WO2022144042A1 - Crystal form of tas-116, and preparation method therefor, pharmaceutical composition, and use therefor - Google Patents

Abstract

Disclosed are a crystal form of TAS-116, and a preparation method therefor, pharmaceutical composition, and use therefor.

Description

Crystal form of TAS-116 and its preparation method, pharmaceutical composition and use

Citations to Related Applications

This disclosure requires Chinese Invention Patent Application No. 202011630937.9, filed with the State Intellectual Property Office of the People's Republic of China on December 31, 2020, and Chinese Invention Patent Application No. 202111665564.3, filed with the State Intellectual Property Office of the People's Republic of China on December 31, 2021 and the entire contents of which are incorporated into this disclosure by reference.

field

The present disclosure relates to the field of medicinal chemistry. In particular, the present disclosure relates to crystalline forms of TAS-116 and methods for their preparation, pharmaceutical compositions and uses.

background

TAS-116 is a selective heat shock protein 90 (HSP90) inhibitor for the treatment of gastrointestinal cancers such as colon cancer and gastric stromal tumor. Currently, TAS-116 is undergoing clinical studies in patients with gastrointestinal cancers.

The chemical name for TAS-116 is 3-ethyl-4-{3-isopropyl-4-(4-(1-methyl-1H-pyrazol-4-yl)-1H-imidazol-1-yl)- 1H-pyrazolo[3,4-b]pyridin-1-yl}benzamide, also known as THS-1593, molecular formula is C ₂₅ H ₂₆ N ₈ O, molecular weight is 454.53, and its chemical structure is as follows:

Patent CN102471335B discloses the general structure of compounds including TAS-116.

Patent CN104710420B discloses the specific structure of TAS-116, and discloses a preparation method of free state of TAS-116, mentioning that TAS-116 is a white solid.

Patent CN107531707B discloses type I crystals (hereinafter referred to as "crystal form I") and type II crystals (hereinafter referred to as "crystal form II") of TAS-116, wherein crystal form I only has an XRPD spectrum, and the oral administration of crystal form I Absorption problems exist; Form II is anhydrous, and the patent also informs that Form II is a better form than Form I.

During the research process, the inventor found that the sample of TAS-116 prepared according to Example 102 in the patent CN104710420B was the crystal form II, and the DMSO (dimethyl sulfoxide) residue of the obtained sample was very high. more residue. The inventors found that the crystal form II sample appeared in the crystal form I after being retained in the solvent of Example 102 for more than 2 hours, indicating that the stability of the crystal form II was problematic.

The inventors also found in the research process that the TAS-116 crystal form I prepared according to CN107531707B is unstable, and is prone to crystal form transformation when placed at room temperature (20-30° C.), and transforms into crystal form II. Form I requires more severe storage conditions to be stable, indicating that its medicinal value is not high.

The inventors also found in the research process that the stability of TAS-116 crystal form II prepared according to the CN107531707B patent has great limitations. Crystallization also occurred in the environment, and the crystal form I appeared in different degrees.

In view of the shortcomings of the existing technology, especially the problem of insufficient stability, it is necessary in the art to develop a more stable solid form of TAS-116 with more advantageous properties.

Overview

In view of the deficiencies of the prior art, the purpose of the present disclosure is to provide a TAS-116 crystal form with better physicochemical properties, as well as a preparation method, pharmaceutical composition and use thereof. Compared with the known TAS-116 compound, the TAS-116 crystal form of the present disclosure has better crystal form stability. In addition, it was found to have other unexpected effects, mainly manifested in better solubility and dissolution rate, better particle morphology, higher crystallinity, better hygroscopicity, etc. It is further considered that the present disclosure The crystalline form also possesses better fluidity, formulation processability and bioavailability.

According to the purpose of the present disclosure, a first aspect of the present disclosure provides TAS-116 crystalline form, hereinafter referred to as Form III.

The crystal form III of the present disclosure is an anhydrate, and its structural formula is shown in formula (I):

Using Cu-Kα radiation, the X-ray powder diffraction pattern of the crystal form III has characteristic peaks at 2θ values of 6.39°±0.2°, 10.40°±0.2°, 12.94°±0.2° and 19.48°±0.2° .

In certain embodiments, using Cu-Kα radiation, the X-ray powder diffraction pattern of said Form III has at least 2θ values of 3.20°±0.2°, 6.39°±0.2°, 10.40°±0.2°, 11.41 There is a characteristic peak at one of °±0.2°, 12.94°±0.2° and 19.48°±0.2°.

In certain embodiments, using Cu-Kα radiation, the X-ray powder diffraction pattern of said Form III has at least 2θ values of 3.20°±0.2°, 6.39°±0.2°, 10.40°±0.2°, 11.41 One of °±0.2°, 12.94°±0.2°, 17.58°±0.2°, 18.26°±0.2°, 19.48°±0.2°, 21.71°±0.2°, 22.69°±0.2° and 23.65°±0.2° has characteristic peaks.

In certain embodiments, using Cu-Kα radiation, the X-ray powder diffraction pattern of Form III has 2θ values of 3.20°±0.2°, 6.39°±0.2°, 10.40°±0.2°, 11.41° There are characteristic peaks at ±0.2°, 12.94°±0.2° and 19.48°±0.2°.

In certain embodiments, using Cu-Kα radiation, the X-ray powder diffraction pattern of Form III has 2θ values of 3.20°±0.2°, 6.39°±0.2°, 10.40°±0.2°, 11.41° There are characteristic peaks at ±0.2°, 12.94°±0.2°, 17.58°±0.2°, 18.26°±0.2°, 19.48°±0.2°, 21.71°±0.2°, 22.69°±0.2° and 23.65°±0.2°.

In certain embodiments, using Cu-Kα radiation, the X-ray powder diffraction pattern of Form III has characteristic peaks at the following diffraction angles with 2θ values and relative intensities as shown in the table below:

In certain embodiments, the X-ray powder diffraction (XRPD) pattern of the Form III is substantially as shown in FIG. 4 .

In certain embodiments, the differential scanning calorimetry (DSC) pattern of the Form III is shown in FIG. 5 . The crystal form III has a melting point onset value of 269°C and a peak value of 270°C.

In certain embodiments, the thermogravimetric analysis (TGA) pattern of the Form III is shown in FIG. 6 . The crystal form III is anhydrous, with only 0.9% weight loss before 105°C.

In certain embodiments, the Fourier transform infrared spectrum of the Form III is shown in FIG. 7 . The Fourier transform infrared spectrum of the crystal form III is at wavenumbers of 1650cm- ¹ ±2cm- ¹ , 1569cm- ¹ ±2cm- ¹ , 1504cm ^-1 ±2cm ^-1 , 1424cm ^-1 ±2cm ^-1 , 1272cm ^-1 ±1 2cm ^-1 , 1030cm ^-1 ±2cm ^-1 , 823cm ^-1 ±2cm ^-1 , 812cm ^-1 ±2cm ^-1 , 748cm ^-1 ±2cm ^-1 , 711cm ^-1 ±2cm ^{-1 ,} 668cm ^-1 ±2cm ^- There are characteristic peaks at ¹ and 653 cm ^-1 ± 2 cm ^-1 .

In certain embodiments, the DVS isotherm of Form III is shown in FIG. 8 . The crystal form III has only a 0.3% weight change in the humidity range of 0%RH-80%RH.

In certain embodiments, a polarized light microscope (PLM) image of the Form III is shown in FIG. 9 . The crystal form III is a fine granular crystal.

According to the purpose of the present disclosure, the second aspect of the present disclosure provides a preparation method of TAS-116 crystal form III, including any one of the following methods:

(1) The solid compound TAS-116 was formed into a suspension in solvent 1, stirred, the solid was separated, and dried to obtain the crystal form III.

In certain embodiments, the solvent 1 is selected from cyclic ethers. In certain embodiments, the solvent 1 is 1,4-dioxane.

In certain embodiments, the mass volume ratio of the compound TAS-116 solid to solvent 1 is 10 to 200:1 (mg:mL). In certain embodiments, the mass volume ratio of the compound TAS-116 solid to solvent 1 is 30 to 100:1 (mg:mL).

In certain embodiments, the stirring time is from 10 hours to 168 hours. In certain embodiments, the stirring time is from 16 hours to 72 hours.

In certain embodiments, the stirring is performed at room temperature.

In certain embodiments, the resulting solid is reheated at a temperature of 60°C to 150°C after drying. In certain embodiments, the resulting solid is reheated at a temperature of 80°C to 130°C after drying. In certain embodiments, the heating time is 5 minutes to 16 hours.

(2) The solid compound TAS-116 is formed into a solution in solvent 2, the temperature is lowered, the solid is separated, and dried to obtain crystal form III.

In certain embodiments, the solvent 2 is selected from halogenated alkanes, alcohols, ketones, furans, cyclic ethers, nitriles or mixed solvents thereof. In certain embodiments, the solvent 2 is tetrahydrofuran and chloroform. In certain embodiments, the volume ratio of the two solvents in the mixed solvent is 1:4 to 4:1.

In certain embodiments, the mass volume ratio of solute to solvent 2 in the solution is 5 to 100:1 (mg:mL). In certain embodiments, the mass volume ratio of solute to solvent 2 in the solution is 40 to 100:1 (mg:mL).

In certain embodiments, the temperature at which the solution is formed is 60°C to 80°C.

In certain embodiments, the temperature is lowered to 10°C to 50°C. In certain embodiments, the temperature is lowered to 10°C to 40°C.

In certain embodiments, Form III seeds of TAS-116 are added during the cooling process.

In certain embodiments, the seed crystals are added in an amount ranging from 10% to 30% TAS-116 solids.

In certain embodiments, the cooling may be concurrent with stirring or cooling followed by stirring. In certain embodiments, the stirring time is from 2 hours to 24 hours.

Compared with the known crystal forms, the TAS-116 crystal form III of the present disclosure has the following advantages:

In certain embodiments, the TAS-116 crystalline form III of the present disclosure has better stability. Compared with the known TAS-116 crystal form I, it has better crystal form stability under thermal conditions; compared with the known TAS-116 crystal form II, it has better crystal form stability under water conditions The range of water conditions includes but is not limited to aqueous solutions, aqueous media, body fluids and humidity environments, etc.; Form I cannot tolerate heat (40°C), and Form II cannot tolerate water, and neither can maintain the original There are crystalline forms. Thermal conditions and water are very common in the process of preparing API, during the drying stage, during the preparation operation, during the shelf life of the drug, during the disintegration and dissolution in vivo, so the crystal form 1 and crystal form II of TAS-116 will inevitably be In the above process, the crystal form is unstable.

It is well known that the transformation of the crystal form will adversely affect the dissolution, dissolution, bioavailability, stability, safety, compliance, etc. of the drug. Therefore, it is necessary to avoid the use of crystal form I and crystal form II, and crystal form III is more medicinal value.

In certain embodiments, the TAS-116 Form III of the present disclosure has a faster dissolution rate than the known TAS-116 Form I and Form II.

In certain embodiments, the TAS-116 Form III of the present disclosure has a higher solubility than the known TAS-116 Form I and Form II.

In certain embodiments, the TAS-116 crystalline form III of the present disclosure has a moisture absorption of 0.3% in a 0% to 80% RH environment, and is not easily hygroscopic.

The TAS-116 crystal form III of the present disclosure is a uniform and fine powder, which has better solubility and dissolution rate, which is beneficial to improve the bioavailability of the drug; has better particle size distribution, and is easier to achieve mixing uniformity and content uniformity; It has lower hygroscopicity and is more stable in humidity environment.

In the preparation method of TAS-116 crystal form III of the present disclosure:

Unless otherwise noted, "overnight" means 10 hours to 16 hours.

"Room temperature" refers to a temperature of 10 to 30°C.

For "stirring", conventional methods in the art can be used, for example, stirring methods include magnetic stirring and mechanical stirring, and the stirring speed is 50 to 1800 rpm. In certain embodiments, the stirring speed is 300 to 900 rpm.

"Isolation" can be performed by conventional methods in the art, such as centrifugation or filtration. In certain embodiments, filtration under reduced pressure, typically suction filtration, is performed at room temperature at less than atmospheric pressure. In certain embodiments, the pressure is less than 0.09 MPa.

"Drying" can be accomplished by using conventional techniques in the art, such as drying at room temperature, air drying or drying under reduced pressure; it can be under reduced pressure or normal pressure, preferably the pressure is less than 0.09 MPa. The drying apparatus and method are not limited, and can be a fume hood, a forced air oven, a spray dryer, a fluidized bed drying or a vacuum oven; it can be carried out under reduced or no reduced pressure, preferably the pressure is less than 0.09Mpa. The starting material TAS-116 can be prepared by referring to the method described in Example 102 in the patent document CN104710420B, or it can be purchased from the market, which is incorporated into the present disclosure by reference in its entirety.

In the present disclosure, "crystalline form" means as evidenced by the characterization of the X-ray powder diffraction pattern shown. It is well known to those skilled in the art that the experimental errors therein depend upon instrumental conditions, sample preparation and sample purity. Spectra generally vary with instrument conditions. The relative intensities of peaks may vary with experimental conditions, so the order of peak intensities cannot be used as the only or decisive factor; experimental errors in peak angles should also be taken into account, usually allowing an error of ±0.2°; the influence of factors such as sample height can cause The peak angle is shifted as a whole, and a certain shift is usually allowed. Therefore, those skilled in the art can understand that any crystal form having the same or similar characteristic peaks as the X-ray powder diffraction pattern of the present disclosure belongs to the scope of the present disclosure. The "single crystal form" refers to a single crystal form detected by X-ray powder diffraction.

The novel crystalline form of TAS-116 of the present disclosure is pure, single, and substantially not mixed with any other crystalline form or amorphous state. In the present disclosure, "substantially absent" when used to refer to a new crystal form means that the new crystal form contains less than 20% by weight of other crystal forms or amorphous states, more particularly less than 10% by weight, especially It means less than 5% by weight, especially less than 1% by weight.

According to the purpose of the present disclosure, the third aspect of the present disclosure provides a pharmaceutical composition, the pharmaceutical composition comprises a therapeutically effective amount of the TAS-116 crystal form III or the TAS-116 crystal form III prepared by the method of the present disclosure, and at least one pharmaceutically acceptable carrier.

The pharmaceutical compositions provided by the present disclosure can be administered by a number of routes including, but not limited to: oral (enteral) administration, parenteral (injection) administration, rectal administration, topical administration, transdermal administration, intradermal administration administration, intrathecal, subcutaneous (SC), intramuscular (IM), sublingual/buccal, ocular, otic, vaginal and intranasal or inhalation, usually, an effective amount of The present disclosure provides solid forms of TAS116. The solid form of TAS116 actually administered can be determined by the physician depending on the circumstances, including the condition being treated, the route of administration chosen, the compound actually administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, etc. amount.

Oral compositions can take the form of bulk liquid solutions or suspensions or bulk powders. More usually, however, the compositions are provided in unit dosage form. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human patients and other mammals, each unit containing a predetermined quantity of active material suitable for producing the desired therapeutic effect in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, premeasured ampoules or syringes of liquid compositions, or, in the case of solid compositions, pills, tablets, capsules, etc., either conventional immediate release formulations or Dispersible, chewable, orally dissolving formulations, or sustained-release formulations, may be enteric-coated tablets.

For oral doses, a typical regimen is one to five oral doses per day, especially one to four oral doses. Using these dosing modes, each dose provides about 0.01 to about 20 mg/kg of the solid form of TAS-116 provided by the present disclosure, with preferred doses each providing about 0.1 to about 10 mg/kg, especially about 0.2 to about 5 mg /kg, which depends on the particular condition being treated, the age and weight of the particular patient, and the particular patient's response to drug therapy, the exact dose is to be determined under the guidance of a physician and in accordance with standard medical principles. A specific unit dose can be, for example, 20 mg, 40 mg, 60 mg, 80 mg, 100 mg, 120 mg, 140 mg, 160 mg, 200 mg, and the like.

The pharmaceutically acceptable carriers or adjuvants in the pharmaceutical composition are well known to those skilled in the art, and can have various well-known forms, such as, but not limited to, diluents, such as starch, modified starch, lactose, powder, etc. cellulose, microcrystalline cellulose, calcium hydrogen phosphate anhydrous, tricalcium phosphate, mannitol, sorbitol, sugar, etc.; binders, such as gum arabic, guar gum, gelatin, polyvinylpyrrolidone, hydroxypropyl cellulose Vinegar, hydroxypropyl methylcellulose, polyethylene glycol, copovidone, etc.; disintegrants, such as starch, sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, crospovidone, crospovidone Sodium bicarboxymethylcellulose, colloidal silicon dioxide, etc.; lubricants, such as stearic acid, magnesium stearate, zinc stearate, sodium benzoate, sodium acetate, etc.; glidants, such as colloidal silicon dioxide, etc.; Complex formers such as various grades of cyclodextrins and resins; release rate controlling agents such as hydroxypropyl cellulose, hydroxymethyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, methyl cellulose Vine, methyl methacrylate, wax, etc. Other pharmaceutically acceptable carriers or adjuvants that can be used include, but are not limited to, film formers, plasticizers, colorants, flavors, viscosity modifiers, preservatives, antioxidants, and the like. In the case of oral tablets, commonly used carriers include lactose and corn starch, and lubricants such as magnesium stearate may also be added; in the case of oral capsules, useful carriers/diluents include lactose, high and low molecular weight polyethylene glycols and dry corn starch; in the case of gelatin capsules, powdered carriers or adjuvants such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like; when administered orally as a suspension, The active ingredient is mixed with emulsifying and suspending agents; if desired, certain sweetening and/or flavoring and/or coloring agents may be added. Each carrier or adjuvant must be acceptable, compatible with the other ingredients in the formulation and not injurious to the patient.

According to the purpose of the present disclosure, the fourth aspect of the present disclosure provides the TAS-116 crystal form III, the TAS-116 crystal form III obtained by the preparation method of the present disclosure, or the pharmaceutical composition in the preparation of a drug for the treatment of cancer use in.

In certain embodiments, the cancer includes, but is not limited to, the following specific diseases and conditions, such as head and neck cancer, esophageal cancer, stomach cancer, colon cancer, rectal cancer, liver cancer, gallbladder-cholangiocarcinoma, biliary tract cancer, pancreas cancer Cancer, lung cancer, breast cancer, ovarian cancer, cervical cancer, endometrial cancer, kidney cancer, bladder cancer, prostate cancer, testicular tumor, bone-soft tissue sarcoma, leukemia, malignant lymphoma, multiple myeloma, skin cancer, brain cancer Tumors, mesothelioma, etc.

According to the purpose of the present disclosure, a fifth aspect of the present disclosure provides a method of treating cancer, the method comprising administering to a patient in need of the method a therapeutically effective amount of the TAS-116 Form III or a pharmaceutical composition thereof.

In certain embodiments, the patient includes, but is not limited to, a mammal.

According to the purpose of the present disclosure, the sixth aspect of the present disclosure provides the combined use of the TAS-116 crystal form III or its pharmaceutical composition with other drugs.

In certain embodiments, the other drug is a monoclonal antibody drug. In certain embodiments, the other drugs are PD-1 and PD-L1 inhibitors. In certain embodiments, the other drug is nivolumab, AB122, and the like.

Brief Description of Drawings

Figure 1 is the XRPD pattern (crystal form II) of the TAS-116 compound prepared according to the method described in Example 102 in the patent document CN104710420B.

Fig. 2 is the XRPD pattern of the TAS-116 compound crystal form I (type I crystal) prepared according to the method described in the comparative example 1 in the patent document CN107531707B.

Figure 3 is the XRPD pattern of the TAS-116 compound crystal form II (type II crystal) prepared according to the method described in Example 1 in the patent document CN107531707B.

FIG. 4 is the XRPD pattern of TAS-116 crystal form III prepared in Example 1 of the present disclosure.

FIG. 5 is the DSC spectrum of TAS-116 crystal form III prepared in Example 1 of the present disclosure.

6 is the TGA spectrum of TAS-116 crystal form III prepared in Example 1 of the present disclosure.

FIG. 7 is the IR spectrum of TAS-116 crystal form III prepared in Example 1 of the present disclosure.

FIG. 8 is the DVS spectrum of TAS-116 crystal form III prepared in Example 1 of the present disclosure.

9 is a PLM image of the TAS-116 crystal form III prepared in Example 1 of the present disclosure.

FIG. 10 is the XRPD pattern of TAS-116 crystal form I under thermal conditions in Experiment 1 of Experimental Example 1. FIG.

FIG. 11 is the XRPD pattern of TAS-116 crystal form II in water condition in Experiment 1, Experiment 1. FIG.

FIG. 12 is the XRPD pattern of TAS-116 crystal form II and crystal form III under thermal conditions in Experiment 1 of Experimental Example 1. FIG.

FIG. 13 is the XRPD pattern of TAS-116 crystal form I and crystal form III in water condition in Experiment 1, Experiment 1. FIG.

FIG. 14 is an XRPD comparison diagram of the crystal form stability test of TAS-116 crystal form III in Experiment 1, Experiment 2. FIG.

FIG. 15 is an XRPD comparison diagram of the 63-day stability test of the crystal form of TAS-116 crystal form III in Experiment 1, Experiment 2. FIG.

FIG. 16 is a particle size distribution (PSD) diagram of TAS-116 crystal form III in Experimental Example 2. FIG.

Figure 17 is the solubility curve of crystal form I, crystal form II and crystal form III in the buffer solution of pH 6.8 in Experiment 1 of Experimental Example 3.

18 is the tablet dissolution curves of TAS-116 crystal form I, crystal form II and crystal form III in Experimental Example 4.

detail

The following examples further illustrate the present disclosure, however, they are not intended to limit or limit the scope of the present disclosure. All patent documents and non-patent publications cited in this specification are incorporated by reference in their entirety.

Testing instruments and methods:

X-ray powder diffraction (XRPD): The instrument is a Bruker D8 Advance diffractometer. Samples were tested at room temperature. The detection conditions are as follows, angle range: 3 to 40° 2θ, step size: 0.02° 2θ, speed: 0.2 sec/step.

Polarized Light Microscopy (PLM) images were taken from an XP to 500E polarized light microscope. Take a small amount of powder sample and place it on a glass slide, add a small amount of mineral oil dropwise to disperse the sample, cover it with a cover glass, place it on the stage for observation and take pictures.

Thermogravimetric analysis (TGA) data were obtained from TA Instruments Q500 TGA. The detection method is as follows: take the sample and raise the sample to 400 °C at a heating rate of 10 °C/min under the protection of 40 mL/min of dry _N2 .

Dynamic moisture sorption (DVS) analysis data and isothermal adsorption analysis data were obtained from SMS Intrinsic PLUS. The detection method is as follows: take a sample and detect the weight change during the change of relative humidity from 0% to 80% to 0%.

Infrared spectroscopy (IR) data were collected from Bruker Tensor 27 using ATR equipment to collect infrared absorption spectra in the range of 600 to 4000 cm ⁻¹ .

HPLC purity data were obtained from a Uitimate 3000 high performance liquid chromatograph. The chromatographic column is C18 (4.6*150mm, 5μm), the detection wavelength is 254nm, the detection column temperature is 30°C, the flow rate is 1mL/min, and the injection volume is 5μL.

Unless otherwise specified, the examples are all operated at room temperature, and the solvent ratios are all volume ratios.

All reagents used in the examples are commercially available unless otherwise specified.

Preparation Example 1 Preparation of free state of TAS-116

According to the method described in Example 102 (P77) in the patent document CN104710420B, TAS-116 was prepared as a free white solid.

After testing, its XRPD pattern is shown in Figure 1, which shows that TAS-116 prepared according to the method described in Example 102 in the patent document CN104710420B is crystal form II.

Preparation Example 2 Preparation of TAS-116 Form I

The free state sample of TAS-116 was prepared according to the method described in Comparative Example 1 in the patent document CN107531707B.

After testing, its XRPD pattern is shown in Figure 2, indicating that the free state sample of TAS-116 prepared according to the method described in Comparative Example 1 in the patent document CN107531707B is crystal form I.

Preparation Example 3 Preparation of TAS-116 Form II

The free state sample of TAS-116 was prepared according to the method described in Example 1 in the patent document CN107531707B.

After testing, its XRPD pattern is shown in Figure 3, indicating that the free state sample of TAS-116 prepared according to the method described in Example 1 in the patent document CN107531707B is crystal form II.

Example 1 Preparation of TAS-116 crystal form III

About 500 mg of TAS-116 was taken, 10 mL of 1,4-dioxane was added to form a suspension, stirred at room temperature overnight, centrifuged, and vacuum dried at room temperature overnight to obtain a solid, and then heated at 120 ° C for 5 minutes to obtain about 401 mg of white solid.

After testing, the obtained solid is TAS-116 crystal form III, which

The XRPD pattern is shown in Figure 4;

The DSC spectrum is shown in Figure 5, the melting point onset value is 269 °C, and the peak value is 270 °C;

The TGA spectrum is shown in Figure 6; it is anhydrous, with only 0.9% weight loss before 105 °C;

The IR spectrum is shown in Figure 7;

The DVS diagram is shown in Fig. 8, with only 0.3% weight change in the humidity range of 0%RH-80%RH;

The PLM diagram is shown in Figure 9. The fine granular crystals have a crystal size far less than 50 microns, and the fine particles are more conducive to improving the solubility and dissolution rate.

Example 2 Preparation of TAS-116 crystal form III

About 1000 mg of TAS-116 was taken, 5 mL of 1,4-dioxane was added to form a suspension, stirred at room temperature for 72 hours, centrifuged, and vacuum dried at room temperature overnight to obtain a solid, and then heated at 150 °C for 5 minutes to obtain about 850 mg of white solid.

Example 3 Preparation of TAS-116 crystal form III

Take about 200 mg of TAS-116, add 20 mL of 1,4-dioxane to form a solution, which becomes a suspension after 0.5 hours, stir at room temperature for 10 hours, centrifuge, and vacuum dry at room temperature overnight to obtain a solid, which is then heated at 60°C for 16 hours About 100 mg of white solid were obtained afterward.

Example 4 Preparation of TAS-116 crystal form III

Take about 50 mg of TAS-116 prepared in Preparation Example 1, add 1.0 mL of chloroform, form a solution at 60 °C, cool down to 30 °C to 40 °C to obtain a saturated solution, add 5 mg of the crystal form III seed crystal prepared in Example 1, and stir for 2 hours, Continue to cool down to 0°C, centrifuge, and vacuum dry at room temperature overnight to obtain about 47 mg of white solid.

Example 5 Preparation of TAS-116 crystal form III

Take about 50 mg of TAS-116 prepared in Preparation Example 1, add 1.0 mL of chloroform and 0.25 mL of tetrahydrofuran, form a solution at 60°C, cool down to room temperature to obtain a saturated solution, add 15 mg of the crystal form III seed crystal prepared in Example 1, and stir for 24 hours, Centrifugation and vacuum drying overnight at room temperature yielded about 44 mg of a white solid.

After testing, the solids obtained in Examples 2 to 5 are TAS-116 crystal form III, which has the same characteristic pattern as the crystal form III prepared in Example 1, and is not repeated in this disclosure.

Experimental example 1 stability comparison experiment

experiment one

The disclosed crystal form I, crystal form II samples and crystal form III samples of the present disclosure were taken to investigate the crystal form changes after heating and under water conditions.

Thermal conditions: including but not limited to blast drying method above 30°C, vacuum drying method, acceleration (40°C/75%RH) influencing factors, heating method in solvent above 30°C, etc. The thermal conditions of the present disclosure refer to a 40°C blast drying method.

Aqueous conditions: including but not limited to samples in aqueous solutions, aqueous media, body fluids, and humidity. The water conditions of the present disclosure are stirring in an aqueous solution at room temperature for 16 hours.

The experimental results are shown in Table 1 below. Table 1 shows that the crystal form I cannot withstand heat, and the crystal form II cannot withstand water, and the original crystal form cannot be maintained under these two conditions. However, the crystal form III of the present disclosure has good thermal and water crystal form stability.

Table 1

Experiment 2

An appropriate amount of the crystal form III sample of the present disclosure was weighed and placed in the open under long-term (25°C/60%RH), accelerated (40°C/75%RH), and high temperature (40°C) conditions, and XRPD was regularly detected.

The experimental results show that the crystal form III has not been transformed for 15 days under long-term, accelerated and high-temperature conditions, and its XRPD comparison chart is shown in Figure 14.

Further, the crystal form III exposure remained unchanged for 63 days under long-term (25°C/60%RH) and high temperature (40°C) conditions, and its XRPD comparison chart is shown in Figure 15 .

Experiment 3

An appropriate amount of the crystal form III sample of the present disclosure was weighed and placed openly under long-term (25°C/60%RH), accelerated (40°C/75%RH), high temperature (40°C) and high humidity (97%RH) conditions , take the initial 0 days, long-term (25°C/60%RH) 63 days, accelerated (40°C/75%RH) 63 days, high temperature (40°C) 63 days and high humidity (97%RH) 63 days Samples of Form III were tested for purity by HPLC.

The HPLC detection method is shown in Table 2 below.

Table 2

Category	parameter
instrument	Uitimate3000 High Performance Liquid Chromatograph
Column Type	C18,4.6×150mm,5um
UV wavelength	UVat254nm
flow rate	1.0mL/min
column temperature	30℃
Injection volume	5.0uL
thinner	methanol
mobile phase A	Water (0.05%FA)
mobile phase B	Methanol (0.05%FA)
mobile phase ratio	A:B=45:55

The purity test results of crystal form III are shown in Table 3 below. The purity test results in Table 3 show that the crystal form III of the present disclosure has good chemical stability.

table 3

Experimental Example 2 Particle Size Distribution

An appropriate amount of samples of the crystal form III of the present disclosure were respectively taken, dispersed with n-heptane, and then detected by a laser particle size analyzer. The particle size distribution (PSD) was collected on a Microtrac Model S3500 laser particle size analyzer. The method parameters are as follows in Table 4:

Table 4

parameter	Numerical value
Zero time	30s
Detection time	30s
Dispersant flow rate	55%
Dispersant	n-heptane
Laser source wavelength	780nm

The experimental results are shown in Table 5 below and FIG. 16 . Figure 16 shows that the PSD of Form III is normally distributed.

table 5

Crystal form	D (10) (μm)	D (50) (μm)	D (90) (μm)
Form III	5.42	10.75	25.47

Experimental Example 3 Solubility Test

Experiment 1 Solubility at pH 6.8

Take about 10 mg of each of crystal form I, crystal form II and crystal form III in a bottle, add 15 mL of pH 6.8 buffer to form a suspension, then shake all samples at 25 °C ± 2 °C, respectively At 5min, 30min, 45min, and 60min, 1 mL of each crystal form sample in the pH 6.8 buffer was taken and filtered, and the solubility of the sample was detected by HPLC. The HPLC detection method is the same as that in Table 2.

Buffer solution with pH=6.8: prepare 100 mL of 0.2 mol/L potassium dihydrogen phosphate, and adjust the pH to 6.8 with 0.2 mol/L sodium hydroxide.

The solubility test results of crystal form I, crystal form II and crystal form III in the buffer solution of pH 6.8 are shown in Table 6 below, and the solubility curve is shown in Figure 17.

Table 6

Solubility in experimental water

Take a sample of about 4 mg of crystal form III in a bottle, add 15 mL of water to form a suspension, then shake at 25 ° C ± 2 ° C, take 1 mL of the sample after shaking for 60 min, filter, and detect by HPLC (the same method as Table 6) Sample solubility, the solubility was 5.83 μg/mL.

Experimental Example 4 Tablet Dissolution

Tablet production and dissolution test process:

Take an appropriate amount of the crystal form I, crystal form II and crystal form III samples in the following table 7 and mix them evenly.

Table 7 Tablet prescription table

The mixed samples of the raw and auxiliary materials of the three crystal forms were tableted with a single punch tablet machine (pressure 2MPa, time 2min).

Dissolution medium: pH 6.8 phosphate buffer.

Dissolution tests were performed on the tablets containing Form I, Form II and Form III, respectively.

Dissolution method: temperature 37°C, rotating speed 50rpm, medium volume 900mL, slurry method. (Dissolution Apparatus: Agilent 708-DS, Sampler: Agilent 850-DS)

Samples were taken at nine time points, 5min, 10min, 15min, 20min, 30min, 45min, 1h, 1.5h and 2h, respectively.

Experimental results:

In the same time, the dissolution rate of crystal form III was higher than that of crystal form I and crystal form II; after 2 h, the dissolution rate of crystal form III reached 49%, while the dissolution rates of crystal forms I and II were only 29% and 33%. The tablet dissolution curves of crystal form I, crystal form II and crystal form III are shown in Figure 18, and the specific dissolution test results of crystal form I, crystal form II and crystal form III are shown in Table 8 below.

Table 8

The above are only specific embodiments of the present disclosure, but the protection scope of the present disclosure is not limited thereto. Any person skilled in the art may, within the technical scope disclosed by the present disclosure, change or Substitutions should be included within the protection scope of the present disclosure. Therefore, the protection scope of the present disclosure should be subject to the protection scope defined by the claims.

Claims (14)

1. The crystal form III of the compound TAS-116 represented by the formula (I),

   

   Wherein, using Cu-Kα radiation, the X-ray powder diffraction pattern of the crystal form III has 2θ values of 6.39°±0.2°, 10.40°±0.2°, 12.94°±0.2° and 19.48°±0.2°. Characteristic peaks.
2. The crystal form III according to claim 1, wherein, using Cu-Kα radiation, the X-ray powder diffraction pattern of the crystal form III has at least 2θ values of 3.20°±0.2°, 6.39°±0.2°, 10.40 There is a characteristic peak at one of °±0.2°, 11.41°±0.2°, 12.94°±0.2° and 19.48°±0.2°.
3. The crystal form III according to claim 1 or 2, wherein, using Cu-Kα radiation, the X-ray powder diffraction pattern of the crystal form III is at least 3.20°±0.2°, 6.39°±0.2° in 2θ values , 10.40°±0.2°, 11.41°±0.2°, 12.94°±0.2°, 17.58°±0.2°, 18.26°±0.2°, 19.48°±0.2°, 21.71°±0.2°, 22.69°±0.2° and 23.65 One of the positions at °±0.2° has a characteristic peak.
4. The crystal form III according to any one of claims 1 to 3, wherein, using Cu-Kα radiation, the X-ray powder diffraction pattern of the crystal form III has a 2θ value of 3.20°±0.2°, 6.39° There are characteristic peaks at °±0.2°, 10.40°±0.2°, 11.41°±0.2°, 12.94°±0.2° and 19.48°±0.2°.
5. The crystal form III according to any one of claims 1 to 4, wherein, using Cu-Kα radiation, the X-ray powder diffraction pattern of the crystal form III has a 2θ value of 3.20°±0.2°, 6.39° °±0.2°, 10.40°±0.2°, 11.41°±0.2°, 12.94°±0.2°, 17.58°±0.2°, 18.26°±0.2°, 19.48°±0.2°, 21.71°±0.2°, 22.69°± There are characteristic peaks at 0.2° and 23.65°±0.2°.
6. The crystalline form III according to any one of claims 1 to 5, wherein, using Cu-Kα radiation, the X-ray powder diffraction pattern of the crystalline form III has characteristic peaks at the following diffraction angles, and its 2θ value and relative intensities are shown in the table below:

   
7. The crystalline form III of any one of claims 1 to 6, wherein the X-ray powder diffraction pattern of the crystalline form III is substantially as shown in FIG. 4 .
8. The crystal form III according to any one of claims 1 to 7, wherein the Fourier transform infrared spectrum of the crystal form III has a wavenumber of 1650 cm ^-1 ± 2 cm ^-1 , 1569 cm ^-1 ± 2 cm ^-1 , 1504cm ^-1 ±2cm ^-1 , 1424cm ^-1 ±2cm ^-1 , 1272cm ^-1 ±2cm ^-1 , 1030cm ^-1 ±2cm ^-1 , 823cm ^-1 ±2cm ^-1 , 812cm ^-1 ±2cm ^-1 , 748cm ^- There are characteristic peaks at ¹ ± 2 cm ^-1 , 711 cm ^-1 ± 2 cm ^-1 , 668 cm ^-1 ± 2 cm ^-1 and 653 cm ^-1 ± 2 cm ^-1 .
9. A method of preparing Form III according to any one of claims 1 to 8, comprising:

   The compound TAS-116 solid was formed into a suspension in solvent 1, stirred, the solid was separated, and dried to obtain the crystal form III;

   Preferably, the solvent 1 is selected from cyclic ethers; more preferably 1,4-dioxane;

   Preferably, the mass volume ratio of the compound TAS-116 solid to solvent 1 is 10 to 200:1;

   Preferably, the stirring time is 10 hours to 168 hours; more preferably 16 hours to 72 hours;

   Preferably, the stirring is carried out at room temperature;

   Preferably, after drying, the obtained solid is heated at a temperature of 60°C to 150°C, more preferably at a temperature of 80°C to 130°C, preferably, the heating time is 5 minutes to 16 hours.
10. A method of preparing Form III according to any one of claims 1 to 8, comprising:

    The compound TAS-116 solid is formed into a solution in solvent 2, the temperature is lowered, the solid is separated, and dried to obtain crystal form III;

    Preferably, the solvent 2 is selected from halogenated alkanes, alcohols, ketones, furans, cyclic ethers, nitriles or their mixed solvents; more preferably tetrahydrofuran and chloroform; preferably, two of the mixed solvents The volume ratio of the solvent is 1:4 to 4:1;

    Preferably, the mass volume ratio of the solute to the solvent 2 in the solution is 5 to 100:1;

    Preferably, the temperature for forming the solution is 60°C to 80°C;

    Preferably, the temperature is lowered to 10°C to 50°C, more preferably, the temperature is lowered to 10°C to 40°C;

    Preferably, the crystal form III seed crystal of TAS-116 is added during the cooling process;

    Preferably, the added amount of the seed crystal is 10% to 30% of TAS-116 solid;

    Preferably, the stirring time is 2 hours to 24 hours.
11. A pharmaceutical composition comprising the crystal form III according to any one of claims 1 to 8, or the crystal form III prepared by the method of claim 9 or 10, and at least one pharmaceutically acceptable vector.
12. A method for the treatment of cancer, comprising administering to an individual in need of the method a therapeutically marketable crystalline form III according to any one of claims 1 to 8, a crystalline form prepared by the method of claim 9 or 10 Type III or the pharmaceutical composition of claim 11.
13. The method of claim 12, wherein the cancer is selected from the group consisting of head and neck cancer, esophageal cancer, gastric cancer, colon cancer, rectal cancer, liver cancer, gallbladder-cholangiocarcinoma, biliary tract cancer, pancreatic cancer, lung cancer, breast cancer , ovarian cancer, cervical cancer, uterine cancer, kidney cancer, bladder cancer, prostate cancer, testicular tumors, bone-soft tissue sarcoma, leukemia, malignant lymphoma, multiple myeloma, skin cancer, brain tumor and mesothelioma.
14. The method of claim 12 or 13, further comprising administering to the individual at least one other drug for the treatment of cancer, preferably a monoclonal antibody drug, more preferably a PD-1 and PD-L1 inhibitor, even more preferably nivolumab and AB122.

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