Wogonin and oroxin b co-amorphous material and application thereof

Abstract

The application discloses a wogonoside and matrine co-amorphous substance and application thereof, and belongs to the technical field of pharmacy. The wogonoside and matrine co-amorphous substance is prepared by using wogonoside and matrine in a molar ratio of 1:2 to 2:1. The application has the following advantages: the solubility of the wogonoside and matrine co-amorphous substance is about 40 times higher than that of wogonoside. Wogonoside has a rigid planar structure, and solvent molecules are difficult to penetrate the structure due to the close arrangement of molecules, resulting in poor solubility. After wogonoside and matrine form a co-amorphous substance, the close structure of the molecules is destroyed, and solvent molecules can penetrate the structure, so that the solubility is improved.

Description

Technical Field

[0001] This invention relates to the amorphous compound of baicalin and strychnine and its applications, belonging to the field of pharmaceutical technology. Background Technology

[0002] Baicalein (5,7-dihydroxy-8-methoxyflavonoid) is one of the main active ingredients isolated from the traditional Chinese medicine Scutellaria baicalensis. It possesses various pharmacological activities, including antitumor, anti-inflammatory, antioxidant, antibacterial, antiviral, anticonvulsant, anti-anxiety, and neuroprotective effects. However, baicalein has poor solubility, with an oral bioavailability of only 1.1%, resulting in blood concentrations that do not meet the requirements for clinical treatment.

[0003] To address the poor solubility of baicalin, researchers have explored various methods to improve it, including structural modification and alteration through chemical methods, and dosage form modification using polymeric materials as drug carriers. While these methods have improved the solubility of baicalin to some extent, they face significant limitations in actual industrial production. For example, structural modification can result in poor reaction selectivity and low yield, and using formulation strategies to increase solubility may lead to reduced drug permeability. Summary of the Invention

[0004] The technical problem to be solved by this invention is to prepare a co-amorphous form of baicalin and strychnine to improve the solubility of baicalin, thereby increasing its bioavailability and providing a research basis for its new drug development.

[0005] To achieve the above objectives, the present invention adopts the following technical solution.

[0006] The amorphous compound of baicalein and physostigmine was prepared by using baicalein and physostigmine in a molar ratio of 1:2 to 2:1.

[0007] Preferably, the amorphous compound of baicalein and physostigmine is prepared by using baicalein and physostigmine in a molar ratio of 1:1.

[0008] The amorphous compound of baicalin and physostigmine is prepared as follows: baicalin and physostigmine in a molar ratio of 1:2 to 2:1 are added to an appropriate amount of deionized water and ground until the solvent evaporates; or the mixture is stirred for 24 to 72 hours, filtered, and the filter cake is dried in a 100°C oven to obtain the final product.

[0009] The preparation method is as follows: add baicalein and physostigmine in a molar ratio of 1:2 to 2:1 to an appropriate amount of deionized water, grind until the solvent evaporates; or stir for 24 to 72 hours, filter, and dry the filter cake to obtain the final product.

[0010] Preferably, the preparation method is any one of the following:

[0011] Method 1: Weigh 28.426 mg of baicalein (0.1 mmol) and 23.236 mg of physostigmine (0.1 mmol) and place them in a clean mortar. Add 200 μL of deionized water and grind until the solvent evaporates.

[0012] Method 2: Weigh 284.26 mg baicalein (1 mmol) and 232.36 mg matrine (1 mmol) into a 100 mL Erlenmeyer flask, add 3 mL of deionized water, and stir continuously on a magnetic stirrer for 48 h at a speed of 400 rpm. After filtration, dry the filter cake in an oven at 100 °C for 60 min.

[0013] Method 3: Weigh 28.426g of baicalein (0.1mol) and 23.236g of physostigmine (0.1mol), place them in a 100mL Erlenmeyer flask, add 10mL of deionized water, and stir continuously on a magnetic stirrer for 48h at a speed of 400rpm. After filtration, dry the filter cake in a 100℃ oven for 60min.

[0014] This invention is based on the following research findings:

[0015] The inventors analyzed the molecular structure of baicalein and found that there is one phenolic hydroxyl group at the 5th and 7th positions, which can act as a proton donor. The carbonyl group at the 4th position has an electron-withdrawing effect and can act as a proton acceptor. Therefore, the phenolic hydroxyl group can form intermolecular hydrogen bonds with groups such as amide, carboxyl, and pyridine, and the carbonyl group can form intermolecular hydrogen bonds with groups such as hydroxyl and carboxyl.

[0016] Based on structural analysis and the rules governing the formation of non-covalent bonds, we searched the FDA database (GRAS, generally regarded as safe), EAFUS (everything added to food in the United States), and the Cambridge Compound Database. We selected various pharmaceutically acceptable substances that could theoretically form hydrogen bonds or other non-covalent bonds with baicalein as ligands, such as matrine, styraxine, sophoridine, sophoridine, cytisine, tetramethylpyrazine, theophylline, and matrine. We used solution suspension, liquid-addition grinding, and recrystallization methods to investigate the reaction results of baicalein and ligands in different solvents at different molar ratios (1:1, 1:2, 2:1).

[0017] Preliminary experiments revealed that only strychnine could form a 1:1 co-amorphous compound with baicalein, and this co-amorphous compound significantly improved the solubility of baicalein. More importantly, strychnine also possesses pharmacological activities such as antitumor and antipulmonary hypertension; the formation of the co-amorphous compound may produce a positive synergistic effect, thereby increasing the efficacy. Therefore, a co-amorphous compound of baicalein and strychnine was prepared.

[0018] In the research and development experiments, the inventors tried different ratios of baicalein: physostigmine (1:2) and baicalein: physostigmine (2:1) through screening experiments. They found that when the molar ratio of the two was 1:1, there was no residue of each of the raw materials. Therefore, this ratio was selected as the optimal ratio for preparation.

[0019] In a second aspect, the present invention provides a method for preparing amorphous baicalin and physostigmine.

[0020] The preparation method of the amorphous compound of baicalin and physostigmine involves adding baicalin and physostigmine in a molar ratio of 1:2 to 2:1 to an appropriate amount of deionized water, grinding until the solvent evaporates; or stirring for 24 to 72 hours, filtering, and drying the filter cake to obtain the product.

[0021] Preferably, the method for preparing the amorphous compound of baicalin and physostigmine involves adding baicalin and physostigmine in a molar ratio of 1:1 to an appropriate amount of deionized water, grinding until the solvent evaporates; or stirring for 48 hours, filtering, and drying the filter cake to obtain the final product.

[0022] More preferably, the preparation method of the amorphous compound of baicalin and physostigmine is any one of the following methods:

[0023] Method 1: Weigh 28.426 mg of baicalein (0.1 mmol) and 23.236 mg of physostigmine (0.1 mmol) and place them in a clean mortar. Add 200 μL of deionized water and grind until the solvent evaporates.

[0024] Method 2: Weigh 284.26 mg baicalein (1 mmol) and 232.36 mg matrine (1 mmol) into a 100 mL Erlenmeyer flask, add 3 mL of deionized water, and stir continuously on a magnetic stirrer for 48 h at a speed of 400 rpm. After filtration, dry the filter cake in an oven at 100 °C for 60 min.

[0025] Method 3: Weigh 28.426g of baicalein (0.1mol) and 23.236g of physostigmine (0.1mol), place them in a 100mL Erlenmeyer flask, add 10mL of deionized water, and stir continuously on a magnetic stirrer for 48h at a speed of 400rpm. After filtration, dry the filter cake in a 100℃ oven for 60min.

[0026] A third aspect of the present invention provides the use of baicalin and physostigmine in amorphous form.

[0027] The use of baicalein and strychnine in the preparation of drugs for treating tumors.

[0028] The use of baicalein and strychnine in the preparation of drugs for treating pulmonary hypertension.

[0029] The use of baicalein and strychnine in the preparation of neuroprotective drugs.

[0030] Baicalein not only possesses various pharmacological activities such as antitumor, anti-anxiety, and neuroprotective effects, but also exhibits significant effects in delaying ventricular hypertrophy and reducing aortic contraction. Furthermore, physostigmine also possesses antitumor and cardiovascular protective pharmacological activities. Therefore, the amorphous compound of baicalein and physostigmine prepared in this invention is intended for the treatment of related diseases such as antitumor activity, anti-pulmonary hypertension, and neuroprotection.

[0031] The advantages of this invention are as follows: Compared to baicalin, the solubility of the co-amorphous compound of baicalin and physostigmine is improved by approximately 40 times. Baicalin has a rigid planar structure with tightly packed molecules, making it difficult for solvent molecules to penetrate its structure, resulting in poor solubility. After baicalin and physostigmine form a co-amorphous compound, the tight molecular structure is disrupted, allowing solvent molecules to penetrate its structure, thereby improving solubility. Attached Figure Description

[0032] The specific embodiments of the present invention will be described in further detail below with reference to the accompanying drawings.

[0033] Figure 1 X-ray diffraction patterns of baicalein (wog), alop, and amorphous powders of baicalein and alop (wog-alop).

[0034] Figure 2 Differential scanning calorimetry (DSC) images of baicalein (wog), alop, and the co-amorphous form of baicalein and alop (wog-alop).

[0035] Figure 3 The states of baicalein (wog, 1 mg / mL) and baicalein and physostigmine co-amorphous form (wog-alop, equivalent to 1 mg / mL of baicalein) after dissolution.

[0036] Figure 4 Solubility of baicalein (wog) and baicalein-alop co-amorphous form (wog-alop) in aqueous solution at pH 7.0 Detailed Implementation

[0037] In the amorphous form of baicalin and physostigmine, the molar ratio of baicalin to physostigmine is 1:1. Their molecular formulas and structural formulas are as follows:

[0038] ①Scutellaria baicalensis

[0039] Molecular formula: C 16 H 12 O5

[0040] Structural formula:

[0041]

[0042] ②Stichope

[0043] Molecular formula: C 15 H 24 N2

[0044] Structural formula:

[0045]

[0046] Baicalein: Purchased from Hubei Wande Chemical Co., Ltd., purity 98%.

[0047] Matrine: Purchased from Shanghai Yuanye Biotechnology Co., Ltd., purity 98%.

[0048] Example 1: Preparation of amorphous compound of baicalin and physostigmine

[0049] Weigh 28.426 mg of baicalein (0.1 mmol) and 23.236 mg of physostigmine (0.1 mmol). Place the baicalein and physostigmine in a clean mortar, add 200 μL of deionized water, and grind until the solvent evaporates.

[0050] Example 2: Preparation of amorphous compound of baicalin and physostigmine

[0051] Weigh 284.26 mg of baicalein (1 mmol) and 232.36 mg of physostigmine (1 mmol). Place the baicalein and physostigmine in a 100 mL Erlenmeyer flask, add 3 mL of deionized water, and stir continuously on a magnetic stirrer for 48 h at a speed of 400 rpm. After filtration, dry the filter cake in an oven at 100 °C for 60 min.

[0052] Example 3: Preparation of amorphous compound of baicalin and physostigmine

[0053] Weigh 28.426g of baicalein (0.1mol) and 23.236g of physostigmine (0.1mol). Place the baicalein and physostigmine in a 100mL Erlenmeyer flask, add 10mL of deionized water, and stir continuously on a magnetic stirrer for 48h at a speed of 400rpm. After filtration, dry the filter cake in an oven at 100℃ for 60min.

[0054] Example 4: Preparation of amorphous compound of baicalin and physostigmine

[0055] Weigh 56.852 mg of baicalein (0.2 mmol) and 23.236 mg of maltol (0.1 mmol). Place the baicalein and maltol in a clean mortar, add 400 μL of deionized water, and grind until the solvent evaporates.

[0056] Example 5: Preparation of amorphous compound of baicalin and physostigmine

[0057] Weigh 28.426 mg of baicalein (0.1 mmol) and 46.472 mg of physostigmine (0.2 mmol). Place the baicalein and physostigmine in a clean mortar, add 400 μL of deionized water, and grind until the solvent evaporates.

[0058] Example 6: Preparation of amorphous compound of baicalin and physostigmine

[0059] Weigh 284.26 mg of baicalein (1 mmol) and 232.36 mg of physostigmine (1 mmol). Place the baicalein and physostigmine in a 100 mL Erlenmeyer flask, add 3 mL of deionized water, and stir continuously on a magnetic stirrer for 24 h at a speed of 400 rpm. After filtration, dry the filter cake in an oven at 100 °C for 60 min.

[0060] Example 7: Preparation of amorphous compound of baicalin and physostigmine

[0061] Weigh 28.426g of baicalein (0.1mol) and 23.236g of physostigmine (0.1mol). Place the baicalein and physostigmine in a 100mL Erlenmeyer flask, add 10mL of deionized water, and stir continuously on a magnetic stirrer for 72h at a speed of 400rpm. After filtration, dry the filter cake in an oven at 100℃ for 60min.

[0062] Example 8: X-ray diffraction

[0063] I. Instruments and Reagents

[0064] Baicalein: Purchased from Hubei Wande Chemical Co., Ltd., purity 98%.

[0065] Matrine: Purchased from Shanghai Yuanye Biotechnology Co., Ltd., purity 98%.

[0066] Amorphous compound of baicalin and physostigmine: prepared according to Example 1

[0067] Test instruments:

[0068] Name: Powder X-ray Diffractometer

[0069] Model: Rigaku D / max-2550

[0070] Manufacturer: Rigaku Corporation, Japan

[0071] II. Experimental Methods

[0072] The powder X-ray diffraction conditions were as follows: CuKa radiation, graphite monochromator, tube voltage 40 kV, tube current 150 mA, 2θ scan range 3–40°, scan speed 8° / min, step size 0.02°, divergence slit DS = 1°, receiving slit RS = 0.15 mm, and scattering slit SS = 1°. Under the above experimental conditions, the powder X-ray diffraction patterns of baicalin, physostigmine, and amorphous products of baicalin and physostigmine were accurately weighed.

[0073] III. Experimental Results

[0074] The results are as follows Figure 1 As shown. By Figure 1 It can be seen that after baicalin and physostigmine form a co-amorphous powder, the characteristic diffraction peaks of baicalin and physostigmine disappear, and a typical co-amorphous powder X-ray diffraction pattern is generated.

[0075] Example 9: Differential Scanning Calorimetry

[0076] I. Instruments and Reagents

[0077] Baicalein: Purchased from Hubei Wande Chemical Co., Ltd., purity 98%.

[0078] Matrine: Purchased from Shanghai Yuanye Biotechnology Co., Ltd., purity 98%.

[0079] Amorphous compound of baicalin and physostigmine: prepared according to Example 1

[0080] Test instruments:

[0081] Name: Differential Scanning Calorimeter

[0082] Model: DSC1

[0083] Manufacturer: Mettler Toledo, Switzerland

[0084] II. Experimental Methods

[0085] 5 mg of baicalein, physostigmine, and amorphous mixture of baicalein and physostigmine were accurately weighed and placed in an aluminum crucible, with an empty aluminum crucible as a reference. The heating rate was 10 K / min and the N2 flow rate was 50 mL / min. The differential scanning calorimetry (DSC) spectra of each were measured.

[0086] III. Experimental Results

[0087] The results are as follows Figure 2 As shown. By Figure 2 It can be seen that the differential scanning calorimetry (DSC) of the amorphous mixture of baicalin and physostigmine does not show the endothermic peaks of baicalin and physostigmine, further indicating that the two have formed a new phase.

[0088] Example 10: Solubility Experiment

[0089] I. Instruments and Reagents

[0090] Baicalein: Purchased from Hubei Wande Chemical Co., Ltd., purity 98%.

[0091] Matrine: Purchased from Shanghai Yuanye Biotechnology Co., Ltd., purity 98%.

[0092] Amorphous compound of baicalin and physostigmine: prepared according to Example 1

[0093] Test instruments:

[0094] Name: Ultrasonic Cleaner

[0095] Model: S30H

[0096] Manufacturer: Elma GmbH, Germany

[0097] II. Experimental Methods

[0098] Accurately weigh 10 mg of baicalein or an equivalent amount of baicalein and physostigmine amorphous compound, place it in a 20 mL vial, add 10 mL of pure water, and sonicate for 1 min to prepare a 1 mg / mL solution of baicalein or baicalein and physostigmine amorphous compound.

[0099] III. Experimental Results

[0100] The results are as follows Figure 3 As shown. By Figure 3 It is evident that, at the same concentration, the solubility of the co-crystallization of baicalein and strychnine in amorphous form is far superior to that of baicalein, indicating that the co-crystallization technique has successfully improved the solubility of baicalein.

[0101] Example 11: In vitro dissolution experiment

[0102] I. Instruments and Reagents

[0103] Baicalein: Purchased from Hubei Wande Chemical Co., Ltd., purity 98%.

[0104] Matrine: Purchased from Shanghai Yuanye Biotechnology Co., Ltd., purity 98%.

[0105] Amorphous compound of baicalin and physostigmine: prepared according to Example 1

[0106] Test instruments:

[0107] Name: Intelligent Dissolution Tester

[0108] Model: RC12AD

[0109] Manufacturer: Tianjin Tianda Tianfa Technology Co., Ltd.

[0110] II. Experimental Methods

[0111] Accurately weigh 60 mg of baicalein or an equivalent amount of baicalein and physostigmine amorphous compound, add to 900 mL of pure water, and conduct an in vitro dissolution experiment using a paddle method at 37 °C and 100 r / min. Samples were taken at 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 30, 45, and 60 min. The samples were filtered through a 0.22 μm microporous membrane and detected by high performance liquid chromatography (mobile phase: acetonitrile:water (55:45), flow rate: 1.0 mL / min; column temperature: 30 °C; injection volume: 10 μL; detection wavelength: 275 nm; chromatographic column: Agilent ZORBAX SB-C18 (4.6 × 250 mm, 5 μm)). The content was calculated using the standard curve method. Plot the solubility curves of baicalein and its amorphous compound with time (min) on the x-axis and the solubility (%) on the y-axis, respectively. (See attached figures.) Figure 4 .

[0112] III. Experimental Results

[0113] The results are as follows Figure 3 and Figure 4 As shown. By Figure 3 It is evident that, at the same concentration, the solubility of the co-crystallization of baicalein and strychnine in amorphous form is far superior to that of baicalein, indicating that the co-crystallization technique has successfully improved the solubility of baicalein. Figure 4 The solubility of baicalein (wog) and the amorphous form of baicalein and physostigmine (wog-alop) in aqueous solution at pH 7.0 is shown. The data indicate that the solubility of the amorphous form of baicalein and physostigmine (wog-alop) in water is significantly higher than that of baicalein.

[0114] The reason for this is that baicalein has a rigid planar structure with tightly packed molecules, making it difficult for solvent molecules to penetrate, resulting in poor solubility. This invention forms a co-amorphous compound of baicalein and phytic acid, which disrupts the tight molecular structure, allowing solvent molecules to penetrate and thus greatly improving solubility.

[0115] The detailed descriptions listed above are merely specific illustrations of feasible embodiments of the present invention and are not intended to limit the scope of protection of the present invention. Those skilled in the art can devise many other modifications and embodiments, which will fall within the scope of this application. More specifically, within the scope of this application, the drawings, and the claims, various variations and improvements can be made to the components and / or layout of the subject matter combination layout. Besides variations and improvements to the components and / or layout, other uses will be apparent to those skilled in the art.

Claims

1. An amorphous compound of baicalin and physostigmine, characterized in that: It was prepared by using baicalein and physostigmine in a molar ratio of 1:1, and the preparation method is as follows: Weigh 0.1 mmol of baicalein and 0.1 mmol of physostigmine and place them in a clean mortar. Add 200 μL of deionized water and grind until the solvent evaporates. The molar ratio of baicalein to physostigmine in the co-amorphous product is 1:

1.

2. The method for preparing the amorphous compound of baicalin and physostigmine according to claim 1, characterized in that, The preparation method is as follows: Weigh 0.1 mmol of baicalein and 0.1 mmol of physostigmine and place them in a clean mortar. Add 200 μL of deionized water and grind until the solvent evaporates.

3. The use of the amorphous compound of baicalein and strychnine as described in claim 1 in the preparation of a drug for treating tumors.

4. The use of the amorphous compound of baicalein and strychnine as described in claim 1 in the preparation of a drug for treating pulmonary hypertension.

5. The use of the amorphous compound of baicalein and strychnine as described in claim 1 in the preparation of a neuroprotective drug.

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