CN105968120B - Preparation method of tetrandrine and tetrandrine B - Google Patents
Preparation method of tetrandrine and tetrandrine B Download PDFInfo
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- CN105968120B CN105968120B CN201610366510.XA CN201610366510A CN105968120B CN 105968120 B CN105968120 B CN 105968120B CN 201610366510 A CN201610366510 A CN 201610366510A CN 105968120 B CN105968120 B CN 105968120B
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
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Abstract
The invention discloses a preparation method of tetrandrine and tetrandrine, which belongs to the technical field of medicine preparation, and is prepared by adopting lipophilic tertiary amine total alkali through 2 steps, 1) separating by using a medium-pressure silica gel chromatographic column method, and eluting by using petroleum ether-ethyl acetate-diethylamine; 2) and purifying by combining with preparative high performance liquid chromatography, taking methanol-water as a mobile phase, and finally respectively concentrating and evaporating eluent to dryness.
Description
Technical Field
The invention relates to a preparation method of a medicament, in particular to a preparation method of tetrandrine and tetrandrine.
Background
Radix Stephaniae Tetrandrae, rhizoma Stephaniae Tetrandrae, Bufo siccus, and herba Ardisiae Japonicae are dry root tuber of radix Stephaniae Tetrandrae of Stephania of Menispermaceae, and contain polysaccharide, alkaloid, etc., and its main effective components are tetrandrine and tetrandrine. Tetrandrine has antiinflammatory, analgesic, blood pressure lowering, antitumor, anti-hypoxic pulmonary hypertension, and pulmonary fibrosis inhibiting effects. Tetrandrine is a natural calcium antagonist, and has therapeutic effect on hypertension, diabetes, hepatic fibrosis, etc., and also has brain cell protecting effect and broad-spectrum antiinflammatory and antibacterial effects. The existing method for preparing tetrandrine and tetrandrine has the common defects of complex process, high production cost and low preparation efficiency.
Disclosure of Invention
The invention aims to overcome the defects of complex process and high preparation cost of the existing method and provide a novel preparation method of tetrandrine and tetrandrine.
In order to achieve the purpose, the technical scheme is as follows:
a method for preparing tetrandrine and tetrandrine B comprises the following steps:
1) separating tetrandrine and tetrandrine by medium pressure silica gel chromatography, loading a column with volume of 49mm × 460mm, loading lipophilic tertiary amine total alkali into the column, eluting with petroleum ether-ethyl acetate-diethylamine at flow rate of 80 mL/min, collecting eluate, inspecting by TLC, mixing the same components, recovering solvent to obtain two components Fr 1 and Fr 2;
2) preparing high performance liquid chromatography for purifying tetrandrine and tetrandrine: and (2) aiming at the components Fr 1 and Fr 2, adopting a YMC-pack chromatographic column 250 x 10mmD, S-5 mu m and 12 nm, selecting 230 nm as a detection wavelength, the flow rate of 4 mL/min and methanol-water as a mobile phase, carrying out sample injection separation, collecting the separated eluates of two main peaks, and respectively concentrating and evaporating the eluates to dryness to obtain the compound.
Further, in the step 1), the silica gel used by the chromatographic column is 300-400 mesh.
Further, in the step 1), the ratio of petroleum ether-ethyl acetate-diethylamine is 9:1:0.1-5:1: 0.1.
Further, in the step 1), the solvent is recovered by using a rotary evaporator.
Further, in the step 2), the components Fr 1 and Fr 2 are filtered by a 0.22 μm microporous filter membrane before being injected.
Further, in the step 2), the ratio of methanol to water is 17: 3.
Further, in the step 2), the sample injection amount is 0.5 mL.
The beneficial effect who adopts above-mentioned scheme does: the tetrandrine and the tetrandrine B prepared by combining the medium-pressure silica gel chromatographic column method with the high performance liquid chromatography have the advantages of convenience and rapidness in operation and high purity, and two products can be obtained by one preparation method, so that the extraction rate is high.
Drawings
FIG. 1 is the TLC bismuth potassium iodide and TLC 254nm purple outer color chart of tetrandrine and tetrandrine B prepared by the embodiment of the invention.
FIG. 2 shows tetrandrine prepared according to the present invention1H-NMR chart.
FIG. 3 shows the preparation of tetrandrine1H-NMR chart.
Detailed Description
The present invention is further described with reference to the following embodiments and drawings, but the present invention is not limited to the following embodiments, and it is anticipated that one skilled in the art may make various changes in the embodiments in combination with the prior art.
A method for preparing tetrandrine and tetrandrine B comprises the following steps:
1) the method for separating tetrandrine and tetrandrine B by medium pressure silica gel chromatographic column comprises loading a chromatographic column with volume of 49mm × 460mm, wherein silica gel used in the chromatographic column is 300-400 mesh, loading lipophilic tertiary amine total alkali into the column, eluting with petroleum ether-ethyl acetate-diethylamine (9:1:0.1-5:1: 0.1) at flow rate of 80 mL/min, detecting the collected eluate by TLC, mixing the components, recovering solvent with rotary evaporator to obtain two main components Fr 1 and Fr 2 with high content of tetrandrine and tetrandrine B respectively.
2) And preparing high performance liquid chromatography for purifying tetrandrine and tetrandrine: adopting YMC-pack chromatographic column (250 x 10mmD, S-5 μm,12 nm), selecting 230 nm as detection wavelength, flow rate of 4 mL/min, methanol (A) -water (B) as mobile phase, and continuously adjusting methanol (A) -water (B) ratio and sample amount (sample passes through 0.22 μm microporous filter membrane before sample injection). Until a suitable chromatographic condition is found, the ratio of the methanol to the water is 85 percent to 15 percent, the sample amount is 0.5 mL, the peak pattern is better, and the separation degree is high. The eluate of the two main peaks obtained by separation was collected and the remaining components were discarded. And (3) concentrating and evaporating the collected eluent respectively, dissolving a proper amount of product with methanol, comparing TLC (thin layer chromatography) with a standard substance, and performing chromatography with petroleum ether: ethyl acetate: diethylamine (3: 2: 1) as developing agent, and bismuth potassium iodide as color developing agent. Inspection under 254nm UV.
The tetrandrine and tetrandrine B prepared by the invention have the same spot color and position with the reference substance, and have single component. As shown in FIG. 1, TLC bismuth potassium iodide chromogenic diagram is arranged on the left side, Han TLC 254nm purple outer chromogenic diagram is arranged on the right side, and tetrandrine, tetrandrine standard substance, tetrandrine and tetrandrine standard substance are arranged on the spots from left to right in sequence.
As shown in figure 2, the prepared tetrandrine1H-NMR results: a white-like powder.1H-NMR(400 MHz, CDCl3):2.33(s,3H), 2.41 ～ 2.47(m, 1H),2.52(d,J=12.0 Hz, 1H), 2.62( s, 3H), 3.19(s,3H), 3.25(dd,J=12.0,8.0 Hz, 1H), 3.37(s, 3H), 3.43～3.48(m, 1H), 3.51～3.56(m, 1H), 3.75(s, 3H), 3.88(dd,J=12.0,4.0Hz, 1H), 3.93(s,3H), 5.99(s, 1H),6.29(s, 1H), 6.30(m, 2H), 6.51(s,1H), 6.55(s, 1H), 6.81(d,J=8.0 Hz, 1H),6.86(dd,J=8.0, 4.0 Hz, 1H), 6.87(s, 1H), 6.89(s, 1H), 7.13(dd,J=8.0, 4.0Hz, 1H), 7.34(d,J=8.0 Hz, 1H) 。
As shown in figure 3, the obtained tetrandrine is white powder.1H-NMR(400 MHz, CDCl3): 2.33(s,3H),2.41 ～ 2.45(m, 2H),2.56(d,J=16.0 Hz, 1H), 2.63( s, 3H), 3.26(dd,J=12.0,8.0 Hz, 1H), 3.35(s, 3H), 3.48～3.54(m, 2H), 3.77(s, 3H), 3.88-3.91(m, 1H),3.92(s,3H), 6.05(s, 1H), 6.29(s, 1H), 6.32(m, 1H), 6.52(s,1H), 6.56(s, 1H),6.81(d,J=8.0 Hz, 1H), 6.86(o, 1H), 6.87(o, 1H), 6.89(o, 1H), 7.13(dd,J=8.0,4.0 Hz, 1H), 7.34(d,J=8.0 Hz, 1H) 。
The tetrandrine and the tetrandrine are different from each other in the substituent group at the 7-position, wherein the tetrandrine is methoxy, and the tetrandrine is phenolic hydroxyl. In that1In H-NMR, 4 methoxy signals, 3.19(s, 3H, H-7), 3.37(s, 3H, H-6 ´)3.75(s, 3H, H-6), 3.93(s,3H, H-12) were present between 3.0 and 4.0, whereas in hanfangchin, only 3 methoxy signals, 3.35(s,3H), 3.77(s, 3H), 3.92(s, 3H), were present at positions 6, 6 and 12, respectively, so that the signal was derived from 6 ´, 6 and 12, respectively1The element A and the element B can be easily distinguished by H-NMR.
Claims (1)
1. A preparation method of tetrandrine and tetrandrine B is characterized by comprising the following steps:
1) loading a chromatographic column with the volume of 49mm × 460mm, loading lipophilic tertiary amine total alkali into the chromatographic column, eluting by petroleum ether-ethyl acetate-diethylamine with the flow rate of 80 mL/min, collecting eluent, inspecting by TLC, merging the same components, recovering the solvent by using a rotary evaporation instrument to obtain two components Fr 1 and Fr 2, wherein the silica gel used by the chromatographic column is 300-mesh and 400-mesh;
2) preparing high performance liquid chromatography for purifying tetrandrine and tetrandrine: adopting YMC-pack chromatographic columns 250 x 10mmD, S-5 mu m and 12 nm aiming at the components Fr 1 and Fr 2, selecting 230 nm as a detection wavelength, the flow rate of 4 mL/min and methanol-water as a mobile phase, carrying out sample injection separation, collecting the eluates of two main peaks obtained by separation, and respectively concentrating and evaporating the eluates to dryness to obtain the compound preparation; the ratio of the methanol to the water is 17: 3; filtering the components Fr 1 and Fr 2 with a 0.22 μm microporous filter membrane before sample injection; the sample injection amount is 0.5 mL.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1850824A (en) * | 2006-04-27 | 2006-10-25 | 富力 | Benzene-free tetrandrine and its preparing method |
| CN101012229A (en) * | 2007-01-26 | 2007-08-08 | 北海阳光药业有限公司 | Hanfangchin A without benzene and chloroform and preparing method thereof |
| CN101033229A (en) * | 2007-02-09 | 2007-09-12 | 南开大学 | Separation of monosomic tetrandrine from tetrandrine total alkaloid by adsorption resin method |
| CN101288695A (en) * | 2007-04-16 | 2008-10-22 | 中国科学院成都生物研究所 | Preparation technique of alkaloids from Stephania tetrandra |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1850824A (en) * | 2006-04-27 | 2006-10-25 | 富力 | Benzene-free tetrandrine and its preparing method |
| CN101012229A (en) * | 2007-01-26 | 2007-08-08 | 北海阳光药业有限公司 | Hanfangchin A without benzene and chloroform and preparing method thereof |
| CN101033229A (en) * | 2007-02-09 | 2007-09-12 | 南开大学 | Separation of monosomic tetrandrine from tetrandrine total alkaloid by adsorption resin method |
| CN101288695A (en) * | 2007-04-16 | 2008-10-22 | 中国科学院成都生物研究所 | Preparation technique of alkaloids from Stephania tetrandra |
Non-Patent Citations (1)
| Title |
|---|
| TLC法分离汉防己中汉防己甲素、乙素;徐应淑 等;《遵义医学院学报》;20090630;第32卷(第3期);第296-297页，第2.2.2节和3.3节 * |
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