WO2011118929A2

WO2011118929A2 - Novel crystal forms of epirubicin hydrochloride

Info

Publication number: WO2011118929A2
Application number: PCT/KR2011/001706
Authority: WO
Inventors: Moon-Sung Kim; Soon-Hoe Kim; Jun-Ho Choi; Si-Young Sung; Hong-Gyu Lim; Dae-Won Cha; Byoung-Hwa Son; Ki-Moon Ryu
Original assignee: Dong-A Pharm.Co.,Ltd.
Priority date: 2010-03-24
Filing date: 2011-03-11
Publication date: 2011-09-29
Also published as: KR20110107237A; WO2011118929A3; TW201143777A; KR101125460B1

Abstract

The present invention relates to the crystal form of Epirubicin hydrochloride stable against heat and water and the preparation method thereof. The novel crystal form has the physiochemical property stable against heat and water according to the present invention and it improves the excellent stability of the agents by preventing the relative substances from the one containing it. Furthermore, the crystal form of Epirubicin hydrochloride is suitable for mass production since it is easily prepared.

Description

NOVEL CRYSTAL FORMS OF EPIRUBICIN HYDROCHLORIDE

The present invention relates to a process for preparing 5,12-naphthacenedione, 10-[(3-amino-2,3,6-trideoxy-α-L-arabino-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-8-(2-hydroacetyl)-1-methoxy hydrochloride (hereinafter referred to as Epirubicin hydrochloride) as an anthracycline anticancer agent and its novel crystal forms thereof.

In order to demonstrate the proper effect, drugs are generally requested to prevent the degradation of active ingredient content during the retention (storage) as well as after the production. It is also requested to suppress the increasing impurities as a decomposed material of the active ingredient or the related substances during the same period. Therefore, it is highly important to prevent mixing of impurities in pharmaceutical quality control.

According to the regulation of the national formulary regarding the purity test, the related substance is set aside particularly in the article of the Korean pharmacopoeia. However, unless otherwise specified besides ordinary impurities, the sum of the related substances is decided less than 2.0% or the amount of them is regulated by the related compounds and chromatographic purity in each group in the American pharmacopoeia. In addition, it is disclosed the related substances in the European and the English pharmacopoeia, and its amount is regulated by the related substances on the purity test in the Japanese pharmacopoeia.

The raw material of the new drug is disclosed in the guideline Q3A of ICH which is the international agenda about medicines, and the general information about the set of the impurity standard among the new medical products in Q3B. Furthermore, the impurity standard of products is subdivided into Reporting Thresholds, Identification Thresholds, and Safety Qualification Thresholds.

Therefore, the impurity beyond levels is investigated for evaluating the toxicological effects to establish the basis of deciding the safety for human on administration as well as for its physical and chemical structure. In other words, a great difference in preparing the material is provided in accordance with the rise of the minimum of the impurities, and it also means its great increase of the potentially toxicological effects.

Thus, the control of impurities contained in drugs is addressed in the important matter in the international agreements as well as the individual countries.

Epirubicin hydrochloride as active ingredient of the present invention is known to be useful in the treatment of breast cancer, and it is also the drug lowering the hepatotoxicity which anthracyclin antitumor agents contain. The drug mechanism and action has already been disclosed in detail, and its structure is as shown in the following Formula 1.

However, it is required to be careful in handling Epirubicin hydrochloride since it is unstable against heat and water, and the high-stabled agent is required when formulation. Due to the instability of Epirubicin hydrochloride, impurities are easily generated.

U.S. patent publication No.4,112,076, No.4,345,068, and No.4,861,870 and No.5,945,518 disclose the process for preparing Epirubicin hydrochloride.

U.S. patent publication No.4,112,076 discloses the process for preparing Epirubicin hydrochloride in solid state obtained by the difference of solubility caused by ionic bonding between Epirubicin base dissolved in organic solvent and hydrochloride. U.S. patent publication No.4,861,870 describes the method for preparing Epirubicin hydrochloride in solid state, wherein acetone is added to Epirubicin hydrochloride dissolved in an aqueous solution or a mixed solution of the aqueous solution with organic solvents, however, Epirubicin hydrochloride manufactured in the prior art is amorphous.

Amorphous Epirubicin hydrochloride is not suitable for mass production in filtration and drying, and it should be protected from heat, light, oxygen, and moisture due to its physical and chemical property. Thus, in case of Epirubicin hydrochloride preparation, it is significant to control humidity of the workspace when exposed to the packaging due to its property to condense moisture, as well as it requires to be careful of setting the temperature during the process and drying due to its low heat stability.

U.S. patent publication No.7,485,707 discloses 'Epirubicin hydrochloride crystal form Ⅱ', wherein it is known for the good heat stability thereof.

Meanwhile, the above Epirubicin hydrochloride crystal form Ⅱ doesn`t ensure the stability against water.

Epirubicin hydrochloride crystal form Ⅱ is prepared by adding much insoluble solvent, 1-propanol, after dissolving amorphous Epirubicin hydrochloride in the small amount of water and ethanol. However, the solvent of 1-propanol is slightly soluble and dissolve Epirubicin hydrochloride crystal form Ⅱ as the end product of crystallization and it causes the low yield of Epirubicin hydrochloride crystal form Ⅱ, accordingly, it is not suitable for commercial use.

Moreover, U.S. patent publication No.7,485,707 discloses the method for preparing Epirubicin hydrochloride crystal form Ⅱ does not stay in a gelled phase and is formed into solid within a few seconds or minutes in the period of concentration after dissolving ethanol in water.

Accordingly, it is needed for the person skilled in the art to provide the novel crystal form which is physiochemically stable to the various forms of Epicubicin hydrochloride and easy to prepare.

Thus, it is great significant to maintain the rate of impurities below the certain level during the storage of agents containing Epirubicin hydrochloride. It has advantages to reduce the risk of toxic from impurities as well as to bring economic benefits by preventing additional physiochemical structure of the related substances and/or toxicological study.

The present inventors have made attempts to improve the stability of the agent of Epirubicin hydrochloride by inhibiting the related substance from the agent having heat and water stability, and develop the method for preparing the crystal form of Epirubicin hydrochloride which is easily prepared.

Accordingly, the present invention is designed to provide the crystal form of Epirubicin hydrochloride having the stability against heat and water physiochemically, and to improve the stability of Epirubicin hydrochloride agent by preventing generating the related substance.

Another object of the present invention is to provide the process for preparing the novel crystal form easily and obtaining it in high yield and high purity.

In order to solve the problems, the present invention provides the novel crystal form A, B, C, and D of Epirubicin hydrochloride and their manufacturing methods.

The present invention is designed to provide the crystal form A of Epirubicin hydrochloride and its manufacturing method.

In the present invention, Epirubicin hydrochloride has 2θ of the diffraction angle as shown in the following table 1, and the value in excess 10% of the relative intensity.

As shown in figure 1, in the weak intensity such as 5.15, 17.61, 20.51, 24.42, 25.12, 30.78, 31.95, 33.07, 38.75, 41.06, 42.03, and 42.58°of 2θ, the crystal form A of Epirubicin hydrochloride has additional X-ray powder diffraction peaks, beside them listed in table 1. Herein, "weak intensity" means the peaks less than 10 % of relative intensity as shown in table 1 below. The reference, X-ray diffraction procedures by H.P. Klug and L.E. Alexander, and J. Wiley, New Work(1974) disclose the theory regarding the X-ray powder diffraction pattern.

The crystal form A of Epirubicin hydrochloride of the present invention has a melting point of 183.40℃ as the result of differential scanning calorimetry through DSC.

In the present invention, the crystal form A of Epricubicin hydrochloride is prepared by crystallization in room temperature lowered to slowly after dissolving the amorphous form of Epirubicin hydrochloride in N,N-dimethyl form amide and suspending in alcohol and dissolving in 60~100℃.

The process for preparing the crystal form A of Epirubicin hydrochloride is carried out using alcohol and it is preferred to employ the low-grade alcohol having 1 to 6 carbon atoms. It is more preferred to use 1-butanol. Furthermore, it is possible to mix N,N-dimethyl form amide with alcohol in the volume ratio of 1:15 to 1:50.

Table 1

Moreover, the present invention provides the crystal form B of Epirubicin hydrochloride and the manufacturing process thereof.

In the present invention, it is seen that the crystal form B of Epirubicin hydrochloride has 2θ of diffraction angle and 10% or more of relative intensity as listed in table 2.

Besides the X-ray powder diffraction peaks listed in table 2, it is revealed that the crystal form B of Epirubicin hydrochloride has additional X-ray powder diffraction peaks in as the weak intensity such as 37.55, 38.58, 40.38˚of 2θ as shown in figure 2.

The crystal form B of Epirubicin hydrochloride of the present invention has a melting point of 183.43℃ as the result of differential scanning calorimetry through DSC.

Table 2

In the present invention, the crystal form B of Epricubicin hydrochloride is prepared by crystallization in room temperature lowered to slowly after suspending in alcohol and dissolving in 60~100℃, adding a small amount of water.

The process for preparing the crystal form B of Epirubicin hydrochloride is carried out using alcohol and it is preferred to employ the low-grade alcohol having 1 to 6 carbon atoms. It is more preferred to use 1-butanol.

Moreover, the present invention provides the crystal form C of Epirubicin hydrochloride and the manufacturing process thereof.

In the present invention, it is seen that the crystal form C of Epirubicin hydrochloride has 2θ of diffraction angle and 10% or more of relative intensity as listed in table 3.

Besides the X-ray powder diffraction peaks listed in table 3, it is revealed that the crystal form C of Epirubicin hydrochloride has additional X-ray powder diffraction peaks in as the weak intensity such as 39.41, 45.61, 50.40˚of 2θ as shown in figure 3.

The crystal form C of Epirubicin hydrochloride of the present invention has a melting point of 190.42℃ as the result of differential scanning calorimetry through DSC.

In the present invention, the crystal form C of Epirubicin hydrochloride is manufactured by crystallization of the compound of the crystal form A or B, suspended in a solvent selected from the group consisting of acetone, isopropyl ether, and acetonitril

Table 3

Furthermore, the present invention provides the crystal form D of Epirubicin hydrochloride and the manufacturing process thereof.

In the present invention, it is seen that the crystal form D of Epirubicin hydrochloride has 2θ of diffraction angle and 10% or more of relative intensity as listed in table 4.

It is revealed that the crystal form D of Epirubicin hydrochloride has additional X-ray powder diffraction peaks in as the weak intensity such as 10.21, 31.05, 36.34, 38.24, and 40.19˚ of 2θ as shown in figure 3 including the x-ray powder diffraction peaks listed in table 4.

The crystal form D of Epirubicin hydrochloride of the present invention has a melting point of 192.98℃ as the result of differential scanning calorimetry through DSC.

Table 4

The crystal form D of Epirubicin hydrochloride is prepared by crystallization after dissolved in the small amount of water to melt the amorphous form of Epirubicin hydrochloride and suspended in a solvent selected from the group consisting of acetone, ethyl acetate, acetonitril, and methyl ethyl ketone. Lastly, it is crystallized after dissolved in the temperature of 60~100℃ and cooled down to room temperature slowly.

Accordingly, the novel crystal form A, B, C, and D of Epirubicin hydrochloride are different from the amorphous form and Epirubicin hydrochloride crystal form Ⅱ disclosed in U.S. patent publication No. 7,485,707 on investigating with the X-ray powder diffraction of table 1~4, differential scanning calorimetry, and confirming by microscope.

The value of X -ray diffraction of the present invention is measured by using X`Pert PRO X-ray diffraction system of Panalytical, differential scanning calorimetry is done by using DSC-2010 spectrometer of TA instrument, and the microscope is VHX-600E of Keynence.

The crystal forms of this present invention are disclosed in U.S. patent publication No.4,112,076, No.4,345,068, No.4,861,870, and No.5,945,518, however, it is very advantageous in production, wherein it has better physiochemical stability against heat and moisture than currently marketed amorphous form.

It means the amorphous form of Epirubicin hydrochloride of the prior art is significant to control humidity of the workspace when exposed to the packaging due to its property to condense moisture, as well as it requires to be careful of setting the temperature during the process and drying due to its low heat stability.

However, the crystal form A, B, C, and D of the present invention are suitable to shorten the working hours and being convenient to package or dry under the large range of temperature including room temperature due to its stability against heat. Furthermore, it is unnecessary to be careful to moisture in workspace due to its stability against it. For instance, the crystal form A, B, C, and D could be produced at 60% of the relative humidity in indoor temperature while it is required to control the relative humidity to less than 20% during packaging and drying after the crystallization. Therefore, the process of the crystal form A, B, C, and D of the present invention is more convenient and safe.

The crystal form A, B, C, and D of Epirubicin hydrochloride are the compounds which is possible to use pharmaceutically and already used as oral agent as disclosed in other references. In case of non-oral agents, it is administrated to vein, artery, and bladder, including the injection packed in vials by lypophilization after dissolved in sterile aqueous solution and possibly mixed with diluting agent if necessary.

The crystal forms of Epirubicin hydrochloride is possibly used to produce the anticancer agent by inhibiting the related substances since it has the stability against heat and water. Moreover, it is easily manufactured in high yield and high purity as well as it is suitable for mass production.

Figure 1 is a diagram illustrating X-ray powder diffraction of the crystal form A of Epirubicin hydrochloride.

Figure 2 is a diagram illustrating X-ray powder diffraction of the crystal form B of Epirubicin hydrochloride.

Figure 3 is a diagram illustrating X-ray powder diffraction of the crystal form C of Epirubicin hydrochloride.

Figure 4 is a diagram illustrating X-ray powder diffraction of the crystal form D of Epirubicin hydrochloride.

Figure 5 is a spectral graph illustrating differential scanning calorimetry of the crystal form A of Epirubicin hydrochloride.

Figure 6 is a spectral graph illustrating differential scanning calorimetry of the crystal form B of Epirubicin hydrochloride.

Figure 7 is a spectral graph illustrating differential scanning calorimetry of the crystal form C of Epirubicin hydrochloride.

Figure 8 is a spectral graph illustrating differential scanning calorimetry of the crystal form D of Epirubicin hydrochloride.

Figure 9 is a microphotograph showing the crystal form A of Epirubicin hydrochloride.

Figure 10 is a microphotograph showing the crystal form B of Epirubicin hydrochloride.

Figure 11 is a microphotograph showing the crystal form C of Epirubicin hydrochloride.

Figure 12 is a microphotograph showing the crystal form D of Epirubicin hydrochloride.

This invention will now be described by reference to the following examples which merely illustrative and which are not construed as a limitation of the scope of this invention.

Example 1: Preparation of the crystal form A of Epirubicin hydrochloride

1 g of the amorphous compound of Epirubicin hydrochloride obtained by the method of U.S. patent publication No.4,861,870 dissolve in 20 ml of N,N-dimethyl form amide and disperse and suspend in 20 ml of 1-butanol and higher the temperature until it dissolve completely.(temperature range is approximately 60~100℃) Lastly, it is cooled to room temperature and filtered after suspending. The crystal form of Epirubicin hydrochloride is obtained by vacuum dehydrating the filtered crystal form at 50℃ for 12 hours. (80% of yield)

The crystal form A of Epirubicin hydrochloride is designated, wherein it is measured by X-ray powder diffraction value (using Cuke radiation on M18HF-SRA diffractometer (by Mac Science)) as shown in table 1 and figure 1.

Melting point (measured by DSC-2010 spectrometer of TA instrument; differential scanning calorimetry) : 183.40℃

Example 2: Preparation of the crystal form B of Epirubicin hydrochloride

1 g of the amorphous compound of Epirubicin hydrochloride obtained by the method of U.S. patent publication No.4,861,870 suspend in 200 ml of 1-butanol and add 2 ml of water, stirring it and dissolve until melted completely by increasing temperature(approximately 60~100℃). Lastly, it is cooled to room temperature and filtered. The crystal form of Epirubicin hydrochloride is obtained by vacuum dehydrating the filtered crystal form at 50℃ for 12 hours. (91% of yield)

The crystal form B of Epirubicin hydrochloride is designated, wherein it is measured by X-ray powder diffraction value (using Cuke radiation on M18HF-SRA diffractometer (by Mac Science)) as shown in table 2 and figure 2.

Melting point (measured by DSC-2010 spectrometer of TA instrument; differential scanning calorimetry) : 183.43℃

Example 3: Preparation of the crystal form C of Epirubicin hydrochloride

1 g of the crystal form A of Epirubicin hydrochloride obtained by Example 1 suspend in 50 ml of acetone and filter after stirring it at room temperature. The crystal form of Epirubicin hydrochloride is obtained by vacuum dehydrating the filtered crystal form at 25℃ for 12 hours. (95% of yield)

The crystal form C of Epirubicin hydrochloride is designated, wherein it is measured by X-ray powder diffraction value (using Cuke radiation on M18HF-SRA diffractometer (by Mac Science)) as shown in table 3 and figure 3.

Melting point (measured by DSC-2010 spectrometer of TA instrument; differential scanning calorimetry) : 190.42℃

Example 4: Preparation of the crystal form C of Epirubicin hydrochloride

The crystal form B of Epirubicin hydrochloride obtained from the example 2 is prepared by using the same method as the example 3.(93% of yield) The above-identified crystal form is the same with the crystal form C obtained by the example 3, confirming by measuring with Cuke radiation on M18HF-SRA diffractometer (by Mac Science).

Example 5: Preparation of the crystal form D of Epirubicin hydrochloride

1 g of the amorphous compound of Epirubicin hydrochloride obtained by the method of U.S. patent publication No.4,861,870 dissolve in 10 ml of water and suspend in 400 ml of acetone, stirring it and dissolve until melted completely by increasing temperature(approximately 60~100℃). Lastly, it is cooled to room temperature and filtered. The crystal form of Epirubicin hydrochloride is obtained by vacuum dehydrating the filtered crystal form at 40℃ for 6 hours. (85% of yield)

The crystal form D of Epirubicin hydrochloride is designated, wherein it is measured by X-ray powder diffraction value (using Cuke radiation on M18HF-SRA diffractometer (by Mac Science)) as shown in table 4 and figure 4.

Melting point (measured by DSC-2010 spectrometer of TA instrument; differential scanning calorimetry) : 192.98℃

According to Examples 1 to 5, the present invention provides the process for preparing the novel crystal form of Epirubicin hydrochloride. Using the said method, the crystal form is easily manufactured with high purity and 80~95% of high yield.

Experimental example 1: Evaluation of stability

100 mg of the crystal form A, B, C, and D of Epirubicin hydrochloride prepared in Examples 1, 2, 3, and 5 and 100 mg of the amorphous form of Epirubicin hydrochloride prepared by using the method of U.S. patent publication No.4,861,870 were put into the glass vials respectively. After the vials were sealed with rubber stopper and aluminum cap, the acceleration experiment was carried out, putting them in a thermo-hygrostat (25±2℃, 60% of relative humidity).

In the condition of the above acceleration test, the changes of the amount and the relative substances were evaluated by using HPLC system for 2, 4, and 6 months. The results are listed in the following Table 5, 6, 7, 8, and 9.

[Corrected under Rule 26 21.03.2011]
Table 5

* Unit : %

* Content : use an anhydride removed residual solvents to calculate

* Relative substance A : Doxorubicinone

[Corrected under Rule 26 21.03.2011]
Table 6

* Unit : %

* Content : use an anhydride removed residual solvents to calculate

* Relative substance A : Doxorubicinone

[Corrected under Rule 26 21.03.2011]
Table 7

* Unit : %

* Content : use an anhydride removed residual solvents to calculate

* Relative substance A : Doxorubicinone

[Corrected under Rule 26 21.03.2011]
Table 8

* Unit : %

* Content : use an anhydride removed residual solvents to calculate

* Relative substance A : Doxorubicinone

[Corrected under Rule 26 21.03.2011]
Table 9

* Unit : %

* Content : use an anhydride removed residual solvents to calculate

* Relative substance A : Doxorubicinone

- Column : Zorbax TMS 4.6 mm X 250 mm, 5μm, or its equivalent column

- Mobile phase : mix in a ratio of 17 methanol : 29 acetonitril : 54 buffer solution

- Buffer solution : 3.7 g/L of an aqueous sodium laurilsulfate solution including 2.8% of diluted phosphoric acid

- Detector : UV absorption spectrophotometer (wavelength : 254 nm)

- Flow rate : 2.5 mL/min

- Column temperature : 35℃ ± 2℃

As a result, the contents of the crystal forms of Epirubicin hydrochloride of the present invention are maintained in the acceleration experiment under ±25℃ and 60% of humidity. Also, it has the adequate stability since the contents standards are within 98~100.2% both under the accelerated and harsh conditions. Furthermore, it is revealed that the relative substance A (doxorubicinone) generated from thermal decomposition has thermal due to its lower increase. Therefore, the crystal forms of the present invention show the excellent stability.

Thus, the present invention could improve the stability of Epirubicin hydrochloride agent by providing the crystal form which is stable against heat and moisture, and by preventing the relative substance from the agent containing Epirubicin hydrochloride. Also, it could maintain the rate of impurities less than a certain level during the storage period of the agents containing Epirubicin hydrochloride. It is significant to remove toxicological risk caused by the impurities.

Claims

The crystal form A of Epirubicin hydrochloride having the X-ray powder diffraction peaks as follows:
The crystal form A of Epirubicin hydrochloride according to claim 1, wherein it has additional X-ray powder diffraction peaks in the weak intensity such as 5.15, 17.61, 20.51, 24.42, 25.12, 30.78, 31.95, 33.07, 38.75, 41.06, 42.03, and 42.58°of 2θ
The crystal form B of Epirubicin hydrochloride having the X-ray powder diffraction peaks as follows:
The crystal form B of Epirubicin hydrochloride according to claim 3, wherein it has additional X-ray powder diffraction peaks in the weak intensity such as 37.55, 38.58, 40.38˚of 2θ
The crystal form C of Epirubicin hydrochloride having the X-ray powder diffraction peaks as follows:
The crystal form C of Epirubicin hydrochloride according to claim 5, wherein it has additional X-ray powder diffraction peaks in the weak intensity such as 39.41, 45.61, and 50.40˚of 2θ
The crystal form D of Epirubicin hydrochloride having the X-ray powder diffraction peaks as follows:
The crystal form D of Epirubicin hydrochloride according to claim 7, wherein it has additional X-ray powder diffraction peaks in the weak intensity such as 10.21, 31.05, 31.80, 36.34, 38.24, and 40.19˚of 2θ
The process for preparing the crystal form A of Epirubicin hydrochloride comprising;

(a) dissolving the amorphous form of Epirubicin hydrochloride in N,N-dimethyl form amide;

(b) suspending it in the low-grade alcohol having 1 to 6 carbon atoms;

(c) stirring and dissolving the above suspension at a temperature from 60℃ to 100℃; and

(d) cooling it in room temperature.
The process for preparing the crystal form A of Epirubicin hydrochloride according to claim 9, wherein the low-grade alcohol is 1-butanol.
The process for preparing the crystal form B of Epirubicin hydrochloride comprising;

(a) dissolving the amorphous form of Epirubicin hydrochloride in solvents of the low-grade alcohol having 1 to 6 carbon atoms;

(b) adding water to the suspension;

(c) stirring and dissolving it at a temperature from 60℃ to 100℃; and

(d) cooling it in room temperature.
The process for preparing the crystal form B of Epirubicin hydrochloride according to claim 11, wherein the low-grade alcohol is 1-butanol.
The process for preparing the crystal form C of Epirubicin hydrochloride comprising;

(a) suspending the crystal form A or B of Epirubicin hydrochloride in the solvents selected from the group consisting of acetone, isopropyl ether, and acetonitril; and

(b) stirring and cooling the suspension in room temperature.
The process for preparing the crystal form C of Epirubicin hydrochloride according to claim 13, wherein the solvent is acetone.
The process for preparing the crystal form D of Epirubicin hydrochloride comprising;

(a) dissolving the amorphous form of Epirubicin hydrochloride in the minimum of water;

(b) suspending it in the solvents selected from the group of consisting of acetone, ethyl acetate, acetonitril, and methyl ethyl ketone; and

(c) stirring and dissolving the suspension at a temperature from 60℃ to 100℃ and cooling it in room temperature.
The process for preparing the crystal form D of Epirubicin hydrochloride according to claim 15, wherein the solvent is acetone.
The anticancer composition containing pharmaceutically approved diluting agent, wherein the active ingredient is selected from any crystal form of claim 1 through 8.