CN108623524B - Imidazole dimer alkaloid, and preparation method and application thereof - Google Patents
Imidazole dimer alkaloid, and preparation method and application thereof Download PDFInfo
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- 229930013930 alkaloid Natural products 0.000 title claims abstract description 49
- -1 Imidazole dimer alkaloid Chemical class 0.000 title claims abstract description 34
- 238000002360 preparation method Methods 0.000 title claims description 11
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- 230000000324 neuroprotective effect Effects 0.000 claims abstract description 5
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- 238000010828 elution Methods 0.000 claims description 36
- 238000010898 silica gel chromatography Methods 0.000 claims description 34
- WGLUMOCWFMKWIL-UHFFFAOYSA-N dichloromethane;methanol Chemical compound OC.ClCCl WGLUMOCWFMKWIL-UHFFFAOYSA-N 0.000 claims description 30
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 27
- 244000269722 Thea sinensis Species 0.000 claims description 25
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- 238000004440 column chromatography Methods 0.000 claims description 16
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- 150000002460 imidazoles Chemical class 0.000 description 13
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- 230000000975 bioactive effect Effects 0.000 description 3
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- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 2
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- YAPQBXQYLJRXSA-UHFFFAOYSA-N theobromine Chemical compound CN1C(=O)NC(=O)C2=C1N=CN2C YAPQBXQYLJRXSA-UHFFFAOYSA-N 0.000 description 2
- ZFXYFBGIUFBOJW-UHFFFAOYSA-N theophylline Chemical compound O=C1N(C)C(=O)N(C)C2=C1NC=N2 ZFXYFBGIUFBOJW-UHFFFAOYSA-N 0.000 description 2
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- ZRJBHWIHUMBLCN-YQEJDHNASA-N huperzine A Chemical group N1C(=O)C=CC2=C1C[C@H]1\C(=C/C)[C@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-YQEJDHNASA-N 0.000 description 1
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- 230000001681 protective effect Effects 0.000 description 1
- 229930002161 purine alkaloid Natural products 0.000 description 1
- 150000003212 purines Chemical class 0.000 description 1
- 150000003230 pyrimidines Chemical class 0.000 description 1
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Images
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
- C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
- C07D233/66—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D233/88—Nitrogen atoms, e.g. allantoin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
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- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
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Abstract
The invention discloses an imidazole dimer alkaloid capable of being used for preparing a neuroprotective drug, which is represented by a structure shown in a formula I, a formula II or a formula III:
Description
Technical Field
The invention relates to the field of natural products, in particular to imidazole dimer alkaloid and a preparation method and application thereof.
Background
The tea contains rich alkaloid, which accounts for 3% -5% of the dry matter of the tea. Most of them are purine alkaloids, and a small amount of pyrimidine alkaloids. Wherein the caffeine accounts for 3-4%, the theobromine accounts for 0.15-0.2%, and the theophylline accounts for 0.02-0.04%. Research shows that caffeine is an important flavor substance in tea. In addition, caffeine also has the pharmacological functions of tonifying heart, promoting urination, removing toxic substances, relieving asthma, etc.
The black tea is fully fermented, and the processing of the keemun black tea mainly comprises the working procedures of withering, rolling, fermenting, drying and the like, and finally the color of the dry tea and the color of the brewed tea soup are mainly red. Researches prove that the black tea has good free radical scavenging activity, and can promote appetite and help gastrointestinal digestion. Recent pharmacological research shows that the Chinese medicinal composition has various pharmacological functions of cancer prevention, cancer resistance, antibiosis, ultraviolet irradiation resistance, oxidation resistance, atherosclerosis resistance, caries resistance, tooth protection and the like.
The reports on the bioactive alkaloid chemical components of Keemun black tea are relatively few, and if the bioactive alkaloid derivative components can be successfully researched and developed from Keemun black tea, the bioactive alkaloid derivative components can make important contributions to the fields of agriculture, medicines and the like.
Disclosure of Invention
The technical problem to be solved by the invention is to provide an imidazole dimer alkaloid capable of protecting nerves.
In order to solve the technical problems, the invention adopts the following technical scheme: an imidazole dimer alkaloid represented by the structure of formula I or formula II or formula III:
the invention also provides application of the imidazole dimer alkaloid in preparation of neuroprotective drugs.
In particular to a medicine with neuroprotective effect, which is prepared from the imidazole dimer alkaloid and pharmaceutic adjuvant.
The pharmaceutical dosage forms of the medicament include oral and injectable forms. The oral dosage forms comprise tablets, capsules, granules, dripping pills and the like; the injection type comprises injection, racemic mixture and the like. The preparation method refers to the conventional method in the pharmaceutical field, and the used pharmaceutic adjuvant selects the general adjuvant in the pharmaceutical field according to different dosage forms.
The invention also provides a preparation method of the imidazole dimer alkaloid, which comprises the following steps:
(1) raw material crushing
Pulverizing Keemun black tea to obtain Keemun black tea powder;
(2) leaching
Extracting the Keemun black tea powder with ethanol water solution to obtain extractive solution, and drying the extractive solution to obtain Keemun black tea extract;
(3) separating and purifying
Dissolving Keemun black tea extract with methanol water solution, concentrating to remove most of methanol, extracting with petroleum ether to remove small polar substances and pigment, and extracting with dichloromethane to obtain dichloromethane part extract A;
dissolving the dichloromethane part extract A with water, adding tartaric acid aqueous solution, removing insoluble substances, extracting with dichloromethane, removing water layer, adding equal volume of sodium hydroxide aqueous solution, extracting to obtain dichloromethane layer and alkaline water layer, adding ammonium chloride into the alkaline water layer, and extracting with dichloromethane to obtain dichloromethane part extract B;
and (3) separating and purifying the dichloromethane part extract B to obtain the imidazole dimer alkaloid.
Further, in the step (2), the concentration of the ethanol aqueous solution used is 95%.
Further, in the step (3), the concentration of the tartaric acid aqueous solution used is 2%, and the concentration of the sodium hydroxide aqueous solution is 1-2%.
Further, in the step (3), the separation and purification treatment comprises silica gel column chromatography, Sephadex LH-20 gel column chromatography and Toyopearl column chromatography.
Further, the specific steps of the separation and purification treatment are as follows: dissolving the dichloromethane part extract B, then carrying out first silica gel column chromatography, carrying out first silica gel column chromatography by taking dichloromethane-methanol volume ratio of 500:1 to 1:1 as gradient elution, collecting dichloromethane-methanol volume ratio of 20:1 to 10:1 as elution component, carrying out second silica gel column chromatography by taking dichloromethane-methanol volume ratio of 50:1 to 1:1 as gradient elution, collecting dichloromethane-methanol volume ratio of 10:1 as elution component, carrying out third silica gel column chromatography by taking dichloromethane-methanol volume ratio of 10:1 to 1:1 as gradient elution component, collecting elution component, carrying out Toyopearl column chromatography purification, and obtaining the imidazole dimer alkaloid shown in formula I;
performing fourth silica gel column chromatography on a dichloromethane-methanol volume ratio of 30:1 to 20:1 elution component obtained by the first silica gel column chromatography, performing gradient elution on the fourth silica gel column chromatography by using a dichloromethane-methanol volume ratio of 70:1 to 1:1, collecting an elution component with a dichloromethane-methanol volume ratio of 10:1, performing Sephadex LH-20 gel column chromatography by using methanol to flush the column, collecting a first eluted component, and performing Toyopearl column chromatography purification to obtain the imidazole dimer alkaloid shown in the formula II;
and (3) carrying out fifth silica gel column chromatography on the dichloromethane-methanol volume ratio 20:1 elution component obtained by the fourth silica gel column chromatography, carrying out gradient elution on the fifth silica gel column chromatography by using the dichloromethane-methanol volume ratio of 40:1 to 1:1, collecting the elution component, and carrying out Toyopearl column chromatography purification to obtain the imidazole dimer alkaloid shown in the formula III.
The term "first.
The invention has the beneficial effects that:
the imidazole dimer alkaloid with medical activity provided by the invention has a certain protection effect in a nerve cell damage model induced by hydrogen peroxide, can be used for preparing neuroprotective drugs, and has important significance in the fields of agriculture and medicine; and provides wider prospect for effectively developing and utilizing Keemun black tea.
The preparation method of the imidazole dimer alkaloid has the advantages of simple process, easy implementation, low cost and good application prospect.
Drawings
FIG. 1 is a diagram of the x-ray crystal structure of imidazole dimer alkaloids of formula I.
FIG. 2 is a diagram of the x-ray crystal structure of imidazole dimer alkaloid of formula II.
FIG. 3 is a schematic diagram of the experiment of the inhibition of hydrogen peroxide induced human nerve cell damage by imidazole dimer alkaloids shown in formula I, formula II and formula III.
Detailed Description
The invention is further described below with reference to the following examples:
this section generally describes the materials used in the experiments of the present invention, as well as the experimental methods. Although many materials and methods of operation are known in the art for the purpose of carrying out the invention, the invention is described herein in detail as much as possible. It will be apparent to those skilled in the art that the materials, equipment, and methods of operation used in the present invention are well known in the art to which the invention pertains, unless otherwise specified.
Example 1
Preparation of imidazole dimer alkaloids
1.1 description of imidazole dimer alkaloids
The imidazole dimer alkaloid has a structure shown in formula I, formula II or formula III:
1.2 preparation method and results
(1) Raw material crushing
Pulverizing 200 kg Keemun black tea into powder, and sieving with 10.5mm mesh sieve to obtain Keemun black tea powder;
(2) leaching
Adding Keemun black tea powder into 1000 kg of 95% ethanol aqueous solution, stirring and extracting for 10 hours, filtering to obtain an extracting solution, concentrating the extracting solution to 400L, passing through a 200-mesh screen, continuously concentrating, passing through a 200-mesh screen, finally performing spray drying on the concentrated solution, collecting dried powder, and passing through a 40-mesh screen to obtain a black tea extract, wherein the air inlet temperature is 190 ℃, the air outlet temperature is 90 ℃;
(3) separating and purifying
Dissolving Keemun black tea extract with 70% methanol water solution, concentrating to remove most of methanol (as much as possible, so as not to affect subsequent extraction layering), extracting with petroleum ether to remove small polar substances and pigment, and extracting with dichloromethane to obtain dichloromethane part extract A;
dissolving the dichloromethane part extract A with water, adding 2% tartaric acid aqueous solution, removing insoluble substances, then extracting with dichloromethane, removing the water layer, adding 1% -2% sodium hydroxide aqueous solution with the same volume to extract to obtain a dichloromethane layer and an alkaline water layer, adding ammonium chloride into the alkaline water layer, and then extracting with dichloromethane to obtain a dichloromethane part extract B;
dissolving 500 g of dichloromethane part extract B by using chloroform, mixing the mixture with a sample, performing silica gel column chromatography for the first time, performing gradient elution by using a dichloromethane-methanol volume ratio of 500:1 to 1:1 on the silica gel column chromatography for the first time, collecting an elution component with a dichloromethane-methanol volume ratio of 20:1 to 10:1, performing silica gel column chromatography for the second time, performing gradient elution by using a dichloromethane-methanol volume ratio of 50:1 to 1:1 on the silica gel column chromatography for the second time, collecting an elution component with a dichloromethane-methanol volume ratio of 10:1 to 1:1 on the silica gel column chromatography for the third time, performing gradient elution by using a dichloromethane-methanol volume ratio of 10:1 to 1:1 on the silica gel column chromatography for the third time, collecting an elution component (the column chromatography mainly removes a small amount of impurities, has no components, mainly only one component, but has insufficient purity, and the subsequent Toyopeal purification is used for continuously purifying a compound with insufficient purity), obtaining 12.0mg of imidazole dimer alkaloid shown in the formula I;
performing fourth silica gel column chromatography on a dichloromethane-methanol volume ratio of 30:1 to 20:1 elution component obtained by the first silica gel column chromatography, performing gradient elution on the fourth silica gel column chromatography by using a dichloromethane-methanol volume ratio of 70:1 to 1:1, collecting an elution component with a dichloromethane-methanol volume ratio of 10:1, performing Sephadex LH-20 gel column chromatography by using methanol to flush a column, collecting a first eluted component, and performing Toyopearl column chromatography purification to obtain 11.5mg of imidazole dimer alkaloid shown in formula II;
and (3) carrying out fifth silica gel column chromatography on the dichloromethane-methanol volume ratio 20:1 elution component obtained by the fourth silica gel column chromatography, carrying out gradient elution on the fifth silica gel column chromatography by using the dichloromethane-methanol volume ratio 40:1 to 1:1, collecting the elution component (the same as the non-component here) and carrying out Toyopearl column chromatography purification to obtain 8.0mg of the imidazole dimer alkaloid shown in the formula III.
1.3 Property verification of imidazole dimer alkaloid
1.3.1 characteristics of imidazole dimer alkaloids of formula I are as follows:
1) white powder, soluble in methanol and water, insoluble in DMSO;
2) melting point 257-258 ℃; HR-ESI-MS: M/z 349.20895([ M + H)]+,calcd for C15H25N8O2 +,349.21005),371.19019([M+Na]+,calcd for C15H24N8O2Na+,371.19199). The nmr spectral data are shown in table 1; crystal data and structure refinement are shown in table 2; the X-ray crystal structure is shown in figure 1.
TABLE 1 Nuclear magnetic resonance spectroscopy data for imidazole dimer alkaloids of formula I (1H NMR at 600MHz, 13C NMR at 125MHz, delta units in ppm, coupling constant J units in Hz, solvent deuterated methanol).
| Position of | δH(J，Hz) | |
| 2 | 7.54s | 137.4 |
| 2' | 7.54s | 137.4 |
| 4 | 138.6 | |
| 4' | 138.6 | |
| 5 | 124.1 | |
| 5' | 124.1 | |
| 6 | 3.56s | 32.9 |
| 6' | 3.56s | 32.9 |
| 8 | 2.95s | 36.6 |
| 8' | 2.95s | 36.6 |
| 9 | 160.8 | |
| 9' | 160.8 | |
| 10 | 5.98br s | |
| 10' | 5.98br s | |
| 11 | 2.63d(4.2) | 28.0 |
| 11' | 2.63d(4.2) | 28.0 |
| 12 | 3.82s | 17.6 |
TABLE 2 Crystal data for imidazole dimer alkaloids of formula I
All spectral data are passed1H-1The two-dimensional nuclear magnetic resonance spectrum attribution of H COSY, HMQC, HMBC and the like proves the structure of the obtained compound.
1.3.2 characteristics of imidazole dimer alkaloids of formula II are as follows:
1) white powder, soluble in methanol, chloroform and DMSO;
2) melting point 244-245 ℃; HR-ESI-MS: M/z 392.25102([ M + H)]+,calcd for C17H30N9O2 +,392.25225),414.23422([M+Na]+,calcd for C17H29N9O2Na+,414.23419),805.47712([2M+Na]+,calcd for C34H58N18O4Na+,805.47861). Nmr spectral data are shown in table 3; crystal data and structure refinement are shown in table 4; the X-ray crystal structure is shown in figure 2.
TABLE 3 NMR spectra data for imidazole dimer alkaloids of formula II: (1The H NMR was at 600MHz and the molecular weight,13c NMR was measured at 125MHz with δ in ppm, coupling constant J in Hz, and solvent deuterated trichloromethane).
TABLE 4 Crystal data for imidazole dimer alkaloids of formula II
All spectral data are passed1H-1The two-dimensional nuclear magnetic resonance spectrum attribution of H COSY, HMQC, HMBC and the like and the analysis of X-ray crystal data prove the structure of the obtained compound.
1.3.3 characteristics of imidazole dimer alkaloids of formula III are as follows:
1) yellowish oil, soluble in methanol, chloroform and DMSO;
2)、HR-ESI-MS:m/z 371.19084([M+Na]+,calcd for C15H24N8O2Na+,371.19199). The nmr spectral data are shown in table 5.
TABLE 5 Nuclear magnetic Co-catalysis of imidazole dimer alkaloids of formula IIIVibration spectrum data (1The H NMR was at 600MHz and the molecular weight,13c NMR was measured at 125MHz with δ in ppm, coupling constant J in Hz, and solvent deuterated DMSO).
| Position of | δH(J，Hz) | |
| 2 | 7.49s | 136.37 |
| 4 | 139.90 | |
| 5 | 121.49 | |
| 7 | 158.08 | |
| 9 | 3.90s | 47.91 |
| 11 | 151.92 | |
| 13 | 7.56s | 138.34 |
| 15 | 115.41 | |
| 16 | 160.76 | |
| N8-H | 5.66q(4.8) | |
| N17-H | 8.04q(4.8) | |
| N1-CH3 | 3.53s | 32.05 |
| N6-CH3 | 2.81s | 36.03 |
| N8-CH3 | 2.47d(4.8) | 27.59 |
| N10-CH3 | 2.51s | 43.44 |
| N14-CH3 | 3.72s | 34.74 |
| N17-CH3 | 2.72d(4.8) | 25.57 |
All spectral data are passed1H-1The two-dimensional nuclear magnetic resonance spectrum attribution of H COSY, HMQC, HMBC and the like proves the structure of the obtained compound.
Example 2
In-vitro hydrogen peroxide-induced nerve injury protection experiment of imidazole dimer alkaloid
Cell culture: human neuroblastoma (SH-SY5Y) is taken and cultured in a carbon dioxide constant temperature incubator under certain conditions (temperature 37 ℃, carbon dioxide concentration 5% and humidity 95%) by Dulbecco's Modified Eagle (DMEM) culture medium containing 10% fetal calf serum and 1% double antibody. When the cell density reaches 80% -90%, the cell is transferred into a 96-well plate, and the number of cells per well is about 2 multiplied by 104And (4) respectively. Adding 200 mu M H respectively2O2And 1. mu.M compound was added after 24 hours of culture 20. mu.L of 3- (4, 5-dimethylthiozol-2-yl) -2,5-diphenyltetrazolium bromide (MTT,5mg/mL), cultured at 37 ℃ for 4 hours, the medium was removed and 150. mu.L of dimethyl sulfoxide was added. The absorbance values were measured at 490 nm.
As shown in FIG. 3, the addition of hydrogen peroxide successfully induced nerve cell damage relative to the control (Basal), and the damage model was successfully established. The huperzine A group is added to have obvious nerve cell protection effect; the imidazole dimer alkaloid group shown in the formula I, the formula II and the formula III is added to show a good neuroprotective effect, so that the imidazole dimer alkaloid has a certain protective effect on nerve injury induced by hydrogen peroxide.
The tests prove that the imidazole dimer alkaloids prepared by the invention have stronger protection effect on nerve injury, and can be used for preparing medicines for inhibiting nerve injury.
In specific implementation, the imidazole dimer alkaloid shown as formula I, formula II and formula III can be prepared into a neuroprotective medicament according to medically acceptable dose and pharmaceutically universal auxiliary materials. The pharmaceutical dosage form comprises oral administration type, injection type, etc. The oral dosage forms comprise tablets, capsules, granules, dripping pills and the like; the injection type comprises injection, racemic mixture and the like. The specific preparation method refers to the conventional method in the pharmaceutical field.
It should be understood that the examples and embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the present disclosure, and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this disclosure.
Claims (7)
1. An imidazole dimer alkaloid characterized by being represented by the structure of formula I or formula II or formula III:
2. use of an imidazole dimer alkaloid according to claim 1 in the preparation of a neuroprotective medicament.
3. A neuroprotective agent comprising the imidazole dimer alkaloid of claim 1 and a pharmaceutically acceptable excipient.
4. The neuroprotective agent according to claim 3 in the form of oral or injectable dosage form.
5. The method of preparing an imidazole dimer alkaloid of claim 1, comprising the steps of:
(1) raw material crushing
Pulverizing Keemun black tea to obtain Keemun black tea powder;
(2) leaching
Extracting the Keemun black tea powder with ethanol water solution to obtain extractive solution, and drying the extractive solution to obtain Keemun black tea extract;
(3) separating and purifying
Dissolving Keemun black tea extract with methanol water solution, concentrating to remove most of methanol, extracting with petroleum ether to remove small polar substances and pigment, and extracting with dichloromethane to obtain dichloromethane part extract A;
dissolving the dichloromethane part extract A with water, adding tartaric acid aqueous solution, removing insoluble substances, extracting with dichloromethane, removing water layer, adding equal volume of sodium hydroxide aqueous solution, extracting to obtain dichloromethane layer and alkaline water layer, adding ammonium chloride into the alkaline water layer, and extracting with dichloromethane to obtain dichloromethane part extract B;
separating and purifying the dichloromethane part extract B to obtain the imidazole dimer alkaloid; the separation and purification treatment comprises silica gel column chromatography, Sephadex LH-20 gel column chromatography and Toyopearl column chromatography, and comprises the following specific steps:
dissolving the dichloromethane part extract B, then carrying out first silica gel column chromatography, carrying out first silica gel column chromatography by taking dichloromethane-methanol volume ratio of 500:1 to 1:1 as gradient elution, collecting dichloromethane-methanol volume ratio of 20:1 to 10:1 as elution component, carrying out second silica gel column chromatography by taking dichloromethane-methanol volume ratio of 50:1 to 1:1 as gradient elution, collecting dichloromethane-methanol volume ratio of 10:1 as elution component, carrying out third silica gel column chromatography by taking dichloromethane-methanol volume ratio of 10:1 to 1:1 as gradient elution component, collecting elution component, carrying out Toyopearl column chromatography purification, and obtaining the imidazole dimer alkaloid shown in formula I;
performing fourth silica gel column chromatography on a dichloromethane-methanol volume ratio of 30:1 to 20:1 elution component obtained by the first silica gel column chromatography, performing gradient elution on the fourth silica gel column chromatography by using a dichloromethane-methanol volume ratio of 70:1 to 1:1, collecting an elution component with a dichloromethane-methanol volume ratio of 10:1, performing Sephadex LH-20 gel column chromatography by using methanol to flush the column, collecting a first eluted component, and performing Toyopearl column chromatography purification to obtain the imidazole dimer alkaloid shown in the formula II;
and (3) carrying out fifth silica gel column chromatography on the dichloromethane-methanol volume ratio 20:1 elution component obtained by the fourth silica gel column chromatography, carrying out gradient elution on the fifth silica gel column chromatography by using the dichloromethane-methanol volume ratio of 40:1 to 1:1, collecting the elution component, and carrying out Toyopearl column chromatography purification to obtain the imidazole dimer alkaloid shown in the formula III.
6. The process for producing an imidazole dimer alkaloid according to claim 5, wherein in step (2), the concentration of the aqueous ethanol solution used is 95%.
7. The process for producing an imidazole dimer alkaloid according to claim 5 or 6, wherein in step (3), the aqueous tartaric acid solution is used at a concentration of 2% and the aqueous sodium hydroxide solution is used at a concentration of 1-2%.
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| Brick dark tea: a review of the manufacture, chemical constituents and bioconversion of the major chemical components during fermentation;Wen-Jun Zheng et al.;《Phytochem Rev》;20150328;第14卷;第499-523页 * |
| Decomposition and Stabilization of Drugs. X. A New Decomposition Route of Caffeine in Alkaline Solution;KAZUO KIGASAWA et al.;《Chem. Pharm. Bull.》;19741231;第22卷(第10期);第2248-2451页 * |
| Imidazole, oxazole and thiazole alkaloids;Zhong Jin;《Natural Product Reports》;20060328;第22卷;第196-229页 * |
| 海洋二聚吡咯-咪唑类生物碱;王珏等;《化学进展》;20111031;第23卷(第10期);第2065-2078页 * |
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