WO2023125419A1 - Adipate crystal and preparation method therefor - Google Patents

Abstract

Provided are an adipate crystal of a PI3Kδ/γ dual inhibitor compound (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromene-4-one compound and a preparation method therefor. The crystal is more excellent in terms of physical properties and pharmacodynamics/pharmacokinetics.

Description

A kind of adipate crystal and preparation method thereof

This application claims the priority of the following Chinese patent applications: 1) Submitted to the State Intellectual Property Office of China on December 31, 2021, the application number is 202111679201.5, and the invention title is "A kind of adipate crystal and its preparation method" Chinese patent application; 2) A Chinese patent application submitted to the State Intellectual Property Office of China on March 11, 2022 with the application number 202210245009.3 and the title of the invention "A kind of adipate crystal and its preparation method". The entire contents of the above-mentioned Chinese patent applications are incorporated in this application by reference.

technical field

The present invention relates to the field of medicine, in particular to a PI3Kδ/γ dual inhibitor compound (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl) -Adipate crystals of 4H-chromen-4-one compounds and methods for their preparation.

Background technique

Phosphoinositide-3 kinases (PI3Ks) belong to a class of intracellular lipid kinases that phosphorylate the 3-hydroxyl of the inositol ring of phosphoinositide lipids (PI), thereby generating lipid second messengers. It has been reported in the art that targeted inhibitors of the phosphoinositide-3-kinase (PI3K) pathway can be used as immunomodulators.

A major issue 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 crystalline polymorph may change during manufacturing and/or storage, leading to quality control issues and formulation irregularities. Such variations may affect the reproducibility of the manufacturing process, resulting in a final formulation that does not meet the high quality and stringent requirements for formulation of pharmaceutical compositions. In this regard, it should generally be noted that any modification of the solid state of a pharmaceutical composition that improves its physical and chemical stability confers a significant advantage in stability relative to less stable forms of the same drug. Furthermore, it is crucial to develop stable production methods that consistently produce active substances. The existence of multiple crystalline forms with similar solubility poses a difficult challenge in the large-scale manufacture of pharmaceutical compounds.

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 polymorphs of a given substance have the same chemical composition, they can differ in one or more physical properties such as solubility, degree of dissociation, true density, dissolution, melting point, crystal shape, compaction behavior, flow properties and and/or differ from each other in terms of solid-state stability.

As generally stated above, polymorphic behavior of drugs can be of great importance in pharmacology. Differences in physical properties exhibited by polymorphs affect practical parameters such as storage stability, compressibility and density (important in formulation and product manufacturing) and dissolution rate (an important factor in determining bioavailability). Changes in chemical reactivity (e.g., differential oxidation, causing color to change more quickly when the dosage form is one polymorph than when the dosage form is another polymorph), mechanical changes (e.g., tablet discoloration after storage) Kinetically favorable polymorphs convert to thermodynamically more stable polymorphs) or both (e.g. tablets of one polymorph disintegrate more easily at high humidity) may lead to loss of stability difference. Additionally, the physical properties of the crystals may be important in processing. For example, one polymorph may be more likely to form solvates, leading to aggregation of the solid form and making handling of the solid more difficult. Alternatively, the particle shape and size distribution of one polymorph relative to another may differ, leading to increased challenges in filtering pharmaceutical actives to remove impurities.

While pharmaceutical formulations are expected to have improved chemical and physical properties, there is no predictable means to prepare new pharmaceutical forms (eg, polymorphs and other new crystalline forms) of existing molecules for such formulations. These new forms will provide consistency in physical properties across a range of environments common in fabrication and compositional use.

WO2014195888A1 patent document discloses (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromene-4-one compound, its structural formula It is formula I, and as a free base, it exhibits dual inhibitory functions of PI3Kδ/γ.

However, it is still urgently desired to develop a combination with (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one Compared with PI3Kδ/γ dual inhibitors, which have superior properties in terms of physical properties and pharmacodynamics/pharmacokinetics, and have higher applicability as pharmaceuticals.

Contents of the invention

The invention provides a hexane of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromene-4-one compound A crystalline form of a diacid salt, characterized in that the crystalline form shows 5.09°, 12.26°, 10.31°, 20.41°, 27.02°, 6.90°, 21.67°, 14.62°, 8.98°, Peaks at diffraction angles 2θ of 18.92°, 13.63°, 3.46°, 7.26°, 15.15°, 17.59°, 10.67° (±0.2°).

Further, the crystalline form also includes crystals at 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°, Peaks at diffraction angles 2θ of 26.51°, 30.38°, 19.35°, 11.75° (±0.2°).

Further, said crystalline form exhibits an X-ray diffraction pattern substantially as shown in FIG. 5 .

In addition, the present invention also provides a (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromene- Process for the preparation of crystalline forms of adipate salts of 4-keto compounds, characterized in that it comprises the following steps:

Step (1): Weigh a certain mass of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromene-4- A ketone free base compound is added to a certain volume of ethyl acetate to obtain a suspension 1;

Step (2): Weighing a certain amount of adipic acid ligand, ultrasonically heating and dissolving it in a certain volume of ethanol to obtain solution 1;

Step (3): Add the solution 1 obtained in step (2) dropwise to the suspension 1 obtained in step (1) which is being stirred, dissolve it, stir overnight at low temperature, no solid is precipitated, and obtain solution 2;

Step (4): adding an antisolvent to the solution 2 obtained in step (3) to precipitate a solid, stirring at a low temperature to obtain a suspension 2;

Step (5): The suspension 2 obtained in Step 4 was centrifuged, and the obtained solid was vacuum-dried at room temperature to obtain the crystalline form.

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

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

In the preferred technical solution of the present invention, wherein, the molar volume ratio of the adipic acid ligand to ethanol in the step (2) is 1-10mol:0.1-1L.

Description of drawings

The XRPD figure of accompanying drawing 1 free base compound;

The TGA figure of accompanying drawing 2 free alkali compounds;

The DSC figure of accompanying drawing 3 free alkali compounds;

The ¹ HNMR figure of accompanying drawing 4 free base compound;

The XRPD figure of accompanying drawing 5 adipate crystalline forms;

The TGA figure of accompanying drawing 6 adipate crystalline forms;

The DSC figure of accompanying drawing 7 adipate crystalline forms;

DVS figure and isotherm adsorption curve of accompanying drawing 8 adipate crystalline forms;

The PLM figure of accompanying drawing 9 adipate crystalline forms;

The ¹ HNMR figure of accompanying drawing 10 adipate crystal form;

The XRPD pattern of accompanying drawing 11 hemisuccinates;

The ¹ HNMR figure of accompanying drawing 12 hemisuccinates;

The XRPD pattern of accompanying drawing 13 phosphates;

The XRPD figure of accompanying drawing 14 oxalates;

The XRPD figure of accompanying drawing 15 hydrobromide;

Detailed ways

1. Analysis method

1.1 X-ray powder diffractometer (XRPD)

The crystal form of the samples was analyzed by X-powder diffractometer. The 2θ scanning angle of the sample is from 3° to 40°, the scanning step is 0.02°, and the scanning time of each step is 0.2s. The light tube voltage and current were 40kV and 40mA, respectively. When preparing samples, place an appropriate amount of samples on the sample tray to ensure that the surface of the samples is smooth and flat.

1.2 Differential Scanning Calorimetry (DSC)

The samples were analyzed by TA instruments Q200 DSC. Put the weighed sample (0.5mg-5mg) into the sample tray, and raise the sample to the final temperature at a rate of 10°C/min under the protection of nitrogen (50mL/min).

1.3 Thermogravimetric Analysis (TGA)

The samples were analyzed by TA instruments Q500. The sample was put into a tared platinum crucible, the system automatically weighed, and then the sample was raised to the final temperature at a rate of 10°C/min under the protection of nitrogen (40mL/min).

1.4 Polarized Light Microscopy (PLM)

The samples were analyzed using a polarizing microscope, and the morphology and microstructure of the crystals were obtained by adjusting different magnifications.

1.5 Dynamic moisture adsorption (DVS)

Dynamic moisture adsorption was performed using TA Instruments Q5000 SA. Approximately 1-10 mg of sample is placed in a sample pan and suspended from the sample chamber. The room temperature was maintained at a constant 25±1°C by a water bath. In the step mode, the sample is subjected to a cycle test in the relative humidity of 0%RH-80%RH. Analysis was performed at 10% RH/step. Set the time to maintain each humidity to 90min, so that the sample and the indoor environment can reach equilibrium.

1.6 Liquid hydrogen nuclear magnetic spectrum ( ¹ H NMR)

The samples were analyzed using Bruker Ascend 500MH, and the solvent was deuterated dimethyl sulfoxide.

2. Preparation method

2.1 The preparation method of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromene-4-one free base compound

Prepare (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one with reference to the method of patent document CN105358560A embodiment Free base compound, wherein, the XRPD spectrum of the free base compound is shown in Figure 1, the TGA spectrum is shown in Figure 2, the DSC spectrum is shown in Figure 3, and ^{the 1} HNMR spectrum is shown in Figure 4.

2.2 Preparation of crystalline form of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one adipate

Step 1: about 100 mg of the free base compound prepared in Example 2.1 was added to 2.0 mL of ethyl acetate to obtain a suspension 1;

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

Step 3: Add Solution 1 dropwise to Suspension 1 under stirring, dissolve it, stir overnight at 4°C, no solid precipitates out, and obtain Solution 2;

Step 4: Add about 20mL of n-heptane to solution 2, precipitate solid, stir at 4°C for about 8h to obtain a suspension;

Step 5: The suspension was centrifuged, and the resulting solid was vacuum dried overnight at room temperature to obtain the adipate salt.

Characterize the product, its XRPD spectrum is shown in Figure 5, TGA and DSC spectrum are shown in Figure 6 and Figure 7 respectively, DVS is shown in Figure 8, PLM is shown in Figure 9, ¹ HNMR As shown in Figure 10.

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

Step 1: About 50 mg of the free base compound prepared in Example 2.1 was added to 0.5 mL of isopropanol, and dissolved by heating in a water bath at 60°C to obtain solution 1;

Step 2: About 16mg of succinic acid was added to 0.2mL of isopropanol to obtain suspension 1;

Step 3: At room temperature, add the suspension 1 dropwise to the solution 1, dissolve it, stir overnight at 4°C, no solid is precipitated, and the solution 2 is obtained;

Step 4: Add about 5mL of n-heptane to solution 2, a solid precipitates, and stir overnight at room temperature to obtain suspension 2;

Step 5: Suspension 2 was centrifuged and the resulting solid was vacuum dried overnight at room temperature to yield the hemisuccinate.

The product was characterized, and its XRPD spectrum is shown in Figure 11, and its ¹ HNMR is shown in Figure 12.

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

Step 1: About 50 mg of the free base compound prepared in Example 2.1 was added to 0.5 mL of isopropanol, and dissolved by heating in a water bath at 60°C to obtain solution 1;

Step 2: About 16 mg of phosphoric acid (dissolved in 132 μL of isopropanol) was slowly dropped into solution 1 to precipitate a solid, and stirred overnight at 4°C to obtain a suspension;

Step 3: The suspension was centrifuged and the resulting solid was vacuum dried overnight at room temperature to yield the phosphate salt.

The product was characterized, and its XRPD spectrum is shown in Figure 13.

2.5 Preparation of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromene-4-one oxalate

Step 1: About 50 mg of the free base compound prepared in Example 2.1 was added to 0.5 mL of isopropanol, and heated in a water bath at 60°C to dissolve to obtain solution 1;

Step 2: About 17mg of oxalic acid, add 0.2mL of ethanol to obtain solution 2;

Step 3: Add Solution 2 dropwise to Solution 1 at room temperature, stir overnight at 4°C, and a solid precipitates out to obtain a suspension;

Step 4: The suspension was centrifuged, and the resulting solid was vacuum dried overnight at room temperature to obtain oxalate.

The product was characterized, and its XRPD spectrum is shown in Figure 14.

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

Step 1: About 50 mg of the free base compound prepared in Example 2.1 was added to 1.0 mL of ethyl acetate to obtain a suspension 1;

Step 2: About 27 mg of hydrobromic acid (dissolved in 240 μL ethanol), slowly drop into suspension 1, stir overnight at 4°C to dissolve, and obtain solution 1;

Step 3: Add about 20 mL of n-heptane to solution 1, precipitate solid, stir overnight at 4°C to obtain suspension 2;

Step 3: After standing still, the supernatant was removed, and the obtained solid was vacuum-dried at room temperature overnight to obtain hydrobromide.

The product was characterized, and its XRPD spectrum is shown in Figure 15.

3. Comparison of properties of free base and different salts

Table 1. Comparison of properties of free base and different salts

From the survey aspects and data in the above table, only adipate is in the crystalline salt form, and the preparation of the crystal form is controllable.

4. Comparison of pharmacokinetic characteristics of free base and crystalline form of adipate

The present invention also studies (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one free base and hexyl Comparison of pharmacokinetic parameters of Wistar rats administered orally in crystalline form of dibasic acid salt, 4 animals for each group of oral administration of free base, 3 animals for each group of oral administration of adipate, 6 -8 weeks old, male. Oral administration of 10% Cremophor EL+90% (10% HP-β-CD in 1% HPMC (pH 2.2) in water, the animals in the oral administration group fasted overnight, and resumed eating after 4 hours of administration. Oral administration group Animal blood collection points are before and after administration and 0.25, 0.5, 1, 2, 4, 8 and 24 hours. Jugular vein blood collection, the blood collection volume of each blood collection point is about 150 μ L, EDTA-K2 anticoagulant, within 15 minutes after sampling 4 Centrifuge at 2000g for 5min, and analyze by LCMSMS-28 (Triple Quad 6500+).

It was found that the _Tmax of the crystalline form of adipate was significantly prolonged compared to the free base. See the table below for details:

Table 2. Comparison of Pharmacokinetic Profiles of Free Base and Crystalline Adipate Salt Forms

Claims (8)

1. A kind of adipate salt of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one compound A crystalline form, characterized in that the crystalline form shows 5.09°, 12.26°, 10.31°, 20.41°, 27.02°, 6.90°, 21.67°, 14.62°, 8.98°, 18.92°, 13.63° on the X-ray diffraction pattern °, 3.46°, 7.26°, 15.15°, 17.59°, 10.67°, (±0.2°) peaks at diffraction angles 2θ.
2. The crystalline form according to claim 1, wherein the crystalline form also includes crystalline forms at 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°) at the diffraction angle 2θ.
3. The crystalline form according to claim 1 or 2, characterized by exhibiting an X-ray diffraction pattern substantially as shown in FIG. 5 .
4. A process for the preparation of a crystalline form according to any one of claims 1-3, characterized in that it comprises the following steps:

   Step (1): Weigh a certain mass of (S)-2-(1-(9H-purin-6-ylamino)propyl)-3-(3-fluorophenyl)-4H-chromene-4- A ketone free base compound is added to a certain volume of ethyl acetate to obtain a suspension 1;

   Step (2): Weighing a certain amount of adipic acid ligand, ultrasonically heating and dissolving it in a certain volume of ethanol to obtain solution 1;

   Step (3): Add the solution 1 obtained in step (2) dropwise to the suspension 1 obtained in step (1) which is being stirred, dissolve it, stir overnight at low temperature, no solid is precipitated, and obtain solution 2;

   Step (4): adding an antisolvent to the solution 2 obtained in step (3) to precipitate a solid, stirring at a low temperature to obtain a suspension 2;

   Step (5): The suspension 2 obtained in Step 4 was centrifuged, and the obtained solid was vacuum-dried at room temperature to obtain the crystalline form.
5. The method according to claim 4, wherein the molar ratio of the free base compound described in the step (1) to the adipic acid ligand described in the 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: 1-10mol: 0.1-1L.
7. The method according to claim 4, wherein the molar volume ratio of the adipic acid ligand to ethanol in the step (2) is 1-10mol:0.1-1L.
8. The method according to claim 4, wherein, the antisolvent in the described step (4) is selected from n-heptane.

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