Baicalin ternary co-amorphous drug and preparation method and application thereof

Abstract

The application discloses a baicalin ternary co-amorphous drug and a preparation method and application thereof, and the amorphous substance can effectively improve the solubility and dissolution of puerarin and baicalin. The application mixes metformin hydrochloride, puerarin and baicalin according to a molar ratio of 1:1:1, and obtains the baicalin ternary co-amorphous drug through a ball milling method or a solvent-assisted milling method, the preparation method is green and efficient, the co-amorphous substance has high light stability and thermal stability, the solubility is increased by 1.7 times compared with that of a puerarin raw drug crystal, the solubility is increased by 46.8 times compared with that of a baicalin raw drug crystal, and the solubility is reduced by 14.8 times compared with that of a metformin hydrochloride raw drug crystal. The baicalin ternary co-amorphous drug has certain application prospects in liver protection, blood sugar reduction, treatment of damp-heat type colitis and cervical cancer.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and particularly relates to a baicalin ternary co-amorphous drug and a preparation method and application thereof. BACKGROUND

[0002] Baicalin is a main flavonoid compound in Scutellaria baicalensis Georgi. As a traditional Chinese medicine ingredient, baicalin has very wide pharmacological activities and plays an important role in the prevention and treatment of various diseases, such as anti-inflammatory, anti-tumor, anti-virus, liver protection, and blood sugar reduction. In addition, baicalin also has sunscreen and whitening effects and is widely used in the manufacture of cosmetics.

[0003] Puerarin is a main active ingredient in Pueraria lobata (Willd.) Ohwi and has the effects of reducing blood sugar, clearing heat, resisting cancer, reducing inflammation, and protecting the liver. Studies have shown that puerarin has the effects of enhancing insulin sensitivity and reducing insulin resistance in the prevention and treatment of diabetes (Guo Yongmei, Effects of Different Processed Products of Pueraria lobata on Blood Glucose, China Modern Medicine and Health, 2023, 25(06): 11-14.). At the same time, pueraria and scutellaria form a medicinal pair, which is the main component of the famous Chinese medicine Pueraria lobata Decoction and is commonly used in the treatment of diseases such as diabetes, hyperlipidemia, rectal inflammation of damp-heat type, and cancer.

[0004] Metformin hydrochloride has excellent solubility in water and is a type II diabetes drug widely used in clinical practice, which also has various beneficial biological functions, such as weight loss, anticancer, or treatment of mental diseases.

[0005] A co-amorphous drug is a binary one-way amorphous solid dispersion system formed by mixing a drug active ingredient and other drugs or small molecule excipients. Compared with a crystalline drug, an amorphous drug has greater free energy, higher dissolution rate, and significantly improved solubility. In a co-amorphous system, the two compounds have strong intermolecular interactions due to hydrogen bonding, van der Waals force, etc., so that the glass transition temperature (T g ) of the co-amorphous system is obviously improved, the hygroscopicity is effectively improved, the stability is improved, and the solubility and dissolution of the drug are improved. In addition, the co-amorphous system also has the advantages of realizing the pharmacological synergy between the two drugs or complementary advantages of combined use of drugs.

[0006] Patent CN105497051A discloses a kind of hypoglycemic drug formula and its experimental method, this patent selects 7 active compounds in Gegen Qinlian decoction, flavones derived from kudzu root: puerarin, daidzin;Flavones derived from Huangqin: baicalin, baicalein;Alkaloids derived from Huanglian: berberine;Glycyrrhizic acid and glycyrrhizin derived from licorice, the effect of reducing blood sugar and improving insulin resistance of high blood sugar model rats of 7 compounds with different proportions is compared with the original prescription, and the best hypoglycemic compound composition is obtained: namely puerarin: daidzein: baicalin: baicalein: berberine: glycyrrhizic acid: glycyrrhizin mass ratio is 100:25:100:10:10:5:50.7 Compounds are usually used in the form of crystal. The amorphous state of the compound is not studied. SUMMARY

[0007] In view of the unclear therapeutic effect of the amorphous state of the drug, the present application provides a baicalin ternary co-amorphous drug and a preparation method and application thereof. The preparation method is green and efficient, the co-amorphous drug has high light stability and thermal stability, has better solubility and dissolution rate compared with the puerarin raw material crystal and the baicalin raw material crystal, and effectively delays the dissolution rate of metformin hydrochloride, achieving a sustained-release effect.

[0008] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:

[0009] A baicalin ternary co-amorphous drug, which is mainly composed of metformin hydrochloride, puerarin and baicalin.

[0010] Further, the X-ray diffraction spectrum of the baicalin ternary co-amorphous drug does not have characteristic diffraction peaks of single components, but has characteristic diffuse peaks of amorphous substances.

[0011] Further, the molar ratio of metformin hydrochloride, puerarin and baicalin in the baicalin ternary co-amorphous drug is 1:1:1.

[0012] Further, the preparation method of the baicalin ternary co-amorphous drug comprises the following steps: uniformly mixing metformin hydrochloride, puerarin and baicalin, and then subjecting to ball milling treatment to obtain the baicalin ternary co-amorphous drug.

[0013] Further, the ball milling treatment is carried out at 1000-3000 r / min for 20-40 min.

[0014] Further, the method further comprises adding a solvent before ball milling treatment and drying treatment after ball milling treatment.

[0015] Further, the solvent is at least one of methanol, ethanol and acetonitrile.

[0016] Further, the added amount of the solvent is 15-30 muL of solvent per 1 mmol of metformin hydrochloride.

[0017] The application of the above-mentioned scutellarein ternary co-amorphous drug in the preparation of blood sugar-lowering, liver-protecting, anticancer, and treatment of damp-heat type colitis drugs.

[0018] Further, the cancer is cervical cancer.

[0019] The infrared absorption spectrum measured by the Id7 ATR accessory has absorption peaks at 3322.6, 3193.5, 1687.9, 1653.6, 1610.8, 1571.7, 1448.3, 1354.7, 1294.5, 1248.2, 1211.6, 1059.7, 888.5, 834.1, 795.5, and 725.1 cm -1 The dehydration temperature is 86.2 DEG C.

[0020] The beneficial effects of the present application are:

[0021] 1. The scutellarein ternary co-amorphous drug is prepared for the first time, and the preparation method is green and efficient, can be prepared without solvent medium, has the characteristics of low energy consumption, high economic benefit, and large-scale preparation.

[0022] 2. The balance solubility experiment shows that the scutellarein ternary co-amorphous drug improves the solubility of puerarin by 1.7 times, improves the solubility of scutellarein by 46.8 times, and reduces the solubility of metformin hydrochloride by 14.8 times; the in-vitro dissolution experiment shows that the scutellarein ternary co-amorphous drug improves the dissolution rate of puerarin and scutellarein, and slows down the dissolution rate of metformin hydrochloride, which is helpful for the synergistic enhancement of the three drugs. BRIEF DESCRIPTION OF DRAWINGS

[0023] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor.

[0024] Figure 1 is the powder X-ray diffraction pattern of the scutellarein ternary co-amorphous drug prepared in Example 1.

[0025] Figure 2is a thermal analysis chart of the baicalin ternary co-amorphous drug prepared in Example 1.

[0026] Figure 3 is an infrared spectrum chart of the baicalin ternary co-amorphous drug prepared in Example 1.

[0027] Figure 4 is a stability test result of the baicalin ternary co-amorphous drug prepared in Example 1.

[0028] Figure 5 is a powder X-ray diffraction chart of the baicalin ternary co-amorphous drug prepared in Example 4.

[0029] Figure 6 is a powder X-ray diffraction chart of the baicalin ternary co-amorphous drug prepared in Example 2.

[0030] Figure 7 is a powder X-ray diffraction chart of the baicalin ternary co-amorphous drug prepared in Example 3.

[0031] Figure 8 is a dissolution curve of the baicalin ternary co-amorphous drug prepared in Example 1 in pure water. DETAILED DESCRIPTION

[0032] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative effort belong to the scope of protection of the present application.

[0033] Example 1

[0034] The present embodiment provides a preparation method of a baicalin ternary co-amorphous drug, and the steps are as follows:

[0035] 0.5 mmol of metformin hydrochloride and 0.5 mmol of baicalin and 0.5 mmol of puerarin were placed in a ball mill tank, and after being fully vortexed and uniformly mixed, ball milling was performed, the ball milling time was 30 min, and the ball milling speed was 1500 r / min. The powder after ball milling was collected and placed in a desiccator to obtain the baicalin ternary co-amorphous drug.

[0036] The baicalin ternary co-amorphous drug, puerarin, baicalin and metformin hydrochloride prepared in Example 1 were subjected to powder X-ray diffraction experiment, and the characterization conditions were as follows: a Japanese Rigaku SmartLab X powder X-ray diffractometer was used for analysis. The incident light beam was Cu-Ka ray, the working voltage was 40 kV, the working current was 15 mA, the 2 theta range was 5-50°, and the step was 0.02° / step.

[0037] Crystal form analysis: the XRD crystal form analysis of the obtained amorphous baicalin and the raw materials puerarin, baicalin and metformin hydrochloride is shown in Figure 1 The puerarin has obvious crystal characteristic diffraction peaks at diffraction angles 2 θ : 6.5, 8.0, 8.8, 11.6, 13.9, 15.9, 18.8, 23.4°, etc., the baicalin has crystal characteristic diffraction peaks at diffraction angles 2 θ : 8.62, 12.44, 14.68, 17.02, 20.64, 23.78, 25.44, 27.98°, etc., the metformin hydrochloride has crystal characteristic diffraction peaks at diffraction angles 2 θ : 12.12, 17.56, 19.54, 26.42, 28.14, 29.32, 31.14°, etc., and the baicalin ternary co-amorphous drug prepared in this example presents diffuse diffraction peaks without obvious crystal characteristic diffraction peaks.

[0038] Glass transition temperature test: the baicalin ternary co-amorphous drug, puerarin, baicalin and metformin hydrochloride prepared in this example were subjected to differential scanning calorimetry experiment, and the characterization conditions were as follows: a German Netzsch DSC 214 Nevio differential scanning calorimeter was used for analysis. The same empty crucible was used as a reference, the test temperature range was 30-300℃, the heating rate was 10℃ / min, nitrogen gas was used as protective gas with a flow rate of 40 mL / min.

[0039] The experimental results are shown in Figure 2 The puerarin has an endothermic melting peak at 212.3℃, the baicalin has an endothermic melting peak at 213.6℃, the metformin hydrochloride has an endothermic melting peak at 232.9℃, and the glass transition temperature of the baicalin ternary co-amorphous drug prepared in this example is 128.2℃.

[0040] Infrared test analysis: the baicalin ternary co-amorphous drug, puerarin, baicalin and metformin hydrochloride prepared in Example 1 were subjected to differential scanning calorimetry experiment, and the characterization conditions were as follows: a United States Thermo Nicolet iS 50 attenuated total reflection Fourier infrared spectrometer was used for analysis. The Id7ATR accessory was used for experiment, the scanning number was 32, the resolution was 4cm -1, wavelength range 550-4000 cm -1 .

[0041] The experimental results are shown in Table 1. Figure 3 The O-H characteristic absorption peak of puerarin is at 3367.1 cm -1 , the C=O characteristic absorption peak is at 1620.4 cm -1 , the O-H characteristic absorption peak of baicalin is at 3318.9 cm -1 , the C=O characteristic absorption peak is at 1724.0 cm -1 , the N-H characteristic peak of metformin hydrochloride is at 3366.6 cm -1 , 3287.6 cm -1 , 3142.4 cm -1 , and the C=N stretching vibration peak is at 1621.4 cm -1 . The baicalin ternary co-amorphous drug prepared in Example 1 has absorption peaks at 3322.6, 3193.5, 1687.9, 1653.6, 1610.8, 1571.7, 1448.3, 1354.7, 1294.5, 1248.2, 1211.6, 1059.7, 888.5, 834.1, 795.5 and 725.1 cm -1 . Among them, the O-H peak moves and broadens, indicating that puerarin and baicalin are amorphized during the ball milling process; the blue shift of the C=O and N-H absorption peaks indicates that intermolecular hydrogen bonds may be formed.

[0042] Photothermal stability test analysis: Figure 4 The stability test results of the baicalin ternary co-amorphous drug prepared in Example 1 are shown in Table 2. The light stability test condition is 4500 Lx light intensity (temperature 25 ℃) for 45 days, and the heat stability test condition is 60 ℃ (avoiding light) for 45 days. After the stability test, the sample is re-characterized by powder X-ray diffraction. As shown in Table 2, after the light stability and heat stability tests, the diffraction peaks of the baicalin ternary co-amorphous drug still show the characteristics of diffuse peaks, indicating that the light stability and heat stability of the co-amorphous drug are good. Figure 5

[0043] Example 2

[0044] The present embodiment provides a preparation method of a baicalin ternary co-amorphous drug, the steps are as follows:

[0045] ​Put 0.5 mmol metformin hydrochloride and 0.5 mmol baicalin and 0.5 mmol puerarin into a ball mill tank, fully vortex mix, add 10 μL of methanol, and then ball mill, ball mill time 30 min, ball mill speed 1500 r / min. Collect the ball-milled powder and place it in a desiccator to obtain the baicalin ternary co-amorphous drug.

[0046] X-ray powder diffraction results as shown in Figure 6 The baicalin ternary co-amorphous drug prepared in this example shows diffuse diffraction peaks without crystal characteristic peaks, which is basically the same as that of Example 1.

[0047] Example 3

[0048] This example provides a preparation method of a baicalin ternary co-amorphous drug, the steps of which are as follows:

[0049] Put 0.5 mmol metformin hydrochloride and 0.5 mmol baicalin and 0.5 mmol puerarin into a ball mill tank, fully vortex mix, add 10 μL of methanol, and then ball mill, ball mill time 30 min, ball mill speed 1500 r / min. Collect the ball-milled powder and place it in a desiccator to obtain the baicalin ternary co-amorphous drug.

[0050] X-ray powder diffraction results as shown in Figure 7 The baicalin ternary co-amorphous drug prepared in this example shows diffuse diffraction peaks without crystal characteristic peaks, which is basically the same as that of Example 1.

[0051] Example 4

[0052] This example provides a preparation method of a baicalin ternary co-amorphous drug, the steps of which are as follows:

[0053] Put 0.5 mmol metformin hydrochloride and 0.5 mmol baicalin and 0.5 mmol puerarin into a ball mill tank, fully vortex mix, add 10 μL of methanol, and then ball mill, ball mill time 30 min, ball mill speed 1500 r / min. Collect the ball-milled powder and place it in a desiccator to obtain the baicalin ternary co-amorphous drug.

[0054] X-ray powder diffraction results as shown in Figure 8 The baicalin ternary co-amorphous drug prepared in this example shows diffuse diffraction peaks without crystal characteristic peaks, which is basically the same as that of Example 1.

[0055] Example 5

[0056] This example provides a preparation method of a baicalin ternary co-amorphous drug, the steps of which are as follows:

[0057] 0.5 mmol metformin hydrochloride and 0.5 mmol baicalin and 0.5 mmol puerarin were placed in a ball mill jar, 5 μL of methanol was added, and then the mixture was fully vortexed and ball milled for 20 min at a speed of 3000 r / min. The powder after ball milling was collected and placed in a desiccator to obtain the baicalin ternary co-amorphous drug. The baicalin ternary co-amorphous drug prepared in this example exhibited diffuse diffraction peaks without crystal characteristic peaks, which was basically the same as that of Example 1.

[0058] Example 6

[0059] This example provides a method for preparing a baicalin ternary co-amorphous drug, which comprises the following steps:

[0060] 0.5 mmol metformin hydrochloride and 0.5 mmol baicalin and 0.5 mmol puerarin were placed in a ball mill jar, 15 μL of ethanol was added, and then the mixture was fully vortexed and ball milled for 40 min at a speed of 1000 r / min. The powder after ball milling was collected and placed in a desiccator to obtain the baicalin ternary co-amorphous drug. The baicalin ternary co-amorphous drug prepared in this example exhibited diffuse diffraction peaks without crystal characteristic peaks, which was basically the same as that of Example 1.

[0061] Test Example 1 Amorphous Drug Solubility and Dissolution Test

[0062] Solubility determination:

[0063] An excess amount of metformin hydrochloride, puerarin, baicalin and the baicalin ternary co-amorphous drug prepared in Example 1 were respectively placed in 5 mL centrifuge tubes, 2 mL of pure water was added to obtain a supersaturated solution. The tubes were placed in a 37 °C constant temperature shaker for continuous oscillation for 48 h, the upper clear liquid was taken and filtered with a 0.45 μm membrane, and then immediately diluted to a certain concentration with pure water. The sample concentration was determined by HPLC method to obtain the saturation equilibrium solubility of the sample to be tested.

[0064] The high performance liquid chromatography conditions were as follows: analysis was performed by using a Japan Shimadzu LC-20AD high performance liquid chromatograph. The chromatographic column was a Thermo BDS HYPERSIL C18 chromatographic column (250 mm x 4.6 mm, 5.0 μm); the mobile phase was methanol-0.2% phosphoric acid aqueous solution (30:70); the volume flow rate was 1.0 mL / min; the detection wavelength was 280 nm; the column temperature was 30 °C; the injection amount was 10 μL, and the results are shown in Table 1.

[0065] Table 1 Solubility of each component of the sample in pure water (mg / mL)

[0066]

[0067] It can be seen that the ternary co-amorphous drug of baicalin effectively improves the solubility of puerarin and baicalin, and reduces the solubility of metformin hydrochloride.

[0068] Dissolution test:

[0069] The metformin hydrochloride, puerarin, baicalin and the ternary co-amorphous drug of baicalin prepared in Example 1 were respectively treated by passing through an 80-mesh sieve to avoid the dissolution difference caused by the particle size difference of the powder. 143.22 mg of puerarin, 153.57 mg of baicalin and 353.70 mg of the ternary co-amorphous drug of baicalin were accurately weighed, and then filled into capsules and respectively put into 250 ml of pure water. The average dissolution time of the capsule shell used was about 3 min. Three groups of parallel samples were set, the medium temperature was 37℃, and the rotation speed was 100 r / min. The samples were taken out at 5, 10, 15, 20, 30, 45 min and 1, 2, 4, 8, 12, 24 h, 2 mL of the solution was taken out, and the same temperature and volume of the dissolution medium was supplemented at the same time. The solution was filtered through a 0.45 μm membrane, and the filtrate was appropriately diluted and determined by HPLC method. The results are shown in Table 1. Figure 8 .

[0070] The formation of the ternary co-amorphous drug of baicalin can significantly improve the dissolution of baicalin, keep the dissolution of puerarin basically unchanged, and reduce the dissolution of metformin hydrochloride.

[0071] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A ternary co-amorphous drug of baicalin, characterized in that: The baicalin ternary co-amorphous drug is mainly combined by metformin hydrochloride, puerarin and baicalin; The X-ray diffraction spectrum of the baicalin ternary co-amorphous drug has no characteristic diffraction peak of single component, but has characteristic diffused peak of amorphous substance. The molar ratio of metformin hydrochloride, puerarin and baicalin in the baicalin ternary co-amorphous drug is 1:1:

1.

2. The method of claim 1, wherein the preparation of the ternary co-amorphous drug of baicalin is characterized in that, The method comprises the following steps: The metformin hydrochloride, puerarin and baicalin are uniformly mixed, and the baicalin ternary co-amorphous drug is obtained through ball milling treatment. The ball milling treatment is carried out at 1000-3000 r / min for 20-40 min.

3. The method of claim 2, wherein the preparation of the ternary co-amorphous drug of baicalin is characterized by: The method further comprises adding a solvent before the ball milling treatment and drying treatment after the ball milling treatment.

4. The method of claim 3, wherein the preparation of the ternary co-amorphous drug of baicalin is characterized by: The solvent is at least one of methanol, ethanol and acetonitrile.

5. The method of claim 4, wherein the preparation of the ternary co-amorphous drug of baicalin is characterized by: The adding amount of the solvent is 15-30 μL of solvent per 1 mmol of metformin hydrochloride.

6. The baicalin ternary co-amorphous drug of claim 1 is used for preparing a drug for reducing blood sugar, protecting liver, resisting cervical cancer and treating damp-heat type colitis.

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