SPECIFICATION
Crystals of Phenylalanine Derivatives and Production Methods Thereof
Technical Field of the Invention
The present invention relates to crystals of phenylalanine derivatives that have the specific structural formula find production methods thereof. Particularly, it relates to a-typo, retype, E-typo, n-type, and fHypo crystals thereof.
Background of the Invention
A compound of the following formula (I) (lutntliiaflw roiorrod to as a compound (0) or pharmaceutically acceptable suits thnreof aro the compounds which have an a 4 integrin inhibiting activity and are useful as agents for treating inflammatory bowel diseases and the 11 lie. Though they can be produced in accordance with the description of Patent literature 1, there is no disclosure on "crystals" of the compound (I) or phnrmaeoutleally acceptable salts thereof in the publication.
Generally, when preserving drug substances or processing or preserving
I preparations, the amorphous or noncrystalline solid drug substances are unstable in the environmental conditions such as temperature, humidity, air, and the like. Therefore, it can be problematic in developing highly-pure pharmaceutical compositions. Further, since the amorphous or noncrystalline solid drug substances have dogradability by moisture absorption, solvents that
i can be used in processing preparations are limited to anhydrides and, therefore, it can cause the increase in preparation costs. Mesldes, the drug substances have to be those that can be produced on the industrial scale.

[Patent Literature l] WO02/16329
Disclosure of Iho Invention
Tbo object of the present invention is to provide crv/italM ol' the compound (l) that are excellent in preservation stability or moisture resistance.
Tiie further object of the present invention in to provide crystals of the compound (I) that can be produced on the industrial scale.
The inventors have thoroughly studied the above problems to solve them and researched crystal forma of the compound (I) or phannacoutieally acceptable salts thereof. Then, they have surprisingly fovind that tint compound (I) itself that does not form the salts is excellent in stability or crystallization, and they have also found five novel crystal forms to solve the above problems from several crystal forms of the compound (I). The present invention has boon completed based on these findings.
Namely, the present invention is mentioned as follows.
(1) A crystal of the compound of the formula (I).

(2) The crystal according to above (l), which is an rrtype, ?'-typo, E "type, r•type, or 6 -type.

(3) A method for producing the a -type crystal which comprises Iho steps of dissolving the compound of the formula (I) in n good solventCs) containing at least one land selected from the group consisting of ncotonitrile, dichloromethane, tetrahydrofuran, acetone, dimethylsulfoxido and chloroform, or in n mixed solvent of acctonitrile-water, or in a mixed solvent of acetonilrile-dimethylformamide; and cooling down the mixture to 0 to 3()°C to crystallize.
(4) A method for producing the a-type cryMtr.il which coinpi'iHOH the stops of dissolving tho compound of the formula (I) in a good NOIVOIII.(N) and then adding a poor solvont(s) thereto to crystallize, wherein the comhination of the good solvent(H) and the poor solvent(s) is either one el' dlmolhylMulfoxido-toluone, dhnotliylfnniuimide-diothylether, dliiinlhylform amide-toluene, chloroform-nth nnol, chloroform-toluene, chloroform-diothylether, dichloromothnne-diothylethor, totnihydrofurmrwater, tetrahydrofurnircyclohexane, acetone-water, aeotottiti'ilo-wator or dimothylformamide-acetonitrile.
(5) A method for producing the a-type crystal which comprises the stops of suspending the compound of the formula (I) in aool.ouitrilo, a mixed solvent of acetonitrile-water or a mixed solvent of acetonilrilo-dimethylformamide; and stirring the mixture at 0 to 40°C to crystallize.
(6) A method for producing the T"type crystal which comprises the steps of dissolving the compound of the formula (I) in dhmdhyl form amide and then cooling down the mixture to 0 to 30°C to crystallize.
(7) A method for producing the T'-type crystal which comprises the steps of dissolving the compound of the formula (I) in a good sol von 1(H) and then adding a poor solvent(s) thereto to crystallize, wherein the comhinntion of the good solventCs) and the poor solvent(s) is dimethylforinamlde-wator.

(8) A method for producing the z -type crystal which comprises tho .stop.s of dissolving the compound of tho formula (l) in a good solvents) and than adding a poor solveut(s) thereto to crystallize, wherein tho cornhinatinn of tho good solvent(s) and the poor solvont(s) is either one of dlchmrnniothnne-ethnnol or dimethylsulfoxide-diethylether.
(9) A method for producing the r\ -type crystal which comprises (lie steps of: dissolving the compound of the formula (I) or a hydrochloride of the compound of the formula (1) in an alcohol solution having I to (I carhon atoms containing hydrogen chloride! and then neutralizing the mixture with a hase(s) to crystallize.
(10) A method for producing the 0-type crystal which comprises the steps of: suspending the compound of the formula (I) in a mixed solvent of dimethylformamide-acetonitrile or in a mixed solvent of aeetonitrilo-water; and stirring the mixture at 40°C or higher to crystallize.
Brief Description of the Drawings
Fig. I shows the diagram of the powder X-ray diffraction pattern on the novel a -type crystal of the present invention (Tim horizontal axis indicates the diffraction angle 2 61° ]; and the vertical axis indicates the strength I.CPS].)
Fig. 2 shows the diagram of the powder X-ray diffraction pattern on tho novel T "type crystal of the present invention (The horizontal aids indicates the diffraction angle 2 6 [" ]', and the vertical axis indicates the strength |(Tf\S|.)
Fig. I) shows the diagram of the powder X-ray diffraction pattern on the novel E -type crystal of the present invention (The horizon till axis indicates the diffraction angle 2d[° ]', and the vertical axis indicates tho strength |CI\S|.)
Pig. 4 shows the diagram of the powder X-ray diffraction pattern on the novel 77 -type crystal of the present invention (The horizontal axis indicates the

diffraction angle 2 6 [° ]; and the vertical axis indicates the strength [(IPS].)
Fig. 5 .shows the diagram of the powder X-ray diffraction pattern on the novel 8 -type crystal of the present invention (The horizontal axis indicates the diffraction angle 2 81° ]; and the vertical axis indicates 1,1 to strength [OPS].)
Fig. 6 shows the diagram of the powder X-ray diffraction pattern on the publicly-known compound (a hydrochloride of the compound of the formula (I)) indicated in Comparative Example 1 (The horizontal axis indicates the diffraction angle 2 8 [° ]; and the vertical axis indicates the strength ICPSI.)
Fig. 7 shows the infrared, absorption spectrum of the (V -type crystal.
Fig. 8 shows the DSC patterns of the w typo, y type, t: type and f/type crystals,
Fig, 9 shows the temperature changes of the powder X ray diffraction patterns of the a -type, y -type, e -type and ?/ type crystals.
Fig. 10 shows the T-G curves (T: temperature |"C|, Q: the relative value of Gibbs free energy) of the a -type, f-type, e -type, 1\ type, and 8 -typo crystals.
Fig. U shows the vapor adsorption isotherms of the Of-typo, f-typo and £ -type crystals (The horizontal axis indicates the relative humidify |%]/]0(); and the vertical axis indicates the water adsorption amount [%].)
Fig. 12 shows the vapor adsorption isotherm of the ?/ -type crystal (The horizontal axis indicates the relative humidity |%|/1()(); and the vertical axis indicates the water adsorption amount [%].)
Best Mode for Carrying out the Invention
The definitions or examples in the present specification are indicated as follows.
"Tim compound of the formula (I)" is described in Example l!.!(i of Patent Literature I.

"Crystal" mean a solid substance thai. gives n characteristic (l.i:fl'raction diagram in the ])owder X-ray analysis and usually menus crystals or crystalline solid.1) h'urther, the mixtures of crystals and amorphous are acceptable. In such a case, the crystals may be contained in the miliHlnnllal ratio.
The (V type crystal" means the crystal wltereln the ponies exist at the diffraction angles (2 0) of G.2, 1.0.2, 10.7, 10.8, MO, I'M, 10.0, 10.2 and 21.7 in the powder X-ray diffraction pattern. Particularly, it moans the crystal wherein the peaks exist at the diffraction angles (2 0) of 10.7, 10.8, 14.0, 1(1.2 and 21.7.
"The y -type crystal" means the crystal wherein the peaks exist at the diffraction angles (20) of 7.2, 8.1, 10.3, 10.9, 14.5, 15.1, 10.4, 17.3, 18.3, 19.4, and 23.3 in the powder X-ray diffraction pattern.
"The E -type crystal" means the crystal wherein the peaks exist at the diffraction angles (2 0) of 5.4, 6.9, 8.3, 10.8, 11.1, 12.8, 10.1, 17.7, 21.G 23.4, 24,5, and 25.1 in the powder X-ray diffraction pattern.
"The n -type crystal" moans the crystal wherein the peaks exist at the diffraction angles (2 0) of 9.7, 12.2, 12.8, 14.9, 15.0, 10.9, 18.5 and 20.4 in the powder X-ray diffraction pattern.
"The 0 -type crystal" means the crystal wherein the peaks exist at the diffraction angles (2 0) of 5.7, 10.3, 11.5, 13.1), 10.5, 18.5, 20.0 and 21.0 in the powder X-ray diffraction pattern. Particularly, it moans the crystal wherein the peaks exist at.the diffraction angles (2 6) of U .5, I III), I«.5, 20.0 and 21.0.
In (he production methods of the present invention, in addition to using amorphous or noncrystalline solids as the compound (I), the raw material, the amorphous or noncrystalline solids may be used to once proparo crystals, and then other crystals may be produced using the obtained crystals in accordance with the other production methods of the present invention.
The used crystallization processes include, for example, the crystallization by

cooling, the crystallization with a poor solvont(s), the crystallization by suspension, the crystallization by neutralization, and tin) crystal I iza lion by concentration. Any process can be conducted if the subject compound is dissolved or suspended to a crystallizing solvent(s) to crystallize. In the crystallization, a seed crystal(s) is preferably added to the mixture. Meanwhile, the crystallization by cooling and the crystallization with a poor solvont(s) may be combined together.
Tin* crystallizing solvents are those that are um.in.llv known as usable crystallizing solvents. They may be one kind or a mixed solvent(s) of several crystal I izi ng solvents.
As for the mixed solvents of the several crystallizing solvents, it is possible to use the mixed solvent(s) wherein the solvent thai dissolves the subject compound well (a good solvent) and the solvent that is soluble in the good solvent but hardly dissolves the subject compound (a poor solvent) are mixed in suitable quantities, it is also possible to use several solvents as the good solvent mid those as the poor solvent. In that case, it is preferable that they equally intermingle with each other.
The solvents that dissolve the subject compound (the compound of the formula CO) well can be used as "the good solvents." They include acotonitrile, tetrahydrofuran, acetone, chloroform, dichloromothnno, dlmothylformamide, fbrmamide, and dimethylsulfoxide.
The solvents that are soluble in the above good solvents but hardly dissolve
the subject compound (the compound of the formula (0) can be used as "the poor
solvents." They include water; alcohols such as methanol, ethanol and octanol;
i ethers such as diethylether; esters of acetic acids such as ethyl acetate; and
hydrocarbons such as toluene, oyclohexane and hexane.
In case of the crystallization by cooling, the ltoy is to once dissolve the

compound in the good solvenl(s), the mixed solvents) of the good solvents, or the mixed solvent(s) of the good solventCs) and the poor solvent^), and then to cool down the mixture to precipitate the crystals of the suhjoct compound. In ordoi' to once dissolve it, heating ia preferable and the healing temperatero in within the range from 30°C to around the boiling point, of the solvent.
For example, when precipitating the ex -type crystal, it is preferable to dissolve the compound of the formula (I) in a good solvnnt(s) containing at least one kind selected from acetonitrile, dichloromothnno, totrnhydrofurnn, acetone, dimethyl.sulfoxide and chloroform, or in a mixed solvent of ncotonitrilo-water, or in a mixed solvent of acetonitrile-dmiethylformamido; and then con] down the mixture to 0 to 30°C and more preferably 4 to 25"C to crystallize.
In the above, the volume ratio of acetonitrile in the mixed solvent of acetonitrile-water is preferably within the range from 50 to less than 100v/v%, and more preferably from 80 to 9f)v/v%.
When precipitating the y -type crystal, it is preferable to dissolve the compound of the formula (1) in dimethylforniainide and then cool down the mixture tit 0 to 30°C and more preferably 4 to 25"G to crystallize.
In case of the crystallization with a poor solvents), the key is the selection of the good solvents and the poor solvents or the used quail titles thereof. The crystals of the subject compound are precipitated by adding the poor solvent(s) to the good solvent(s). As for the selection of the used HO)vents or the used quantities thereof, the most suitable condition can he Moleelnd by the experiments on solubility.
l:Vor example, when precipitating the rv type crystal, il. is preferable to dissolve the compound of the formula (I) in a good iiolvent(s) and then adding a poor solve nt(s) thereto to crystallize, wherein the combination of the good solvent(s) and the poor solvent(s) is either one of dlmethylsulfoxide-toluene,

dimethylforinamide-diethylether, dimethyl Ibnnainido-toluene,
chloroform ethanel, chloroform-toluene, chloiofornrdiofhylether,
dichloroinethane-diethylefher, totrahydrofuran-water,
tetrahydrofuran-cyclohexane, acetone-water, ncotonitrile-wnter or
dimethylformomidcracetonitrile. In these combinations, the good solvent(s)/the poor solvont(s) is preferably used in 1/20 to 5/1 (volume ratio).
Meanwhile, depending on the combination*! of the good solvents, the above includes addition of the other good solveutCs) to the above good sol von tOO to precipitate the crystals of the subject compound.
For instance, when precipitating the ft-typo crystal, it includes the method comprising the stops of- dissolving the compound of the formula 0) in dimethyl form amide under heating (e.g. 30 to M0"G); adding dropwise I to 20 times volume and more preferably 2 to 8 times volume of neofonlfrilo to the dissolution solution at 0 to 80"C to crystallize.
When precipitating the y -type crystal, it is preferable to dissolve the compound of the formula (I) in a good solvent(s) and then add a poor solvont(s) thereto to crystallize, wherein the combination of the good solvonf(s) and the poor solvent(s) is dimethylformamide-water. In tins case, dlinetbyllbrmamide/wnter is preferably used in 50/1 to 1000/1 (volume ratio).
Besides, when precipitating the £ -type crystal, It is preferable to dissolve the compound of the formula (I) in a good solvont(s) and then add a poor solvent(s) thereto to crystallize, wherein the combination of the good solvent(s) and the poor solvent(s) is either one of diehloromothnne-othanol or dimothylsulfoxide-diethylether. In these combinations, the good solvent(s)/the poor solvent(s) is preferably used in 1/5 to 1/2 (volume ratio).
The crystallization by suspension includes the method which comprises the step.'i of: suspending the compound of the form id a (I) in either one of neotonitrilo,

a mixed HO! vent of acetonitrilo-wafer, ur u mixed solvent of ai^toiufrihrdimethylformamide; and stir tin? mixfuro to crystallize. In case of using the mixed solvent(s), acetonitrile-water or n<'.e(oiulril.irdiniofhylfornKnn:ide it) preferably used in 50/50 to 05/5 (volume ratio).
Whim obtaining the ct 'type crystal, in case of iisini; aeofonilrilirwater (00/10; volume ratio) as a suspending' solvent, it is preferable to suspend the amorphous or the crystals (bore, the crystals mean various crystal forms) of the compound of the formula (I) in the solvent, stir the mixture at 0 to ,'tO"C and isolate the crystals at 0 to 30°C (e.g. at room temperature) by filtering out. At that lime, the subject a -type crystal may bo seeded at 0 to !W°C.
Further, when obtaining the a-type crystal, it is preferable to suspend the amorphous or the crystals (here, the crystals mean various crystal forms) of the compound of the formula (I) at 0 to 40 °C (e.g. at room temperature) in acetonifriledimethylformamide (80/20: volume ratio), stir the mixture at 0 to 40°C and isolate the crystals at 0 to 40°C (e.g. at room temperature) by filtering out. At that time, the subject a -type crystal may be seeded at 0 to 40°C.
On the other hand, when obtaining the 0 -typo crystal, in case of using acetonitrile-water (90/10: volume ratio) as a suspending solvent, if is preferable to suspend the amorphous or the crystals (bore, the crystals mean various crystal forms) of the compound of the formula (I) in the solvent, stir the mixture at 40°C or higher (e.g. 60°C) and isolate the precipitated crystals at 4()UC or higher (e.g. 60°C) by filtering out. In such a case, the maximum temperature is preferably the boiling point of the solvent or that of the mined solvent or lower than if. At that lime, the subject 6 -type crystal may be seeded at 40 to (K)"C.
Besides, when obtaining the 6 -type crystal, if is preferable to suspend the amorphous or the crystals (here, the crystals mean various crystal forms) of the compound of the formula (I) in acetonitriledimethvlfonnamide (80/20: volume

ratio), stir the mixture at 50°C or higher (e.g. (>()UC) anil isolate the precipitated crystals at 50 °C or higher (e.g. 60°C) by Cil te ring on I.. In such a ca.se, the maximum temperature is preferably the boiling point of the solvent or that of the mixed HO I vent or lower than it. At that timet, the Hubject 0 type crystal may be seeded at 50 to 60°C.
The intersection of the saturation solubility curves of the ct -type crystal and the 0 -typo crystal to each solvent system exists at around 30 to 40°C in acetonitrilo-water (90/1()'- volume ratio) and around 40 to 50 °C in acetonilrile (limethylformamido (80/20: volume ratio), At around those temperatures, the mixture of the a -type crystal and the 0 -typo crystal is obtained.
The crystallization by neutralization includes tbo method comprising the steps of dissolving the compound of the formula (I) or a hydrochloride of the compound of the formula (J) in a lower aleobol solution containing hydrogen chloride; and then neutralizing the mixture with a baso(s) to crystallize.
'The bases include inorganic bases such as sodium hydroxide and potassium hydroxide and organic bases such as triethylamino.
The alcohol solutions having 1 to G carbon atoms include methanol, ethanol, propanol, butanol, pentanol and hexanol. Methanol and ethanol are preferable among them.
The crystals of the presont invention are useful in the points that they are
the crystals excellent in "preservation stability" or "moisture resistance" of drug
substances or preparations; and those that can be produced on the industrial
scale.
* Especially, since the a-type crystal is thermodynamically most stable under
room temperature, it can be easily isolated under room temperature and has low hygroscopicity. Since the 0 -type crystal is thermodynamically most stable

under high temperature (50°C or higher), it can bo easily isolated under high temperature (50°C or higher). The 1] -type crystal in also thonnodynainieally stable and the Y"type and / -type crystals are useful in the point of their low hygroscopic] ty.
All of the or-type, 7"-typo, £-type, r/"l,ypn mid 0-typo r.ry»l.nlH con he produced on the industrial scale. Particularly, the (Ytype and 0 -type crystals are preferable in terms of the production on the industrial scale.
The present invention will he further illustrated as follows. The following Examples will illustrate the production examples of the crystals of the present invention. They only explain the present invention and do not particularly limit the invention. Examples
The production of the a-type crystal: Examples I to 2!l The production of the y -type crystal: Examples 2 of the title crystals (yield: 93.8%). (Example 7) The crystallization with a poor solvontbd (the (V typo crystal)
ftOOmg of the compound of the formula (Example 17) The crystallization with a poor solvont(s) (the a -type crystal)
HOOing of the compound of the formula (!) was dissolved in 15mL of acetonitrile. 5mL of water was added to the dissolution solution, and the

precipitated crystals were filtered out and air-dried to obtain the title crystals. (Example 18) The crystallization with a poor Holvont(n) (the iv-type crystal)
4.851, of dimethylformamide was added to VAHH)\\ of the compound of the formula 0) mninly comprising the a type crystals and dissolved by heating at 71°C. 10.4L of acetonitrile WIIH added dropwiso In the dissolution solution at (■>(.» to 75°C. Then, the mixed solution was cooled down to I0°C, the retype crystals were weeded halfway at 54°C and matured at 40"C I'm' '.! hours. Next, the crystallization solution was cooled down to 5"C in 4 hours and stirred overnight. The precipitated crystals were 'filtered out and dried at 00"C under reduced pressure to obtain 173 lg of the title crystals (yield; 811.0% >. (Example 19) The crystallization by suspension (the 0°C under reduced pressure to obtnin 26.G7K <>f U"> title crystals (yield: 83%). (Example 21) The crystallization by suspension (the rv type crystal)
'15.Og of the compound of the formula (I) was added to 112ml., of dimethyl form amide and dissolved at 70°C. Keeping the solution temperature at (!5"C or higher, 445mL of acetonitrile was added dropwiso thereto. After adding it drop wise, the dissolution solution was cooled down to I0"C. Tins crystal obtained by natural crystallization was filtered out and dried at 50 "C under

reduced pressu re to obtain 3(1.8g of the mixture of the l°C. Ig of the 0 type crystals was added thereto and continuously heated at (I0°C for 24 hours. At that time, the partial pullout of the suspension was conducted three times in order to check Uio crystal form (total of 8.71g of the crystals was pulled out). The precipitated crystals were filtered out at 60°C and dried under reduced pressure to obtain 5.0fig of the title crystals. (Example 31) The crystallization by suspension (the 0 -type crystal)
The compound of the formula (I) comprising 2.01 g of the a ■ typo crystals and 1.72g of the 0 -type crystals was added to lOOg of acetonitrilewater (i:i volume ratio) and stirred at 40°C for 1)5 hours. At that time, the partial pullout of the

suspension was conducted twice in order to check the crystal form (total of 1.84g of the crystals were pulled out). The precipitated crystals were filtered out at 40°C and dried under reduced pressure to obtain l.'ICIg of tin* title crystals. (Analytic Example ]) Measurement of powder X-rav diffraction patterns
(1) The measuring method and conditions
Target: Cu Full automatic monochromntor
Voltage: 40kV
Current: 40m V
Slit: divergence 1/2°
scattering 1/2°
light receiving 0.15mm Scan Speed: 2° /min. 2 6 range: 3 to 30°
(2) The measurement results
Figures 1 to 5 show the powder X-ray diffraction patterns of the a-type, y •type, (■: type, ?;-type, and 9 -type crystals, respectively.
Besides, Tables 1 to 5 show the diffraction angles (2 0) el'the main peaks and the strength of each crystals. Table I

cv-|,ypu
10.7 10.8
14.0
14.4 16.0
16.2
17.1
17.2
18.4
18.8
20.6
21.7
23.1
27.8
28.1

Mln.>i|i?lh Hjj'uim
BkmiK
i||ji;i>nK 5Jj;onK
iridium
li|edium
— njmlium mudium
— njeillinii
medium medium
medium

Table Z yty\M

zd il.ii'iiBl-li
7.2 iijmlliiin
~ii.i " hill MlK
10.3 iimillara,.-
ioi ii)udiu)n_
li|iulhiiu
14.5

1^1 j.m!«ll.Lu»_-
18.4*" " iiniillmn_.
v/i" " IUUil.lUl£l_
IflJ U»!').llll)!J_
)QA _ lilVtlL'JILU.
ao.r rnmlluni
I!I. of I'M ten!, Literature I is the compound synthesized in accordance with Example IDG of Patent I .literature ] and that obtained as a hydrochloride was used.

It is clarified from the result of the powder X-ray diffraction in Figure 6 that the compound is amorphous. (Analytic Example 2) Measurement of the infrared absorption spectrum
(1) The measuring method and conditions
The infrared absorption spectrum was measured by FT-lli in accordance with the potassium bromide disk method in the llonornl Tents of the dnpanose Pharmacopoeia.
(2) The measurement results
Figure 7 shows the infrared absorption spectrum til' the 77 -typo > 7'-type>£ type - f/typo in that order. Meanwhile, when the crystal transition occurs in high temperature range, it is thought that the (I iype