CA2705902A1 - A method for producing betulinic acid - Google Patents
A method for producing betulinic acid Download PDFInfo
- Publication number
- CA2705902A1 CA2705902A1 CA2705902A CA2705902A CA2705902A1 CA 2705902 A1 CA2705902 A1 CA 2705902A1 CA 2705902 A CA2705902 A CA 2705902A CA 2705902 A CA2705902 A CA 2705902A CA 2705902 A1 CA2705902 A1 CA 2705902A1
- Authority
- CA
- Canada
- Prior art keywords
- betulinic
- acid
- betulin
- betulinic acid
- aldehyde
- 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.)
- Granted
Links
- QGJZLNKBHJESQX-FZFNOLFKSA-N betulinic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CC[C@@H](C(=C)C)[C@@H]5[C@H]4CC[C@@H]3[C@]21C QGJZLNKBHJESQX-FZFNOLFKSA-N 0.000 title claims abstract description 85
- QGJZLNKBHJESQX-UHFFFAOYSA-N 3-Epi-Betulin-Saeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C(=C)C)C5C4CCC3C21C QGJZLNKBHJESQX-UHFFFAOYSA-N 0.000 title claims abstract description 76
- CLOUCVRNYSHRCF-UHFFFAOYSA-N 3beta-Hydroxy-20(29)-Lupen-3,27-oic acid Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C(O)=O)CCC5(C)CCC(C(=C)C)C5C4CCC3C21C CLOUCVRNYSHRCF-UHFFFAOYSA-N 0.000 title claims abstract description 76
- DIZWSDNSTNAYHK-XGWVBXMLSA-N Betulinic acid Natural products CC(=C)[C@@H]1C[C@H]([C@H]2CC[C@]3(C)[C@H](CC[C@@H]4[C@@]5(C)CC[C@H](O)C(C)(C)[C@@H]5CC[C@@]34C)[C@@H]12)C(=O)O DIZWSDNSTNAYHK-XGWVBXMLSA-N 0.000 title claims abstract description 76
- PZXJOHSZQAEJFE-UHFFFAOYSA-N dihydrobetulinic acid Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C(C)C)C5C4CCC3C21C PZXJOHSZQAEJFE-UHFFFAOYSA-N 0.000 title claims abstract description 76
- MQYXUWHLBZFQQO-UHFFFAOYSA-N nepehinol Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C)CCC(C(=C)C)C5C4CCC3C21C MQYXUWHLBZFQQO-UHFFFAOYSA-N 0.000 title claims abstract description 76
- 238000004519 manufacturing process Methods 0.000 title abstract description 14
- FELCJAPFJOPHSD-ROUWMTJPSA-N Lup-20(29)-en-28-al, 3beta-hydroxy- Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C=O)CC[C@@H](C(=C)C)[C@@H]5[C@H]4CC[C@@H]3[C@]21C FELCJAPFJOPHSD-ROUWMTJPSA-N 0.000 claims abstract description 50
- FVWJYYTZTCVBKE-ROUWMTJPSA-N betulin Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(CO)CC[C@@H](C(=C)C)[C@@H]5[C@H]4CC[C@@H]3[C@]21C FVWJYYTZTCVBKE-ROUWMTJPSA-N 0.000 claims abstract description 44
- MVIRREHRVZLANQ-UHFFFAOYSA-N betulin Natural products CC(=O)OC1CCC2(C)C(CCC3(C)C2CC=C4C5C(CCC5(CO)CCC34C)C(=C)C)C1(C)C MVIRREHRVZLANQ-UHFFFAOYSA-N 0.000 claims abstract description 44
- JYDNKGUBLIKNAM-UHFFFAOYSA-N Oxyallobutulin Natural products C1CC(=O)C(C)(C)C2CCC3(C)C4(C)CCC5(CO)CCC(C(=C)C)C5C4CCC3C21C JYDNKGUBLIKNAM-UHFFFAOYSA-N 0.000 claims abstract description 43
- MHAVMNJPXLZEIG-UHFFFAOYSA-N betulinic aldehyde Natural products C1CC(=O)C(C)(C)C2CCC3(C)C4(C)CCC5(C=O)CCC(C(=C)C)C5C4CCC3C21C MHAVMNJPXLZEIG-UHFFFAOYSA-N 0.000 claims abstract description 43
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 23
- 238000000034 method Methods 0.000 claims abstract description 19
- 238000002360 preparation method Methods 0.000 claims abstract description 12
- 230000003647 oxidation Effects 0.000 claims abstract description 11
- 238000001953 recrystallisation Methods 0.000 claims abstract description 5
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 57
- 239000000203 mixture Substances 0.000 claims description 45
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 36
- 239000000243 solution Substances 0.000 claims description 35
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 32
- NDVLTYZPCACLMA-UHFFFAOYSA-N silver oxide Chemical compound [O-2].[Ag+].[Ag+] NDVLTYZPCACLMA-UHFFFAOYSA-N 0.000 claims description 26
- 238000003756 stirring Methods 0.000 claims description 25
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 20
- 239000003960 organic solvent Substances 0.000 claims description 19
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 18
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 14
- 229910001923 silver oxide Inorganic materials 0.000 claims description 13
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 12
- 239000007800 oxidant agent Substances 0.000 claims description 12
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 11
- JRNVZBWKYDBUCA-UHFFFAOYSA-N N-chlorosuccinimide Chemical compound ClN1C(=O)CCC1=O JRNVZBWKYDBUCA-UHFFFAOYSA-N 0.000 claims description 11
- 239000012670 alkaline solution Substances 0.000 claims description 11
- 239000012044 organic layer Substances 0.000 claims description 11
- 239000010410 layer Substances 0.000 claims description 9
- YRIZYWQGELRKNT-UHFFFAOYSA-N 1,3,5-trichloro-1,3,5-triazinane-2,4,6-trione Chemical group ClN1C(=O)N(Cl)C(=O)N(Cl)C1=O YRIZYWQGELRKNT-UHFFFAOYSA-N 0.000 claims description 8
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 8
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 8
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 8
- 230000001590 oxidative effect Effects 0.000 claims description 7
- 230000020477 pH reduction Effects 0.000 claims description 5
- 239000000376 reactant Substances 0.000 claims description 5
- 230000002194 synthesizing effect Effects 0.000 claims description 5
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 4
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 4
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 4
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 4
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 4
- 239000007858 starting material Substances 0.000 abstract 1
- 238000010189 synthetic method Methods 0.000 abstract 1
- 239000005457 ice water Substances 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 7
- 239000000706 filtrate Substances 0.000 description 7
- 239000012141 concentrate Substances 0.000 description 5
- 235000019439 ethyl acetate Nutrition 0.000 description 5
- 238000001914 filtration Methods 0.000 description 4
- 230000000975 bioactive effect Effects 0.000 description 3
- 239000013078 crystal Substances 0.000 description 3
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000012264 purified product Substances 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 239000001488 sodium phosphate Substances 0.000 description 2
- 229910000162 sodium phosphate Inorganic materials 0.000 description 2
- 238000001308 synthesis method Methods 0.000 description 2
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 2
- 208000030507 AIDS Diseases 0.000 description 1
- 244000274847 Betula papyrifera Species 0.000 description 1
- 235000009113 Betula papyrifera Nutrition 0.000 description 1
- 235000009109 Betula pendula Nutrition 0.000 description 1
- 235000010928 Betula populifolia Nutrition 0.000 description 1
- 235000002992 Betula pubescens Nutrition 0.000 description 1
- SLJTWDNVZKIDAU-CKURCAGRSA-N Betulonic acid Natural products CC(=C)[C@@H]1CC[C@@]2(CC[C@]3(C)[C@@H](CC[C@@H]4[C@@]5(C)CCC(=O)C(C)(C)[C@@H]5CC[C@@]34C)[C@@H]12)C(=O)O SLJTWDNVZKIDAU-CKURCAGRSA-N 0.000 description 1
- SLJTWDNVZKIDAU-SVAFSPIFSA-N Betulonic acid Chemical compound C1CC(=O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CC[C@@H](C(=C)C)[C@@H]5[C@H]4CC[C@@H]3[C@]21C SLJTWDNVZKIDAU-SVAFSPIFSA-N 0.000 description 1
- 101150041968 CDC13 gene Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 238000006809 Jones oxidation reaction Methods 0.000 description 1
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 1
- ZNUAKACHFYTNFX-UHFFFAOYSA-N Wallichenol Natural products CC12CCC(O)C(C)(C)C1CCC1(C)C2CCC2C3C(C(=C)CO)CCC3(C)CCC21C ZNUAKACHFYTNFX-UHFFFAOYSA-N 0.000 description 1
- 238000006136 alcoholysis reaction Methods 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000036436 anti-hiv Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000078 anti-malarial effect Effects 0.000 description 1
- 239000003430 antimalarial agent Substances 0.000 description 1
- -1 betulinol Chemical compound 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 229930000223 plant secondary metabolite Natural products 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011946 reduction process Methods 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
- 238000007039 two-step reaction Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
A method for producing betulinic acid using betulin as the starting material comprises the following steps: (1) Preparation of crude betulinic aldehyde through oxidation of betulin; (2) Preparation of crude betulinic acid through oxidation of crude betulinic aldehyde; and (3) Preparation of betulinic acid through recrystallization of crude betulinic acid. The present invention changes the disadvantages existing in the previous synthetic methods of betulinic acid that betulinic acid isomer occurs in the product or the routing is complex.
The method of present invention has a low producing cost.
The method of present invention has a low producing cost.
Description
A Method for Producing Betulinic Acid FIELD OF THE INVENTION
The present invention relates to a semi-synthesis method for producing natural compound, and in particular, to a semi-synthesis method for producing betulinic acid from betulin.
BACKGROUND OF THE INVENTION
Betulinic acid (I), also known as R-betulinic acid, is a plant secondary metabolite of pentacyclic triterpene, which is a white needle crystal with Molecular formula of C30H4803, molecular weight of 456.71; and structural formula:
Jr Ji jeV
H H
21Ot-I H OH
HO HO
I II
Betulinic acid has antimalarial, anti-inflammatory and anti-HIV activity, and displays cytotoxic activities to melanoma cells, which makes it as an anticancer and anti-AIDS compound with broad application and developmental prospect. The content of betulinic acid in plants is limited, while the content of betulin (i.e. betulinol, II), a betulinic acid analogue, in white birch bark is very high, reaching 1035%. Betulinic acid may be produced through semi-synthesis from betulin.
The United States Patent and Trademark Office disclosed a series of invention patents entitled "Methods for manufacturing betulinic acid" on May 15, 2001, August 7, 2001, June 18, 2002 and March 15, 2005, respectively, which provide a five-step method for synthesizing betulinic acid from betulin:
firstly, acylating betulin into betulin 3,28- dicarboxylic ester; secondly, converting betulin 3,28- dicarboxylic ester into betulin-3- acetic ester through alcoholysis;
thirdly, oxidizing betulin-3-acetic ester into betulinic aldehyde-3-acetic ester, and then betulinic acid-3-acetic ester; finally, deprotecting to generate betulinic acid. This method has long reaction route, complex reaction conditions and high producing cost.
On September 8, 1998, The United States Patent and Trademark Office disclosed a patent of invention entitled "Methods of Manufacturing betulinic acid", which provides a process for synthesizing betulinic acid by a two-step reaction: firstly, forming betulonic acid intermediate through Jones oxidation, and then generating betulinic acid through a selective reduction reaction.
This method is simple, but what is generated after the reduction process is a mixture of betulinic acid and a-betulinic acid, a non-bioactive betulinic acid isomer. Through recrystallization, this product still contains a-betulinic acid.
SUMMARY OF THE INVENTION
The present invention is designed to provide a method for producing betulinic acid, with simple routing and easily purified product.
In order to achieve the object of the present invention, in an aspect, the present invention provides a method for generating betulinic acid from betulin through twice selective oxidations.
Wherein, the selective oxidation reaction is to oxidize C-28 hydroxyl group of betulin.
In particular, the selective oxidation reaction comprises: step (1) Preparation of betulinic aldehyde through selective oxidation of betulin; and step (2) Preparation of betulinic acid through oxidation reaction of betulinic aldehyde.
Wherein, the oxidizer for the selective oxidation reaction in step (1) is trichloroiminocyanuric acid or N-chlorosuccinimide; and the oxidizer for the selective oxidation reaction in step (2) is silver oxide.
In particular, the selective oxidation reaction also comprises the purification of betulinic acid by recrystallizing the crude betulinic acid prepared through oxidation reaction.
In a further aspect, the present invention provides a method for twice selective oxidization of betulin, wherein, the oxidization reaction comprises:
oxidizing betulin into betulinic aldehyde first; then oxidizing betulinic aldehyde into crude betulinic acid; and finally, recrystallizing the crude betulinic acid to obtain betulinic acid.
In still a further aspect, the present invention provides a method for producing betulinic acid, comprising the following steps:
(1) Preparation of betulinic aldehyde through oxidation of betulin:
Firstly, dissolve betulin into an organic solvent, and add 2,2,6,6-tetramethyl-1- piperidone, alkaline solution and oxidizer into the mixture one by one; then make the mixture react for 0.5-12 hours at 2575 C
under stirring and produce betulinic aldehyde;
(2) Preparation of betulinic acid through oxidation of betulinic aldehyde:
Firstly, dissolve betulinic aldehyde into an organic solvent, and add sodium hydroxide solution and silver oxide into the mixture one by one; then make the mixture react for 3-72 hours at 2585 C under stirring, and prepare crude betulinic acid through acidification.
Wherein, the molar ratio of betulin and 2,2,6,6-tetramethyl-1-piperidone mentioned in step (1) is 1:0.01-0.05; and the molar ratio of betulin and the oxidizer is 1: 0.8-1.5; the organic solvent dosage is 100-200ml per gram of betulin; and the alkaline solution dosage is 100-200ml per gram of betulin.
In particular, the organic solvent mentioned in step (1) is selected from one of methylene dichloride, chloroform or ethyl acetate; the alkaline solution is selected from one of sodium hydroxide solution, sodium carbonate solution or sodium bicarbonate solution; and the oxidizer is trichloroiminocyanuric acid or N-chlorosuccinimide.
Wherein, the pH value of the alkaline solution is 8.3-12.5;
In particular, the step (1) also comprises: standing to layer the reactant, separating the organic layer and evaporating the organic solvent by vacuum so as to prepare the betulinic aldehyde.
Wherein, mentioned in step (2) the molar ratio of betulinic aldehyde and silver oxide is 1: 0.8-1.5. The organic solvent is selected from one of tetrahydrofuran, dioxane, ethanol or acetone; and the mass concentration of sodium hydroxide solution is 10-40%, 30% being preferable.
In particular, the organic solvent dosage is 5-20m1 per gram of betulinic aldehyde; sodium hydroxide solution dosage is 0.5 - 1.5 ml per gram of betulinic aldehyde, and silver oxide is prepared freshly.
In particular, the step (2) also comprises: standing to precipitate the reaction solution of crude betulinic acid upon reaction first, filtering the reactant and then recrystallizing the sediment obtained by filtering so as to provide the betulinic acid.
Preferably, adding the acidified solution into ice water, standing to precipitate the mixture, and then conducting the above-mentioned filtration.
In particular, step (2) also comprises: extracting the reaction solution of crude betulinic acid upon reaction first, and then concentrating and recrystallizing the sediment so as to obtain betulinic acid.
Preferably, adding the acidified solution into ice water, and then conducting the above-mentioned extraction.
Wherein, a concentrated hydrochloric acid is adopted for the acidification, and the pH value of the acidified solution is 2.0-3.0, and 2.5 being preferable;
the mass concentration of the concentrated hydrochloric acid is 20-37%; the methylene dichloride are adopted for the extraction of the acidified solution;
one of ethanol, methanol or acetic acid is adopted for the recrystallization.
In particular, betulinic acid is prepared by concentrating organic layer upon extraction.
The present invention relates to a semi-synthesis method for producing natural compound, and in particular, to a semi-synthesis method for producing betulinic acid from betulin.
BACKGROUND OF THE INVENTION
Betulinic acid (I), also known as R-betulinic acid, is a plant secondary metabolite of pentacyclic triterpene, which is a white needle crystal with Molecular formula of C30H4803, molecular weight of 456.71; and structural formula:
Jr Ji jeV
H H
21Ot-I H OH
HO HO
I II
Betulinic acid has antimalarial, anti-inflammatory and anti-HIV activity, and displays cytotoxic activities to melanoma cells, which makes it as an anticancer and anti-AIDS compound with broad application and developmental prospect. The content of betulinic acid in plants is limited, while the content of betulin (i.e. betulinol, II), a betulinic acid analogue, in white birch bark is very high, reaching 1035%. Betulinic acid may be produced through semi-synthesis from betulin.
The United States Patent and Trademark Office disclosed a series of invention patents entitled "Methods for manufacturing betulinic acid" on May 15, 2001, August 7, 2001, June 18, 2002 and March 15, 2005, respectively, which provide a five-step method for synthesizing betulinic acid from betulin:
firstly, acylating betulin into betulin 3,28- dicarboxylic ester; secondly, converting betulin 3,28- dicarboxylic ester into betulin-3- acetic ester through alcoholysis;
thirdly, oxidizing betulin-3-acetic ester into betulinic aldehyde-3-acetic ester, and then betulinic acid-3-acetic ester; finally, deprotecting to generate betulinic acid. This method has long reaction route, complex reaction conditions and high producing cost.
On September 8, 1998, The United States Patent and Trademark Office disclosed a patent of invention entitled "Methods of Manufacturing betulinic acid", which provides a process for synthesizing betulinic acid by a two-step reaction: firstly, forming betulonic acid intermediate through Jones oxidation, and then generating betulinic acid through a selective reduction reaction.
This method is simple, but what is generated after the reduction process is a mixture of betulinic acid and a-betulinic acid, a non-bioactive betulinic acid isomer. Through recrystallization, this product still contains a-betulinic acid.
SUMMARY OF THE INVENTION
The present invention is designed to provide a method for producing betulinic acid, with simple routing and easily purified product.
In order to achieve the object of the present invention, in an aspect, the present invention provides a method for generating betulinic acid from betulin through twice selective oxidations.
Wherein, the selective oxidation reaction is to oxidize C-28 hydroxyl group of betulin.
In particular, the selective oxidation reaction comprises: step (1) Preparation of betulinic aldehyde through selective oxidation of betulin; and step (2) Preparation of betulinic acid through oxidation reaction of betulinic aldehyde.
Wherein, the oxidizer for the selective oxidation reaction in step (1) is trichloroiminocyanuric acid or N-chlorosuccinimide; and the oxidizer for the selective oxidation reaction in step (2) is silver oxide.
In particular, the selective oxidation reaction also comprises the purification of betulinic acid by recrystallizing the crude betulinic acid prepared through oxidation reaction.
In a further aspect, the present invention provides a method for twice selective oxidization of betulin, wherein, the oxidization reaction comprises:
oxidizing betulin into betulinic aldehyde first; then oxidizing betulinic aldehyde into crude betulinic acid; and finally, recrystallizing the crude betulinic acid to obtain betulinic acid.
In still a further aspect, the present invention provides a method for producing betulinic acid, comprising the following steps:
(1) Preparation of betulinic aldehyde through oxidation of betulin:
Firstly, dissolve betulin into an organic solvent, and add 2,2,6,6-tetramethyl-1- piperidone, alkaline solution and oxidizer into the mixture one by one; then make the mixture react for 0.5-12 hours at 2575 C
under stirring and produce betulinic aldehyde;
(2) Preparation of betulinic acid through oxidation of betulinic aldehyde:
Firstly, dissolve betulinic aldehyde into an organic solvent, and add sodium hydroxide solution and silver oxide into the mixture one by one; then make the mixture react for 3-72 hours at 2585 C under stirring, and prepare crude betulinic acid through acidification.
Wherein, the molar ratio of betulin and 2,2,6,6-tetramethyl-1-piperidone mentioned in step (1) is 1:0.01-0.05; and the molar ratio of betulin and the oxidizer is 1: 0.8-1.5; the organic solvent dosage is 100-200ml per gram of betulin; and the alkaline solution dosage is 100-200ml per gram of betulin.
In particular, the organic solvent mentioned in step (1) is selected from one of methylene dichloride, chloroform or ethyl acetate; the alkaline solution is selected from one of sodium hydroxide solution, sodium carbonate solution or sodium bicarbonate solution; and the oxidizer is trichloroiminocyanuric acid or N-chlorosuccinimide.
Wherein, the pH value of the alkaline solution is 8.3-12.5;
In particular, the step (1) also comprises: standing to layer the reactant, separating the organic layer and evaporating the organic solvent by vacuum so as to prepare the betulinic aldehyde.
Wherein, mentioned in step (2) the molar ratio of betulinic aldehyde and silver oxide is 1: 0.8-1.5. The organic solvent is selected from one of tetrahydrofuran, dioxane, ethanol or acetone; and the mass concentration of sodium hydroxide solution is 10-40%, 30% being preferable.
In particular, the organic solvent dosage is 5-20m1 per gram of betulinic aldehyde; sodium hydroxide solution dosage is 0.5 - 1.5 ml per gram of betulinic aldehyde, and silver oxide is prepared freshly.
In particular, the step (2) also comprises: standing to precipitate the reaction solution of crude betulinic acid upon reaction first, filtering the reactant and then recrystallizing the sediment obtained by filtering so as to provide the betulinic acid.
Preferably, adding the acidified solution into ice water, standing to precipitate the mixture, and then conducting the above-mentioned filtration.
In particular, step (2) also comprises: extracting the reaction solution of crude betulinic acid upon reaction first, and then concentrating and recrystallizing the sediment so as to obtain betulinic acid.
Preferably, adding the acidified solution into ice water, and then conducting the above-mentioned extraction.
Wherein, a concentrated hydrochloric acid is adopted for the acidification, and the pH value of the acidified solution is 2.0-3.0, and 2.5 being preferable;
the mass concentration of the concentrated hydrochloric acid is 20-37%; the methylene dichloride are adopted for the extraction of the acidified solution;
one of ethanol, methanol or acetic acid is adopted for the recrystallization.
In particular, betulinic acid is prepared by concentrating organic layer upon extraction.
Advantages of the present invention:
1. The present invention is to generate crude betulinic aldehyde by oxidizing betulin and then oxidize crude betulinic aldehyde, so that, in the oxidation product, only bioactive betulinic acid is generated, without non-bioactive a-betulinic acid.
2. The present invention adopts a step-by-step oxidization and highly-selective oxidizer which has no effect on C-3 hydroxyl group, thus functional group protection is not necessary.
3. The present invention has the superiorities of short routing, simple processing method, easily purified product and low production cost, thus industrialized production and popularization is easily achieved.
DESCRIPTION OF DRAWINGS
Fig.1 Schematic diagram for synthesizing betulinic acid through oxidization of betulin DETAILED DESCRIPTION OF THE INVENTION
Further details of the invention are described by combining the Examples of the present invention, as follows:
A method for producing betulinic acid, with the betulin as the raw material, comprising the following steps:
(1) Preparation of crude betulinic aldehyde through oxidation of betulin:
dissolve betulin into an organic solvent, and add 2,2,6,6-tetramethyl-1-piperidone, inorganic alkaline solution with a pH value of 8.012.5 and oxidizer into the mixture; stir to make the mixture react at 2575 C for 0.5-12 hours; upon completion of reaction, keep the mixture standing to layer, separate the organic layer and make the organic phase concentrated so as to provide the grey white crude betulinic aldehyde; the molar ratio of betulin and 2,2,6,6-tetramethyl-1-piperidone is 1:0.01-0.05, and the molar ratio of betulin and the oxidizer is 1: 0.8-1.5; the organic solvent dosage is 100-200ml per gram of betulin; and the inorganic alkaline solution dosage is 100-200ml per gram of betulin;
(2) Preparation of crude betulinic acid through oxidation of crude betulinic aldehyde: dissolve crude betulinic aldehyde generated in step (1) into an organic solvent, and add sodium hydroxide solution and the freshly prepared silver oxide into the mixture; after stirring to make the mixture react at for 3-72 hours, make the reactant cool to room temperature, and filter the reactant; The filtrate is generated betulinic acid through acidification, filtering, washing and drying; the organic solvent dosage is 5-20m1 per gram of crude betulinic aldehyde;
(3) Preparation of betulinic acid through recrystallization of crude betulinic acid: recrystallizing the crude betulinic acid obtained in step (2) by using ethanol, methanol or acetic acid as the solvent to obtain betulinic acid.
The method for producing betulinic acid, wherein, the organic solvent mentioned in step (1) is methylene dichloride, chloroform or ethyl acetate.
The method for producing betulinic acid, wherein, the inorganic alkaline solution mentioned in step (1) is sodium hydroxide solution, sodium phosphate, sodium carbonate solution or sodium bicarbonate solution.
The method for producing betulinic acid, wherein, the oxidizer mentioned in step (1) is trichloroiminocyanuric acid or N-chlorosuccinimide.
The method for producing betulinic acid, wherein, the organic solvent mentioned in step (2) is tetrahydrofuran, dioxane, ethanol or acetone.
Embodiment 1 Dissolve 44.3g (0.1 moL) betulin (I) into 5000m1 methylene dichloride, and add 0.156g (0.001 moL) 2,2,6,6-tetramethyl-1-piperidone, 5000m1 sodium bicarbonate solution with a pH value of 8.3, and 20g (0.15moL) N-chlorosuccinimide one by one while stirring; after reacting at 25 C for 10 hours under stirring, stand to layer the mixture, separate the organic layer, and evaporate methylene dichloride under vacuum so as to yield 45.5g crude betulinic aldehyde (III); dissolve the obtained crude betulinic aldehyde into 910m1 ethanol, add 50ml sodium hydroxide solution with a mass concentration of 30%, and then add 24g freshly prepared silver oxide into the mixture; after react at 70 C under stirring for 5 hours, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 36%) into the filtrate until the pH value is 2.5, and then pour the mixture into 5000m1 ice water, extract with methylene dichloride and concentrate so as to provide crude betulinic acid; and recrystallize crude betulinic acid with methanol so as to yield 23.6g betulinic acid (II) with a purity of 92.9% and yield of 48%.
Betulinic aldehyde is white crystal (methanol); melting point: 190-192 C
(192-193 C); El-MS m/z: 440([M+]), 207, 189, 135; 'H-NMR(300MHz , CDCI3);60.76 (s, 3H, 23-Me) , 0.83(s, 3H, 24-Me), 0.92 (s, 3H, 25-Me), 0.97(s, 3H, 26-Me), 0.98(s,3H,27-Me),1.70(s, 3H, 30-Me),3.19 (q, 3-H), 4.76 (s, 29a-H), 4.63(s, 29b-H), 9.68(s, 28-H);13C-NMR(300MHz, CDCI3038.9(C-1), 28.0(C-2), 79.0(C-3), 40.8(C-4), 59.4(C-5), 18.3(C-6), 34.3(C-7), 40.8(C-8), 50.5(C-9), 37.2(C-10), 20.8(C-11), 25.6(C-12), 38.7(C-13),42.6 (C-14), 29.9 (C-15), 34.3 (C-16), 55.3 (C-17), 47.6 (C-18), 48.1 (C-19), 149.8(C-20), 29.9(C-21), 38.7(C-22), 27.4 (C-23), 15.9(C-24), 16.2(C-25), 16.2(C-26), 14.3(C-27), 206.7(C-28), 110.2 (C-29), 19.0 (C-30).
The betulinic acid is white crystal (methanol); melting point:
284-287 C(285-287 C); El-MS m/z: 456([M+]), 248, 220, 207, 189, 175;
1H-NMR(300MHz,CDCI3) :O0.76(s, 3H, 23-Me), 0.83(s, 3H, 24-Me), 0.94(s, 3H, 25-Me), 0.97(s, 3H, 26-Me), 0.98(s, 3H, 27-Me), 1.69(s, 3H, 30-Me), 3.17(q, 3-H), 4.74(s, 29a-H), 4.62(s, 29b-H); 13C-NMR(300MHz, CDC13): 638.4(C-1), 28.0(C-2), 79.0(C-3), 40.7(C-4), 55.4(C-5), 18.3(C-6), 34.3(C-7), 38.4(C-8), 50.5(C-9), 37.0(C-10), 19.3(C-11), 27.4(C-12), 38.7(C-13), 42.5(C-14), 29.7(C-15), 34.3(C-16), 56.3(C-17), 49.3(C-18), 46.9(C-19), 150.4(C-20), 29.7(C-21), 37.2(C-22), 28.0(C-23), 15.4(C-24), 16.0(C-25), 16.1(C-26), 14.7(C-27), 180.7(C-28),109.7(C-29), 19.4(C-30).
Embodiment 2 Dissolve 44.3g (0.1 moL) betulin (I) into 4430ml chloroform, and add 0.78g (0.005 moL) 2,2,6,6-tetramethyl-1-piperidone, 4430m1 sodium phosphate solution with a pH value of 11.5, and 26g (0.11 moL) trichloroiminocyanuric acid one by one while stirring; after reacting at 50 C for 0.5 hours under stirring, stand to layer the mixture and separate the organic layer, evaporate chloroform under vacuum so as to yield 44.Og crude betulinic aldehyde (III).
Dissolve crude betulinic aldehyde into 220m1 tetrahydrofuran, add 50m1 sodium hydroxide solution with a mass concentration of 30%, and then add 24g freshly prepared silver oxide into the mixture; after reacting at 40 C
under stirring for 30 hours, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 30%) into the filtrate until the pH value is 2.5, and then pour the mixture into 5000m1 ice water, extract with methylene dichloride and concentrate so as to provide crude betulinic acid; and recrystallize crude betulinic acid with ethanol so as to yield 22.6g betulinic acid (II) with a purity of 93.1% and yield of 46%.
Embodiment 3 Dissolve 44.3g (0.1 moL) betulin (I) into 5000m1 methylene dichloride, and add 0.25g (0.0016 moL) 2,2,6,6-tetramethyl-1-piperidone, 5000m1 sodium hydroxide solution with a pH value of 12.5, and 18.6g (0.08moL) trichloroiminocyanuric acid one by one while stirring; after reacting at 30 C
under stirring for 3 hours, stand to layer the mixture and separate the organic layer, evaporate the methylene dichloride under vacuum so as to yield 45.Og crude betulinic aldehyde (III); dissolve crude betulinic aldehyde into 450m1 dioxane, add 50ml sodium hydroxide solution with a mass concentration of 30%, and then add 24g silver oxide freshly prepared into the mixture; after reacting at 40 C under stirring for 30 hours, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 20%) into the filtrate until the pH value is 2.5, and then pour the mixture into 5000m1 ice water, extract with methylene dichloride and concentrate so as to provide crude betulinic acid; and recrystallize crude betulinic acid with ethanol so as to yield 23.3g betulinic acid (II) with a purity of 94.0% and yield of 48%.
Embodiment 4 Dissolve 22.2g (0.05moL) betulin (I) into 4440m1 ethyl acetate, and add 0.30g (0.0019 moL) 2,2,6,6-tetramethyl-1-piperidone, 4440ml sodium carbonate solution with a pH value of 9.5, and 8g (0.06moL) N-chlorosuccinimid one by one while stirring; reacting at 75 C under stirring for 12 hours, stand to layer the mixture and separate the organic layer, evaporate ethyl acetate under vacuum so as to yield 25.0 g crude betulinic aldehyde (III);
dissolve the crude betulinic aldehyde into 150m1 acetone, add 30ml sodium hydroxide solution with a mass concentration of 30%, and then add 13g silver oxide prepared freshly into the mixture; after reacting at 25 C under stirring for 72 hours, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 36%) into the filtrate until the pH value is 2.5, and then pour the mixture into 2500ml ice water, extract with methylene dichloride and concentrate so as to provide crude betulinic acid; and recrystallize crude betulinic acid with acetic acid so as to yield 12.1 g betulinic acid (11) with a purity of 92.1 % and yield of 49%.
Embodiment 5 Dissolve 44.3g (0.1 moL) betulin (I) into 5000m1 methylene dichloride, and add 0.35g (0.0022 moL) 2,2,6,6-tetramethyl-1-piperidone, 5000m1 sodium hydroxide solution with a pH value of 12.0, and 26g (0.11 moL) trichloroiminocyanuric acid one by one while stirring; after reacting at 35 C
under stirring for 2 hours, stand to layer the mixture and separate the organic layer, evaporate methylene dichloride under vacuum so as to yield 44.8g crude betulinic aldehyde (III); dissolve crude betulinic aldehyde into 900m1 ethanol, add 50ml sodium hydroxide solution with a mass concentration of 30%, and then add 24g freshly prepared silver oxide into the mixture; after reacting at 75 C for 3 hours under stirring, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 25%) into the filtrate until the pH value is 2.5, and then pour the mixture into 5000ml ice water, extract with methylene dichloride and concentrate so as to provide crude betulinic acid; and recrystallize crude betulinic acid with ethanol so as to yield 23.7g betulinic acid (II) with a purity of 91.2% and yield of 47%.
Embodiment 6 Dissolve 44.3g (0.1 moL) betulin (I) into 5000m1 chloroform, and add 0.16g (0.001moL) 2,2,6,6-tetramethyl -1-piperidone, 5000m1 sodium hydroxide solution with a pH value of 12.0, and 26g (0.11 moL) trichloroiminocyanuric acid one by one under stirring; after reacting at 50 C under stirring for 2 hours, stand to layer the mixture and separate the organic layer, evaporate chloroform under vacuum so as to yield 44.Og crude betulinic aldehyde (III);
dissolve crude betulinic aldehyde into 250m1 propanetriol, add 50ml sodium hydroxide solution with a mass concentration of 10%, and then add 24g silver oxide into the mixture; after reacting at 20 C for 3 hours under stirring, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 36%) into the filtrate until the pH
value is 2.5, and then pour the mixture into 5000ml ice water; keep the mixture standing for 24 hours at 0-5 C, and then filter the mixture so as to provide the crude betulinic acid; and recrystallize crude betulinic acid with ethanol so as to yield 22.6g betulinic acid (II) with a purity of 91.0% and yield of 45%.
1. The present invention is to generate crude betulinic aldehyde by oxidizing betulin and then oxidize crude betulinic aldehyde, so that, in the oxidation product, only bioactive betulinic acid is generated, without non-bioactive a-betulinic acid.
2. The present invention adopts a step-by-step oxidization and highly-selective oxidizer which has no effect on C-3 hydroxyl group, thus functional group protection is not necessary.
3. The present invention has the superiorities of short routing, simple processing method, easily purified product and low production cost, thus industrialized production and popularization is easily achieved.
DESCRIPTION OF DRAWINGS
Fig.1 Schematic diagram for synthesizing betulinic acid through oxidization of betulin DETAILED DESCRIPTION OF THE INVENTION
Further details of the invention are described by combining the Examples of the present invention, as follows:
A method for producing betulinic acid, with the betulin as the raw material, comprising the following steps:
(1) Preparation of crude betulinic aldehyde through oxidation of betulin:
dissolve betulin into an organic solvent, and add 2,2,6,6-tetramethyl-1-piperidone, inorganic alkaline solution with a pH value of 8.012.5 and oxidizer into the mixture; stir to make the mixture react at 2575 C for 0.5-12 hours; upon completion of reaction, keep the mixture standing to layer, separate the organic layer and make the organic phase concentrated so as to provide the grey white crude betulinic aldehyde; the molar ratio of betulin and 2,2,6,6-tetramethyl-1-piperidone is 1:0.01-0.05, and the molar ratio of betulin and the oxidizer is 1: 0.8-1.5; the organic solvent dosage is 100-200ml per gram of betulin; and the inorganic alkaline solution dosage is 100-200ml per gram of betulin;
(2) Preparation of crude betulinic acid through oxidation of crude betulinic aldehyde: dissolve crude betulinic aldehyde generated in step (1) into an organic solvent, and add sodium hydroxide solution and the freshly prepared silver oxide into the mixture; after stirring to make the mixture react at for 3-72 hours, make the reactant cool to room temperature, and filter the reactant; The filtrate is generated betulinic acid through acidification, filtering, washing and drying; the organic solvent dosage is 5-20m1 per gram of crude betulinic aldehyde;
(3) Preparation of betulinic acid through recrystallization of crude betulinic acid: recrystallizing the crude betulinic acid obtained in step (2) by using ethanol, methanol or acetic acid as the solvent to obtain betulinic acid.
The method for producing betulinic acid, wherein, the organic solvent mentioned in step (1) is methylene dichloride, chloroform or ethyl acetate.
The method for producing betulinic acid, wherein, the inorganic alkaline solution mentioned in step (1) is sodium hydroxide solution, sodium phosphate, sodium carbonate solution or sodium bicarbonate solution.
The method for producing betulinic acid, wherein, the oxidizer mentioned in step (1) is trichloroiminocyanuric acid or N-chlorosuccinimide.
The method for producing betulinic acid, wherein, the organic solvent mentioned in step (2) is tetrahydrofuran, dioxane, ethanol or acetone.
Embodiment 1 Dissolve 44.3g (0.1 moL) betulin (I) into 5000m1 methylene dichloride, and add 0.156g (0.001 moL) 2,2,6,6-tetramethyl-1-piperidone, 5000m1 sodium bicarbonate solution with a pH value of 8.3, and 20g (0.15moL) N-chlorosuccinimide one by one while stirring; after reacting at 25 C for 10 hours under stirring, stand to layer the mixture, separate the organic layer, and evaporate methylene dichloride under vacuum so as to yield 45.5g crude betulinic aldehyde (III); dissolve the obtained crude betulinic aldehyde into 910m1 ethanol, add 50ml sodium hydroxide solution with a mass concentration of 30%, and then add 24g freshly prepared silver oxide into the mixture; after react at 70 C under stirring for 5 hours, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 36%) into the filtrate until the pH value is 2.5, and then pour the mixture into 5000m1 ice water, extract with methylene dichloride and concentrate so as to provide crude betulinic acid; and recrystallize crude betulinic acid with methanol so as to yield 23.6g betulinic acid (II) with a purity of 92.9% and yield of 48%.
Betulinic aldehyde is white crystal (methanol); melting point: 190-192 C
(192-193 C); El-MS m/z: 440([M+]), 207, 189, 135; 'H-NMR(300MHz , CDCI3);60.76 (s, 3H, 23-Me) , 0.83(s, 3H, 24-Me), 0.92 (s, 3H, 25-Me), 0.97(s, 3H, 26-Me), 0.98(s,3H,27-Me),1.70(s, 3H, 30-Me),3.19 (q, 3-H), 4.76 (s, 29a-H), 4.63(s, 29b-H), 9.68(s, 28-H);13C-NMR(300MHz, CDCI3038.9(C-1), 28.0(C-2), 79.0(C-3), 40.8(C-4), 59.4(C-5), 18.3(C-6), 34.3(C-7), 40.8(C-8), 50.5(C-9), 37.2(C-10), 20.8(C-11), 25.6(C-12), 38.7(C-13),42.6 (C-14), 29.9 (C-15), 34.3 (C-16), 55.3 (C-17), 47.6 (C-18), 48.1 (C-19), 149.8(C-20), 29.9(C-21), 38.7(C-22), 27.4 (C-23), 15.9(C-24), 16.2(C-25), 16.2(C-26), 14.3(C-27), 206.7(C-28), 110.2 (C-29), 19.0 (C-30).
The betulinic acid is white crystal (methanol); melting point:
284-287 C(285-287 C); El-MS m/z: 456([M+]), 248, 220, 207, 189, 175;
1H-NMR(300MHz,CDCI3) :O0.76(s, 3H, 23-Me), 0.83(s, 3H, 24-Me), 0.94(s, 3H, 25-Me), 0.97(s, 3H, 26-Me), 0.98(s, 3H, 27-Me), 1.69(s, 3H, 30-Me), 3.17(q, 3-H), 4.74(s, 29a-H), 4.62(s, 29b-H); 13C-NMR(300MHz, CDC13): 638.4(C-1), 28.0(C-2), 79.0(C-3), 40.7(C-4), 55.4(C-5), 18.3(C-6), 34.3(C-7), 38.4(C-8), 50.5(C-9), 37.0(C-10), 19.3(C-11), 27.4(C-12), 38.7(C-13), 42.5(C-14), 29.7(C-15), 34.3(C-16), 56.3(C-17), 49.3(C-18), 46.9(C-19), 150.4(C-20), 29.7(C-21), 37.2(C-22), 28.0(C-23), 15.4(C-24), 16.0(C-25), 16.1(C-26), 14.7(C-27), 180.7(C-28),109.7(C-29), 19.4(C-30).
Embodiment 2 Dissolve 44.3g (0.1 moL) betulin (I) into 4430ml chloroform, and add 0.78g (0.005 moL) 2,2,6,6-tetramethyl-1-piperidone, 4430m1 sodium phosphate solution with a pH value of 11.5, and 26g (0.11 moL) trichloroiminocyanuric acid one by one while stirring; after reacting at 50 C for 0.5 hours under stirring, stand to layer the mixture and separate the organic layer, evaporate chloroform under vacuum so as to yield 44.Og crude betulinic aldehyde (III).
Dissolve crude betulinic aldehyde into 220m1 tetrahydrofuran, add 50m1 sodium hydroxide solution with a mass concentration of 30%, and then add 24g freshly prepared silver oxide into the mixture; after reacting at 40 C
under stirring for 30 hours, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 30%) into the filtrate until the pH value is 2.5, and then pour the mixture into 5000m1 ice water, extract with methylene dichloride and concentrate so as to provide crude betulinic acid; and recrystallize crude betulinic acid with ethanol so as to yield 22.6g betulinic acid (II) with a purity of 93.1% and yield of 46%.
Embodiment 3 Dissolve 44.3g (0.1 moL) betulin (I) into 5000m1 methylene dichloride, and add 0.25g (0.0016 moL) 2,2,6,6-tetramethyl-1-piperidone, 5000m1 sodium hydroxide solution with a pH value of 12.5, and 18.6g (0.08moL) trichloroiminocyanuric acid one by one while stirring; after reacting at 30 C
under stirring for 3 hours, stand to layer the mixture and separate the organic layer, evaporate the methylene dichloride under vacuum so as to yield 45.Og crude betulinic aldehyde (III); dissolve crude betulinic aldehyde into 450m1 dioxane, add 50ml sodium hydroxide solution with a mass concentration of 30%, and then add 24g silver oxide freshly prepared into the mixture; after reacting at 40 C under stirring for 30 hours, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 20%) into the filtrate until the pH value is 2.5, and then pour the mixture into 5000m1 ice water, extract with methylene dichloride and concentrate so as to provide crude betulinic acid; and recrystallize crude betulinic acid with ethanol so as to yield 23.3g betulinic acid (II) with a purity of 94.0% and yield of 48%.
Embodiment 4 Dissolve 22.2g (0.05moL) betulin (I) into 4440m1 ethyl acetate, and add 0.30g (0.0019 moL) 2,2,6,6-tetramethyl-1-piperidone, 4440ml sodium carbonate solution with a pH value of 9.5, and 8g (0.06moL) N-chlorosuccinimid one by one while stirring; reacting at 75 C under stirring for 12 hours, stand to layer the mixture and separate the organic layer, evaporate ethyl acetate under vacuum so as to yield 25.0 g crude betulinic aldehyde (III);
dissolve the crude betulinic aldehyde into 150m1 acetone, add 30ml sodium hydroxide solution with a mass concentration of 30%, and then add 13g silver oxide prepared freshly into the mixture; after reacting at 25 C under stirring for 72 hours, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 36%) into the filtrate until the pH value is 2.5, and then pour the mixture into 2500ml ice water, extract with methylene dichloride and concentrate so as to provide crude betulinic acid; and recrystallize crude betulinic acid with acetic acid so as to yield 12.1 g betulinic acid (11) with a purity of 92.1 % and yield of 49%.
Embodiment 5 Dissolve 44.3g (0.1 moL) betulin (I) into 5000m1 methylene dichloride, and add 0.35g (0.0022 moL) 2,2,6,6-tetramethyl-1-piperidone, 5000m1 sodium hydroxide solution with a pH value of 12.0, and 26g (0.11 moL) trichloroiminocyanuric acid one by one while stirring; after reacting at 35 C
under stirring for 2 hours, stand to layer the mixture and separate the organic layer, evaporate methylene dichloride under vacuum so as to yield 44.8g crude betulinic aldehyde (III); dissolve crude betulinic aldehyde into 900m1 ethanol, add 50ml sodium hydroxide solution with a mass concentration of 30%, and then add 24g freshly prepared silver oxide into the mixture; after reacting at 75 C for 3 hours under stirring, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 25%) into the filtrate until the pH value is 2.5, and then pour the mixture into 5000ml ice water, extract with methylene dichloride and concentrate so as to provide crude betulinic acid; and recrystallize crude betulinic acid with ethanol so as to yield 23.7g betulinic acid (II) with a purity of 91.2% and yield of 47%.
Embodiment 6 Dissolve 44.3g (0.1 moL) betulin (I) into 5000m1 chloroform, and add 0.16g (0.001moL) 2,2,6,6-tetramethyl -1-piperidone, 5000m1 sodium hydroxide solution with a pH value of 12.0, and 26g (0.11 moL) trichloroiminocyanuric acid one by one under stirring; after reacting at 50 C under stirring for 2 hours, stand to layer the mixture and separate the organic layer, evaporate chloroform under vacuum so as to yield 44.Og crude betulinic aldehyde (III);
dissolve crude betulinic aldehyde into 250m1 propanetriol, add 50ml sodium hydroxide solution with a mass concentration of 10%, and then add 24g silver oxide into the mixture; after reacting at 20 C for 3 hours under stirring, filter the mixture, acidify the mixture through dropwise adding concentrated hydrochloric acid (mass concentration of 36%) into the filtrate until the pH
value is 2.5, and then pour the mixture into 5000ml ice water; keep the mixture standing for 24 hours at 0-5 C, and then filter the mixture so as to provide the crude betulinic acid; and recrystallize crude betulinic acid with ethanol so as to yield 22.6g betulinic acid (II) with a purity of 91.0% and yield of 45%.
Claims (10)
1. A method for synthesizing betulinic acid, characterized in that betulin will undergo twice selective oxidation; wherein, the oxidation reaction comprises: oxidizing betulin into betulinic aldehyde; then oxidizing betulinic aldehyde into crude betulinic acid; and finally recrystallizing crude betulinic acid to yield betulinic acid.
2. A method for synthesizing betulinic acid, comprising the following steps:
(1)Preparation of betulinic aldehyde by oxidizing betulin:
Firstly, dissolve betulin into an organic solvent, and add 2,2,6,6-tetramethyl-1- piperidone, alkaline solution and oxidizer into the mixture one by one under stirring; then react at 25~75°C for 0.5~12 hours under stirring, so as to prepare betulinic aldehyde;
(2)Preparation of betulinic acid through the oxidation of betulinic aldehyde:
Firstly, dissolve betulinic aldehyde into an organic solvent, and add sodium hydroxide solution and silver oxide into the mixture one by one; then react at 25~85°C for 3-72 hours under stirring, so as to prepare crude betulinic acid through acidification.
(1)Preparation of betulinic aldehyde by oxidizing betulin:
Firstly, dissolve betulin into an organic solvent, and add 2,2,6,6-tetramethyl-1- piperidone, alkaline solution and oxidizer into the mixture one by one under stirring; then react at 25~75°C for 0.5~12 hours under stirring, so as to prepare betulinic aldehyde;
(2)Preparation of betulinic acid through the oxidation of betulinic aldehyde:
Firstly, dissolve betulinic aldehyde into an organic solvent, and add sodium hydroxide solution and silver oxide into the mixture one by one; then react at 25~85°C for 3-72 hours under stirring, so as to prepare crude betulinic acid through acidification.
3. A producing method as claimed in claim 2, characterized in that the molar ratio of betulin and 2,2,6,6-tetramethyl-1-piperidone mentioned in step (1) is 1:0.01~0.05; and the molar ratio of betulin and the oxidizer is 1: 0.8~1.5;
the organic solvent dosage is 100-200ml per gram of betulin; and the alkaline solution dosage is 100-200ml per gram of betulin.
the organic solvent dosage is 100-200ml per gram of betulin; and the alkaline solution dosage is 100-200ml per gram of betulin.
4. A producing method as claimed in claim 2 or 3, characterized in that the organic solvent is selected from one of methylene dichloride, chloroform or ethyl acetate; the alkaline solution is selected from one of sodium hydroxide solution, sodium carbonate solution or sodium bicarbonate solution; the pH
value of the alkaline solution is 8.3~12.5; and the oxidizer is trichloroiminocyanuric acid or N-chlorosuccinimide.
value of the alkaline solution is 8.3~12.5; and the oxidizer is trichloroiminocyanuric acid or N-chlorosuccinimide.
5. A producing method as claimed in claim 2 or 3, characterized in that molar ratio of the betulinic aldehyde and silver oxide mentioned in the step (2) is 1:0.8~1.5.
6. A producing method as claimed in claim 2 or 3, characterized in that the dosage of the organic solvent mentioned in the step (2) is 5~20ml per gram of betulinic aidehyde; the sodium hydroxide solution dosage is 0.5 ~ 1.5 ml per gram of betulinic aldehyde.
7. A producing method as claimed in claim 2 or 3, characterized in that the organic solvent mentioned in the step (2) is selected from one of tetrahydrofuran, dioxane, ethanol or acetone; and the mass concentration of the sodium hydroxide solution is 10-40%.
8. A producing method as claimed in claim 2 or 3, characterized in that the step (1) also comprises: standing to layer the reactant, separating the organic layer and then evaporate the organic layer by vacuum so as to prepare betulinic aldehyde.
9. A producing method as claimed in claim 2 or 3, characterized in that the step (2) also comprises: extracting crude betulinic acid generated in the step (2) and concentrating the extract first, and then recrystallizing so as to yield betulinic acid.
10. A producing method as claimed in claim 9, characterized in that concentrated hydrochloric acid is adopted for the acidification, and the pH
value of the acidulated solution is 2.0~3.0; methylene dichloride is adopted for the extraction; and one of ethanol, methanol or acetic acid is adopted for the recrystallization.
value of the acidulated solution is 2.0~3.0; methylene dichloride is adopted for the extraction; and one of ethanol, methanol or acetic acid is adopted for the recrystallization.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2008100646757A CN101302244B (en) | 2008-06-06 | 2008-06-06 | Production process of betulinic acid |
CN200810064675.7 | 2008-06-06 | ||
PCT/CN2009/071271 WO2009146619A1 (en) | 2008-06-06 | 2009-04-15 | A method for producing betulinic acid |
Publications (2)
Publication Number | Publication Date |
---|---|
CA2705902A1 true CA2705902A1 (en) | 2009-12-10 |
CA2705902C CA2705902C (en) | 2013-05-14 |
Family
ID=40112358
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA2705902A Expired - Fee Related CA2705902C (en) | 2008-06-06 | 2009-04-15 | A method for producing betulinic acid |
Country Status (3)
Country | Link |
---|---|
CN (1) | CN101302244B (en) |
CA (1) | CA2705902C (en) |
WO (1) | WO2009146619A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2755990A1 (en) * | 2011-09-12 | 2014-07-23 | Stora Enso Oyj | Method for preparation of betulinic acid |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101302244B (en) * | 2008-06-06 | 2011-04-13 | 东北林业大学 | Production process of betulinic acid |
CN102344482A (en) * | 2011-07-29 | 2012-02-08 | 温州大学 | Betulinol derivant, preparation method and usage |
SE536995C2 (en) * | 2011-09-12 | 2014-11-25 | Stora Enso Oyj | Procedure for derivatization of a chemical component in wood |
CN103626827A (en) * | 2012-08-29 | 2014-03-12 | 奇复康药物研发(苏州)有限公司 | Preparation method for betulinic acid standard substance |
CN112811995A (en) * | 2021-01-14 | 2021-05-18 | 惠泽化学科技(濮阳)有限公司 | Synthesis method of 4-substituent cyclohexanone liquid crystal intermediate |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5804575A (en) * | 1997-03-27 | 1998-09-08 | The Board Of Trustees Of The University Of Illinois | Methods of manufacturing betulinic acid |
US6232481B1 (en) * | 2000-01-11 | 2001-05-15 | Regents Of The University Of Minnesota | Method for manufacturing betulinic acid |
WO2006105354A1 (en) * | 2005-03-29 | 2006-10-05 | Regents Of The University Of Minnesota | Selective oxidation of triterpenes employing tempo |
CN101302244B (en) * | 2008-06-06 | 2011-04-13 | 东北林业大学 | Production process of betulinic acid |
-
2008
- 2008-06-06 CN CN2008100646757A patent/CN101302244B/en not_active Expired - Fee Related
-
2009
- 2009-04-15 WO PCT/CN2009/071271 patent/WO2009146619A1/en active Application Filing
- 2009-04-15 CA CA2705902A patent/CA2705902C/en not_active Expired - Fee Related
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2755990A1 (en) * | 2011-09-12 | 2014-07-23 | Stora Enso Oyj | Method for preparation of betulinic acid |
EP2755990A4 (en) * | 2011-09-12 | 2015-03-25 | Stora Enso Oyj | Method for preparation of betulinic acid |
Also Published As
Publication number | Publication date |
---|---|
CA2705902C (en) | 2013-05-14 |
CN101302244A (en) | 2008-11-12 |
WO2009146619A1 (en) | 2009-12-10 |
CN101302244B (en) | 2011-04-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CA2705902C (en) | A method for producing betulinic acid | |
CN103724396A (en) | Preparation method of R-budesonide | |
CN113943336A (en) | Method for synthesizing cholesterol by taking BA as raw material | |
Lian-niang et al. | Isolation and Structure Elucidation of Seco-neokadsuranic Acid A and 3, 4-Seco-(24Z)-lanosta-4 (30), 8, 24-triene-3, 26-dioic Acid1 | |
CN102558280A (en) | Method for producing 30-halogenated betulinic acid | |
EP3744711B1 (en) | Method for preparing mesaconine and related intermediaries | |
NO130398B (en) | ||
Singh et al. | Structure and stereochemistry of paristerone, a novel phytoecdysone from the tubers of Paris polyphylla | |
WO2021005618A1 (en) | Synthesis of cholesterol and vitamin d3 from phytosterols | |
CN115716862A (en) | Novel long-acting brassinolide, synthesis method and application thereof | |
Pérez Gil et al. | Synthesis of analogues of brassinosteroids from chenodeoxycholic acid | |
Sharma et al. | Optimization of lantadenes isolation and preparation of 22 β-hydroxyoleanonic acid | |
Sakano et al. | Microbial Conversion of 18 β-Glycyrrhetinic Acid and 22 α-Hydroxy-18 β-glycyrrhetinic Acid by Chainia antibiotic | |
CN102399255B (en) | Preparation method for betulinic acid | |
CN107814824B (en) | Preparation method of tetraene acetate | |
CN111704645A (en) | Application of descimidine reagent in synthesis of Ocotillol type saponin derivative key intermediate | |
Migliuolo et al. | New Δ8-and Δ8 (14)-5α-6α-epoxysterols from the marine sponge Spongia officinalis | |
Gómez-Calvario et al. | Synthetic pathway to 22, 23-dioxocholestanic chain derivatives and their usefulness for obtaining brassinosteroid analogues | |
Symon et al. | Epimerization of hydroxyl group in lupan series triterpenoids | |
RU2397177C2 (en) | METHOD OF PRODUCING 2-DEHYDRO-3-EPI-20-HYDROXYECDYSONE, MINOR ECDYSTEROIDS OF Froelichia floridana SEEDS | |
Kuhl et al. | Partial synthesis of a marine secosterol from Gersemia fruticosa: Preparation of the intermediate precursor 3β, 6α-diacetoxy-24-methyl-12-oxo-5α-chol-9, 11-en-24-oate | |
US3282962A (en) | (optionally 17-alkylated)-17-oxygenated-3-oxa-5alpha-androstan-2-ones and intermediates thereto | |
Liu et al. | Short and Efficient Approach Towards Buxozine‐C, an Alkaloid from Buxus sempervirens | |
González et al. | A new friedooleane triterpenic acid from Schaefferia cuneifolia | |
Bolt | O‐hetero‐analogues of steroids |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request | ||
MKLA | Lapsed |
Effective date: 20220419 |