CN108017668B

CN108017668B - High-crystal-purity creatine phosphate sodium tetrahydrate hemihydrate and preparation method and application thereof

Info

Publication number: CN108017668B
Application number: CN201610958782.9A
Authority: CN
Inventors: 曾佳烽; 施小风; 李宝莹; 毛杰
Original assignee: Harbin Laiboten Pharmaceutical Co ltd
Current assignee: HARBIN LAIBOTEN PHARMACEUTICAL Co.,Ltd.
Priority date: 2016-11-04
Filing date: 2016-11-04
Publication date: 2020-06-09
Anticipated expiration: 2036-11-04
Also published as: CN108017668A

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a high-crystal-purity creatine phosphate sodium tetrahydrate semi-hydrate and a preparation method thereof, wherein the creatine phosphate sodium crystalline hydrate contains 4.5 water molecules. In the X-ray powder diffraction pattern of the high-crystal-purity sodium creatine phosphate tetrahydrate, the 2 theta angle has characteristic diffraction peaks at 7.6 +/-0.2 degrees, 14.5 +/-0.2 degrees, 15.5 +/-0.2 degrees, 17.3 +/-0.2 degrees, 18.6 +/-0.2 degrees, 21.7 +/-0.2 degrees, 22.8 +/-0.2 degrees and 24.7 +/-0.2 degrees, and the crystal forms of the sodium creatine phosphate 1.5 hydrate and the sodium creatine phosphate 6 hydrate are lower than the detection limit of the conventional scanning of an X-ray powder diffractometer; after the high-crystal-form-purity creatine phosphate sodium tetrahydrate hemihydrate is heated at 120 ℃ for 0.5 hour, no diffraction peak is observed in an X-ray powder diffraction pattern, and the high-crystal-form-purity creatine phosphate sodium tetrahydrate is converted into amorphous. The thermal stability of the creatine phosphate tetrahydrate is obviously superior to that of products on the market and reported crystal 4.5 hydrate process samples, the creatine phosphate tetrahydrate brings great convenience to storage, transportation and use, the storage and transportation cost can be greatly reduced, and the safety of clinical application is improved.

Description

High-crystal-purity creatine phosphate sodium tetrahydrate hemihydrate and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a high-crystal-purity creatine phosphate sodium tetrahydrate hemihydrate and a preparation method and application thereof.

Background

Creatine phosphate (creatine phosphate) is an active substance that is naturally present in the human body and plays an important role in the energy metabolism of muscle contraction. It is a chemical energy reserve for cardiac and skeletal muscle and is used for the resynthesis of ATP. The creatine phosphate disodium salt is clinically used for treating myocardial metabolic disorder in an ischemic state by intramuscular injection or intravenous injection, and is added into cardioplegia solution to protect the myocardium during cardiac surgery. The creatine phosphate disodium salt serving as a myocardial protectant medicament is applied to the market at home and abroad for many years, and the creatine phosphate disodium salt is an endogenous substance of body cells and shows good safety through clinical application. Exogenous creatine phosphate disodium salt is also used in athletic activities to protect skeletal and cardiac muscle from intracellular damage and to enhance muscle function recovery. The creatine phosphate disodium salt has the nerve protection effect, and the clinical application shows that the creatine phosphate disodium salt has the effect of improving the nerve function of patients with large-area cerebral infarction.

Because the aqueous solution of creatine phosphate sodium is unstable and is easy to decompose into creatine, creatinine phosphate and the like, the creatine phosphate sodium sterile powder injection is clinically used at present. The sterile powder injection has high requirement on the purity of the product, the medicine is ensured not to be converted and decomposed in the processes of subpackaging, storing and transporting, and impurities generated by the conversion and the decomposition of the medicine can directly bring the risk of clinical application. Therefore, the high thermal stability has important significance for the drugs of powder injection formulations.

Creatine phosphate sodium has various crystal hydrate crystal forms, and crystal 7 hydrate, crystal 6 hydrate, crystal 4.5 hydrate, crystal 1.5 hydrate and the like are reported, and creatine phosphate sodium for injection which is currently on the market is marked as crystal tetrahydrate, and we find that the main component is actually crystal 4.5 hydrate. Crystalline 4.5 hydrate is a pharmaceutical crystalline form of creatine phosphate sodium, other hydrates are currently non-pharmaceutical crystalline forms. Because the thermal stability of the products in the process on the market is poor, the second part of the Chinese pharmacopoeia 2015 edition (1568 page, under the item of creatine phosphate sodium) requires the storage in a cool and dark place (the dark place is not more than 20 ℃). The light shielding can be realized by packaging, and an additional cooling measure is required below 20 ℃, so that high cost and inconvenience are brought to storage, transportation and use, and meanwhile, the safety risk of clinical application is correspondingly increased. Therefore, how to improve the thermal stability of the sodium creatine phosphate marketed sample is of great significance.

Disclosure of Invention

The invention aims to provide a high-crystal-purity creatine phosphate sodium tetrahydrate to solve the problem that a current clinically used creatine phosphate sodium technical product is sensitive to temperature and unstable.

In order to achieve the above purpose, the invention provides the following technical scheme:

according to a first aspect of the present invention, the present invention provides a high crystal purity sodium phosphocreatine tetrahydrate of formula (1), which meets the following conditions I to III:

I. the high-crystal-form-purity creatine phosphate sodium tetrahydrate hemihydrate has a structural formula shown in the formula (1), and 4.5 crystal water are contained in a molecule;

in the X-ray powder diffraction pattern of the high-crystal-purity creatine phosphate sodium tetrahydrate, characteristic diffraction peaks exist at 2 theta angles of 7.6 +/-0.2 degrees, 14.5 +/-0.2 degrees, 15.5 +/-0.2 degrees, 17.3 +/-0.2 degrees, 18.6 +/-0.2 degrees, 21.7 +/-0.2 degrees, 22.8 +/-0.2 degrees and 24.7 +/-0.2 degrees, the diffraction peak at 18.6 +/-0.2 degrees is called as an a peak, and the corresponding 2 theta angle value is called as a; the content of the crystal forms of the sodium phosphocreatine tetrahydrate 1.5 hydrate and the sodium phosphocreatine tetrahydrate in the high-crystal-purity sodium phosphocreatine tetrahydrate hemihydrate is lower than the detection limit of the conventional scanning of an X-ray powder diffractometer, namely, in an X-ray powder diffraction pattern of the high-crystal-purity sodium phosphocreatine tetrahydrate, no diffraction peak exists at the positions of 8.4 +/-0.2 degrees, 12.1 +/-0.2 degrees, 13.6 +/-0.2 degrees, a +0.4 +/-0.05 degrees and 12.7 +/-0.2 degrees of a 2 theta angle;

and III, after the high-crystal-form-purity creatine phosphate sodium tetrahydrate hemihydrate is heated at 120 ℃ for 0.5 hour, no diffraction peak is observed in an X-ray powder diffraction pattern, and the high-crystal-form-purity creatine phosphate sodium tetrahydrate is converted into amorphous.

The conventional scanning of the X-ray powder diffractometer described in the condition II means that the scanning is performed at a scanning speed of 1.2 to 12 DEG/min.

The high-crystal-purity creatine phosphate sodium crystal tetrahydrate semi-complex is high in purity, the contents of a 1.5 hydrate crystal form and a 6 hydrate crystal form in the high-crystal-purity creatine phosphate sodium crystal tetrahydrate semi-complex are extremely low and are lower than the detection limit of conventional scanning of an X-ray powder diffractometer, namely, characteristic diffraction peaks of other crystal forms cannot be observed in an X-ray powder diffraction pattern of the high-crystal-purity creatine phosphate sodium crystal semi-complex.

JOURNAL OF PHARMACEUTICAL SCIENCES 2014, 103(11): 3688-3695 (hereinafter, referred to as literature 1) reported that different crystalline hydrates OF creatine phosphate have significant differences in X-ray powder diffraction patterns, and the 4.5 hydrates have characteristic diffraction peaks at the 2 theta angles OF 7.6 +/-0.2 degrees, 14.5 +/-0.2 degrees, 15.5 +/-0.2 degrees, 17.3 +/-0.2 degrees, 18.6 +/-0.2 degrees, 21.7 +/-0.2 degrees, 22.8 +/-0.2 degrees, 24.7 +/-0.2 degrees and the like (the diffraction peak at 18.6 +/-0.2 degrees is referred to as an a peak, and the corresponding 2 theta angle value is referred to as a). The crystal form of creatine phosphate sodium crystal 1.5 hydrate has characteristic diffraction peaks at 8.4 +/-0.2 degrees, 12.1 +/-0.2 degrees, 13.6 +/-0.2 degrees, 17.4 +/-0.2 degrees, a +0.4 +/-0.05 degrees and the like (wherein the stronger diffraction peak at 17.4 +/-0.2 degrees is partially overlapped with a weak diffraction peak at the position of the crystal form of 4.5 hydrate). The crystal form of the creatine phosphate sodium crystal 6 hydrate has characteristic diffraction peaks at 12.7 +/-0.2 degrees, a +0.4 +/-0.05 degrees and the like.

The sample of creatine phosphate on the market is marked as crystal tetrahydrate, we find that the crystal is actually 4.5 hydrate with low crystal purity, mixed with 1.5 hydrate crystal form with significant content, the X-ray powder diffraction pattern of the crystal is shown in figures 2 and 3, the characteristic diffraction peak containing 1.5 hydrate crystal form can be obviously observed, and the 2 theta angle in figure 2 is as follows: diffraction peaks at 8.443 °, 12.148 °, 13.632 °, 19.023 °; the 2 θ angle in fig. 3 is: diffraction peaks at 8.386 °, 12.123 °, 13.564 °, 18.984 °.

The high-crystal-purity creatine phosphate sodium tetrahydrate semi-complex prepared by the invention is heated at 120 ℃ for 0.5 hour, is cooled to room temperature, is subjected to X-ray powder diffraction test, and does not have an obvious diffraction peak (shown in figure 7) observed in an obtained spectrum, namely is converted into amorphous. And a sample on the market of creatine phosphate sodium or a creatine phosphate sodium crystal 4.5 hydrate sample reported in literature 1 is heated at 120 ℃ for 0.5 hour, cooled to room temperature, and subjected to an X-ray powder diffraction test, and in an obtained spectrum, a characteristic peak of creatine phosphate sodium crystal 4.5 hydrate disappears, and a characteristic peak of a 1.5 hydrate crystal form is displayed. Indicating a transition to the crystalline 1.5 hydrate form (shown in fig. 8 and 9).

Both document 1 and patent CN104109171A show that creatine phosphate sodium crystal 1.5 hydrate is the most stable crystal form in creatine phosphate sodium hydrate, and as clinical application thereof is not verified, it cannot be used as a pharmaceutical currently. We have unexpectedly found in our studies that for the pharmaceutical crystalline form of creatine phosphate sodium crystalline 4.5 hydrate, the high crystalline purity creatine phosphate sodium tetrahydrate has significantly superior thermal stability compared to the low crystalline purity of the marketed sample mixed with crystalline 1.5 hydrate. That is, for crystalline 1.5 hydrate having high thermal stability, if intermixed in a sample having a crystalline form of crystalline 4.5 hydrate, unexpectedly reduced the thermal stability of the crystalline 4.5 hydrate sample as a whole.

We have made intensive studies and surprisingly found that a sample with high form purity and a sample with low form purity mixed with the 1.5 hydrate form have completely different mechanisms when crystal water is lost by heating. The samples on the market and the 4.5 hydrate process sample described in the literature 1 easily lose part of crystal water at a slightly high temperature, are firstly transformed into a 1.5 hydrate crystal form, and then further lose water at a high temperature. The high crystalline purity creatine phosphate tetrahydrate disclosed in this patent, however, requires a higher temperature at which the water of crystallization is lost and does not first become 1.5 hydrate as in the commercial product, but rather rapidly loses more of the water of crystallization at higher temperatures and becomes amorphous (see example 5).

Thermogravimetric analysis (TG) of the high-crystal-purity creatine phosphate tetrahydrate disclosed by the patent shows that the weight loss is higher in the first stage and lower in the second stage, and the weight loss is generally less than 5.0% at the temperature of between about 150 and 240 ℃. And the creatine phosphate sodium crystal 4.5 hydrate with low crystal form purity, including sodium creatine phosphate marketed samples and creatine phosphate sodium crystal 4.5 hydrate samples reported in literature 1, only loses part of crystal water in the first weight loss stage and is converted into 1.5 hydrate crystal form, and then loses the crystal water in the part of crystal form of 1.5 hydrate in the second stage, and the weight loss between about 150 ℃ and 240 ℃ is usually more than 5.0%.

The sodium creatine phosphate marketed sample is powder injection and is packaged by adopting a closed glass bottle. When the environmental temperature is increased, if the thermal stability is poor, the crystal water is easy to lose, the water which is free in a closed container cannot escape, a solution (pH 8-9) is formed in a microscopic part, and the creatine phosphate sodium is very unstable in an aqueous solution with the pH below 9, so that part of creatine phosphate sodium is rapidly decomposed. As shown in example 8, when the creatine phosphate sodium crystal with high crystal purity is placed at 70 ℃ for 2 hours, the creatine phosphate sodium crystal 4.5 hydrate with high crystal purity has the advantages of unchanged appearance, good fluidity and little increase of related substances; on the other hand, the appearance of the samples on the market and the technical samples of the crystallized 4.5 hydrate disclosed in the literature 1 is seriously caked, and the content of related substances is obviously increased. The high-crystal-purity creatine phosphate sodium tetrahydrate semi-compound disclosed in the patent has excellent thermal stability compared with a sample on the market and a technical sample of a crystalline 4.5 hydrate disclosed in literature 1. The transportation and the storage in high-temperature weather do not need special low-temperature environmental treatment.

It has been unexpectedly found that when a high crystalline purity creatine phosphate tetrahydrate semicomplex of the present invention is mixed with a small amount of crystalline creatine phosphate sodium 1.5 hydrate, even if the content of crystalline 1.5 hydrate is only 0.05% (by mass), the physical properties of the whole substance are greatly changed and the thermal stability is remarkably reduced (examples 10 and 11). After heating at 120 ℃ for 0.5 hour, the X-ray powder diffraction pattern shows the characteristic diffraction peak of the crystalline 1.5 hydrate, which shows a greater weight loss (greater than 5.0%) between about 150 ℃ and 240 ℃ in the second phase of weight loss in thermogravimetric analysis (TG). The properties thereof became consistent with those of the commercially available samples and the 4.5 hydrate sample described in document 1, and the thermal stability was also significantly reduced. The 4.5 hydrate sample mixed with 0.05% crystalline 1.5 hydrate does not observe a characteristic diffraction peak of crystalline 1.5 hydrate crystal form on an X-ray powder diffraction pattern even if the scanning speed is set to 1.2 °/min, because the sensitivity of the X-ray powder diffraction method is low (page 56 of the fourth part of chinese pharmacopoeia 2015 edition, "recognizable in the diffraction pattern when the content of impurity components is greater than 1%).

From the results, it is further shown that the crystal form purity of the creatine phosphate sodium crystal 4.5 hydrate has an extremely significant influence on the thermal stability, and meanwhile, the crystal form purity of the creatine phosphate sodium crystal 4.5 hydrate with high crystal form purity cannot be judged only from an X-ray powder diffraction pattern. At the same time, no better method is available for accurately determining the purity of the crystal form. As discussed above, the creatine phosphate sodium crystalline 4.5 hydrate of high crystalline purity can be concisely characterized by the following two conditions: the characteristic diffraction peak of the 4.5 hydrate crystal form is only shown in an X-ray powder diffraction pattern, and meanwhile, after the sample is heated at 120 ℃ for 0.5 hour, the X-ray powder diffraction measurement shows that the sample is amorphous.

Basically, the samples on the market are mixed with the crystalline 1.5 hydrate with the content higher than the detection limit of X-ray powder diffraction, and the samples are easily judged from the X-ray powder diffraction diagram. On the other hand, the sample of the creatine phosphate sodium crystal 4.5 hydrate described in the literature 1 can not see the characteristic diffraction peak of the 1.5 hydrate crystal form on the X-ray powder diffraction diagram according to the data provided by the literature 1; thermogravimetric analysis shows that the dehydrated weight loss in the first stage is changed into crystalline 1.5 hydrate; heating in an oven at 90 ℃ for 0.5 hour also changed to crystalline 1.5 hydrate. From the results of the crystal form mixing experiments (examples 10 and 11) carried out in this patent, it can be concluded that it is mixed with a small amount of crystalline 1.5 hydrate, and that the crystalline 1.5 hydrate content is below the detection limit of X-ray powder diffraction.

This patent a high crystal form purity creatine phosphate sodium tetrahydrate, its drying process is very easily controlled, dries 1 ~ 2 hours about 50 ℃, and weight is reached invariably promptly, prolongs drying time weight and no longer reduces, surveys moisture with karl fischer method, coincide with the theoretical value. And the water content of the listed sample is controlled to be 20.0-25.0% in the second part of the year 2015 version of Chinese pharmacopoeia (page 1568, under the item of creatine phosphate sodium), the drying end point of the listed sample is not easy to judge in the drying process, and the water content of the product needs to be repeatedly detected in the drying process until the water content reaches 20.0-25.0%. The reason is that as the drying time was extended, the moisture content was continuously decreased until it became 1.5 hydrate, as shown in example 12.

According to another aspect of the present invention, the present invention provides a method for preparing high crystalline purity creatine phosphate sodium tetrahydrate:

1) dissolving creatine phosphate sodium in 1-2 times of water, and stirring to dissolve;

2) controlling the temperature of the system at 0-25 ℃, adding a crystallization agent while stirring, and stirring for 1-5 hours;

3) filtering, washing a filter cake with 95% ethanol or absolute ethanol or acetone, and drying by air flow at 40-60 ℃ to obtain the filter cake;

the crystallization agent is methanol, 95% ethanol or absolute ethanol, and the volume consumption of the crystallization agent is three times or less than three times of the water amount.

Preferably, in the step 1), creatine phosphate sodium is dissolved in 1-1.5 times of water, and is stirred and dissolved.

Preferably, in the step 2), the temperature of the system is controlled to be 0-15 ℃, and the crystallization agent is added under stirring.

Preferably, the crystallization agent is methanol.

Patent CN 103012472 a mentions a crystallization method of creatine phosphate sodium, but the preparation method according to the publication cannot obtain creatine phosphate sodium tetrahydrate with high crystal purity. For example, in example 2 of the present invention, when the volume of 95% ethanol as a crystallization agent is 5 times that of water, the obtained product is a non-high crystal purity creatine phosphate sodium tetrahydrate hemihydrate.

If aseptic operation is adopted, the prepared high-crystal-purity creatine phosphate sodium tetrahydrate hemihydrate is an aseptic product and can be used for preparing aseptic powder injection and can be used as a medicine for protecting cardiac muscle.

The high-crystal-purity sodium creatine phosphate tetrahydrate and sodium creatine phosphate sodium tetrahydrate disclosed by the patent have the obvious difference compared with a crystal 4.5 hydrate sample described in the document 1, have high crystal purity, and have far better thermal stability than the sodium creatine phosphate tetrahydrate and the crystal 4.5 hydrate described in the document 1 (as shown in an example 8 and an example 9).

Although the crystal form of the creatine phosphate crystal 1.5 hydrate has high thermal stability, the crystal form is a non-medicinal crystal form at present, and the clinical application mode, the clinical application way and the clinical application effect of the crystal form are still to be verified, while the high-crystal-purity creatine phosphate tetrahydrate invented by the patent is a clinical application crystal form with higher crystal form purity, can be directly applied to clinic, and can obviously reduce the storage and transportation cost and improve the safety of clinical medication due to the superior thermal stability of the crystal form.

The invention has the beneficial effects that:

1. the crystal form purity of the high-crystal-form-purity creatine phosphate sodium tetrahydrate provided by the invention is far higher than that of a sample on the market. Stable character and easy control of the drying process.

2. The high-crystal-purity creatine phosphate sodium tetrahydrate provided by the invention has thermal stability obviously superior to that of products of the sodium creatine phosphate marketed technology and reported crystallized 4.5 hydrate technical samples, brings great convenience to storage, transportation and use, can greatly reduce the storage and transportation cost, and simultaneously improves the safety of clinical application.

3. The high-crystal-purity creatine phosphate sodium tetrahydrate hemihydrate and the characterization method thereof provided by the invention can effectively distinguish products with low crystal purity, particularly products with low crystal purity, wherein the content of 1.5 hydrate is lower than the detection limit of an X-ray powder diffraction method.

The expression "tetrahydrate" in the present invention is equivalent to "4.5 hydrate", i.e., it means that the crystalline hydrate contains 4.5 water molecules.

The "high-crystal purity creatine phosphate sodium tetrahydrate" prepared by the invention is abbreviated as "crystal hydrate 1" in the examples "

Drawings

FIG. 1: an X-ray powder diffraction pattern of crystalline hydrate 1;

FIG. 2: x-ray powder diffraction pattern of sample 1 on the market made in China;

FIG. 3: x-ray powder diffraction patterns of imported and marketed samples;

FIG. 4: a thermogravimetric analysis (TG) profile of crystalline hydrate 1;

FIG. 5: thermogravimetric analysis (TG) profile of sample 1 made commercially at home;

FIG. 6: thermogravimetric analysis (TG) profile of imported market samples;

FIG. 7: x-ray powder diffraction patterns of the crystalline hydrate 1 after being heated at 90 ℃ and 120 ℃ for 0.5 hour respectively;

FIG. 8: the X-ray powder diffraction pattern of a sample 3 sold in the market made in China after being heated for 0.5 hour at 90 ℃ and 120 ℃ respectively;

FIG. 9: x-ray powder diffraction pattern of sample N (crystalline 1.5 hydrate);

FIG. 10 shows X-ray powder diffraction patterns of a crystalline hydrate 1-mer and a sample-0.05% under a slow scanning condition (step size 0.02 sec, scanning speed 1 sec/step) and X-ray powder diffraction patterns after heating at 120 ℃ for 0.5 hour, respectively.

Detailed Description

The technical solution of the present invention is further described in detail below with reference to specific examples. It should be noted that the following examples are illustrative only and are not to be construed as limiting the scope of the present invention. It will be understood by those skilled in the art that various modifications and equivalent arrangements may be made without departing from the spirit and scope of the present invention and it should be understood that the present invention encompasses the claims appended hereto.

The main equipment used for implementing the patent instrument:

x-ray powder diffractometer, Bruker D8 advance.

Thermogravimetric analyzer, NETZSCH TG 209F 3.

Electric heating constant temperature air-blast drying oven DGG-9076A, Shanghai flowers and Green science instruments, Inc.

Karl Fischer Titrator，METTLER TOLEDO DL38。

A water-isolated constant temperature incubator, model GNP-9080, Shanghai precision testing Equipment Co., Ltd.

The sample 1 on the market made in China is creatine phosphate sodium for injection produced by Jilin Yinji biopharmaceutical corporation, batch number: 20150311.

the domestic sample 2 on the market is creatine phosphate sodium for injection produced by Haikouqi pharmaceutical products Ltd, batch number: 20160401.

sample No. 3, made in China and marketed by Haerbin Ribociton pharmaceutical Co., Ltd, was creatine phosphate sodium salt for injection, lot No. 20160330

The imported samples for marketing are creatine phosphate sodium for injection produced by Alphavisman pharmaceutical company, Italy, batch number: 14868.

the "high-crystalline purity creatine phosphate sodium tetrahydrate" prepared by the present invention is abbreviated as "crystalline hydrate 1" in the following examples "

Example 1

Preparation of crystalline hydrate 1.

Dissolving 20 g of creatine phosphate sodium in 30ml of water, controlling the temperature of the system to be 10 ℃, adding 90ml of methanol under stirring, stirring for 2 hours, carrying out suction filtration, washing the solid with 95% of ethanol, and drying for 2 hours under 50 ℃ of air flow to obtain the crystalline hydrate 1.

Example 2

The procedure of example 1 was followed, with varying experimental parameters, to obtain the product shown in Table 1.

TABLE 1 product conditions obtained under different process conditions

From the preparation results under the different conditions, when the amount of the added methanol serving as the crystallization agent is not more than 3 times of the amount of the water serving as the solvent, the high-crystal-purity creatine phosphate sodium tetrahydrate, namely, crystal hydrate 1, can be obtained; when the amount of the crystallization agent 95% ethanol or absolute ethanol is not more than 3 times of the amount of water used as a solvent, the high-crystal-form-purity creatine phosphate sodium tetrahydrate can be obtained. Under the condition that the water content of the crystallization agent is 3 times more than that of the solvent, the obtained creatine phosphate sodium crystal tetrahydrate with non-high crystal purity is obtained.

Example 3

The crystalline hydrate 1 obtained in example 1, the domestic commercially available sample 2, the domestic commercially available sample 3, and the imported commercially available sample were subjected to an X-ray powder diffraction test and a thermogravimetric analysis (TG) test, respectively.

X-ray powder diffraction experiments: instrument Bruker D8advance, operating condition Cu (40kV, 40mA), scan range 3 ° -40 °, scan step 0.02 °, each step 0.1 seconds.

X-ray powder diffraction experimental results: the pattern of crystalline hydrate 1 shows only the characteristic diffraction peaks of crystalline 4.5 hydrate, see fig. 1. The spectra of the domestic marketed samples 1-3 and the imported marketed samples are mixed with the characteristic diffraction peak of the crystalline 1.5 hydrate in addition to the characteristic diffraction peak of the crystalline 4.5 hydrate, the representative spectrum of the domestic marketed sample is shown in figure 2, and the spectrum of the imported marketed sample is shown in figure 3.

Thermogravimetric (TG) experiments: the temperature of an instrument, NETZSCH TG 209F3, is 30-400 ℃, and the heating rate is 10K/min.

Thermogravimetric (TG) experimental results: the second-stage weight loss of the TG map is shown in Table 2, the second-stage weight loss of the TG map of the crystalline hydrate 1 is less than 5.0%, and the second-stage weight loss of the domestic market samples 1-3 and the imported market samples is more than 5.0%. The TG spectrum of the crystalline hydrate 1 is shown in FIG. 4, the TG spectrum of a representative sample on the market at home is shown in FIG. 5, and the TG spectrum of a sample on the market at import is shown in FIG. 6.

TABLE 2 TG weight loss percentages for different samples

Example 4

The moisture content of the crystalline hydrate 1 obtained in examples 1 and 2 was measured by the karl fischer method, and the measurement results are shown in table 3.

TABLE 3 determination of the Water content of the samples by the Karl Fischer method

Sample (I)	Water content	Sample (I)	Water content
				Example 1 sample	24.22％	Example 2 sample 7	24.16％
Example 2 sample-1	24.68％	Example 2 sample-9	24.23％
				Example 2 sample-2	24.10％	Example 2 sample No. 10	24.35％
Example 2 sample-3	24.85％	Example 2 sample 11	24.66％
				Example 2 sample 6	23.97％

The karl fischer method results show that the moisture content of crystalline hydrate 1, around 24.1%, matches the theoretical moisture content value of creatine phosphate sodium crystalline 4.5 hydrate.

Example 5

Two portions of the crystalline hydrate 1 obtained in example 1 were taken in parallel, one portion was left open in a drying oven at 90 ℃ for 0.5 hour, the other portion was left open in a drying oven at 120 ℃ for 0.5 hour, and then cooled to room temperature in the respective dryers, and X-ray powder diffraction measurement was performed under the conditions of example 3.

As a result: after the crystalline hydrate 1 is dried at 90 ℃ for 0.5 hour, the diffraction peak is almost unchanged, and the diffraction peak of the crystalline hydrate 1.5 does not appear; after drying at 120 ℃ for 0.5 hour, the diffraction peak disappeared and became amorphous as shown in FIG. 7.

Example 6

Taking two parallel samples of 1-3 domestic samples and imported samples, placing one sample in a drying oven at 90 ℃ for 0.5 hour in an open manner, placing the other sample in a drying oven at 120 ℃ for 0.5 hour in an open manner, cooling to room temperature in the drying oven respectively, and performing X-ray powder diffraction measurement according to the conditions of example 3.

As a result: after the domestic and imported samples 1-3 on the market and the imported samples on the market are dried at 90 ℃ for 0.5 hour, the original characteristic diffraction peak of the 4.5 hydrate in the X-ray powder diffraction pattern is weakened to disappear, and the diffraction peak of the crystalline 1.5 hydrate is shown; after drying at 120 ℃ for 0.5 hour, the original characteristic diffraction peak of 4.5 hydrate disappears, and the diffraction peak of 1.5 hydrate appears. A representative map (sample 3 from domestic marketing) is shown in FIG. 8.

Example 7

Sample N (crystalline 1.5 hydrate) was prepared by the method of reference 1. The water content was 10.1% by Karl Fischer method. The X-ray powder diffraction measurement was performed on sample N under the same conditions as in example 3, resulting in: as shown in fig. 9, sample N exhibited a diffraction peak of crystalline 1.5 hydrate.

Example 8

And (5) comparing stability.

The crystalline hydrate 1 obtained in example 1 was packaged in a commercial package, and a sample 1 on the market, a sample 2 on the market, a sample 3 on the market and a sample on the market were imported and placed in a forced air drying oven at 70 ℃ for 2 hours to detect the increase of related substances. The results are shown in Table 4.

TABLE 4 stability data at 70 ℃ for different samples

The results show that: the crystal hydrate 1 has little change in related substances when the crystal hydrate is placed at 70 ℃ for 2 hours,

while the remaining samples increased significantly.

Example 9

And (5) comparing stability.

The crystalline hydrate 1 obtained in example 1 is packaged on the market, and is placed in a constant-temperature drying oven at 60 ℃ together with a domestic market sample-3, and samples are taken on the 0 th day, the 5 th day and the 10 th day for detecting creatine phosphate and related substances. The results are shown in Table 5:

TABLE 5 stability data at 60 ℃ for different samples

The results show that: the crystal hydrate 1 is placed at 60 ℃ for 10 days, the related substances are increased slightly, while the related substances of the samples on the market are increased remarkably, and the main components are decomposed seriously.

Example 10

And (5) performing a crystal form mixing comparison experiment I.

The crystalline hydrate 1 obtained in example 1 and sample N were separately ground through a 200 mesh sieve for use.

Respectively taking a certain amount of screened crystalline hydrate 1 and a certain amount of screened sample N to enable the sample N to account for 2%, 0.5% and 0.05% of the total weight of the two, grinding and uniformly mixing the two by adopting a step-by-step uniformly mixing method, sieving by a 200-mesh sieve, and uniformly mixing. 4 samples were obtained and reported as sample-2%, sample-0.5%, sample-0.05%. The milled crystalline hydrate 1 was added and marked as crystalline hydrate 1-mill. Respectively taking a certain amount of 4 samples, placing the samples in an oven at 120 ℃ for 0.5 hour in an open mode, and cooling the samples to room temperature in a drier to obtain samples-2-120, samples-0.5-120, samples-0.05-120 and crystal hydrates 1-ground-120. A total of 8 samples were subjected to the X-ray powder diffraction test under the same conditions as in example 3. Then, the scanning speed parameters were adjusted to step size 0.02 degree, scanning speed 1.0 sec/step, and the remainder was unchanged, under which condition 1-grinding of crystalline hydrate, sample-0.05% was measured. Thermogravimetric analysis (TG) experiments were performed on the first 4 samples. The results are shown in the following table, and a partial X-ray powder diffraction pattern is shown in FIG. 10.

TABLE 6 XRPD and TG results for different reference amounts

Note: XRPD experiments refer to X-ray powder diffraction experiments

The experimental results showed that the physical properties of the crystalline hydrate 1 were changed even when 0.05% of 1.5 hydrate was mixed. Although no change is observed in the X-ray powder diffraction pattern before heating after mixing, the X-ray powder diffraction pattern after heating at 120 ℃ for 0.5 hour and the weight loss in thermogravimetric analysis (TG) experiment at 150-240 ℃ both lose the original characteristics. This indicates that even the incorporation of a very small amount of the crystalline form of 1.5 hydrate affects the properties associated with the thermal stability of crystalline 4.5 hydrate.

Example 11

And a second crystal form mixing comparison experiment.

Samples from example 10 were taken: and (3) detecting the increase of related substances by adopting a simulated marketed package of sample-2%, sample-0.5%, sample-0.05% and ground and sieved crystal hydrate 1 and sample N, and placing the package in a forced air drying oven at 70 ℃ for 2 hours. The results are as follows:

TABLE 7 stability data for different parameters

The results show that the high crystal purity creatine phosphate tetrahydrate can be significantly reduced in thermal stability even when a very small amount of crystalline 1.5 hydrate is incorporated.

Example 12

1 g of the crystalline hydrate 1 obtained in example 1, an imported and marketed sample, a domestic and marketed sample 1 and a domestic and marketed sample 2 are weighed into weighing bottles, and precisely weighed. And placing the opening in a 50 ℃ forced air drying box, taking out the opening respectively for 1 hour, 4 hours and 7 hours, cooling the opening to room temperature in a drier, precisely weighing the opening, and calculating the weight loss. The results are shown in Table 8.

TABLE 8 loss on drying of different samples

The water content of the crystalline hydrate 1, the imported and marketed samples, the domestic and marketed samples 1 and the domestic and marketed samples 2 measured by a Karl Fischer water content titrator is as follows: 24.22%, 22.43%, 23.16%, 22.67%.

After drying, the weight of the crystal hydrate 1 is basically unchanged when the crystal hydrate is dried at 50 ℃, and the drying process is easy to control. Whereas the marketed sample, when dried at 50 ℃, shows a continuous decrease in weight over 5 hours, with a constant weight after a weight loss of about 14%. However, the water content is not within the range (20.0% to 25.0%) defined by the quality standard, and the drying process is not easy to control.

Example 13

Preparation of crystalline hydrate 1 for injection

Taking 10kg of creatine phosphate sodium 4.5 hydrate, adding 10kg of water for injection, stirring and dissolving, controlling the system temperature at 10 ℃, adding 767 type active carbon to remove a heat source, filtering and sterilizing by using a 0.22 mu m microporous filter membrane, adding 30L of methanol filtered by the 0.22 mu m microporous filter membrane, stirring for 3 hours, adding into a multifunctional filtering, washing and drying machine (three-in-one), filtering, wherein a detergent is 95% ethanol filtered by the 0.22 mu m microporous filter membrane, washing, filtering, drying by introducing 60 ℃ sterilized hot air, and sieving. The sample was left for detection and the water content was 24.3% by Karl Fischer method. 0.66g (containing creatine phosphate sodium 0.5g) per bottle, subpackaging in penicillin bottles, adding rubber plugs and sealing with aluminum caps.

In the retained sample, a certain amount of sample is taken to carry out an X-ray powder diffraction experiment, the experimental conditions are the same as those of example 3, and the result map only shows the characteristic diffraction peak of crystalline 4.5 hydrate and does not have the characteristic diffraction peaks of 1.5 hydrate and 6 hydrate. Another certain amount of the powder was placed in an oven at 120 ℃ for 0.5 hour, cooled to room temperature in a desiccator, and subjected to the X-ray powder diffraction test under the same test conditions as in example 3. The resulting spectrum showed no significant diffraction peaks and was amorphous.

Claims

1. Formula (1) high crystal purity creatine phosphate sodium tetrahydrate

The creatine phosphate sodium tetrahydrate hemihydrate simultaneously meets the following conditions I-III:

2. The high crystalline purity sodium phosphocreatine tetrahydrate of claim 1, wherein: the X-ray powder diffraction pattern is shown in FIG. 1.

3. The high crystalline purity sodium phosphocreatine tetrahydrate of claim 1, wherein: the conventional scanning of the X-ray powder diffractometer described in the condition II means that the scanning is performed at a scanning speed of 1.2 to 12 DEG/min.

4. A method for preparing the high crystalline purity creatine phosphate tetrahydrate of claim 1, comprising the steps of:

the crystallization agent is methanol, and the volume consumption of the crystallization agent is three times or less than three times of the water amount.

5. The method for preparing the high-crystal-purity creatine phosphate tetrahydrate of claim 4, wherein the method comprises the following steps: in the step 1), creatine phosphate sodium is dissolved in water in an amount which is 1-1.5 times that of creatine phosphate sodium, and is stirred and dissolved.

6. The method for preparing the high-crystal-purity creatine phosphate tetrahydrate of claim 4, wherein the method comprises the following steps: and 2) controlling the system temperature at 0-15 ℃, and adding a crystallization agent under stirring.

7. Use of the high crystalline purity creatine phosphate tetrahydrate of claim 1, wherein: the high-crystal-purity creatine phosphate sodium tetrahydrate hemihydrate is applied to preparation of creatine phosphate sodium sterile powder injection.