Black ginger extract and application thereof in improving bioavailability of berberine hydrochloride sample

Abstract

The invention relates to the technical field of biology, and discloses a black ginger extract and application thereof in improving the bioavailability of a berberine hydrochloride sample. Specifically, the research finds that the combined application of the black ginger extract and berberine hydrochloride can significantly improve the bioavailability of the berberine hydrochloride, and enhance the strength and duration of the pharmacological action of the berberine hydrochloride, and the result provides a scientific basis for the development of an innovative formula of the combined application, and shows the important value of the berberine hydrochloride in clinical application.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to a black ginger extract and an application thereof in improving the bioavailability of a berberine hydrochloride sample. Background Art

[0002] Berberine hydrochloride, as a compound with significant biological activity in the field of medicine and health care products, is widely used in a variety of treatments and health care products. However, its bioavailability is often affected by a variety of complex factors, such as pH value, intestinal environment and interaction with other drugs, resulting in unsatisfactory absorption and utilization in the human body. In order to effectively overcome this problem, scientific researchers and medical experts continue to explore and research, aiming to find effective strategies that can significantly improve the bioavailability of berberine hydrochloride.

[0003] In current research, a variety of technical means have been proposed and tried to enhance the bioavailability of berberine, including but not limited to modification of chemical structure, improvement of formulation technology, and combined application with active ingredients with synergistic effects. Chemical modification is mainly aimed at optimizing the molecular structure of berberine hydrochloride, aiming to enhance its stability and absorption efficiency in the body. Improvements in formulation technology focus on the development of new drug formulations, such as the use of nanotechnology and microemulsification technology to improve the solubility and bioavailability of drugs. In addition, combined application with active ingredients such as vitamin C and specific plant extracts has also been shown to enhance the absorption and efficacy of berberine. For example, in the study of Mishra R and Dhole S, berberine hydrochloride solid dispersions were prepared by using different proportions of hydrophilic carrier glycerol 44 / 14 and glycerol 50 / 13 to improve its bioavailability (see Mishra R, Dhole S. Lipid-based floating multiparticulate delivery system for bioavailability enhancement of berberine hydrochloride [J]. J Appl Pharm Sci, 2019, 9: 36-47.); Li et al. developed an oil-in-water nanoemulsion system for berberine hydrochloride to improve its stability and oral bioavailability (see Li YJ, Hu XB, Lu XL, et al. Nanoemulsion-based delivery system for enhanced oral bioavailability and Caco-2 cell monolayers permeability of berberine hydrochloride [J]. Drug Deliv, 2017, 24: 1868-1873.). Although the above methods can improve the bioavailability of berberine to a certain extent, they still face challenges and limitations such as high cost, complex production process or insufficient effect.

[0004] Therefore, developing a novel, efficient and economical method to improve the bioavailability of berberine hydrochloride not only has important scientific value, but also has vital significance for promoting the market application and popularization of related pharmaceutical products. Summary of the invention

[0005] As a traditional herbal medicine, black ginger has multiple pharmacological activities and has gradually attracted attention in the field of medicine and health products in recent years. After in-depth research, the inventors found that black ginger extract can significantly improve the bioavailability of berberine hydrochloride, thereby providing new possibilities for its application in the field of medicine and health products. 5,7-Dimethoxyflavanone in black ginger is a compound with unique pharmacological activity, which can produce synergistic effects with berberine hydrochloride and effectively promote its absorption and utilization. In addition, other active ingredients in black ginger extract may also have a positive effect on the intestinal microenvironment, further promoting the absorption and transport of berberine hydrochloride. Through the method disclosed in the present invention, the bioavailability of berberine hydrochloride in the human body can be significantly improved, thereby improving its therapeutic effect, reducing the dosage of medication, reducing the economic burden of patients, and reducing the potential risk of side effects. Therefore, the present invention provides a new, efficient and economical solution for the application of berberine hydrochloride in the field of medicine and health products.

[0006] Berberine has gastrointestinal adverse reactions after taking, such as nausea, vomiting, diarrhea, etc., and these adverse reactions limit its clinical application to a certain extent. The present invention, by introducing black ginger extract, can not only effectively improve the bioavailability of berberine hydrochloride, but also reduce or alleviate its gastrointestinal adverse reactions by stopping vomiting and improving the intestinal microenvironment, thereby further improving the patient's medication experience and therapeutic effect. In addition, black ginger, as a natural herbal medicine, is rich in resources and relatively cheap, and the preparation process of its extract is also relatively simple, which makes the present invention have high economy and feasibility in practical application.

[0007] In view of this, the present invention proposes a black ginger extract and its application in improving the bioavailability of berberine hydrochloride samples, aiming to optimize the bioavailability characteristics of berberine hydrochloride by combining the pharmacological activity of the black ginger extract. Specifically, the present invention provides a method for preparing a black ginger extract, and a technical solution for combining the extract with berberine hydrochloride to improve its bioavailability.

[0008] The invention provides a black ginger extract and application thereof in improving the bioavailability of a berberine hydrochloride sample.

[0009] Preferably, the preparation method of the black ginger extract comprises:

[0010] The black ginger is subjected to alcohol extraction and then filtered to obtain a primary extract and an extraction residue, and the extraction residue is subjected to alcohol extraction and filtration again to obtain a secondary extract; the primary extract and the secondary extract are mixed and concentrated to 1 / 3-1 / 5 of the original volume to obtain a concentrated solution; the concentrated solution is adsorbed by a resin to obtain a supernatant, and the supernatant is analyzed by a column using ethanol to obtain a black ginger extract; the black ginger extract is dried to obtain the black ginger extract.

[0011] Preferably, 50%-60% ethanol solution is used for alcohol extraction.

[0012] Preferably, the preparation method of berberine hydrochloride comprises:

[0013] After the piperidine is dissolved in the solvent I, it enters into a condensation process to obtain a condensation product;

[0014] The condensed product is dissolved in solvent II, a catalyst is added to carry out a hydrogenation reaction, and after the reaction is completed, the catalyst is removed by filtering to obtain a hydrogenated product;

[0015] The hydrogenation product is mixed with hydrochloric acid to generate a salt compound, and then a condensation reagent is added to carry out a condensation reaction to obtain a condensation product;

[0016] The condensation product is added to a solvent containing copper salt to carry out a cyclization reaction. During the reaction, a chelating agent is added to remove copper ions. After the cyclization is completed, a crude product is obtained;

[0017] Dissolve the crude product in hot water, maintain the temperature at 50±5°C and stir; remove insoluble impurities through a filter press to obtain a clear solution;

[0018] The clear solution is dried to obtain the berberine hydrochloride.

[0019] Preferably, the solvent I is anhydrous ethanol; the solvent II is methanol; and the condensation reagent is phosphorus trichloride.

[0020] Preferably, the condensation process comprises:

[0021] The solution of piperonyl ethylamine dissolved in solvent I was cooled to 5° C. under stirring, and a condensation reagent was added dropwise to carry out a condensation reaction, and the reaction time was maintained for at least 1 hour. After the condensation was completed, nitrogen protection was used, the temperature was raised to room temperature, and the condensation product was collected to obtain the condensation product.

[0022] Preferably, the condensation agent is formamide.

[0023] The present invention also provides a product for improving the bioavailability of a berberine hydrochloride sample, wherein the product comprises a black ginger extract.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] When black ginger extract is used in combination with berberine hydrochloride, the rich bioactive ingredients in black ginger extract can promote the absorption and distribution of berberine hydrochloride and optimize its pharmacokinetic behavior in the body. Specifically, black ginger extract can significantly increase the plasma peak concentration (Cmax) and total drug exposure (AUC) of berberine hydrochloride and prolong its half-life (T 1 / 2 ). This combined use not only improves the bioavailability of berberine hydrochloride, but also significantly enhances the intensity and duration of its pharmacological effects, providing a scientific basis for the development of innovative formulas for combined use, and has important clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Various other advantages and benefits will become apparent to those of ordinary skill in the art by reading the detailed description of the preferred embodiments below. The accompanying drawings are only for the purpose of illustrating the preferred embodiments and are not to be considered as limiting the present invention. Moreover, the same reference symbols are used throughout the accompanying drawings to represent the same components. In the accompanying drawings:

[0027] Figure 1 is the standard curve;

[0028] Figure 2 is the blood concentration-time curve of berberine hydrochloride;

[0029] Figure 3 This is the blood concentration-time curve of berberine hydrochloride + black ginger extract;

[0030] Figure 4 is the berberine blood concentration-time curve;

[0031] Figure 5 This is a pharmacokinetic correlation diagram. DETAILED DESCRIPTION

[0032] Exemplary embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. Although exemplary embodiments of the present disclosure are shown in the accompanying drawings, it should be understood that the present disclosure can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided in order to enable a more thorough understanding of the present disclosure and to fully convey the scope of the present disclosure to those skilled in the art. It should be noted that, in the absence of conflict, the embodiments of the present invention and the features described in the embodiments can be combined with each other. The present invention will be described in detail below with reference to the accompanying drawings and in combination with the embodiments.

[0033] Example 1

[0034] 1. Specific implementation process for preparing berberine hydrochloride

[0035] 1. Preparation of starting materials

[0036] Weigh piperidine (purity ≥ 99%) and dissolve it in anhydrous ethanol.

[0037] The solution was transferred to the reactor, and the temperature in the reactor was controlled at 25±2°C, ready to enter the condensation treatment step.

[0038] 2. Condensation treatment

[0039] Under stirring, the solution was slowly cooled to 5°C.

[0040] The condensation reaction was carried out by dropwise addition of a condensation reagent (formamide), and the reaction time was maintained for 1 hour.

[0041] After the condensation was completed, nitrogen was used for protection, the temperature was slowly raised to room temperature, and the condensed product was collected.

[0042] 3. Hydrogenation reaction

[0043] The condensed product was dissolved in an appropriate amount of methanol, and palladium carbon (Pd / C, 5%) catalyst was added.

[0044] Under hydrogen pressure (0.3 MPa), the reaction temperature was maintained at 20-40°C and the hydrogenation reaction was carried out for 2 hours.

[0045] After the reaction is completed, the palladium carbon catalyst is removed by filtration to obtain a transparent liquid, i.e., the hydrogenation product.

[0046] 4. Salt-forming step

[0047] Under stirring conditions, the hydrogenation product is mixed with an appropriate amount of hydrochloric acid (concentration 10%) to generate a salt compound.

[0048] The reaction temperature was controlled between room temperature and 50°C, and stirring was continued for 0.5-1 hour.

[0049] The reaction solution was cooled and filtered to collect the precipitated salt compound.

[0050] 5. Preparation of condensation products

[0051] The salt-formed product is transferred to another reaction vessel, and appropriate amounts of condensation reagent (phosphorus trichloride) and solvent (ether) are added.

[0052] The mixture was heated to 60-80°C under reflux conditions and maintained for 3-4 hours to form a condensation product.

[0053] After the reaction is completed, the solution is cooled to precipitate a solid condensation product which is recovered by suction filtration.

[0054] 6. Cyclization and copper removal

[0055] The condensation product is added to a solvent containing a small amount of copper salt (copper nitrate) and heated to 80-120° C. for cyclization reaction.

[0056] During the reaction, a chelating agent (ethylenediaminetetraacetic acid, EDTA) was added to remove copper ions.

[0057] After the cyclization is complete, the crude product is isolated using distillation or distillation under reduced pressure.

[0058] 7. Dissolution and filtration

[0059] The crude product was dissolved in hot water, maintained at 55±2°C, and stirred for 30 minutes.

[0060] The insoluble impurities are removed by filter press equipment to obtain a clear solution.

[0061] 8. Drying and final product preparation

[0062] The clear solution was transferred to a desiccator and the water was removed by vacuum drying.

[0063] After drying, a pure berberine hydrochloride product (purity ≥ 99%) is obtained.

[0064] 9. Product packaging and storage

[0065] The dried berberine hydrochloride is packaged in moisture-proof and light-proof sealed containers.

[0066] Store in a cool, dry environment to ensure product quality.

[0067] This process involves multiple key steps, and the specific experimental conditions should be optimized according to actual needs to improve the yield and purity of the product.

[0068] 2. Specific implementation process for preparing black ginger extract

[0069] 1. Raw material preparation stage

[0070] Raw material preparation: Select high-quality black ginger raw material and wash it to remove surface dirt and impurities.

[0071] Crushing process: Use a crusher to crush the black ginger into small particles (the particle size is controlled at 0.5-1cm) to increase the extraction efficiency.

[0072] Storage: Temporarily store the crushed black ginger particles in a clean container for later use.

[0073] 2. First extraction stage

[0074] Solvent preparation: Prepare 55% ethanol solution and ensure the correct solvent ratio.

[0075] Extraction equipment: Add the crushed black ginger particles into the extraction tank, add 55% ethanol, and control the amount added according to the solid-liquid ratio of 1:9 (black ginger: ethanol).

[0076] Extraction conditions:

[0077] Temperature: 70±2℃.

[0078] Duration: 1 hour.

[0079] Stirring: Stir throughout the process to ensure even extraction.

[0080] Extraction liquid collection: After the extraction is completed, the extraction liquid is collected through the drain port.

[0081] 3. First filtration stage

[0082] Filtration equipment: Use filter cloth or vacuum filter.

[0083] Operation: Filter the first extract to remove insoluble impurities.

[0084] Collection: Get the clarified first extract and keep the filtered extract residue for later use.

[0085] 4. Secondary extraction stage

[0086] Repeat extraction: Place the extraction residue in the extraction tank again, add fresh 55% ethanol solution, and extract at a solid-liquid ratio of 1:8.

[0087] Extraction conditions:

[0088] Temperature: 70±2℃.

[0089] Duration: 1.5 hours.

[0090] Stirring: Continue stirring to increase extraction efficiency.

[0091] Secondary extract collection: Collect the extract through the drain port.

[0092] 5. Secondary filtration stage

[0093] Operation: Filter the secondary extract to remove insoluble impurities.

[0094] Collection: Obtain a clarified secondary extract.

[0095] 6. Concentration stage

[0096] Mixed extract: Combine the first extract and the secondary extract and transfer them to the concentration equipment. Concentration conditions:

[0097] Temperature: 55±2℃ (under reduced pressure to avoid component damage).

[0098] Duration: 2 hours.

[0099] Operation: Concentrate to 1 / 3 of the original volume to obtain a concentrated solution.

[0100] 7. Ethanol recovery stage

[0101] Recovery equipment: Use alcohol recovery device.

[0102] Operation: Recover the ethanol in the concentrate, and the recovered ethanol can be recycled.

[0103] 8. Extraction slag treatment stage

[0104] Treatment: The extraction residue left after concentration is regarded as waste and can be used for composting or other purposes.

[0105] 9. Resin adsorption stage

[0106] Resin selection: Use macroporous adsorption resin (D101 resin).

[0107] Adsorption conditions:

[0108] The concentrate was passed through a resin column (the flow rate was controlled at 2-3 mL / min·g).

[0109] After adsorption is completed, impurities are washed away with a small amount of pure water.

[0110] Collection: The liquid after adsorption is the supernatant.

[0111] 10. Supernatant treatment stage

[0112] Filtration: Filter the supernatant using plate and frame to remove suspended particles.

[0113] Precipitation: Let the supernatant stand and add an acidic precipitant (acetic acid) to promote the precipitation of the precipitate. Discard the precipitate: Discard the precipitate after filtering.

[0114] 11. Ethanol analysis stage

[0115] Preparation of analytical solution: Prepare 75% ethanol solution.

[0116] Operation: Use analytical solution to pass through the resin column to analyze the target components.

[0117] Collection: Collect the analytical solution and filter to remove impurities.

[0118] 12. Alcohol recovery stage

[0119] Recovery equipment: recover ethanol from the analytical solution under reduced pressure conditions.

[0120] Operation: Concentrate to a certain volume to provide high concentration solution for subsequent processes.

[0121] 13. High temperature sterilization stage

[0122] Sterilization equipment: transfer the analytical solution into the sterilization tank.

[0123] Sterilization conditions:

[0124] Temperature: 121℃.

[0125] Duration: 20 minutes.

[0126] Operation: Cool to room temperature after sterilization.

[0127] 14. Spray drying stage

[0128] Drying equipment: Send the sterilized liquid into the spray dryer.

[0129] Drying conditions:

[0130] Inlet air temperature: 190-200℃.

[0131] Air outlet temperature: 90-100℃.

[0132] Collection: Obtain black ginger extract powder.

[0133] 15. Mixing stage

[0134] Mixing equipment: Use a mixer.

[0135] Operation: Mix the dried powder evenly to ensure product consistency.

[0136] 16. Packaging and Warehousing Stage Packaging: Pack the mixed powder into aluminum foil bags or sealed containers and label them. Warehousing: Store the finished products in a dry, cool warehouse.

[0137] 3. Berberine Bioavailability Experiment

[0138] 1. Experimental animals and feeding conditions

[0139] Breed: SD rat;

[0140] Level: SPF level;

[0141] The planned number of animals to be purchased, their gender and weight range: 40, male, 240-260g;

[0142] The actual number of animals used, their gender and weight range at the start of the experiment: 40 animals, male, 240-260 g.

[0143] Source: Guangzhou Ruige Biotechnology Co., Ltd.

[0144] Identification method: Write the number on the tail with a marker pen. Use the tail number and the cage number to identify.

[0145] Animal welfare: The contents and procedures of animal experiments involved in this study will comply with the relevant laws and regulations on the use and management of experimental animals and the relevant regulations of the institution's Experimental Animal Ethics Committee to ensure the welfare of experimental animals.

[0146] Euthanasia: Death by carbon dioxide; the body is temporarily stored in a body freezer, awaiting unified pollution-free disposal by this laboratory.

[0147] Quarantine and observation: The purchased rats were quarantined for three days and observed once a day. Unhealthy rats were immediately eliminated and healthy rats were selected for experiments.

[0148] Feeding and management: The animals were kept in the animal room of Nanxin Instrument (Guangzhou) Manufacturing Co., Ltd. Animal feeding conditions: 5 animals / box, group-raising; feeding temperature and humidity: 20-26℃, 40-70%, 12h:12h day and night lighting; the feeding room conditions were always kept stable to ensure the reliability of the test results. Free access to food and water, feed and drinking water were provided by Nanxin Instrument (Guangzhou) Manufacturing Co., Ltd.

[0149] 2. Dosage design and grouping

[0150] Dosage design: The dose of berberine hydrochloride is 200 mg / kg body weight, and the dose of berberine + black ginger extract is 250 mg / kg body weight (200 mg / kg berberine + 50 mg / kg black ginger extract).

[0151] Grouping: After passing the animal quarantine, rats were randomly divided into berberine group and berberine + black ginger extract group according to body weight, with 10 rats in each group. See Table 1.

[0152] Table 1

[0153]

[0154]

[0155] 3. Experimental methods

[0156] (1) Animal gavage collection

[0157] This study used 10% PEG400 saline solution for preparation, and the specific operation was to dissolve 10 mL PEG400 in 90 mL saline.

[0158] The preparation method of berberine intragastric solution is as follows: take 800 mg of berberine hydrochloride, add an appropriate amount of 10% PEG400 saline solution and mix evenly, then add to a total volume of 40 mL to prepare a solution with a concentration of 20 mg / mL for the berberine group.

[0159] The preparation method of berberine + black ginger extract gavage solution is as follows: take 1g of berberine + black ginger extract, add appropriate amount of 10% PEG400 saline solution and mix evenly, then add to the total volume of 40mL to prepare a solution with a concentration of 25mg / mL, which is used in the berberine + black ginger extract group.

[0160] Administration method: The rats in the berberine group were given berberine gavage solution by gavage, and the rats in the berberine + black ginger extract group were given berberine + black ginger extract gavage solution by gavage. The administration volume was 10 mL / kg body weight, the administration frequency was 1 time, and the rats were fasted but not watered for about 12 h before administration.

[0161] Blood collection method: Blood was collected from the orbital venous plexus. After administration, the berberine + black ginger extract group repeated the experiment as follows: 0.3 mL of blood was collected from the orbital plexus at 0 min before administration and 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 36 h, 48 h, 60 h, and 72 h after administration, and heparin sodium was used for anticoagulation; centrifuged at 3500 r / min for 10 min, plasma was separated, and frozen at -80 °C.

[0162] (2) Sample content determination

[0163] ①Sample pretreatment method

[0164] Take 50 μl of plasma, add 10 μl of 200 ng / ml bamipine hydrochloride internal standard, add 200 μl of extraction solution 1 (methanol: acetonitrile = 2:1), vortex for 10 min, centrifuge at 4000 rpm for 10 min, take 150 μl of supernatant, blow nitrogen to dryness, and then reconstitute with 200 μl of reconstitution solution 2 (acetonitrile: 0.5% formic acid water = 1:4) for LC-MS detection.

[0165] ②Chromatographic conditions

[0166] Chromatographic column: ACQUITY UPLC BEH C18 (50 mm × 2.1 mm, 1.7 um), Waters Technology Co., Ltd.

[0167] Column temperature: 40°C;

[0168] Mobile phase: A—water (containing 0.1% formic acid), B—methanol (containing 0.1% formic acid);

[0169] Injection volume: 5 μL.

[0170] Table 2

[0171] Time (min) A(％) B(％) Flow rate (mL / min) 0 85 15 0.5 3 70 30 0.5 3.1 20 80 0.5 4 20 80 0.5 4.1 85 15 0.5 5 85 15 0.5

[0172] (3) Mass spectrometry conditions

[0173] The mass spectrometer parameter settings are shown in the table below. High-purity (>99%) nitrogen was used as the ion source gas.

[0174] Table 3

[0175]

[0176] 4. Data processing

[0177] Plotting of blood drug concentration-time curve: The experimental data were processed by GraphPad Prism 6 biostatistics software, and the quantitative data were expressed as mean ± standard error (Mean ± SEM).

[0178] Calculation of pharmacokinetic parameters: The pharmacokinetic parameters of the test samples were calculated using DAS software. The data analysis of the berberine hydrochloride group was based on the results of the first experiment, and the data analysis of the berberine + black ginger extract group was based on the results of repeated experiments.

[0179] Statistical analysis: SPSS 22.0 software was used for significance analysis, and the independent sample T test was used for comparison between the two groups.

[0180] 5. Results and analysis

[0181] (1) Standard curve

[0182] The standard curve was drawn with the concentration of each target compound in the standard as the horizontal axis and the peak area as the vertical axis ( Figure 1 ), the linear regression equation and correlation coefficient of the standard curve are shown in the table below. All target compounds showed good linear relationships within the concentration range investigated, and the correlation coefficient of the linear regression equation (R 2 ) are all greater than 0.99.

[0183] Table 4

[0184] Substance name Linear equations <![CDATA[R 2 ]]> Linear range (ng / ml) Limit of quantitation (ng / ml) Berberine <![CDATA[Y=3.327e 1 X+2222e0]]> 0.9974 0.1～1000 0.1

[0185] (2) Data quality control

[0186] A mixed standard was used as a quality control sample, and its pretreatment method was consistent with that of the calibrator and the sample to be tested. During the detection of every 30 samples, a quality control sample (QC) was interspersed to evaluate the stability of the detection equipment. The concentration of the target compound in the QC sample was obtained by substituting the peak area ratio of each target compound in the QC sample and its corresponding internal standard into the linear regression equation for calculation. If the relative standard deviation (RSD%) of the concentration of all target compounds in the QC sample is less than 20%, and the deviation from the target value is less than 15%, the data quality control is considered to be up to standard. The specific data quality control results are shown in Table 5.

[0187] Table 5

[0188]

[0189]

[0190] (3) Blood drug concentration-time curve

[0191] Blood drug concentration-time curve Figure 2-4 shown.

[0192] (4) Drug metabolism data results (berberine)

[0193] The drug metabolism data results (berberine) are shown in Table 6-7.

[0194] ①Berberine hydrochloride

[0195] Table 6

[0196] Pharmacokinetic parameters unit Mean SD AUC(0-t) ng / mL*min 2614.21 631.336 AUC(0-0) ng / mL*min 5114.298 2795.658 t1 / 2z min 1156.241 1006.976 Tmax min 23.182 12.303 Cmax ng / mL 8.738 4.37

[0197] ②Berberine hydrochloride + black ginger extract

[0198] Table 7

[0199] Pharmacokinetic parameters unit Mean SD AUC(0-t) ng / mL*min 10882.151 4063.551 AUC(0-0) ng / mL*min 11533.94 3919.372 t1 / 2z min 1296.178 764.584 Tmax min 49.091 15.136 Cmax ng / mL 35.809 17.82

[0200] As can be seen from the table, black ginger extract can significantly improve the pharmacokinetic parameters of berberine hydrochloride when used in combination with berberine hydrochloride. Specifically:

[0201] Increasing the values ​​of AUC(0-t) and AUC(0-∞) means increasing the total exposure of the drug in the body. Compared with the use of berberine hydrochloride alone, the combined use of black ginger extract increased AUC(0-t) from 2614.21 ng / mLmin to 10882.151 ng / mLmin, and AUC(0-∞) from 5114.298 ng / mLmin to 11533.94 ng / mLmin.

[0202] Prolonging the t1 / 2z value means prolonging the half-life of the drug. After combined use of black ginger extract, the half-life of berberine hydrochloride was extended from 1156.241min to 1296.178min.

[0203] Increase the Tmax value, that is, prolong the time it takes for the drug to reach maximum concentration. After combined use of black ginger extract, Tmax increased from 23.182min to 49.091min.

[0204] Significantly improve the Cmax value, that is, increase the maximum concentration of the drug in the body. After the combined use of black ginger extract, Cmax increased from 8.738ng / mL to 35.809ng / mL.

[0205] (5) Pharmacokinetic related charts

[0206] Pharmacokinetic related charts such as Figure 5 shown.

[0207] (6) Calculation of bioavailability

[0208] ① Taking the berberine hydrochloride sample as the benchmark, the bioavailability of the black ginger extract group was calculated as follows:

[0209] The black ginger extract group was administered with a dose of 200 mg / kg of berberine, the main ingredient. The bioavailability calculation results of berberine + black ginger extract were:

[0210] F'(berberine+black ginger extract)=(AUCpo berberine+black ginger extract) / (AUCpo berberine hydrochloride)×100%=415.27±185.08%.

[0211] (7) Research conclusions

[0212] This study involved two samples, namely berberine hydrochloride and berberine + black ginger extract.

[0213] According to the research requirements, the two samples were treated with oral gavage respectively, and the berberine concentration in rat plasma was determined by HPLC. The non-compartmental statistical moment analysis model in the pharmacokinetic software DAS was used to calculate the pharmacokinetic parameters and perform pharmacokinetic data analysis to evaluate its relative bioavailability.

[0214] Taking berberine hydrochloride sample as the standard reference preparation, the relative utilization of berberine in berberine + black ginger extract was 415.27±185.08%, showing a significant improvement (P<0.01). After oral administration of berberine + black ginger extract sample and oral berberine hydrochloride, the AUC, Cmax, and Tmax of berberine were significantly different (P<0.01). 1 / 2 There was a certain improvement, but it did not show significance (P>0.05), indicating that the simultaneous oral administration of berberine and black ginger extract can significantly improve the bioavailability of berberine. Compared with the direct oral berberine group, the bioavailability increased by 4.15 times. At the same time, the efficacy of berberine was significantly enhanced, and the in vivo action time was also prolonged. To a certain extent, it solved the problems of poor absorption of berberine in the body, low blood concentration, rapid metabolism, and short half-life. In general, black ginger extract has a significant effect on improving the absorption of berberine.

[0215] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, ordinary technicians in the relevant field should understand that the specific implementation methods of the present invention can still be modified or replaced by equivalents. Any modification or equivalent replacement that does not depart from the spirit and scope of the present invention should be covered within the scope of protection of the claims of the present invention.

Claims

1. A black ginger extract and its application in improving the bioavailability of berberine hydrochloride samples.

2. The use according to claim 1, characterized in that: The preparation method of the black ginger extract comprises: The black ginger is subjected to alcohol extraction and then filtered to obtain a primary extract and an extraction residue, and the extraction residue is subjected to alcohol extraction and filtration again to obtain a secondary extract; the primary extract and the secondary extract are mixed and concentrated to 1 / 3-1 / 5 of the original volume to obtain a concentrated solution; the concentrated solution is adsorbed by a resin to obtain a supernatant, and the supernatant is analyzed by a column using ethanol to obtain a black ginger extract; the black ginger extract is dried to obtain the black ginger extract.

3. The use according to claim 2, characterized in that: The extraction was performed using 50%-60% ethanol solution.

4. The use according to claim 1, characterized in that: The preparation method of berberine hydrochloride comprises: After the piperidine is dissolved in the solvent I, it enters into a condensation process to obtain a condensation product; The condensed product is dissolved in solvent II, a catalyst is added to carry out a hydrogenation reaction, and after the reaction is completed, the catalyst is removed by filtering to obtain a hydrogenated product; The hydrogenation product is mixed with hydrochloric acid to generate a salt compound, and then a condensation reagent is added to carry out a condensation reaction to obtain a condensation product; The condensation product is added to a solvent containing copper salt to carry out a cyclization reaction. During the reaction, a chelating agent is added to remove copper ions. After the cyclization is completed, a crude product is obtained; Dissolve the crude product in hot water, maintain the temperature at 50±5°C and stir; remove insoluble impurities through a filter press to obtain a clear solution; The clear solution is dried to obtain the berberine hydrochloride.

5. The use according to claim 4, characterized in that: The solvent I is anhydrous ethanol; the solvent II is methanol; and the condensation reagent is phosphorus trichloride.

6. The use according to claim 4, characterized in that: The condensation process comprises: The solution of piperonyl ethylamine dissolved in solvent I was cooled to 5° C. under stirring, and a condensation reagent was added dropwise to carry out a condensation reaction, and the reaction time was maintained for at least 1 hour. After the condensation was completed, nitrogen protection was used, the temperature was raised to room temperature, and the condensation product was collected to obtain the condensation product.

7. The use according to claim 6, characterized in that: The condensation reagent is formamide.

8. A product for improving the bioavailability of berberine hydrochloride sample, characterized in that: The product includes black ginger extract.

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