WO2017133465A1 - 一种黄芩苷镁化合物及其制备方法与它的用途 - Google Patents

一种黄芩苷镁化合物及其制备方法与它的用途 Download PDF

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WO2017133465A1
WO2017133465A1 PCT/CN2017/071616 CN2017071616W WO2017133465A1 WO 2017133465 A1 WO2017133465 A1 WO 2017133465A1 CN 2017071616 W CN2017071616 W CN 2017071616W WO 2017133465 A1 WO2017133465 A1 WO 2017133465A1
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baicalin
magnesium
preparation
compound
water
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PCT/CN2017/071616
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English (en)
French (fr)
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刘翠哲
王志轩
苗策禹
刘丽艳
薛禾菲
刘喜纲
杜闻杉
刘一鑫
张琳
毛晓霞
刘进军
赵桂琴
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承德医学院
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Priority to JP2018558469A priority Critical patent/JP2019509332A/ja
Priority to EP17746785.9A priority patent/EP3412679B1/en
Publication of WO2017133465A1 publication Critical patent/WO2017133465A1/zh
Priority to US16/052,419 priority patent/US10196417B2/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H23/00—Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
    • C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
    • C07H17/06—Benzopyran radicals
    • C07H17/065—Benzo[b]pyrans
    • C07H17/07—Benzo[b]pyran-4-ones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A61P29/02—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] without antiinflammatory effect
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives
    • C07H1/06—Separation; Purification
    • C07H1/08—Separation; Purification from natural products

Definitions

  • the invention belongs to the field of medical technology. More specifically, the present invention relates to a baicalin compound, and to a method for extracting and preparing the baicalin compound, and to the use of the baicalin compound.
  • Huang Wei was first published in Shennong Materia Medica. It is the dry root of Scutellaria baicalensis Georgi, a plant of the Labiatae family. It has the functions of clearing away heat and dampness, purging fire and detoxification, and stopping bleeding.
  • Baicalin is a kind of flavonoid compound contained in Astragalus membranaceus. Its molecular formula is C 21 H 18 O 11 , and its content in Astragalus is more than 9% by weight, even up to 20% by weight. It is one of the main active ingredients of Astragalus.
  • baicalin has the functions of scavenging oxygen free radicals, antiarrhythmia, dilating cardiovascular and cerebrovascular, antihypertensive sedation, protecting liver and gallbladder, and resisting pathogens.
  • preparations such as baicalin tablets and baicalin capsules, which are mainly used for treating acute and chronic hepatitis and cardiovascular diseases.
  • Magnesium is an essential element involved in the normal life activities and metabolic processes of living organisms. Magnesium affects many biological functions of cells, affects potassium and calcium transport, regulates signal transmission, participates in energy metabolism, protein and nucleic acid synthesis, catalyzes enzyme activation and inhibition, and regulates cell cycle, cell proliferation and cell differentiation; It is also involved in maintaining the stability of the genome and is also involved in oxidative stress and tumorigenesis in the body.
  • Baicalin can be dissolved in alkaline solution such as sodium hydroxide, sodium carbonate or sodium bicarbonate, but it is unstable in lye, it will gradually dark brown, slightly soluble in hot glacial acetic acid, and insoluble in methanol, ethanol and C. Ketone, insoluble in water, benzene, ether, chloroform. Because baicalin is almost insoluble in water, oral bioavailability is low, which greatly limits its wide clinical application.
  • baicalin mainly involving the preparation of baicalin into metal complexes, ⁇ -cyclodextrin inclusion complexes, solid dispersions, phospholipid complexes, microparticles, nanometers.
  • baicalin is prepared by this method.
  • the original form of baicalin in Astragalus is a magnesium salt form. This form has good water solubility and can be extracted by water dissolution.
  • acid precipitation destroys its original structure, which makes the water solubility of baicalin worse. The speed is slow, the bioavailability is low, and the efficacy of baicalin is also reduced.
  • the invention finds a method for extracting baicalin magnesium compound from Astragalus membranaceus without changing the original existence mode of baicalin in Astragalus, and also finds a method for synthesizing baicalin magnesium compound by using commercially available baicalin as raw material, Restored the existence of baicalin in Astragalus.
  • the baicalin magnesium compound can be used as a substitute for baicalin, and the solubility of baicalin is greatly improved, the absorption rate of baicalin is increased, the oral bioavailability is improved, and a water-soluble injection or powder injection can be conveniently prepared.
  • the compound can also exert synergistic pharmacological effects of baicalin and magnesium ions to increase the efficacy.
  • the present inventors have finally completed the present invention on the basis of summarizing the prior art and through a large number of experimental research and analysis.
  • Another object of the present invention is to provide a process for the preparation of the baicalin magnesium compound.
  • Another object of the invention is to provide the use of the baicalin compound.
  • the present invention has been achieved by the following technical solutions.
  • the present invention relates to a baicalin magnesium compound.
  • the baicalin compound is a compound having the following chemical formula (I):
  • the invention also relates to a method for preparing a baicalin magnesium compound.
  • the baicalin powder was added to purified water in a ratio of 1:20 to 100 in terms of grams of purified baicalin in milliliters, and uniformly mixed to obtain a baicalin suspension.
  • the magnesium compound is added to the suspension of baicalin obtained in step A, and uniformly mixed to obtain a suspension of baicalin containing magnesium ions;
  • the magnesium ion-containing baicalin suspension obtained in the step B is reacted at a temperature of 20 to 70 ° C until the reaction system becomes clear and transparent, and filtered to obtain a filtrate;
  • the filtrate obtained in the step C is dried to obtain the baicalin magnesium compound.
  • the magnesium compound is selected from the group consisting of magnesium hydroxide, magnesium oxide, basic magnesium carbonate, magnesium acetate, magnesium sulfate, magnesium nitrate or magnesium chloride.
  • the invention further relates to a process for the preparation of another baicalin magnesium compound.
  • the macroporous adsorption resin is immersed in a 95% ethanol aqueous solution at a concentration for 20 to 28 hours, and then packed in a wet manner, followed by a concentration of 95% ethanol aqueous solution at a volume of 1.5 to 2.5 BV/h.
  • the flow rate is passed through the macroporous adsorption resin column until the effluent and water are mixed at a volume ratio of 1:5, and then no longer appear white turbid, and then eluted with distilled water at a flow rate of 1.5 to 2.5 BV/h to be colorless;
  • the ratio of xanthine to ethanol in ml the ratio of 1:8 to 15 is added to the concentration of 40-60% ethanol solution by volume, and the mixture is extracted at a temperature of 55-65 ° C under stirring conditions. 1.2 hours, then separated, to obtain an extract and a residue; the residue was repeatedly extracted 2 to 3 times under the same conditions as above, and the residue was discarded; the extract was combined to obtain a jaundice Extract
  • Step B Let the Astragalus extract obtained in Step B be adsorbed by the large pore adsorption resin column pretreated in Step A at a flow rate of 1.5 to 2.5 BV/h, and then washed with 4 to 6 large pore adsorption resin bed volume water, and the washing water is discarded; Further, using 4 to 6 macroporous adsorption resin bed volume concentrations to elute with 45 to 55% by volume of aqueous ethanol solution, and collecting the eluate containing baicalin;
  • the eluate containing baicalin obtained in the step C is concentrated under reduced pressure at a temperature of 35 to 65 ° C until the volume of the concentrate is 1 to 15 times the weight of the astragalus medicinal material, and then dried to obtain a jaundice.
  • the crude extract of the baicalin magnesium compound is purified in step D to obtain the baicalin magnesium compound.
  • the macroporous adsorption resin is selected from the group consisting of HPD-100, AB-8, D101 or YWD06B.
  • the macroporous adsorption resin has a particle size of 10 to 80 mesh.
  • the ratio of the diameter of the macroporous adsorption resin column to the column height is 1:3-8.
  • the drying is by heat drying, spray drying or freeze drying.
  • the obtained baicalin compound is purified by recrystallization, octadecylsilane reverse phase column, preparative liquid chromatography to achieve a content of baicalin magnesium compound of 95 by weight. %the above.
  • the invention further relates to the use of said baicalin magnesium compound.
  • the baicalin compound replaces astragalus or baicalin in a single or compound preparation of traditional Chinese medicine containing xanthine or baicalin in an equal dose, or it is prepared for treating liver injury, cerebral ischemia, diabetes, inflammation or tumor drug Use in. It is also very convenient to prepare a water-soluble injection or powder injection.
  • the present invention relates to a baicalin magnesium compound.
  • the baicalin compound is a compound having the following chemical formula (I):
  • the invention also relates to a method for preparing a baicalin magnesium compound.
  • the invention provides a method for extracting baicalin magnesium compound from Astragalus membranaceus, and also provides a method for synthesizing baicalin magnesium compound by using commercially available baicalin as raw material, and these methods do not change the original form of baicalin in Astragalus, so Can be used instead of baicalin.
  • baicalin the solubility of baicalin compound is greatly improved, the absorption rate of baicalin is increased, the oral bioavailability is improved, and a water-soluble injection or powder injection can be conveniently prepared.
  • the compound can also exert synergistic pharmacological action of baicalin and magnesium ions to increase the drug effect.
  • baicalin powder Add to purified water and mix well to obtain a baicalin suspension.
  • the baicalin used in the present invention is a raw material for preparation and is a product widely sold on the market.
  • the ratio of baicalin to purified water is greater than 1:20, the concentration of baicalin suspension is too high, and the reaction is not completely caused to cause more impurities; if the ratio of baicalin to purified water is less than 1:100, The reaction rate of magnesium ion baicalin suspension is slow, and the reaction is not complete, resulting in more impurities; therefore, the ratio of baicalin to purified water is reasonable: 1:20-100, preferably 1:30-70; more preferably The ground is 1:32 ⁇ 36.
  • the magnesium compound is added to the baicalin suspension obtained in the step A, and uniformly mixed to obtain a baicalin suspension containing magnesium ions.
  • the magnesium compound used in the present invention is magnesium hydroxide, magnesium oxide, basic magnesium carbonate, magnesium acetate, magnesium sulfate, magnesium nitrate or magnesium chloride, which are currently widely marketed products.
  • the magnesium compound is magnesium hydroxide, magnesium oxide or basic magnesium carbonate.
  • the magnesium compound is magnesium hydroxide.
  • the molar ratio of baicalin to magnesium is less than 0.5:1, more magnesium ions remain in the reaction liquid, and are introduced into the product to form impurities; if the molar ratio of baicalin to magnesium is more than 3.0:1, The ratio of baicalin is too high, the reaction rate is too slow, it is difficult to filter and the impurities are more; therefore, the molar ratio of baicalin to magnesium is 0.5 to 3.0:1, preferably, the molar ratio of baicalin to magnesium is 0.8. ⁇ 2.6:1, more preferably, the molar ratio of baicalin to magnesium is from 1.8 to 2.2:1.
  • the magnesium ion-containing baicalin suspension obtained in the step B is allowed to react at a temperature of 20 to 70 ° C until the reaction system becomes clear and transparent, and filtered to obtain a filtrate.
  • the basic purpose of this step is to react the baicalin with the magnesium compound in the suspension of magnesium ion in step B to form the baicalin compound, that is, the Mg 2+ ion dissociated by the magnesium compound and the COO of baicalin in the reaction medium. - Ionic bonding to form a baicalin magnesium compound.
  • the temperature at which baicalin reacts with the magnesium compound is 20 to 70 ° C. If the reaction temperature is lower than 20 ° C, the reaction is too slow and the impurities are too much; if the reaction temperature is higher than At 70 ° C, baicalin is decomposed.
  • the reaction temperature is 40 to 65 ° C, and more preferably, the reaction temperature is 50 to 60 ° C.
  • the reaction system becomes clear and transparent to judge the progress of the reaction because the reaction belongs to a precipitation reaction in a solution, the reactant is a precipitate, the reaction product is dissolved in the reaction liquid, and the reaction system becomes clear and transparent, and the reaction can be judged. complete.
  • the basic purpose of the filtration is to remove residues which have not been completely reacted in the reaction system, or other impurities which do not participate in the reaction.
  • the filtering device used for filtration is various filtering devices commonly used in the field of pharmaceutical and chemical engineering, such as vacuum filters, plate and frame filter presses, and the like.
  • the filtrate obtained in the step C is dried to obtain the baicalin magnesium compound.
  • the drying method employed is heat drying, spray drying or freeze drying.
  • the above-described heating and evaporation is understood to mean that the water in the filtrate of the step C is heated and evaporated to obtain a baicalin compound having a water content of 1.0% by weight or less.
  • the method for determining the water content of the baicalin magnesium compound of the present invention is a drying method.
  • the spray drying is understood to be that the filtrate of the step C is dispersed into fine particles by a spray drying device, and the fine particles are brought into contact with hot air to remove the moisture in an instant, thereby obtaining magnesium baicaloside having a water content of 1.0% or less by weight.
  • the spray drying apparatus used in the present invention is currently marketed, for example, a product sold under the trade name QZR-5 by Linzhou Drying Machine Factory of Xishan City, Jiangsu province.
  • the freeze-drying is understood to be that the filtrate of step C is frozen to below 0 ° C by a freeze-drying apparatus, and the water is directly sublimed from the solid to water vapor by heating under high vacuum to remove water, and the water content is obtained by weight.
  • the baicalin magnesium compound is 1.0% or less.
  • the freeze-drying apparatus used in the present invention is a product currently on the market, such as a product sold by Beijing Sihuan Scientific Instrument Factory Co., Ltd. under the trade name LGJ-22D.
  • the invention further relates to a process for the preparation of another baicalin magnesium compound.
  • the macroporous adsorption resin is immersed in a 95% ethanol aqueous solution at a concentration for 20 to 28 hours, and then wet-packed, and then a 95% aqueous solution by volume is passed through the macroporous adsorption at a flow rate of 1.5 to 2.5 BV/h.
  • the resin column was no longer white turbid until the effluent and water were mixed at a volume ratio of 1:5, and then eluted with distilled water at a flow rate of 1.5 to 2.5 BV/h to be colorless.
  • the macroporous resin is understood to be a type of high molecular polymer which is concentrated and separated from the baicalin magnesium compound in the step C filtrate, and is also referred to as a polymer adsorbent.
  • the macroporous adsorption resin used in the present invention is selected from the group consisting of HPD-100, AB-8, D101 or YWD06B.
  • HPD-100 macroporous adsorption resin is a styrene-type non-polar copolymer, which is suitable for the extraction and separation of natural products such as saponins, flavonoids, and terpenoids.
  • the HPD-100 macroporous adsorption resin used in the present invention is a product sold by Zhengzhou Qinshi Technology Co., Ltd.
  • the AB-8 type macroporous adsorption resin is a styrene type weakly polar copolymer, which is suitable for extraction, separation or purification of weakly polar substances such as stevioside and alkaloid.
  • the AB-8 type macroporous adsorption resin used in the present invention is a product sold by Zhengzhou Qinshi Technology Co., Ltd.
  • D-101 type macroporous adsorption resin is a styrene type non-polar copolymer, which is a polymer adsorbent having a porous sponge structure, which is suitable for separation of saponins, flavonoids and alkaloids.
  • the D-101 type macroporous adsorption resin used in the present invention is a product sold by Zhengzhou Xidian Power Resin Sales Co., Ltd.
  • YWD06B macroporous adsorption resin is mainly used for the extraction of bilirubin, alkaloids and flavonoids, and also for the extraction of tea polyphenols, soy isoflavones, epimedium flavonoids and astragalus.
  • the YWD06B macroporous adsorption resin used in the present invention is a product sold by Zhengzhou Qinshi Technology Co., Ltd.
  • the main purpose of the macroporous adsorption resin pretreatment is to remove floating impurities and inert organic reagents in the macroporous adsorption resin.
  • the macroporous adsorption resin used in the present invention has a particle size of 10 to 80 mesh.
  • the ratio of the diameter of the macroporous adsorption resin column to the column height used in the present invention is 1:3 to 8.
  • the purpose of this extraction step was to extract baicalin from Astragalus using an aqueous ethanol solution.
  • the ratio of the xanthine to the aqueous ethanol solution is greater than 1:8, the concentration of the extract is too high, and the extraction is incomplete; if the ratio of the xanthine to the aqueous ethanol solution is less than 1:15, the volume of the extract is too large.
  • the adsorption in the macroporous adsorption resin is not good, and the impurities in the eluent are more. Therefore, a ratio of xanthine to aqueous ethanol solution of 1:8 to 15 is suitable.
  • the ratio of the xanthine to the aqueous ethanol solution is 1:9 to 12, and more preferably, the ratio of the xanthine to the aqueous ethanol solution is 1:9.6 to 10.4.
  • the concentration of the aqueous ethanol solution used for extracting the baicalin is less than 40%, the impurities in the extract are more; if the concentration of the aqueous ethanol solution used for extracting baicalin is higher than 60%, the jaundice is The magnesium glucoside extraction rate is lowered; therefore, it is reasonable to use a concentration of 40 to 60% of the aqueous ethanol solution used for extracting the baicalin, preferably 44 to 56%, more preferably 48 to 52%.
  • the temperature of extracting baicalin is lower than 55 °C, the extraction of magnesium baicalin may be incomplete; if the temperature of extracting baicalin is higher than 65 °C, decomposition of baicalin may occur; therefore, magnesium baicalin is extracted.
  • a temperature of 55 to 65 ° C is possible, preferably 57 to 63 ° C, more preferably 58 to 62 ° C.
  • the agitation speed is not critical and can be selected according to the actual situation, which is not difficult for those skilled in the art.
  • the extraction equipment used in this extraction step is an extraction concentrating unit, a hot reflux extraction unit, a static extraction unit or an electric and steam multi-function extraction unit, which are all products currently on the market, such as Jiangsu Changshu Traditional Chinese Medicine Pharmaceutical Machinery Factory.
  • the Astragalus membranaceus extract obtained in the step B is adsorbed by the macroporous adsorption resin column pretreated in the step A at a flow rate of 1.5 to 2.5 BV/h, and then washed with 4 to 6 macroporous adsorption resin bed volume water, and the washing water is discarded.
  • the baicalin is adsorbed on the macroporous adsorption resin and is not easily eluted; if the xanthine extract passes through the large
  • the flow rate of the pore-adsorbing resin column is higher than 2.5 BV/h, the baicalin is not well separated from the impurities, and the eluent has more impurities; therefore, the flow rate of the xanthine extract through the macroporous adsorption resin column is 1.5 to 2.5 BV.
  • /h is reasonable, preferably 1.7 to 2.3 BV/h, more preferably 1.9 to 2.1 BV/h.
  • the eluate containing the baicalin compound obtained in the step C is concentrated under reduced pressure at a temperature of 35 to 65 ° C until the volume of the concentrate is 1 to 15 times the weight of the astragalus medicinal material, and then dried to obtain a A crude extract of baicalin magnesium compound.
  • the eluate is concentrated under reduced pressure at a pressure of 0.01 to 0.1 MPa and a temperature of 35 to 65 °C.
  • the equipment used for concentration under reduced pressure is a product currently on the market, such as a product sold by Shanghai Shenshun Biotechnology Co., Ltd. under the trade name R2002 rotary evaporator.
  • the crude extract of the baicalin magnesium compound is purified in step D to obtain the baicalin magnesium compound.
  • the obtained baicalin compound is purified by recrystallization, octadecylsilane reverse phase column, and preparative liquid chromatography to achieve a content of the baicalin compound of 95% by weight or more.
  • recrystallization purification is carried out in an ethanol-water (1:9 to 9:1) solvent at 60 ° C in a water bath.
  • ODS octadecylsilane reverse phase column
  • the apparatus used for purification by preparative liquid chromatography was a preparative liquid phase sold by Shimadzu Corporation under the trade name CTO-10A, and the column was prepared to have a liquid phase condition of methanol-water (30:70).
  • baicalin magnesium compound was determined by high performance liquid chromatography using an instrument of Agilent 1260, Aglient 5HC C 18 (2), and methanol-water (40:60) in liquid phase.
  • the solubility of baicalin and baicalin in water at 37 ° C was 0.058 mg / mL and 129.1 mg / mL, respectively, which shows that the water content of baicalin compound is better, can be used completely For the preparation of water-soluble injections or freeze-dried powder injections.
  • Ultraviolet absorption spectroscopy was performed using the Agilent 8453 device.
  • Ultraviolet absorption spectrum analysis conditions automatically scan the maximum absorption wavelength from 200 to 760 nm.
  • the ultraviolet absorption spectrum was measured by a comparative method.
  • the equipment used for infrared absorption spectroscopy was a BRUKER Fourier transform infrared spectrometer, model TENSOR27.
  • the infrared absorption spectrum was measured by a conventional potassium bromide tableting method.
  • the carbonyl absorption peak on glucuronic acid is 1727 cm -1 , and the peak disappears after the compound is formed.
  • 1202cm -1 is the stretching vibration absorption peak of the 5-position CO.
  • baicalin reacts with Mg 2+ on the carboxyl group on the reaction glucuronic acid.
  • the infrared spectrum peaks of baicalin and baicalin are listed in Table 2 below. in.
  • Baicalin 13 C-NMR assignment Baicalin 13 C-NMR showed a total of 21 C signals, in which the 13 C-NMR of the baicalin compound was compared with the baicalin 13 C-NMR, the number of absorption peaks, chemical shift, The number of splits and the coupling constants of the peaks are very close, indicating that the intervention of Mg 2+ has little effect on the 13 C nuclear spin of baicalin, indicating that Mg 2+ is unlikely to bind to the C4 and C5 positions of baicalin.
  • 12.5 is hydrogen at the 5-position-OH
  • 12.7 is the active hydrogen at the carboxyl group on the glucuronic acid.
  • 12.5 still exists, and 12.7 disappears, and the number of other 1 H absorption peaks, chemical shift
  • the number of splits and the coupling constants of the peaks are very close, which further proves that Mg 2+ binds to baicalin on the glucuronate carboxyl group.
  • Mass spectrometry conditions ESI source; mobile phase: acetonitrile; mass spectrometry; scanning range: 100-2000.
  • the equipment used for the elemental analysis method was a P.E 2400 Elemental Analyzer Model 2400II.
  • the equipment used for atomic absorption analysis is SHIMADZU, AA-7000.
  • the atomic absorption analysis conditions are standard curve methods, and the method uses a conventional flame method.
  • the invention further relates to the use of said baicalin magnesium compound.
  • the same amount of baicalin compound may be used instead of xanthine or baicalin, or it may be prepared for treating liver injury, cerebral ischemia, diabetes, inflammation or Use in oncology drugs.
  • the preparation method of the baicalin magnesium in the invention does not need to add acid and alkali in the preparation process, does not destroy the original form of baicalin in the astragalus, does not use toxic and harmful organic reagents, is green and environmentally friendly, and has high product purity.
  • the method for extracting baicalin of the invention does not destroy the original form of baicalin in the astragalus, is green and environmentally friendly, has high purity of the product; the synthesis method is simple, the quality is controllable, and the original baicalin can be reduced in the original of astragalus There is a form of existence.
  • the solubility of baicalin is greatly improved, the absorption rate of baicalin is increased, the oral bioavailability is improved, and the baicalin can be substituted for the medicine, and the water-soluble injection or powder injection can be conveniently prepared.
  • Pharmacological experiments show that the magnesium baicalin has good pharmacological activity.
  • Figure 1 is a graph showing changes in body temperature of yeast-induced rats.
  • Figure 2 is a graph showing the effect of baicalin on yeast-induced rats.
  • the baicalin powder is added to the purified water, and uniformly mixed to obtain a baicalin suspension
  • the magnesium magnesium hydroxide compound is added to the baicalin suspension obtained in step A, and uniformly mixed to obtain a baicalin suspension containing magnesium ions;
  • the magnesium ion-containing baicalin suspension obtained in the step B is reacted at a temperature of 30 ° C until the reaction system becomes clear and transparent, and filtered using a Buchner funnel vacuum filtration apparatus to obtain a filtrate. ;
  • step C Let the filtrate obtained in step C be sold under the trade name of Wujiang Huafei Electric Heating Equipment Co., Ltd.
  • the apparatus of the HF881-2 drying oven was heated and evaporated to dryness at 60 ° C to obtain a baicalin compound having a water content of 1.0% by weight or less.
  • the baicalin powder is added to the purified water, and uniformly mixed to obtain a baicalin suspension
  • the magnesium magnesium oxide compound is added to the baicalin suspension obtained in step A, and uniformly mixed to obtain a baicalin suspension containing magnesium ions;
  • the magnesium ion-containing baicalin suspension obtained in the step B was reacted at a temperature of 70 ° C until the reaction system became clear and transparent, and filtered using a Buchner funnel vacuum filtration apparatus to obtain a filtrate. ;
  • the filtrate obtained in the step C was spray-dried using a spray drying equipment sold by the company of Linzhou Drying Machine Factory of Xishan City, Jiangsu province under the trade name QZR-5, to obtain a baicalin compound having a water content of 1.0% by weight or less.
  • baicalin powder is added to the purified water, and uniformly mixed to obtain a baicalin suspension
  • the basic magnesium magnesium carbonate compound is added to the baicalin suspension obtained in step A, and uniformly mixed to obtain a suspension containing baicalin containing magnesium ions;
  • the magnesium ion-containing baicalin suspension obtained in the step B is reacted at a temperature of 60 ° C until the reaction system becomes clear and transparent, and filtered using a Brookfield funnel vacuum filtration apparatus to obtain a filtrate. ;
  • the filtrate obtained in the step C is freeze-dried at a temperature of -40 ° C by a freeze drying apparatus of the trade name EYELA OSB-2100 by Beijing Wuzhou Oriental Technology Development Co., Ltd. to obtain baicalin having a water content of 1.0% or less by weight.
  • EYELA OSB-2100 by Beijing Wuzhou Oriental Technology Development Co., Ltd.
  • baicalin powder is added to the purified water, and uniformly mixed to obtain a baicalin suspension
  • the magnesium sulphate compound is added to the baicalin suspension obtained in step A, and uniformly mixed to obtain a baicalin suspension containing magnesium ions;
  • the magnesium ion-containing suspension obtained in the step B was reacted at a temperature of 70 ° C until the reaction system became clear and transparent, and filtered using a Buchner funnel vacuum filtration apparatus to obtain a filtrate;
  • the filtrate obtained in the step C was spray-dried using a spray drying equipment sold by the company of Linzhou Drying Machine Factory of Xishan City, Jiangsu province under the trade name QZR-5, to obtain a baicalin compound having a water content of 1.0% by weight or less.
  • the baicalin powder is added to the purified water, and uniformly mixed to obtain a baicalin suspension
  • the magnesium nitrate compound is added to the baicalin suspension obtained in step A, and uniformly mixed to obtain a baicalin suspension containing magnesium ions;
  • the magnesium ion-containing suspension obtained in the step B is reacted at a temperature of 60 ° C until the reaction system becomes clear and transparent, and filtered using a Brookfield funnel vacuum filtration apparatus to obtain a filtrate;
  • the filtrate obtained in the step C is freeze-dried at -40 ° C using a freeze-drying equipment sold by Beijing Wuzhou Oriental Technology Development Co., Ltd. under the trade name EYELA OSB-2100 to obtain jaundice having a water content of 1.0% or less by weight.
  • EYELA OSB-2100 a freeze-drying equipment sold by Beijing Wuzhou Oriental Technology Development Co., Ltd. under the trade name EYELA OSB-2100 to obtain jaundice having a water content of 1.0% or less by weight.
  • Magnesium glycinate compound Magnesium glycinate compound.
  • the baicalin powder is added to the purified water, and uniformly mixed to obtain a baicalin suspension
  • the magnesium magnesium compound is added to the baicalin suspension obtained in step A, and uniformly mixed to obtain a baicalin suspension containing magnesium ions;
  • the magnesium ion-containing suspension obtained in the step B was reacted at a temperature of 50 ° C until the reaction system became clear and transparent, and filtered using a Buchner funnel vacuum filtration apparatus to obtain a filtrate;
  • the filtrate obtained in the step C is freeze-dried at -40 ° C using a freeze-drying equipment sold by Beijing Wuzhou Oriental Technology Development Co., Ltd. under the trade name EYELA OSB-2100 to obtain jaundice having a water content of 1.0% or less by weight.
  • EYELA OSB-2100 a freeze-drying equipment sold by Beijing Wuzhou Oriental Technology Development Co., Ltd. under the trade name EYELA OSB-2100 to obtain jaundice having a water content of 1.0% or less by weight.
  • Magnesium glycinate compound Magnesium glycinate compound.
  • the macroporous adsorption resin sold by Zhengzhou Qinshi Technology Co., Ltd. under the trade name HPD-100 was immersed in a 95% ethanol aqueous solution at a concentration of 26 hours, and then packed in a wet manner, followed by a 95% ethanol aqueous solution by volume.
  • the resin column was adsorbed through the macroporous at a flow rate of 1.5 BV/h until the effluent and water were mixed at a volume ratio of 1:5, and then no white turbidity was observed, followed by elution with distilled water at a flow rate of 1.7 BV/h to a colorless state;
  • the xanthine was added to a 50% aqueous solution of the concentration by volume, and the mixture was extracted at a temperature of 60 ° C for 1.0 hour with stirring, and then separated to obtain a An extract and a residue; the residue is repeatedly extracted twice under the same conditions as above, and the residue is discarded; the extract is combined to obtain a xanthine extract;
  • Step B Let the Astragalus membranaceus extract obtained in Step B be adsorbed by the large pore adsorption resin column pretreated in Step A at a flow rate of 1.5 BV/h, and then washed with 4 large pore adsorption resin bed volume water, and the washing water is discarded; The volume concentration of the macroporous adsorption resin bed is eluted by 50% by volume aqueous solution of ethanol, and the eluate containing baicalin is collected;
  • the eluate containing the baicalin obtained in the step C is concentrated under reduced pressure at a pressure of 0.01 MPa and a temperature of 50 ° C using a device sold by Shanghai Shenshun Biotechnology Co., Ltd. under the trade name R2002 rotary evaporator until the concentrate is concentrated.
  • the volume is 1 times the weight of the Astragalus medicinal herbs in grams, and then evaporated to dryness at 60 ° C by the equipment of Wujiang Huafei Electric Heating Equipment Co., Ltd. under the trade name HF881-2 drying oven to obtain a crude extract of baicalin compound.
  • step D The crude extract of baicalin magnesium compound obtained in step D is recrystallized and purified in an 80% ethanol solvent at 60 ° C to obtain the baicalin magnesium compound.
  • the macroporous adsorption resin sold by Zhengzhou Qinshi Technology Co., Ltd. under the trade name AB-8 is immersed in a 95% ethanol aqueous solution at a concentration for 20 hours, and then packed in a wet manner, followed by a 95% ethanol aqueous solution by volume.
  • the resin column was adsorbed through the macroporous at a flow rate of 2.5 BV/h until the effluent and water were mixed at a volume ratio of 1:5, and then no white turbidity was observed, followed by elution with distilled water at a flow rate of 1.5 BV/h to a colorless state;
  • the xanthine was added to a 40% by weight aqueous solution of ethanol, and extracted at a temperature of 55 ° C for 1.5 hours with stirring, and then separated to obtain a Extract and a residue; the residue is repeatedly extracted 3 times under the same conditions as above, and the residue is discarded; the extract is combined to obtain a xanthine extract;
  • Step B Let the Astragalus membranaceus extract obtained in Step B be adsorbed at a flow rate of 2.5 BV/h through the macroporous adsorption resin column pretreated in Step A, and then washed with 5 large pore adsorption resin bed volume water, and the washing water is discarded;
  • the volume concentration of the macroporous adsorption resin bed is eluted by 45% by volume of an aqueous ethanol solution, and the eluate containing baicalin is collected;
  • the eluate containing the baicalin obtained in the step C is concentrated under reduced pressure at a pressure of 0.03 MPa and a temperature of 35 ° C using a device sold by Shanghai Shenshun Biotechnology Co., Ltd. under the trade name R2002 rotary evaporator until the concentrate
  • the volume is 15 times of the weight of the Astragalus medicinal herbs in grams, and then spray-dried by a spray drying device sold by Linzhou Drying Machine Factory of Jiangsu Xishan City under the trade name QZR-5 to obtain a crude extract of baicalin magnesium compound;
  • the crude extract of baicalin compound obtained in step D was purified by using an ODS column sold by YMC under the trade name GEL C18AAG12S50 in an amount of 10% to 40% methanol water as an eluent to obtain the baicalin compound.
  • the macroporous adsorption resin sold by Zhengzhou Xidian Power Resin Sales Co., Ltd. under the trade name D-101 was immersed in a 95% ethanol aqueous solution at a concentration of 28 hours, and then packed in a wet manner, followed by a concentration of 95% by volume.
  • the aqueous ethanol solution was passed through the macroporous adsorption resin column at a flow rate of 2.2 BV/h until the effluent and water were mixed at a volume ratio of 1:5, and then no white turbidity was observed, followed by elution with distilled water at a flow rate of 2.0 BV/h until colorless. ;
  • the xanthine was added to a 60% aqueous solution of the concentration by volume, and extracted at a temperature of 65 ° C for 1.0 hour under stirring, and then separated to obtain a An extract and a residue; the residue is repeatedly extracted twice under the same conditions as above, and the residue is discarded; the extract is combined to obtain a xanthine extract;
  • Step B Let the Astragalus membranaceus extract obtained in Step B be adsorbed by the large pore adsorption resin column pretreated in Step A at a flow rate of 2.0 BV/h, and then washed with 4 large pore adsorption resin bed volume water, and the washing water is discarded; The volume concentration of the macroporous adsorption resin bed is eluted by 55% by volume aqueous solution of ethanol, and the eluate containing baicalin is collected;
  • the eluate containing the baicalin obtained in the step C is concentrated under reduced pressure at a pressure of 0.1 MPa and a temperature of 65 ° C using a device sold by Shanghai Shenshun Biotechnology Co., Ltd. under the trade name R2002 rotary evaporator until the concentrate is
  • the volume is 5 times the weight of the Astragalus medicinal herbs in grams, and then freeze-dried at -40 ° C using a freeze-drying equipment sold by Beijing Sihuan Scientific Instrument Factory Co., Ltd. under the trade name LGJ-22D to obtain a magnesium baicalin.
  • the macroporous adsorption resin sold by Zhengzhou Qinshi Technology Co., Ltd. under the trade name YWD06B is soaked in a 95% ethanol aqueous solution at a concentration for 24 hours, then wet-packed, and then subjected to a concentration of 95% ethanol aqueous solution to 2.0.
  • the BV/h flow rate was passed through the macroporous adsorption resin column until the effluent and water were mixed at a volume ratio of 1:5, and then no longer appeared white turbid, and then eluted with distilled water at a flow rate of 2.5 BV/h to be colorless;
  • xanthine was added to a 45% by volume aqueous solution of ethanol, and extracted at a temperature of 65 ° C for 1.0 hour with stirring, and then separated to obtain a Extract and a residue; the residue is repeatedly extracted 3 times under the same conditions as above, and the residue is discarded; the extract is combined to obtain a xanthine extract;
  • Step B Let the Astragalus membranaceus extract obtained in Step B be adsorbed by the macroporous adsorption resin column pretreated in Step A at a flow rate of 2.0 BV/h, and then washed with 5 large pore adsorption resin bed volume water, and the washing water is discarded; The volume concentration of the macroporous adsorption resin bed is eluted by 50% by volume aqueous solution of ethanol, and the eluate containing baicalin is collected;
  • the eluate containing the baicalin obtained in the step C is concentrated under reduced pressure at a pressure of 0.07 MPa and a temperature of 45 ° C using a device sold by Shanghai Shenshun Biotechnology Co., Ltd. under the trade name R2002 rotary evaporator until the concentrate is
  • the volume is 10 times of the weight of the Astragalus medicinal herbs in grams, and then spray-dried by a spray drying device sold by the Linzhou Drying Machine Factory of Xishan City, Jiangsu province under the trade name QZR-5 to obtain a crude extract of baicalin magnesium compound;
  • the crude extract of the baicalin magnesium compound is used by the YMC company under the trade name.
  • the ODS column sold by GEL C18AAG12S50 was purified under the conditions of 10% to 40% methanol water as an eluent to obtain the baicalin compound.
  • mice Forty healthy mice were randomly divided into 4 groups: normal group, model group, baicalin group and baicalin group.
  • the baicalin group was administered at a dose of 10 mg/kg, and the baicalin group was administered at a dose equivalent to 10 mg/kg baicalin.
  • the model group was intragastrically administered with the same volume of normal saline for 5 days.
  • the model group and the administration group were intraperitoneally injected with D-galactose 0.5 g/kg, and the normal group was intraperitoneally injected with the same volume of physiological saline.
  • the activities of serum ALT, AST, SOD, GSH-Px and MDA content were measured. Analysis of variance was performed using SPSS-19.0 statistical software. The results are shown in Table 5.
  • mice Eighty healthy mice were randomly divided into 4 groups: model group, sham operation group, baicalin group, and baicalin group, 20 in each group.
  • mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital. After anesthesia, the left and right common carotid arteries were exposed and vascular clamps were used to clamp the bilateral arteries for 20 min each time. The vascular clamp was loosened, sutured, and placed.
  • sham operation group the same method was used to expose the left and right common carotid arteries, but not clipped, the wound was sutured and placed in the cage for feeding;
  • baicalin group operation with the model group, On the 2nd day after the operation, 12.5 mg/kg baicalin was intraperitoneally injected for 12 days; the baicalin group was treated with the model group, and the baicalin was intraperitoneally injected on the 2nd day after surgery (equivalent to baicalin 12.5 mg/kg). ), continuous injection for 12 days, the sham operation group and the model group were given an equal volume of physiological saline.
  • the circular pool for testing has a diameter of 120 cm and a height of 60 cm.
  • the cylindrical platform has a diameter of 10 cm and a height of 40 cm.
  • the inside of the pool is black.
  • a camera connected to the display is placed above, and the movement trajectory of the mouse is recorded.
  • the water depth was 12 cm and the water temperature was (25 ⁇ 2) °C. All the space marks around the maze remained unchanged throughout the experiment.
  • Water maze memory test experiment One day before the start of the formal experiment, each mouse was familiar with the water environment for 2 min in the maze, and the formal experiment was started on the second day. The mice were randomly and from the four different quadrants (North, West, South, East, for a period of time) into the water, and the time when the mice entered the water to climb the platform was recorded, which is the escape latency. If the platform is not found within 120 seconds, the incubation period is recorded as 120 seconds, and it is guided to find the platform and rest for 60 seconds. Continuous training for 15 days, 2 times / day. Each group of animals was subjected to a water maze test in turn.
  • the first day of the path included 2 blind ends, 3 blind ends on the 2nd day, and 4 blind ends on the 3rd, 4th, and 5th days.
  • the mice were placed in the water at the starting point facing the pool wall, and the number of times the mouse was placed in the water to reach the end platform (latency period) and the number of blind spots was recorded. If the mouse does not reach the platform within 60 seconds, the mouse can be rushed to the platform, and the mouse is allowed to stay on the platform for 20 seconds, and each training is performed once a day in the afternoon and the experiment for 5 days.
  • mice were randomly divided into 3 groups. They were baicalin group, baicalin group and control group. Each group was divided into 6 groups according to time points 1, 3, 5, 7, 11, 15h, with 8 groups in each group.
  • the baicalin group dose was 100 mg ⁇ kg -1
  • the baicalin dose was equivalent to 100 mg ⁇ kg -1 baicalin.
  • the rats in the control group were given the same volume of physiological saline. Each group was given 100 ⁇ l of xylene to the left ear of the mice 40 minutes before the arrival of each time point. When the time was reached, the animals were sacrificed and the ears of the ears of the mice were taken with a puncher to the weight of the right ear and the left ear. The difference (swelling degree) is an indicator to observe the anti-inflammatory effect of the drug, and the results are shown in Table 7.
  • Human hepatoma HepG2 cells were cultured in DMEM medium containing 10% calf serum, penicillin (100 ⁇ mol ⁇ L -1 ) and streptomycin (1 mg ⁇ mL -1 ) at 37 ° C in 5% CO 2 incubator. .
  • HepG2 cells in logarithmic growth phase were diluted with 10% fetal bovine serum RPMI-1640 medium to a concentration of 1 ⁇ 10 4 cells/mL, and seeded in 96-well plates at 100 ⁇ L per well.
  • 37 ° C, relative humidity 90% and 5% CO 2 incubator culture cultured for 24h, discarded the culture solution, respectively added baicalin and baicalin magnesium (equivalent to baicalin concentration (5,10,20 ⁇ g/mL))
  • baicalin and baicalin magnesium equivalent to baicalin concentration (5,10,20 ⁇ g/mL)
  • Parallel operation was performed in parallel with the test well without adding cells to the blank control wells of the culture medium, and only the negative control group without cells was added, and the cells were further cultured for 48 hours to collect the cells. After washing twice with PBS solution, it was detected by flow cytometry FACS, and parameter acquisition and data analysis were performed. The results are shown in Table 8.
  • mice weighing 20-22 g, were randomly divided into 4 groups, namely baicalin 0.067 g/kg group, baicalin magnesium (corresponding to baicalin 0.067 g/kg), blank control group (administered saline).
  • Aspirin-positive control group (0.15 g/kg), 10 in each group.
  • the drug was administered once a day for 7 consecutive days. 30 min after the last administration, 0.2 mL/10 g of 0.6% acetic acid physiological saline solution was intraperitoneally injected to observe the number of writhing reactions in the mice within 15 min, and the analgesic rate was calculated.
  • the analgesic rate [(the average writhing frequency of the blank control group) - the average number of writhings in the experimental group) / the average number of writhing in the blank control group] ⁇ 100%.
  • One-way analysis of variance was performed on the data using SPSS19.0 statistical software. P ⁇ 0.05 was considered statistically significant. The results are shown in Table 9.
  • the animal feeding environment is a regular day and night cycle, room temperature (22 ⁇ 2) ° C, relative humidity (50 ⁇ 2)%.
  • the rats were placed in the experimental environment and simulated to adapt to the experimental conditions. Rats with basal body temperature between 37.5 and 39.0 °C were selected, and the experiment was started on the fourth day. Fasting for 12 hours before the experiment, the body temperature was measured once every 1 hour after the start of the experiment, and the average value was taken as the basal body temperature. The difference between the highest and lowest values in the sub-temperature was greater than 0.5 °C.
  • the baicalin or baicalin magnesium was administered intragastrically, and then immediately injected with a 20% dry yeast physiological saline suspension (except for the control group).
  • the administration methods were as follows: control group 2 ml/kg saline + subcutaneous injection of physiological saline 10 ml/kg, model group 2 ml/kg saline + subcutaneous injection 2 g/kg, baicalin group 80 mg/kg + subcutaneous injection Yeast 2g/kg, baicalin magnesium group (equivalent to 80mg/kg baicalin) + subcutaneous injection of yeast 2g/kg, after administration, body temperature was measured every 2h, and the body temperature change at different time after injection of yeast was observed ( ⁇ T , ° C), continuous measurement 14h, the results are shown in Figure 1 and Figure 2.
  • the model group was also significantly lower (P ⁇ 0.05). Meanwhile, compared with baicalin, the ⁇ T was 0.2 and 0.5 °C at 6h and 8h. The results showed that baicalin inhibited or interfered with yeast-induced fever. Magnesium glucoside is better.
  • the administration was started after successful modeling, and the drug was administered intragastrically every morning.
  • the baicalin group dose was 80 mg/kg and the baicalin group (corresponding to baicalin 80 mg/kg).
  • the diabetic model and the high-fat diet control group were intragastrically administered with the same volume of solvent once a day for 6 weeks. Blood glucose was measured after modeling and at 3 and 6 weeks after administration.
  • One-way analysis of variance was performed on the data using SPSS19.0 statistical software. P ⁇ 0.05 was considered statistically significant. The results are shown in Table 10.
  • the results in Table 10 indicate that at the third week of the experiment, the blood glucose of the diabetic model group was increased, and the blood glucose of the baicalin group and the baicalin group was decreased, respectively.
  • the blood glucose of the diabetic model group was still increasing, while the blood glucose of the baicalin group and the baicalin group was further decreased, and the blood glucose levels of the baicalin and baicalin group were significantly lower than those of the diabetic model group (P ⁇ 0.05).
  • the blood glucose level of the baicalin group was significantly stronger than that of the baicalin group (P ⁇ 0.05).

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Abstract

一种黄芩苷镁化合物，还涉及黄芩苷镁化合物制备方法和提取方法。所述制备方法包括制备黄芩苷混悬液、制备含镁离子混悬液、反应与干燥等步骤；所述提取方法包括大孔吸附树脂预处理、提取、吸附与洗脱、浓缩与干燥、纯化等步骤。制备黄芩苷镁还原了黄芩苷在黄芩中的存在方式，可以代替黄芩苷入药。该黄芩苷镁化合物相比黄芩苷溶解度大大提高，口服吸收速度比黄芩苷快，生物利用度比黄芩苷高，还可以非常方便制成水溶性注射剂或粉针剂。药理实验表明，所述的黄芩苷镁的药理活性优于黄芩苷。

Description

一种黄芩苷镁化合物及其制备方法与它的用途
本发明请求申请日为2016年02月01日、申请号为CN 2016100689263的中国发明专利申请为优先权。
【技术领域】
本发明属于医药技术领域。更具体地，本发明涉及一种黄芩苷镁化合物，还涉及所述黄芩苷镁化合物的提取、制备方法，涉及所述黄芩苷镁化合物的用途。
【背景技术】
黄芩首载于《神农本草经》，为唇形科植物黄芩Scutellaria baicalensis Georgi的干燥根，具有清热燥湿、泻火解毒、止血安胎的功能。黄芩苷是一种黄芩含有的黄酮类化合物，分子式为C21H18O11，在黄芩中含量为以重量计9％以上，甚至高达以重量计20％，是黄芩的主要有效成分之一。
现代药理学研究表明，黄芩苷具有清除氧自由基、抗心律失常、扩张心脑血管、降压镇静、保肝利胆、抗病原体作用，对前列腺癌、宫颈癌、肺癌、恶性淋巴瘤、骨髓瘤、肝癌等肿瘤也有抑制作用，在临床上已有黄芩苷片、黄芩苷胶囊等制剂，主要用于治疗急慢性肝炎、心血管疾病等。
镁是一种参与生物体正常生命活动及新陈代谢过程的必不可少的元素。镁影响细胞的多种生物功能，影响钾离子和钙离子转运，调控信号传递，参与能量代谢、蛋白质和核酸合成；催化酶的激活和抑制及对细胞周期、细胞增殖及细胞分化的调控；镁还参与维持基因组的稳定性，并且还与机体氧化应激和肿瘤发生有关。
黄芩苷可溶于氢氧化钠、碳酸钠、碳酸氢钠等碱性溶液中，但在碱液中不稳定，会渐变暗棕色，微溶于热冰醋酸，难溶于甲醇、乙醇、丙 酮，不溶于水、苯、乙醚、氯仿。由于黄芩苷几乎不溶于水，口服生物利用度低，极大地限制了其在临床上的广泛应用。
近年，利用新制剂技术提高黄芩苷溶解度和生物利用度的相关报道较多，主要涉及将黄芩苷制备成金属配合物、β环糊精包合物、固体分散体、磷脂复合物、微粒、纳米粒、脂质体纳米乳、微乳、明胶微球等。
目前，《中国药典》2015年版一部在黄芩提取物项下说明，采用水提酸沉提取，经碱溶酸沉纯化获得黄芩苷，目前市售黄芩苷都是采用这种方法制备得到的。黄芩苷在黄芩中的原本存在形式是镁盐形式，这种形式的水溶性很好，可以通过水溶解提取，但加酸沉淀破坏了其原有结构，使得黄芩苷的水溶性变差，吸收速度慢，生物利用度低，而且黄芩苷药效也有所降低。
本发明找到一种在不改变黄芩中黄芩苷的原有存在方式基础上从黄芩中提取出黄芩苷镁化合物的方法，同时也找到了以市售黄芩苷为原料合成黄芩苷镁化合物的方法，还原了黄芩苷在黄芩中的存在方式。该黄芩苷镁化合物可以代替黄芩苷入药，相比黄芩苷溶解度大大提高，能使黄芩苷的吸收速率变快，口服生物利用度提高，还可以非常方便制成水溶性注射剂或粉针剂。同时，该化合物还可以发挥黄芩苷与镁离子的协同药理作用，增加药效。
本发明人在总结现有技术的基础之上，通过大量实验研究分析，终于完成了本发明。
【发明内容】
[要解决的技术问题]
本发明的目的是提供一种黄芩苷镁化合物。
本发明的另一个目的是提供所述黄芩苷镁化合物的制备方法。
本发明的另一个目的是提供所述黄芩苷镁化合物的用途。
[技术方案]
本发明是通过下述技术方案实现的。
本发明涉及一种黄芩苷镁化合物。
所述的黄芩苷镁化合物是具有下述化学结构式(I)的化合物：
Figure PCTCN2017071616-appb-000001
本发明还涉及一种黄芩苷镁化合物制备方法。
该制备方法的步骤如下:
A、制备黄芩苷混悬液
按照以克计黄芩苷与以毫升计纯化水的比为1:20～100，把黄芩苷粉加到纯化水中，混合均匀，得到一种黄芩苷混悬液。
B、制备含镁离子的黄芩苷混悬液
按照黄芩苷与镁的摩尔比0.5～3.0:1，往步骤A得到的黄芩苷混悬液中添加镁化合物，混合均匀，得到一种含镁离子的黄芩苷混悬液；
C、反应
在搅拌的条件下，让步骤B得到的含镁离子的黄芩苷混悬液在温度20～70℃下进行反应，直到其反应体系变得澄清透明，过滤，得到滤液；
D、干燥
将步骤C得到的滤液进行干燥，得到所述的黄芩苷镁化合物。
根据本发明的一种优选实施方式，所述的镁化合物选自氢氧化镁、氧化镁、碱式碳酸镁、乙酸镁、硫酸镁、硝酸镁或氯化镁。
本发明还涉及另一种黄芩苷镁化合物的制备方法。
该制备方法的步骤如下：
A、大孔吸附树脂预处理
将大孔吸附树脂在浓度以体积计95％乙醇水溶液中浸泡20～28小时，然后湿法装柱，接着让浓度以体积计95％乙醇水溶液以1.5～2.5BV/h 流速通过该大孔吸附树脂柱，直到流出液与水按照体积比1:5混合后不再呈现白色浑浊状，接着用蒸馏水以1.5～2.5BV/h流速洗脱至无色；
B、提取
按照以克计黄芩与以ml计乙醇水溶液的比为1:8～15，把黄芩加到浓度以体积计40～60％乙醇水溶液中，在温度55～65℃与搅拌的条件下提取0.8～1.2小时，然后分离，得到一种提取液与一种残留物；所述的残留物在与上述相同条件下重复提取2～3次，提余残留物弃去；提取液合并，得到一种黄芩提取液；
C、吸附与洗脱
让步骤B得到的黄芩提取液以1.5～2.5BV/h流速通过步骤A预处理的大孔吸附树脂柱进行吸附，然后使用4～6个大孔吸附树脂床体积水洗涤，洗涤水弃去；再用4～6个大孔吸附树脂床体积浓度为以体积计45～55％的乙醇水溶液洗脱，收集含有黄芩苷的洗脱液；
D、浓缩与干燥
将步骤C得到的含有黄芩苷的洗脱液，在温度35～65℃条件下减压浓缩，直至浓缩液的体积为以克计黄芩药材重量的1～15倍，再经干燥得到一种黄芩苷镁化合物粗提物；
E、纯化
对步骤D得到黄芩苷镁化合物粗提物进行纯化，得到所述的黄芩苷镁化合物。
根据本发明的另一种优选实施方式，所述的大孔吸附树脂选自HPD-100、AB-8、D101或YWD06B。
根据本发明的另一种优选实施方式，所述大孔吸附树脂的粒度10～80目。
根据本发明的另一种优选实施方式，所述大孔吸附树脂柱的直径与柱高之比是1：3～8。
根据本发明的另一种优选实施方式，所述的干燥是加热蒸干、喷雾干燥或冷冻干燥。
根据本发明的另一种优选实施方式，得到的黄芩苷镁化合物采用重结晶、十八烷基硅烷反相柱、制备液相色谱法进行纯化，以达到黄芩苷镁化合物含量为以重量计95％以上。
本发明还涉及所述的黄芩苷镁化合物的用途。所述的黄芩苷镁化合物以等剂量代替在含黄芩或黄芩苷的中药单方或复方制剂中的黄芩或黄芩苷，或者它在制备用于治疗肝损伤、脑缺血、糖尿病、炎症或肿瘤药物中的用途。还可以非常方便制成水溶性注射剂或粉针剂。
下面将更详细地描述本发明。
本发明涉及一种黄芩苷镁化合物。
所述的黄芩苷镁化合物是具有下述化学结构式(I)的化合物：
Figure PCTCN2017071616-appb-000002
本发明还涉及一种黄芩苷镁化合物制备方法。
本发明提出一种从黄芩中提取黄芩苷镁化合物的方法，同时还提出一种以市售黄芩苷为原料合成黄芩苷镁化合物的方法，这些方法没有改变黄芩苷在黄芩中原有存在形态，因此，可以代替黄芩苷入药。与黄芩苷相比，黄芩苷镁化合物溶解度大大提高，能使黄芩苷的吸收速率变快，口服生物利用度提高，能非常方便地制成水溶性注射剂或粉针剂。同时，该化合物还可以发挥黄芩苷与镁离子协同药理作用，增加药效。
该黄芩苷镁化合物制备方法的步骤如下:
A、制备黄芩苷混悬液
按照以克计黄芩苷与以毫升计纯化水的比为1:20～100，把黄芩苷粉 加到纯化水中，混合均匀，得到一种黄芩苷混悬液。
本发明使用的黄芩苷是制剂原料药，是目前市场上广泛销售的产品。
根据本发明，如果黄芩苷与纯化水的比大于1:20，则黄芩苷混悬液浓度太高，反应不完全致使杂质较多；如果黄芩苷与纯化水的比小于1:100，则含镁离子黄芩苷混悬液反应速度较慢，反应不完全致使杂质较多；因此，黄芩苷与纯化水的比为1:20～100是合理的，优选地是1:30～70；更优选地是1:32～36。
B、制备含镁离子的黄芩苷混悬液
按照黄芩苷与镁的摩尔比0.5～3.0:1，往步骤A得到的黄芩苷混悬液中添加镁化合物，混合均匀，得到一种含镁离子的黄芩苷混悬液。
本发明使用的镁化合物是氢氧化镁、氧化镁、碱式碳酸镁、乙酸镁、硫酸镁、硝酸镁或氯化镁，它们都是目前市场上广泛销售的产品。
优选地，所述的镁化合物是氢氧化镁、氧化镁或碱式碳酸镁。
更优选地，所述的镁化合物是氢氧化镁。
根据本发明，如果黄芩苷与镁的摩尔比小于0.5:1，则会反应液中剩余的镁离子较多，引入到产物中形成杂质；如果黄芩苷与镁的摩尔比大于3.0:1，则会黄芩苷比例过高，反应速度太慢，不易过滤且杂质较多；因此，黄芩苷与镁的摩尔比为0.5～3.0:1是恰当的，优选地，黄芩苷与镁的摩尔比为0.8～2.6:1，更优选地，黄芩苷与镁的摩尔比为1.8～2.2:1。
C、反应
在搅拌的条件下，让步骤B得到的含镁离子的黄芩苷混悬液在温度20～70℃下进行反应，直到其反应体系变得澄清透明，过滤，得到滤液。
这个步骤的基本目的在于让步骤B含镁离子的黄芩苷混悬液中的黄芩苷与镁化合物反应生成黄芩苷镁化合物，即在反应介质中镁化合物离解的Mg2+离子与黄芩苷的COO-离子键合生成黄芩苷镁化合物。
根据本发明，黄芩苷与镁化合物进行反应的温度为20～70℃，如果其反应温度低于20℃，则会反应太慢，杂质太多；如果其反应温度高于 70℃，则会黄芩苷发生分解。优选地，该反应温度为40～65℃，更优选地，该反应温度为50～60℃。
在本发明中，以其反应体系变得澄清透明来判断反应进程是因为该反应属于溶液中沉淀反应，反应物为沉淀物，反应产物溶于反应液中，反应体系变澄清透明基本可以判定反应完全。
在这个步骤中，过滤的基本目的是除去其反应体系中尚未反应完全的残留物，或者是不参与反应的其它杂质。
在这个步骤中，过滤使用的过滤设备是目前医药化工技术领域里通常使用的各种过滤设备，例如真空过滤机、板框压滤机等。
D、干燥
将步骤C得到的滤液进行干燥，得到所述的黄芩苷镁化合物。
在本发明中，采用的干燥方法是加热蒸干、喷雾干燥或冷冻干燥。
在本发明中，所述的加热蒸干应该理解是指将步骤C的滤液中的水分加热蒸发除去，得到水含量为以重量计1.0％以下的黄芩苷镁化合物。
本发明黄芩苷镁化合物中水含量测定方法是烘干法。
所述的喷雾干燥应该理解是采用喷雾干燥设备将步骤C的滤液分散成微粒，并且让其微粒与热空气接触，在瞬间除去其水分，得到水含量为以重量计1.0％以下的黄芩苷镁化合物。本发明使用的喷雾干燥设备是目前市场上销售的产品，例如由江苏锡山市林州干燥机厂公司以商品名QZR-5型销售的产品。
所述的冷冻干燥应该理解是采用冷冻干燥设备将将步骤C的滤液冷冻至0℃以下，并在高真空下通过加热使水分直接从固态升华为水汽，除去其水分，得到水含量为以重量计1.0％以下的黄芩苷镁化合物。本发明使用的冷冻干燥设备是目前市场上销售的产品，例如由北京四环科学仪器厂有限公司以商品名LGJ-22D型销售的产品。
本发明还涉及另一种黄芩苷镁化合物的制备方法。
该制备方法的步骤如下：
A、大孔吸附树脂预处理
将大孔吸附树脂在浓度以体积计95％乙醇水溶液中浸泡20～28小时，然后湿法装柱，接着让浓度以体积计95％乙醇水溶液以1.5～2.5BV/h流速通过该大孔吸附树脂柱，直到流出液与水按照体积比1:5混合后不再呈现白色浑浊状，接着用蒸馏水以1.5～2.5BV/h流速洗脱至无色。
在本发明中，所述的大孔树脂应该理解是一类对步骤C滤液中黄芩苷镁化合物具有浓缩、分离作用的高分子聚合物，也称之聚合物吸附剂。
本发明使用的大孔吸附树脂选自HPD-100、AB-8、D101或YWD06B。
HPD-100型大孔吸附树脂是苯乙烯型非极性共聚体，它适用于皂甙类、黄酮类、萜类等天然产物及植物的提取分离。本发明使用的HPD-100型大孔吸附树脂是郑州勤实科技有限公司销售的产品。
AB-8型大孔吸附树脂是苯乙烯型弱极性共聚体，它适宜于如甜菊苷、生物碱等具有弱极性物质的提取、分离或纯化。本发明使用的AB-8型大孔吸附树脂是郑州勤实科技有限公司销售的产品。
D-101型大孔吸附树脂是苯乙烯型非极性共聚体，是一种具有多孔海绵状结构的聚合物吸附剂，它适用于皂甙类、黄酮类、生物碱类分离。本发明使用的D-101型大孔吸附树脂是郑州西电电力树脂销售有限公司销售的产品。
YWD06B型大孔吸附树脂是主要用于胆红素，生物碱及黄酮类提取，也用于茶多酚，大豆异黄酮，淫羊藿黄酮及黄芪甙提取。本发明使用的YWD06B型大孔吸附树脂是郑州勤实科技有限公司销售的产品。
大孔吸附树脂预处理的主要目的是除去大孔吸附树脂中的漂浮杂质以及惰性有机试剂。
本发明使用的大孔吸附树脂的粒度10～80目。
本发明使用的大孔吸附树脂柱的直径与柱高之比是1:3～8。
B、提取
按照以克计黄芩与以ml计乙醇水溶液的比为1:8～15，把黄芩加到浓度以体积计40～60％乙醇水溶液中，在温度55～65℃与搅拌的条件下 提取0.8～1.2小时，然后分离，得到一种提取液与一种残留物；所述的残留物在与上述相同条件下重复提取2～3次，提余残留物弃去；提取液合并，得到一种黄芩提取液。
这个提取步骤的目的使用乙醇水溶液从黄芩提取黄芩苷。在本发明中，如果黄芩与乙醇水溶液的比大于1:8时，则会提取液浓度太高，提取不完全；如果黄芩与乙醇水溶液的比小于1:15时，则会提取液体积太大，在大孔吸附树脂中吸附不好，洗脱液杂质较多。因此，黄芩与乙醇水溶液的比为1:8～15是合适的。优选地，黄芩与乙醇水溶液的比是1:9～12，更优选地，黄芩与乙醇水溶液的比是1:9.6～10.4。
在本发明中，如果提取黄芩苷镁所使用乙醇水溶液的浓度低于40％时，则会提取液中杂质较多；如果提取黄芩苷所使用乙醇水溶液的浓度高于60％时，则会黄芩苷镁提取率下降；因此，提取黄芩苷镁所使用乙醇水溶液的浓度为40～60％是合理的，优选地是44～56％，更优选地是48～52％。
如果提取黄芩苷镁的温度低于55℃时，则会致使黄芩苷镁提取不完全；如果提取黄芩苷镁的温度高于65℃时，则会发生黄芩苷镁分解；因此，提取黄芩苷镁的温度为55～65℃是可行的，优选地是57～63℃，更优选地是58～62℃。
在这个步骤中，搅拌速度不是非常关键，可以根据实际进行选择，这对于本技术领域的技术人员而言是不存在任何困难的。
在这个提取步骤中使用的提取设备是提取浓缩机组、热回流提取机组、静态提取机组或电、汽多功能提取机组，它们都是目前市场上销售的产品，例如由江苏常熟中药制药机械总厂以商品名nR.ZN20A型销售的产品。
C、吸附与洗脱
让步骤B得到的黄芩提取液以1.5～2.5BV/h流速通过步骤A预处理的大孔吸附树脂柱进行吸附，然后使用4～6个大孔吸附树脂床体积水洗涤，洗涤水弃去。
在这个步骤中，如果所述黄芩提取液通过大孔吸附树脂柱的流速低于1.5BV/h，则会黄芩苷吸附于大孔吸附树脂上，不易洗脱；如果所述黄芩提取液通过大孔吸附树脂柱的流速高于2.5BV/h，则会黄芩苷与杂质分离不好，洗脱液杂质较多；因此，所述黄芩提取液通过大孔吸附树脂柱的流速为1.5～2.5BV/h是合理的，优选地是1.7～2.3BV/h，更优选地是1.9～2.1BV/h。
这个步骤使用水洗涤的基本目的在于除去杂质。如果洗涤水的量超过所述范围是不合理的，因为容易造成黄芩苷镁化合物的损失。
再用4～6个大孔吸附树脂床体积浓度为以体积计45～55％的乙醇水溶液洗脱，收集含有黄芩苷的洗脱液；
D、浓缩与干燥
将步骤C得到的含有黄芩苷镁化合物的洗脱液，在温度35～65℃条件下减压浓缩，直至浓缩液的体积为以克计黄芩药材重量的1～15倍，再经干燥得到一种黄芩苷镁化合物粗提物。
所述的洗脱液是在压力0.01～0.1MPa与温度35～65℃的条件下进行减压浓缩的。减压浓缩使用的设备是目前市场上销售的产品，例如由上海申顺生物科技有限公司以商品名R2002型旋转蒸发器销售的产品。
在这个步骤中，有关干燥的情况如前所述，在此不再赘述。
E、纯化
对步骤D得到黄芩苷镁化合物粗提物进行纯化，得到所述的黄芩苷镁化合物。
根据本发明，得到的黄芩苷镁化合物采用重结晶、十八烷基硅烷反相柱、制备液相色谱法进行纯化，以达到黄芩苷镁化合物含量为以重量计95％以上。
在本发明中，重结晶纯化是在乙醇-水(1：9～9:1)溶剂中在60℃水浴条件下进行的。
十八烷基硅烷反相柱(ODS)是一种常用的反相色谱柱，也叫C18柱；由于它是长链烷基键合相，有较高的碳含量和更好的疏水性，对各 种类型的生物大分子有更强的适应能力，因此在生物化学分析工作中应用的最为广泛。
使用ODS柱纯化是在使用由YMC公司以商品名GEL C18AAG12S50销售的ODS柱在10％～40％甲醇水作为洗脱液条件下进行的。
使用制备液相色谱法纯化所用仪器为岛津公司以商品名CTO-10A销售的制备液相，制备色谱柱为，液相条件为甲醇-水(30：70)。
黄芩苷镁化合物含量是根据高效液相色谱法测定的，所用仪器为Agilent 1260，色谱柱为Aglient 5HC C18(2)，液相条件为甲醇-水(40：60)。
根据平衡溶解度测定方法测定了黄芩苷与黄芩苷镁在温度37℃下在水中的溶解度分别是0.058mg/mL与129.1mg/mL，由此可见黄芩苷镁化合物的水溶性较好，完全可以用于制备水溶性注射液或冻干粉针剂。
下面将详细地描述采用现有化学结构分析方法对两种方法制备的黄芩苷镁化合物进行结构分析。
一、紫外吸收光谱分析
紫外吸收光谱分析使用的设备Agilent 8453。
紫外吸收光谱分析条件200～760nm下仪器自动扫描最大吸收波长。
采用对照品比较法测定紫外吸收光谱。
将2毫克黄芩苷与2毫克本发明制备的黄芩苷镁分别溶于10ml甲醇中，然后测定最大吸收波长，其结果列于表1中。
表1：黄芩苷与黄芩苷镁的最大吸收波长
Figure PCTCN2017071616-appb-000003
从表1可以看出，黄芩苷与黄芩苷镁化合物的紫外最大吸收波长(λmax)相同，它们都在216nm、276nm与317nm处有最大吸收；这些紫 外吸收光谱结果表明，镁离子的结合对黄芩苷的离域共振结构影响不大。而如果Mg2+结合于黄芩苷的4位碳基和5位酚羟基上，则整个分子中电子的离域程度将增大，致使电子跃迁时需要的能量降低，进而会使吸收峰发生红移。因此该结果提示Mg2+不大可能与黄芩苷的C4，C5配位。
二、红外吸收光谱
红外吸收光谱分析使用的设备为BRUKER傅立叶变换红外光谱仪，型号为TENSOR27。
采用常规溴化钾压片法测定红外吸收光谱。
通常醇、酚的游离-OH键吸收峰位于3650～3580cm-1，而缔合-OH键吸收峰位于3400～3200cm-1，而羧酸-OH键吸收峰位于3540～3350cm-1，黄芩苷在3390cm-1处有一个宽强峰，可归属于该分子中-OH的伸缩振动峰，黄芩苷镁位于3390cm-1的峰强度明显减弱，说明分子内的氢键被破坏。
通常C＝O吸收峰位于1850～1600cm-1，在黄芩苷红外光谱中，葡萄糖醛酸上的羰基吸收峰位于1727cm-1，在形成化合物后该峰消失。1660cm-1为4位C＝O吸收峰，1202cm-1为5位C-O的伸缩振动吸收峰，在形成化合物后无明显变化，位于1600cm-1～1400cm-1的苯环骨架吸收峰也无变化，这说明反应产物对苯环共轭体系无影响。通过对黄芩苷和黄芩苷镁红外光谱主要特征峰的比较分析，说明黄芩苷与Mg2+在反应葡萄糖醛酸上的羧基发生反应，黄芩苷和黄芩苷镁红外光谱峰归属列于下表2中。
表2：黄芩苷和黄芩苷镁红外光谱峰指定
Figure PCTCN2017071616-appb-000004
三、NMR氢谱碳谱
NMR分析使用的设备VARIAN INOVA 600。
NMR分析条件溶剂：氘代DMSO；测定氢谱碳谱。
黄芩苷、合成黄芩苷镁与充黄芩中提取黄芩苷镁的1H和13C-NMR数据(DMSO)列于表3中。
表3：黄芩苷与黄芩苷镁的NMR数据
Figure PCTCN2017071616-appb-000005
黄芩苷13C-NMR归属：黄芩苷13C-NMR共显示21个C信号，其中黄芩苷镁化合物的13C-NMR与黄芩苷13C-NMR相比，吸收峰的组数、化学位移、峰的分裂个数及偶合常数均十分接近，表明Mg2+的介入对黄芩苷的13C原子核自旋影响不大，表明Mg2+不大可能结合在黄芩苷的C4，C5位上。
在1H-NMR谱中12.5为5位-OH上氢，12.7为葡萄糖醛酸上羧基活泼氢，在形成镁化合物后12.5仍存在，而12.7消失，其余1H吸收峰的组数、化学位移、峰的分裂个数及偶合常数均十分接近，进一步证明了Mg2+与黄芩苷在葡萄糖醛酸上羧基结合。
四、质谱分析
质谱分析使用的设备Agilent 6310。
质谱分析条件ESI源；流动相：乙腈；2级质谱；扫描范围：100～2000。
质谱分析结果如下：
分子离子峰M/Z：914.9
M/Z：270.7，表明有较完整的黄芩苷苷元(C15H10O5)碎片；
M/Z：446.7，表明有黄芩苷(C21H18O11)碎片；
M/Z：468.6，表明有黄芩苷镁(C21H17O11Mg)碎片。
五、元素分析和原子吸收分析
元素分析方法使用的设备为P.E 2400元素分析仪2400Ⅱ型。
原子吸收分析使用的设备为SHIMADZU,AA-7000型。
原子吸收分析条件为标准曲线法，方法采用常规火焰法。
对合成的黄芩苷镁进行元素分析和原子吸收分析，结果列于下表4中。
表4：合成黄芩苷镁元素分析和原子吸收分析结果
Figure PCTCN2017071616-appb-000006
以上结果均表明：Mg2+与两个黄芩苷的葡萄糖醛酸上的羧基发生反应，分子量为914，分子式为Mg(C21H16O11)2。
本发明还涉及所述的黄芩苷镁化合物的用途。在含黄芩或黄芩苷的中药单方或复方制剂中，可以用以同等剂量的黄芩苷镁化合物代替其中的黄芩或黄芩苷，或者它在制备用于治疗肝损伤、脑缺血、糖尿病、炎症或肿瘤药物中的用途。
本发明黄芩苷镁制备方法在制备过程中无需加酸碱，不会破坏黄芩中黄芩苷原有的存在形式，没有使用有毒有害有机试剂，绿色环保，产物纯度高。
经紫外光谱，红外光谱，原子吸收光谱法，核磁共振氢谱，碳谱，液质联用技术鉴定结构，证明从黄芩药材中提取的黄芩苷镁化合物与市售黄芩苷经过一定反应而得黄芩苷镁化合物两者结构一致。药代动力学 实验表明黄芩苷镁口服吸收速度比黄芩苷快，生物利用度比黄芩苷高。药理实验表明，所述的黄芩苷镁有较好的药理活性。
[有益效果]
本发明的有益效果是：本发明黄芩苷镁提取方法不会破坏黄芩中黄芩苷原有的存在形式，绿色环保，产物纯度高；合成方法简便，质量可控，能还原黄芩苷在黄芩的原有存在形式。黄芩苷镁相比黄芩苷溶解度大大提高，能使黄芩苷的吸收速率变快，口服生物利用度提高，能代替黄芩苷入药，还可以非常方便制成水溶性注射剂或粉针剂。药理实验表明，所述的黄芩苷镁有良好的药理活性。
【附图说明】
图1是酵母致热大鼠体温变化图。
图2是黄芩苷对酵母致热大鼠的干预作用图。
【具体实施方式】
通过下述实施例将能够更好地理解本发明。
实施例1：制备黄芩苷镁化合物
该实施例的实施步骤如下:
A、制备黄芩苷混悬液
按照以克计黄芩苷与以毫升计纯化水的比为1:30，把黄芩苷粉加到纯化水中，混合均匀，得到一种黄芩苷混悬液；
B、制备含镁离子的黄芩苷混悬液
按照黄芩苷与镁的摩尔比1.0:1.0，往步骤A得到的黄芩苷混悬液中添加氢氧化镁镁化合物，混合均匀，得到一种含镁离子的黄芩苷混悬液；
C、反应
在搅拌的条件下，让步骤B得到的含镁离子的黄芩苷混悬液在温度30℃下进行反应，直到其反应体系变得澄清透明，使用布氏漏斗减压抽滤设备过滤，得到滤液；
D、干燥
让步骤C得到的滤液在由吴江华飞电热设备有限公司以商品名 HF881-2干燥箱的设备中以60℃中进行加热蒸干，得到水含量为以重量计1.0％以下的黄芩苷镁化合物。
实施例2：制备黄芩苷镁化合物
该实施例的实施步骤如下:
A、制备黄芩苷混悬液
按照以克计黄芩苷与以毫升计纯化水的比为1:50，把黄芩苷粉加到纯化水中，混合均匀，得到一种黄芩苷混悬液；
B、制备含镁离子的黄芩苷混悬液
按照黄芩苷与镁的摩尔比2.0:1.0，往步骤A得到的黄芩苷混悬液中添加氧化镁镁化合物，混合均匀，得到一种含镁离子的黄芩苷混悬液；
C、反应
在搅拌的条件下，让步骤B得到的含镁离子的黄芩苷混悬液在温度70℃下进行反应，直到其反应体系变得澄清透明，使用布氏漏斗减压抽滤设备过滤，得到滤液；
D、干燥
让步骤C得到的滤液使用由江苏锡山市林州干燥机厂公司以商品名QZR-5型销售的喷雾干燥设备进行喷雾干燥，得到水含量为以重量计1.0％以下的黄芩苷镁化合物。
实施例3：制备黄芩苷镁化合物
该实施例的实施步骤如下:
A、制备黄芩苷混悬液
按照以克计黄芩苷与以毫升计纯化水的比为1:20，把黄芩苷粉加到纯化水中，混合均匀，得到一种黄芩苷混悬液；
B、制备含镁离子的黄芩苷混悬液
按照黄芩苷与镁的摩尔比0.5:1.0，往步骤A得到的黄芩苷混悬液中添加碱式碳酸镁镁化合物，混合均匀，得到一种含镁离子的黄芩苷混悬液；
C、反应
在搅拌的条件下，让步骤B得到的含镁离子的黄芩苷混悬液在温度60℃下进行反应，直到其反应体系变得澄清透明，使用布氏漏斗减压抽滤设备过滤，得到滤液；
D、干燥
让步骤C得到的滤液使用由北京五洲东方科技发展有限公司以商品名EYELA OSB-2100型的冷冻干燥设备在-40℃条件下进行冷冻干燥，得到水含量为以重量计1.0％以下的黄芩苷镁化合物。
实施例4：制备黄芩苷镁化合物
该实施例的实施步骤如下:
A、制备黄芩苷混悬液
按照以克计黄芩苷与以毫升计纯化水的比为1:20，把黄芩苷粉加到纯化水中，混合均匀，得到一种黄芩苷混悬液；
B、制备含镁离子的黄芩苷混悬液
按照黄芩苷与镁的摩尔比0.5:1.0，往步骤A得到的黄芩苷混悬液中添加硫酸镁镁化合物，混合均匀，得到一种含镁离子的黄芩苷混悬液；
C、反应
在搅拌的条件下，让步骤B得到的含镁离子混悬液在温度70℃下进行反应，直到其反应体系变得澄清透明，使用布氏漏斗减压抽滤设备过滤，得到滤液；
D、干燥
让步骤C得到的滤液使用由江苏锡山市林州干燥机厂公司以商品名QZR-5型销售的喷雾干燥设备进行喷雾干燥，得到水含量为以重量计1.0％以下的黄芩苷镁化合物。
实施例5：制备黄芩苷镁化合物
该实施例的实施步骤如下:
A、制备黄芩苷混悬液
按照以克计黄芩苷与以毫升计纯化水的比为1:100，把黄芩苷粉加到纯化水中，混合均匀，得到一种黄芩苷混悬液；
B、制备含镁离子的黄芩苷混悬液
按照黄芩苷与镁的摩尔比2.0:1.0，往步骤A得到的黄芩苷混悬液中添加硝酸镁镁化合物，混合均匀，得到一种含镁离子的黄芩苷混悬液；
C、反应
在搅拌的条件下，让步骤B得到的含镁离子混悬液在温度60℃下进行反应，直到其反应体系变得澄清透明，使用布氏漏斗减压抽滤设备过滤，得到滤液；
D、干燥
让步骤C得到的滤液使用由北京五洲东方科技发展有限公司以商品名EYELA OSB-2100型销售的冷冻干燥设备在-40℃条件下进行冷冻干燥，得到水含量为以重量计1.0％以下的黄芩苷镁化合物。
实施例6：制备黄芩苷镁化合物
该实施例的实施步骤如下:
A、制备黄芩苷混悬液
按照以克计黄芩苷与以毫升计纯化水的比为1:30，把黄芩苷粉加到纯化水中，混合均匀，得到一种黄芩苷混悬液；
B、制备含镁离子的黄芩苷混悬液
按照黄芩苷与镁的摩尔比1.0:1.0，往步骤A得到的黄芩苷混悬液中添加氯化镁镁化合物，混合均匀，得到一种含镁离子的黄芩苷混悬液；
C、反应
在搅拌的条件下，让步骤B得到的含镁离子混悬液在温度50℃下进行反应，直到其反应体系变得澄清透明，使用布氏漏斗减压抽滤设备过滤，得到滤液；
D、干燥
让步骤C得到的滤液使用由北京五洲东方科技发展有限公司以商品名EYELA OSB-2100型销售的冷冻干燥设备在-40℃条件下进行冷冻干燥，得到水含量为以重量计1.0％以下的黄芩苷镁化合物。
实施例7：制备黄芩苷镁化合物
该制备方法的步骤如下：
A、大孔吸附树脂预处理
将由郑州勤实科技有限公司以商品名HPD-100型销售的大孔吸附树脂在浓度以体积计95％乙醇水溶液中浸泡26小时，然后湿法装柱，接着让浓度以体积计95％乙醇水溶液以1.5BV/h流速通过该大孔吸附树脂柱，直到流出液与水按照体积比1:5混合后不再呈现白色浑浊状，接着用蒸馏水以1.7BV/h流速洗脱至无色；
B、提取
按照以克计黄芩与以ml计乙醇水溶液的比为1:11，把黄芩加到浓度以体积计50％乙醇水溶液中，在温度60℃与搅拌的条件下提取1.0小时，然后分离，得到一种提取液与一种残留物；所述的残留物在与上述相同条件下重复提取2次，提余残留物弃去；提取液合并，得到一种黄芩提取液；
C、吸附与洗脱
让步骤B得到的黄芩提取液以1.5BV/h流速通过步骤A预处理的大孔吸附树脂柱进行吸附，然后使用4个大孔吸附树脂床体积水洗涤，洗涤水弃去；再用4个大孔吸附树脂床体积浓度为以体积计50％的乙醇水溶液洗脱，收集含有黄芩苷的洗脱液；
D、浓缩与干燥
将步骤C得到的含有黄芩苷的洗脱液，使用由上海申顺生物科技有限公司以商品名R2002旋转蒸发器销售的设备在压力0.01MPa与温度50℃条件下减压浓缩，直至浓缩液的体积为以克计黄芩药材重量的1倍，再在由吴江华飞电热设备有限公司以商品名HF881-2干燥箱的设备中以60℃进行加热蒸干，得到一种黄芩苷镁化合物粗提物；
E、纯化
对步骤D得到黄芩苷镁化合物粗提物在80％乙醇溶剂中在60℃条件下重结晶纯化，得到所述的黄芩苷镁化合物。
实施例8：制备黄芩苷镁化合物
该制备方法的步骤如下：
A、大孔吸附树脂预处理
将由郑州勤实科技有限公司以商品名AB-8型销售的大孔吸附树脂在浓度以体积计95％乙醇水溶液中浸泡20小时，然后湿法装柱，接着让浓度以体积计95％乙醇水溶液以2.5BV/h流速通过该大孔吸附树脂柱，直到流出液与水按照体积比1:5混合后不再呈现白色浑浊状，接着用蒸馏水以1.5BV/h流速洗脱至无色；
B、提取
按照以克计黄芩与以ml计乙醇水溶液的比为1:8，把黄芩加到浓度以体积计40％乙醇水溶液中，在温度55℃与搅拌的条件下提取1.5小时，然后分离，得到一种提取液与一种残留物；所述的残留物在与上述相同条件下重复提取3次，提余残留物弃去；提取液合并，得到一种黄芩提取液；
C、吸附与洗脱
让步骤B得到的黄芩提取液以2.5BV/h流速通过步骤A预处理的大孔吸附树脂柱进行吸附，然后使用5个大孔吸附树脂床体积水洗涤，洗涤水弃去；再用5个大孔吸附树脂床体积浓度为以体积计45％的乙醇水溶液洗脱，收集含有黄芩苷的洗脱液；
D、浓缩与干燥
将步骤C得到的含有黄芩苷的洗脱液，使用由上海申顺生物科技有限公司以商品名R2002旋转蒸发器销售的设备在压力0.03MPa与温度35℃条件下减压浓缩，直至浓缩液的体积为以克计黄芩药材重量的15倍，再使用由江苏锡山市林州干燥机厂公司以商品名QZR-5型销售的喷雾干燥设备喷雾干燥，得到一种黄芩苷镁化合物粗提物；
E、纯化
对步骤D得到黄芩苷镁化合物粗提物使用由YMC公司以商品名GEL C18AAG12S50销售的ODS柱在10％～40％甲醇水作为洗脱液条件下进行纯化，得到所述的黄芩苷镁化合物。
实施例9：制备黄芩苷镁化合物
该制备方法的步骤如下：
A、大孔吸附树脂预处理
将由郑州西电电力树脂销售有限公司以商品名D-101型销售的大孔吸附树脂在浓度以体积计95％乙醇水溶液中浸泡28小时，然后湿法装柱，接着让浓度以体积计95％乙醇水溶液以2.2BV/h流速通过该大孔吸附树脂柱，直到流出液与水按照体积比1:5混合后不再呈现白色浑浊状，接着用蒸馏水以2.0BV/h流速洗脱至无色；
B、提取
按照以克计黄芩与以ml计乙醇水溶液的比为1:12，把黄芩加到浓度以体积计60％乙醇水溶液中，在温度65℃与搅拌的条件下提取1.0小时，然后分离，得到一种提取液与一种残留物；所述的残留物在与上述相同条件下重复提取2次，提余残留物弃去；提取液合并，得到一种黄芩提取液；
C、吸附与洗脱
让步骤B得到的黄芩提取液以2.0BV/h流速通过步骤A预处理的大孔吸附树脂柱进行吸附，然后使用4个大孔吸附树脂床体积水洗涤，洗涤水弃去；再用6个大孔吸附树脂床体积浓度为以体积计55％的乙醇水溶液洗脱，收集含有黄芩苷的洗脱液；
D、浓缩与干燥
将步骤C得到的含有黄芩苷的洗脱液，使用由上海申顺生物科技有限公司以商品名R2002旋转蒸发器销售的设备在压力0.1MPa与温度65℃条件下减压浓缩，直至浓缩液的体积为以克计黄芩药材重量的5倍，再使用由北京四环科学仪器厂有限公司以商品名LGJ-22D型销售的冷冻干燥设备在-40℃条件下冷冻干燥，得到一种黄芩苷镁化合物粗提物；
E、纯化
对步骤D得到黄芩苷镁化合物粗提物使用由博纳艾杰尔公司以商品名Innoval C18销售的制备液相色谱柱在甲醇-水(30:70)色谱条件下 进行纯化，得到所述的黄芩苷镁化合物。
实施例10：制备黄芩苷镁化合物
该制备方法的步骤如下：
A、大孔吸附树脂预处理
将由郑州勤实科技有限公司以商品名YWD06B型销售的大孔吸附树脂在浓度以体积计95％乙醇水溶液中浸泡24小时，然后湿法装柱，接着让浓度以体积计95％乙醇水溶液以2.0BV/h流速通过该大孔吸附树脂柱，直到流出液与水按照体积比1:5混合后不再呈现白色浑浊状，接着用蒸馏水以2.5BV/h流速洗脱至无色；
B、提取
按照以克计黄芩与以ml计乙醇水溶液的比为1:9，把黄芩加到浓度以体积计45％乙醇水溶液中，在温度65℃与搅拌的条件下提取1.0小时，然后分离，得到一种提取液与一种残留物；所述的残留物在与上述相同条件下重复提取3次，提余残留物弃去；提取液合并，得到一种黄芩提取液；
C、吸附与洗脱
让步骤B得到的黄芩提取液以2.0BV/h流速通过步骤A预处理的大孔吸附树脂柱进行吸附，然后使用5个大孔吸附树脂床体积水洗涤，洗涤水弃去；再用5个大孔吸附树脂床体积浓度为以体积计50％的乙醇水溶液洗脱，收集含有黄芩苷的洗脱液；
D、浓缩与干燥
将步骤C得到的含有黄芩苷的洗脱液，使用由上海申顺生物科技有限公司以商品名R2002旋转蒸发器销售的设备在压力0.07MPa与温度45℃条件下减压浓缩，直至浓缩液的体积为以克计黄芩药材重量的10倍，再使用由江苏锡山市林州干燥机厂公司以商品名QZR-5型销售的喷雾干燥设备喷雾干燥，得到一种黄芩苷镁化合物粗提物；
E、纯化
对步骤D得到黄芩苷镁化合物粗提物使用由YMC公司以商品名 GEL C18AAG12S50销售的ODS柱在10％～40％甲醇水作为洗脱液条件下进行纯化，得到所述的黄芩苷镁化合物。
药理试验1：对肝损伤的保护作用
取健康小鼠40只，随机分为4组，分别为正常组、模型组、黄芩苷组、黄芩苷镁组。黄芩苷组给药剂量10mg/kg，黄芩苷镁组给药剂量相当于10mg/kg黄芩苷，正常组，模型组以相同体积生理盐水灌胃，共灌胃5天。在第4天，模型组和给药组腹腔注射D-半乳糖0.5g/kg，正常组腹腔注射相同体积生理盐水。检测血清ALT、AST、SOD、GSH-Px的活性及MDA的含量。应用SPSS-19.0统计软件进行方差分析。其结果列于表5中。
表5：对肝损伤保护作用结果
Figure PCTCN2017071616-appb-000007
*与模型组比较P<0.05；Δ与黄芩苷组比较P<0.05
表5的结果表明两种黄芩苷对D-半乳糖所致急性肝损伤小鼠血清ALT、AST的升高有显著降低作用，同时升高SOD、GSH-Px的活力，降低MDA的含量(P<0.05)；与黄芩苷组相比，黄芩苷镁组对D-半乳糖所致急性肝损伤小鼠小鼠血清ALT、AST升高的降低作用更显著，同时升高SOD、GSH-Px，降低MDA的含量更显著(P<0.05)。
药理试验2：对脑缺血小鼠学习记忆功能的影响
健康小鼠80只，随机分4组，分别为模型组，假手术组，黄芩苷组，黄芩苷镁组，每组20只。
各组的处理如下：
模型组：用1％的戊巴比妥钠腹腔注射麻醉小鼠，麻醉后暴露分离左右两侧颈总动脉，用血管钳迅速分别夹闭两侧动脉各20min，松开血管钳，缝合，放回笼中饲养；假手术组：相同方法暴露左右两侧颈总动脉，但不夹闭，缝合伤口，放回笼中饲养；黄芩苷组：操作同模型组， 手术后第2天给予腹腔注射12.5mg/kg黄芩苷，连续注射12天；黄芩苷镁组：操作同模型组，手术后第2天给予腹腔注射黄芩苷镁(相当于黄芩苷12.5mg/kg)，连续注射12d，假手术组与模型组分别给予等体积的生理盐水。
水迷宫：测试用圆形水池直径120cm，高60cm，圆柱形平台直径10cm，高40cm，水池内部均为黑色。上方安置连接显示器的摄像机，记录小鼠的运动轨迹。实验时水深12cm，水温(25±2)℃，迷宫周围一切空间标志在整个实验过程中保持不变。
水迷宫记忆测试实验：正式实验开始前1天，让每只小鼠在迷宫中熟悉水环境2min，第2天开始正式实验。将小鼠面向并靠近池壁随机从四个不同象限(North，West，South，East，为一个时段)入水，记录下小鼠入水到爬上平台的时间，此为逃避潜伏期。在120秒内未找到平台的，潜伏期记为120秒，同时引导其找到平台并休息60秒。连续训练15天，2次/天。各组动物依次进行水迷宫实验，第1天的路经包括2个盲端，第2天有3个盲端，第3、4、5天均为4个盲端。小鼠面向池壁在起始点放入水中，记录小鼠放入水中到达终点平台时间(潜伏期)和进入盲端的次数。如小鼠在60秒内未到达平台，可将小鼠赶至平台，并令小鼠在平台上呆20秒，每日上下午各训练1次，实验5天。
实验数据以均数±标准差(x±s)表示，采用SPSS19.0统计软件对数据进行单因素方差分析，P＜0.05为差异有统计学意义。
表6：抗脑缺血小鼠学习记忆功能损伤的保护作用
Figure PCTCN2017071616-appb-000008
注：*与模型组比较P<0.05；Δ与黄芩苷组比较P<0.05
表6的结果表明黄芩苷组与黄芩苷镁组小鼠从水迷宫训练学习的逃避潜伏期显著缩短、次数减少，与模型组相比差异有统计学意义，黄芩苷镁组与黄芩苷组相比，逃避潜伏期显著缩短、次数减少，差异有统计 学意义。
药理试验3：抗炎活性
取健康小鼠144只，随机分成3组。分别为黄芩苷组、黄芩苷镁组及对照组，每大组按时间点1，3，5，7，11，15h又分为6个小组，每组8只。黄芩苷组剂量100mg·kg-1、黄芩苷镁剂量相当于100mg·kg-1黄芩苷，口服灌胃给药，对照组给予同体积生理盐水。每组在每个时间点到达前40min给小鼠左耳滴加100微升二甲苯，时间到达时，处死动物并用打孔器取小鼠两耳的耳片，以右耳与左耳的重量差(肿胀度)为指标观察药物的抗炎作用，结果见表7。
表7：黄芩苷与黄芩苷镁抑制二甲苯致小鼠耳廓肿胀的时效关系(n＝8)
Figure PCTCN2017071616-appb-000009
注:*与正常对照组(10.25±3.15)比较，P<0.05，Δ与黄芩苷组比较，P<0.05
表7的结果表明黄芩苷组与黄芩苷镁组均可显著对抗二甲苯致小鼠耳廓肿胀炎症反应；同时黄芩苷镁组与黄芩苷组在给药后1h抗炎作用已经表现出显著的差异，黄芩苷镁组强于黄芩苷组；结果表明时黄芩苷镁抗炎作用出现早，作用平稳，维持时间较长，表明其吸收起效快。
药理试验4：抗肿瘤活性
取人肝癌HepG2细胞，用含10％小牛血清、青霉素(100μmol·L-1)、链霉素(1mg·mL-1)的DMEM培养基，置37℃，5％CO2孵箱内培养。
取对数生长期的HepG2细胞，用含浓度10％的胎牛血清RPMI-1640培养基稀释至菌液浓度为1×104个/mL，接种于96孔板中，每孔100μL，在温度37℃、相对湿度90％与5％CO2培养箱内培养，培养24h后弃去培养液，分别加入黄芩苷和黄芩苷镁(相当于黄芩苷浓度(5，10，20μg/mL))含药培养液，每个药物浓度设4个平行复孔，与试验孔平行 操作不加细胞只加培养液的空白对照孔，只加细胞不加药物的阴性对照组，继续培养48h，收集细胞。用PBS液洗涤2次后，采用流式细胞仪FACS检测，并进行参数获取和数据分析，其结果列于表8中。
表8：黄芩苷与黄芩苷镁对人肝癌HepG2细胞凋亡影响
Figure PCTCN2017071616-appb-000010
注:*与对照组比较，P<0.05，Δ与黄芩苷组比较，P<0.05
表8的结果表明肝癌细胞HepG2分别在浓度为5，10，20μg/mL的黄芩苷溶液和黄芩苷镁溶液中作用48h，均有不同程度的凋亡；相同浓度的黄芩苷镁对细胞凋亡的影响明显强于黄芩苷，结果具有统计学差异。
药理试验5：对小鼠镇痛作用
取40只小鼠，体重20～22g，随机分为4组，分别为黄芩苷0.067g/kg组，黄芩苷镁(相当于黄芩苷0.067g/kg)，空白对照组(给予生理盐水)，
阿斯匹林阳性对照组(0.15g/kg)，每组10只。每天灌胃给药1次，连续7天。于末次给药后30min，按照0.2mL/10g腹腔注射0.6％醋酸生理盐水溶液，观察15min内小鼠出现扭体反应次数，计算镇痛率，镇痛率＝[(空白对照组平均扭体次数－实验组平均扭体次数)/空白对照组平均扭体次数]×100％。采用SPSS19.0统计软件对数据进行单因素方差分析，P＜0.05为差异有统计学意义，结果见表9。
表9：黄芩苷和黄芩苷镁对醋酸所致小鼠疼痛作用的影响
Figure PCTCN2017071616-appb-000011
注:*与对照组比较，P<0.05，Δ与黄芩苷组比较，P<0.05
表9的结果表明与空白对照组比较，黄芩苷镁组对醋酸所致小鼠疼痛的反应有明显的抑制作用，且效果优于黄芩苷组。
药理试验6：解热作用
取雄性SD大鼠，随机分为4组，分别为对照组、模型组、黄芩苷组和黄芩苷镁组。动物饲养环境为常规日夜周期，室温(22±2)℃，相对湿度(50±2)％。实验前3天将大鼠置于实验环境中摸拟操作使其适应实验条件，选取基础体温在37.5～39.0℃的大鼠，第4天开始实验。实验前禁食12h，实验开始后每隔1h测1次体温，取3次平均值作为基础体温，3,次体温中最高值与最低值之差大于0.5℃者剔除。
先灌胃给予黄芩苷或黄芩苷镁，然后立即皮下注射20％干酵母生理盐水混悬液(对照组除外)。给药方式如下：对照组灌胃2ml/kg生理盐水+皮下注射生理盐水10ml/kg，模型组灌胃2ml/kg生理盐水+皮下注射酵母2g/kg，黄芩苷组灌胃80mg/kg+皮下注射酵母2g/kg，黄芩苷镁组灌胃(相当于80mg/kg黄芩苷)+皮下注射酵母2g/kg，给药后，每2h测体温1次，观察注射酵母后不同时间的体温变化(ΔT，℃)，连续测量14h，其结果列于附图1与附图2中。
图1的结果表明模型组大鼠皮下注射酵母混悬液后8h体温达高峰，ΔT为1.4℃，作用维持14h以上。图2表明对照组动物的体温在实验过程中无明显变化。模型组大鼠皮下注射酵母混悬液后6h和8h的ΔT分别为0.95，1.4℃，与对照组比较，体温升高有显著性意义(P<0.05)。6h时黄芩苷、黄芩苷镁组的ΔT分别为0.4，0.2℃，与模型组同时间比均明显降低(P<0.05)，8h黄芩苷、黄芩苷镁组ΔT分别为0.8，0.3℃，与模型组相比也均明显降低(P<0.05)；同时黄芩苷镁与黄芩苷比较，6h和8h时ΔT为0.2，0.5℃，结果表明黄芩苷对酵母诱导的发热具有抑制或干预作用，同时黄芩苷镁的效果更好。
药理试验7：抗糖尿病疾病
取健康SD大鼠，雄性，6～8周龄，随机分为4组，分别为高脂饲料对照组，糖尿病对照组、黄芩苷治疗组和黄芩苷镁治疗组。预养1周。 第2周开始全部大鼠饲喂高脂饲料，6周过后，大鼠禁食不禁水12h，将糖尿病对照组、黄芩苷治疗组和黄芩苷镁治疗组大鼠分别尾静脉注射用柠檬酸缓冲液(pH4.5)配制的链脲佐菌素45mg/kg，3～7天后测空腹血糖，血糖值＞13mmol/L，为糖尿病鼠模型成功的标准。高脂饲料对照组尾静脉注射给予柠檬酸缓冲液。
造模成功后开始给药，每天上午开始灌胃给予药物。黄芩苷组剂量为80mg/kg和黄芩苷镁组(相当于黄芩苷80mg/kg)，糖尿病模型和高脂饲料对照组大鼠灌胃给予同体积的溶剂，每天1次，给药6周。造模后及给药后第3和第6周测血糖。采用SPSS19.0统计软件对数据进行单因素方差分析，P＜0.05为差异有统计学意义，结果见表10。
表10：黄芩苷和黄芩苷镁对链脲佐菌素糖尿病大鼠血糖的影响
Figure PCTCN2017071616-appb-000012
注:*与对照组比较，P<0.05，Δ与黄芩苷组比较，P<0.05，#与模型组比较，P<0.05
表10的结果表明实验第3周时，糖尿病模型组大鼠血糖升高，黄芩苷组和黄芩苷镁组血糖分别下降。第6周时，糖尿病模型组大鼠血糖仍然在升高，而黄芩苷组和黄芩苷镁组的血糖进一步下降，黄芩苷和黄芩苷镁组血糖值明显低于糖尿病模型组(P＜0.05)，表明黄芩苷能改善糖尿病症状。同时黄芩苷镁组血糖降低程度明显强于黄芩苷组(P＜0.05)。
试验实施例：黄芩苷与黄芩苷镁化合物在水中溶解度的比较
采用常规溶解度测定方法，在温度37℃水中，测定了黄芩苷与黄芩苷镁的溶解度，其结果列于表11中。
表11：黄芩苷与黄芩苷镁化合物在水中的溶解度
Figure PCTCN2017071616-appb-000013
表11的结果表明黄芩苷镁化合物的水溶性较好，是黄芩苷的2225倍。

Claims (10)

  1. 一种黄芩苷镁化合物，其特征在于它是具有下述化学结构式(I)的化合物：
    Figure PCTCN2017071616-appb-100001
  2. 根据权利要求1所述黄芩苷镁化合物的制备方法，其特征在于该制备方法的步骤如下:
    A、制备黄芩苷混悬液
    按照以克计黄芩苷与以毫升计纯化水的比为1:20～100，把黄芩苷粉加到纯化水中，混合均匀，得到一种黄芩苷混悬液；
    B、制备含镁离子的黄芩苷混悬液
    按照黄芩苷与镁的摩尔比0.5～3.0:1，往步骤A得到的黄芩苷混悬液中添加镁化合物，混合均匀，得到一种含镁离子的黄芩苷混悬液；
    C、反应
    在搅拌的条件下，让步骤B得到的含镁离子的黄芩苷混悬液在温度20～70℃下进行反应，直到其反应体系变得澄清透明，过滤，得到滤液；
    D、干燥
    将步骤C得到的滤液进行干燥，得到所述的黄芩苷镁化合物。
  3. 根据权利要求2所述的制备方法，其特征在于所述的镁化合物选自氢氧化镁、氧化镁、碱式碳酸镁、乙酸镁、硫酸镁、硝酸镁或氯化镁。
  4. 根据权利要求1所述黄芩苷镁化合物的制备方法，其特征在于该制备方法的步骤如下：
    A、大孔吸附树脂预处理
    将大孔吸附树脂在浓度以体积计95％乙醇水溶液中浸泡20～28小时，然后湿法装柱，接着让浓度以体积计95％乙醇水溶液以1.5～2.5BV/h流速通过该大孔吸附树脂柱，直到流出液与水按照体积比1:5混合后不再呈现白色浑浊状，接着用蒸馏水以1.5～2.5BV/h流速洗脱至无色；
    B、提取
    按照以克计黄芩与以毫升计乙醇水溶液的比为1:8～15，把黄芩加到浓度以体积计40～60％乙醇水溶液中，在温度55～65℃与搅拌的条件下提取0.8～1.2小时，然后分离，得到一种提取液与一种残留物；所述的残留物在与上述相同条件下重复提取2～3次，提取残留物弃去；提取液合并，得到一种黄芩提取液；
    C、吸附与洗脱
    让步骤B得到的黄芩提取液以1.5～2.5BV/h流速通过步骤A预处理的大孔吸附树脂柱进行吸附，然后使用4～6个大孔吸附树脂床体积水洗涤，洗涤水弃去；再用4～6个大孔吸附树脂床体积浓度为以体积计45～55％的乙醇水溶液洗脱，收集含有黄芩苷的洗脱液；
    D、浓缩与干燥
    将步骤C得到的含有黄芩苷的洗脱液，在温度35～65℃条件下减压浓缩，直至浓缩液的体积为以克计黄芩药材重量的1～15倍，再经干燥得到一种黄芩苷镁化合物粗提物；
    E、纯化
    对步骤D得到黄芩苷镁化合物粗提物进行纯化，得到所述的黄芩苷镁化合物。
  5. 根据权利要求4所述的制备方法，其特征在于所述的大孔吸 附树脂选自HPD-100、AB-8、D101或YWD06B。
  6. 根据权利要求4所述的制备方法，其特征在于所述大孔吸附树脂的粒度10～80目。
  7. 根据权利要求4所述的制备方法，其特征在于所述大孔吸附树脂柱的直径与柱高之比是1：3～8。
  8. 根据权利要求2或4所述的制备方法，其特征在于所述的干燥是加热蒸干、喷雾干燥或冷冻干燥。
  9. 根据权利要求2或4所述的制备方法，其特征在于得到的黄芩苷镁化合物采用重结晶、十八烷基硅烷反相柱、制备液相色谱法进行纯化，以达到黄芩苷镁化合物含量为以重量计95％以上。
  10. 根据权利要求1所述的黄芩苷镁化合物的用途，其特征在于所述的黄芩苷镁化合物以等剂量代替在含黄芩或黄芩苷的中药单方或复方制剂中的黄芩或黄芩苷，或者它在制备用于治疗肝损伤、脑缺血、糖尿病、炎症或肿瘤药物中的用途。
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