WO2015113369A1

WO2015113369A1 - Stevioside a glycoside crystal, preparation method therefor, and uses thereof

Info

Publication number: WO2015113369A1
Application number: PCT/CN2014/080908
Authority: WO
Inventors: 朱理平; 梅雪峰
Original assignee: 诸城市浩天药业有限公司
Priority date: 2014-01-30
Filing date: 2014-06-27
Publication date: 2015-08-06
Also published as: CN103739639A; CN103739639B

Abstract

Disclosed is a stevioside A glycoside crystal form 9, of which the structure is represented by the formula I. The X-ray powder derivative (XRPD) figure of the crystal form 9 has the following characteristic peaks at the angles of 2θ±0.1 degrees: 4.748, 9.511, 11.767, 13.598, 14.038, 17.224, 19.242, and 23.428.

Description

Stevioside A glycoside crystal, preparation method and use thereof

Technical field

The invention relates to the field of chemical pharmacy, in particular to a novel stevioside A glycoside crystal, a preparation method and use thereof. Background technique

The polymorphism phenomenon refers to a phenomenon in which a solid matter is arranged in two or more different spatial arrangements to form a solid state having different physicochemical properties. In the field of pharmaceutical research, polymorphs include multi-component crystal forms such as organic solvates, hydrates, and the like. Drug polymorphism is widespread in drug development and is an inherent property of organic small molecule compounds. Theoretically, small molecule drugs can have an infinite number of crystal packing methods—polymorphs. Studies have shown that the number of drug polymorphs found is directly proportional to the time and resources of the research they are investigating. Like Lipitor, the world's highest-selling drug to date, there are as many as 35 patents for patent protection. Polymorphism is not only controlled by internal factors such as the spatial structure and functional group properties of molecules, intramolecular and intermolecular interactions, but also by drug synthesis process design, crystallization and purification conditions, formulation excipient selection, and formulation process. Route and granulation methods, as well as storage conditions, packaging materials and other factors. Different crystal forms have different colors, melting points, dissolution, dissolution properties, chemical stability, reactivity, mechanical stability, etc. These physicochemical properties or processability sometimes directly affect the safe and effective performance of the drug. Therefore, crystal research and control has become an important research content in the drug development process.

Crystallization studies include two stages of crystal discovery and crystal form optimization. In the crystal discovery stage, a variety of crystallization methods are used, such as melt crystallization, solution evaporation, rapid cooling and suspension crystallization, by changing the crystallization conditions, solvent , external factors affecting the crystallization of the drug, such as temperature, speed and ratio of suspended solvent. High-throughput sample preparation platform is used to prepare hundreds of crystallization tests simultaneously, using micro sample preparation techniques and analytical testing methods. New crystal forms were prepared and discovered. In the preferred stage of the crystal form, for the new crystal form process amplification and preparation conditions, a variety of solid characterization methods, such as X-ray diffraction, solid state nuclear magnetic resonance, Raman spectroscopy, infrared spectroscopy, etc. In addition, the physicochemical properties of the crystal form should be studied by DSC, TGA, DVS, HPLC, etc., and the hygroscopicity, chemical stability, physical state stability, and processability of different crystal forms were compared. Finally, the most preferred solid form is selected for development.

Stevioside A glycoside is a kauriene diterpene glycoside, a leaf from the asteraceae herb stevia A new natural sweetener from Zhongjing, native to Brazil and Paraguay. According to the international sweetener industry, stevioside has been widely used in the production of food, beverages and seasonings in Asia, North America, South America and the European Union. China is the world's leading producer of stevia.

Stevia has the characteristics of high sweetness and low heat energy. Its sweetness is 200-300 times that of sucrose, and the calorific value is only 1/300 of sucrose. It has been proved by a large number of scientific experiments that stevioside A glycosides are non-toxic and have no side effects, and are an ideal sweetener for replacing sucrose. In addition, stevioside can be widely used in food, beverage, seasoning, brewing, medicine and other industries.

The stability and metabolic pathways of stevioside have been studied intensively, and the safety of high-purity stevioside has been studied intensively. The crude stevioside extract has been used as a food additive since the mid-1990s. In 2008, pure stevioside A was first recognized by the US Food and Drug Administration as "GRAS (often considered safe)".

Stevia form 1, form 2, form 3A, form 3B, amorphous form and preparation thereof are reported in the patent US 20070292582 - A1; Form 1, Form 2 and Solubility are reported in the patent WO20101 18218A1. Very high crystal form 3 and its preparation method, wherein the crystal form 2 is the same as the crystal form 1 reported in the patent US 20070292582 - A1, and the crystal form after the crystal form 1 is dried is the same as the US 20070292582 - A 1 crystal form 3A, 3B . In addition, an article entitled "Single Crystal Growth and Structure Determination of the Natural " High Potency " Sweetener Rebaudioside A" in Crystal Growth & Design magazine reported the methanol tetrahydrate Form III. , the crystal form is unstable. Form 1 in the patent US 20070292582 - A 1 corresponding to Form I in this article, Form II corresponds to Form 3A, Form 3B in US 20070292582 - A 1 , and Form IV corresponds to US 20070292582 - A1 Form 2.

There is an urgent need in the art to provide a crystal form having better properties, such as a new crystal form having high crystallinity, good solubility, and high stability. Summary of the invention

The present invention aims to provide a novel stevioside A glycoside crystal.

Another object of the present invention is to provide a process for the preparation of the novel stevioside A glycoside crystal.

A further object of the invention is to provide the use of said novel stevioside A glycoside crystals. In a first aspect of the invention, there is provided a Stevia A glycoform, wherein the structure is as shown in Formula I, The X-ray powder diffraction (XRPD) pattern of the Form 9 has characteristic peaks at the following 2Θ±0.1° angles: 4.748, 9.511, 11.767, 13 23.428;

In another preferred embodiment, the X-ray powder diffraction (XRPD) pattern of Form 9 has characteristic peaks at the following 2 Θ ± 0.1° angles: 4.86, 6.00, 8.82, 9.02, 9.64, 11.90, 13.68, 14.13 14.79, 15.86, 16.40, 17.31, 18.06, 18.70, 19.33, 21.27, 21.76, 22.83, 23.47, 25.30, 25.76.

In another preferred embodiment, the crystal form 9 has an X-ray powder diffraction (XRPD) pattern as shown in FIG.

In another preferred embodiment, the crystalline form 9 differential scanning calorimetry has no characteristic endothermic peak at 50-250 °C. In a second aspect of the invention, there is provided a process for the preparation of stevioside A glycoside form 9 provided by the invention as described above, the method comprising the steps of:

(1) mixing stevioside and a solvent at room temperature to obtain a suspension 1;

(2) stirring the suspension 1 to obtain a suspension 2;

(3) The suspension 2 was filtered and dried to obtain a stevioside crystal 9 crystal.

In another preferred embodiment, the solvent in the step (1) is tetrahydrofuran, and/or n-heptane. In another preferred embodiment, the stirring speed in the step (2) is 60-600 rpm; and the stirring time is 2 hours - 2 days.

In another preferred embodiment, in step (3), suspension 2 is filtered, solvent washed and dried to obtain stevioside crystal 9; the solvent is selected from low boiling point ether.

In another preferred embodiment, the drying in the step (3) is dried at 50 ° C or dried under reduced pressure. In a third aspect of the invention, there is provided a use of the stevioside A glycoside form 9 provided by the invention as described above for the preparation of a food or a medicament. Accordingly, the present invention provides a crystal form having better properties, such as a new crystal form having high crystallinity, good solubility, and high stability. DRAWINGS

Fig. 1 is an X-ray powder diffraction (XRPD) pattern of the stevioside A glycoside form 9 obtained in the examples. Fig. 2 is a graph showing the thermogravimetric analysis (TG) of the stevioside A glycoside form 9 obtained in the examples.

Fig. 3 is a differential scanning calorimetry (DSC) chart of the stevioside A glycoform form 9 obtained in the examples. Fig. 4 is a dynamic water vapor adsorption (DVS) diagram of the stevioside A glycoform form 9 obtained in the examples. detailed description

After several attempts, the inventors discovered a new stevioside A glycoform, crystal form 9, and found a simple preparation method for obtaining the crystal form. On the basis of this, the present invention has been completed. As used herein, "compounds of formula I", "compounds of formula 1" or "rebaudioside" are used interchangeably and are all compounds having the structure shown below:

The words "compound", "composition", "agent" or "medicine or medicament" are used interchangeably herein and refer to when applied to a body. (human or animal), the pharmacy that can be induced by local and/or systemic effects And/or a compound or composition that is physiologically reactive.

As used herein, "room temperature" means 15-30 ° C, preferably 20-25 ° C. Identification and properties of stevioside A glycoforms

The inventors further studied the properties of the stevioside A glycoside using a variety of methods and instruments.

"X-ray powder diffraction", also known as "X-ray polycrystalline diffraction (XRPD)", is a common test method currently used to determine crystal structure (ie, crystal form;). An X-ray powder diffractometer is used to generate a series of diffraction patterns when X-rays are transmitted through the crystal. The different diffraction lines and their intensities in the spectrum are determined by the atomic groups of a certain structure, thereby determining the specific crystal structure of the crystal.

Methods for determining X-ray powder diffraction of crystals are known in the art. For example, using a Bmker D8 Advanced model X-ray powder diffractometer, a copper radiation target is used to acquire the spectrum at a scan rate of 2° per minute.

The stevioside A genotype 9 of the present invention has a specific crystal morphology and has a specific characteristic peak in an X-ray powder diffraction (XRPD) pattern. Specifically, the X-ray powder diffraction (XRPD) pattern of the stevioside A glycoform form 9 of the present invention has eight strong peaks at the following 2 Θ ± 0.1° angles: 4.748, 9.511, 11.767, 13.598, 14.038, 17.224, 19.242 , 23.428; There are characteristic peaks at the following 2Θ ± 0.1° angles: 4.80, 5.48, 8.42, 9.27, 11.05, 11.27, 11.86, 12.62, 13.59, 14.20, 15.07, 15.44, 17.05, 17.72, 18.13, 18.62, 19.36, 21.26 21.95, 22.75, 23.59, 24.14, 24.73, 25.01, 25.54, 25.98, 26.56. In a preferred embodiment of the invention, the stevioside form 9 has an X-ray powder diffraction (XRPD) pattern substantially identical to that of Figure 1.

"Differential scanning calorimetry", also known as "differential calorimetric scanning analysis" (DSC), is a technique for measuring the relationship between the energy difference between a test substance and a reference material and temperature during heating. The position, shape and number of peaks on the DSC map are related to the nature of the material and can therefore be used qualitatively to identify the substance. This method is commonly used in the art to detect various parameters such as phase transition temperature, glass transition temperature, and heat of reaction of a substance.

DSC assay methods are known in the art. For example, a DSC scan of the crystal can be obtained by using a DSC Q20 differential scanning calorimeter at a temperature rise rate of 10 °C per minute from 25 °C to 300 °C.

In one embodiment of the present invention, the stevioside A glycoside form 9 differential scanning calorimetry obtained by the method of the present invention is measured by DSC to have no characteristic endothermic peak at 50-250 ° C, preferably the sweetness of the present invention. Inulin A glycoform form 9 has a DSC pattern substantially identical to that of Figure 3.

"Thermal Weightlessness Analysis" (TG) can analyze the moisture, volatile matter, ash, fixed carbon and LOIo of the analyte.

Methods of determining TG are known in the art. For example, a dynamic moisture adsorber can be used.

In one embodiment of the present invention, the stevioside A glycoside form 9 obtained by the method of the present invention has a TG pattern substantially identical to that of Figure 2 as measured by TG.

"Dynamic Water Vapor Adsorption" (DVS) is a "Dynamic Water Vapor Adsorption" (DVS) instrument that measures the hygroscopicity of a sample. Methods for its determination are known in the art. For example, Surface Measurement Systems, Ltd. instruments can be used to collect data from 5% to 95% humidity at 25 degrees Celsius. The weight deviation under each humidity is not more than ± 0.02% within lOmin. .

DVS assay methods are known in the art. For example, a dynamic moisture adsorber can be used. The stevioside A glycoside form 9 of the present invention has a specific stability and is advantageous for preservation. The inventors showed by the DVS spectrum that in the conventional storage environment (40% - 80% RH), Form 9 has no or almost no hygroscopicity. In a preferred embodiment, the DVS pattern of the resulting stevioside A glycoform 9 is substantially identical to that of Figure 4. Stevioside A glycoside form 9 preparation method

The present invention provides a method of preparing the stevioside A form 9, wherein the method comprises the steps of:

The first step is to carry out the suspension, that is, mixing the stevioside and the solvent at room temperature to obtain a suspension 1; the second step is stirring, that is, the suspension 1 obtained in the first step is stirred to obtain a suspension 2 The third step is to obtain a crystal, which is obtained by filtering the suspension 2 obtained in the second step, and then drying to obtain a stevioside crystal 9 crystal.

The solvent involved in the first step is tetrahydrofuran and/or n-heptane. In a mixed solvent of tetrahydrofuran and n-heptane, the volume ratio of tetrahydrofuran to n-heptane is 2: 1-1: 0, preferably pure tetrahydrofuran (1:0).

The stirring speed involved in the second step is preferably 60-600 rpm; the stirring time is preferably 2 hours - 2 days, more preferably 2 hours - 10 hours.

In the third step, the suspension 2 is filtered, and the solid portion is washed with a solvent and then dried. The drying is preferably dried at 50 ° C or dried under reduced pressure. The solvent used for washing in the third step is selected from low boiling point ether. Use of stevioside A glycoform form 9 and composition thereof

The invention further relates to a composition comprising a novel crystalline form of Stevia A glycoside provided by the present invention, said composition comprising an effective amount of Stevia A glycoside Form 9 and a food/pharmaceutically acceptable carrier. As used herein, the term "containing" or "including" includes "comprising", "consisting essentially of", and "consisting of". The term "effective amount" refers to an amount which is functional or active against a human and/or animal and which is acceptable to humans and/or animals.

The term "pharmaceutically acceptable" or "food acceptable" ingredients is suitable for humans and

/ or animals without excessive adverse side effects (such as toxicity, irritability, and allergies;), that is, substances with reasonable benefits/risk ratios.

Preferably, the "pharmaceutically acceptable carrier" is selected from the group consisting of: a filler, a disintegrant, a lubricant, a glidant, an effervescent agent, a flavoring agent, a coating material, an excipient, or a gentle/ Controlled release agent. In the composition, the pharmaceutically acceptable carrier may contain a liquid such as water, saline, glycerol and ethanol. In addition, auxiliary substances such as fillers, disintegrants, lubricants, glidants, effervescent agents, wetting or emulsifying agents, flavoring agents, pH buffering substances and the like may also be present in these carriers. Generally, these materials can be formulated in a non-toxic, inert, and pharmaceutically acceptable aqueous carrier medium wherein the pH is usually from about 5 to about 8, preferably, the pH is from about 6 to about 8. The above-mentioned features mentioned in the present invention, or the features mentioned in the embodiments, may be arbitrarily combined. All of the features disclosed in the present specification can be used in combination with any of the compositions, and the various features disclosed in the specification can be substituted for any alternative feature that provides the same, equal or similar purpose. Thus, the features disclosed are only general examples of equal or similar features, unless specifically stated. The main advantages of the invention are:

1. The new crystal form provided by the present invention has high crystallinity, remarkable solubility, and a certain degree of improvement in chemical and physical stability.

2. The new crystal form provided by the invention has good chemical and physical stability, and the crystal form formed is regular, which is beneficial to the process treatment of stevioside A glycoside and industrial application.

3. The method for preparing new stevioside A glycoforms provided by the invention is simple and easy to industrialize. Production. The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are merely illustrative of the invention and are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually carried out according to conventional conditions or according to the conditions recommended by the manufacturer. All percentages, ratios, ratios, or parts are by weight unless otherwise indicated.

The unit of weight percent by volume in the present invention is well known to those skilled in the art and, for example, refers to the weight of the solute in a 100 ml solution.

Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be used in the methods of the invention. The preferred embodiments and materials described herein are for illustrative purposes only. Experimental conditions:

XRPD: All XRPD spectra of this patent are detected by the Brooke D8 Advance X-ray diffractometer at room temperature, 2 angstrom scans from 3 degrees to 40 degrees, Cu K , scanning speed: 0. Γ / step.

It should be noted that in the X-ray diffraction pattern of the powder sample, the specific crystal form of the diffraction spectrum obtained from the crystalline compound is often characteristic, and the relative intensity of the band (especially at low angles) may be due to crystallization conditions. The effect of the dominant orientation effect due to the difference in particle size, relative content of the mixture, and other test conditions. Therefore, the relative intensity of the diffraction peaks is not characteristic for the crystals to be targeted. When judging whether they are the same as the known crystal forms, more attention should be paid to the positions of the peaks rather than their relative intensities. In addition, care should be taken to maintain the overall concept when determining whether the crystal form is the same, because not a diffraction line represents a phase, but a specific set of "" data represents a phase. It should also be noted that in the identification of the mixture, some of the diffraction lines are missing due to factors such as a decrease in content. At this time, it is not necessary to rely on all the bands observed in the high-purity sample, and even one band may be given. The crystals are characteristic.

DSC: All DSC spectra of this patent were measured by a DSC 8500 differential scanning calorimeter from Elmer, Platinum, USA, with an atmosphere of nitrogen and a heating rate of 10 degrees Celsius per minute.

DVS: All of the dynamic moisture adsorption (DVS) experimental data of this patent were measured by the British SMS instrument company DVS Intrinsic type dynamic moisture adsorber. Measurement conditions: Temperature: 25 Relative humidity range: 5%-95%. The raw stevioside in the following examples was purchased from Zhucheng Haotian Pharmaceutical Co., Ltd. Example 1

30 mg of stevioside and 1 mL of solvent tetrahydrofuran were suspended at room temperature; the suspension was stirred with a magnetic stirrer at a speed of 60-600 rpm for 2 days; the suspension was vacuum filtered. After washing with the mother liquor, the stevia crystal form 9 is obtained by drying at 50 ° C under normal pressure. Example 2

30 mg of stevioside and 1 mL of solvent tetrahydrofuran were suspended at room temperature; the suspension was stirred with a magnetic stirrer at a speed of 60-600 rpm for 2 days; the suspension was vacuum filtered. After washing with the mother liquor, it was dried under reduced pressure at 50 ° C to obtain Stevia Form 9. Example 3

30 mg of stevioside and 1 mL (volume ratio of 1:1) of solvent tetrahydrofuran and n-heptane were mixed at room temperature; the suspension was stirred with a magnetic stirrer at a speed of 60-600 rpm, and the stirring time was 2 days; The above suspension was vacuum filtered, washed with a solvent, and dried at 50 ° C under normal pressure or reduced pressure to obtain Stevia Form 9. Example 4

30 mg of stevioside and 1 mL of solvent tetrahydrofuran were suspended at room temperature; the suspension was stirred with a magnetic stirrer at a speed of 60-600 rpm for 2 hours; the suspension was vacuum filtered. After washing with the mother liquor, the stevia crystal form 9 is obtained by drying at 50 ° C under normal pressure. The maximum solubility of Form 9 obtained in the above examples was about 80 mg/mL, and the crystal form which was left at room temperature for 3 months was not detected by XRPD. Purity (purity by high performance liquid phase method, using the same liquid column and liquid phase method as specified by JECFA in 2010) varies by less than 0.5%.

Preparation of sample solution: Accurately weigh 50-100 mg of stevioside sample, put it into a 50 ml volumetric flask, and then add 7:3 water acetonitrile solution to dissolve to 50 ml mark. Detection procedure: 5 μl of the sample solution was injected under the following conditions. Column: Shiseido's Capcell pak C 18 MG II column or Phenomenex's Luna 5 μ CI 8(2) 100A column or equivalent column (length: 250 mm; inner diameter: 4.6 mm, packing) Particle size: 5 μ ηι). Flow Phase: A mixture of acetonitrile and sodium phosphate buffer (specification: 10 mmol/L, pH 2.6) in a ratio of 32:68. How to configure the sodium phosphate buffer: 2.76 g of sodium dihydrogen phosphate was dissolved in 2 liters of water, and the pH was adjusted to 2.6 by adding phosphoric acid: 1 ml/min. Detector: 210 nm UV detection. Column temperature: A trace of approximately 30 minutes was recorded at 40 °C. The above is only the preferred embodiment of the present invention, and is not intended to limit the scope of the technical scope of the present invention. The technical content of the present invention is broadly defined in the scope of the claims of the application, any technical entity completed by others. The method or method, if it is identical to the scope of the claims, or equivalents, is considered to be within the scope of the claims.

Claims

Wo

A stevioside glycoside crystal form 9 having the structure shown in Formula I, wherein the X-ray powder diffraction (XRPD) pattern of the Form 9 has characteristic peaks at the following 2 Θ ± 0.1 ° angle: 4.748, 9.511 , 11.767, 13.598, 14.038, 17.224, 1

2. The stevioside form 9 according to claim 1, wherein the crystal form 9 has an X-ray powder diffraction (XRPD) pattern having a characteristic peak at the following 2 Θ ± 0.1 ° angle: 4.86 6.00, 8.82, 9.02, 9.64, 11.90, 13.68, 14.13, 14.79, 15.86, 16.40, 17.31, 18.06, 18.70, 19.33, 21.27, 21.76, 22.83, 23.47, 25.30, 25.76.

The stevioside form 9, according to claim 1, wherein the crystal form 9 has an X-ray powder diffraction (XRPD) pattern as shown in Fig. 1.

The stevioside A glycoside form 9, according to claim 1, wherein the crystal form 9 differential scanning calorimetry has no characteristic endothermic peak at 50 to 250 °C.

5. A method of producing stevioside A glycoforms 9 according to any one of claims 1 to 4, wherein the method comprises the steps of:

(2) stirring the suspension 1 to obtain a suspension 2; (3) The suspension 2 was filtered and dried to obtain a stevioside crystal 9 crystal.

The process according to claim 5, wherein the solvent in the step (1) is tetrahydrofuran, and/or n-heptane.

The preparation method according to claim 5, wherein the stirring speed in the step (2) is 60-600 rpm; and the stirring time is 2 hours - 2 days.

The preparation method according to claim 5, wherein in the step (3), the suspension 2 is filtered, the solvent is washed, and then dried to obtain a stevioside crystal 9 crystal; and the solvent is selected from a low boiling point ether.

The preparation method according to claim 5 or 8, wherein the drying in the step (3) is dried at 50 ° C or dried under reduced pressure. 10. Use of a stevioside A glycoformine form 9 according to any one of claims 1 to 4 for the preparation of a food or a medicament.