 7.86mmol), refluxed at 100 ℃ for 4h, TLC monitored for completion of the reaction, cooled to room temperature, evaporated to remove acetyl chloride and acetic anhydride, added dichloromethane (150mL), washed with an organic phase (80mL × 3), separated from the organic phase, dried over anhydrous sodium sulfate, evaporated to remove the solvent, and purified by column chromatography (petroleum ether/ethyl acetate ═ 80/1, v/v) to give the compound of formula (4) (4E and 4Z, 496mg of white solid, 89% molar yield).
(5) In a 50mL single-neck flask were charged the compound of formula (3) (3E and 3Z, 500mg, 1.31mmol), acetic anhydride (3.21g, 31.44mmol), acetyl chloride (2.47g, 31.44mmol), refluxed at 65 ℃ for 4 hours, TLC monitored the completion of the reaction, cooled to room temperature, evaporated to remove acetyl chloride and acetic anhydride, added dichloromethane (150mL), organic phase washed with water (80mL × 3), organic phase separated, dried over anhydrous sodium sulfate, evaporated to remove solvent, purified by column chromatography (petroleum ether/ethyl acetate ═ 80/1, v/v) to give the compound of formula (4) (4E and 4Z, white solid 461mg, molar yield 83%).1H NMR(500MHz,CDCl3)δ6.18-6.13(m,1H),5.78(d,J＝10.8Hz,1H),5.71(d,J＝2.2Hz,1H),5.46-5.39(m,2H),2.50-2.41(m,1H),2.15(s,5H),2.05-2.01(m,1H),1.90-1.82(m,1H),1.78-1.75(m,6H),1.72-1.63(m,3H),1.51-1.40(m,1H),1.37-1.16(m,5H),1.07(d,J＝6.6Hz,3H),1.60-1.56(m,8H),1.03(d,J＝3.5Hz,4H),0.75(s,3H).13C NMR(125MHz,CDCl3)δ169.43,146.98,139.38,138.53,132.65,125.28,124.11,124.08,117.02,56.86,55.91,48.00,42.48,40.27,39.64,34.92,33.79,31.85,31.76,28.56,25.92,24.82,24.18,21.21,21.11,20.64,18.86,18.23,12.20.HRMS(ESI):calcd for C29H42NaO2[M+Na]+,445.3077,found 445.3081.
EXAMPLES preparation of Compounds of formulae (5) and (6)
This example gives the results of the preparation of compounds of formulae (5) and (6) under 5 different experimental conditions:
(1) in a 100mL single-neck flask were added the compound of formula (4) (4E and 4Z, 486mg, 1.15mmol), a mixed solvent of tetrahydrofuran, ethanol and water (23.5mL, V/V ═ 16/8/5), and sodium borohydride (653mg, 17.25mmol) was added at 0 ℃ to react for 8h at 25 ℃. After the completion of the reaction was monitored by TLC, 2m naoh solution (10mL) was added, the solvent was evaporated, dichloromethane (50mL × 3) was added for extraction, the organic phase was washed with water, a saturated NaCl solution (50mL × 3) was washed, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (petroleum ether/ethyl acetate ═ 40/1, v/v) to obtain compounds of formula (5) (5E and 5Z, 403mg of white solid, molar yield 92%) and compounds of formula (6) (6E and 6Z, 30mg of white solid, molar yield 7%).
(2) In a 100mL single-neck flask were added the compound of formula (4) (4E and 4Z, 523mg, 1.24mmol), a mixed solvent of tetrahydrofuran, ethanol and water (20.1mL, V/V ═ 16/4/3), sodium borohydride (938mg, 24.80mmol) was added at 0 ℃, and reaction was carried out for 8h at 25 ℃. After the completion of the reaction was monitored by TLC, 2m naoh solution (10mL) was added, the solvent was evaporated, dichloromethane (50mL × 3) was added for extraction, the organic phase was washed with water, a saturated NaCl solution (50mL × 3) was washed, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (petroleum ether/ethyl acetate ═ 40/1, v/v) to obtain compounds of formula (5) (5E and 5Z, 415mg of white solid, 88% molar yield) and compounds of formula (6) (6E and 6Z, 47mg of white solid, 10% molar yield).
(3) In a 100mL single-neck flask were added the compound of formula (4) (4E and 4Z, 500mg, 1.18mmol), a mixed solvent of tetrahydrofuran, ethanol and water (22.5mL, V/V ═ 16/8/3), sodium borohydride (1.12g, 29.50mmol) was added at 0 ℃, and reaction was carried out for 8h at 25 ℃. After the completion of the reaction monitored by TLC, 2m naoh solution (10mL) was added, the solvent was distilled off, dichloromethane (50mL × 3) was added for extraction, the organic phase was washed with water, a saturated NaCl solution (50mL × 3) was washed, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (petroleum ether/ethyl acetate ═ 40/1, v/v) to obtain compounds of formula (5) (5E and 5Z, 394mg of white solid, molar yield 87%) and compounds of formula (6) (6E and 6Z, 54mg of white solid, molar yield 12%).
(4) A 100mL single-neck flask was charged with the compound of formula (4) (4E and 4Z, 500mg, 1.18mmol), a mixed solvent of methanol and dichloromethane (6mL, V/V ═ 2/1), and sodium borohydride (893mg, 23.6mmol) was added at 0 ℃ to react at 25 ℃ for 8 hours. After the completion of the reaction was monitored by TLC, 2M NaOH solution (10mL) was added, the solvent was evaporated, dichloromethane (50mL × 3) was added for extraction, the organic phase was washed with water, a saturated NaCl solution (50mL × 3) was washed, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (petroleum ether/ethyl acetate ═ 40/1, v/v) to obtain compounds of formula (5) (5E and 5Z, white solid 353mg, molar yield 78%) and compounds of formula (6) (6E and 6Z, white solid 91mg, molar yield 20%).
(5) In a 100mL single-neck flask, the compound of formula (4) (4E and 4Z, 420mg, 0.99mmol), a mixed solvent of tetrahydrofuran, ethanol and water (20.3mL, V/V ═ 16/8/5), potassium borohydride (801mg, 14.85mmol) was added at 0 ℃, and the reaction was carried out at 25 ℃ for 8 hours. After the completion of the reaction was monitored by TLC, 2m naoh solution (10mL) was added, the solvent was evaporated, dichloromethane (50mL × 3) was added for extraction, the organic phase was washed with water, a saturated NaCl solution (50mL × 3) was washed, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (petroleum ether/ethyl acetate ═ 40/1, v/v) to obtain compounds of formula (5) (5E and 5Z, 120mg of white solid, 32% molar yield) and compounds of formula (6) (6E and 6Z, 75mg of white solid, 20% molar yield).
Note: the reducibility of potassium borohydride is weaker than that of sodium borohydride, the selectivity is poor, side reactions are more, and a 3-position generates more alpha-OH byproducts, so the experimental yield is low.
A compound of formula (5):1H NMR(600MHz,CDCl3)δ6.15-6.11(m,1H),5.75(d,J＝10.6Hz,1H),5.43-5.31(m,2H),3.55-3.47(m,1H),2.33-2.19(m,2H),2.14-2.09(m,1H),2.02-1.92(m,2H),1.86-1.79(m,2H),1.73(d,J＝9.5Hz,6H),1.70-1.64(m,1H),1.57-1.41(m,6H),1.29-1.11(m,3H),1.10-1.05(m,2H),1.04(d,J＝6.6Hz,3H),1.02-0.98(m,4H),0.95-0.89(m,1H),0.70(s,3H).13C NMR(150MHz,CDCl3)δ140.76,138.55,138.55,136.35,132.60,125.30,124.09,121.70,121.69,120.62,71.77,56.79,55.93,50.15,42.37,42.30,40.27,39.68,37.28,36.52,31.91,31.89,31.66,28.56,25.94,24.29,21.09,20.66,19.42,18.25,12.10.HRMS(ESI):calcd for C27H42NaO[M+Na]+,405.3128,found 405.3125.
a compound of formula (6):1H NMR(600MHz,CDCl3)δ6.13(dd,J＝15.0,10.8Hz,1H),5.78-5.72(m,1H),5.43-5.36(m,2H),4.01(t,J＝3.0Hz,1H),2.57(d,J＝14.7Hz,1H),2.15-2.09(m,1H),2.09-2.04(m,1H),2.03-1.94(m,2H),1.74(dd,J＝9.5,1.3Hz,6H),1.71-1.65(m,3H),1.64-1.60(m,1H),1.60-1.51(m,4H),1.50-1.38(m,4H),1.29-1.21(m,3H),1.20-1.15(m,2H),1.05(d,J＝6.8Hz,3H),1.01(s,3H),0.71(s,3H).13C NMR(150MHz,CDCl3)δ138.72,138.68,132.76,125.43,124.22,124.15,67.25,56.90,56.03,50.52,42.49,40.42,40.02,39.79,37.50,33.38,32.10,31.99,29.06,28.68,26.07,24.40,20.93,20.79,18.81,18.37,12.23.HRMS(ESI):calcd for C27H42NaO[M+Na]+,405.3128,found 405.3130.
EXAMPLE preparation of five Cholesterol
This example shows the results of cholesterol preparation under 2 different experimental conditions:
(1) a100 mL single neck flask was charged with compound of formula (5) (5E and 5Z, 1.0g, 2.62mmol), Raney Ni (2.0g, wet weight), isopropanol (30mL), H2(1atm), reaction at 30 ℃ for 7h, TLC monitoring the completion of the reaction, filtering to remove raney ni, concentrating the filtrate under reduced pressure to give crude cholesterol (gas chromatography purity 93.28%, fig. 7), purifying by column chromatography (petroleum ether/ethyl acetate 5/1, v/v) to give cholesterol (995 mg white solid, 98.5% molar yield,gas chromatography purity 95.87%, fig. 8).
(2) A100 mL single-neck flask was charged with compound of formula (5) (5E and 5Z, 2.0g, 5.23mmol), Raney Ni (4.0g, wet weight), a mixed solvent of dichloromethane and methanol (50mL, V/V ═ 1/4), H2(1atm), reaction at 30 ℃ for 7h, TLC monitoring the completion of the reaction, filtration to remove raney ni, concentration of the filtrate under reduced pressure to give crude cholesterol (gas chromatography purity 93.26%, fig. 9), purification by recrystallization (absolute ethanol/water 9/1, v/v) to give cholesterol (1.72 g white solid, 86% molar yield, 95.09% gas chromatography purity, fig. 10). mp: 147-.1H NMR(600MHz,CDCl3)δ5.36-5.34(m,1H),3.55-3.49(m,1H),2.35-2.19(m,2H),2.06-1.92(m,2H),1.86-1.79(m,3H),1.68-1.19(m,14H),1.19-1.03(m,7H),1.01(s,3H),0.98-0.93(m,1H),0.91(d,J＝6.6Hz,3H),0.86(dd,J＝6.6,2.8Hz,6H),0.68(s,3H).13C NMR(150MHz,CDCl3)δ140.77,121.73,71.82,56.78,56.17,50.14,42.33,42.32,39.80,39.53,37.27,36.52,36.20,35.80,31.93,31.92,31.68,28.25,28.03,24.31,23.84,22.84,22.58,21.10,19.41,18.73,11.87.HRMS(ESI):calcd for C27H46NaO[M+Na]+,409.3441,found 409.3121.
Impurity structures which may be introduced during the process of obtaining cholesterol from compound 5(5E and 5Z) by Raney Ni hydrogenation reduction are as follows, such as compound 7-1, 7-2, 7-3, 7-4:
in example five, in the gas chromatogram of the crude cholesterol obtained in experimental method (1) (fig. 7), the peak having a retention time of 7.987min (93.28%) was the peak of cholesterol; the retention time is 7.200min (0.6%), 7.374min (1.52%), 7.723min (1.01%) and the corresponding compound is one of the partially reduced impurities 7-1, 7-2, 7-3; the compound corresponding to the impurity peak with a retention time of 8.096min (3.11%) was the over-reduced impurity 7-4.
In example five, in the gas chromatogram of the purified fine cholesterol obtained by the experimental method (1) column chromatography (fig. 8), the peak with the retention time of 7.992min (95.87%) was the peak of cholesterol; the retention time is 7.193min (0.58%), 7.364min (2.01%), 7.705min (0.08%), the compound corresponding to each of the three impurity peaks is one of the partially reduced impurities 7-1, 7-2, 7-3; the compound corresponding to the impurity peak having a retention time of 8.087min (0.91%) was the over-reduced impurity 7-4.
In example five, in the gas chromatogram of the crude cholesterol obtained in experimental method (2) (fig. 9), the peak having a retention time of 8.073min (93.26%) was the peak of cholesterol; the retention time is 7.296min (0.26%), 7.464min (2.52%), 7.596min (0.17%), the corresponding compound is one of the partially reduced impurities 7-1, 7-2, 7-3; the compound corresponding to the impurity peak with a retention time of 8.192min (3.36%) was the over-reduced impurity 7-4.
In example five, in the gas chromatogram of the purified fine cholesterol obtained by the recrystallization purification of experimental method (2) (fig. 10), the peak having a retention time of 8.026min (95.09%) was the peak of cholesterol; the retention time is 7.223min (0.57%), 7.405min (2.17%), 7.755min (0.12%), the compound corresponding to each of the three impurity peaks is one of the partially reduced impurities 7-1, 7-2, 7-3; the compound corresponding to the impurity peak with a retention time of 8.122min (1.95%) was the over-reduced impurity 7-4.
Comparative example 1
In patent literature (background art Scheme 2, CN105218610A), cholesterol was synthesized in a total yield of 67% by 5-step reaction using a degraded stigmasterol as a raw material. The first reaction step in the technical route of this patent document is shown in the following reaction formula one:
patent document CN105218610A describes a reaction formula in which compound 02 obtained by oxidizing BA is used as a raw material, ethanol is used as a solvent, and the reaction is carried out under the action of p-toluenesulfonic acid and triethyl orthoformate by heating to 40 ℃ and keeping the temperature for 4 hours to obtain compound 03. Molar yield: 97.50 percent.
According to the experimental method provided by the patent document, the ethanol is used as a solvent, the compound 02 is used as a substrate, the reaction is carried out for 4 hours at 40 ℃ under the catalysis of p-toluenesulfonic acid and triethyl orthoformate, the TLC detects that the raw materials are completely reacted (as shown in figure 1), and the compound 03' (shown in a reaction formula II) is obtained by post-treatment according to the method of the patent document (CN105218610A), which is different from the compound described in the patent document. In the present invention, it was also attempted to reduce the amount of triethyl orthoformate, and the TLC detection showed that the starting material reaction was complete (as shown in fig. 2), but the result of the reaction formula one as described in patent document (CN105218610A) was not obtained, but the result of the reaction formula three as shown below was obtained. It is described that when the carbonyl group at the 3-position of compound 02 is protected according to the method reported in patent document CN105218610A, the aldehyde group at the C-22 position is preferentially protected to form acetal, and compounds 03 'and 03' represented by the reaction formula II or III are formed, and the compound 03 described in patent document (CN105218610A) cannot be obtained, and it is obvious that the compounds 03 'and 03' represented by the reaction formula II or III cannot undergo the subsequent Wittig reaction.
The experimental method comprises the following steps: in a 50mL single neck flask was added 4mL of ethanol, 2mL of triethyl orthoformate, the compound of formula (02) (2.00g, 70.20mmol), p-toluenesulfonic acid (20mg, 0.12mmol), and the reaction was incubated at 40 ℃ for 4h with TLC to detect that the starting material had reacted to completion (as shown in FIG. 1). Sodium acetate (20mg) was added in an ice bath, water was added, the cake was washed with water until the eluate was neutral, and after draining, column chromatography purification was performed (petroleum ether: ethyl acetate ═ 20:1) to obtain compound 03' (colorless oil 2.30g, molar yield 88%).1H NMR(600MHz,CDCl3)δ5.23-5.19(m1H),5.10(d,J＝1.9Hz,1H),4.30(d,J＝2.4Hz,1H),3.81-3.70(m,4H),3.61-3.55(m,1H),3.49-3.41(m,2H),2.31-2.25(m,1H),2.18-1.96(m,4H),1.85-1.78(m,2H),1.71-1.52(m,7H),1.43-1.33(m,4H),1.29(d,J＝7.0Hz,3H),1.22-1.18(m,6H),0.99(d,J＝6.7Hz,3H),0.96(s,3H),0.70(s,3H).13C NMR(150MHz,CDCl3)δ154.52,141.06,118.03,106.07,99.04,64.60,63.35,62.14,56.48,51.88,48.33,42.59,40.39,39.62,35.18,33.86,31.92,31.86,27.72,25.55,24.42,21.17,18.96,15.49,15.37,14.68,11.94,11.89.HRMS(ESI):calcd for C28H46NaO3[M+Na]+,453.3339,found 453.3328.
The experimental method comprises the following steps: a50 mL single neck flask was charged with 4mL of ethanol, 1mL of triethyl orthoformate, the compound of formula (02) (2.00g, 70.20mmol), p-toluenesulfonic acid (20mg, 0.12mmol), and the reaction was incubated at 40 ℃ for 4h with TLC to detect that the starting material had reacted to completion (as shown in FIG. 2). Sodium acetate (20mg) was added in an ice bath, water was added, the cake was washed with water until the eluate was neutral, and after draining, column chromatography purification was performed (petroleum ether: ethyl acetate 5:1) to obtain compound 03 "(colorless oil 2.32g, molar yield 95%).1H NMR(600MHz,CDCl3)δ5.69(d,J＝1.8Hz,1H),4.27(d,J＝2.4Hz,1H),3.78-3.72(m,1H),3.58-3.53(m,1H),3.47-3.40(m,2H),2.42-2.21(m,4H),2.01-1.97(m,2H),1.84-1.77(m,2H),1.69-1.58(m,3H),1.53-1.47(m,2H),1.41-1.27(m,3H),1.21-1.17(m,6H),1.16(d,J＝4.1Hz,4H),1.13-1.08(m,1H),1.04-0.98(m,2H),0.96(d,J＝6.8Hz,3H),0.93-0.86(m,1H),0.69(s,3H).13C NMR(150MHz,CDCl3)δ199.58,171.53,123.76,105.95,64.58,63.40,55.39,53.81,51.81,42.52,40.33,39.42,35.69,35.63,33.97,32.92,32.03,27.62,24.37,21.01,17.37,15.48,15.36,11.91,11.86.
Comparative example No. two
The second Wittig reaction reported in the patent literature (CN105218610A) is represented by the following equation IV:
in this patent document, triphenylphosphine and 1-chloro-3-methylbutane are added to toluene as a solvent, and refluxed for 2 hours, potassium tert-butoxide and the compound 03 are added, and the reaction is refluxed for 4 hours to obtain the compound 04 in molar yield: 90.29 percent. The Wittig reaction of the compound of formula (2) according to the present invention using 1-chloro-3-methylbutane or 1-bromo-3-methylbutane as in the present invention using the method of patent document (CN105218610A) (comparative example II, reaction formula IV) gave compound 3' as shown in reaction formula V, but did not give the expected compound 3, indicating that the method disclosed in patent document (CN105218610A) could not be applied to the present invention.
The experimental method comprises the following steps: triphenylphosphine (797mg, 3.04mmol), 1-chloro-3-methylbutane (324mg, 3.04mmol) and toluene (10mL) were added to a 50mL single-neck flask, and after refluxing at 135 ℃ for 2 hours, the flask was cooled to room temperature, potassium tert-butoxide (307mg, 2.74.02mmol) was added in 3 portions under ice bath, and after stirring at ice bath for 0.5 hour, the compound of formula (2) (500mg, 1.52mmol) was added, and then refluxing at 135 ℃ for 4 hours. After completion of the reaction was monitored by TLC (as shown in fig. 3), the reaction was cooled to room temperature, the solvent was removed by rotary evaporation, dichloromethane (50mL) was added, 2M HCl (4mL) was added to adjust the solution to neutrality, the organic phase was washed with water, with saturated aqueous NaCl (50mL × 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography (PE: EA ═ 20:1) gave compound 3' (516 mg of colorless oil, 85% molar yield).
1H NMR(600MHz,CDCl3)δ9.53(d,J＝5.0Hz,1H),2.72-2.68(m,1H),2.39-2.25(m,6H),2.10(dd,J＝14.4,4.8Hz,1H),1.94-1.86(m,3H),1.68-1.53(m,8H),1.43-1.30(m,4H),1.25(s,3H),1.15(s,3H),1.04(d,J＝6.7Hz,3H),0.89(d,J＝6.6Hz,6H),0.72(s,3H).13C NMR(150MHz,CDCl3)δ205.71,198.54,163.30,133.32,55.49,54.31,51.82,48.85,42.21,39.05,38.94,38.40,35.35,35.16,33.95,32.12,29.71,28.43,27.31,26.49,23.74,23.21,22.54,20.74,17.80,13.60,12.97.HRMS(ESI):calcdforC27H42NaO2[M+Na]+,421.3077,found 421.3075.
A50 mL single-neck flask was charged with triphenylphosphine (797mg, 3.04mmol), 1-bromo-3-methylbutane (459mg, 3.04mmol), and toluene (10mL), and reacted at 135 ℃ for 2 hours under reflux, followed by cooling to room temperature, addition of potassium tert-butoxide (307mg, 2.74mmol) in portions under ice bath, stirring for 0.5 hours under ice bath, addition of the compound of formula (2) (500mg, 1.52mmol), and reaction at 135 ℃ for 2.5 hours under reflux. After completion of the reaction was monitored by TLC (as shown in fig. 4), the reaction was cooled to room temperature, the solvent was removed by rotary evaporation, dichloromethane (50mL) was added, 2M HCl (4mL) was added to adjust the solution to neutrality, the organic phase was washed with water, with saturated aqueous NaCl (50mL × 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography (PE: EA ═ 20:1) gave compound 3' (534 mg of colorless oil, 88% molar yield).
1H NMR(600MHz,CDCl3)δ9.53(d,J＝5.0Hz,1H),2.72-2.68(m,1H),2.39-2.25(m,6H),2.10(dd,J＝14.4,4.8Hz,1H),1.94-1.86(m,3H),1.68-1.53(m,8H),1.43-1.30(m,4H),1.25(s,3H),1.15(s,3H),1.04(d,J＝6.7Hz,3H),0.89(d,J＝6.6Hz,6H),0.72(s,3H).13C NMR(150MHz,CDCl3)δ205.71,198.54,163.30,133.32,55.49,54.31,51.82,48.85,42.21,39.05,38.94,38.40,35.35,35.16,33.95,32.12,29.71,28.43,27.31,26.49,23.74,23.21,22.54,20.74,17.80,13.60,12.97.HRMS(ESI):calcdforC27H42NaO2[M+Na]+,421.3077,found 421.3075.
Comparative example No. three
The 3-position ester group of the compound of formula 4 of the present invention is NaBH4Reducing to obtain a compound shown in formula 5, and then performing Raney Ni/H treatment on a side chain double bond of the compound shown in formula 52Reducing and synthesizing the cholesterol. Wherein the reducing agent used for reducing the side chain double bond of the compound of the formula 5 is H2The catalyst is Raney Ni shown as the following reaction formula VI:
meanwhile, the invention tries to use Raney Ni/H first2As a reducing agent, the double bond of the side chain is subjected to selective hydrogenation reduction reaction and then NaBH is used4Reducing the ester group at the 3-position. The reaction results are shown in equation seven, and compounds 6 and 6' were obtained (the case of TLC detection of the reaction solution is shown in FIG. 5), while the expected compound 7 was not obtained, indicating Raney Ni/H2As the reducing agent, selective hydrogenation reduction of the side chain cannot be carried out, and therefore, the reduction reaction sequence shown in the reaction formula VICannot be changed.
The experimental method comprises the following steps: a100 mL single neck flask was charged with compound of formula (4) (2.10g, 17.2mmol), Raney Ni (4.20g, wet weight), isopropanol (30mL), H2(1atm), reaction at 30 ℃ for 11h, TLC monitoring completion of the reaction, filtration to remove Raney Ni, concentration of the filtrate under reduced pressure to give mixtures 6 and 6' from the product1The H NMR results judged the ratio of compound 6 to isomer 6' to be 1: 0.28.
comparative example No. four
Compound 5 of the present invention is subjected to Raney Ni/H2Reducing and synthesizing the cholesterol. Shown in the following equation eight:
at the same time, the present inventors tried H2As a reducing agent, 10% Pd/C as a catalyst to perform selective hydrogenation reduction reaction on the double bonds of the side chains. The reaction results are shown in the formula nine, and compounds 8 and 8' are obtained (the conditions of TLC detection of the reaction solution are shown in FIG. 6), but the expected target product cholesterol is not obtained, which indicates that 10% Pd/C can not replace Raney Ni as a catalyst to perform selective hydrogenation reduction on the side chain.
The experimental method comprises the following steps: a100 mL single neck flask was charged with compound of formula (5) (1.50g, 3.56mmol), 10% Pd/C (150mg), isopropanol (30mL), H2(1atm), reaction at 30 ℃ for 7h, TLC monitoring the completion of the reaction, filtering to remove Pd/C, and concentrating the filtrate under reduced pressure to obtain reaction formula 8 and isomer 8'. From the product1The H NMR results judged the ratio of compound 8 to isomer 8' to be 1: 0.25.
the protection of the present invention is not limited to the above embodiments. Variations and advantages that may occur to those skilled in the art may be incorporated into the invention without departing from the spirit and scope of the inventive concept, which is set forth in the following claims.
Claims (12)
1. A method for synthesizing cholesterol by taking BA as a raw material is characterized in that the method takes BA as a raw material and synthesizes the cholesterol through the steps of oxidation, Wittig reaction, acetylation, reduction and selective hydrogenation reduction, and specifically comprises the following steps:
step (a), in a first solvent, performing oxidation reaction on BA shown in a formula (1) to obtain a compound shown in a formula (2);
in a second solvent, carrying out a Wittig reaction on the compound shown in the formula (2) to obtain a compound shown in the formula (3);
in a third solvent, performing acetylation reaction on the compound in the formula (3) to obtain a compound in a formula (4);
in a fourth solvent, carrying out reduction reaction on the compound shown in the formula (4) to obtain compounds shown in the formula (5) and the formula (6);
in a fifth solvent, carrying out selective hydrogenation reduction reaction on the compound shown in the formula (5) to obtain the cholesterol;
wherein the reaction process of the method is shown as a scheme (A):
2. the method according to claim 1, wherein in step (a), the oxidation reaction is specifically: in the first solvent, BA shown in the formula (1) is subjected to oxidation reaction with TEMPO, sodium bicarbonate, tetrabutylammonium bromide and an oxidant to obtain a compound shown in the formula (2).
3. The method according to claim 2, wherein the molar ratio of BA, TEMPO, sodium bicarbonate, tetrabutylammonium bromide and the oxidizing agent represented by formula (1) is 1: (0.01-1): (1.35-20): (0.1-1): (1.15-5); and/or the oxidant is selected from one or more of N-chlorosuccinimide NCS, N-bromosuccinimide NBS and 2-iodosylbenzoic acid IBX; and/or, the first solvent is selected from one or more of dichloromethane, tetrahydrofuran, toluene, dimethyl sulfoxide and water; and/or the temperature of the oxidation reaction is 0-30 ℃; and/or the time of the oxidation reaction is 3-8 h.
4. The method as claimed in claim 1, wherein in step (b), the Wittig reaction is specifically: in the second solvent, the compound of the formula (2), the 3, 3-dimethylallyl halide, triphenylphosphine and potassium tert-butoxide are subjected to a Wittig reaction to obtain the compound of the formula (3).
5. The process of claim 4, wherein the molar ratio of the compound of formula (2), 3-dimethylallyl halide, triphenylphosphine, potassium tert-butoxide is 1: (1.5-4): (1.5-4): (1-4); and/or the second solvent is one or more of toluene, benzene, tetrahydrofuran and n-hexane; and/or the temperature of the Wittig reaction is 60-140 ℃; and/or the Wittig reaction time is 4-9 h.
6. The process according to claim 1, wherein in step (c), the acetylation reaction is in particular: and (3) performing acetylation reaction on the compound of the formula (3), acetyl chloride, acetic anhydride and pyridine in the third solvent to obtain the compound of the formula (4).
7. The process of claim 6, wherein the compound of formula (3), acetyl chloride, acetic anhydride, pyridine are present in a molar ratio of 1: (25-62.5): (25-62.5): (4-6); and/or the third solvent is one or more of acetic anhydride, acetyl chloride, ethyl acetate and dichloromethane; and/or the temperature of the acetylation reaction is 40-110 ℃; and/or the acetylation reaction time is 3-5 h.
8. The method according to claim 1, wherein in step (d), the reduction reaction is specifically: and (3) carrying out reduction reaction on the compound of the formula (4) and a reducing agent in the fourth solvent to obtain compounds of the formula (5) and the formula (6).
9. The method according to claim 8, wherein the molar ratio of the compound of formula (4) to the reducing agent is 1 (15-25); and/or the fourth solvent is one or more of tetrahydrofuran, ethanol, water, dichloromethane, 2-methyltetrahydrofuran, isopropanol, acetic acid and methyl tert-butyl ether; and/or, the reducing agent is selected from NaBH4、KBH4One or two of them; and/or the temperature of the reduction reaction is 0-30 ℃; and/or the time of the reduction reaction is 6-9 h.
10. The method according to claim 1, wherein in step (e), the selective hydrogenation reduction reaction is specifically: and (3) carrying out selective hydrogenation reduction reaction on the compound shown in the formula (5) and a reducing agent in the fifth solvent under the action of a catalyst to obtain cholesterol.
11. The method of claim 10, wherein the reducing agent is selected from H2(ii) a And/or the catalyst is RaneyNi; and/or the mass ratio of the compound of the formula (5) to the catalyst is 1: (0.05-5); and/or the fifth solvent is selected from one or more of isopropanol, dichloromethane, methanol, 2-methyltetrahydrofuran, tetrahydrofuran, ethanol, water, methyl tert-butyl ether, ethyl acetate and toluene; and/or the temperature of the hydrogenation reduction reaction is 0-60 ℃; and/or, the reducing agent H of the hydrogenation reduction reaction2The pressure is 1-20 atm; and/or the time of the hydrogenation reduction reaction is 6-10 h.
12. A compound, characterized in that the structure of the compound is represented by formula (4):
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| CN114395009A (en) * | 2022-01-27 | 2022-04-26 | 华东师范大学 | Synthesis method of high-purity cholesterol |
| CN114874277A (en) * | 2022-03-08 | 2022-08-09 | 江苏佳尔科药业集团股份有限公司 | Synthesis method of cholesterol |
| WO2022213805A1 (en) * | 2021-04-09 | 2022-10-13 | 华东师范大学 | Method for synthesizing cholesterol by using ba as raw material |
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| CN114524856B (en) * | 2022-01-27 | 2024-03-15 | 华东师范大学 | Synthesis method of high-purity plant-derived cholesterol |
| WO2023152768A1 (en) * | 2022-02-11 | 2023-08-17 | Fermenta Biotech Limited | Synthesis of cholesterol form bisnoralcohol |
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| CN111072744A (en) * | 2019-12-03 | 2020-04-28 | 华东师范大学 | Method for synthesizing ursodeoxycholic acid by taking BA as raw material |
| CN111560045A (en) * | 2020-06-23 | 2020-08-21 | 江苏佳尔科药业集团股份有限公司 | Method for synthesizing lithocholic acid by taking BA as raw material |
| CN112608361A (en) * | 2019-12-19 | 2021-04-06 | 湖南科瑞生物制药股份有限公司 | Process for preparing cholesterol, derivatives and analogues thereof |
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| US5714481A (en) * | 1983-08-02 | 1998-02-03 | Research Corporation Technologies, Inc. | Derivatives of 5-androsten-17 ones and 5-androstan-17-ones |
| JPS61129196A (en) * | 1984-11-27 | 1986-06-17 | Chugai Pharmaceut Co Ltd | Production of 3beta-hydroxy-5-cholic acid |
| JPS61227592A (en) * | 1985-04-01 | 1986-10-09 | Chugai Pharmaceut Co Ltd | 27-nor-25-oxocholesterol |
| CN104961788A (en) * | 2015-06-25 | 2015-10-07 | 湖南科瑞生物科技股份有限公司 | Synthetic method of cholesterol |
| CN106632565A (en) * | 2016-11-07 | 2017-05-10 | 湖南科瑞生物制药股份有限公司 | Novel method used for synthesizing cholesterol |
| CN113248557A (en) * | 2021-04-09 | 2021-08-13 | 华东师范大学 | Method for synthesizing cholesterol by taking BA as raw material |
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| CN111072744A (en) * | 2019-12-03 | 2020-04-28 | 华东师范大学 | Method for synthesizing ursodeoxycholic acid by taking BA as raw material |
| CN112608361A (en) * | 2019-12-19 | 2021-04-06 | 湖南科瑞生物制药股份有限公司 | Process for preparing cholesterol, derivatives and analogues thereof |
| CN111560045A (en) * | 2020-06-23 | 2020-08-21 | 江苏佳尔科药业集团股份有限公司 | Method for synthesizing lithocholic acid by taking BA as raw material |
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| WO2022213805A1 (en) * | 2021-04-09 | 2022-10-13 | 华东师范大学 | Method for synthesizing cholesterol by using ba as raw material |
| CN114395009A (en) * | 2022-01-27 | 2022-04-26 | 华东师范大学 | Synthesis method of high-purity cholesterol |
| WO2023142890A1 (en) * | 2022-01-27 | 2023-08-03 | 华东师范大学 | Synthesis method for high-purity cholesterol |
| CN114395009B (en) * | 2022-01-27 | 2024-02-27 | 华东师范大学 | High-purity cholesterol synthesis method |
| CN114874277A (en) * | 2022-03-08 | 2022-08-09 | 江苏佳尔科药业集团股份有限公司 | Synthesis method of cholesterol |
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