CN114716432A - Crystal form of demethylenetetrahydroberberine hydrochloride and preparation method thereof - Google Patents

Crystal form of demethylenetetrahydroberberine hydrochloride and preparation method thereof Download PDF

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CN114716432A
CN114716432A CN202210092386.8A CN202210092386A CN114716432A CN 114716432 A CN114716432 A CN 114716432A CN 202210092386 A CN202210092386 A CN 202210092386A CN 114716432 A CN114716432 A CN 114716432A
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demethylenetetrahydroberberine
hydrochloride
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张玉彬
张元强
闻婧
朱倩倩
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China Pharmaceutical University
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Abstract

The invention belongs to the technical field of medicinal chemistry, and particularly relates to three crystal forms A, B, C of demethylenetetrahydroberberine hydrochloride and a preparation method thereof. The diffraction characteristic diffraction peaks of the crystal form X-ray powder diffraction are about 6.264, 9.603, 10.622, 11.933, 12.338, 13.206, 15.011, 15.409, 17.879, 18.273, 18.569, 18.844, 19.223, 19.865, 20.174, 21.348, 21.742, 22.393, 23.280, 24.699, 26.133, 26.700, 27.180, 27.630, 28.419, 29.088 and 31.801 degrees. The invention also relates to a preparation method of the three crystal forms of the demethylenetetrahydroberberine hydrochloride. The preparation method has the advantages of simple process, high yield and low cost; the product has high purity and stable quality.

Description

Crystal form of demethylenetetrahydroberberine hydrochloride and preparation method thereof
The application is application number: 2020106510592, filing date: 2020.07.08, the invention relates to a crystal form of hydrochloric acid demethylenetetrahydroberberine and a divisional application of a preparation method thereof.
Technical Field
The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to three crystal forms of demethylenetetrahydroberberine hydrochloride and a preparation method thereof.
Background
Demethylenetetrahydroberberine hydrochloride, english name: demethylethyltetrahydroberine Hydrochloride, molecular formula: c19H22NO4Cl, molecular weight: 363.7. the organic structure part of the berberine is named as: 6H-dibenzo [ a, g ]]Quinolizine-2, 3-diol, 5,8,13,13 a-tetrahydro-9, 10-dimethoxy. The structural formula of the demethylenetetrahydroberberine hydrochloride is shown as a formula (I).
Figure BDA0003489677840000011
The patent reports that the demethylenetetrahydroberberine hydrochloride has low toxicity, anti-inflammatory, antioxidant and other biological activities. And has stronger treatment potential in the fields of alcoholic liver injury, non-alcoholic fatty liver, thioacetamide induced chemical liver injury and the like, and is a potential liver protection medicament. In addition, there is an article reporting that demethylenetetrahydroberberine hydrochloride has an inhibitory activity against tissue factor procoagulant. In conclusion, the application of the demethyleneberberine hydrochloride provides a choice for treating clinical diseases.
Tetrahydrobererine (THB) is derived from tuber of corydalis tuber of Papaveraceae, belongs to tetrahydroisoquinoline alkaloid, and can also be hydrogenated from berberine, and has various biological activities. The demethylenetetrahydroberberine can be obtained by metabolizing or chemically modifying tetrahydroberberine. Because the content of the hydrochloric acid demethylenetetrahydroberberine in natural plants is low, the extraction cost is high, and the price is high. The chemical synthesis method is used for preparing the demethylenetetrahydroberberine hydrochloride, and the chemical synthesis method comprises the steps of using cheap berberine as a raw material, utilizing reducing agents such as sodium borohydride and the like to reduce the berberine to obtain the tetrahydroberberine, removing methylene from the tetrahydroberberine through a sulfuric acid-phloroglucinol system to obtain the demethylenetetrahydroberberine hydrochloride, and recrystallizing to obtain the pure demethylenetetrahydroberberine hydrochloride.
The method obtains a stable and high-purity product by using a recrystallization method, improves the related synthesis process, reduces the synthesis cost of the demethylenetetrahydroberberine hydrochloride, and improves the yield of the demethylenetetrahydroberberine hydrochloride. At present, no patent report in literature relates to the preparation of crystal form of demethylenetetrahydroberberine hydrochloride, but the crystal form structure of the medicament is extremely important to the stability, dissolution and bioavailability of the medicament, and the proper crystal form of the medicament is very important to pharmaceutical research. In order to further research the pharmacological activity of the demethylenetetrahydroberberine hydrochloride, the research of the crystal form of the demethylenetetrahydroberberine hydrochloride is necessary.
Disclosure of Invention
The invention aims to provide three crystal forms of demethylenetetrahydroberberine hydrochloride and preparation methods thereof.
A crystal form of demethylenetetrahydroberberine hydrochloride, which is named as type A, is characterized in that: characteristic diffraction peaks in an X-ray powder diffraction spectrum expressed by Cu-Kalpha radiation and a2 theta +/-0.2 DEG diffraction angle are 6.264, 9.603, 10.622, 11.933, 12.338, 13.206, 15.011, 15.409, 17.879, 18.273, 18.569, 18.844, 19.223, 19.865, 20.174, 21.348, 21.742, 22.393, 23.280, 24.699, 26.133, 26.700, 27.180, 27.630, 28.419, 29.088 and 31.801 DEG; the DSC graph contains 1 heat absorption peak at 258.1 +/-3 ℃; the infrared absorption characteristic peak is as follows: 422.3cm-1、451.2cm-1、513.8cm-1、530.7cm-1、554.3cm-1、631.7cm-1、677.6cm-1、694.2cm-1、732.7cm-1、793.7cm-1、810.0cm-1、865.3cm-1、882.9cm-1、903.8cm-1、927.9cm-1、957.8cm-1、983.5cm-1、1008.2cm-1、1040.1cm-1、1092.9cm-1、1115.2cm-1、1161.5cm-1、1188.1cm-1、1232.0cm-1、1279.7cm-1、1302.1cm-1、1318.5cm-1、1333.8cm-1、1358.3cm-1、1373.4cm-1、1391.9cm-1、1426.6cm-1、1443.7cm-1、1501.3cm-1、1524.0cm-1、1587.1cm-1、1605.8cm-1、1622.3cm-1、2402.8cm-1、2458.7cm-1、2585.4cm-1、2668.0cm-1、2689.3cm-1、2689.3cm-1、2791.9cm-1、2833.3cm-1、2908.4cm-1、2937.5cm-1、2976.8cm-1、2995.5cm-1、3019.7cm-1、3142.2cm-1、3293.4cm-1。
A crystal form of demethylenetetrahydroberberine hydrochloride, named as form B, is characterized in that: the characteristic diffraction peak in an X-ray powder diffraction spectrum expressed by Cu-Ka radiation and 2 theta +/-0.2 DEG diffraction angle is as follows: 6.299, 10.644, 12.292, 12.559, 13.236, 15.034, 16.230, 18.604, 18.943, 19.912, 20.199, 21.381, 23.302, 24.738, 26.146, 27.137, 28.457, 29.106, 30.726, 31.862, 32.413, 36.198, 38.462 °; the DSC spectrum contains 1 endothermic peak, and the peak position is 253.7 +/-3 ℃; the infrared characteristic absorption peak is: 422.0cm-1、450.2cm-1、506.2cm-1、530.4cm-1、550.7cm-1、631.9cm-1、678.1cm-1、701.6cm-1、733.5cm-1、793.2cm-1、808.4cm-1、864.3cm-1、882.7cm-1、904.0cm-1、923.8cm-1、948.5cm-1、958.8cm-1、982.3cm-1、1008.5cm-1、1043.3cm-1、1094.1cm-1、1115.4cm-1、1160.8cm-1、1187.7cm-1、1230.9cm-1、1286.8cm-1、1319.5cm-1、1333.1cm-1、1358.2cm-1、1373.7cm-1、1391.1cm-1、1424.8cm-1、1446.6cm-1、1502.8cm-1、1523.8cm-1、1586.8cm-1、1606.8cm-1、1624.0cm-1、2403.1cm-1、2457.8cm-1、2589.9cm-1、2670.0cm-1、2691.6cm-1。
A crystal form of demethylenetetrahydroberberine hydrochloride, named as form C, is characterized in that: the characteristic diffraction peak in an X-ray powder diffraction spectrum expressed by Cu-Ka radiation and 2 theta +/-0.2 DEG diffraction angle is as follows: 9.722, 12.030, 12.452, 15.555, 17.986, 18.366, 19.367, 20.328, 21.867, 22.501, 23.712, 24.090, 24.996, 26.226, 26.840, 27.747, 28.103, 28.521, 29.095, 30.944, 32.199, 33.527, 33.943, 38.700 °; the DSC graph contains 1 endothermic peak, and the peak position is 250.8 plus or minus 3 ℃; the infrared characteristic absorption peak is: 424.0cm-1、453.1cm-1、514.8cm-1、556.3cm-1、613.1cm-1、633.0cm-1、652.2cm-1、678.2cm-1、692.3cm-1、733.1cm-1、741.4cm-1、758.3cm-1、797.8cm-1、811.5cm-1、867.3cm-1、883.1cm-1、904.0cm-1、928.9cm-1、957.6cm-1、986.0cm-1、1007.7cm-1、1037.2cm-1、1057.5cm-1、1091.8cm-1、1116.5cm-1、1164.8cm-1、1189.2cm-1、1221.5cm-1、1234.7cm-1、1278.0cm-1、1315.8cm-1、1334.8cm-1、1349.9cm-1、1374.1cm-1、1393.5cm-1、1429.6cm-1、1442.1cm-1、1459.7cm-1、1495.4cm-1、1524.8cm-1、1587.4cm-1、1605.0cm-1、1621.7cm-1、2402.3cm-1、2469.3cm-1、2572.5cm-1、2657.1cm-1、2684.2cm-1、2798.7cm-1、2836.2cm-1、2906.4cm-1、2941.1cm-1、2961.3cm-1、2999.0cm-1、3142.5cm-1、3298.3cm-1、3518.5cm-1。
A method for preparing the crystal form of the demethylenetetrahydroberberine hydrochloride is characterized in that: the method is realized by the following steps:
(1) the hydrochloric acid tetrahydroberberine passes through a sulfuric acid-phloroglucinol system to obtain hydrochloric acid demethylenetetrahydroberberine;
(2) dissolving the demethylenetetrahydroberberine hydrochloride obtained in the step (1) in water, cooling and crystallizing to obtain a demethylenetetrahydroberberine hydrochloride A crystal form;
(3) dissolving the demethylenetetrahydroberberine hydrochloride obtained in the step (1) by using ethanol or methanol, and recrystallizing to obtain a crystal form B of the demethylenetetrahydroberberine hydrochloride;
(4) dissolving the demethylenetetrahydroberberine hydrochloride obtained by mixing in the step (1) by using ethanol or methanol, adding ethyl acetate, and recrystallizing at 4 ℃ to obtain a demethylenetetrahydroberberine hydrochloride C crystal form;
the preparation method of the crystal form of the hydrochloric acid demethylenetetrahydroberberine comprises the step (2) of adding a small amount of seed crystals.
The preparation method of the crystal form of the hydrochloric acid demethylenetetrahydroberberine comprises the step (4) of adding a small amount of seed crystals, adding 10-20% of diethyl ether by volume, adding 10-20% of dimethylbenzene by volume and adding 10-20% of methylbenzene by volume.
The crystal form A, B, C of the hydrochloric acid demethylenetetrahydroberberine is characterized in that the medicine composition of the crystal form of the hydrochloric acid demethylenetetrahydroberberine can be tablets, capsules, pills, injections, sustained-release agents and various particle drug delivery systems.
The three crystal form drug combinations of the demethylenetetrahydroberberine hydrochloride can be tablets, capsules, pills, injections, sustained-release agents and various particle drug delivery systems.
The invention has the beneficial technical effects that: the crystal form of the demethylenetetrahydroberberine hydrochloride provided by the invention has better stability. The preparation method has the advantages of simple process, high yield, good product quality and low preparation cost.
Drawings
FIG. 1 is X-ray powder diffractogram of A crystal form of demethylenetetrahydroberberine hydrochloride of the present invention;
FIG. 2 is a differential scanning calorimetry chart of crystalline form A of demethylenetetrahydroberberine hydrochloride;
FIG. 3 is an infrared spectrum of a crystal form A of demethylenetetrahydroberberine hydrochloride;
FIG. 4 is a thermogram of crystalline form A of demethylenetetrahydroberberine hydrochloride;
FIG. 5 is a powder diffractogram of crystal form B of demethylenetetrahydroberberine hydrochloride of the present invention;
FIG. 6 is a differential scanning calorimetry chart of crystalline form B of demethylenetetrahydroberberine hydrochloride;
FIG. 7 is an infrared spectrum of crystalline form B of demethylenetetrahydroberberine hydrochloride;
FIG. 8 is a thermogram of crystalline form B of demethylenetetrahydroberberine hydrochloride;
FIG. 9 is an X-ray powder diffractogram of crystalline form C of demethylenetetrahydroberberine hydrochloride of the present invention;
FIG. 10 is a differential scanning calorimetry trace of crystalline form C of demethylenetetrahydroberberine hydrochloride;
FIG. 11 is an infrared spectrum of crystalline form C of demethylenetetrahydroberberine hydrochloride;
FIG. 12 is a thermogram of crystalline form C of demethylenetetrahydroberberine hydrochloride;
FIG. 13 is an HPLC check chart of demethylenetetrahydroberberine hydrochloride;
FIG. 14 is a mass spectrum of demethylenetetrahydroberberine hydrochloride;
FIG. 15 is a hydrogen spectrum of demethylenetetrahydroberberine hydrochloride;
FIG. 16 is a dissolution curve of demethylenetetrahydroberberine hydrochloride A, B, C crystal form water system;
FIG. 17 is a dissolution curve of demethylenetetrahydroberberine hydrochloride A, B, C crystal form ethanol system.
FIG. 18 shows the blood concentration detection of the crystalline form A, B, C of demethylenetetrahydroberberine hydrochloride
Detailed Description
The following examples may assist those skilled in the art in a more complete understanding of the present invention, but are not intended to limit the invention in any way.
The structural characterization of the crystal form of the demethylenetetrahydroberberine hydrochloride adopts X-ray powder diffraction, differential scanning calorimetry, infrared spectrum and thermogravimetric analysis.
In the sulfuric acid-phloroglucinol system, the temperature range is as follows: 60-100 ℃, sulfuric acid concentration range: 40-80%, and the proportion range of phloroglucinol and tetrahydroberberine is as follows: 1:3-3:1.
Example 1 Synthesis of crystalline form A of Desmethylenetetrahydroberberine hydrochloride
Adding berberine hydrochloride 5g into ethanol 50ml, stirring, slowly adding sodium borohydride 0.5g, reacting for 4h, and recovering tetrahydroberberine hydrochloride after reaction. Adding tetrahydroberberine hydrochloride into phloroglucinol-H2SO4Stirring in the system to completely dissolve, reacting for 0.5h under the catalysis of phloroglucinol, and removing methylene to obtain demethylenetetrahydroberberine hydrochloride. Dissolving the obtained product in water, recrystallizing, filtering, collecting filter cake, drying to obtain demethylenetetrahydroberberine hydrochloride A crystal, weighing, and detecting. The obtained solid has X-ray powder diffraction pattern as shown in figure 1, DSC pattern as shown in figure 2, infrared spectrum as shown in figure 3, and thermogravimetric analysis pattern as shown in figure 4.
Example 2 Synthesis of crystalline form B of demethylenetetrahydroberberine hydrochloride
Adding berberine hydrochloride 5g into ethanol 50ml, stirring, slowly adding sodium borohydride 0.5g, reacting for 4h, and collecting tetrahydroberberine hydrochloride after reaction. Adding tetrahydroberberine hydrochloride into phloroglucinol-H2SO4Stirring in the system to completely dissolve, reacting for 0.5h under the catalysis of phloroglucinol, and removing methylene to obtain demethylenetetrahydroberberine hydrochloride. Dissolving the obtained product in methanol, adding ethyl acetate, recrystallizing, filtering, collecting filter cake, drying to obtain demethylenetetrahydroberberine hydrochloride B crystal, weighing, and detecting. The obtained solid has X-ray powder diffraction pattern as shown in FIG. 5, DSC pattern as shown in FIG. 6, infrared spectrum as shown in FIG. 7, and thermogravimetric analysis as shown in FIG. 8.
Example 3 Synthesis of crystalline form C of Demethylenetetrahydroberberine hydrochloride
Adding berberine hydrochloride 5g into ethanol 50ml, stirring, slowly adding sodium borohydride 0.5g, reacting for 4h, and collecting tetrahydroberberine hydrochloride after reaction. Adding tetrahydroberberine hydrochloride into phloroglucinol-H2SO4Stirring in the system to completely dissolve, reacting for 0.5h under the catalysis of phloroglucinol, and removing methylene to obtain demethylenetetrahydroberberine hydrochloride. Dissolving the obtained product with ethanol, adding ethyl acetate, recrystallizing overnight at 4 deg.C, vacuum filtering, collecting filter cake, drying to obtain demethylenetetrahydroberberine hydrochloride C crystal, weighing, and detecting. The obtained solid has X-ray powder diffraction pattern as shown in FIG. 9, DSC pattern as shown in FIG. 10, infrared spectrum as shown in FIG. 11, and thermogravimetric analysis as shown in FIG. 12.
Example 4 HPLC detection of demethylenetetrahydroberberine hydrochloride
The method comprises the following steps: using HPLC method to analyze, dissolving the hydrochloric acid demethylenetetrahydroberberine in methanol, filtering the result, and finally preparing a sample with the final concentration of 20 ng/mu l for analysis. HPLC chromatograph: agilent 1100 hplc. A chromatographic column: agilent Eclipse XDB-C18(4.6X150mm, 5 μm). The mobile phase comprises (A)20mM phosphate buffer salt solution and (B) acetonitrile. HPLC gradient elution conditions of 0min, 20% B; 0-13min, 55% B; 13-14min, 55% B; 14-15min, 20% B; 15-16min, 20% B. The detection wavelength is 280nm, the speed is 1.0ml/min, and the injection volume is 5 mul.
As a result: the HPLC analysis result is shown in FIG. 13, the retention time is 3.817min, and the purity is more than 98.0%.
Example 5 identification and detection of the structure of Desmethylenetetrahydroberberine hydrochloride
The method comprises the following steps: the molecular structure identification adopts mass spectrum and nuclear magnetic resonance spectrum analysis. The mass spectrometer is a Waters Q-TOF MicroTM, and the ionization mode is as follows: ESI (+), mass scan range: m/z 80-1000, capillary voltage: 2500v, cone voltage: 25v, ion source temperature: 100 ℃, atomization temperature: 200 ℃, cone orifice air flow rate: 50L/hr, atomizing gas flow rate: 400L/hr. The nuclear magnetic resonance spectrometer (1H-NMR) is AVANCE AV-300, the irradiation frequency is 300MHz, and the used solvents are: deuterated dimethyl sulfoxide (DMSO-d6) and Tetramethylsilane (TMS) as a standard.
As a result, the results of the positive ion mass spectrometry of demethylenetetrahydroberberine are shown in FIG. 14, ESI-MS (M/z) molecular ion group peak [ M-CI ]]327.1 and [ MH-Cl]+328.1. This is consistent with the theoretical molecular weight of demethylenetetrahydroberberine, the chloride ion in demethylenetetrahydroberberine hydrochloride is not shown in mass spectrum ESI (+). Identifying chloride ions according to the identification (1) reaction method of related chlorides in Chinese pharmacopoeia. 0.1 g of demethylenetetrahydroberberine hydrochloride is taken, 10ml of water is added, after the solution is slowly heated and dissolved, 0.5ml of nitric acid is added, the solution is cooled and placed for 10 minutes, the solution is filtered, after silver nitrate test solution is dripped into the filtrate, white creamy sediment is generated, the sediment is centrifuged, the sediment is collected, ammonia test solution is added into the sediment for dissolving, and after diluted nitric acid is added for acidification, the sediment is generated. The experiment sample shows that the hydrochloric acid demethylenetetrahydroberberine contains chloride ions. Mass spectrometry and chemical analysis show that the molecular weight of the hydrochloric acid demethylenetetrahydroberberine is correct.
Method for preparing demethylenetetrahydroberberine hydrochloridelH-NMR (DMSO-d6) Hydrogen Spectroscopy results are shown in FIG. 15, 1H NMR (DMSO-d6,300MHz) d:2.79(m,1H,6-H),3.04(m,1H,13-H),3.23(m,1H,5-H),3.41(m,2H,6-H,5-H),3.58(m,1H,13-H),3.77(m,1H,8-H),3.79(s,3H, -OCH 3),3.82(s,3H, -OCH 3),4.35(m,1H,14-H),4.59(m,1H,8-H),6.60(s,1H,4-H),6.79(s,1H,1-H),7.04(d, J ═ 8.5Hz,1H,12-H),7.07(d, J ═ 8H, H ═ H), 11H, — 9H (s, 9 Hz), 9.9H, — H (s, 9 Hz), (S, H, 9 Hz), (S, H, 9-H, 9 Hz), (S, H, 1H, -OH).
Example 6 stability test
A proper amount of demethylenetetrahydroberberine hydrochloride A, B and C crystals are respectively placed in three parts of dishes with numbers of A1, A2, A3, B1, B2, B3, C1, C2 and C3, and are respectively placed in the following conditions (storage condition 1: high temperature at 60 ℃, storage condition 2: relative humidity 92.5 high humidity, storage condition 3: 4500lx +/-500 lx) for stability experiments. The results of the experiments are shown in tables 1, 2 and 3.
TABLE 1 high temperature experimental stability study (60 + -2 ℃ C.) of demethylenetetrahydroberberine hydrochloride crystals
Figure BDA0003489677840000051
Figure BDA0003489677840000061
TABLE 2 study of stability of high humidity test of demethylenetetrahydroberberine hydrochloride crystals (RH90 + -5%)
Figure BDA0003489677840000062
TABLE 3 stability study of strong light irradiation experiment of demethylenetetrahydroberberine hydrochloride crystals (RH90 + -5%)
Figure BDA0003489677840000063
And (4) conclusion: in the experimental study of high-temperature stability, the demethylenetetrahydroberberine hydrochloride B crystal shows better stability, and the demethylenetetrahydroberberine hydrochloride C crystal has poorer stability. In the experimental study of high humidity stability, the demethylenetetrahydroberberine hydrochloride C crystal shows better stability, and the demethylenetetrahydroberberine hydrochloride A crystal has poorer stability. In the experimental study of strong light irradiation stability, the demethylenetetrahydroberberine hydrochloride B crystal shows better stability, and the demethylenetetrahydroberberine hydrochloride A crystal and the demethylenetetrahydroberberine hydrochloride C crystal have poorer stability.
Example 7 solubility study
Taking a proper amount of the demethylenetetrahydroberberine hydrochloride A, B and C crystals respectively, and researching the solubility of the demethylenetetrahydroberberine hydrochloride A, B and C crystals at different temperatures by using water and ethanol as solvents.
1. Solubility research of hydrochloric acid demethylenetetrahydroberberine A, B and C crystals in water system
Taking a proper amount of the demethylenetetrahydroberberine hydrochloride crystals A, B and C respectively, and researching the solubility of the demethylenetetrahydroberberine hydrochloride crystals A, B and C at different temperatures in a water system. 20ul samples were taken at each temperature node and tested by HPLC, and the results are shown in FIG. 16.
And (4) conclusion: the solubility of the hydrochloric acid demethylenetetrahydroberberine A, B and C crystals in an aqueous phase system is slowly increased along with the increase of the temperature. The solubility of the demethylenetetrahydroberberine hydrochloride C crystal is the best, and the solubility of the demethylenetetrahydroberberine hydrochloride B crystal is the worst.
2. Solubility research of hydrochloric acid demethylenetetrahydroberberine A, B and C crystals in ethanol system
Taking a proper amount of the demethylenetetrahydroberberine hydrochloride crystals A, B and C respectively, and researching the solubility of the demethylenetetrahydroberberine hydrochloride crystals A, B and C at different temperatures under an ethanol system. 20ul samples were taken at each temperature node and tested by HPLC, and the results are shown in FIG. 17.
And (4) conclusion: the solubility of the hydrochloric acid demethylenetetrahydroberberine A, B and C crystals in an ethanol system is increased along with the increase of the temperature. The solubility of the demethylenetetrahydroberberine hydrochloride A crystal is the best, and the solubility of the demethylenetetrahydroberberine hydrochloride B crystal is the worst.
Example 8 measurement of blood levels of crystalline Desmethylenetetrahydroberberine hydrochloride A, B, C
The male SD rats are adaptively raised for 3 days, and rats with the body weight (220 +/-20) g are selected to be divided into 4 groups, namely a blank group, a demethylenetetrahydroberberine hydrochloride A crystal form group, a demethylenetetrahydroberberine hydrochloride B crystal form group and a demethylenetetrahydroberberine hydrochloride C crystal form group. Before the experiment, the stomach is fasted for 12h and emptied, and the dosage of the ig administration of rats in the experimental group is 300 mg/kg. Respectively administering to rat ig for 5min, 15min, 30min, 1h, 2h, 6h, 12h, collecting blood 0.2ml from orbit, centrifuging whole blood, collecting plasma 100 μ l, and freezing at-20 deg.C. Taking 100 μ l plasma sample, adding 200 μ l methanol, vortex for 5min, centrifuging at 10000r/min for 15min in a centrifuge, collecting supernatant, detecting by HPLC, the experimental result is shown in figure 18, and the hydrochloric acid demethylenetetrahydroberberine A crystal form Cmax＝19.958μg×ml-1，Tmax2 h; crystal form C of demethylenetetrahydroberberine hydrochloride Bmax＝10.38μg×ml-1，Tmax0.5 h; crystal form C of demethylenetetrahydroberberine hydrochloridemax＝55.352μg×ml-1，Tmax0.25 h. Calculating A according to the experimental resultAUC＝91.67258μg×ml-1×h＞CAUC＝47.50667μg×ml-1×h＞BAUC19.2071 μ g × ml-1 × h. The absorption of the hydrochloric acid demethylenetetrahydroberberine A, C crystal form is obviously better than that of hydrochloric acid demethylenetetrahydroberberine B crystal form.

Claims (4)

1. A crystal form of demethylenetetrahydroberberine hydrochloride is characterized in that: the crystal form is named as B form, and the characteristic diffraction peaks in an X-ray powder diffraction spectrum expressed by Cu-Kalpha radiation and 2 theta +/-0.2 degrees of diffraction angles are as follows: 6.299, 10.644, 12.292, 12.559, 13.236, 15.034, 16.230, 18.604, 18.943, 19.912, 20.199, 21.381, 23.302, 24.738, 26.146, 27.137, 28.457, 29.106, 30.726, 31.862, 32.413, 36.198, 38.462 °; the DSC spectrum contains 1 endothermic peak, and the peak position is 253.7 +/-3 ℃; the infrared characteristic absorption peak is: 422.0cm-1、450.2 cm-1、506.2 cm-1、530.4 cm-1、550.7cm-1、631.9cm-1、678.1cm-1、701.6cm-1、733.5cm-1、793.2cm-1、808.4cm-1、864.3cm-1、882.7cm-1、904.0cm-1、923.8cm-1、948.5cm-1、958.8cm-1、982.3cm-1、1008.5cm-1、1043.3cm-1、1094.1cm-1、1115.4cm-1、1160.8cm-1、1187.7cm-1、1230.9 cm-1、1286.8 cm-1、1319.5 cm-1、1333.1 cm-1、1358.2 cm-1、1373.7 cm-1、1391.1 cm-1、1424.8 cm-1、1446.6 cm-1、1502.8 cm-1、1523.8 cm-1、1586.8 cm-1、1606.8 cm-1、1624.0 cm-1、2403.1 cm-1、2457.8 cm-1、2589.9 cm-1、2670.0 cm-1、2691.6 cm-1。
2. A process for preparing the crystalline form of demethylenetetrahydroberberine hydrochloride of claim 1, characterized in that: the method is realized by the following steps:
(1) the hydrochloric acid tetrahydroberberine passes through a sulfuric acid-phloroglucinol system to obtain hydrochloric acid demethylenetetrahydroberberine;
(2) dissolving the demethylenetetrahydroberberine hydrochloride obtained in the step (1) by using ethanol or methanol, and recrystallizing to obtain a demethylenetetrahydroberberine hydrochloride B crystal form.
3. The preparation method of the crystalline form of demethylenetetrahydroberberine hydrochloride according to claim 2, wherein: the sulfuric acid-phloroglucinol system in the step (1) is as follows: the temperature is 60-100 ℃, the concentration of the sulfuric acid is 40-80%, and the mass ratio of the phloroglucinol to the tetrahydroberberine is 1:3-3: 1.
4. The pharmaceutical composition containing the crystalline form of demethylenetetrahydroberberine hydrochloride according to claim 1, wherein the pharmaceutical composition is in the form of a tablet, a capsule, a pill, an injection or a sustained release formulation.
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CN109432096A (en) * 2018-11-12 2019-03-08 中国药科大学 Hydrochloric acid goes application of the methylene N-1 in preparation prevention or treatment alcoholic liver disease and non-alcoholic fatty liver disease drug

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WO2009007457A2 (en) * 2007-07-12 2009-01-15 Exonhit Therapeutics Sa Compounds and methods for modulating rho gtpases
CN108033958A (en) * 2018-02-06 2018-05-15 中国药科大学 Demethyleneberberinehydrochloride hydrochloride crystal form and preparation method thereof
CN109293655A (en) * 2018-02-06 2019-02-01 中国药科大学 Demethyleneberberinehydrochloride hydrochloride crystal form and preparation method thereof
CN109432096A (en) * 2018-11-12 2019-03-08 中国药科大学 Hydrochloric acid goes application of the methylene N-1 in preparation prevention or treatment alcoholic liver disease and non-alcoholic fatty liver disease drug

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