WO2018133110A1

WO2018133110A1 - Ziyuglycoside ii solid dispersion and preparation method therefor

Info

Publication number: WO2018133110A1
Application number: PCT/CN2017/072230
Authority: WO
Inventors: 杨世林
Original assignee: 四川英路维特医药科技有限公司
Priority date: 2017-01-23
Filing date: 2017-01-23
Publication date: 2018-07-26

Abstract

A ziyuglycoside II solid dispersion, a preparation method therefor and a use thereof. The solid dispersion is prepared by the following raw and auxiliary materials in weight proportion: 1 part of ziyuglycoside II and 4-30 parts of polyvinylpyrrolidone, and can be used for preparing medicines for increasing the count of blood cells and hemoglobin and for preparing medicines for treating and/or preventing myelosuppression. The solid dispersion can improve the solubility of the ziyuglycoside II and achieves a dissolution rate of 85%.

Description

Earthworm saponin II solid dispersion and preparation method thereof

Technical field

The invention relates to a solid dispersion of saponin II and a preparation method thereof, and belongs to the field of medicine.

Background technique

Myelosuppression is a clinically common hematopoietic disease that can occur in radiation therapy and/or chemotherapy of various systemic neoplastic diseases, radiation damage caused by ionizing radiation, viral hepatitis, parvovirus infection or drugs (chloramphenicol). , benzene, sulfonamides, anti-epileptic drugs, sedatives, anti-thyroid drugs, anti-diabetes drugs, anti-malaria, sleeping pills and other factors. Myelosuppression can cause damage to the bone marrow microenvironment, hematopoietic stem cells, hematopoietic growth factors, etc., and the granulosa, red, and megakaryocyte systems are inhibited. Lack of granulocytes can cause serious infections; a marked reduction in red blood cells can cause severe anemia; a marked drop in platelets causes severe bleeding and even death. At present, there is still no effective treatment for bone marrow suppression in the clinic, and it is urgent to develop a therapeutic drug with better efficacy.

Diltiazem II, chemical name: 3-O-α-L-arabino-19-carboxylic acid (ziyu-glycoside II), is a mantle from the genus Rosaceae or A pharmacologically active compound is extracted from the roots of the longleaf mantle. CN101119740A discloses the use of saponin II in the preparation of a medicament for increasing red blood cells and hemoglobin. However, in actual use, it has been found that saponin II is not effective, and it is often difficult to obtain a better blood cell level and a bone marrow suppression treatment when used alone. Therefore, there is an urgent need to improve the efficacy of the saponin II and promote the clinical application of the drug.

Summary of the invention

It is an object of the present invention to provide a solid dispersion of saponin II and a process for the preparation thereof.

The present invention provides a solid dispersion of saponin II which is prepared from the following raw materials by weight ratio: 1 part of saponin II and 4-30 parts of povidone.

Further, it is prepared from the following raw materials by weight ratio: 1 part of saponin II and 4-10 parts of povidone.

Further, it is prepared from the following raw materials by weight ratio: 1 part of saponin II and 8 parts of povidone.

Further, the povidone is PVP K30.

The invention provides a preparation method of the solid dispersion, comprising the following steps:

a. Dissolving saponin II and povidone in an organic solvent respectively to obtain a solution of saponin II and povidone, and use;

b, adding the saponin II solution to the povidone solution, and mixing uniformly;

c. The organic solvent in the mixture obtained in the step b is removed and pulverized.

Further, the organic solvent described in step a is anhydrous ethanol.

Further, the mass to volume ratio of the saponin II to the solvent in the saponin II solution described in step a is 1:100; the mass to volume ratio of povidone to solvent in the povidone solution is (1: 4) ~ (5:1).

Further, step c was carried out by placing the mixture in a 60 ° C water bath and evaporating the solvent.

The present invention provides the use of the solid dispersion for the preparation of a medicament for increasing the number of blood cells and hemoglobin.

The present invention provides the use of the solid dispersion for the preparation of a medicament for the treatment and/or prevention of myelosuppression.

The present invention provides a method of increasing the number of blood cells and hemoglobin using the solid dispersion.

The present invention provides a method of treating and/or preventing myelosuppression using the solid dispersion. During the research, the inventors found that the reason that the effect of the saponin II on raising the blood cell level is poor is that the solubility is low and the gastrointestinal absorption rate is small, resulting in low bioavailability of the drug and limiting the exertion of its efficacy. The invention uses the povidone as a carrier material to prepare the saponin II as a solid dispersion, which can significantly improve the solubility of the drug, and the dissolution rate of the solid dispersion is as high as 85%. In the pharmacodynamic experiment, compared with the model group, the solid dispersion of the saponin II of the present invention can significantly increase the number of WBC, RBC, PLT, NEUT and HGB in the peripheral blood, and the drug effect is obviously superior to that of the saponin II drug, indicating that the present invention The invention can improve the bioavailability of the main drug and increase the blood cell number and prevent bone marrow suppression by preparing the saponin II into a solid dispersion.

It is apparent that various other modifications, substitutions and changes can be made in the form of the above-described embodiments of the present invention.

The above content of the present invention will be further described in detail below by way of specific embodiments in the form of embodiments. However, the scope of the above-mentioned subject matter of the present invention should not be construed as being limited to the following examples. Any technique implemented based on the above description of the present invention is within the scope of the present invention.

detailed description

The raw materials and equipment used in the specific embodiments of the present invention are known products and are obtained by purchasing commercially available products.

PEG 8000, PEG 6000, PEG 4000, F68 were purchased from Chengdu Kelon Chemical Reagent Factory; PVP K30 was purchased from Sinopharm Chemical Reagent Co., Ltd.

Example 1 Preparation of the solid dispersion of saponin II of the present invention

1.0 g of saponin II was taken and added to 100 ml of absolute ethanol to dissolve to obtain a solution 1. Another 5.0 g of PVP K30 was added, and 20 ml of absolute ethanol was added to ultrasonicate to dissolve to obtain a solution 2. Put solution 2 The mixture was heated to melt on a thermostatic magnetic stirrer while the solution 1 was slowly added to the solution 2 with a dropper. After the solution 1 was all added to the solution 2, the mixture was stirred on a thermostatic magnetic stirrer for four hours. The mixture was poured into an evaporating dish, placed in a 60 ° C water bath, evaporated to dryness, and the dried product was finely ground to obtain a solid dispersion.

Example 2 Preparation of the solid dispersion of saponin II of the present invention

1.0 g of saponin II was taken and added to 100 ml of absolute ethanol to dissolve to obtain a solution 1. Another 8.0 g of PVP K30 was added, and 32 ml of absolute ethanol was added to ultrasonicate to dissolve to obtain a solution 2. Solution 2 was heated to melt on a thermostatic magnetic stirrer while solution 1 was slowly added to solution 2 with a dropper. After solution 1 was all added to solution 2, the mixture was stirred on a thermostatic magnetic stirrer for four hours. The mixture was poured into an evaporating dish, placed in a 60 ° C water bath, evaporated to dryness, and the dried product was finely ground to obtain a solid dispersion.

Example 3 Preparation of the solid dispersion of saponin II of the present invention

1.0 g of saponin II was taken and added to 100 ml of absolute ethanol to dissolve to obtain a solution 1. Another 20.0 g of PVP K30 was added to the solution by adding 4 ml of absolute ethanol to dissolve to obtain a solution 2. Solution 2 was heated to melt on a thermostatic magnetic stirrer while solution 1 was slowly added to solution 2 with a dropper. After solution 1 was all added to solution 2, the mixture was stirred on a thermostatic magnetic stirrer for four hours. The mixture was poured into an evaporating dish, placed in a 60 ° C water bath, evaporated to dryness, and the dried product was finely ground to obtain a solid dispersion.

The beneficial effects of the present invention are exemplified below by experiments.

Dissolution method: Take one tablet of each batch, according to the dissolution method ("Chinese Pharmacopoeia" Appendix XC second method), with pH 6.8PBS 1 000mL as the dissolution medium, the rotation speed is 100r·min ^-1 , temperature ( 37±0.5)°C, sample 5mL at 0, 5, 15, 20, 25min, filter with 0.45μm microporous membrane, place the filtrate in a 100mL volumetric flask, dilute to the mark with pH 6.8PBS, shake well, UV- Visible spectrophotometry (Chinese Pharmacopoeia Appendix IVA) measured absorbance at 345nm wavelength; after 25min sampling, the whole solution was transferred to a beaker, heated and stirred at 100 ° C for 2 ~ 3min, cooled to 37 ° C, according to the above method After sampling and dilution, the absorbance measured by the eluate after treatment at 100 ° C was the denominator, and the absorbance measured at each time point was taken as a molecule, and the cumulative dissolution rate was calculated.

Experimental Example 1 Quality Evaluation of Preparation of Diosgenin II Solid Dispersion Using Different Carrier Materials

Five experimental groups were set up in this experiment. 1.0 g of saponin II was separately taken and ultrasonically added to 100 ml of anhydrous ethanol to dissolve to obtain a solution 1. Another 8.0g of different polymer materials PVP K30, poloxamer 188 (F68), polyethylene glycol (PEG) 8000, polyethylene glycol (PEG) 6000, polyethylene glycol (PEG) 4000, add 4ml The anhydrous ethanol was sonicated to dissolve to obtain a solution 2. Solution 2 was heated to melt on a thermostatic magnetic stirrer while solution 1 was slowly added to solution 2 with a dropper. After solution 1 was all added to solution 2, the mixture was stirred on a thermostatic magnetic stirrer for four hours. The mixture was poured into an evaporating dish, placed in a 60 ° C water bath, evaporated to dryness, and the dried product was finely ground to obtain a solid dispersion. The solubility and dissolution of the solid dispersion were measured, and the results are shown in Table 1.

Table 1 Determination of Solubility and Dissolution of Diquat Saponin II Solid Dispersion

The solubility of the saponin II drug in the early stage of the experiment was determined to be only 0.032 mg/ml. It can be seen from Table 1 that when the solid dispersion of the saponin II is prepared by using the povidone-based carrier material of the present invention, the solubility of the drug can be significantly improved, and the dissolution degree is highest; if other carrier materials, such as poloxamer, are used, With polyethylene glycol and the like, the dissolution rate of the solid dispersion is greatly reduced, and the solubility of the drug is not as obvious as that of PVP K30.

The above results show that the quality of the solid dispersion of saponin II prepared by using povidone as the carrier material is the best.

Experimental Example 2 Effect of the Amount of Support Material on the Quality of Diquat Saponin II Solid Dispersion

Six experimental groups were set up in this experiment. 1.0 g of saponin II was separately taken and ultrasonically added to 100 ml of anhydrous ethanol to dissolve to obtain a solution 1. Further, PVP K30 (1 part by mass of saponin II and 1 part by mass of PVP K30) was weighed according to the mass ratio shown in Table 2, and ultrasonically dissolved in 4 ml of anhydrous ethanol to dissolve to obtain a solution 2. Solution 2 was heated to melt on a thermostatic magnetic stirrer while solution 1 was slowly added to solution 2 with a dropper. After solution 1 was all added to solution 2, the mixture was stirred on a thermostatic magnetic stirrer for four hours. The mixture was poured into an evaporating dish, placed in a 60 ° C water bath, evaporated to dryness, and the dried product was finely ground to obtain a solid dispersion. The solubility and dissolution of the solid dispersion were measured, and the results are shown in Table 2.

Table 2 Determination of Solubility and Dissolution of Diquat Saponin II Solid Dispersion

Experimental results: When the amount of PVP K30 as carrier material is 4-30 times of saponin, a good solid dispersion can be obtained: the solubility of the drug is increased to 0.594 mg/mL, the dissolution is more than 64%; the amount of PVP K30 is mantle. When the saponin II is 4-10 times, the quality of the preparation is further optimized, the solubility of the drug is increased to 0.4 mg/mL or more, and the dissolution is not less than 70%; wherein the mass ratio of the saponin II: PVP K30 is 1:8. The prepared solid dispersion has the highest solubility and dissolution.

Experimental Example 3 Efficacy Experiment of Diquat Saponin II Solid Dispersion

1. Experimental materials, reagents and instruments

1.1. Test drug: saponin II solid dispersion (prepared according to Example 2), saponin II.

1.2, tool drugs: cyclophosphamide.

1.3. Experimental animal KM-mouse: 18.5-22.5 g.

1.4, experimental equipment: automatic blood cell analyzer; BS-600L electronic balance: specifications: 600g / 0.1g, Shanghai Yousheng Weighing Apparatus Co., Ltd.

2, statistical methods

Statistical analysis was performed using SPSS 17.0 software. Data in mean ± standard deviation (

), one-way analysis of variance was used between groups, LSD test was performed between groups with variance, and Tamhane's T2 test was performed for those with irregular variance.

3. Experimental methods

3.1, experimental animal grouping and model preparation

All animals were fed ad libitum for 1 week and were randomly divided into: blank group; model group; saponin II solid dispersion group, which were formulated into 0.5 mg·kg ^-1 , 5 mg·kg ^-1 , 10 mg·kg ^{-1 .} The suspension was prepared before use; the saponin II group was formulated into a 10 mg·kg ^-1 suspension and prepared before use. On the first day of the experiment, except for the blank group, the other groups of mice were intraperitoneally injected with cyclophosphamide physiological saline solution at a dose of 50 mg·kg ^-1 for 3 consecutive days, and the blank mice were injected with the same volume of normal saline in the tail vein.

3.2, administration

The experimental groups were given the corresponding drugs by dose and gavage from the first day of the experiment. The mice in the blank group and the model group were intragastrically administered with the same volume of normal saline for 7 consecutive days.

3.3, specimen collection

On the 8th day of the experiment, blood was taken from the eye of each experimental group and collected with a 0.5 ml EP tube containing EDTA anticoagulant.

3.4. Test indicators and methods

Peripheral blood test: Peripheral blood leukocytes (WBC), neutrophils (NEUT) red blood cells (RBC), platelets (PLT), and hemoglobin (HGB) were counted in each experimental group by an automatic blood cell counter.

4. Experimental results

Table 3 Results of peripheral blood test results of mice in each experimental group

Note: Compared with the model group, *P<0.05, **P<0.01; compared with the saponin II group, △P<0.05, △△P<0.01.

As can be seen from Table 3, compared with the model group, the number of WBC, RBC and PLT in the peripheral blood of each group of the saponin II solid dispersion was significantly increased (P<0.05), and there was no significant difference in the saponin II group. Compared with the saponin II group, the number of WBC, RBC and PLT in the peripheral blood of the mice in the solid dispersion of saponin II was significantly increased (P<0.05).

Table 4 Results of peripheral blood test results of mice in each experimental group

As can be seen from Table 4, compared with the model group, the NEUT and HGB numbers in the peripheral blood of the saponin II solid dispersion group were significantly increased (P<0.05), and there was no significant difference in the saponin II group; Compared with the saponin II group, the NEUT and HGB numbers in the peripheral blood of the saponin II solid dispersion group were significantly increased (P<0.05).

The above experimental results show that the preparation of the saponin II as a solid dispersion according to the formulation of the present invention can significantly improve the blood cell level of the drug, and can effectively prevent bone marrow suppression.

Claims

A solid dispersion of saponin II characterized in that it is prepared from the following raw materials by weight ratio: 1 part of saponin II and 4-30 parts of povidone.
The solid dispersion according to claim 1, which is prepared from a raw material of the following weight ratio: 1 part of saponin II and 4 to 10 parts of povidone.
The solid dispersion according to claim 2, which is prepared from the following raw materials by weight ratio: 1 part of saponin II and 8 parts of povidone.
The solid dispersion according to any one of claims 1 to 3, wherein the povidone is PVP K30.
A method for preparing a solid dispersion according to any one of claims 1 to 4, which comprises the steps of:

a. Dissolving saponin II and povidone in an organic solvent respectively to obtain a solution of saponin II and povidone, and use;

b, adding the saponin II solution to the povidone solution, and mixing uniformly;

c. The organic solvent in the mixture obtained in the step b is removed and pulverized.
The preparation method according to claim 5, wherein the organic solvent in the step a is anhydrous ethanol.
The preparation method according to claim 5 or 6, wherein the mass ratio of the saponin II to the solvent in the solution of the saponin II in the step a is 1:100; in the povidone solution The mass to volume ratio of povidone to solvent is (1:4) to (5:1).
The process according to claim 5, wherein the step c is carried out by placing the mixture in a 60 ° C water bath and evaporating the solvent.
The solid dispersion according to any one of claims 1 to 4 for preparing elevated blood cells and blood red eggs Use in white quantities of drugs.
Use of the solid dispersion according to any one of claims 1 to 4 for the preparation of a medicament for the treatment and/or prevention of myelosuppression.
A method for raising the number of blood cells and hemoglobin, which comprises using the solid dispersion according to any one of claims 1 to 4.
A method for treating and/or preventing myelosuppression, which comprises using the solid dispersion according to any one of claims 1 to 4.