JP2016034901A - Novel crystal form of 3,4-difluoro-2-(2-fluoro-4-iodo-phenyl amino)-n-(2-hydroxy-ethoxy)-5-(3-oxo-[1,2]oxazinan-2-ylmethyl)-benzamide - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an active pharmaceutical ingredient that is useful as MEK inhibitors and is stable physicochemically.SOLUTION: The present invention provides an educt I-type crystal of 3,4-difluoro-2-(2-fluoro-4-iodo-phenyl amino)-n-(2-hydroxy-ethoxy)-5-(3-oxo-[1,2]oxazinan-2-ylmethyl)-benzamide, having peaks with diffraction angles expressed by 2θ of 14.9, 16.9 and 23.1° (±0.2°) in a powder X-ray diffraction pattern measured using Cu Kα(1.54060Å) as an X-ray source.SELECTED DRAWING: None

Description

  The present invention relates to 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1,2] oxazinane-2 -Ilmethyl) -benzamide (hereinafter referred to as "Compound A") relating to a crystalline form referred to as Form I.

MEK has attracted attention as a target for therapeutic agents in the field of treatment of diseases caused by abnormal cell proliferation, and many compounds for MEK inhibition have been reported so far (Patent Documents 1 to 3). One of the MEK inhibitors, 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1,2 ] Oxazinan-2-ylmethyl) -benzamide (Compound A) is useful for the treatment of proliferative diseases such as cancer, psoriasis, restenosis, autoimmune diseases and atherosclerosis, as well as heart failure sequelae, xenografts It is known to be useful for the treatment of rejection, osteoarthritis, rheumatoid arthritis, asthma, cystic fibrosis, hepatoma, cardiac hypertrophy, Alzheimer's disease, diabetes, septic shock, HIV infection ( Patent Document 4).
Patent Document 4 discloses 5-substituted-2-phenylamino-benzamides having a MEK inhibitory effect, and also describes a synthesis method of Compound A. There is no disclosure regarding properties.

International Publication No. 98/37881 International Publication No.99 / 01421 International Publication No. 2000/35435 International Publication No. 2006/011466

  The inventors of the present invention have conducted extensive research for the purpose of providing a medicinal compound having a MEK inhibitory activity effect. As a result of further investigation, it was found that Compound A exhibits an excellent MEK inhibitory effect. The crystal of A has many polymorphs, and among these, the crystal form designated as Form I was found to be excellent in physicochemical stability and useful as a MEK inhibitor. That is, the present invention provides a MEK inhibitor excellent in physicochemical stability.

Specifically, the present invention includes the following.
(1) 3,4-Difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1,2] oxazinane-2- (Ilmethyl) -benzamide crystals.

(2) 3,4-Difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1,2] oxazinane-2- 2. The crystal according to 1 above, wherein ylmethyl) -benzamide is in a free form.

(3) The crystal as described in 2 above, which contains peaks having diffraction angles represented by 2θ of 14.9, 16.9 and 23.1 ° (± 0.2 °) in a powder X-ray diffraction pattern.

(4) The crystal according to 3 above, wherein the powder X-ray diffraction pattern further comprises peaks having diffraction angles represented by 2θ of 12.5, 18.0 and 22.7 ° (± 0.2 °).

(5) The crystal according to 4 above, wherein the powder X-ray diffraction pattern further includes peaks whose diffraction angles represented by 2θ are 13.0, 19.9, and 22.4 ° (± 0.2 °).

(6) The crystal as described in 5 above, wherein the peak temperature of the endothermic peak in DSC measurement (temperature increase rate: 10 ° C / min) is from 137 ° C to 147 ° C.

(7) The crystal according to 1 above, wherein the powder X-ray diffraction pattern includes peaks having diffraction angles represented by 2θ of 5.4, 6.2, 11.1, 15.4, 17.4, and 21.1 ° (± 0.2 °).

(8) The crystal according to 1 above, wherein the powder X-ray diffraction pattern includes peaks having diffraction angles represented by 2θ of 5.8, 10.0, 10.9, 11.5, 11.9, and 20.5 ° (± 0.2 °).

(9) The crystal according to 1 above, wherein the powder X-ray diffraction pattern includes peaks having diffraction angles represented by 2θ of 7.9, 11.7, 13.6, 17.9, 20.9, and 23.8 ° (± 0.2 °).

(10) The crystal according to 1 above, wherein the powder X-ray diffraction pattern includes peaks having diffraction angles represented by 2θ of 8.3, 10.7, 12.4, 14.2, 16.5, and 21.8 ° (± 0.2 °).

(11) A pharmaceutical composition comprising the crystal according to any one of 1 to 10 above.

By using form I crystals with suitable properties as pharmaceuticals as active pharmaceutical ingredients, it is possible to provide stable pharmaceuticals without stabilization by special pharmaceutical treatments, special packaging, special storage conditions, etc.

FIG. 1 shows 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [ 1 is an example of a powder X-ray diffraction pattern (X-ray source: Cu Kα) of a free form I crystal of [1,2] oxazinan-2-ylmethyl) -benzamide. FIG. 2 shows 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [ 1 is an example of a powder X-ray diffraction pattern (X-ray source: Cu Kα) of a free form II crystal of [1,2] oxazinan-2-ylmethyl) -benzamide. FIG. 3 shows 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [ 1 is an example of a powder X-ray diffraction pattern (X-ray source: Cu Kα) of a free form III crystal of 1,2] oxazinan-2-ylmethyl) -benzamide. FIG. 4 shows 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [ 1 is an example of a powder X-ray diffraction pattern (X-ray source: Cu Kα) of a free form IV crystal of 1,2] oxazinan-2-ylmethyl) -benzamide. FIG. 5 shows 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [ 1 is an example of a powder X-ray diffraction pattern (X-ray source: Cu Kα) of sodium salt tetrahydrate crystal of [1,2] oxazinan-2-ylmethyl) -benzamide. FIG. 6 shows 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [ 1 is an example of a DSC chart (heating rate: 10 ° C./min) of a free form I crystal of [1,2] oxazinan-2-ylmethyl) -benzamide. FIG. 7 shows 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [ 1 is an example of a DSC chart (temperature increase rate: 10 ° C./min) of a free form II crystal of [1,2] oxazinan-2-ylmethyl) -benzamide. FIG. 8 shows 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [ 1 is an example of a DSC chart (heating rate: 10 ° C./min) of a free form III crystal of [1,2] oxazinan-2-ylmethyl) -benzamide. FIG. 9 shows 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [ 1 is an example of a DSC chart (temperature increase rate: 10 ° C./min) of a free form IV crystal of [1,2] oxazinan-2-ylmethyl) -benzamide. FIG. 10 shows 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [ 1 is an example of a DSC chart (temperature increase rate: 10 ° C./min) of sodium salt tetrahydrate crystal of [1,2] oxazinan-2-ylmethyl) -benzamide.

  In the present invention, “3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1,2] oxazinane- “2-ylmethyl) -benzamide” (compound A) means a compound represented by the following chemical structural formula (I).

  Compound A can be produced based on the production method described in International Publication No. WO02006 / 011466, but is not limited to the production method.

  In the present invention, “free form” refers to a form in which Compound A does not form a salt.

  In the present invention, the “form I crystal” is a crystal form of a free compound A that does not contain a solvent or water in the crystal lattice, and 2θ (°) is approximately 12.5, 13.0, 14.9, 16.9, 18.0, 19.9. , 22.4, 22.7 and 23.1 show powder X-ray diffraction patterns including characteristic peaks. Further, when differential scanning calorimetry (hereinafter referred to as “DSC measurement”) is performed at a rate of temperature increase of 10 ° C./min, the peak temperature of the endothermic peak is shown between 137 ° C. and 147 ° C.

  In the present invention, the “form II crystal” is a DMSO hydrate of compound A containing dimethyl sulfoxide (hereinafter referred to as “DMSO”) in the crystal lattice, and 2θ (°) is approximately 5.4, 6.2, 11.1, 15.4. 1 shows powder X-ray diffraction patterns including characteristic peaks represented by 17.4 and 21.1.

  In the present invention, the “form III crystal” is a crystal form of the free compound A that does not contain a solvent or water in the crystal lattice, and 2θ (°) is approximately 5.8, 10.0, 10.9, 11.5, 11.9, and 20.5. The powder X-ray-diffraction pattern containing the characteristic peak represented by is shown.

  In the present invention, the “form IV crystal” is a crystal form of a free compound A that does not contain a solvent or water in the crystal lattice, and 2θ (°) is approximately 7.9, 11.7, 13.6, 17.9, 20.9, and 23.8. The powder X-ray-diffraction pattern containing the characteristic peak represented by is shown.

  In the present invention, the “sodium salt tetrahydrate crystal” is a crystal form of the sodium salt tetrahydrate of compound A, and 2θ (°) is represented by about 8.3, 10.7, 12.4, 14.2, 16.5 and 21.8. Shows a powder X-ray diffraction pattern including a characteristic peak.

  Each crystal is distinguished by thermal analysis such as powder X-ray diffraction measurement or differential scanning calorimetry (hereinafter referred to as “DSC”) measurement. The value of 2θ in powder X-ray diffraction measurements can vary in the range of plus or minus about 0.2 ° because instrumental differences and other factors can affect.

  Although the manufacturing method of each crystal form is described below, a manufacturing method is not limited to these, You may change the solvent to be used, temperature conditions, etc.

  Form I crystals can be obtained, for example, by dissolving compound A in acetonitrile or ethanol with heating and then cooling.

  Form II crystals can be obtained, for example, by dissolving Compound A in DMSO, lyophilizing the solution, and adding methylpentyl ketone and 1-chlorohexane to the resulting lyophilized product.

  Form III crystals can be obtained, for example, by dissolving compound A in methylpentyl ketone with heating and then adding diisopropyl ether.

  Form IV crystals can be obtained, for example, by dissolving compound A in ethyl acetate with heating and then adding diisopropyl ether.

  Sodium salt tetrahydrate crystals can be obtained, for example, by adding an aqueous sodium hydroxide solution containing compound A with sodium equivalent to compound A, water, and ethanol, dissolving them by heating, and then cooling.

  When the compound of the present invention is used as a MEK inhibitor, therapeutic agent or prophylactic agent for proliferative diseases, the compound of the present invention can be administered orally or parenterally. For oral dosage forms, tablets, capsules, granules, and powders are for oral dosage forms. For parenteral dosage forms, medicinal agents, eye drops, nasal drops, transdermal absorption agents, aerosols (“inhalants” And the like, and can be formulated using commonly used techniques.

  For example, oral preparations such as tablets, capsules, granules, powders are lactose, mannitol, starch, crystalline cellulose, light anhydrous silicic acid, calcium carbonate, calcium hydrogen phosphate and other excipients, stearic acid, magnesium stearate Lubricants such as talc, binders such as starch, hydroxypropylcellulose, hypromellose and polyvinylpyrrolidone, disintegrants such as carboxymethylcellulose, low-substituted hydroxypropylcellulose and calcium citrate, hypromellose, hydroxymethylcellulose, macrogol, silicone resin It can be adjusted by using a coating agent such as ethyl paraoxybenzoate, a stabilizer such as benzyl alcohol, a flavoring agent such as a sweetener, a sour agent, and a fragrance as necessary.

  Parenteral preparations such as injections and eye drops include isotonic agents such as sodium chloride, concentrated glycerin, propylene glycol, polyethylene glycol, potassium chloride, sorbitol, mannitol, sodium phosphate, sodium hydrogen phosphate, sodium acetate , Buffering agents such as citric acid, glacial acetic acid, trometamol, surfactants such as polyoxyethylene sorbitan monooleate, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil, stabilizers such as sodium citrate and sodium edetate , Benzalkonium chloride, paraben, benzotonium chloride, paraoxybenzoic acid ester, sodium benzoate, preservatives such as chlorobutanol, hydrochloric acid, citric acid, phosphoric acid, glacial acetic acid, sodium hydroxide, sodium carbonate, hydrogen carbonate sodium How pH adjusting agents, soothing agents such as benzyl alcohol, can be used depending on, for example, required a thickener such as hypromellose, is prepared.

  The dose of the compound of the present invention can be appropriately selected and used depending on symptoms, age, dosage form and the like. For example, the oral dosage is usually preferably 0.1 to 1000 mg, more preferably 1 to 30 mg per day. In the case of a parenteral preparation, it is preferably 0.1 to 1000 mg, more preferably 1 to 30 mg per day. This is preferably administered once or several times a day according to the symptoms.

  Preferred examples of the present invention will be described in more detail below, but the present invention is not limited to these examples.

[Production Example 1] 3,4-Difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1,2] oxazinane- Preparation of 2-ylmethyl) -benzamide (compound A)
Process 1
Synthesis of 2,3,4-trifluoro-5-iodo-benzoic acid

Acetic acid (1.2 L) and acetic anhydride (0.63 L) were added to a 5 L flask. While cooling with ice, sulfuric acid (1.6 L) was mixed in the same flask so as not to exceed 40 ° C. Further, at room temperature, I ₂ (70 g), activated MnO ₂ (70 g) and 2,3,4-trifluorobenzoic acid (120 g, 0.68 mol) were added as a solid with good stirring. While stirring the resulting black-violet suspension at 50 ° C., I ₂ (70 g) and MnO ₂ (70 g) were added after 2 to 3 hours, and I ₂ (33 g) and MnO ₂ ( An additional 37 g) was added. After 24 hours, the reaction mixture was added to ice (2 L).

  The purple organic layer obtained by extraction with methylene chloride three times (1.6 L + 1.0 L + 1.0 L) was extracted twice with 0.2N sodium thiosulfate aqueous solution (2.0 L + 1.0 L) for the purpose of removing excess iodine. ) When washed, the organic layer turned pale yellow. In order to remove acetic acid contained in the organic layer, it was washed 4 times with 1.0 L of water to obtain a transparent organic layer without turbidity. The organic layer was dried over anhydrous sodium sulfate (1.3 kg) and the solvent was distilled off under reduced pressure to give the target compound 2,3,4-trifluoro-5-iodo-benzoic acid (192 g, 94%) as a white powder. Obtained. If necessary, it can be washed with an EtOAc: Hexane = 1: 9 (v / v) solution.

¹ H-NMR (CDCl ₃ , 270 MHz) δ (PPM): 8.25 (1H, td, J = 6.6, 2.3 Hz)

Process 2
Synthesis of 2,3,4-trifluoro-5-vinylbenzoic acid

2,3,4-Trifluoro-5-iodo-benzoic acid (109 g, 0.36 mol), potassium vinyltrifluoroborate (68 g, 0.51 mol) obtained in Step 1, and Pd (dppf) Cl ₂ -CH ₂ Cl ₂ (15 g, 0.018 mol) was added as a solid to a 2 L flask, and then the vacuum-nitrogen purge was repeated twice for the purpose of bringing the entire experimental system to a nitrogen atmosphere. Methanol (0.80 L) and t-butylamine (95 mL, 0.91 mol) bubbled with nitrogen gas for 10 minutes for the purpose of deoxygenation were added. When heated to reflux with vigorous stirring under a nitrogen atmosphere, all of the compound dissolved and became a red solution. After 20 hours, the dark mixture with black precipitate was evaporated.

  Water (0.50 L), 1N aqueous sodium hydroxide solution (2.0 L) and methylene chloride (1.2 L) were added thereto and stirred to confirm that all of the compound was dissolved. The phosphorus ligand was removed to the methylene chloride layer. When washed twice with methylene chloride (1.0 L + 0.80 L), the dark color shifted to the organic layer and the aqueous layer containing the target compound became almost colorless. The aqueous layer whose pH was adjusted to about 1 by adding 12N hydrochloric acid (0.19 L) was extracted with ethyl acetate (1.2 L + 0.60 L + 0.80 L). The organic layer was washed with saturated brine (0.80 L), dried over anhydrous sodium sulfate (0.50 kg), and evaporated under reduced pressure to give the target compound 2,3,4-trifluoro-5-vinylbenzoate as a pale yellow solid. The acid (72 g, 98%) was obtained.

¹ H-NMR (CDCl ₃ , 270 MHz) δ (PPM): 5.54 (1H, d, J = 11.2 Hz), 5.92 (1H, d, J = 17.8 Hz), 6.78 (1H, dd, J = 17.8, 11.2 Hz), 7.95 (1H, td, J = 7.6, 2.6 Hz)
EIMS m / z 202 (M + ・)

Process 3
Synthesis of 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -5-vinyl-benzoic acid

  2-Fluoro-4-iodo-phenylamine (108 g, 0.46 mol) was added to a dried glass 3 L flask as a solid, and 2,3,4-trifluoro-5-vinylbenzoic acid obtained in Step 2 ( 77 g, 0.38 mol) was added to the dropping funnel as a solid. In order to make it nitrogen atmosphere (dehydration, deoxygenation), vacuum-nitrogen purge was repeated twice. Dehydrated / deoxygenated THF was added to a 3 L flask, 0.65 L, and 0.30 L was added to a dropping funnel to dissolve the two kinds of raw materials. A 1M THF solution of LiHMDS (1.1 L, 1.1 mol) was added to another dropping funnel and dropped into a 3 L flask cooled to −78 ° C. over 45 minutes, resulting in a pale yellow suspension. Subsequently, when 2,3,4-trifluoro-5-vinylbenzoic acid THF solution in the dropping funnel was dropped over 10 minutes, the color turned orange. When the reaction temperature was gradually raised and the temperature reached 0 ° C. 3 hours after the completion of the dropwise addition, a dark colored solution was formed. After further stirring for 2 hours at room temperature, the reaction was stopped by adding 0.50 L of water while cooling with ice, and the color turned orange.

  Furthermore, 1.2 L of 2N hydrochloric acid was added after 5 minutes. After obtaining the THF layer by separation, the aqueous layer was further extracted with ethyl acetate (2 × 0.80 L). The red organic layers were combined, washed with saturated brine (0.80 L), dried over anhydrous sodium sulfate (0.50 kg), and evaporated to give 178 g of an orange solid. The solid was washed with methylene chloride (total 0.60 L) to give the target compound 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -5-vinyl-benzoic acid (150 g) as a white solid. , 0.36 mol, 94%).

¹ H-NMR (CD ₃ OD, 270 MHz) δ (PPM): 5.41 (1H, d, J = 11.5 Hz), 5.86 (1H, d, J = 17.2 Hz), 6.76 (1H, td, J = 8.6, 5.6 Hz), 6.79 (1H, dd, J = 17.2, 11.5 Hz), 7.41 (1H, m), 7.48 (1H, dd, J = 10.6, 2.0 Hz), 8.05 (1H, dd, J = 7.9, 2.0 Hz)
ESI (LC / MS positive mode) m / z 420 (M + H)

Process 4
Synthesis of 2- (2-hydroxy-ethoxy) -isoindole-1,3-dione

  N-hydroxyphthalimide (500 g, 3.07 mol) was added to a three-necked flask (5 L) equipped with a reflux apparatus under a nitrogen atmosphere, and anhydrous acetonitrile (1.0 L) was added and stirred well to obtain a suspension. The reaction vessel was cooled in an ice bath, and when the internal temperature reached 15 ° C, triethylamine (514 mL, 3.68 mol) was added dropwise over about 10 minutes. The suspension became a dark red solution. Subsequently, 2-bromoethanol (260 mL, 3.678 mol) was added dropwise, the ice bath was removed, and the temperature was returned to room temperature. The reaction vessel was heated in an oil bath at an internal temperature of 80 ° C. for 8 hours. The insoluble matter precipitated after returning to room temperature was filtered off with a glass filter, washed with about 1.0 L of ethyl acetate, and the filtrate and washings were combined and concentrated under reduced pressure.

  Water (1.5 L) was added to the reddish brown oily residue and extracted with ethyl acetate (3 × 2.0 L). Each extracted organic layer was washed with saturated brine (3 × 1 L) and dried over anhydrous magnesium sulfate (1.5 kg). The solvent was distilled off under reduced pressure to obtain an off-white solid (599.5 g, crude yield 94%). EtOAc-hexane (1.2 L) was added to this and triturated well. The white solid obtained by filtration was washed with EtOAc-hexane (0.5 L) and hexane (1.0 L) in this order, and dried under reduced pressure. The target compound 2- (2-hydroxy-ethoxy) -isoindole-1,3-dione (520 g, 82%) was obtained as a white solid.

¹ H-NMR (CDCl ₃ , 270MHz) δ (PPM): 3.46 (1H, t, J = 7.1 Hz), 3.81 (2H, m), 4.32 (2H, t, J = 4.5 Hz), 7.79 (2H, m), 7.87 (2H, m)
LC-MS: m / z = 208 (M + H)

Process 5
Synthesis of 2-aminooxy-ethanol

  To a 5 L flask was added 2- (2-hydroxy-ethoxy) -isoindole-1,3-dione (555 g, 2.679 mol) obtained in Step 4, and dissolved in methylene chloride (2.0 L) with stirring. While the reaction vessel was cooled in an ice bath, methyl hydrazine (142 mL, 2.68 mol) was added dropwise so that the internal temperature did not exceed 20 ° C. As the reaction progressed, a white insoluble material appeared. After completion of the dropwise addition, the mixture was stirred in an ice bath for about 15 minutes. The precipitated insoluble matter was filtered off with a glass filter, and the white insoluble matter on the filter was washed with dichloromethane (2 x 400 mL). Combined and evaporated under reduced pressure. The yellow oily residue (230 g) was distilled under reduced pressure (51 to 54 ° C./2 mmHg) to obtain the target compound 2-aminooxy-ethanol (162 g, 73%) as an unemployed transparent liquid.

¹ H-NMR (DMSO-D ₆ , 270 MHz) δ (PPM): 3.51 (4H, m), 4.53 (1H, t, J = 5.0 Hz), 5.93 (2H, br. S)
GC-MS: m / z = 77 (M +)

Step 6
Synthesis of 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5-vinyl-benzamide

3,4-Difluoro-2- (2-fluoro-4-iodo-phenylamino) -5-vinyl-benzoic acid (180 g, 0.429 mmol) and 3-hydroxy-4-oxo-3 obtained in step 3 , 4-Dihydro-1,2,3-benzotriazine (77.0 g, 0.472 mmol) dissolved in N, N-dimethylformamide (2 L), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride Salt (87.2 g, 0.455 mol) was added, and the mixture was stirred at room temperature for 2 hours. After confirming the formation of active ester and disappearance of raw materials by LC-MS, the reaction system was cooled to 0 ° C.
2-Aminooxy-ethanol (39.7 g, 0.515 mol) obtained in step 5 was added and stirred at 0 ° C. for 2 hours. LC-MS confirmed the formation of the condensate and the disappearance of the active ester.

  To the reaction solution were added 0.3N hydrochloric acid (2 L) and saturated brine (500 ml), and the resulting mixture was extracted with ethyl acetate (2 × 2 L). Combine the organic layers, wash with 0.3N hydrochloric acid (2L), saturated aqueous sodium bicarbonate (2 x 2L), saturated brine (2L), dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain the target compound as a white solid. 3,4-Difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5-vinyl-benzamide (179 g, 87%) was obtained.

¹ H-NMR (DMSO-D ₆ ) δ (PPM): 3.57 (2H, m), 3.85 (2H, t, J = 4.6 Hz), 4.71 (1H, br.s), 5.49 (1H, d, J = 11.2 Hz), 5.97 (1H, d, J = 17.6 Hz), 6.71 (1H, td, J = 8.8, 4.4 Hz), 6.78 (1H, dd, J = 17.6, 11.2 Hz), 7.36 (1H, d , J = 8.3 Hz), 7.57 (1H, dd, J = 11.0, 1.7 Hz), 7.64 (1H, d, J = 6.8 Hz), 8.75 (1H, br.s), 11.89 (1H, br.s) .
ESI (LC / MS positive mode) m / z 479 (M + H)

Step 7
Synthesis of 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -5-formyl-N- (2-hydroxy-ethoxy) -benzamide

3,4-Difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5-vinyl-benzamide (179 g, 0.374 mol) obtained in Step 6 Dissolved in a mixed solvent of THF (950 ml) and water (950 ml), cooled to 0 ° C., and further cooled with N, N-dimethylacetamide (13.9 mL, 0.149 mol) and microencapsulated osmium oxide (Wako Pure Chemical Industries, 4.75 g, 1.87 mmol, 0.005 eq.) Was added, and the mixture was stirred at 0 ° C. for 30 minutes, and then NaIO ₄ (320 g, 1.49 mol) was added in three portions at 5 to 10 minute intervals. The mixture was stirred at 0 ° C. overnight (14 hours). The reaction solution was filtered using Nutsche, and insoluble matters on the filter paper were washed with THF (1 L).

  The filtrate and washings were combined and diluted with ethyl acetate (2 L), washed with 0.2 M aqueous sodium thiosulfate (2 L) and 1.0 N hydrochloric acid (30 ml), and the aqueous phase was separated. Further, the organic layer was washed with 0.2 M aqueous sodium thiosulfate solution (2 L) and saturated brine (500 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product (158 g). Recrystallization from ethyl acetate and yellow powder as the target compound 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -5-formyl-N- (2-hydroxy-ethoxy) -benzamide ( 101 g, 56%).

¹ H-NMR (DMSO-D ₆ , 270 MHz) δ (PPM): 3.59 (2H, t, J = 4.6 Hz), 3.85 (2H, m), 4.73 (1H, br.s), 6.99 (1H, td , J = 8.8, 3.4 Hz), 7.48 (1H, d, J = 8.8 Hz), 7.66 (1H, dd, J = 10.5, 1.7 Hz), 7.86 (1H, d, J = 6.8 Hz), 9.59 (1H , br.s), 10.02 (1H, s), 12.10 (1H, br.s).
ESI (LC / MS positive mode) m / z 481 (M + H)

Process 8
Synthesis of 4- (1,3-dioxo-1,3-dihydro-isoindol-2-yloxy) -butyric acid ethyl ester

  4-Bromo-n-butyric acid ethyl ester (120 g, 615 mmol) was dissolved in dimethylformamide (900 ml), and N-hydroxyphthalimide (120 g, 738 mmol) and N, N-diisopropylethylamine (214 mL, 1.23 mol) was added, and the mixture was stirred at 80 ° C. overnight. The reaction solution was poured into a saturated aqueous ammonium chloride solution, and the resulting mixture was extracted with ethyl acetate (2 L + 1 L). The organic layers were combined, washed with saturated brine (3 x 500 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 4- (1,3-dioxo-1,3-dihydro-isoindole-2 as a crude compound. -Iyloxy) -butyric acid ethyl ester (149 g) was obtained.

  If necessary, this crude compound can be purified by silica gel column chromatography (n-hexane / ethyl acetate = 2: 1).

¹ H-NMR (CDCl ₃ , 270 MHz) δ (PPM): 1.28 (3H, t, J = 6.9 Hz), 2.11 (2H, q, J = 6.9 Hz), 2.64 (2H, t, J = 7.3 Hz) , 4.17 (2H, q, J = 7.3 Hz), 4.27 (2H, t, J = 6.3 Hz), 7.72-7.79 (2H, m), 7.81-7.89 (2H, m).

Step 9
Synthesis of 4-aminooxy-N-methyl-butyramide

The original compound of 4- (1,3-dioxo-1,3-dihydro-isoindol-2-yloxy) -butyric acid ethyl ester obtained in step 8 was dissolved in methanol (500 ml) and methylamine was obtained at room temperature. (40% methanol solution, 1000 ml) was added and stirred overnight at 60 degrees. The reaction solution was concentrated under reduced pressure, methylene chloride was added to the resulting residue, and the precipitated solid was filtered and washed with methylene chloride. The filtrate and washings were combined and concentrated under reduced pressure. When this operation was repeated three more times, no solid precipitated even when methylene chloride was added to the residue. The oily residue (approx. 150 g) was purified by silica gel column chromatography (silica gel 1.5 kg, CH ₂ Cl ₂ / MeOH = 9: 1 ~ 7: 1) and 4-aminooxy-N-methyl as a pale yellow syrup -Butyramide (57.2 g, 70% yield over 2 steps) was obtained.

¹ H-NMR (CDCl ₃ , 270 MHz) δ (PPM): 1.94 (2H, q, J = 6.3 Hz), 2.25 (2H, t, J = 6.9 Hz), 2.81 (3H, d, J = 4.6 Hz) , 3.70 (2H, t, J = 5.9 Hz).

Step 10
(E) -3,4-Difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5-{[(3-methylcarbamoyl-propoxyimino) -methyl Synthesis of ] -benzamide

  34.7 g (0.263 mol) of 4-aminooxy-N-methyl-butyramide obtained in Step 8 was dissolved in a mixed solvent (1.8 L) of tetrahydrofuran / methanol (3: 1) to obtain 3,3 4-Difluoro-2- (2-fluoro-4-iodo-phenylamino) -5-formyl-N- (2-hydroxy-ethoxy) -benzamide (90.0 g, 0.187 mol) was added and stirred for 2 days. The reaction solution was concentrated under reduced pressure until a slight amount of solvent remained, and then acetonitrile (1.1 L) was added to form a precipitate. The precipitate is filtered, washed with acetonitrile, and then dried under vacuum to obtain the target compound (E) -3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy ) -5-{[(3-Methylcarbamoyl-propoxyimino) -methyl] -benzamide (111 g, 80%) was obtained.

¹ H-NMR (DMSO-D ₆ , 270 MHz) δ (PPM): 1.88 (2H, qui, J = 7.6 Hz), 2.17 (2H, t, J = 6.9 Hz), 2.56 (3H, d, J = 4.6 Hz), 3.57 (2H, br.q, J = 4.6 Hz), 3.83 (2H, t, J = 4.6 Hz), 4.14 (2H, t, J = 6.3 Hz), 4.73 (1H, t, J = 5.6 Hz), 6.80 (1H, td, J = 8.9, 4.0 Hz), 7.40 (1H, br.d, J = 8.6 Hz), 7.61 (1H, dd, J = 10.9, 2.0 Hz), 7.68 (1H, br d, J = 5.6 Hz), 7.77 (1H, br q, J = 4.6 Hz), 8.26 (1H, s), 8.87 (1H, br.s), 11.99 (1H, br.s).
ESI (LC / MS positive mode) m / z 595 (M + H)

Step 11
3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1,2] oxazinan-2-ylmethyl)- Synthesis of benzamide (compound A)

  (E) -3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5-{[(3-methylcarbamoyl) obtained in step 10. -Propoxyimino) -methyl] -benzamide (20.0 g, 33.7 mmol) suspended in methylene chloride (600 ml) and borane-pyridine complex (13.6 ml, 0.135 mol) and dichloroacetic acid (16.6 ml, 0.202) at room temperature mol) was added. After stirring at room temperature for 14 hours, methylene chloride was removed under reduced pressure using a rotary evaporator. The residue was diluted with 1,2-dichloroethane (600 ml), the mixture was stirred at 60 ° C. for 8 hours, and then the reaction solution was filtered. The residue obtained by concentrating the filtrate was diluted with ethyl acetate (2 L) and washed with water (1 L), saturated aqueous sodium hydrogen carbonate (1 L), and saturated brine (1 L) in this order. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure.

The residue was roughly purified by silica gel column chromatography (silica gel 700 g, CH ₂ Cl ₂ / MeOH = 20: 1), and 3,4-difluoro-2- (2-fluoro-4-iodo as a crude compound obtained -Phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1,2] oxazinan-2-ylmethyl) -benzamide was recrystallized from methylene chloride / pentane (2: 3) It was. Combine the primary crystal (13.7 g) and the secondary crystal (2.8 g), and further to silica gel column chromatography (silica gel 640 g, AcOEt / MeOH (30: 1) → CH ₂ Cl ₂ / MeOH (4: 1)) The desired compound 3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1, 2] Oxazinan-2-ylmethyl) -benzamide (Compound A) (15.8 g, 83%) was obtained.

¹ H-NMR (CD ₃ OD, 400 MHz) δ (PPM): 2.10 (2H, qui, J = 6.8 Hz), 2.53 (2H, t, J = 6.8 Hz), 3.72 (2H, t, J = 4.4 Hz ), 3.92 (2H, t, J = 4.4Hz), 4.04 (2H, t, J = 6.8Hz), 4.86 (2H, s), 6.61 (1H, ddd, J = 8.8, 8.8, 3.9 Hz), 7.33 -7.36 (1H, m), 7.41 (1H, dd, J = 7.0, 1.7 Hz), 7.45 (1H, dd, J = 10.3, 2.0 Hz).
ESI (LC / MS positive mode) m / z 566 (M + H)

Powder X-ray diffraction measurement
The measurement conditions of powder X-ray diffraction measurement using Cu Kα (1.54060 に) as the X-ray source are shown below.
Measurement conditions for free form I, II, III, and IV crystals Measurement device: X'Pert-Pro MPD (manufactured by PANalytical)
Counter cathode: Cu
Tube voltage: 40 kV
Tube current: 40 mA
Scanning range: 3 to 35 °
Sampling width: 0.025 °

Measurement conditions of sodium salt tetrahydrate crystal <br/> Measuring device: D8 Discover with GADDS CS diffractometer (Bruker AXS)
Counter cathode: Cu
Tube voltage: 45kV
Tube current: 40 mA
Scanning range: 5-25 °
Sampling width: 0.02 °

DSC measurement
The measurement conditions for DSC measurement are shown below.
Measuring device: DSC6200R (manufactured by SII NanoTechnology)
Temperature increase rate: 10 ° C / min Atmosphere: Under nitrogen stream Sample container: Simple sealed sample container made of aluminum

[Example 1] Preparation of free form Form I of Compound A Compound A powder (3.2 g) was dissolved in acetonitrile (10.7 mL) at 120 ° C, cooled to room temperature, and stirred for 45 minutes. The precipitated crystals were filtered and dried to obtain a free form I crystal of Compound A as a white crystalline solid (2.8 g). The powder X-ray diffraction pattern of the crystals is shown in FIG. Table 1 shows characteristic diffraction angles of peaks.

A DSC chart is shown in FIG.
Endothermic peak: around 143 ° C.

[Example 2] Preparation of free form II crystal of Compound A Compound A powder (970 mg) was dissolved in DMSO (5 mL), and this solution was freeze-dried at -20 ° C for 3 days. Methyl pentyl ketone (1.6 mL) and 1-chlorohexane (3.2 mL) were added to the resulting lyophilized product, and crystals were obtained by shaking and stirring at room temperature for 2 days. The precipitated crystals were filtered and dried to obtain free form II crystals as a white crystalline solid (760 mg). The powder X-ray diffraction pattern of the crystals is shown in FIG. Table 2 shows characteristic peak diffraction angles.

A DSC chart is shown in FIG.
Endothermic peak: around 81 ° C.

[Example 3] Preparation of free form III crystal of compound A Compound A powder (980 mg) was heated and dissolved in methylpentyl ketone (28 mL), diisopropyl ether (137 mL) was added, and the mixture was allowed to stand at 5 ° C for 3 days. To obtain crystals. The precipitated crystals were filtered and dried to obtain a white crystalline solid (810 mg) which was a free form III crystal. The powder X-ray diffraction pattern of the crystals is shown in FIG. Table 3 shows characteristic peak diffraction angles.

A DSC chart is shown in FIG.
Endothermic peak: around 124 ° C.

[Example 4] Preparation of free form IV crystal of compound A Compound A powder (980 mg) was dissolved in ethyl acetate (39 mL) with heating, diisopropyl ether (98 mL) was added, and the mixture was allowed to stand at 5 ° C for 3 days. As a result, crystals were obtained. The precipitated crystals were filtered and dried to obtain a white crystalline solid (830 mg) which was a free form IV crystal. The powder X-ray diffraction pattern of the crystals is shown in FIG. Table 4 shows characteristic diffraction angles of peaks.

A DSC chart is shown in FIG.
Endothermic peak: around 143 ° C

[Example 5] Preparation of sodium salt tetrahydrate crystals of compound A To a powder of compound A (500 mg), 2N aqueous sodium hydroxide solution (0.44 mL), water (10 mL) and ethanol (2 mL) were added and heated. After dissolving, crystals were obtained by cooling to room temperature. The precipitated crystals were filtered and dried under air to obtain a white crystalline solid (425 mg) as sodium salt tetrahydrate crystals. The powder X-ray diffraction pattern of the crystals is shown in FIG. Table 5 shows characteristic diffraction angles of peaks.

A DSC chart is shown in FIG.
Endothermic peak: Peak derived from dehydration at 90 ° C and 100 ° C, Peak derived from melting of crystal at around 226 ° C

[Test Example 1] Thermodynamic stability test between polymorphs of the free form A thermodynamic stability test was conducted using the free form I crystal, free form III crystal, and free form IV crystal of the present invention. . Each crystal prepared by the method of Example 1 and Examples 3 and 4 was used as a sample. Free form I (2 mg) and free form III (2 mg) were taken in a vial and 20 μL of solvent was added. Water and 50% ethanol were used as the solvent. Similarly, free form I and free form IV were also taken in vials and the same procedure was performed. After each vial was stirred for 1 week at room temperature, powder X-ray diffraction measurement was performed on the crystals in the vial to confirm the crystal form. The results are shown in Table 6. Under any condition, the crystals obtained were free form I crystals. From this, it was shown that the free form I is thermodynamically more stable than the free form III and free form IV.

[Test Example 2] Storage stability test A storage stability test was conducted using the free form I crystal and sodium salt tetrahydrate crystal of the present invention. Each crystal prepared by the method of Example 1 and Example 5 was used as a sample. Each sample was stored in a desiccator containing silica gel (drying conditions) and a desiccator containing saturated sodium chloride aqueous solution (humidification conditions: relative humidity of about 75%), and each desiccator was left in a high-temperature bath at 80 ° C. Two weeks later, powder X-warp diffraction measurement of each sample was performed to confirm the crystal form. The results are shown in Table 7. Sodium salt tetrahydrate was dehydrated under dry conditions and deliquescent under humidified conditions to become amorphous. On the other hand, the free form I crystals were stable with no transition under dry or humid conditions.

[Test Example 3] Light Stability Test A storage stability test was performed using the free form I crystal and sodium salt tetrahydrate crystal of the present invention. Each crystal prepared by the method of Example 1 and Example 5 was used as a sample. Each sample was placed in a transparent glass container and allowed to stand in a light stability tester (LABONIC LA110, ETAC). A D65 lamp (FL20S / D-EDL-D65, Toshiba) was used as a light source, and the light was irradiated with a total illumination of 864 × 10 ³ lux · hr and a total near-ultraviolet irradiation energy of 233 W · h / m ² . Moreover, the sample weighed similarly as a sample before light irradiation was light-shielded and preserve | saved. Each sample before light irradiation and after light irradiation was quantified by HPLC, and the residual ratio by storage was determined.

  Each sample was quantified by the following procedure. A sample (about 1 mg) was weighed and dissolved in ethanol (2.5 mL) to obtain a sample solution. 2 μL of each sample solution was measured under the following HPLC conditions.

HPLC condition equipment: Waters HPLC (2790 separation module)
Column: CAPCELL PAK C18 UG120, 3mmφ × 50mm (Shiseido)
Column temperature: 30 ° C
Mobile phase A: water containing 0.1% trifluoroacetic acid Mobile phase B: acetonitrile containing 0.1% trifluoroacetic acid Flow rate: 0.5 mL / mL
Mobile phase condition: 38% B
Sample cooler temperature: 5 ° C
Detection wavelength: 230 nm
Measurement range: 17 minutes after sample solution injection

  From the peak area derived from the compound having a retention time of about 4.9 minutes, the residual rate after light irradiation of each sample was calculated according to the following formula.

  The results are shown in Table 8. The decomposition of sodium salt tetrahydrate crystal proceeded by light irradiation, and the residual rate decreased to 82.9%, but the free form I crystal was stable with a residual rate of 99.4%.

  From the results of Test Example 3, it is suggested that among the crystalline polymorphs of the free form, Form I is more stable than Forms III and IV. Furthermore, the results of Test Examples 4 and 5 suggest that the free form I crystal is more stable than the sodium salt tetrahydrate crystal. In addition, since the free form II contains DMSO in the crystal lattice, it is not useful as a pharmaceutical due to toxicity derived from DMSO. From these viewpoints, the free form I crystal of the present invention can stably provide a pharmaceutical product without stabilization by special pharmaceutical treatment, special packaging, special storage conditions, or the like.

  The present invention is a crystal of compound A that is an excellent MEK inhibitor and is useful as a pharmaceutical having excellent storage stability that does not need to be stabilized by special pharmaceutical treatment, special packaging, special storage conditions, or the like. And crystal polymorphs thereof.

Claims (11)

  3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1,2] oxazinan-2-ylmethyl)- Benzamide crystals.   3,4-difluoro-2- (2-fluoro-4-iodo-phenylamino) -N- (2-hydroxy-ethoxy) -5- (3-oxo- [1,2] oxazinan-2-ylmethyl)- 2. The crystal according to claim 1, wherein the benzamide is in a free form.   3. The crystal according to claim 2, comprising peaks having diffraction angles represented by 2θ of 14.9, 16.9, and 23.1 ° (± 0.2 °) in a powder X-ray diffraction pattern.   4. The crystal according to claim 3, wherein the powder X-ray diffraction pattern further includes peaks having diffraction angles represented by 2θ of 12.5, 18.0 and 22.7 ° (± 0.2 °).   5. The crystal according to claim 4, further comprising peaks having diffraction angles represented by 2θ of 13.0, 19.9, and 22.4 ° (± 0.2 °) in the powder X-ray diffraction pattern.   6. The crystal according to claim 5, wherein the peak temperature of the endothermic peak in DSC measurement (temperature increase rate: 10 ° C./min) is from 137 ° C. to 147 ° C.   2. The crystal according to claim 1, comprising peaks having diffraction angles represented by 2θ of 5.4, 6.2, 11.1, 15.4, 17.4 and 21.1 ° (± 0.2 °) in a powder X-ray diffraction pattern.   2. The crystal according to claim 1, comprising peaks having diffraction angles represented by 2θ of 5.8, 10.0, 10.9, 11.5, 11.9 and 20.5 ° (± 0.2 °) in a powder X-ray diffraction pattern.   2. The crystal according to claim 1, comprising a peak having diffraction angles represented by 2θ of 7.9, 11.7, 13.6, 17.9, 20.9, and 23.8 ° (± 0.2 °) in a powder X-ray diffraction pattern.   2. The crystal according to claim 1, comprising peaks having diffraction angles represented by 2θ of 8.3, 10.7, 12.4, 14.2, 16.5, and 21.8 ° (± 0.2 °) in a powder X-ray diffraction pattern.   The pharmaceutical composition containing the crystal | crystallization in any one of Claims 1-11.

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