A kind of preparation method of isoforskolin
Technical field
The present invention relates to the preparation method of isoforskolin (Isoforskolin), more particularly to a kind of different Buddhist department of high-purity
The preparation method of Kelin (Isoforskolin).
Background technology
Studies have shown that labiate Coleus forskohlii (Coleus forskohlii (Wild.) Briq) is derived from, production
In Huize, yunnan, Dongchuan of the country such as India, Nepal and China and other places, with isoforskolin (Isoforskolin) for generation
The labdane diterpenes compound of table, to relevant diseases such as treatment bronchial asthma, congestive heart failure, kinds of tumors
With apparent effect, demand dramatically increases.It is domestic active ingredient purification, expand planting scale, improve effectively at
Point content etc. is done a lot of work.The Chinese patent literature of Patent No. 201110458818.4 discloses a kind of extraction hair
The method that larynx sheath stamen spends active ingredient isoforskolin (Isoforskolin), after Coleus forskohlii herb or root are crushed, with
Hydrous ethanol extracts, and the methods of macroporous resin column chromatography, octadecyl silane column chromatography, recrystallization are purified, and are made
Purity is more than 90% isoforskolin (Isoforskolin).Researcher Hubei Province Tongcheng County manually standardize base plant
Coleus forskohlin medicinal material.Medicinal material is planted compared with Yunnan Wild medicinal material in Tongcheng County, the equal nothing of the indexs such as pharmacognosy, chemical composition and pharmacology
Significant difference, but the content of active ingredient isoforskolin is relatively low.The Chinese patent literature of Patent No. 201310636981.4
A kind of method improving isoforskolin (Isoforskolin) content in coleus forskohlin medicinal material has been disclosed, ultraviolet induction is utilized
Coleus forskohlin medicinal material promotes the method that isoforskolin (Isoforskolin) is synthesized and accumulated.
Due to being limited by suitable cultivation condition, principle active component isoforskolin (Isoforskolin) cannot be satisfied increasingly
The demand (isoforskolin (ISOF) content is only 0.5-1.15 ‰ in dry medicinal material) of growth, different Buddhist department is prepared using chemical synthesis
(Isoforskolin) is imperative for Kelin.
Invention content
The present invention, using Forskolin as raw material, selects suitable solvent in view of the shortcomings of the prior art, using chemical synthesis
System resets chemical synthesis process using base catalysis acetyl group, isolates and purifies, prepare the isoforskolin of purity >=99%
(Isoforskolin), meets the needs of isoforskolin is growing, industrialized production is significant.
Technical scheme is as follows:
A kind of preparation method of isoforskolin, using Forskolin as raw material, using basic catalyst, in methylene chloride
Isoforskolin is obtained by acetyl group rearrangement reaction.
Preferably, the preparation method of above-mentioned isoforskolin, the basic catalyst is alkalescent catalyst, selected from alkalinity
One or more of aluminium oxide, basic resin, alkaline silica gel or alkaline diatomite.The grain size of further preferred alkali alumina
For 200-400 mesh, pH value 9.0-10.0 (100g/L, 25 DEG C).
Basic resin can be weakly base resin, such as 717 type ion exchange resin, D301 macroreticular resins.
The preparation method of above-mentioned isoforskolin, preferably Forskolin:Alkali alumina:Dichloromethane=1g:(10-
30)g:(10-100)ml.More preferable Forskolin:Alkali alumina:Dichloromethane=1g:20g:25ml.
The preparation method of above-mentioned isoforskolin, preferable reaction temperature are 15 DEG C -40 DEG C, reaction time 48h-72h.
The preparation method of above-mentioned isoforskolin can further include the separating step to isoforskolin.It can be with
By the reaction solution filtering containing isoforskolin, then soaked in solvent washing catalyst, is distilled after merging with filtrate and obtain different Buddhist
Scoline crude product is, it is preferable to use methanol washing by soaking catalyst, further preferred catalyst:Cleaning solvent=1kg:1.5L.
The preparation method of isoforskolin of the present invention can also further comprise the purifying to isoforskolin
Step.It is preferred that being recrystallized to isoforskolin crude product.
One preferred purification schemes is:It is purified using secondary crystallization, first time crystallization purifying, solvent burden ratio (volume ratio)
For ethyl acetate (EA):Petroleum ether (PE)=1:2, solvent dosage is crude product (kg):Recrystallisation solvent (L)=1:9, it is heated to back
Stream, maintains the reflux for 2h, is naturally cooling to 25 DEG C, and filtering is crystallized (I) filter cake EA:PE=1:The solvent of 3 (about 200ml)
It washs (3-5 times);Second of crystallization purifying, solvent burden ratio (volume ratio) are ethyl acetate (EA):Petroleum ether (PE)=1:2, it is molten
Agent dosage is crystallization (I) (kg):Recrystallisation solvent (L)=1:9, it is heated to flowing back, maintains the reflux for 2h, be naturally cooling to 25 DEG C, mistake
Filter, is prepared the high-purity isoforskolin (Isoforskolin) of purity >=99%, filter cake EA:PE=1:3 (about
Solvent washing (3-5 times) 200ml);Solvent recovery cycle in mother liquor uses, spare after solid constituent analysis.
Beneficial effects of the present invention are:
(1) single step reaction is used, target product selectivity is good, and reaction system is cleaner, and by-product is less, and the reaction used time is very
It is short;
(2) select solvent toxicity smaller;
(3) crystallization yield is higher, the recyclable application of solvent, can reach 99% or more purity after crystallizing twice.
In conclusion the present invention requires catalyst, solvent, reaction condition etc. low, it is a kind of suitable industrialized production
Isoforskolin new technology paths, have the used time it is short, high income is of low cost, easy to operate, it is safe the advantages that.
Description of the drawings
Fig. 1 is the HPLC figures for the isoforskolin that the method for the invention is prepared.
Fig. 2 is the LCMS figures for the isoforskolin that the method for the invention is prepared.
Fig. 3 is the NMR figures for the isoforskolin that the method for the invention is prepared.
Specific implementation mode
Below by the drawings and specific embodiments, the invention will be further described, but is not meant to protect the present invention
Protect the limitation of range.
The investigation of 1 reaction condition of embodiment
Using Forskolin as raw material, using grain size:200-400 mesh, pH value (100g/L, 25 DEG C):9.0-10.0 alkalinity oxygen
Change Al catalysts, select dichloromethane, tetrahydrofuran, methanol, ethyl alcohol, benzene for reaction dissolvent respectively, investigates under room temperature to different Buddhist
Influence prepared by scoline;Reaction vessel is closed using nitrogen;Using mechanical agitation mode, acetyl group rearrangement reaction is carried out.It is complete
At being filtered after reaction, with catalytic amount (kg):Cleaning solvent (L)=1:1.5 methanol (MeOH) washing by soaking catalyst, filtrate are closed
And distillation obtains crude product afterwards;
Reaction dissolvent |
Response situation |
Reaction time |
Yield |
Dichloromethane |
Reaction is very fast |
There is no product generations, reaction to complete after 48-72h |
53% |
Tetrahydrofuran |
Reaction is very slow |
Reaction only has a small amount of target product to 72h and generates. |
/ |
Methanol |
Reaction is very slow |
Reaction only has a small amount of target product to 72h and generates. |
/ |
Ethyl alcohol |
Reaction is very slow |
Reaction only has a small amount of target product to 72h and generates. |
/ |
Benzene |
Reaction is slower |
There is no product generations, reaction to complete after 120h. |
55% |
By the comparison to different solvents, it is found that reaction dissolvent has a significant impact to preparation method of the present invention, select tetrahydrochysene
As reaction dissolvent, reaction speed is very slow for furans, methanol and ethyl alcohol, and reaction to 72h only has a small amount of target product and generates.Such as
Fruit heats reaction system, then by-product increases much in reaction system.Select benzene and dichloromethane as solvent, the two yield
Quite, but benzene is as reaction dissolvent, and reaction speed is slower, and needing 120h ability, the reaction was complete, and selects dichloromethane, reacts
There are many speed, and 48h can reach that the reaction was complete.Since benzene has prodigious toxicity, and the long time is not suitable for
Industrialized production, therefore the reaction dissolvent for selecting dichloromethane to be prepared as isoforskolin.
Embodiment 2
A kind of preparation method of isoforskolin (Isoforskolin), specifically includes following steps described in the present embodiment:
(1) using Forskolin as raw material, using grain size:200-400 mesh, pH value (100g/L, 25 DEG C):9.0-10.0 alkaline
Aluminium oxide catalyst, dichloromethane (DCM) solvent;
(2) in suitable condition:1. raw material dosage:Catalyst amount:Solvent dosage=1g:20g:25ml;2. 15 DEG C of temperature;
3. time 48h;4. reaction vessel is closed using nitrogen;5. using mechanical agitation mode, acetyl group rearrangement reaction is carried out;
(3) it completes to filter after 1. reacting, with catalytic amount (kg):Cleaning solvent (L)=1:1.5 methanol (MeOH) immersion is washed
Catalyst is washed, filtrate is distilled after merging obtains crude product;
(4) secondary crystallization is used to purify, first time crystallization purifying, solvent burden ratio (volume ratio) is ethyl acetate (EA):Stone
Oily ether (PE)=1:2, solvent dosage is crude product (kg):Recrystallisation solvent (L)=1:9, it is heated to flowing back, maintains the reflux for 2h, it is natural
25 DEG C are cooled to, filtering is crystallized (I) filter cake EA:PE=1:The solvent washing (3-5 times) of 3 (about 200ml);Second
Crystallization purifying, solvent burden ratio (volume ratio) are ethyl acetate (EA):Petroleum ether (PE)=1:2, solvent dosage is crystallization (I)
(kg):Recrystallisation solvent (L)=1:9, it is heated to flowing back, maintains the reflux for 2h, be naturally cooling to 25 DEG C, purity is prepared in filtering
>=99% high-purity isoforskolin (Isoforskolin), filter cake EA:PE=1:The solvent of 3 (about 200ml) washs (3-
5 times);Solvent recovery cycle in mother liquor uses, spare after solid constituent analysis.Yield 53%.
The results are shown in Figure 1 by the HPLC of the present embodiment.HPLC conditions are as follows:
Column:sun fire(150mm×4.6mm);
Mobile phase:A:H2O+0.1%THA, B:ACN+0.1%THA;
Min |
A (%) |
B (%) |
0 |
95 |
5 |
2 |
95 |
5 |
5 |
70 |
30 |
20 |
25 |
75 |
30 |
5 |
95 |
Isoforskolin retention time is 15.304min, purity 99.3%.
H1NMR(DMSO-d6):
δ=7.69 (s, 1H), 6.22 (d, 1H), 6.10 (dd, 1H), 5.54 (dd, 1H), 5.10 (d, 1H), 4.84 (dd,
1H),4.74(d,1H),4.42(d,1H),4.02(t,1H),3.05(d,1H),2.45(d,1H),2.17(d,1H),2.04(m,
4H),1.69(m,1H),1.45(s,3H),1.29,1.30(m,7H),1.01(m,4H),0.89(s,3H).
LCMS, NMR of the present embodiment are as shown in Figure 2,3.
Embodiment 3
A kind of preparation method of isoforskolin (Isoforskolin), specifically includes following steps described in the present embodiment:
(1) using Forskolin as raw material, using grain size:200-400 mesh, pH value (100g/L, 25 DEG C):9.0-10.0 alkaline
Aluminium oxide catalyst, dichloromethane (DCM) solvent;
(2) in suitable condition:1. raw material dosage:Catalyst amount:Solvent dosage=1g:20g:25ml;2. 25 DEG C of temperature;
3. time 60h;4. reaction vessel is closed using argon gas;5. using mechanical agitation mode, acetyl group rearrangement reaction is carried out;
(3) it completes to filter after 1. reacting, with catalytic amount (kg):Cleaning solvent (L)=1:1.5 methanol (MeOH) immersion is washed
Catalyst is washed, filtrate is distilled after merging obtains crude product;
(4) secondary crystallization is used to purify, first time crystallization purifying, solvent burden ratio (volume ratio) is ethyl acetate (EA):Stone
Oily ether (PE)=1:2, solvent dosage is crude product (kg):Recrystallisation solvent (L)=1:9, it is heated to flowing back, maintains the reflux for 2h, it is natural
25 DEG C are cooled to, filtering is crystallized (I) filter cake EA:PE=1:The solvent washing (3-5 times) of 3 (about 200ml);Second
Crystallization purifying, solvent burden ratio (volume ratio) are ethyl acetate (EA):Petroleum ether (PE)=1:2, solvent dosage is crystallization (I)
(kg):Recrystallisation solvent (L)=1:9, it is heated to flowing back, maintains the reflux for 2h, be naturally cooling to 25 DEG C, purity is prepared in filtering
>=99% high-purity isoforskolin (Isoforskolin), filter cake EA:PE=1:The solvent of 3 (about 200ml) washs (3-
5 times);Solvent recovery cycle in mother liquor uses, spare after solid constituent analysis.Yield 53%.
H1NMR(DMSO-d6):
δ=7.69 (s, 1H), 6.22 (d, 1H), 6.10 (dd, 1H), 5.54 (dd, 1H), 5.10 (d, 1H), 4.84 (dd,
1H),4.74(d,1H),4.42(d,1H),4.02(t,1H),3.05(d,1H),2.45(d,1H),2.17(d,1H),2.04(m,
4H),1.69(m,1H),1.45(s,3H),1.29,1.30(m,7H),1.01(m,4H),0.89(s,3H).
HPLC, LCMS, NMR qualification result of the present embodiment are same as Example 2.
Embodiment 4
A kind of preparation method of isoforskolin (Isoforskolin), specifically includes following steps described in the present embodiment:
(1) using Forskolin as raw material, using grain size:200-400 mesh, pH value (100g/L, 25 DEG C):9.0-10.0 alkaline
Aluminium oxide catalyst, dichloromethane (DCM) solvent;
(2) in suitable condition:1. raw material dosage:Catalyst amount:Solvent dosage=1g:20g:25ml;2. 40 DEG C of temperature;
3. time 72h;4. reaction vessel is closed using nitrogen;5. using mechanical agitation mode, acetyl group rearrangement reaction is carried out;
(3) it completes to filter after 1. reacting, with catalytic amount (kg):Cleaning solvent (L)=1:1.5 methanol (MeOH) immersion is washed
Catalyst is washed, filtrate is distilled after merging obtains crude product;
(4) secondary crystallization is used to purify, first time crystallization purifying, solvent burden ratio (volume ratio) is ethyl acetate (EA):Stone
Oily ether (PE)=1:2, solvent dosage is crude product (kg):Recrystallisation solvent (L)=1:9, it is heated to flowing back, maintains the reflux for 2h, it is natural
25 DEG C are cooled to, filtering is crystallized (I) filter cake EA:PE=1:The solvent washing (3-5 times) of 3 (about 200ml);Second
Crystallization purifying, solvent burden ratio (volume ratio) are ethyl acetate (EA):Petroleum ether (PE)=1:2, solvent dosage is crystallization (I)
(kg):Recrystallisation solvent (L)=1:9, it is heated to flowing back, maintains the reflux for 2h, be naturally cooling to 25 DEG C, purity is prepared in filtering
>=99% high-purity isoforskolin (Isoforskolin), filter cake EA:PE=1:The solvent of 3 (about 200ml) washs (3-
5 times);Solvent recovery cycle in mother liquor uses, spare after solid constituent analysis.Yield 53%.
H1NMR(DMSO-d6):
δ=7.69 (s, 1H), 6.22 (d, 1H), 6.10 (dd, 1H), 5.54 (dd, 1H), 5.10 (d, 1H), 4.84 (dd,
1H),4.74(d,1H),4.42(d,1H),4.02(t,1H),3.05(d,1H),2.45(d,1H),2.17(d,1H),2.04(m,
4H),1.69(m,1H),1.45(s,3H),1.29,1.30(m,7H),1.01(m,4H),0.89(s,3H).
HPLC, LCMS, NMR qualification result of the present embodiment are same as Example 2.