KR20180050451A - Preparation Method for Crystalline L-α-Glyceryl Phosphoryl Choline - Google Patents

Abstract

The present invention relates to a new preparation method of a L-α-glyceryl phosphoryl choline crystal, the preparation method which is capable of preparing the crystal by a method of facilitating preparations of the crystal and facilitating mass production of the crystal due to low hygroscopic properties of the crystal. More specifically, the present invention relates to a preparation method of a L-α-glyceryl phosphoryl choline crystal, the preparation method comprising the steps of: a moisture content adjusting step (A) of concentrating a liquid L-α-glyceryl phosphoryl choline to adjust a moisture content of the L-α-glyceryl phosphoryl choline to 4 to 8 wt%; an alcohol dissolution step (B) of injecting a C_1-C_3 alcohol into the L-α-glyceryl phosphoryl choline, having moisture content thereof adjusted in the step (A), in an amount (v/w) of 1 to 3 times corresponding to that of the L-α-glyceryl phosphoryl choline to dissolve the L-α-glyceryl phosphoryl choline at 45 to 55°C to obtain a solution; an acetone injection step (C) of injecting acetone into the solution of the step (B) in an amount (v/v) of 1.5 to 4 times corresponding to that of the alcohol, and additionally stirring the solution and acetone at 45 to 55°C for 0.1 to 1 hour to obtain a solution; and a crystal obtaining step (D) of cooling the solution of the step (C) to 5 to 15°C, stirring the cooled solution for 1 to 10 hours to precipitate a crystal, filtering the crystal, and drying the filtered crystal.

Description

Preparation Method for Crystalline L-α-Glyceryl Phosphoryl Choline]

The present invention relates to a process for producing L-α-glycerylphosphoryl choline crystals which is easy to apply in mass production, has high drying efficiency and low hygroscopicity of the crystals thus prepared, and is suitable for formulation.

L-α-glycerylphosphorylcholine (also referred to as L-α-Glyceryl Phosphoryl Choline or choline alfoscerate) is a choline compound present in the brain.

[Chemical Formula 1]

L-α-glycerylphosphorylcholine is a precursor of acetylcholine, a stimulant of parasympathetic nerves. It plays a role in rapidly transporting choline to the brain through the blood-brain barrier, thus normalizing cranial nerve cells and the cholinergic system. In addition, since it acts directly on the brain, it has excellent effects and safety, and is widely used as a non-prescription drug in many countries, including the United States, as an agent for treating Alzheimer's disease and dementia and improving brain function.

Since L-α-glycerylphosphoryl choline has a very high hygroscopicity and easily absorbs moisture in the air, L-α-glycerylphosphoryl choline, which is conventionally produced and sold in the industry, is a liquid phase containing water, Commercially available formulations are also limited to soft capsule formulations using liquid L-a-glycerylphosphoryl choline. Since the content of the liquid raw material is fluid, it is not easy to accurately measure the effective amount, and there is a risk of being altered by microorganisms, and a preservative should be used and it is difficult to apply it to various formulations. In addition, soft gelatin capsules are disadvantageous in that the gelatin capsules can be dissolved by moisture contained in the soft gelatin capsules over time.

Therefore, researches on crystallization methods that can be used for various formulations and easy to circulate and store have been continued. U.S. Patent No. 5,523,450 discloses a process for producing an L-α-glycerylphosphorylcholine starting material by concentrating a liquid L-α-glycerylphosphoryl choline raw material to reduce the water content to 4% or less and then crystallizing it in ethanol to form an alcoholate crystal . However, the crystals obtained by the above method need to be packaged in an airtight container in the presence of argon, so that it can be deduced that the hygroscopicity is still very high. If the hygroscopicity is large, the weight and physical properties may change during the storage or formulation of the raw material, and if the moisture content increases, it may permeate into the packaging material or adhere to the packaging material and cause loss.

Hanse Chem Co., Ltd. is a crystalline polymorph of the prior art. It is a crystalline form of L-α-glycerylphosphorylcholine by crystallization using ethanol or isopropanol as a solvent, depending on the water content before crystallization of the liquid raw material and stirring, (I-form, II-form and III-form) can be obtained, respectively (Patent No. 10-1287422, 10-1287424, 10-1287423, 10-1287425, and 10-2015-0086160). Of these, type I is anhydride, type II is monohydrate, and type III is monohydrate. The pharmacologically meaningful crystalline form should have a melting point of at least 100 캜, but the II-type crystal has a melting point of 66 캜, which is practically useless for formulation. The conditions for the I and III crystallization are the difference in the water content (6-12% vs 3-5%) of the liquid L-α-glycerylphosphoryl choline, and when the seed crystal is not seeded, It takes more than 24 hours. The I-form crystals and II-form crystals produced by the above method were delayed in humidification rate as compared with the conventional polymorphic crystals because the polymorphic crystals prepared by the conventional method had a rough crystal surface and a large irregularity. This suggests that the hygroscopicity can be controlled depending on the shape of crystals formed even if polymorphous crystals are formed. Since the I-form crystal according to the present invention still has moisture tolerance, it is necessary to preserve the temperature and humidity in a controlled state in order to preserve seed crystals for several days. In addition, if the water content is not precisely controlled, mixed crystals of I-form and III-form can be formed, and II-form crystals can be formed together when the stirring during crystallization is not sufficient. The crystallization conditions must be precisely controlled.

JT H Biotech Co., Ltd. points out the disadvantage that the Form I crystal requires seeding, and discloses a new crystalline type manufacturing method which is useful for pharmaceuticals in the patent 10-1582302. The method comprises dissolving glycerylphosphorylcholine in a liquid phase in an organic solvent, removing moisture by treating with a desiccant of magnesium sulfate or sodium sulfate, and filtering the mixture in a mixture of ethylacetate and methanol in a volume ratio of 3 to 7: 1 for 6 hours Followed by stirring for 5 days to crystallize. However, since the above method (1) requires a long time for filtration of the hygroscopic agent because the amount of the hygroscopic agent used is 1 to 2 times that of L-? -Glycerylphosphoryl choline, L-? -Glycerylphosphoryl choline remains in the filtered hygroscopic agent (2) If water is contained in the methanol used as a solvent for the desiccant, magnesium sulfate and sodium sulfate may partially dissolve. Therefore, in the crystallization process using a mixture of ethyl acetate and methanol, L- a hygroscopic agent may precipitate together with? -glycerylphosphoryl choline to reduce purity. (3) Since the amount of the absorbent and the solvent used is high, the raw material cost is high. (4) Since the moisture absorbent needs to be treated as waste after the moisture treatment, the waste disposal cost is high and environmental problems are caused. It is not competitive in the field.

United States Patent 5,523,450 Patent No. 10-1287422 Patent No. 10-1287424 Patent No. 10-1287423 Patent No. 10-1287425 Patent Document 10-2015-0086160 Patent No. 10-1582302

Disclosure of Invention Technical Problem [8] To solve the problems of the prior art as described above, it is an object of the present invention to provide a process for crystallizing L-α-glycerylphosphorylcholine by mass production in an industrial field and an economical process.

Another object of the present invention is to provide a method for producing a crystal having a high drying efficiency and a low hygroscopicity as compared with the conventional crystal form.

In order to accomplish the above object, the present invention provides a method for preparing a liquid composition, comprising: (A) controlling a water content to adjust a water content to 4 to 8 w% by concentrating L-α-glyceryl phosphoryl choline in a liquid phase; (B) 1 to 3 times (v / w) of a C1 to C3 alcohol is added to L-? -Glycerylphosphoryl choline in which the water content is controlled in the step (A), and the mixture is dissolved at 45 to 55 占 폚 An alcohol dissolving step; (C) adding acetone 1.5 to 4 times (v / v) of the alcohol to the solution of step (B) and further stirring at 45 to 55 ° C for 0.1 to 1 hour; And (D) cooling the solution of step (C) to 5 to 15 DEG C and stirring for 1 to 10 hours to precipitate crystals, followed by drying. To a process for producing glycerylphosphoryl choline crystals.

Hereinafter, each step will be described in detail.

(A) Moisture content control step

L-α-glycerylphosphorylcholine is usually distributed in a liquid state and contains a large amount of water. Since L-α-glycerylphosphoryl choline has a very high solubility in water, crystallization is difficult as the solvent contains a large amount of water at the time of crystallization, and the yield is low even if crystallization occurs. Therefore, it is necessary to remove moisture from the raw material in advance and adjust the water content to an appropriate level before crystallization.

In this step, the moisture content is adjusted to 4 to 8 wt%. If the moisture content is too high, crystallization is difficult or the yield of crystallization is lowered as described above. If, on the other hand, the moisture content is too low, an excess of alcohol is required to dissolve the L-α-glycerylphosphorylcholine in step (B) below.

In this step, the concentration of the L-α-glycerylphosphorylcholine in the liquid phase is preferably concentrated in vacuo at 45 to 85 ° C. When the temperature at the time of vacuum concentration is too low, the concentration efficiency naturally decreases, so that it takes a long time to concentrate and the process cost increases. On the other hand, if the concentration temperature is too high, the L-α-glycerylphosphoryl choline may be denatured.

At this time, the alcohol of L-alpha -glycerylphosphoryl choline C1-C3 may be added and concentrated together so that the concentration of water can proceed efficiently. The C1 to C3 alcohols are azeotropic with water, allowing water to be removed more quickly at low temperatures. When the alcohol is concentrated by concentration, it is easy for a person skilled in the art to measure the amount of alcohol remaining in the residue and the amount of alcohol to be administered in the step (B).

(B) Alcohol dissolution step

When the moisture content of the L-α-glycerylphosphorylcholine sample is controlled, a 1 to 3-fold (v / w) amount of a C1 to C3 alcohol is added and the L- To dissolve completely.

As the C1 to C3 alcohol, methanol, ethanol, propanol or isopropanol may be used alone, or a mixture of two or more C1 to C3 alcohols may be used. It is most preferred to use a mixture of methanol and isopropanol in a ratio of 1: 1 to 3 (v / v). The alcohol is used in an amount of 1 to 3 (v / w) of L-α-glycerylphosphoryl choline, and when the amount of alcohol is too small, L-α-glycerylphosphoryl choline does not sufficiently dissolve, If the amount of alcohol is too large, the amount of acetone used in step (C) may be too large and the yield may be lowered.

If the temperature is too low, the dissolution may not be sufficient. If the temperature is too high, it is difficult to carry out the operation at the time of introducing acetone in the step (C) below.

(C) Acetone feeding step

In this step, acetone of 1.5 to 4 times (v / v) of alcohol is added to the alcohol solution of step (B) and further stirred at 45 to 55 ° C for 0.1 to 1 hour to prepare a homogeneous solution. When the amount of acetone used is too small, the yield decreases. When the amount of acetone used is too large, amorphous crystals may precipitate rapidly or the uniformity of crystals may be deteriorated. If the stirring time is too short, time is not enough to form a homogeneous solution. Even if the stirring time is long, there is no problem, but about one hour is sufficient, so stirring for a long time is not necessary.

In this step, it is more preferable to maintain the temperature of the acetone introduction process at 45 to 55 ° C. If the temperature is too low at the time of charging, amorphous crystals may precipitate or coalesce during the charging process. Considering that the boiling point of acetone is 56.5 캜, it is preferable to keep the temperature at 55 캜 or lower during the addition.

(D) Crystallization step

In this step, the solution of step (C) is cooled to 5 to 15 ° C, stirred for 1 to 10 hours, and the precipitated crystals are filtered and dried.

This step should be stirred at 5 to 15 ° C for at least one hour so that crystals can be precipitated and grown. If the agitation time is too short, crystal precipitation and growth may not be sufficient and the yield may be lowered. Even if the agitation time exceeds 10 hours, it is not a problem, but it is more preferable to stir the crystal precipitation step for 1 to 5 hours in consideration of economical efficiency of the process. In this step, seeding may be performed to promote crystal precipitation and growth, but this is not essential. The stirring speed was preferably 30 to 300 rpm.

The filtered crystals were dried in vacuo to give the final L-α-glycerylphosphoryl choline crystals. According to the crystallization method of the present invention, crystalline L-α-glycerylphosphoryl choline can be prepared from a liquid L-α-glycerylphosphoryl choline starting material at a high yield of 85 to 95% based on the dry weight.

The crystals of L-α-glycerylphosphoryl choline produced by the method of the present invention showed a sharp diffraction pattern in a powder X-ray diffraction analysis to show a crystalline form. However, depending on the production batch or even in the same batch, the relative intensity of the diffraction peaks varied with sampling. Fig. 1 shows the X-ray diffraction spectra of the crystals obtained by the method of the present invention. The diffraction pattern shown near 2? = 20 占 consists of two separated peaks. According to the samples 1) in the above spectrum, In the middle spectrum was 0.85: 1, and in the lower spectrum was 1.8: 1 in the middle spectrum. This suggests that the crystals obtained by the method of the present invention are polymorphic crystals consisting of a mixture of at least two crystal forms.

The crystals according to the present invention are anhydrous and have a high drying efficiency, so that the content of water in the final crystals is lower than that of conventional single crystals. That is, the I-form crystals prepared in Comparative Example had no significant difference in moisture content even after drying at 55 ° C. for 12 hours and then further drying for 2 hours at a moisture content of 0.16%. However, The dried crystals showed moisture content of 0.04 ~ 0.08% as a result of batch drying after drying at 55 ℃ for 9 hours, indicating that the drying efficiency was excellent. This is a very low value compared to the initial moisture content of the monoclinic forms described in the published patent 10-2015-0086160. Since the initial moisture content also affects the rate of humidification, it is another advantage over the prior art that crystals with low initial water content can be produced.

In addition, the crystals obtained by the present invention have a hygroscopicity that is significantly lower than that of the conventional crystals even though they are polymorphic crystals. This means that crystals suitable for formulation can be prepared according to the crystallization method even if they are not single crystals.

As described above, according to the method of producing the crystalline form of L-α-glycerylphosphoryl choline of the present invention, since moisture is not removed by using the moisture absorbent, there is no loss in the filtration process of the moisture absorbent treatment, The process is not only performed by a simple process of concentration of water, solvent, and cooling, but also is advantageous for mass production because crystallization takes place in a short time even without seeding seed seed crystals.

The crystals of L-? -Glycerylphosphoryl choline produced by the method of the present invention have a higher drying efficiency than that of a single crystal type known by the prior art, so that when dried under the same conditions, the water content is much lower than that of the conventional single crystal form, Since the humidification rate is also slow, the storage stability is high and the handling is easy, so that it can be formulated into various forms.

1 is an X-ray diffraction spectrum of crystals of L-α-glycerylphosphoryl choline of the present invention.
Fig. 2 is a differential scanning calorimetry spectrum of crystals of L-? -Glycerylphosphoryl choline of the present invention. Fig.
3 is a graph showing the hygroscopicity of crystals of L-α-glycerylphosphorylcholine prepared by the present invention and the prior art.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these embodiments are merely examples for explaining the content and scope of the technical idea of the present invention, and thus the technical scope of the present invention is not limited or changed. It will be apparent to those skilled in the art that various changes and modifications can be made within the scope of the technical idea of the present invention based on these examples.

Example: Preparation of L-? -Glycerylphosphoryl choline crystal

460 g of liquid L-a-glycerylphosphoryl choline (moisture content 15 w%) was concentrated in vacuo at 80 캜 for 9 hours to reduce the water content to about 5 w%. 200 ml of MeOH and 400 ml of IPA were added thereto and dissolved with stirring while maintaining the temperature of 50 ± 5 ° C. 1400 ml of Acetone was then added, and further stirred for 30 minutes while maintaining the temperature at 50 ± 5 ° C .

The L-α-glycerylphosphoryl choline solution was then cooled to 9 ° C. and stirred at 200 rpm for 2 hours. The precipitated crystals were filtered and vacuum-dried at 55 ° C for 9 hours to obtain 350 g (yield: 89.5%) of L-α-glycerylphosphoryl choline crystals having a moisture content of 0.08%.

Comparative Example: Preparation of I-form crystal according to the registered patent 10-1287424

460 g of liquid L-α-glycerylphosphorylcholine (moisture content 15 w%) was concentrated in vacuo at 50 ° C. for 9 hours to reduce the water content to about 7%. 920 ml of ethanol was added thereto, and the mixture was sufficiently dissolved by stirring while maintaining the temperature at 50 캜, and then filtered with a 1.0 탆 membrane filter to remove foreign matter.

Next, the L-α-glycerylphosphorylcholine choline solution was cooled to 9 ° C. and added with L-α-glycerylphosphorylcholine I-form crystals (as described in Patent Document 10-1287434 as seed crystals at -5 ° C. (Hereinafter referred to as " crystallization "), and aged for 2 hours with stirring at a speed of 50 rpm. The precipitated crystals were filtered and vacuum-dried at 55 ° C for 12 hours to obtain 0.31 g (yield: 80%) of L-α-glycerylphosphoryl choline I-form crystal having a moisture content of 0.16%.

Test example: Crystal analysis and physical property test

1. Powder X-ray diffraction (XRD) analysis

Powder X-ray diffraction (XRD) analysis was performed on the L-? -Glycerylphosphoryl choline crystals obtained in the above Examples, and the results are shown in FIG. The measurement conditions of the powder X-ray diffraction (XRD) spectrum are as follows.

1) Device: Ultima IV / X source: Cu

2) Tube voltage: 40 kV / tube current: 40 mA

3) diverging slit: 2/3 ° / scattering slit: 2/3 ° / receiving slit: 0.15 mm

4) Scanning range: 2 to 60 ° 2? / Sampling interval: 0.02 °

5) Scan speed: 3 ° / min

2. Differential scanning calorimetry (DSC) analysis

As a result of the differential scanning calorimetry (DSC) analysis of the crystals obtained in the above examples, as shown in Fig. 2, the endothermic peak was shown at 143 ° C and the onset temperature at 147 ° C. The conditions of the differential scanning calorimetry are as follows.

1) Device: TA Instruments DSC 2910

2) Measuring range: 25 to 200 DEG C / temperature rise interval: 1 DEG C / min

3) N2 rate: 50 ml / min

3. Hygroscopicity evaluation

The hygroscopicity of the L-? -Glycerylphosphoryl choline crystal prepared in the above Example was evaluated.

In the test, the crystal sample of the example was placed in a chamber having a relative humidity of 30% at room temperature (25 ° C), and water content was checked at each time point. The results are summarized in Table 1 below. For comparison, the hygroscopicity of each crystal type at 30% relative humidity described in Japanese Patent Laid-open No. 10-2015-0086160 is shown in Table 1 below. Moisture was measured by Kahl Fisher.

The I-form crystals, the III-form crystals and the IV-form crystals in Table 1 were obtained by vacuum-drying samples at 55 ° C for 12 to 24 hours after crystallization as described in Patent Document 10-2015-0086160, Which was similar to the sample after the production in the above comparative example. That is, although the crystals produced by the examples have a much shorter drying time, the initial moisture content is only about half that of the crystals of Form I, III and IV according to the conventional method, and the moisture absorption rate is also delayed It took a much longer time to reach the reference humidity of 1% of the solid powder of the active active ingredient (API). FIG. 3 is a graph showing the moisture content with time, and it can be seen that the rate of humidification of crystals according to the present invention is significantly lower than that of single crystals I, III and IV.

Claims (6)

(A) a water content regulating step of concentrating L-α-glycerylphosphorylcholine in a liquid phase to adjust the water content to 4 to 8 w%;
(B) 1 to 3 times (v / w) of a C1 to C3 alcohol is added to L-? -Glycerylphosphoryl choline in which the water content is controlled in the step (A), and the mixture is dissolved at 45 to 55 占 폚 An alcohol dissolving step;
(C) adding acetone 1.5 to 4 times (v / v) of the alcohol to the solution of step (B) and further stirring at 45 to 55 ° C for 0.1 to 1 hour; And
(D) cooling the solution of step (C) to 5 to 15 占 폚 and stirring for 1 to 10 hours to precipitate crystals, and then drying the precipitated crystals;
Wherein the L-? -Glycerylphosphoryl choline crystal is prepared by reacting a?
The method according to claim 1,
Wherein the temperature during the step (A) is 45 to 85 占 폚.
The method according to claim 1,
The method for producing L-? -Glycerylphosphoryl choline crystals according to (A), wherein C1-C3 alcohol is added and concentrated together.
The method according to claim 1,
Wherein the alcohol of step (B) is a mixture of methanol and isopropanol in a ratio of 1: 1 to 3 (v / v).
The method according to claim 1,
Wherein the step (B) further comprises, between step (B) and step (C), filtering the solution of step (B) to remove solid matter of solid matter. Gt;
The method according to claim 1,
Wherein the temperature of the step of introducing acetone is maintained at 45 to 55 ° C in the step (C).

Images