WO2003018615A1 - Novel crystal of cyclic lipopeptide compound - Google Patents

Abstract

Crystals of the cyclic lipopeptide compound (I) represented by the following chemical structural (I) (I) which can be obtained by dissolving the cyclic lipopeptide compound (I) in a hydrous alcohol or hydrous acetone and then adding ethyl acetate, dichloromethane, acetone, acetonitrile, or a mixture of two or more thereof to the solution with cooling or heating.

Description

 Description New crystal of cyclic lipopeptide compound

 The present invention relates to a novel crystal of a cyclic lipopeptide compound. The cyclic lipopeptide compound (I) of the present invention has the following structural formula:

Indicated by This compound has antimicrobial activity (especially antifungal activity) and 8-1,3-glucan synthase inhibitory activity, and is effective in preventing Pneumocystis carinii infections, such as pneumocystis pneumonia. It is useful for prevention and treatment of various infectious diseases including. Background art

The cyclic lipopeptide compound (I) is a known compound, and is described as a compound of Example 25 in, for example, WO96 / 11210. However, the compound (I) obtained by the method described in Example 25 of the publication was an amorphous compound. Thereafter, as a result of diligent studies, the inventors of the present invention formed compound (I) into a special compound. Succeeded in obtaining the compound of formula (I), and succeeded in obtaining the compound of formula (I) with the B82 type crystal of the compound (I). Called.]

 Disclosure of the invention

Physicochemical properties of B82 type 2 crystal of compound (I)

 The physicochemical properties of the B82 type crystal of the compound (I) obtained by the present invention are described below.

 1) Crystal morphology

 Needle-shaped crystals

 2) X-ray powder diffraction pattern

 The B82 type 2 crystal of the compound (I) shows its characteristic peak in the powder X-ray diffraction pattern at the diffraction angle [2 ° (°)] shown in the following table.

 Diffraction angle (°) Diffraction angle (°)

 Around 4.6 ° Around 19.2 °

 5. Near 4 ° Near 19.6 °

 Around 6.4 ° Around 20.2 °

 Around 7.6 ° Around 20.7 °

 Around 9.0 ° Around 21.3 °

 Around 9.8 ° Around 22.2 °

 Around 11.8 ° Around 22.7 °

 Around 12.6 ° Around 23.3 °

 Around 13.9 ° Around 24.0 °

 Around 15.1 ° around 24.5 °

 Around 15.9 ° Around 25.3 °

 16.around 9 ° around 26.2 °

Around 17.9 ° Around 26.7 ° FIG. 1 shows a powder X-ray diffraction pattern of a B82 type crystal of compound (I) obtained in Example 1 described later.

 However, this diffraction pattern is given only as a reference, and any crystal of compound (I) whose powder X-ray diffraction pattern is essentially the same as this diffraction pattern, It is identified as a B82 type 2 crystal of the compound (I).

 3) Infrared absorption spectrum

 FIG. 2 shows an infrared absorption spectrum of a B82 type crystal of the compound (I) obtained in Example 1 described later.

 However, this spectrum is given for reference only, and any crystal of compound (I) whose infrared absorption spectrum is essentially the same as this spectrum is used. Is also identified as a B82 type 2 crystal of the compound (I).

Method for producing B82 type 2 crystal of compound (I)

 Hereinafter, the method for producing the B82 type crystal of compound (I) obtained by the present invention will be described in detail.

 The B82 type crystal of the compound (I) is obtained by dissolving the compound (I) in aqueous alcohol or aqueous acetone, and cooling or heating the mixture to form ethyl acetate, dichloromethane, or acetone. Alternatively, it can be obtained by post-adding acetonitrile or a mixed solution thereof and precipitating crystals under cooling or heating.

 Preferred examples of the solution containing the compound (I) include a hydrated methanol solution, a hydrated ethanol solution, a hydrated isopropanol solution and a hydrated butanol solution of the compound (I).

The B82 type crystal of the compound (I) is prepared by dissolving the compound (I) in a hydrated alcohol and heating the mixture at room temperature or under heating (preferably at 40 ° C to 40 ° C). At 5 ° C), ethyl acetate, dichloromethane, acetone or acetonitrile or a mixture thereof was added afterwards, stirring was continued slowly, and then the mixture was cooled (preferably 10%). (° C to 5 ° C) to precipitate crystals.

 When the B82 type crystal of the compound (I) is precipitated, it is preferable to keep the solution slightly above the saturated state.

 The B82 type crystal of compound (I) obtained by the above production method is filtered and dried by a conventional method.

 The amorphous compound (I) used in the present invention can be produced by the method described in the above-mentioned WO 96/11210.

 Industrial applicability

 Since the B82 type crystal of the compound (I) is much higher in purity and superior in stability than the amorphous compound (I), the B82 type crystal of the compound (I) is It is easy to handle in formulation and storage.

 As described above, the B82 type crystal of the compound (I) has extremely excellent advantages, and is very superior to the amorphous compound (I).

 BEST MODE FOR CARRYING OUT THE INVENTION In order to show such advantages of the B82 type crystal of the compound (I), the B82 type crystal of the compound (I) and the above-mentioned W096 / 11 The results of comparative experiments on the purity with the amorphous compound (I) described in Japanese Patent Publication No. 210 are described below.

Test sample

 Test sample 1: Amorphous compound (I) obtained in Example 25 in WO966Z11210

Test sample A: B82 type 2 crystal of compound (I) of the present invention Test method

 Impurities were measured by high performance liquid chromatography (HPLC).

 H P L C condition

 Column: Puresil C18 12θΑ

 4.6mm inside diameter, 25cm length (particle size 5 // m)

 Mobile phase: pH 2.5 Perchlorate buffer: Acetonitrile mixed solution (4: 3)

 Force ram temperature: 40. C

 Wavelength: 210nm

 Flow:: 0.5ml / min

 Sample concentration: 0.5mg / mL

 Injection volume: 10μ 1

 Impurity content of each test sample

高脂溶性物質測定 H P L C条件

Measurement of highly fat-soluble substances HPLC conditions

Column: Puresil C18 12θΑ 4.6mm inside diameter, 25cm length (particle size 5 / zm)

Mobile phase: pH 2.5 Perchlorate buffer: Acetonitrile mixed solution (7:13)

Force ram temperature: 40 ° C

Wavelength: 210nm

Flow rate: 1. ral / min

Sample concentration: lmg / mL

Injection volume: 20 1

 Impurity content of each test sample (high fat-soluble substance)

試験結果から明らかなように、 試験サンプル Aは、 試験サンプル 1 に比べて、 不純物含有の割合が少ないことがわかる。

As is clear from the test results, test sample A has a lower impurity content than test sample 1.

 As described above, the purity between the B82 type crystal of the compound (I) and the amorphous compound (I) described in WO966Z11210 A big difference. That is, the compound of the present invention

It has been clarified that the B82 type crystal of (I) is extremely superior to the amorphous compound (I) described in the patent publication. Example

Example 1

 0.5 g of the compound (I) (amorphous form) was dissolved to a concentration of 50 rag / raL using 70% aqueous methanol. Ethyl acetate was added to an aqueous methanol solution in which the compound (I) was dissolved, so that the concentration of the compound (I) was 8 mg / mL (to a concentration of 20 mg / raL after the first addition, and stirred overnight. After aging, a second addition of ethyl acetate was made to a final concentration of 8 rag / mL.) After the addition of the total amount, B82 type 2 crystal of compound (I) was obtained by separation and filtration (0.5 g). This example was performed at normal temperature (25 ° C.). FIG. 1 shows a powder X-ray diffraction pattern of the B82 type 2 crystal. FIG. 2 shows an infrared absorption spectrum (KBr method) of the B82 type crystal.

 The peaks in the powder X-ray diffraction pattern of the B82 type 2 crystal are shown below. The measurement conditions are as follows.

Anti-cathode: Cu, tube voltage: 40 kV, tube current: 30 mA, detector: proportional counter

Diffraction angle ') Diffraction angle (°)

Around 4.6 ° Around 19.2 °

 Around 5.5 ° Around 19.6 °

 Around 6.4 ° Around 20.2 °

 Around 7.6 ° Around 20.7 °

 Around 9.0 ° Around 21.3 °

 Around 9.8 ° Around 22.2 °

 Around 11.8 ° Around 22.7 °

 Around 12.6 ° Around 23.3 °

 Around 13.9 ° Around 24.0 °

 Around 15.1 ° around 24.5 °

 Around 15.9 ° around 25.3 ° '

 Around 16.8 ° Around 26.2 °

 Around 17.9 ° Around 26.7 ° Example 2

 0.2 g of the compound (I) (amorphous) was dissolved at a concentration of 50 mg / mL using 80% aqueous ethanol. Ethyl acetate was added to an aqueous solution of ethanol in which compound (I) was dissolved, so that the concentration of compound (I) was 17 mg / mL. The obtained B82 type 2 crystal was obtained (0.18 g). This example was performed at normal temperature (25 ° C).

Example 3

3.2 g of the compound (I) (amorphous form) was dissolved to a concentration of about 40 mg / mL using 75% aqueous methanol. Methanol was added to an aqueous methanol solution in which the compound (I) was dissolved so that the concentration of the compound (I) was about 85%. Then, cool down to 5 ° C and keep it at 5 ° C. Ethyl acid was added (primary addition) so that the concentration of compound (I) was about 25 mg / mL, and then a separately prepared 1: 1 mixed solution of ethyl acetate / acetone was added to compound (I). Secondary addition was performed over time so that the concentration was about 3 mg / mL. After the addition of the total amount, B82 type crystal of compound (I) was obtained by separation and filtration. (2.2 g)

Example 4

 Compound (I) (amorphous form) was dissolved in 6% rag / mL using 90% acetone water. To the aqueous acetone solution in which this compound (I) was dissolved, 4.4 times the amount of ethyl acetate of this solution was added. After stirring at room temperature (25 ° C) for 3-4 hours, the mixture was heated to 40 ° C, and then cooled while stirring to 5 ° C. After stirring was continued for 18 to 20 hours, B82 type crystal of compound (I) was obtained by separation and filtration.

Example 5

Compound (I) (amorphous) was dissolved at a concentration of 6 mg / mL using 70% aqueous isopropyl alcohol. Ethyl acetate 1.6 times the amount of this solution was added to a solution of the compound (I) in an aqueous solution of isopropyl pill. Room temperature After stirring (25 ° C) at 3-4 hours, warmed to 40 ^D C, then, while stirring until 5 ° C, and then cooled over about 20 hours. Observation of the crystallization solution under a microscope revealed B82 type 2 crystals.

Example 6

 Compound (I) (amorphous form) was dissolved in 90% aqueous methanol to a concentration of 12.5 mg / mL. To a methanol aqueous solution in which this compound (I) was dissolved, 1.5 times the amount of dichloromethane in this solution was added. After stirring at room temperature (25 ° C) for 3-4 hours, the mixture was heated to 40 ° C, and then cooled with stirring to 5 ° C over about 20 hours. When the crystallization solution was observed under a microscope, B82 type 2 crystals were observed.

Example 7 Compound (I) (amorphous) was dissolved at a concentration of 6 mg / mL using 80% aqueous methanol. Acetonitrile, 3.2 times the amount of this solution, was added to an aqueous methanol solution in which this compound (I) was dissolved. After stirring at room temperature (25 ° C) for 3 to 4 hours, the mixture was heated to 40 ° C, and then cooled to about 5 ° C over about 20 hours. Observation of the crystallization solution under a microscope revealed B82 type 2 crystals.

Example 8

 The compound (I) (amorphous form) was dissolved at a concentration of 100 mg / mL using 90% aqueous methanol. An aqueous methanol solution in which this compound (I) is dissolved is stirred at room temperature (25 ° C) for 3-4 hours, then heated to 40 ° C, and then stirred for about 20 hours while stirring at 5 ° C. Cool. Observation of the crystallization solution under a microscope revealed B82 type 2 crystals.

 BRIEF DESCRIPTION OF THE FIGURES

Figure 1 shows the powder X-ray diffraction pattern.

Figure 2 shows the infrared absorption spectrum.

Claims

The scope of the claims 1. A crystal of a cyclic lipopeptide compound (I) represented by the following chemical structural formula, which shows a peak at a diffraction angle shown in the following table in a powder X-ray diffraction pattern:  Diffraction angle (°)  Around 4.6 °  Around 5.4 °  Around 9.0 °  Around 9.8 °  Around 16.9 °

2. The cyclic lipopeptide compound (I) according to claim 1 is dissolved in hydrated alcohol or hydrated acetone, and the mixture is cooled or heated to obtain ethyl acetate, dichloromethane, acetone or acetonitril. Alternatively, crystals of the cyclic lipopeptide compound (I), which can be obtained by post-adding a mixed solution thereof. 3. Dissolve the cyclic lipopeptide compound (I) according to claim 1 in hydrated alcohol or hydrated acetone, and slowly stir at 5 ° (to ~ 45 ° C). Then, after adding thereto ethyl acetate, dichloromethane, acetone or acetonitrile or a mixed solution thereof, 3. The crystal according to claim 2, which can be obtained by stirring and standing at 5 ° C to 45 ° C.  4. The cyclic lipopeptide compound (I) according to claim 1 is dissolved in hydrated alcohol or hydrated acetone, and the mixture is cooled or heated to obtain ethyl acetate, dichloromethane, acetone or acetate. A method for producing crystals of the cyclic lipopeptide compound (I), characterized in that crystals are obtained by post-adding the mixture thereof.  5. Dissolve the cyclic lipopeptide compound (I) according to claim 1 in aqueous alcohol or aqueous acetone, continue stirring slowly at 5 ° C to 45 ° C, and then add After adding ethyl, dichloromethane, acetone or acetonitrile or a mixture thereof, the crystals are obtained by stirring at 5 ° C to 45 ° C and leaving to stand. 4. The production method according to 4.