AU2015392050B2

AU2015392050B2 - Fumarate of pyridylamine compound and crystals thereof

Info

Publication number: AU2015392050B2
Application number: AU2015392050A
Authority: AU
Inventors: Xinlu Li; Qingyi Meng; Xiquan Zhang; Rui Zhao
Original assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd
Current assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd
Priority date: 2015-04-23
Filing date: 2015-04-23
Publication date: 2020-01-23
Anticipated expiration: 2035-04-23
Also published as: RU2684278C1; CA2983475C; CA2983475A1; WO2016169030A1; AU2015392050A1

Abstract

Disclosed are a fumarate, an A-type crystal and B-type crystal of a pyridylamine compound having the structural formula (III), a preparation method therefor, medicine compositions and crystal compositions containing the compound and the crystals thereof, and uses thereof in preparation of medicines for preventing or treating tumors.

Description

FUMARATE OF PYRIDYLAMINE COMPOUND AND CRYSTALS THEREOF TECHNICAL FIELD

The present invention relates to a fumarate of a pyridylamine compound and a crystal

thereof, and belongs to the field of medical chemistry.

BACKGROUND

Protein Tyrosine Kinases (PTKs) play an extremely important role in the intracellular

signal transduction pathways. They involve in regulation, signal transmission and development of

normal cells, and are also closely related to proliferation, differentiation, migration and apoptosis of

tumor cells. Therefore, the inhibitory activities against protein tyrosine kinases have positive effects

on the inhibition and treatment of tumors. The protein tyrosine kinase family has a plurality of

subtypes, including epidermal growth factor receptor (EGFR) subtype, vascular endothelial growth

factor receptor (VEGFR) subtype, platelet-derived growth factor receptor (PDGFR) subtype,

anaplastic lymphoma kinase (ALK) and so on. The study has found that the abnormal activation and

expression of ALK kinases exist in various tumor cells such as non-small cell lung cancer, breast

cancer, glioblastomas and the like.

Crizotinib (XALKORITM) is an oral inhibitor of anaplastic lymphoma kinase (ALK)

developed by U.S. Pfizer Inc., which first launched in the United States in August 2011 (Nat. Rev.

Drug Discov. 10, 895-896, 2011). Clinically, the drug is mainly used to treat patients with anaplastic

lymphoma kinase (ALK)-positive locally advanced or metastatic non-small cell lung cancer

(NSCLC). The chemical structure of Crizotinib is shown as Formula (I): NH

N-N CI OIN

NH 2

F

Formula (I)

CN102850328A discloses a pyridylamine compound having the chemical structure as shown in Formula (II), which is an analogue of Crizotinib and has good inhibitory effects on ALK.

However, there are some problems associated with the compound of Formula (II), such as, ease of

absorbing moisture and the occurrence of degradation, stringent storage conditions and so on.

NH N-N CI ON

ciC HN-, F 0

Formula (II)

In addition to the therapeutic efficacy, the stability, hygroscopicity, bioavailability and

the like of a drug as a therapeutic agent in processing, manufacturing and storage are crucial to drug

research and development. Moreover, the chemical stability, solid-state stability and shelf life of an

active ingredient are very important factors from the viewpoint of obtaining a commercially viable

production method or from the viewpoint of producing a pharmaceutical composition comprising

an active compound. Therefore, it is very important for the production and storage of a drug to

provide a suitable form of the drug having desired properties.

SUMMARY OF THE INVENTION

The present invention provides a compound of Formula (III) having the following

structure, which is a fumarate of the compound of Formula (II): H

N-N

C1 OO Cl OHO OH NN HO"

C1 F 0

Formula (III)

The present invention further provides a method for preparing the compound of

Formula (III), comprising the following steps: dissolving the compound of Formula (II) in an

organic solvent, and adding a solution of fumaric acid in ethanol at 0°C to 80°C to carry out a

reaction to obtain the compound of Formula (III).

In some embodiments of the present invention, the solution of fumaric acid in ethanol

is added under stirring, and the reaction is carried out for 0.5 hour to 2 hours.

In some embodiments of the present invention, after solids are precipitated out of the

reaction solution, the solids are filtered and dried, preferably dried under vacuum.

In some embodiments of the present invention, the organic solvent is one or more

selected from the group consisting of methanol, ethanol, dichloromethane and acetone.

In some embodiments of the present invention, the molar ratio of the added fumaric

acid to the compound of Formula (II) is 1.2-1.5:1.

In some embodiments of the present invention, the concentration of the added solution

of fumaric acid in ethanol is 1.0 mol/L to 2.0 mol/L, preferably 1.0 mol/L to 1.5 mol/L.

In some embodiments of the present invention, the reaction is preferably carried out at

0°C to 50°C; in some other embodiments, the reaction is more preferably carried out at 20°C to

50°C.

In certain particular embodiments, the compound of Formula (III) may be prepared

according to the following steps: dissolving the compound of Formula (II) in ethanol, adding a

solution of fumaric acid in ethanol at 20 0 C to 500 C under stirring, reacting for 0.5 hour to 2 hours,

precipitating solids out of the reaction solution, filtering the solids, and drying the solids under

vacuum to obtain the compound of Formula (III).

The fumaric acid can be replaced with different inorganic or organic acids, and various

acid addition salts of the compound of Formula (II) can be prepared by using methods similar to

those described above.

The present invention further provides a pharmaceutical composition comprising a

therapeutically effective amount of the compound of Formula (III) and a pharmaceutically

acceptable carrier. The pharmaceutically acceptable carrier may be solid or liquid. The solid carrier

may comprise one or more selected from the group of flavoring agents, lubricants, solubilizers,

suspending agents, fillers, binders, tablet disintegrating agents or encapsulating materials. Suitable

solid carriers include, for example, magnesium stearate, talc, sucrose, lactose, dextrin, starch,

gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidone. The liquid carriers are used to prepare compositions such as solutions, suspensions, emulsions, syrups and the like. Suitable liquid carriers for oral and parenteral administration include water, alcohols, oil and the like. The present invention further provides use of the compound of Formula (III) in the preparation of a medicament for the prophylaxis or treatment of tumors. The compound of Formula

(III) of the present invention may be used alone or in combination with other drugs for preparing

anti-tumor medicaments. The tumors may be lung cancers, preferably ALK-positive primary or

metastatic non-small cell lung cancers.

The present invention also provides a crystalline Form A of the compound of Formula

(III), NH N-N

0 CI • HO x OH N0 o

F 0

Formula (III)

characterized in that the X-ray powder diffraction spectrum thereof has diffraction peaks expressed

by 20 values at about 6.3, 11.7, 12.5, 14.1, 22.6 and 23.3; typically has diffraction peaks

expressed by 20 values at about 6.3, 11.7°, 12.5°, 14.1°, 19.7°, 21.2°, 22.6°, 23.3°, 23.8° and 25.5°;

more typically has diffraction peaks expressed by 20 values at about 6.3°, 11.7°, 12.5°, 14.1°, 15.0°,

15.90, 17.00, 19.7, 20.6, 21.2, 21.6, 22.6, 23.3, 23.80 and 25.5°; and more typically has

diffraction peaks expressed by 20 values at about 6.3°, 9.9, 11.7°, 12.5°, 14.1°, 15.0°, 15.9°, 17.0°,

19.7, 20.6, 21.2, 21.6, 22.60, 23.30, 23.80, 24.60, 25.10, 25.50, 27.10 and 28.7.

A typical but non-limited example of the crystalline Form A of the compound of

Formula (III) provided in the present invention has a differential scanning calorimetry (DSC)

thermogram with an absorption peak at about 227.5°C.

A typical but non-limited example of the crystalline Form A of the compound of

Formula (III) provided in the present invention has an infrared (IR) spectrum as shown in FIG. 5.

In another aspect, the present invention provides a crystalline composition, wherein the

above crystalline Form A of the compound of Formula (III) accounts for 50% or more, preferably

80% or more, more preferably 90% or more, and most preferably 95% or more, by weight of the

crystalline composition.

In another aspect, the present invention provides a pharmaceutical composition

comprising a therapeutically effective amount of the above crystalline Form A of the compound of

Formula (III) or the above crystalline composition.

In another aspect, the present invention provides use of the above crystalline Form A of

the compound of Formula (III), the above crystalline composition or the above pharmaceutical

composition in the preparation of a medicament for the prophylaxis or treatment of tumors,

preferably in the preparation of a medicament for the prophylaxis or treatment of lung cancers, and

more preferably in the preparation of a medicament for the prophylaxis or treatment of

ALK-positive primary or metastatic non-small cell lung cancers.

In another aspect, the present invention provides a method for preparing the above

crystalline Form A of the compound of Formula (III) or the above crystalline composition,

comprising:

(a) dissolving the compound of Formula (III) in an organic solvent, and heating the

resulting solution under stirring;

(b) adding a second solvent; and

(c) cooling the resulting mixture to precipitate crystals.

In the above step (a), the organic solvent is a lower alcohol, preferably a C1 -C 4 alkyl

alcohol, and more preferably methanol; in the step (a), the resulting solution may be heated to 40°C

to 65°C, preferably 50°C to 60°C; in the step (a), the ratio of the compound of Formula (III) to the

organic solvent is preferably 1g/50 ml to 1 g/10 ml, more preferably 1 g/15 ml to 1 g/10 ml; in the

step (a), the rate of the stirring is preferably 300 r/min to 500 r/min; in the step (b), preferably, the

second solvent is acetone, tetrahydrofuran, dioxane or water; the volume ratio of the second solvent

in the step (b) to the organic solvent in the step (a) may be 1-3:1, preferably 2:1; in the step (c), the

resulting mixture may be cooled to -15°C to 0°C, preferably cooled to 0°C.

In some embodiments of the present invention, the above method for preparing the

crystalline Form A of the compound of Formula (III)may further comprise:

(d) filtering and drying.

In the step (d), the drying may be performed at 45°C under vacuum or by blowing air at

45°C under atmospheric pressure.

The present invention also provides a crystalline Form B of the compound of Formula

(III), NH N-N C O HO OH

F 0

Formula (III)

characterized in that the X-ray powder diffraction spectrum of the crystalline Form B has diffraction

peaks expressed by 20 values at about 23.0°, 24.9°, 25.9°, 27.0°, 28.9°, 29.5, 38.1° and 38.8°;

typically has diffraction peaks expressed by 20 values at about 18.7°, 23.0°, 24.9°, 25.9°, 27.0°,

28.0, 28.9, 29.5°, 36.0°, 38.1° and 38.7°.

A typical but non-limited example of the crystalline Form B of the compound of

thermogram with an absorption peak at about 230.6°C.

A typical but non-limited example of the crystalline Form B of the compound of

Formula (III) provided in the present invention has an infrared (IR) spectrum as shown in FIG. 6.

above crystalline Form B of the compound of Formula (III) accounts for 50% or more, preferably

crystalline composition.

In another aspect, the present invention provides a pharmaceutical composition

comprising a therapeutically effective amount of the above crystalline Form B of the compound of

Formula (III) or the above crystalline composition.

In another aspect, the present invention provides use of the above crystalline Form B of

ALK-positive primary or metastatic non-small cell lung cancers.

crystalline Form B of the compound of Formula (III) or the above crystalline composition,

comprising:

(a) dissolving the compound of Formula (III) in anhydrous methanol with heating and

stirring; and

(b) step-cooling the resulting solution to precipitate crystals.

In the above step (a), the heating may be carried out at 40°C to 70°C, and preferably

the heating is carried out under reflux; the rate of the stirring is preferably 300 r/min to 500 r/min;

the ratio of the compound of Formula (III) to the anhydrous methanol is preferably 1 g/50 ml to 1

g/10 ml, more preferably 1 g/15 ml to 1 g/10 ml. In the above step (b), the step-cooling may be so

that the resulting solution is cooled to 15°C to 25°C, and further cooled to -5°C to -20°C; preferably

cooled to room temperature, and further cooled to -18 °C.

crystalline Form B of the compound of Formula (III) may further comprise:

(c) filtering and drying.

In the step (c), the drying may be performed at 45°C under vacuum or by blowing air at

45°C under atmospheric pressure.

In the present invention, the X-ray powder diffraction spectrum of a sample is

measured under the following conditions:

Instrument: Bruker D2 X-ray diffractometer; Test Conditions: 30 kv 10 mA; Slit:

0.6mm/3mm/0.8mm; Target Type: Cu; Angle Range: 5°C to 400; Step Size: 0.1 s/0.02°.

In the present invention, IHNMR is measured under the following conditions:

Test Organization: Analysis and Test Center of China Pharmaceutical University;

Instrument: BRUKER AV-500 type nuclear magnetic resonance spectrometer; Solvent: DMSO-d6;

Internal Standard Substance: TMS; Temperature: 303 K.

In the present invention, the DSC spectrum is measured under the following conditions:

Instrument: Mettler type 1 differential thermal analyzer; Temperature Range: 30°C to

270°C; Heating Rate: 10 °C/min.

In the present invention, the IR spectrum is measured under the following conditions:

Instrument: Perkin Elmer spetrum type 100 infrared spectrometer; Instrument

Calibration: the wave number of the instrument is calibrated with the absorption peak of the infrared spectrum of polystyrene film; Method: KBr pellet pressing method.

In the present invention, unless specifically stated, otherwise, ethanol used herein is

anhydrous ethanol.

It should be noted that, in an X-ray powder diffraction (XRD) spectrum, a diffraction

pattern of a crystalline compound is usually characteristic for a specific crystalline form. Relative

intensities of the bands (especially at the low angles) can vary depending upon preferential

orientation effects resulting from the differences of crystals' conditions, particle sizes, and other

measuring conditions. Therefore, the relative intensities of diffraction peaks are not characteristic

for a specific crystalline form. It is the relative positions of peaks rather than relative intensities

thereof that should be paid more attention when judging whether a crystalline form is the same as a

known crystalline form. In additional, as for any given crystalline form, there may be a slight error

in the position of peaks, which is also well known in the field of crystallography. For example, the

position of a peak may shift due to the change of a temperature, the movement of a sample or the

calibration of an instrument and so on when analyzing the sample, and the measurement error of 20

value is sometimes about 0.2. Accordingly, this error should be taken into consideration when

identifying a crystal structure. Usually, the position of a peak is expressed in terms of 20 angle or

lattice spacing d in an XRD pattern and the simple conversion relationship therebetween is d =

X/2sin, wherein d represents the lattice spacing, k represents the wavelength of incident X-ray, and

0 represents the diffraction angle. For the same crystalline form of the same compound, the position

of peaks in an XRD spectrum thereof has similarity on the whole, and the error of relative

intensities may be larger. In addition, it is necessary to point out that due to some factors such as

reduced contents, parts of diffraction lines may be absent in the identification of a mixture. At this

time, even a band may be characteristic for the given crystalline form without depending upon all

the bands of a high purity sample.

DSC is used to measure a thermal transition temperature when absorbing or releasing

heat due to the change of a crystal structure or the melting of a crystal. In a continuous analysis of

the same crystalline form of the same compound, the error of a thermal transition temperature and a

melting point is typically within a range of about 5°C, generally within a range of about 3°C. A

compound with a given DSC peak or melting point means that the DSC peak or melting point may

be varied within a range of 5°C. DSC provides an auxiliary method to distinguish different

crystalline forms. Different crystalline forms can be identified by their characteristically different transition temperatures. It is necessary to point out that the DSC peak or melting point of a mixture may vary over a wider range. Furthermore, because of the decomposition in the melting process of a substance, the melting temperature is related to a heating rate.

In the present invention, the compound of Formula (II) can be prepared with reference

to the method disclosed in CN201210128875.0.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an X-ray powder diffraction pattern of the crystalline Form A of the

compound of Formula (III) prepared in Example 3.

FIG. 2 is an X-ray powder diffraction pattern of the crystalline Form B of the

compound of Formula (III) prepared in Example 5.

FIG. 3 is a differential scanning calorimetry (DSC) thermogram of the crystalline Form

A of the compound of Formula (III) prepared in Example 3.

FIG. 4 is a differential scanning calorimetry (DSC) thermogram of the crystalline Form

B of the compound of Formula (III) prepared in Example 5.

FIG. 5 is an infrared (IR) spectrum of the crystalline Form A of the compound of

Formula (III) prepared in Example 3.

FIG. 6 is an infrared (IR) spectrum of the crystalline Form B of the compound of

Formula (III) prepared in Example 5.

EMBODIMENTS

The present invention will be described in further detail with reference to the following

examples, but the present invention is not limited to the following examples.

Example 1 Preparation of the compound of Formula (II)

A. Preparation of N-acetyl-5-bromo-3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]-2

pyridylamine Br

CI HN

F C HN-CH3 F

500 mg of 5-bromo-3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethyoxy]-2-pyridylamine

was dissolved in 15 ml of dichloromethane. The resulting mixture was cooled to 0C. 1 ml of

triethylamine was added and the resulting mixture was continuously stirred for 5 minutes. And then,

1.1 equivalents of acetyl chloride was added dropwise. The resulting mixture was warmed to room

temperature and reacted for 5 hours. The reaction was quenched by adding water. The resulting

mixture was extracted with dichloromethane, dried with anhydrous sodium sulfate, filtered and

concentrated. The resulting crude product was purified by column chromatography (ethyl acetate:

petroleum ether = 1: 4) to obtain 400 mg of a yellow-white solid (yield 72%).

5-Bromo-3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethyoxyl]-2-pyridylamine can be

prepared according to the methods disclosed in the literatures, for example, Organic Process

Research & Development, 2011, 15(5):1018-1026 or W02007066187 and so on.

B. Preparation of N-acetyl-3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]-5-[1

(4-N-Boc-piperidyl)-1H-pyrazol-4-yl]-2-pyridylamine Boc

N-N CI O N

C1 N F

300 mg of N-acetyl-5-bromo-3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]-2

pyridylamine and 230 mg of 1-(4-N-Boc-piperidyl)-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)

-1H-pyrazole were dissolved in 5 ml of DMF, and the resulting mixture was added into 1 ml of an

aqueous solution containing 300 mg of cesium carbonate. The air was replaced with nitrogen for

three times, and 20 mg of Pd(PPh3) 2Cl2 was added, then the air was replaced with nitrogen for three

times again. The resulting reaction mixture was heated to 75°C and stirred for 12 hours. After the

reaction was completed, the reaction mixture was cooled to room temperature and then diluted by

adding 20 ml of ethyl acetate, filtered with celite and washed with ethyl acetate. The combined

ethyl acetate layer was dried over anhydrous sodium sulfate and then concentrated. The resulting

crude product was purified by column chromatography (ethyl acetate: petroleum ether = 1: 1) to

obtain 330 mg of a white foam solid (yield 78%).

C. Preparation of N-acetyl-3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]-5-[1-

(4-piperidyl)-1H-pyrazol-4-yl]-2-pyidylamine H

N-N I, CI NH F

100 mg of the obtained N-acetyl-3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]-5

[1-(4-N-Boc-piperidyl)-1H-pyrazol-4-yl]-2-pyridylamine was dissolved in a small amount of

dichloromethane. Under stirring at 0C, 2 ml of a solution of 4N HCl in dioxane was added, the

resulting mixture was stirred for 20 minutes, and then the solvent was removed under reduced

pressure. 10 ml of water was added, and the resulting mixture was adjusted with sodium

bicarbonate solid to pH=10, extracted by using dichloromethane and then dried, concentrated and

purified by column chromatography to obtain 71 mg of a white solid (yield 85%). MS: m/e 492 (M

+1).

Example 2 Preparation of the compound of Formula (III)

0.01mol of the compound of Formula (II) was dissolved in ethanol, and 8.6 ml of a

1.4mol/1 pre-prepared solution of fumaric acid in ethanol was added at 20°C under stirring. After

the reaction was performed for 1 hour, solids were precipitated out of the reaction solution, filtered,

and dried at 45°C under vacuum to obtain an off-white solid (yield 85.5%). 1HNMR (DMSO-d6) 6 (ppm): 1.79(3H, -CH3), 2.10(3H, -CH 3), 2.19(4H, -CH 2 -),

3.04(2H, -CH 2 -), 3.38(2H, -CH 2-), 4.50(1H, -CH-), 6.16(1H, -CH-), 6.52(2H, -CH), 7.38(1H, ArH),

7.44(1H, ArH), 7.56(1H, ArH), 7.83(1H, =CH-N), 8.23(1H, -CH=N), 8.24(1H, -NHCO-), 9.39(1H,

ArH), 9-12(3H, 2*-COOH, -NH-).

Example 3 Preparation of the crystalline Form A of the compound of Formula (III)

2.0 g of the compound of the Formula (III) was dissolved in 20 ml of methanol. The

resulting solution was heated to 50°C under stirring, and then 40 ml of acetone was added. The

resulting mixture was cooled to 0°C, and kept for 48 h, and crystals were precipitated out, filtered

and dried at 45°C under vacuum to obtain a crystalline Form A (yield 87.7%). The X-ray powder

diffraction pattern of the obtained crystal is shown in FIG. 1. The differential scanning calorimetry

(DSC) thermogram of the obtained crystal is shown in FIG. 3. The infrared (IR) spectrum of the

obtained crystal is shown in FIG. 5.

Example 4 Preparation of the crystalline Form A of the compound of Formula (III)

2.0 g of the compound of Formula (III) was dissolved in 20 ml of methanol. The

resulting solution was heated to 50°C under stirring, and then 40 ml of tetrahydrofuran was added.

The resulting mixture was cooled to 0°C, and kept for 48 h, and crystals were precipitated out,

filtered and dried by blowing air at 45°C under atmospheric pressure. X-ray powder diffraction

analysis showed that the obtained crystal is a crystalline Form A (yield 86.0%).

Example 5 Preparation of the crystalline Form B of the compound of Formula (III)

5.0 g of the compound of Formula (III) was added in 50 ml of anhydrous methanol.

The resulting mixture was heated to reflux under stirring so that the compound of Formula (III) was

dissolved, and the resulting solution was gradually cooled to room temperature and then further

cooled to -18°C. The mixture was kept standing at the low temperature for 48 hours, and crystals

were precipitated out, filtered, dried by blowing air at 45°C under atmospheric pressure for 4 hours

to obtain a crystalline Form B (yield 86.4%). The X-ray powder diffraction pattern of the obtained

crystal is shown in FIG. 2. The differential scanning calorimetry (DSC) thermogram of the obtained

crystal is shown in FIG. 4. The infrared (IR) spectrum of the obtained crystal is shown in FIG. 6.

Reference Example 1 Preparation of other acid addition salts of the compound of Formula

(II) A series of acid addition salts of the compound of Formula (II) were obtained by using

the method of Example 2, except for replacing fumaric acid with different organic or inorganic

acids.

Table 1 Other acid addition salts of the compound of Formula (II) Acid addition salts of the Melting points of the compound of Formula (II) products (m.p.: °C) Hydrochloride off-white solid 138.3-139.2 p-Toluenesulfonate off-white solid 136.7-138.6 Mesylate off-white solid 143.0-144.7

Maleate off-white solid 177.1-178.8 Malate off-white solid 124.7-127.6 Succinate off-white solid 77.9-79.2

Example 6 Hygroscopicity Test

The compound of Formula (II) and various salts of the compound of Formula (II)

prepared in Example 2 and Reference Example 1 were tested according to "Guiding Principles for

Drug Hygroscopicity Test" described in the Chinese Pharmacopoeia, 2010 edition, Part II,

Appendix XIX J. The increased weights by hygroscopy of the samples were calculated respectively,

and the test results are shown in Table 2.

Table 2 Hygroscopicity Test Results Sample Names Increased Weights by Hygroscopy(%) Compound of Formula (II) 61.75 Fumarate 0.80 Hydrochloride 256.39 p-Toluenesulfonate 164.27 Mesylate 232.53 Maleate 1.15 Malate 7.83 Succinate 3.60

Example 7 Pharmacokinetic test of the acid addition salts of the compound of Formula (1)

Twenty-seven healthy SD male rats, weighting 200g-220g, were fed with standard

formula rat pellet feed at fixed times daily. The rats were fasted for 12 h before the experiment, and

fed again 4 hours after the administration of the drug. The rats can drink water freely before and

after the experiment as well as during the course of the experiment. The rats were randomly divided

into 9 groups. A single dose of Crizotinib was intragastrically administered to the first group; a

single dose of the compound of Formula (II) was intragastrically administered to the second group;

and a single dose of the compound of Formula (III),and the hydrochloride, p-toluenesulfonate,

mesylate, maleate, malate, and succinate of the compound of Formula (II)were intragastrically

administered to the third group to the ninth group, respectively. All the doses administered in the 9

groups of the rats are 0.11 mmol/kg, and 0.2 mL to 0.3 mL blood was taken from ocular fundus venous plexus before the administration of the drug (0h) and 0.5 h, 1h, 2 h, 4 h, 6 h, 8 h, 10 h and 24 h after the administration of the drug. Heparin was used for the anti-coagulation of the blood, and the blood was centrifuged to separate blood plasma. 0.1 mL of the blood plasma was accurately measured and added into an EP tube, and then 1.2 ml of ethyl acetate was added. The resulting mixture was uniformly mixed at a high speed for 5 min by a vortex mixer, and centrifuged for 5 min (8000 r-min-). The supernatant liquid was collected, and the solvent was blown-dry at 30°C with nitrogen on a nitrogen blowing instrument. The residue was dissolved with 100 L of a mobile phase, uniformly mixed at a high speed for 1 min by a vortex mixer, and centrifuged for 5 min (14000 r-min-'). 80iL of the supernatant liquid was transferred to a sample vial, and 10 L thereof was injected for HPLC detection, and the HPLC chromatogram was recorded. The results are shown in Table 3: Table 3 Oral bioavailability results of the compounds in rats Oral Bioavailabilities Compounds (AUC, ug/mL*h) Crizotinib 15.474 Compound of Formula (II) 17.056 Fumarate 25.654 Hydrochloride 19.043 p-Toluenesulfonate 13.182 Mesylate 12.866 Maleate 14.165 Malate 17.393 Succinate 10.453

The HPLC detection conditions are as follows: Liquid Chromatograph: Shimadzu Ultra-fast High Performance Liquid Chromatography Prominence UFLC XR Analysis Column: Shim-pack XR-ODS 11 (2.0*75mm 2.2pm) Mobile Phase: 0.1% formic acid solution containing 5 mM ammonium formate/acetonitrile = 80/20 (V/V) Flow Rate: 0.25 mL/min, Column Temperature: 40°C Sample Volume: 10 pL, Analysis Time: 10.5 min Wavelength Range of PDA: 260 nm to 275 nm, Temperature of Detection Cell: 40°C

Example 8 Stability Test

According to the test method of the influence factors for active pharmaceutical

ingredients described in the Chinese Pharmacopoeia, 2010 edition, Part II, Appendix XIX C, the

crystalline Form A of the compound of Formula (III) prepared in Example 3 and the crystalline

Form B of the compound of Formula (III) prepared in Example 5 were subjected to a

high-temperature test (40°C ±2C, a relative humidity of 75% ±5%) and strong light irradiation

test (45001x 5001x), respectively. The tests last for 10 days. Samples were taken on days 0 and 10

to measure a total amount of impurities so as to determine their stabilities. The test results are

shown in Table 4.

Table 4 Stability Test Results Test Items Crystalline Form A Crystalline Form B

Day 0 0.73 0.65 Total amount of Light Irradiation for impurities (%) 0.89 1.62 10 days At 40 °C for 10 days 0.85 2.56

By way of clarification and for avoidance of doubt, as used herein and except where the

context requires otherwise, the term "comprise" and variations of the term, such as "comprising",

"comprises" and "comprised", are not intended to exclude further additions, components, integers or

steps.

Reference to any prior art in the specification is not an acknowledgement or suggestion

that this prior art forms part of the common general knowledge in any jurisdiction or that this prior

art could reasonably be expected to be combined with any other piece of prior art by a skilled

person in the art.

Claims

What is claimed is 1. A compound of Formula (III) having the following structure:

NH

N-N

0 CI O

C N HO F O

Formula (III)
2. A method for preparing a compound of Formula (III), characterized in that the

method comprises the following steps:

dissolving a compound of Formula (II) in an organic solvent, and adding a solution of

fumaric acid in ethanol to carry out a reaction at 0°C to 80°C, to obtain the compound of Formula

(III), wherein the reaction is carried out for 0.5 hour to 2 hours, NH NH

N-N N-N

Cl CI 0~~ 0 N H 0

CI HIN,,r CI HN F 0 F 0

Formula (II) Formula (III)
3. The method according to Claim 2, characterized in that the organic solvent is one or

more selected from the group consisting of methanol, ethanol, dichloromethane and acetone.
4. The method according to Claim 2 or 3, characterized in that the molar ratio of the

added fumaric acid to the compound of Formula (II) is 1.2-1.5:1.
5. The method according to any one of Claims 2 to 4, characterized in that the concentration of the solution of fumaric acid in ethanol is 1.0 mol/L to 2. 0 mol/L.
6. A crystalline Form A of a compound of Formula (III) NH

N-N

0 CI H OH

HN C1 F 0

Formula (III)

characterized in that the X-ray powder diffraction spectrum thereof has diffraction

peaks expressed by 20 values at about 6.3, 11.7, 12.5, 14.10, 22.6 and 23.3.
7. The crystalline Form A of the compound of Formula (III) according to Claim 6,

characterized in that the X-ray powder diffraction spectrum thereof has diffraction peaks expressed

by 20 values at about 6.3, 11.7, 12.5, 14.10, 19.7, 21.2, 22.6, 23.3, 23.8 and 25.5.
8. The crystalline Form A of the compound of Formula (III) according to Claim 6,

characterized in that the X-ray powder diffraction spectrum thereof has diffraction peaks expressed

by 20 values at about 6.3, 11.7, 12.5, 14.10, 15.0, 15.9, 17.0, 19.7, 20.6, 21.2, 21.6, 22.6,

23.3, 23.80 and 25.5.
9. The crystalline Form A of the compound of Formula (III) according to Claim 6,

characterized in that the X-ray powder diffraction spectrum thereof has diffraction peaks expressed

by 20 values at about 6.30, 9.90, 11.7, 12.5, 14.1, 15.00, 15.90, 17.00, 19.7, 20.6, 21.2, 21.6,

22.6, 23.3, 23.8, 24.6, 25.10, 25.50, 27.10 and 28.7.
10. The crystalline Form A of the compound of Formula (III) according to Claim 6,

characterized in that the differential scanning calorimetry thermogram thereof has an absorption peak at about 227.5°C.
11. A method for preparing the crystalline Form A of the compound of Formula (III)

according to any one of Claims 6 to 10, comprising:

(a) dissolving the compound of Formula (III) in an organic solvent, and heating the

resulting solution under stirring;

(b) adding a second solvent; and

(c) cooling the resulting mixture to precipitate crystals.
12. The method according to Claim 11, characterized in that, in the step (a), the organic

solvent is a C1-C4 alkyl alcohol; and/or the resulting solution is heated to 40°C to 65°C; and/or the

ratio of the compound of Formula (III) to the organic solvent is lg/50ml to lg/1Oml; and/or the rate

of the stirring is 300 r/min to 500 r/min.
13. The method according to Claim 11 or 12, characterized in that, in the step (b), the

second solvent is acetone, tetrahydrofuran, dioxane or water; and/or the volume ratio of the second

solvent in the step (b) to the organic solvent in the step (a) is 1-3:1; and/or in the step (c), the

resulting mixture is cooled to -15°C to 0°C.
14. A crystalline Form B of a compound of Formula (III)

NH

I7N

0 CI H x)OH N

C0 F 0

Formula (III)

characterized in that the X-ray powder diffraction spectrum of the crystalline Form B

has diffraction peaks expressed by 20 values at about 23.0°, 24.9, 25.9, 27.0, 28.9, 29.5, 38.1

and 38.80.
15. The crystalline Form B of the compound of Formula (III) according to Claim 14,

characterized in that the X-ray powder diffraction spectrum of the crystalline Form B has diffraction

peaks expressed by 20 values at about 18.7, 23.0, 24.9, 25.9, 27.0, 28.0, 28.9, 29.5, 36.0,

38.10 and 38.7.
16. A method for preparing the crystalline Form B of the compound of Formula (III)

according to Claim 14 or 15, comprising:

(a) dissolving the compound of Formula (III) in anhydrous methanol with heating and

stirring; and

(b) step-cooling the resulting solution to precipitate crystals.
17. The method according to Claim 16, characterized in that, in the step (a), the heating

is carried out at 40°C to 70°C; and/or the rate of the stirring is 300 r/min to 500 r/min; and/or the

ratio of the compound of Formula (III) to the anhydrous methanol is 1 g/15 ml to 1 g/10 ml; and

in the step (b), the step-cooling is so that the resulting solution is cooled to 15°C to 25°C, and

further cooled to -5°C to -20°C.
18. A crystalline composition, wherein the crystalline Form A according to any one of

Claims 6 to 10, or the crystalline Form B according to any one of Claims 14 to 15 accounts for 80%

or more, by weight of the crystalline composition.
19. A pharmaceutical composition, comprising a therapeutically effective amount of the

compound of Formula (III) according to Claim 1, the crystalline Form A according to any one of

Claims 6 to 10, the crystalline Form B according to any one of Claims 14 to 15, or the crystalline

composition according to Claim 18.
20. Use of the compound of Formula (III) according to Claim 1, the crystalline Form A

according to any one of Claims 6 to 10, the crystalline Form B according to any one of Claims 14 to

15, the crystalline composition according to Claim 18 or the pharmaceutical composition according

to Claim 19 in the preparation of a medicament for the prophylaxis or treatment of tumors; wherein the tumors are ALK-positive primary or metastatic non-small cell lung cancers.
21. A method of preventing or treating tumors comprising administering to a subject a

therapeutically effective amount of the compound of Formula (III) according to Claim 1, the

crystalline Form A according to any one of Claims 6 to 10, the crystalline Form B according to any

one of Claims 14 to 15, the crystalline composition according to Claim 18 or the pharmaceutical

composition according to Claim 19; wherein the tumors are ALK-positive primary or metastatic

non-small cell lung cancers.