CN104013972A - Production method of hydrophilic betulinic acid preparation - Google Patents
Production method of hydrophilic betulinic acid preparation Download PDFInfo
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- CN104013972A CN104013972A CN201410289470.4A CN201410289470A CN104013972A CN 104013972 A CN104013972 A CN 104013972A CN 201410289470 A CN201410289470 A CN 201410289470A CN 104013972 A CN104013972 A CN 104013972A
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Abstract
The invention provides a production method of a hydrophilic betulinic acid preparation and relates to a preparation method of a betulinic acid preparation. The production method of the hydrophilic betulinic acid preparation aims at solving the problem that the use of the betulinic acid in the aspect of medical preparations is limited because the betulinic acid is insoluble in water. The method comprises the following steps: 1, preparing 6-p-toluenesulphonyl-beta-cyclodextrin, 2, preparing 6-diethylenetriamine chain-beta-cyclodextrin, 3, preparing a diethylenetriamine bridged-beta-cyclodextrin dimer, and 4, weighing 6-diethylenetriamine chain-beta-cyclodextrin or the diethylenetriamine bridged-beta-cyclodextrin dimer and putting the weighed sample in a sample tube, adding distilled water to the sample tube for dissolving the sample, next, adding the betulinic acid, carrying out ultrasonic oscillation, continuing oscillating on an oscillator, centrifuging, filtering the supernatant liquor by use of a water-phase cellulose filter membrane, and then drying the filtrate, thereby obtaining a white powder, namely the hydrophilic betulinic acid preparation. The production method is used for producing the betulinic acid preparation.
Description
Technical field
The present invention relates to a kind of preparation method of belulinic acid Betulinic acid preparation.
Background technology
Belulinic acid Betulinic acid (betulinicacid), has another name called betulic acid, is a kind of compound of pentacyclic triterpene, is extensively present in Cortex Betulae Luminiferae and other plant, also can make by betulinol is synthetic molecular weight 456.71g/mol, colourless acicular crystal, molecular formula: C
30h
48o
3, molecular structural formula:
Belulinic acid Betulinic acid, as natural bioactive substance, has the effects such as malaria, antiinflammatory.In research, find that belulinic acid Betulinic acid has good antitumaous effect to mankind's black cancer, also has good therapeutic effect to diseases such as lymphatic cancer, carcinoma of prostate.Belulinic acid Betulinic acid is because have the effect that antitumor increases, and also having new mechanism of action aspect anti-acquired immune deficiency syndrome (AIDS) (acquired immune deficiency syndrome (AIDS)), caused the extensive concern of research worker.Belulinic acid Betulinic acid not only has many medical mechanisms, but also does not synthesize the toxicity of quasi drugs, does not also synthesize the drug resistance of quasi drugs, so belulinic acid Betulinic acid has DEVELOPMENT PROSPECT widely aspect medicament research and development.
But because belulinic acid Betulinic acid is insoluble in water, limited its application aspect pharmaceutical preparation.
Cyclodextrin (Cyclodextrin is called for short CD) is a kind of cyclic oligomer sugar compounds, has special ring-type hollow cylinder type structure, and internal diameter 0.7～0.8nm can form clathrate with enclosed molecule (as medicine) under certain condition.Conventional beta-schardinger dextrin-has the loop configuration being comprised of 7 glucose molecules,, after suitably processing, pharmaceutical pack can be wrapped in its circulus and form ultra micro capsule clathrate with medicine, can increase dissolubility, the stability of medicine.But this dissolubility in water of beta-schardinger dextrin-is not very large, so the efficiency of enclose medicine is also lower.
Summary of the invention
The present invention will solve belulinic acid Betulinic acid to be insoluble in water, and has limited the problem of its application aspect pharmaceutical preparation, and a kind of preparation method of hydrophilic belulinic acid Betulinic acid preparation is provided.
The preparation method of hydrophilic belulinic acid Betulinic acid preparation of the present invention, carry out according to the following steps:
One, taking 40～60g beta-schardinger dextrin-joins in 400～500mL distilled water, obtain cyclodextrin solution, getting 6～7g NaOH is dissolved in 20～25mL distilled water, obtain NaOH solution, the NaOH solution of preparation is dropwise joined in cyclodextrin solution, obtain transparent and homogeneous solution, getting 7～10g p-methyl benzene sulfonic chloride is dissolved in 30～40mL acetonitrile, obtain mixed solution, mixed solution is dropwise joined in transparent and homogeneous solution, stirring reaction 2～4h at room temperature, sucking filtration is precipitated, filtrate is preserved to 12h at 0～5 ℃, sucking filtration must precipitate again, by the precipitation vacuum drying 12～16h obtaining for twice, obtain white solid, with absolute ethanol washing white solid 3～5 times, each dehydrated alcohol 50～80mL that uses, obtain 6-tolysulfonyl-beta-schardinger dextrin-,
Two, take 1.5～3g6-tolysulfonyl-beta-schardinger dextrin-, 20～30mgKI and 0.5～1.0mL diethylenetriamine are dissolved in the N-Methyl pyrrolidone that 5～8mL is dry, stirring reaction 4～7h under 70～80 ℃ of conditions, obtain yellow solution, be cooled to room temperature, yellow solution is poured in 100～150mL dehydrated alcohol and separated out white precipitate, sucking filtration is collected white precipitate, respectively with 100～150mL dehydrated alcohol and the washing of 50～80mL ether, vacuum drying obtains 1.3～2.5g product A, product A is dissolved with 8～12mL distilled water, with hydrogen ion exchange resin post, carry out eluting, eluant is followed successively by 400～500mL distilled water, the ammonia of 400～500mL1mol/L, the eluent of collecting ammonia part is concentrated, vacuum drying, obtain product B, product B is dissolved in to 20～30mL distilled water, decompression, concentrate drying obtains 6-diethylenetriamine chain-beta-schardinger dextrin-,
Three, take 2.6～3.9g6-tolysulfonyl-beta-schardinger dextrin-and 1.2～2.4g6-diethylenetriamine chain-beta-schardinger dextrin-, be dissolved in 80～100mLN, in dinethylformamide, at N
2under protection and 75～85 ℃ of condition of water bath heating, reaction is stirred 2～3 days, reaction solution is concentrated into 20～30mL, then join in 300～400mL acetone and produce and precipitate, sucking filtration also precipitates 3～5 times with acetone cyclic washing, precipitation is dissolved in 10～15mL distilled water and with water cellulose filter membrane again and filters, filtrate is dried to obtain diethylenetriamine bridging-beta-schardinger dextrin-dimer;
Four, produce clathrate: take 6-diethylenetriamine chain-beta-schardinger dextrin-of 0.5～0.7g or 0.7～1.0g diethylenetriamine bridging-beta-schardinger dextrin-dimer in sample cell, add 5～6mL distilled water to make its dissolving, add again 30～40mg belulinic acid Betulinic acid, after sonic oscillation 20～60min, on agitator, continue vibration 6～7 days, be placed on afterwards in high speed centrifuge with the centrifugal 15～30min of 10000～12000rpm, getting supernatant filters with water cellulose filter membrane, again filtrate is dried, obtain white powder, be hydrophilic belulinic acid Betulinic acid preparation.
Described in step 2, resin column specification is: 25 * 450mm, and 724 weak-acid cation-exchange resins, resin filling height is 250mm.
Beneficial effect of the present invention:
The present invention is by cyclodextrin derivatization, and the cyclodextrin after derivatization not only very big degree has increased dissolubility own, and can also promote the enclose to medicine.Utilize synthetic cyclodextrin derivative to carry out enclose to belulinic acid Betulinic acid, can effectively improve the dissolubility of belulinic acid Betulinic acid in water, thereby solve the difficult problem of belulinic acid Betulinic acid aspect biological preparation.
Accompanying drawing explanation
Fig. 1 is the infrared spectrum of the hydrophilic belulinic acid Betulinic acid preparation of belulinic acid Betulinic acid, 6-diethylenetriamine chain-beta-schardinger dextrin-, the two physical mixture and embodiment 1 preparation; Fig. 2 is the X-ray diffractogram of the hydrophilic belulinic acid Betulinic acid preparation of belulinic acid Betulinic acid, 6-diethylenetriamine chain-beta-schardinger dextrin-, the two physical mixture and embodiment 1 preparation; Fig. 3 is belulinic acid Betulinic acid standard specimen mass spectrum; Fig. 4 is the mass spectrum of the de-thing of contracting for fixed output quotas of hydrophilic belulinic acid Betulinic acid preparation of embodiment 1 preparation; Fig. 5 is belulinic acid Betulinic acid standard specimen UPLC figure; Fig. 6 is the UPLC figure of the de-thing of contracting for fixed output quotas of hydrophilic belulinic acid Betulinic acid preparation of embodiment 1 preparation; Fig. 7 is the infrared spectrum of the hydrophilic belulinic acid Betulinic acid preparation of belulinic acid Betulinic acid, diethylenetriamine bridging beta-schardinger dextrin-dimer, the two physical mixture and embodiment 2 preparations; Fig. 8 is the X-ray diffractogram of the hydrophilic belulinic acid Betulinic acid preparation of belulinic acid Betulinic acid, diethylenetriamine bridging beta-schardinger dextrin-dimer, the two physical mixture and embodiment 2 preparations; Fig. 9 is the mass spectrum of the de-thing of contracting for fixed output quotas of hydrophilic belulinic acid Betulinic acid preparation of embodiment 2 preparations; Figure 10 is the UPLC figure of the de-thing of contracting for fixed output quotas of the hydrophilic belulinic acid Betulinic acid preparation prepared of embodiment.
The specific embodiment
Technical solution of the present invention is not limited to the following cited specific embodiment, also comprises the combination in any between each specific embodiment.
The specific embodiment one: the preparation method of present embodiment hydrophilic belulinic acid Betulinic acid preparation, carry out according to the following steps:
One, taking 40～60g beta-schardinger dextrin-joins in 400～500mL distilled water, obtain cyclodextrin solution, getting 6～7g NaOH is dissolved in 20～25mL distilled water, obtain NaOH solution, the NaOH solution of preparation is dropwise joined in cyclodextrin solution, obtain transparent and homogeneous solution, getting 7～10g p-methyl benzene sulfonic chloride is dissolved in 30～40mL acetonitrile, obtain mixed solution, mixed solution is dropwise joined in transparent and homogeneous solution, stirring reaction 2～4h at room temperature, sucking filtration is precipitated, filtrate is preserved to 12h at 0～5 ℃, sucking filtration must precipitate again, by the precipitation vacuum drying 12～16h obtaining for twice, obtain white solid, with absolute ethanol washing white solid 3～5 times, each dehydrated alcohol 50～80mL that uses, obtain 6-tolysulfonyl-beta-schardinger dextrin-,
Two, take 1.5～3g6-tolysulfonyl-beta-schardinger dextrin-, 20～30mgKI and 0.5～1.0mL diethylenetriamine are dissolved in the N-Methyl pyrrolidone that 5～8mL is dry, stirring reaction 4～7h under 70～80 ℃ of conditions, obtain yellow solution, be cooled to room temperature, yellow solution is poured in 100～150mL dehydrated alcohol and separated out white precipitate, sucking filtration is collected white precipitate, respectively with 100～150mL dehydrated alcohol and the washing of 50～80mL ether, vacuum drying obtains 1.3～2.5g product A, product A is dissolved with 8～12mL distilled water, with hydrogen ion exchange resin post, carry out eluting, eluant is followed successively by 400～500mL distilled water, the ammonia of 400～500mL1mol/L, the eluent of collecting ammonia part is concentrated, vacuum drying, obtain product B, product B is dissolved in to 20～30mL distilled water, decompression, concentrate drying obtains 6-diethylenetriamine chain-beta-schardinger dextrin-,
Three, take 2.6～3.9g6-tolysulfonyl-beta-schardinger dextrin-and 1.2～2.4g6-diethylenetriamine chain-beta-schardinger dextrin-, be dissolved in 80～100mLN, in dinethylformamide, at N
2under protection and 75～85 ℃ of condition of water bath heating, reaction is stirred 2～3 days, reaction solution is concentrated into 20～30mL, then join in 300～400mL acetone and produce and precipitate, sucking filtration also precipitates 3～5 times with acetone cyclic washing, precipitation is dissolved in 10～15mL distilled water and with water cellulose filter membrane again and filters, filtrate is dried to obtain diethylenetriamine bridging-beta-schardinger dextrin-dimer;
Four, produce clathrate: take 6-diethylenetriamine chain-beta-schardinger dextrin-of 0.5～0.7g or 0.7～1.0g diethylenetriamine bridging-beta-schardinger dextrin-dimer in sample cell, add 5～6mL distilled water to make its dissolving, add again 30～40mg belulinic acid Betulinic acid, after sonic oscillation 20～60min, on agitator, continue vibration 6～7 days, be placed on afterwards in high speed centrifuge with the centrifugal 15～30min of 10000～12000rpm, getting supernatant filters with water cellulose filter membrane, again filtrate is dried, obtain white powder, be hydrophilic belulinic acid Betulinic acid preparation.
The specific embodiment two: present embodiment is different from the specific embodiment one: take 2g6-tolysulfonyl-beta-schardinger dextrin-, 25mgKI and 0.57mL diethylenetriamine in step 2 and be dissolved in the N-Methyl pyrrolidone that 5mL is dry.Other is identical with the specific embodiment one.
The specific embodiment three: present embodiment is different from the specific embodiment one or two: described in step 2, resin column specification is: 25 * 450mm, 724 weak-acid cation-exchange resins, resin filling height is 250mm.Other is identical with the specific embodiment one or two.
The specific embodiment four: present embodiment is different from one of specific embodiment one to three: take 2.6g6-tolysulfonyl-beta-schardinger dextrin-and 2.4g6-diethylenetriamine chain-beta-schardinger dextrin-in step 3, be dissolved in 80mLN, in dinethylformamide.Other is identical with one of specific embodiment one to three.
The specific embodiment five: present embodiment is different from one of specific embodiment one to four: take 6-diethylenetriamine chain-beta-schardinger dextrin-of 0.6g or 0.8g diethylenetriamine bridging-beta-schardinger dextrin-dimer in step 4 in sample cell.Other is identical with one of specific embodiment one to four.
The specific embodiment six: present embodiment is different from one of specific embodiment one to five: add 35mg belulinic acid Betulinic acid in step 4.Other is identical with one of specific embodiment one to five.
The specific embodiment seven: present embodiment is different from one of specific embodiment one to six: the dry freeze-drying that adopts described in step 4, be specially: filtrate is placed in freezer dryer, at-50 ℃ of precooling 6～8h, vacuum drying 24～36h under 30～200Pa.Other is identical with one of specific embodiment one to six.
Embodiment 1:
The preparation method of the present embodiment hydrophilic belulinic acid Betulinic acid preparation, carry out according to the following steps:
One, taking 60g beta-schardinger dextrin-joins in 500mL distilled water, obtain cyclodextrin solution, getting 6.5gNaOH is dissolved in 20mL distilled water, obtain NaOH solution, the NaOH solution of preparation is dropwise joined in cyclodextrin solution, obtain transparent and homogeneous solution, getting 10g p-methyl benzene sulfonic chloride is dissolved in 30mL acetonitrile, obtain mixed solution, mixed solution is dropwise joined in transparent and homogeneous solution, stirring reaction 2h at room temperature, sucking filtration is precipitated, filtrate is preserved to 12h at 5 ℃, sucking filtration must precipitate again, by the precipitation vacuum drying 12h obtaining for twice, obtain white solid, with absolute ethanol washing white solid 4 times, each dehydrated alcohol 60mL that uses, obtain 6.4g6-tolysulfonyl-beta-schardinger dextrin-,
Two, take 2g6-tolysulfonyl-beta-schardinger dextrin-, 25mgKI and 0.57mL diethylenetriamine are dissolved in the N-Methyl pyrrolidone that 5mL is dry, stirring reaction 4h under 70 ℃ of conditions, obtain yellow solution, be cooled to room temperature, yellow solution is poured in 100mL dehydrated alcohol and separated out white precipitate, sucking filtration is collected white precipitate, respectively with 100mL dehydrated alcohol and the washing of 50mL ether, vacuum drying obtains 1.7g product A, product A is dissolved with 10mL distilled water, with hydrogen ion exchange resin post, carry out eluting, eluant is followed successively by 400mL distilled water, the ammonia of 500mL1mol/L, the eluent of collecting ammonia part is concentrated, vacuum drying, obtain product B, product B is dissolved in to 20mL distilled water, decompression, concentrate drying obtains 1.4g6-diethylenetriamine chain-beta-schardinger dextrin-,
Three, produce clathrate: take 6-diethylenetriamine chain-beta-schardinger dextrin-of 0.5g in sample cell, add 5mL distilled water to make its dissolving, add again 30mg belulinic acid Betulinic acid, after sonic oscillation 30min, on agitator, continue vibration 7 days, be placed on afterwards in high speed centrifuge with the centrifugal 15min of 10000rpm, getting supernatant filters with water cellulose filter membrane, again filtrate is dried, obtains white powder, be hydrophilic belulinic acid Betulinic acid preparation.
The dry freeze-drying that adopts described in step 3, is specially: filtrate is placed in freezer dryer, and at-50 ℃ of precooling 6h, vacuum drying 24h under 200Pa.
Utilize infrared, mass spectrum, thermogravimetric analysis, X-ray diffraction and nuclear magnetic resonance, NMR to characterize, and the solution after utilizing super effect liquid phase to the de-bag of clathrate carry out the detection of belulinic acid Betulinic acid content.Through super effect liquid phase, recording belulinic acid Betulinic acid content in hydrophilic belulinic acid Betulinic acid preparation prepared by the present embodiment is 329.75 μ g/mL.
Fig. 1 is the infrared spectrum of the hydrophilic belulinic acid Betulinic acid preparation prepared of belulinic acid Betulinic acid, 6-diethylenetriamine chain-beta-schardinger dextrin-, the two physical mixture and the present embodiment.In Fig. 1, a is belulinic acid Betulinic acid, and b is 6-diethylenetriamine chain-beta-schardinger dextrin-, and c is the two physical mixture, and d is hydrophilic belulinic acid Betulinic acid preparation.Can find out that belulinic acid Betulinic acid and 6-diethylenetriamine chain-beta-schardinger dextrin-respectively have the infrared signature absworption peak of self, the spectrogram of physical mixture has additivity, show as the stack of two components, and hydrophilic belulinic acid Betulinic acid preparation is different, the C=O stretching vibration absworption peak (1686.34cm of belulinic acid Betulinic acid association carboxylic acid
-1) and C-O stretching vibration peak (1235.71cm
-1) disappear, show that belulinic acid Betulinic acid, by enclose, has formed clathrate.
Fig. 2 is the X-ray diffractogram of the hydrophilic belulinic acid Betulinic acid preparation prepared of belulinic acid Betulinic acid, 6-diethylenetriamine chain-beta-schardinger dextrin-, the two physical mixture and the present embodiment.In Fig. 2, a is belulinic acid Betulinic acid, and b is 6-diethylenetriamine chain-beta-schardinger dextrin-, and c is the two physical mixture, and d is hydrophilic belulinic acid Betulinic acid preparation.Belulinic acid Betulinic acid and 6-diethylenetriamine chain-beta-schardinger dextrin-have visibly different diffraction maximum separately, in physical mixture spectrogram, the diffraction maximum of existing belulinic acid Betulinic acid has again the diffraction maximum of 6-diethylenetriamine chain-beta-schardinger dextrin-, and hydrophilic belulinic acid Betulinic acid preparation is different from physical mixture, the Partial Feature diffraction maximum of belulinic acid Betulinic acid disappears, show really to have formed new thing phase, formed clathrate.
Fig. 3 is belulinic acid Betulinic acid standard specimen mass spectrum, Fig. 4 is the mass spectrum of the de-thing of contracting for fixed output quotas of the hydrophilic belulinic acid Betulinic acid preparation prepared of the present embodiment, can see the molecular ion peak that in belulinic acid Betulinic acid standard substance figure, stable existence mass-to-charge ratio is 454.8, in the de-thing of contracting for fixed output quotas of hydrophilic belulinic acid Betulinic acid preparation, exist 455.0 molecular ion peak and intensity large, illustrate to take off in the thing of contracting for fixed output quotas and contain belulinic acid Betulinic acid.
Fig. 5 is belulinic acid Betulinic acid standard specimen UPLC figure, Fig. 6 is the UPLC figure of the de-thing of contracting for fixed output quotas of the hydrophilic belulinic acid Betulinic acid preparation prepared of the present embodiment, de-belulinic acid Betulinic acid integration peak area of contracting for fixed output quotas in thing is 74407, be brought in belulinic acid Betulinic acid standard curve linear equation A=6705.25C (μ g/mL)-14037.09 (correlation coefficient 0.9997), obtaining concentration is 13.19ppm, and known belulinic acid Betulinic acid dissolubility in water increases to 329.75 μ g/mL as calculated.
Embodiment 2:
The preparation method of the present embodiment hydrophilic belulinic acid Betulinic acid preparation, carry out according to the following steps:
One, taking 50g beta-schardinger dextrin-joins in 500mL distilled water, obtain cyclodextrin solution, getting 6gNaOH is dissolved in 20mL distilled water, obtain NaOH solution, the NaOH solution of preparation is dropwise joined in cyclodextrin solution, obtain transparent and homogeneous solution, getting 8g p-methyl benzene sulfonic chloride is dissolved in 30mL acetonitrile, obtain mixed solution, mixed solution is dropwise joined in transparent and homogeneous solution, stirring reaction 3h at room temperature, sucking filtration is precipitated, filtrate is preserved to 12h at 5 ℃, sucking filtration must precipitate again, by the precipitation vacuum drying 14h obtaining for twice, obtain white solid, with absolute ethanol washing white solid 3 times, each dehydrated alcohol 50mL that uses, obtain 4.5g6-tolysulfonyl-beta-schardinger dextrin-,
Two, take 2g6-tolysulfonyl-beta-schardinger dextrin-, 30mgKI and 0.6mL diethylenetriamine are dissolved in the N-Methyl pyrrolidone that 5mL is dry, stirring reaction 7h under 75 ℃ of conditions, obtain yellow solution, be cooled to room temperature, yellow solution is poured in 100mL dehydrated alcohol and separated out white precipitate, sucking filtration is collected white precipitate, respectively with 100mL dehydrated alcohol and the washing of 50mL ether, vacuum drying obtains 1.7g product A, product A is dissolved with 10mL distilled water, with hydrogen ion exchange resin post, carry out eluting, eluant is followed successively by 400mL distilled water, the ammonia of 400mL1mol/L, the eluent of collecting ammonia part is concentrated, vacuum drying, obtain product B, product B is dissolved in to 20mL distilled water, decompression, concentrate drying obtains 1.5g6-diethylenetriamine chain-beta-schardinger dextrin-,
Three, take 2.6g6-tolysulfonyl-beta-schardinger dextrin-and 2.4g6-diethylenetriamine chain-beta-schardinger dextrin-, be dissolved in 80mL DMF, at N
2under protection and 80 ℃ of condition of water bath heating, reaction is stirred 3 days, reaction solution is concentrated into 20mL, then join in 300mL acetone and produce and precipitate, sucking filtration also precipitates 5 times with acetone cyclic washing, precipitation is dissolved in 15mL distilled water and with water cellulose filter membrane again and filters, filtrate is dried to obtain 3.8g diethylenetriamine bridging-beta-schardinger dextrin-dimer;
Four, produce clathrate: take diethylenetriamine bridging-beta-schardinger dextrin-dimer of 0.7g in sample cell, add 5mL distilled water to make its dissolving, add again 40mg belulinic acid Betulinic acid, after sonic oscillation 40min, on agitator, continue vibration 7 days, be placed on afterwards in high speed centrifuge with the centrifugal 30min of 10000rpm, getting supernatant filters with water cellulose filter membrane, again filtrate is dried, obtains white powder, be hydrophilic belulinic acid Betulinic acid preparation.
The dry freeze-drying that adopts described in step 4, is specially: filtrate is placed in freezer dryer, and at-50 ℃ of precooling 6h, vacuum drying 24h under 200Pa.
Utilize infrared, mass spectrum, thermogravimetric analysis, X-ray diffraction and nuclear magnetic resonance, NMR to characterize, and the solution after utilizing super effect liquid phase to the de-bag of clathrate carry out the detection of belulinic acid Betulinic acid content.Through super effect liquid phase, recording belulinic acid Betulinic acid content in hydrophilic belulinic acid Betulinic acid preparation prepared by the present embodiment is 336 μ g/mL.
Fig. 7 is the infrared spectrum of the hydrophilic belulinic acid Betulinic acid preparation prepared of belulinic acid Betulinic acid, diethylenetriamine bridging beta-schardinger dextrin-dimer, the two physical mixture and the present embodiment.In Fig. 7, a is belulinic acid Betulinic acid, and b is diethylenetriamine bridging beta-schardinger dextrin-dimer, and c is the two physical mixture, and d is hydrophilic belulinic acid Betulinic acid preparation.The spectrogram that can find out physical mixture is the stack of belulinic acid Betulinic acid and diethylenetriamine bridging beta-schardinger dextrin-dimer absworption peak, has retained the C=O stretching vibration absworption peak (1685.65cm of belulinic acid Betulinic acid association carboxylic acid
-1) and C-O stretching vibration peak (1236.35cm
-1), and hydrophilic belulinic acid Betulinic acid preparation spectrogram is obviously different, there is no these two characteristic absorption peaks of belulinic acid Betulinic acid, shows that belulinic acid Betulinic acid is by enclose, has formed clathrate.
Fig. 8 is the X-ray diffractogram of the hydrophilic belulinic acid Betulinic acid preparation prepared of belulinic acid Betulinic acid, diethylenetriamine bridging beta-schardinger dextrin-dimer, the two physical mixture and the present embodiment.In Fig. 8, a is belulinic acid Betulinic acid, and b is diethylenetriamine bridging beta-schardinger dextrin-dimer, and c is the two physical mixture, and d is hydrophilic belulinic acid Betulinic acid preparation.As can be seen from the figure belulinic acid Betulinic acid and diethylenetriamine bridging beta-schardinger dextrin-dimer have visibly different diffraction maximum separately, physical mixture spectrogram is the stack of two components, the characteristic diffraction peak that has retained belulinic acid Betulinic acid in physical mixture, and the characteristic diffraction peak of belulinic acid Betulinic acid disappears in hydrophilic belulinic acid Betulinic acid preparation, illustrate and formed new thing phase, i.e. clathrate.
Fig. 9 is the mass spectrum of the de-thing of contracting for fixed output quotas of the hydrophilic belulinic acid Betulinic acid preparation prepared of the present embodiment, can see the molecular ion peak that in belulinic acid Betulinic acid standard substance figure, stable existence mass-to-charge ratio is 454.8, in the de-thing of contracting for fixed output quotas of hydrophilic belulinic acid Betulinic acid preparation, exist 455.2 molecular ion peak and intensity large, illustrate to take off in the thing of contracting for fixed output quotas and contain belulinic acid Betulinic acid.
Figure 10 is the UPLC figure of the de-thing of contracting for fixed output quotas of the hydrophilic belulinic acid Betulinic acid preparation prepared of the present embodiment, de-belulinic acid Betulinic acid integration peak area of contracting for fixed output quotas in thing is 76094, be brought in belulinic acid Betulinic acid standard curve linear equation A=6705.25C (μ g/mL)-14037.09 (correlation coefficient 0.9997), obtaining concentration is 13.44ppm, and known belulinic acid Betulinic acid dissolubility in water increases to 336.00 μ g/mL as calculated.
Claims (7)
1. a preparation method for hydrophilic belulinic acid Betulinic acid preparation, is characterized in that the method carries out according to the following steps:
One, taking 40～60g beta-schardinger dextrin-joins in 400～500mL distilled water, obtain cyclodextrin solution, getting 6～7g NaOH is dissolved in 20～25mL distilled water, obtain NaOH solution, the NaOH solution of preparation is dropwise joined in cyclodextrin solution, obtain transparent and homogeneous solution, getting 7～10g p-methyl benzene sulfonic chloride is dissolved in 30～40mL acetonitrile, obtain mixed solution, mixed solution is dropwise joined in transparent and homogeneous solution, stirring reaction 2～4h at room temperature, sucking filtration is precipitated, filtrate is preserved to 12h at 0～5 ℃, sucking filtration must precipitate again, by the precipitation vacuum drying 12～16h obtaining for twice, obtain white solid, with absolute ethanol washing white solid 3～5 times, each dehydrated alcohol 50～80mL that uses, obtain 6-tolysulfonyl-beta-schardinger dextrin-,
Two, take 1.5～3g6-tolysulfonyl-beta-schardinger dextrin-, 20～30mgKI and 0.5～1.0mL diethylenetriamine are dissolved in the N-Methyl pyrrolidone that 5～8mL is dry, stirring reaction 4～7h under 70～80 ℃ of conditions, obtain yellow solution, be cooled to room temperature, yellow solution is poured in 100～150mL dehydrated alcohol and separated out white precipitate, sucking filtration is collected white precipitate, respectively with 100～150mL dehydrated alcohol and the washing of 50～80mL ether, vacuum drying obtains 1.3～2.5g product A, product A is dissolved with 8～12mL distilled water, with hydrogen ion exchange resin post, carry out eluting, eluant is followed successively by 400～500mL distilled water, the ammonia of 400～500mL1mol/L, the eluent of collecting ammonia part is concentrated, vacuum drying, obtain product B, product B is dissolved in to 20～30mL distilled water, decompression, concentrate drying obtains 6-diethylenetriamine chain-beta-schardinger dextrin-,
Three, take 2.6～3.9g6-tolysulfonyl-beta-schardinger dextrin-and 1.2～2.4g6-diethylenetriamine chain-beta-schardinger dextrin-, be dissolved in 80～100mLN, in dinethylformamide, at N
2under protection and 75～85 ℃ of condition of water bath heating, reaction is stirred 2～3 days, reaction solution is concentrated into 20～30mL, then join in 300～400mL acetone and produce and precipitate, sucking filtration also precipitates 3～5 times with acetone cyclic washing, precipitation is dissolved in 10～15mL distilled water and with water cellulose filter membrane again and filters, filtrate is dried to obtain diethylenetriamine bridging-beta-schardinger dextrin-dimer;
Four, produce clathrate: take 6-diethylenetriamine chain-beta-schardinger dextrin-of 0.5～0.7g or 0.7～1.0g diethylenetriamine bridging-beta-schardinger dextrin-dimer in sample cell, add 5～6mL distilled water to make its dissolving, add again 30～40mg belulinic acid Betulinic acid, after sonic oscillation 20～60min, on agitator, continue vibration 6～7 days, be placed on afterwards in high speed centrifuge with the centrifugal 15～30min of 10000～12000rpm, getting supernatant filters with water cellulose filter membrane, again filtrate is dried, obtain white powder, be hydrophilic belulinic acid Betulinic acid preparation.
2. the preparation method of a kind of hydrophilic belulinic acid Betulinic acid preparation according to claim 1, is characterized in that in step 2, taking 2g6-tolysulfonyl-beta-schardinger dextrin-, 25mgKI and 0.57mL diethylenetriamine is dissolved in the N-Methyl pyrrolidone that 5mL is dry.
3. the preparation method of a kind of hydrophilic belulinic acid Betulinic acid preparation according to claim 1 and 2, is characterized in that described in step 2 that resin column specification is: 25 * 450mm, and 724 weak-acid cation-exchange resins, resin filling height is 250mm.
4. the preparation method of a kind of hydrophilic belulinic acid Betulinic acid preparation according to claim 3, is characterized in that taking 2.6g6-tolysulfonyl-beta-schardinger dextrin-and 2.4g6-diethylenetriamine chain-beta-schardinger dextrin-in step 3, is dissolved in 80mLN, in dinethylformamide.
5. the preparation method of a kind of hydrophilic belulinic acid Betulinic acid preparation according to claim 4, is characterized in that taking in step 4 6-diethylenetriamine chain-beta-schardinger dextrin-of 0.6g or 0.8g diethylenetriamine bridging-beta-schardinger dextrin-dimer in sample cell.
6. the preparation method of a kind of hydrophilic belulinic acid Betulinic acid preparation according to claim 5, is characterized in that adding in step 4 35mg belulinic acid Betulinic acid.
7. the preparation method of a kind of hydrophilic belulinic acid Betulinic acid preparation according to claim 6, it is characterized in that being dried employing freeze-drying described in step 4, be specially: filtrate is placed in freezer dryer, at-50 ℃ of precooling 6～8h, vacuum drying 24～36h under 30～200Pa.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104826123A (en) * | 2015-04-10 | 2015-08-12 | 昆明理工大学 | Clathrate compound of oleanolic acid and amine cyclodextrin |
| CN114225050A (en) * | 2022-01-29 | 2022-03-25 | 国家卫生健康委科学技术研究所 | Preparation and application of ethinylestradiol cyclodextrin compound |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102716491A (en) * | 2012-07-02 | 2012-10-10 | 昆明理工大学 | Clathrate compound of artemisinin series and alkaline cyclodextrin and method for preparing same |
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Non-Patent Citations (2)
| Title |
|---|
| YAN-FANG SUN ET AL.: "Apoptosis of human breast cancer cells induced by microencapsulated betulinic acid from sour jujube fruits through the mitochondria transduction pathway", 《FOOD CHEMISTRY》 * |
| 申刚义 等: "新型超分子化合物桥联β-环糊精合成研究进展", 《化学通报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104826123A (en) * | 2015-04-10 | 2015-08-12 | 昆明理工大学 | Clathrate compound of oleanolic acid and amine cyclodextrin |
| CN104826123B (en) * | 2015-04-10 | 2017-11-10 | 昆明理工大学 | A kind of inclusion compound of oleanolic acid and amine cyclodextrin |
| CN114225050A (en) * | 2022-01-29 | 2022-03-25 | 国家卫生健康委科学技术研究所 | Preparation and application of ethinylestradiol cyclodextrin compound |
| WO2023143306A1 (en) * | 2022-01-29 | 2023-08-03 | 国家卫生健康委科学技术研究所 | Preparation for ethinylestradiol-cyclodextrin complex and use thereof |
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