WO2012126354A1 - Baicalin-copper complex, preparation method and application thereof - Google Patents

Abstract

The present invention relates to a baicalin-copper complex, a preparation method and an application thereof, relating to the field of pharmacology. The present invention utilizes different ingredients and different molar ratios in a reaction to produce two baicalin-copper complexes with new structures, and uses the complexes to conduct in vitro antibacterial experiments. The results of the experiments show that the complexes have strong antibacterial properties. In comparison with baicalin-metal complexes in the prior art, the present complex is more effective and can be used as a feed additive, and as a drug for preparing antibacterial agents, especially anti-drug resistant antibacterial agents. The present invention also relates to a method for preparing baicalin-metal complexes, which is simple, easy to control, and produces a highly pure final product.

Description

 Instruction manual

 Astragalus copper complex and preparation method and application thereof

Technical field

 The invention relates to the field of chemical medicines, in particular to a yellow bismuth copper complex and a preparation method thereof. Background technique

 Baicalin is an important natural medicine and active ingredient of traditional Chinese medicine. It has extremely wide biological activity and has antibacterial, hypoglycemic, antiviral, anti-inflammatory, diuretic, anti-tumor, antioxidant free radicals, anti-allergic reaction and antispasmodic. a pharmacological effect.

 Fe and Cu are the essential trace elements in the human body, which are ranked first and second respectively. There are many important enzymes in the body containing Cu. Cu activates the formation of hemoglobin, promotes the absorption and utilization of iron, and has a role in the transmission of electrons, the synthesis of elastin, the metabolism of connective tissue, the metabolism of sputum, the formation of phospholipids and nerve tissue. Significance.

 Many studies have shown that the chelation of baicalin with a proper amount of metal ions can increase its biological effects. The root cause may be: 1 It is easier to contribute electrons to oxidative radicals than the original ligands, thereby exerting antioxidant effects; The structure is similar to many enzymes with metal as the active center, and can function in combination with such enzymes; 3 The complex aggregates the synergistic action of ligand and metal to produce 1+1>2 pharmacological activity.

 Many medicinal chemists have performed extensive structural modification of baicalin to obtain modified structures with higher activity or chemical properties that are more beneficial to human body absorption.

 Jia Zhaoxia, Wu Hao, etc. (See "Inorganic Chemistry Journal, 1990, 6 (1): 106-108") Dissolving baicalin in 50% methanol solution under alkaline conditions of sodium hydroxide (pH=7.4) The reaction was carried out with copper chloride to obtain a brown needle-copper saponin (CuB), which was washed with 60% methanol and dried, and its structure was as shown in the formula (I).

张素俊等 （参见"南京中医学院学报， 1991， 7 (4) :235-236") 将黄芩苷溶于无水 乙醇溶液中， 在碳酸钠碱性条件下， 加入醋酸铜反应， 回流得到棕红色粉末， 用热无水 乙醇洗涤后干燥， 其结构

Zhang Sujun et al. (See "Journal of Nanjing University of Traditional Chinese Medicine, 1991, 7 (4): 235-236"). Dissolve baicalin in an anhydrous ethanol solution, add copper acetate under alkaline conditions, and return to brownish red. Powder, dried with hot anhydrous ethanol and dried, its structure

 II

It can be seen from the above two published documents that the same copper ion reacts with baicalin, and the structure of the obtained complex is completely different, which has a great relationship with the copper ion-containing compound and its amount. However, the above two reactions all dissolve baicalin in an alcohol solution, and react the copper ion-containing compound with baicalin under weak alkaline conditions, but under alkaline conditions, a large amount of Cu (OH) is generated. ₂ precipitation, the precipitate will precipitate together with the obtained complex, and it can not be cleaned by washing with alcohol. Therefore, the complex obtained by the methods disclosed in the above two documents is actually a mixture and cannot be pure. It is difficult to separate the complex from the hydroxide precipitate, so purification of the final product is difficult. Existing complexes involving the formation of baicalin with other metals have reported similar difficulties and disadvantages in the synthesis and purification.

 Because there are many active groups (ie, coordination sites) in the coordination reaction of the baicalin with the metal, various complex structures can be formed, and in combination with the pharmacological activity of baicalin, it is desired to obtain more different effects or A higher-yield baicalin metal complex.

Summary of the invention

 In view of the deficiencies in the above-mentioned fields, the present invention provides a novel structure of a xanthine copper complex, which has a strong antibacterial function in vitro, and has a better medicinal effect than the conventionally disclosed xanthine copper complex. .

 At the same time, the invention also provides a preparation method of a yellow bismuth copper complex, which is simple and easy to control, and has high purity of the final product.

A yellow copper complex, whose structure is as shown in the formula (ΠΙ):

 III

 A method for preparing a xanthine copper complex represented by formula (m), characterized in that: baicalin, copper sulfate or copper chloride is dissolved in a non-aqueous solution, respectively, and the molar ratio is 2:1, 1:1 or The 1:2 mixed reaction gave a brownish yellow precipitate, which was filtered, washed several times with ethanol, and dried. The non-aqueous solution was one or more of methanol, ethanol, ethyl acetate and petroleum ether.

 The reaction conditions are: a reaction temperature of 30 to 80 ° C and a reaction time of 2-3 hours.

 Astragalus copper complex, its

 IV

 A method for preparing a xanthine copper complex represented by formula (IV), characterized in that: baicalin and copper acetate are respectively dissolved in a non-aqueous solution, and the molar ratio of the two is 2:1 or 1:1 to obtain a brown-red precipitate. It is filtered, washed several times with ethanol, and dried. The non-aqueous solution is one or more of methanol, ethanol, ethyl acetate and petroleum ether.

 The reaction conditions are: a reaction temperature of 30 to 80 ° C and a reaction time of 2-3 hours.

 The above-mentioned yellow yttrium copper complex is used as a servo additive.

 The use of the above-mentioned xanthine copper complex in the preparation of an antibacterial or antitumor drug.

 The antibacterial agent refers to a drug resistant to a drug, the drug-resistant bacteria refers to a drug-resistant pig-derived Escherichia coli; a drug-resistant Salmonella typhimurium; a drug-resistant Staphylococcus aureus.

 The anti-tumor drug refers to a drug against lung cancer or anti-liver cancer.

 An antibacterial agent characterized in that the antibacterial active substance comprises the above-mentioned xanthine copper complex.

The antibacterial agent further includes a traditional Chinese medicine, a western medicine or the like which is compatible with the yellow scorpion copper complex. Antibacterial and harmless chemical composition.

 The medicament further comprises a pharmaceutically acceptable auxiliary ingredient.

 The medicament is in the form of a capsule, an injection, a tablet, a granule, an oral solution, an ointment, a powder or a suspension. An antitumor drug characterized in that the antitumor active substance comprises the above-mentioned xanthine copper complex. The anti-tumor drug further includes a traditional Chinese medicine, a western medicine or a harmless chemical component which is beneficial for anti-tumor, which is compatible with the yellow scorpion copper complex.

 The antitumor drug further comprising a pharmaceutically acceptable auxiliary ingredient.

 The antitumor drug is in the form of a capsule, an injection, a tablet, a granule, an oral solution, an ointment, a powder or a suspension.

 The invention divides the experiment into two groups:

 (1) Dissolving baicalin, copper sulfate or copper chloride in a non-aqueous solution separately, reacting at a certain molar ratio (2:1, 1:1, 1:2) at a certain temperature for 2-3 hours. The precipitate was brownish yellow, filtered, washed with ethanol several times, and dried to give a brownish brown solid. The non-aqueous solution is one or more of methanol, ethanol, ethyl acetate and petroleum ether, and the reaction temperature is 30-80 °C.

 (2) Dissolving baicalin and copper acetate in a non-aqueous solution separately, reacting at different molar ratios (2:1, 1:1) at the same temperature for 2-3 hours, obtaining a brown-red precipitate, filtering, using ethanol Washing several times, drying, to obtain a brownish red powdery solid B. The non-aqueous solution is one or more of methanol, ethanol, ethyl acetate and petroleum ether, and the reaction temperature is 30-80 °C.

 The above experiments prove that different molar ratios, different metal salts of the same metal and yellow baicalin may be obtained by the same structure of compounds, or may be compounds of different structures. The present invention uses the UV, Infra-red NMR products respectively. Structural characterization by LC-MS, metal element analysis, etc., and the product A obtained in the above group (1) is estimated to have the structural formula as shown in (III); the product B obtained in the group (2) has the structural formula (IV) Shown.

 The method of the invention dissolves the two raw materials separately and reacts at the original pH value, thereby avoiding the possibility of generating copper hydroxide under alkaline conditions, so the product obtained by the method of the invention has high purity and eliminates complicated Purification step.

 In the present invention, the above-mentioned partial product (complexes A and B obtained by reacting baicalin with metallic copper) is also subjected to an in vitro antibacterial test, which indicates that it has a high antibacterial effect and can be used as a servo additive.

 In the present invention, the obtained xanthine copper complex is subjected to an in vitro test against the drug resistant bacteria. The test results show that it has high antibacterial activity against drug-resistant Escherichia coli, drug-resistant Salmonella and drug-resistant Staphylococcus aureus. It can be used for anti-resistant bacteria, such as anti-resistant bacteria as an active ingredient.

In vitro studies of the present invention have found that two xanthine copper complexes have better inhibition of tumors than baicalin. Its role, especially lung cancer and liver cancer cells _; therefore, it can be used as an antitumor active ingredient for the preparation of antitumor drugs. detailed description

Example 1:

 1-1 synthetic complex

 The invention divides the experiment into two groups:

 (1) Dissolving baicalin, copper sulfate or copper chloride in a non-aqueous solution, respectively, in a ratio of different molar ratios (2:1, 1:1, 1:2) for 2-3 hours to obtain a brownish yellow precipitate. It was filtered, washed several times with ethanol, and dried to give a brownish-yellow solid A. The non-aqueous solution is one or more of methanol, ethanol, ethyl acetate and petroleum ether, and the reaction temperature is 30-80 °C.

 (2) Dissolving baicalin and copper acetate in a non-aqueous solution separately, and reacting for 2-3 hours at different molar ratios (2:1, 1:1). The products obtained by the reaction of two different molar ratios were brown-red precipitates, filtered, washed several times with ethanol, and dried to give a brownish red powdery solid B. The non-aqueous solution is one or more of methanol, ethanol, ethyl acetate and petroleum ether, and the reaction temperature is 30-80 °C.

 The synthetic yields of the above experiments are as follows: molar ratio 2:1 1:1 1:2 baicalin: copper chloride 46.37% 60.22% 46.13% baicalin: copper sulfate 46.73% 61.80% 45.78% baicalin: copper acetate 55.73% 74.62 %

1-2 Structural characterization

 UV Characterization of 1-2-1 Astragalus Copper Complex

1-2-1-1 Ultraviolet results of dissolution of dilute alkali solution

 The complex was dissolved in 2% NaOH to determine the maximum absorption wavelength. The results are shown in Table 1.

 Table 1 UV absorption of baicalin and its complexes in alkaline solution

1-2-1-2 DMSO溶解的紫外结果

UV results of 1-2-1-2 DMSO dissolution

 The complex was dissolved in DMSO and the maximum absorption wavelength was measured. The results are shown in Table 2.

 Table 2 UV absorption of baicalin and complex in DMSO solution

 Maximum absorption wavelength

 DMSO

 Baicalin 280 nm 316 nm

 Astragalus copper (all products) 292 nm —

 Maximum absorption wavelength change Red shift 12 nm —

 As can be seen from Tables 1 and 2, the product slightly differs from the UV absorption in DMSO in lye, and the determination of the terminal absorption in the DMSO by eliminating the DMSO as a solvent blank at around 200 nm, so the baicalin in DMSO and each The absorption of the product disappeared around 200 nm. The baicalin and each product have slightly different wavelengths at other main absorption peaks in two different solvents (the maximum absorption wavelength of each substance in DMSO is slightly larger than the maximum absorption wavelength of each substance in the alkali solution). value).

 The maximum absorption wavelength of baicalin and each substance measured in the lye, baicalin has three maximum absorptions at 214 nm, 275 nm and 317 nm, and all products have only two ultraviolet absorptions, and absorption disappears at 317 nm.

 The maximum absorption wavelength of baicalin and each substance measured in DMSO, baicalin has two maximum absorptions at 280 nm and 316 nm, and all products have only one ultraviolet absorption of 292 nm, and the ultraviolet absorption at 316 nm disappears.

 It can be seen that the two main absorption peaks relative to baicalin (275 nm and 317 nm in alkali, 280 nm and 316 nm in DMSO), the UV absorption peak of the complex changes significantly, one absorption peak disappears, and one absorption peak occurs. Red shift. It indicates that baicalin did react with metal ions. The reason for the red shift of ultraviolet absorption is that the formation of complexes increases the degree of delocalization of electrons in the whole molecule and the metal ions have a certain electron-withdrawing ability. The electrons of the conjugated ring system move toward the metal at the center of the plane, making electrons The excitation energy required for the transition is reduced, so the absorption peak is red-shifted.

 1-2-2 Infrared characterization of baicalin metal complexes

 Infrared results analysis of baicalin, product A (xanthine: copper sulfate or copper chloride = 2: 1, 1 : 1 or 1: 2), product B (xanthine: copper acetate = 2: 1 or 1 : 1) as follows:

The absorption peak (expansion) of C=0 is between 1850 and 1600 cm- ¹ . In the infrared spectrum of baicalin, the absorption peak (stretching vibration peak) of baicalin at position 4 was 1661.05 cm - the carbonyl absorption peak of the carboxylic acid on baicalin was 1726.71 cm" ¹ , while in the two complexes In the infrared spectrum, the two C=0 absorption peaks are broadened and merged into one peak, which is red shifted to 1624.260^, 1621.90cm" ¹ 1619.20cm- ¹ , 1625.84cm" ¹ 1620.91cm" ¹ 1623.67cm" ¹ , resulting in absorption. The reason for the peak red shift is that the lone pair of oxygen forms a coordination bond with the empty orbit of the metal ion and The metal ions delocalize the electrons, causing the electron cloud density of the carbon-oxygen double bonds to decrease, and the force constant to be small, resulting in a red shift phenomenon. Thus, the binding site of baicalin to a metal ion may be a carbonyl group at the 4-position or a carbonyl group at the carboxyl group of the glycoside.

Alcohols, phenols free bond absorption peak between 0_H 3650_3580cm- ^1, between 3400- 3200cm ^1, and the bond between 0_H 0- H bond absorption peak of the free acids 3540- 3350cm- ¹ association. The 5-position hydroxyl group of baicalin and the 4-position carbonyl group can be intramolecularly hydrogen-bonded, and the peak is located between 3400-3200 cm- ¹ . The hydroxyl group on the carboxylic acid in baicalin is free of 0-H bond, and the peak is located at 3540-3350 cm. - between ¹ . Since these 0-H absorption peaks are wide, and there are many hydroxyl groups at other positions, these 0-H absorption peaks form a large broad peak between 3600-3200 cm- ¹ , which is difficult to distinguish each 0-H. s position. In the infrared spectrum of baicalin, the large broad peak formed by these 0-H bonds has an absorption peak at 3394.45 cm- ¹ . In the IR spectrum of the complex, the complex formed baicalin, respectively, the peak position moved from 3398.45cm- ¹ 3412.63cm- 3399.95cm ^1, i.e., blue-shifted absorption peaks occurred, indicating the destruction of intramolecular hydrogen bonds, It can be seen that the hydroxyl group of the carboxyl group at the 5-position hydroxyl group or glycoside is coordinated with the metal ion.

 Thus, the hydroxy group and the hydroxyl group of the carboxyl group at the 5-position hydroxyl group, the 4-position carbonyl group or the (and) glycoside of the baicalin are coordinated with the copper ion.

 From the results of UV and IR, the UV and IR absorption of the product relative to the raw material (xanthine) changed significantly, indicating that the baicalin reacted with copper ions. The coordination site was 4 carbonyl, 5 hydroxyl or glycoside. a carbonyl group or a hydroxyl group of a carboxyl group.

 Because the UV and IR data can only identify the complex reaction of baicalin with metal ions and preliminarily determine the coordination sites where the complex reaction may occur, the exact position of the different products obtained from different ratios of different raw materials needs to be coordinated. The following methods were further identified.

1H NMR Characterization of 1-2-3 Astragalus Copper

 Baicalin 1H NMR (DMSO) data is as follows:

 δ 12.545 (s, 1H, 5-OH), 8.664 (s, 1H, 6-OH), 8.070 (d, 2H, 2', 6'-H), 7.571-7.534 (m, 3H, 3', 4 ',5'-H), 7.004 (s, 1H, 8-H), 7.000 (s, 1H,3-H), 5.289-5.499 (m, 3H, 2", 3", 4"-OH), 5.240 (d, 1H, Γ-Η), 4.060 (d, 1H, 5"-H), 3.330-3.450 (m, 3H, 2", 3", 4"-H).

 Astragalus copper (A) 1H NMR (DMSO) results and analysis are as follows:

 Due to the paramagnetism of copper ions, the peak shape of the Astragalus copper complex is broadened and the signal is weakened. However, with respect to the displacements of baicalin, the complexes have corresponding peaks, and only the peak of about 12.6 (ie 5-OH peak) ) disappears, showing that 5-OH is replaced by a H atom due to coordination with copper ions.

Due to the paramagnetic effect of copper ions, the peak shape and signal of the hydrogen spectrum of the product are difficult to judge the specific structural information very accurately. Therefore, only the hydrogen spectrum of a product (xanthine: copper chloride = 1:1) is made, but It is determined that the peak shape is widened, The weakening of the signal is an important feature of the influence of diamagnetic copper ions on the nuclear magnetic spectrum of the compound, which leads to a fundamental change in the hydrogen spectrum information of baicalin, indicating that the copper ion has been successfully complexed, and the xanthine copper (jaundice) Glycoside: Copper chloride = 1 : 1 ) Since the peak of about 12.6 (ie, the baicalin 5-OH peak) disappears, it can be preliminarily determined that the structure is coordinated with copper ions at the 5-position hydroxyl group.

1-2-4. Determination of copper content

 The copper ion content of baicalin and different copper salt complexes with different molar ratios is shown in Table 3.

Table 3 Results of copper ion content measured by reaction of baicalin with copper ions

 Baicalin: Copper chloride 10.49% 10.78% 10.91% Baicalin: Copper sulfate 10.31% 10.56% 10.76% Baicalin: Copper acetate 10.55% 11.36%

 It is preliminarily determined from the measured content of copper ions contained in each complex, baicalin and copper chloride or copper sulfate at three ratios (2:1, 1:1, 1:2) and baicalin and copper acetate The product at a ratio of 2:1 or 1:1 is one molecule of baicalin complexed with one molecule of copper ions.

 1-2-5 Physical properties

The product properties of baicalin with copper sulfate or copper chloride in different molar ratios (2:1, 1:1, 1:2) are identical, all are brownish yellow whole-sheet solids, and both are soluble in DMSO, DMF, difficult Soluble in organic solvents such as methanol, ethanol, water, ethyl acetate, etc., all dissolved in NaHC0 ₃ , Na ₂ C0 ₃ solution and dilute NaOH solution.

The product properties of baicalin and copper acetate in the molar ratio of 2:1 and 1:1 were consistent. They were all brown powdery solids, both soluble in DMSO, DMF, soluble in dilute NaOH solution, but insoluble in NaHC0 ₃ And Na ₂ C0 ₃ solution.

Baicalin with aluminum sulfate or aluminum chloride obtained a reddish brown irregular solid at a molar ratio of 2:1, a 1:1 or 1:2 molar ratio gave an orange-red irregular solid, both soluble in DMSO, DMF, It is insoluble in organic solvents such as methanol, ethanol, water and ethyl acetate, and is soluble in NaHC0 ₃ , Na ₂ C0 ₃ solution and dilute NaOH solution.

Baicalin with ferric sulfate or ferric chloride at a molar ratio of 2:1 gives a black solid, 1:1 or 1:2 molar ratio gives a black-green blocky solid, both soluble in DMSO, DMF, poorly soluble An organic solvent such as methanol, ethanol, water or ethyl acetate is dissolved in NaHC0 ₃ , Na ₂ C 0 ₃ solution and dilute NaOH solution.

1-2-6 further identification of coordination sites

According to "Natural Medicinal Chemistry"(People's Health Publishing House, Wu Lijun), it has been determined that the hydroxy acid size of each flavonoid is aligned and the solubility in lye of different strengths. The acidity of each group is as follows: carboxylic acid group (- COOH) 〉7,4′-diOH>7-OH or 4′-OH>General phenol OH>5-OH>3-OH, containing carboxyl group or both 7 and 4' The hydroxyl group is soluble in NaHC0 ₃ , only contains a guanidine or 4' hydroxy group is soluble in Na ₂ C0 ₃ , and the other hydroxyl group is insoluble in NaHC0 ₃ and Na ₂ C0 ₃ , and can only be dissolved in different concentrations of NaOH.

The product properties of baicalin with copper sulfate and copper chloride in different molar ratios (2:1, 1:1, 1:2) were identical, all were brownish yellow whole-sheet solids, and the Cu ²⁺ content was also consistent. From the results of infrared identification, due to the action of electron absorption by copper ions, the carbonyl and carbonyl groups on the carboxylic acid are red-shifted and overlapped into broad peaks, and the infrared absorption of each hydroxyl group is 3600-3200 cm ^{1 .} There is a large broad peak between them, so it is difficult to judge the coordination sites of copper ions by infrared spectrum alone. The obtained product obtained by the reaction is all dissolved in NaHC0 ₃ , indicating that the carboxyl group is present, and according to the determination of the metal ion content, the coordination ratio of baicalin to Cu ²⁺ is 1:1, and one molecule of baicalin can be combined with one. Molecular Cu ²⁺ , because the bain of the baicalin can be coordinated in two places: a carbonyl group on the carboxyl group and a hydroxyl group, a 4-position carbonyl group and a 5-position hydroxyl group, so the reaction is a molecule of baicalin with a 4-position carbonyl group and a 5-position The hydroxyl group is coordinated to a molecule of Cu ²⁺ .

The product traits of baicalin and copper acetate in the molar ratio of 2:1, 1:1 were consistent, all were brown-red powdery solids, and the Cu ²⁺ content was also consistent. The infrared results also show that the complex has a fundamental change with respect to baicalin. The change sites are also carbonyl and hydroxyl groups, but as with the above results, the absorption peaks of the same groups overlap, making it difficult to determine the specific coordination sites simply by infrared spectrum. The resulting reaction product was dissolved in NaOH, and insoluble NaHC0 ₃ and Na ₂ C0 ₃ solution, indicating the presence on the ring -OH, -COOH hydrogen glycosides and no longer exists. According to the determination of metal ion content, the coordination ratio of baicalin to Cu ²⁺ in the product is 1:1, one molecule of baicalin can be combined with one molecule of Cu ²⁺ , and the coordinating group which can occur due to baicalin is Two: a carboxyl group, a 4-position carbonyl group and a 5-position hydroxyl group, which are changed by the absorption of the infrared carbonyl group and the hydrogen of the -COOH of the glycoside is absent. It is known that one molecule of baicalin occurs with a carbonyl group at the carboxyl group and a hydroxyl group with a molecule of Cu ²⁺ . Coordination.

Characterization of 1-2-7 Astragalus Copper Complex by LC-MS

 The xanthine copper complex A and the xanthine copper complex B were analyzed by mass spectrometry. The molecular ion peaks obtained in the mass spectrum were:

 Complexes Molecular ion peaks Astragalus copper A (products obtained from different contents of baicalin and copper sulfate or copper chloride) 590

 Astragalus copper B (the product obtained from the combination of baicalin and copper acetate in a ratio of 2:1 or 1:1) 568

 In summary, the structure of each substance is estimated to be as follows:

1. The structure of baicalin reacted with different proportions of copper chloride or copper sulfate to obtain the following structure: (The relative molecular mass of the structure is 589, and the proportion of copper is 10.70%)

 III

Mass spectrometry: The peak position of the molecular ion is shown to be 590. Since it is in the (molecular + H ⁺ ) mode, the actual molecular weight is 589, which is in complete agreement with the molecular weight of the above structure. When bombarding molecular ions, it is shown as 588 due to ionization. Fragment ion peak 570 has the highest intensity, and 588-570=18 is obviously caused by the loss of a part of the molecular ion peak (the water molecule should be the coordination water on the metal copper, so it is lost first). The intensity of the fragment peak 475 is also large, according to 588-18-63-32=475, probably because the molecular ion peak loses one molecule of coordination water (18), one molecule of coordinated methanol (32), and one molecule of copper ions ( 63). The peak intensity of the fragment peak 361.9 is second only to the peak intensity of 570, 361.9 = 588-18-176-32. It can be seen that this is another way of cleavage of the molecular ion peak, that is, the molecular ion loses one molecule of coordination water (18). One molecule of coordinated methanol (32) and one molecule of glycoside (176). Fragment peak 330.8=588-18-176-32-31, it can be seen that this cleavage mode continues to lose one molecule of coordination water (18), one molecule of coordinated methanol (32) and one molecule of glycoside (176). Loss of one molecule of -OCH ₃ group bound to copper ions. It can be seen that the structure consists of one molecule of baicalin combined with one molecule of copper ions, and another molecule of water and two molecules of methanol are respectively coordinated with copper ions.

 2. The structure of baicalin and copper acetate (2:1, 1 : 1) is estimated as follows: (The relative molecular mass of the structure is 567, and the proportion of copper is 11.11%)

 IV

Mass spectrometry: The molecular ion peak was 568. Since it was (Molecular + H ⁺ ) mode, the actual molecular weight was 567, which was in complete agreement with the molecular weight of the above structure. Example 2 Pharmacodynamic experiment In vitro bacteriostatic test, the results of bacteriostatic experiments are shown in Table 4.

Table 4 Antibacterial experiment results (MIC value)

Baicalin 2.5mginl 2.5mg ^/ ml Baicalin: Copper chloride or copper sulfate * 0.5mginl 0.25mginl Baicalin: Copper acetate (2:1 or 1:1) O^mginl O mginl Copper chloride 2 mginl 1 mginl

 Copper sulfate 2 mginl 1 mginl

 Copper acetate 2 mginl 1 mginl

 * Since the products obtained from baicalin and copper chloride and copper sulfate at the three ratios (2:1, 1:1, 1:2) are the same product, the antibacterial effect is the same, so only one data is listed.

 **Because the product obtained from the baicalin and copper acetate at the ratio of 2:1 and 1:1 is the same product, the antibacterial effect is also the same, so only one data is listed.

 Results: It can be seen from the bacteriostatic results that the antibacterial effect of each complex is significantly enhanced compared with baicalin. The MIC of the complex obtained by baicalin with copper sulfate (or copper chloride) against Staphylococcus aureus and Escherichia coli is 1/5, 1/10 of baicalin, the MIC of baicalin and copper acetate (2:1 or 1:1) for Staphylococcus aureus and Escherichia coli is 1/10, 1/ of baicalin, respectively. 5.

 Cai Chuanying's published patent (Publication No.: CN 1462619A) Invented a zinc baicalin (structured as a molecule of baicalin with a molecular weight of zinc ion) pharmaceutical preparation, with good antibacterial effect, MIC against Staphylococcus aureus 0.63 mg1⁄2l, the MIC for Escherichia coli is 5.0 mg1⁄2l; Wang Yutian et al. (Publication No.: CN 1634952A) Invented a zinc baicalin complex (structure is two molecules of baicalin with carboxyl group, 4-position carbonyl group and 5 The hydroxyl group is complexed with three molecules of zinc ion), the MIC for S. aureus is 1 mg1⁄2l, and the MIC for E. coli is >1 mg1⁄2l. However, the inhibitory effect of the two kinds of xanthine copper complexes of the invention on S. aureus is 1.26-4 times that of the baicaloside complex invented by the above patent, and the inhibitory effect on Escherichia coli is the combination of the baicalin invented by the above patent. The two yellow bismuth copper complexes invented in this patent show stronger antibacterial activity. Example 3. Antibacterial test of the product of the invention against different drug resistant bacteria

The minimum inhibitory concentration (MIC) of six compounds against resistant Escherichia coli, Salmonella and Staphylococcus aureus. 1 Materials and reagents

 1.1 Compound

 1#-Astragalus (flavonoid extract), 2# - Astragalus copper complex A, 3# - Astragalus copper complex B, 4# - Astragalus copper mixture (1: 1), 5# - copper sulfate, 6# - Astragalus zinc (prepared and verified by the method of Example 1, its structure is similar to Astragalus copper complex A)

1.2 Test strain

 Escherichia coli CVCC1498 (C83543) was isolated from diarrhea piglets in a farm in Guangxi in 1980. The blood type was 0141:K+. Resistant to tetracycline, penicillin, sulfonamides and other drugs. Porcine Escherichia coli CVCC1515 (C83646 P1038), isolated from the digestive tract of diarrhea piglets, serotype 0149, multi-drug resistant (ampicillin-resistant, sulfamethoxazole, co-trimoxazole, tetracycline, doxycycline, ceftiofur, etc. drug). Salmonella typhimurium CVCC 533 (C79-13): It was isolated from a chicken farm in Maliantun, Xijiao, Beijing, and isolated from the heart of the diseased chicken. The serotype was 9,121,123. Resistance to tetracycline, penicillin, sulfonamides. Recorded in: Ma Weiming et al., Antibacterial effect of porcine intestinal antibacterial peptides, Chinese Journal of Veterinary Medicine, 2005, first issue. Salmonella henensis CVCC1793 CSal.5, 15616y ): Isolated in the digestive tract of chicken, strong pathogenic, resistant to amyloid, polymyxin. S. aureus S. aureus 546 (C56023) In 1989, it was isolated from the joints of dead pigs in a pig farm in Haidian, Beijing, and was resistant to ampicillin and ciprofloxacin. Recorded in: Fu Yaxiang, etc., chemical composition analysis and antibacterial effect research, "Advances in Animal Medicine", 2008, 29 (9): 45~49. Staphylococcus aureus CVCC 2086 (ATCC, 6538): Causes suppuration Sexual disorders, multi-drug resistance (penicillin, ampicillin, ampicillin/clavulanic acid, clindamycin, erythromycin, sulfisoxazole, sulfamethoxazole, ceftiofur, cefoxitin, ofloxacin Magnitude). The above resistant strains were purchased from the China Veterinary Microorganisms Collection (CVCC). The numbering is shown in the CVCC Veterinary Microbial Species Sharing Database.

Http:〃222.35.47.100:8018/cvcc/cvcc search/search fet.isp, which is guaranteed to be issued to the public for verification testing within 20 years from the date of filing.

1.3 Instruments and reagents

C0 ₂ incubator; balance (sensing O.OOOlg); 96-well plate; pipette (20 (^L and ΙΟΟΟμυ; turbidity meter; sterile filter (ρ=0.22μηι); MIC bacterial dilution (normal saline) ); MH broth; NaHC0 ₃ solution. 2 methods

 2.1 Configuration of various compound solutions

2.1.1 NaHC0 ₃ solution

Weigh accurately NaHC0 ₃ 0.03g, was dissolved in sterile deionized water, dissolved set to 100ml, that is, 0.03% NaHCO ₃ solution.

2.1.2 Compound solution: Weigh 40mg of compound and dissolve it in 10 ml of the above NaHC0 ₃ solution to a final concentration of 4mg/ml.

( _1# and 4# slightly soluble, sonicated to full dissolution), compound No. 5 was dissolved directly in 10 ml of sterile deionized water.

2.2 Resuscitation of the strain Escherichia coli 1498 (2010.3.16 freeze-dried), Salmonella 533 (2005.4.6 freeze-dried) and Staphylococcus aureus 546 (2002.1.16 lyophilized) were reconstituted with physiological saline and inoculated on a nutrient agar plate, large intestine The glycerol cryopreservation solution of Bacillus CVCC1515, Salmonella CVCC1793 and Staphylococcus aureus CVCC 2086 was directly inoculated on a nutrient agar plate and placed in a 37 ° C incubator for 20 h. After the cultivation, a single colony was picked and inoculated with a nutrient agar plate for one generation.

2.3 Determination of MIC

2.3.1 Dilution of the compound: 0.3ml of 6 compounds and 4% NaHC0 ₃ solution were added to the first column of wells of 96-well plate (8*12) (each compound corresponds to a row of holes, occupying 7 rows of holes, The last row of wells was used as a growth control and blank control), and 0.15 ml of MH broth was added to each of the 7 rows of wells. Pipette 0.15ml of the compound solution from the first column of holes into the second column of wells, do the dilution, mix by pipetting, and then transfer the second column of 0.15ml solution to the third column of holes, and so on, until the 8th column. After the wells have been diluted, remove 0.15 ml of the solution and discard. The final compound concentration per well is as follows:

2.3.2 Antibacterial test Two to two colonies of different test strains grown on nutrient agar plates were picked and dissolved in MIC bacterial dilution (physiological saline) to adjust the turbidity of the bacterial suspension to 0.5 Mei units. Pipette 0.3 ml of the bacterial suspension into 30 ml of MH broth, make a 100-fold dilution, and then dilute the bacteria. The solution was added to a 96-well plate containing various concentrations of compound at 0.15 ml per well. Two of the eight wells in the last row (the first 4 wells and the last 4 wells) were each added with 0.15 ml of MH broth and bacterial dilution as a blank control and growth control. The 96-well plate was then placed in a 37 ° C incubator.

2.3.3 Read the antibacterial test results after 20h

3 results

3.1 Antibacterial test results (MIC values) Combining two parallel tests, the bacteriostatic test results (MIC) are shown in Table 6 below:

3.2 Antibacterial test results for ampicillin (MIC value)

 Selection of the drug-resistant strains of S. aureus: a combination of two parallel test results, the bacteriostatic test results (MIC value mg / ml)

注： 抗菌药 +2号指的是在稀释好的细菌中每 10ml加入 0.1ml的 黄岑甙铜化合物 B， 混 匀后再加入系列稀释好的抗菌药中， 测定 MIC。

Note: Antibacterial + No. 2 refers to the addition of 0.1 ml of Astragalus Copper Compound B per 10 ml of diluted bacteria. After mixing, add a series of diluted antibacterial drugs to determine the MIC.

3.3 Analysis of results Different test products have different MICs for different bacteria. 2# and 3# compounds have the strongest antibacterial effect on Escherichia coli, MIC is 1 mg/ml, followed by 4# and 5# compounds, MIC is 2 Mg/ml, 1# and 6# are >2 mg/ml. The antibacterial effects of the compounds on Salmonella were similar, with 2#, 3#, 4# and 6# being slightly stronger at 0.5 mg/ml, and 1# and 5# being 1 mg/ml and 2 mg/ml. The antibacterial effect of the compound against Staphylococcus aureus is stronger than that of Escherichia coli and Salmonella. The 2# and 3# compounds have the strongest antibacterial effect against Staphylococcus aureus. The MIC is 0.125-0.25 mg/ml, 4# and 6# compounds. The MIC is 0.25-0.5 mg/ml, and the MIC of the 1# and 5# compounds is 1 mg/ml. The MIC of NaHC0 ₃ dilution for all three bacteria was >2, indicating that the NaHC0 ₃ dilution did not affect the bacteriostatic effect.

 The strains selected in this experiment all have different degrees of drug resistance. However, the bacteriostatic effect of the tested products did not show the difference in bacteriostatic effect caused by the different degree of resistance of the bacteria, and the same product had the same antibacterial effect on different drug-resistant strains. As shown in Table 7, the compounds of the present invention also reduce the resistance of S. aureus to ampicillin. 4 Conclusion

4.1 The inhibitory effects of the six test products on the three tested bacteria were different. In general, the inhibition of Staphylococcus aureus was the strongest, followed by Salmonella, and the inhibition of Escherichia coli was the weakest.

4.2 Astragalus copper complex has the best antibacterial effect, superior to Astragalus copper mixture and copper sulfate, which is significantly larger than Astragalus and Astragalus. The MIC for E. coli is 1 mg/ml, which is 1/2 of the mixture of astragalus and copper sulfate, and is less than 1/2 of that of xanthine and xanthine. The MIC for Staphylococcus aureus is 0.125-0.25 mg/ml, which is 1/2 of the mixture of baicalin and xanthine, and is less than 1/4 of baicalin and copper sulfate. There was no difference in the MIC of different bacteria for Astragalus copper 1 and Astragalus copper.

 4.3 The antibacterial effect of this test product is not affected by the degree of bacterial resistance. Example 4. Antitumor effect of Astragalus copper of the present invention

Lung cancer A549 cells and HepG2 cells in logarithmic growth phase were diluted with RPMI-1640 medium containing 10% by volume of fetal bovine serum to a concentration of lx l0 ⁴ /mL, and inoculated into 96 wells. Plate, per well ΙΟΟμΙ^, incubate in the incubator at 37 ° C, C0 ₂ volume fraction of 5% for 24 hours, discard the medium, add fresh medium ΙΟΟμΙ ^, then add different concentrations (20 , 40, 80, 160, 320μΜ) of baicalin or xanthine copper solution, each well ΙΟΟμ each set of 5 duplicate wells, and set up the solvent control group, continue to culture under the same conditions for 48 hours, then add each well Incubate for 4 hours, discard the supernatant, and add DMSO to shake. The absorbance was measured at 490 nm using a microplate reader, and the average of 5 replicate wells was taken to calculate the half effective inhibitory concentration (IC _5Q ) of each compound against different tumor cells. The results are shown in Table 6.

 Table 6 Antitumor activity of baicalin and its metal complexes

 As can be seen from Table 6, the inhibitory activities of the two kinds of xanthine copper prepared in Example 1 on lung cancer A549 cells and liver cancer HepG2 cells were significantly enhanced compared with baicalin. There is no literature on the anti-tumor effect of baicalin metal complexes.

 Based on the above experimental results, the two kinds of xanthine copper prepared in Example 1 can be used for the preparation of antibacterial drugs and antitumor drugs. Example 5 Preparation and use of baicalein metal complex

 The xanthine copper compound prepared in Example 1 was used as a starting material, and the baicalein copper complex was prepared by the following process:

 Astragalus copper is dissolved in 10% hydrochloric acid, and hydrolysis reaction occurs. The hydrolyzate is separated by column chromatography to obtain baicalein copper.

 The process is as follows:

Baicalin

 It was verified by the functional experiments of Examples 3 and 4 that there was no difference between the acute toxicity and pharmacological experiments prepared in the same manner as in Example 1.

Claims

Claim 1. A yellow copper complex with a structure as shown in formula (III):

 III  The method for preparing a xanthine copper complex according to claim 1, characterized in that: baicalin, copper sulfate or copper chloride is dissolved in a non-aqueous solution, respectively, and the molar ratio is 2:1, 1: The mixed reaction of 1 or 1:2 gives a brownish yellow precipitate, which is filtered, washed several times with ethanol, and dried. The nonaqueous solution is one or more of methanol, ethanol, ethyl acetate and petroleum ether.  The method for preparing a xanthine copper complex according to claim 1, wherein the reaction conditions are: a reaction temperature of 30-80 ° C and a reaction time of 2-3 hours.  4, Astragalus copper with its structure as shown in formula (IV):

 IV  The method for preparing a xanthine copper complex according to claim 4, wherein: the baicalin and the copper acetate are respectively dissolved in a non-aqueous solution, and the molar ratio of the two is 2:1 or 1:1. The precipitate is red, filtered, washed several times with ethanol, and dried. The non-aqueous solution is one or more of methanol, ethanol, ethyl acetate and petroleum ether. 6. The method for preparing a xanthine copper complex according to claim 5, wherein the reaction conditions are: The reaction temperature is 30-80 ° C, and the reaction time is 2-3 hours.  7. Use of a xanthine copper complex according to claim 1 or 4 as a servo additive.  The use of the xanthine copper complex according to claim 1 or 4 for the preparation of an antibacterial or antitumor drug.  9. The use according to claim 8, wherein the antibacterial drug refers to a drug resistant to a drug, the drug resistant bacteria refers to a drug resistant pig-derived Escherichia coli; a drug resistant Salmonella typhimurium or a drug resistant Staphylococcus aureus .  10. The use according to claim 8, wherein the anti-tumor drug refers to a drug against lung cancer or anti-liver cancer.  An antibacterial agent, characterized in that the antibacterial active substance comprises the xanthine copper complex according to claim 1 or 4.  12. The antibacterial agent according to claim 11, further comprising a traditional Chinese medicine, a western medicine or other harmless chemical composition beneficial to the antibacterial compound which is compatible with the xanthine copper complex.  13. The antibacterial agent according to claim 11, which further comprises a pharmaceutically acceptable auxiliary ingredient. The antibacterial agent according to any one of claims 11 to 13, which is in the form of a capsule, an injection, a tablet, a granule, an oral solution, an ointment, a powder or a suspension.  An antitumor drug, characterized in that the antitumor active substance comprises the xanthine copper complex according to claim 1 or 4.  16. The antitumor agent according to claim 15, further comprising a traditional Chinese medicine, a western medicine or a harmless chemical composition thereof which is beneficial for antitumor, which is compatible with the xanthine copper complex.  The antitumor agent according to claim 15, which further comprises a pharmaceutically acceptable auxiliary component.  The antitumor agent according to any one of claims 15 to 17, which is in the form of a capsule, an injection, a tablet, a granule, an oral solution, an ointment, a powder or a suspension.  19. The baicalein metal complex is a product obtained by dissolving the baicalin metal complex according to claim 1 in a 10% hydrochloric acid solution for hydrolysis and hydrolyzing the product by column chromatography.