An adipate crystal and a preparation method thereof

Abstract

The present invention relates to an adipate crystal of a PI3Kδ / γ dual inhibitor compound (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one compound and a preparation method thereof. The crystal of the present invention is more excellent in physical properties and pharmacodynamic / pharmacokinetic properties.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceuticals, and particularly to an adipate crystal of a PI3Kδ / γ dual inhibitor compound (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one compound and a preparation method thereof. Background Art

[0002] Phosphoinositide-3-kinase (PI3K) belongs to a class of intracellular lipid kinases that phosphorylate the 3-position hydroxyl group of the inositol ring of phosphoinositide lipids (PI), thereby generating lipid second messengers. It has been reported in the prior art that targeted inhibitors of the phosphoinositide-3-kinase (PI3K) pathway can act as immunomodulators.

[0003] A major problem in the large-scale production of pharmaceutical compounds is that the active substance should have stable crystalline polymorphs to ensure consistent processing parameters and drug quality. If an unstable crystalline form is used, the crystal polymorphs may change during manufacturing and / or storage, leading to quality control problems and formulation irregularities. Such changes may affect the reproducibility of the manufacturing method, resulting in the final formulation not meeting the high quality and strict requirements imposed on the formulation of pharmaceutical compositions. In this regard, it should generally be noted that any change in the solid state of a pharmaceutical composition that improves its physical and chemical stability confers a significant advantage over a less stable form of the same drug. In addition, it is crucial to develop a stable production method that always produces the active substance. The existence of multiple crystalline forms with similar solubilities poses a difficult challenge in the large-scale manufacture of pharmaceutical compounds.

[0004] When a compound crystallizes from a solution or slurry, it can crystallize in different spatial lattice arrangements, a property known as "polymorphism". Each crystal form is called a "polymorph". Although the polymorphs of a given substance have the same chemical composition, they can differ from each other in one or more physical properties such as solubility, dissociation degree, true density, dissolution, melting point, crystal shape, compaction behavior, flow properties, and / or solid-state stability.

[0005] Generally as described above, the polymorphic behavior of drugs can be very important in pharmacology. Differences in the physical properties exhibited by polymorphs affect practical parameters such as storage stability, compressibility, and density (important in formulation and product manufacturing), as well as dissolution rate (an important factor determining bioavailability). Changes in chemical reactivity (e.g., differential oxidation, causing discoloration to occur more rapidly when the dosage form is one polymorph than when it is another), mechanical changes (e.g., tablets crumbling after storage as the kinetically favored polymorph converts to the thermodynamically more stable polymorph), or both (e.g., tablets of one polymorph being more prone to decomposition at high humidity) can result in differences in stability. Additionally, the physical properties of crystals can be important in processing. For example, one polymorph may be more likely to form solvates, causing the solid form to aggregate and increasing the difficulty of solid handling. Or, the particle shape and size distribution of one polymorph may differ from another, leading to increased challenges in filtering the drug substance to remove impurities.

[0006] Although drug formulations with improved chemical and physical properties are desired, there are no predictable means to prepare new drug forms (e.g., polymorphs and other new crystalline forms) of existing molecules for such formulations. These new forms would provide consistency in physical properties across a range of environments common in manufacturing and composition use.

[0007] The patent document WO2014195888A1 discloses the (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one compound, whose structural formula is Formula I, and shows a dual PI3Kδ / γ inhibitory function as a free base.

[0008]

[0009] However, there is still an urgent desire to develop a dual PI3Kδ / γ inhibitor that has more excellent properties in terms of physical properties and pharmacodynamics / pharmacokinetics compared to (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one and has high suitability as a pharmaceutical. Summary of the Invention

[0010] The present invention provides a crystalline form of the adipate salt of the compound (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one, characterized in that the crystalline form shows peaks at diffraction angles 2θ of 5.09°, 12.26°, 10.31°, 20.41°, 27.02°, 6.90°, 21.67°, 14.62°, 8.98°, 18.92°, 13.63°, 3.46°, 7.26°, 15.15°, 17.59°, 10.67° (±0.2°) in the X-ray diffraction pattern.

[0011] Furthermore, the crystalline form further includes peaks at diffraction angles 2θ of 4.77°, 22.83°, 28.99°, 23.13°, 19.96°, 16.98°, 14.06°, 27.84°, 18.18°, 16.31°, 25.48°, 24.68°, 21.30°, 26.51°, 30.38°, 19.35°, 11.75° (±0.2°).

[0012] Furthermore, the crystalline form shows an X-ray diffraction pattern substantially as Figure 5 shown.

[0013] In addition, the present invention also provides a method for preparing a crystalline form of the adipate salt of the compound (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one, characterized by comprising the following steps:

[0014] Step (1): Weigh a certain mass of the free base compound of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one and add it to a certain volume of ethyl acetate to obtain suspension 1;

[0015] Step (2): Weigh a certain mass of the adipic acid ligand, dissolve it in a certain volume of ethanol by ultrasonic heating to obtain solution 1;

[0016] Step (3): Dropwise add the solution 1 obtained in step (2) to the suspension 1 obtained in step (1) under stirring until it becomes clear, stir overnight at a low temperature, and no solid is precipitated to obtain solution 2;

[0017] Step (4): Add an anti-solvent to the solution 2 obtained in step (3) to precipitate a solid, and stir at a low temperature to obtain suspension 2;

[0018] Step (5): Centrifuge the suspension 2 obtained in step 4, and vacuum-dry the obtained solid at room temperature to obtain the crystalline form.

[0019] In a preferred technical solution of the present invention, in step (1), the molar ratio of the free base compound to the adipic acid ligand in step (2) is 1:1 - 1.5.

[0020] In a preferred technical solution of the present invention, in step (1), the molar volume ratio of the free base compound to ethyl acetate is: 1 - 10 mol∶0.1 - 1 L.

[0021] In a preferred technical solution of the present invention, in step (2), the molar volume ratio of the adipic acid ligand to ethanol is 1 - 10 mol∶0.1 - 1 L. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Attached Figure 1 XRPD diagram of the free base compound;

[0023] Attached Figure 2 TGA diagram of the free base compound;

[0024] Attached Figure 3 DSC diagram of the free base compound;

[0025] Attached Figure 4 of the free base compound 1 HNMR diagram;

[0026] Attached Figure 5 XRPD diagram of the adipate Form 1;

[0027] Attached Figure 6 TGA diagram of the adipate Form 1;

[0028] Attached Figure 7 DSC diagram of the adipate Form 1;

[0029] Attached Figure 8 DVS diagram and isothermal adsorption curve of the adipate Form 1;

[0030] Attached Figure 9 PLM diagram of the adipate Form 1;

[0031] Attached Figure 10 of the adipate Form 1 1 HNMR diagram;

[0032] Attached Figure 11 XRPD diagram of the hemisuccinate;

[0033] Attached Figure 12 of the hemisuccinate 1 HNMR diagram;

[0034] Attached Figure 13XRPD pattern of phosphate;

[0035] Attached Figure 14 XRPD pattern of oxalate;

[0036] Attached Figure 15 XRPD pattern of hydrobromide; Detailed implementation manners

[0037] I. Analysis methods

[0038] 1.1 X-ray powder diffractometer (XRPD)

[0039] The crystal form of the sample was analyzed using an X-ray powder diffractometer. The 2θ scanning angle of the sample was from 3° to 40°, the scanning step was 0.02°, and the scanning time for each step was 0.2 s. The tube voltage and current were 40 kV and 40 mA respectively. When preparing the sample, an appropriate amount of the sample was placed on the sample stage to ensure that the surface of the sample was smooth and flat.

[0040] 1.2 Differential scanning calorimetry (DSC)

[0041] The sample was analyzed using a TA instruments Q200 DSC. The weighed sample (0.5 mg - 5 mg) was placed in the sample stage, and the sample was heated to the final temperature at a rate of 10 °C / min under the protection of nitrogen (50 mL / min).

[0042] 1.3 Thermogravimetric analysis (TGA)

[0043] The sample was analyzed using a TA instruments Q500. The sample was placed in a platinum crucible with the tare weight removed, and the system automatically weighed it. Then, the sample was heated to the final temperature at a rate of 10 °C / min under the protection of nitrogen (40 mL / min).

[0044] 1.4 Polarizing light microscope (PLM)

[0045] The sample was analyzed using a polarizing light microscope, and the morphology and microstructure of the crystal were obtained by adjusting different magnifications.

[0046] 1.5 Dynamic vapor sorption (DVS)

[0047] Dynamic vapor sorption was performed using a TA Instruments Q5000 SA. Approximately 1 - 10 mg of the sample was placed on the sample pan and suspended in the sample chamber. The temperature in the chamber was maintained at a constant 25 ± 1 °C by a water bath. In step mode, the sample was cycled at a relative environmental humidity of 0% RH - 80% RH. The analysis was performed at 10% RH / step. The time for maintaining each humidity was set to 90 min to allow the sample to equilibrate with the chamber environment.

[0048] 1.6 Liquid nuclear magnetic resonance hydrogen spectrum ( 1 H NMR)

[0049] The sample was analyzed using a Bruker Ascend 500 MHz, with deuterated dimethyl sulfoxide as the solvent.

[0050] II. Preparation method

[0051] 2.1 Preparation method of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one free base compound

[0052] The (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one free base compound was prepared by referring to the method in Example of Patent Document CN105358560A. Among them, the XRPD pattern of the free base compound is as shown in the appendix Figure 1 shown, the TGA pattern is as shown in the appendix Figure 2 shown, the DSC pattern is as shown in the appendix Figure 3 shown, 1 The HNMR pattern is as shown in the appendix Figure 4 shown.

[0053] 2.2 Preparation of the hexanedioate crystalline form of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one

[0054] Step 1: Add about 100 mg of the free base compound prepared in Example 2.1 to 2.0 mL of ethyl acetate to obtain suspension 1;

[0055] Step 2: About 40 mg of adipic acid was dissolved in 0.4 mL of ethanol by ultrasonic heating to obtain solution 1;

[0056] Step 3: Solution 1 was added dropwise to the stirring suspension 1, and it became clear. Stir overnight at 4 °C, and no solid was precipitated to obtain solution 2;

[0057] Step 4: Add about 20 mL of n-heptane to solution 2, and a solid was precipitated. Stir at 4 °C for about 8 h to obtain a suspension;

[0058] Step 5: The suspension was centrifuged, and the obtained solid was vacuum dried at room temperature overnight to obtain the hexanedioate.

[0059] The product was characterized, and its XRPD pattern is as shown in the appendix Figure 5 shown, the TGA and DSC patterns are respectively as shown in the appendix Figure 6 and the appendix Figure 7 shown, the DVS is as shown in the appendix Figure 8 shown, the PLM is as shown in the appendixFigure 9 as shown 1 The \(^1\)H NMR is as attached Figure 10 as shown

[0060] Preparation of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one hemisuccinate

[0061] Step 1: Add about 50 mg of the free base compound prepared in Example 2.1 to 0.5 mL of isopropanol, heat in a water bath at 60 °C until dissolved to obtain Solution 1;

[0062] Step 2: Add about 16 mg of succinic acid to 0.2 mL of isopropanol to obtain Suspension 1;

[0063] Step 3: At room temperature, slowly add Suspension 1 dropwise to Solution 1 until dissolved, stir overnight at 4 °C, no solid precipitates, to obtain Solution 2;

[0064] Step 4: Add about 5 mL of n-heptane to Solution 2, a solid precipitates, stir overnight at room temperature to obtain Suspension 2;

[0065] Step 5: Centrifuge Suspension 2, and vacuum dry the obtained solid overnight at room temperature to obtain the hemisuccinate salt.

[0066] Characterize the product, its XRPD pattern is as attached Figure 11 as shown 1 The \(^1\)H NMR is as attached Figure 12 as shown

[0067] Preparation of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one phosphate

[0068] Step 1: Add about 50 mg of the free base compound prepared in Example 2.1 to 0.5 mL of isopropanol, heat in a water bath at 60 °C until dissolved to obtain Solution 1;

[0069] Step 2: Slowly add about 16 mg of phosphoric acid (dissolved in 132 μL of isopropanol) dropwise to Solution 1, a solid precipitates, stir overnight at 4 °C to obtain a suspension;

[0070] Step 3: Centrifuge the suspension, and vacuum dry the obtained solid overnight at room temperature to obtain the phosphate salt.

[0071] Characterize the product, its XRPD pattern is as attached Figure 13 as shown

[0072] Preparation of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one oxalate

[0073] Step 1: Add approximately 50 mg of the free base compound prepared in Example 2.1 to 0.5 mL of isopropanol, and heat in a water bath at 60 °C until dissolved to obtain Solution 1.

[0074] Step 2: Add approximately 17 mg of oxalic acid to 0.2 mL of ethanol to obtain Solution 2.

[0075] Step 3: At room temperature, slowly add Solution 2 dropwise to Solution 1, stir overnight at 4 °C to precipitate a solid, and obtain a suspension.

[0076] Step 4: Centrifuge the suspension, and vacuum dry the obtained solid overnight at room temperature to obtain the oxalate salt.

[0077] The product was characterized, and its XRPD pattern is shown in the appendix Figure 14 as follows.

[0078] 2.6 Preparation of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one hydrobromide

[0079] Step 1: Add approximately 50 mg of the free base compound prepared in Example 2.1 to 1.0 mL of ethyl acetate to obtain Suspension 1.

[0080] Step 2: Slowly add approximately 27 mg of hydrobromic acid (dissolved in 240 μL of ethanol) dropwise to Suspension 1, stir overnight at 4 °C until dissolved to obtain Solution 1.

[0081] Step 3: Add approximately 20 mL of n-heptane to Solution 1 to precipitate a solid, stir overnight at 4 °C to obtain Suspension 2.

[0082] Step 3: After standing, remove the supernatant, and vacuum dry the obtained solid overnight at room temperature to obtain the hydrobromide salt.

[0083] The product was characterized, and its XRPD pattern is shown in the appendix Figure 15 .

[0084] III. Comparison of the properties of the free base and different salts

[0085]

[0086] From the aspects and data examined in the above table, only the adipate salt is a crystalline salt form, and the crystal form preparation is controllable.

[0087] IV. Comparison of the pharmacokinetic characteristics of the free base and the crystalline form of the adipate salt

[0088] The present invention also studied the comparison of the pharmacokinetic parameters of the oral administration of the free base of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one and the hexanedioate crystalline form to Wistar rats. Each group of animals for the oral administration of the free base consisted of 4 animals, and each group of animals for the oral administration of the hexanedioate consisted of 3 animals, aged 6 - 8 weeks, male. The oral administration was 10% Cremophor EL + 90% (10% HP-β-CD in 1% HPMC (pH 2.2) in water). The animals in the oral administration group were fasted overnight and resumed feeding 4 hours after administration. The blood sampling points for the animals in the oral administration group were before administration and at 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration. Blood was collected from the jugular vein, and the blood volume collected at each sampling point was approximately 150 μL, anticoagulated with EDTA-K2, centrifuged at 2000 g for 5 min at 4°C within 15 minutes after sampling, and analyzed by LCMSMS-28 (Triple Quad 6500+).

[0089] The results showed that, compared with the free base, the T max was significantly prolonged. See the following table for details:

[0090]

[0091]

Claims

1. A crystalline form of the adipate salt of the (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one compound, characterized in that, The described crystalline form shows peaks at diffraction angles 2θ of 5.09°, 12.26°, 10.31°, 20.41°, 27.02°, 6.90°, 21.67°, 14.62°, 8.98°, 18.92°, 13.63°, 3.46°, 7.26°, 15.15°, 17.59°, 10.67° (±0.2°) on the X-ray diffraction pattern.

2. The crystalline form according to claim 1, wherein, The described crystalline form further includes peaks at diffraction angles 2θ of 4.77°, 22.83°, 28.99°, 23.13°, 19.96°, 16.98°, 14.06°, 27.84°, 18.18°, 16.31°, 25.48°, 24.68°, 21.30°, 26.51°, 30.38°, 19.35°, 11.75° (±0.2°).

3. The crystalline form according to claim 1 or 2, characterized in that Shows an X-ray diffraction pattern substantially as shown in Figure 5.

4. The preparation method of the crystalline form according to any one of claims 1-3, characterized in that Comprises the following steps: Step (1): Weigh a certain mass of the (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one free base compound and add it to a certain volume of ethyl acetate to obtain suspension 1. Step (2): Weigh a certain mass of the adipic acid ligand, ultrasonically heat and dissolve it in a certain volume of ethanol to obtain solution 1. Step (3): Dropwise add the solution 1 obtained in step (2) to the suspension 1 obtained in step (1) under stirring until it becomes clear, stir overnight at low temperature, and no solid is precipitated to obtain solution 2. Step (4): Add an anti-solvent to the solution 2 obtained in step (3), precipitate a solid, and stir at low temperature to obtain suspension 2. Step (5): Centrifuge the suspension 2 obtained in step 4, and vacuum dry the obtained solid at room temperature to obtain the described crystalline form.

5. The method according to claim 4, wherein, In the step (1), the molar ratio of the free base compound to the adipic acid ligand in step (2) is 1:1 - 1.

5.

6. The method according to claim 4, wherein, In the step (1), the molar volume ratio of the free base compound to ethyl acetate is: 1 - 10 mol: 0.1 - 1 L.

7. The method according to claim 4, wherein In the step (2), the molar volume ratio of the adipic acid ligand to ethanol is 1 - 10 mol: 0.1 - 1 L.

8. The method according to claim 4, wherein, The anti-solvent in the step (4) is selected from n-heptane.

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