EA045975B1 - CRYSTALLINE L-ARGININE SALT (R)-2-(7-(4-CYCLOPENTYL-3-(TRIFLUOROMETHYL)BENZYLOXY)-1,2,3,4-TETRAHYDROCYCLOPENTA[B]INDOL-3-YL)ACETIC ACID (COMPOUND 1 ) FOR USE IN DISORDERS ASSOCIATED WITH THE S1P1 RECEPTOR - Google Patents

Description

Field of technology to which the invention relates

The invention relates, in particular, to a new free-plate crystal form (free-plate crystal habit) or morphology, methods for obtaining a free-plate crystal form and the use of a free-plate crystal form of L-arginine salt (R)-2-(7- (4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid (compound 1) in the treatment of S1P ₁ receptor-related disorders, e.g. lymphocyte-mediated disorders, transplant rejection, autoimmune diseases and disorders, inflammatory diseases and disorders (eg, acute and chronic inflammatory conditions), cancer and conditions whose underlying cause is a violation of vascular integrity, or conditions associated with pathological angiogenesis (which, for example, may occur with inflammation, tumor development and atherosclerosis).

State of the art

Sphingosine 1-phosphate (S1P) receptors 1–5 constitute a family of G protein-coupled receptors with seven transmembrane domains. These receptors, called S1P ₁ - S1P ₅ (originally called endothelial differentiation gene (EDG) receptor-1, -5, -3, -6 and -8, respectively); Chun et. al., Pharmacological Reviews, 54:265-269, 2002) are activated through the binding of sphingosine-1-phosphate, which is produced by sphingosine kinase-catalyzed phosphorylation of sphingosine. S1P ₁ , S1P ₄ and S1P5 receptors activate Gi but not Gq, while S1P2 and S1P ₃ receptors activate both Gi and Gq. The S1P ₃ receptor, unlike the S1P1 receptor, responds to the agonist by increasing the level of intracellular calcium.

U)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid (compound 1) is a potential potent ( EC ₅₀ cAMP, 0.093 nM (human)) and selective (EC ₅₀ β-arrestin, 6.10 nM (S1P1), >10000 nM (S1P2), >10000 nM (S1P3), 147 nM (S1P4) and 24.4 nM (S1P5)) is an orally available drug targeting the S1P1 receptor that is undergoing clinical trials.

In preclinical studies, Compound 1 showed the following estimated lymphocyte reduction IC ₅₀ values for four different species: 0.101 μM (mouse), 0.051 μM (rat), 0.058 μM (dog), and 0.098 μM (monkey). It should be noted that the calculated IC ₅₀ values for lymphocyte reduction indicate their total concentration in plasma, where Compound 1 is highly protein bound (97.8% human, 98.0% rat). Compound 1 has been shown to be effective in a mouse model of experimental autoimmune encephalomyelitis (EAE), mimicking multiple sclerosis. When administered prophylactically, compound 1 prevented the onset and reduced the severity of the disease relative to the diluent until day 25, when administration was stopped. All groups further developed severe disease. Therapeutic administration of Compound 1 was also studied. Treatment began on day 18, when all animals had developed severe disease. Compound 1 was administered from days 18 to 37 and resulted in disease reversal relative to vehicle with efficacy similar to that of fingolimod (i.e., GILENYA®, approved in September 2010 for the treatment of patients with relapsing forms of multiple sclerosis). Similarly, compound 1 was effective in a model of collagen-induced arthritis (CIA). Prophylactic oral administration to female Lewis rats on day 17 after daily oral dosing resulted in a significant reduction in ankle diameter similar to that observed in rats treated with fingolimod or methotrexate. There was also an improvement in the histological parameters of the knees and ankles of CIA rats, indicating that when treated with Compound 1, inhibition of lymphocyte infiltration into the joints suppresses CIA in rodents. Additional information can be found in PCT application serial number PCT/US2009/004265, filed July 22, 2009 (International Publication No. WO2010/011316); PCT application serial number PCT/US2011/000153, filed January 27, 2011 (International Publication No. WO2011/094008); and Buzard: DJ et. al., ACS Med. Chem. Lett. 2014, 5, 1313-1317; the contents of which are included in this description in their entirety by reference.

Clinical trials were conducted using L-arginine salt of Compound 1. A randomized, double-blind, placebo-controlled, Phase 1b clinical trial of L-arginine salt of Compound 1 was conducted at multiple escalating doses in five different dosage groups to evaluate the safety, tolerability, pharmacodynamics and pharmacokinetics. For a total of 21 days, 50 healthy volunteers received L-arginine salt of Compound 1 and 10 healthy volunteers received placebo. In phase 1b clinical trials, administration of L-arginine salt of compound 1 showed a dose-dependent effect of reducing the level of lymphocytes in the blood, with an average decrease relative to the initial level of up to 69%. The lymphocyte count, on average, returned to baseline within one week after completion of the drug.

In response to the growing demand for compounds useful in the treatment of S1P1 receptor-related disorders, L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydro

- 1 045975 cyclopenta[b]indol-3-yl)acetic acid (compound 1) was a new compound of importance.

Accordingly, new and efficient methods for preparing the L-arginine salt of Compound 1 and its associated crystalline morphologies are needed. The authors identified some improvements. These improvements are described in the present invention.

The citation of any reference in this application should not be construed as an admission that such reference is prior art to the present invention.

The essence of the invention

The present invention relates, in particular, to a new free-plate crystal form (free-plate crystal habit) or morphology, methods for preparing the free-plate crystal form and use of the free-plate crystal form of L-arginine salt da-2-(7-(4- cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (compound 1) in the treatment of disorders associated with the S1P ₁ receptor (SiPi receptor- related disorders), such as the disorders described in the present invention.

The discovery of a new crystalline platelet form, referred to herein as the free-plate form, has led to certain advantageous properties such as improved filtration manufacturing processes, formulation stability and/or resistance to degradation, higher degree of crystallinity, lower hygroscopicity (see details in example 8).

Some methods for the preparation of L-arginine salt of L-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indol-3-yl)acetic acid have been described previously; see WO2010/011316 and WO2011/094008.

One aspect of the present invention relates to a method for preparing the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, where this method includes the following steps:

a) hydrolysis of the compound of formula (IIa):

where R ³ represents C1-C6 alkyl; in the presence of a hydrolyzing mixture containing lipase and a solvent for the hydrolysis step to obtain da-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid;

b) adding L-arginine to a salt-forming mixture containing ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, water-miscible antisolvent and H ₂ O, to obtain the first mixture containing L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4 -tetrahydrocyclopenta[b]indol-3-yl)acetic acid;

c) heating the first mixture to the first heating temperature;

d) adding a first additional amount of water-miscible antisolvent to the first mixture to obtain a suspension; And

e) cooling the suspension to obtain the free-plate crystalline form of L-arginine salt L)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indole-3- yl)acetic acid.

Another aspect of the present invention relates to a method for preparing the free-plate crystalline form of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid, where the method includes the following steps:

a) hydrolysis of the compound of formula (IIa):

- 2 045975

where R ³ represents C1-C6; in the presence of a hydrolyzing mixture containing lipase and a solvent of the hydrolysis step to obtain ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indole-3- yl)acetic acid;

b) obtaining a first mixture containing L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acids, water-miscible antisolvent and H2O;

c) heating the first mixture to the first heating temperature to obtain a second mixture;

d) adding a first additional amount of said water-miscible anti-solvent to said second mixture while maintaining said first heating temperature to obtain a suspension;

e) cooling said suspension to a first cooling temperature and then heating it to a second heating temperature;

f) cyclically repeating step e) optionally one or more times, wherein said first cooling temperature of each cycle may be the same or different, and said second heating temperature of each cycle may be the same or different; And

g) cooling said suspension to a final cooling temperature to obtain said free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid.

Another aspect of the present invention relates to a method for preparing the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, where said method includes the following steps:

a) adding L-arginine to a salt-forming mixture containing ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, water-miscible antisolvent and H2O, to obtain the first mixture containing L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl)acetic acid;

b) heating the first mixture to the first heating temperature;

c) adding a first additional amount of water-miscible antisolvent to the first mixture to obtain a suspension; And

d) cooling the suspension to obtain the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indole-3- yl)acetic acid.

Another aspect of the present invention relates to a method for preparing the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, where said method includes the following steps:

a) heating the first mixture containing L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acids, water-miscible antisolvent and water, to the temperature of the first heating to obtain the second mixture;

b) cooling the second mixture to a first cooling temperature, then adding a first additional amount of water-miscible antisolvent to the second mixture while maintaining the first cooling temperature, and then heating to a second heating temperature to obtain a slurry;

c) cyclically repeating step b) optionally once or twice; And

d) cooling the suspension to a second cooling temperature to obtain a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid.

Another aspect of the present invention relates to a method for preparing the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, where said method includes the following steps:

a) obtaining the first mixture containing L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, water-miscible antisolvent and H2O;

- 3 045975

b) heating the first mixture to the first heating temperature to obtain a second mixture;

c) adding a first additional amount of said water-miscible anti-solvent to said second mixture while maintaining said first heating temperature to obtain a suspension;

d) cooling the suspension to the first cooling temperature and then heating to the second heating temperature;

e) cyclically repeating step d) optionally one or more times, where the first cooling temperature of each cycle may be the same or different, and the second heating temperature of each cycle may be the same or different; And

f) cooling said suspension to a final cooling temperature to obtain said free-plate crystalline form of L-arginine salt(s)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid.

Another aspect of the present invention relates to a method for preparing the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, additionally including the step of obtaining the drug free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4 -tetrahydrocyclopenta[b]indol-3-yl)acetic acid to obtain a pharmaceutical composition.

Another aspect of the present invention relates to a method for preparing the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, further comprising the step of mixing the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid with a pharmaceutical excipient to obtain a pharmaceutical composition.

Another aspect of the present invention relates to pharmaceutical compositions obtained in accordance with any of the methods described in the present invention.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid obtained in accordance with any of the methods described in the present invention.

Another aspect of the present invention relates to compositions containing the free-plate crystalline form of L-arginine salt L)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid obtained in accordance with any of the methods described in the present invention.

Another aspect of the present invention relates to pharmaceutical compositions containing a free-plate crystalline form of L-arginine salt(s)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid obtained in accordance with any of the methods described herein, and a pharmaceutical excipient.

Another aspect of the present invention relates to pharmaceutical compositions containing a free-plate crystalline form of L-arginine salt(s)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid (compound 1) in an amount equivalent to a therapeutically effective amount of the free-plate crystalline form of compound 1, wherein the pharmaceutical composition further contains a diluent, a disintegrating agent and a lubricant.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indole-3 -yl)acetic acid.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, the powder X-ray diffraction pattern of which has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2° and 20.5°±0.2 °.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, the differential scanning calorimetry curve of which has an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.0°C to 208.5°C when scanning at a speed of 10°C per minute.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, the dynamic moisture sorption (DSW) plot of which with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the specified free-lamellar crystalline form by about 0.3 wt.% or less at relative

- 4 045975 relative humidity (RH) 90%.

Another aspect of the present invention relates to compositions containing a free-plate crystalline form of L-arginine salt (K)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention.

Another aspect of the present invention relates to pharmaceutical compositions containing a free-plate crystalline form of L-arginine salt (K)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid, which is described in the present invention, and a pharmaceutical excipient.

Another aspect of the present invention relates to methods of treating S1P ₁ receptor-related disorder in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl -3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, which is described in the present invention, composition or pharmaceutical composition, which is described in the present invention.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the manufacture of a medicament for the treatment of S1P ₁ -receptor-related disorder.

Another aspect of the present invention relates to the use of a composition containing a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid, which is described in the present invention, and an excipient, in the manufacture of a medicament for administration in the treatment of S1P ₁ receptor-related disorder.

Another aspect of the present invention relates to the use of a composition containing a free-plate crystalline form of L-arginine salt (L)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid, which is described in the present invention, and an excipient, for preparing a drug for administration in the treatment of S1P ₁ receptor-related disorder.

Another aspect of the present invention relates to the use of a composition containing a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid, which is described in the present invention, and an excipient, for compounding a drug for administration in the treatment of S1P ₁ receptor-related disorder.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating a human or animal with a therapy.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating S1P ₁ receptor-related disorder.

In some embodiments of the present invention, the S1P ₁ receptor-related disorder is selected from primary biliary cirrhosis, psoriasis, psoriatic arthritis, rheumatoid arthritis, Crohn's disease, multiple sclerosis, inflammatory bowel disease, systemic lupus erythematosus, and ulcerative colitis.

These and other aspects of the present invention disclosed in the specification will now be set forth in more detail.

Brief description of the figures

In fig. Figure 1 shows a microphotograph of a sample of spherulites/radial clusters obtained using scanning electron microscopy (SEM); see example 1 for more information.

In fig. Figure 2 shows a microphotograph of a sample of spherulites/radial clusters obtained using the optical polarization microscopy (OPM) method; see example 1 for more information.

In fig. 3A is a micrograph of the crystal morphology of Example 2 (Lot J2) obtained using scanning electron microscopy (SEM).

In fig. 3B is a micrograph of the crystal morphology of Example 2 (Lot J2).

In fig. 4A is a micrograph of the crystal morphology of Example 2 (Lot J1) obtained using scanning electron microscopy (SEM).

In fig. 4B shows a microphotograph of the crystal morphology of sample example 2 (lot

- 5 045975

J1).

In fig. 5A is a photomicrograph of the platelet crystal morphology sample of Example 3 obtained using optical polarization microscopy (OPM).

In fig. 5B is a micrograph of a sample of the platelet crystal morphology shown in FIG. 5A, as described in Example 3, which was obtained using optical polarization microscopy and further shows the outline of a substantially complete free plate of an elongated hexagonal shape.

In fig. Figure 6 shows a differential scanning calorimetry (DSC) thermogram for lot A1 (plate) of example 3.

In fig. Figure 7 shows a differential scanning calorimetry (DSC) thermogram for lot A2 (plate) of example 3.

In fig. Figure 8 shows a differential scanning calorimetry (DSC) thermogram for lot A3 (plate) of example 3.

In fig. Figure 9 shows a differential scanning calorimetry (DSC) thermogram for lot A4 (plate) of example 3.

In fig. Figure 10 shows a differential scanning calorimetry (DSC) thermogram for lot A5 (plate) of example 3.

In fig. 11 shows a microphotograph of a sample of plate-like crystal morphology of example 4.1 (method 1), which was obtained using the optical polarization microscopy (OPM) method.

In fig. 12A is a photomicrograph of the platelet crystal morphology sample of Example 4.1 (Method 1), which was obtained using optical polarization microscopy (OPM).

In fig. 12B shows a differential scanning calorimetry (DSC) thermogram of the sample of Example 4.1, method 1 (wafers).

In fig. Figure 13 shows an overlay of PXRD XRD patterns of Lot A5 and the sample of Example 4.2 (Method 2), which shows that the sample has the same crystalline phase as Lot A5 (wafers).

In fig. Figure 14 shows a microphotograph of sample example 4.2 (method 2), which was obtained using the method of optical polarization microscopy (OPM). The image shows that the sample consists mainly of plates.

In fig. 15 shows a flow diagram for the preparation of the core tablets of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid (compound 1).

In fig. Figure 16 shows the adsorption phase and desorption phase of a dynamic moisture sorption (DMS) analysis of a typical plate morphology lot (i.e. lot A2).

In fig. 17 shows the adsorption/desorption cycle of a typical plate morphology lot (i.e. lot A2).

In fig. 18 shows an overlay of PXRD radiographs of five lots (i.e., Lots A1 to A5, plates) of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3. 4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (compound 1) (see example 3).

In fig. 19 is an overlay of PXRD X-ray diffraction patterns of the L-arginine salt of Compound 1, showing differences in peak intensities between samples of Lot A2 (plates) and Lot H2 (spherulites), which indicates a higher degree of crystallinity of the plates compared to spherulites. The figure also shows the lower background scattering associated with the sample (ie, less amorphous halogen contribution) of Lot A2 (wafer).

In fig. 20 is a micrograph of a crystalline sample of Example 7 (Method 1, Step B; WO2011/094008), which was obtained using optical polarization microscopy (OPM). The image shows that the sample consists of homogeneous particles of radial clusters (spherulites) and a number of small particles.

In fig. Figure 21 shows a block diagram of cleaning (see example 8).

In fig. Figure 22 shows a differential scanning calorimetry (DSC) thermogram of lot A6 (plate) of example 3.

In fig. Figure 23 shows a differential scanning calorimetry (DSC) thermogram of lot A7 (plate) of example 3.

In fig. Figure 24 shows a differential scanning calorimetry (DSC) thermogram of lot A8 (plate) of example 3.

In fig. Figure 25 shows a differential scanning calorimetry (DSC) thermogram of lot A9 (plate) of example 3.

Detailed Description of the Invention Definitions

For the sake of clarity and consistency, the following definitions will be used throughout this patent document.

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The phrase L-arginine salt of compound 1 refers to the L-arginine salt of ^)-2-(1-(-cyclopentyl3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acids:

Connection 1

The phrase which is presented in reference to the figure refers to the crystalline form and/or morphology that is characterized by the graphical data that is presented in the figure. Such data includes, for example, X-ray powder diffraction patterns, differential scanning calorimetry curves, and dynamic moisture sorption plots. One skilled in the art will appreciate that such graphical representations of data may vary slightly, for example, in relative peak intensities and positions due to varying instrument sensitivities and changes in sample concentration and purity, as is well known to those skilled in the art. However, one skilled in the art can simply compare the graphical data presented in the figures of the present invention with graphical data obtained for an unknown crystal form and/or morphology (for an unknown crystal habit) and verify that the two sets of graphical data characterize the same crystal form (or morphology) or two different crystal forms (or morphologies). Thus, it should be borne in mind that the free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid (compound 1) referred to herein as having the graphical data shown in the figure includes any morphologies characterized by said graphical data with minor variations that are known to one skilled in the art to compared with the data presented in the figure.

The term composition refers to a compound or a salt thereof, such as L-arginine salt (R)-2-(7(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, in combination with at least one additional component.

The term preparation as used herein refers to the step(s) of converting a bulk pharmaceutical active ingredient (i.e., FAI) into a drug substance or drug for use by a patient in the treatment of a disease, where the terms treatment and patient have the meanings defined in this description.

The phrase free-plate form or free-plate crystal habit refers to the overall shape of an independent, essentially planar crystal with a length and width that is similar to and substantially greater than the thickness where the plate is not part of a radial cluster such as a spherulite. It should be understood that, due to the fine characterization of crystals, the phrase free-plate form includes whole plates, substantially whole plates, fragments/fragments of plates, and mixtures thereof that are free or substantially free of radial clusters or spherulites.

The terms spherulite and radial cluster refer to a crystalline form (crystal habit) consisting of thin plates or flakes that are clustered radially around a nucleation site that show a cross when viewed with cross-polarized light; see fig. 1 and fig. 2, which shows microphotographs of typical spherulites/radial clusters. Typically, the length of the thin plates forming a radial cluster is less than 10 μm.

The term length in the context of crystalline form refers to the longest edge-to-edge dimension of a particle oriented parallel to the axis of the lens.

The term width in the context of crystalline form refers to the largest dimension of the particle, measured at right angles to the length.

The term patient as used herein refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses or primates, most preferably humans.

The term inert atmosphere, when used herein, refers to an atmosphere substantially free of oxygen. Examples of inert atmospheres include, for example, argon and nitrogen.

The term pharmaceutical composition, when used herein, refers to a composition containing at least one active ingredient, such as compound 1 and its L-arginine salt, such that the composition is suitable for testing for a specific pharmacological effect in a mammal (for example, but without restrictions, human) or for the treatment of a disease or disorder that is described in the present invention. One skilled in the art will be aware of methods suitable for determining whether an active ingredient has

- 7 045975 ent with the desired effective action, based on the requirements of the specialist.

The term treat or treat when used herein includes one or more of the following:

(1) preventing a disease, such as preventing a disease, condition or disorder in a patient who may be susceptible to the disease, condition or disorder but does not yet feel or exhibit the disease pathology or symptoms;

(2) inhibiting a disease, for example inhibiting a disease, condition or disorder in a patient who experiences or exhibits pathology or symptoms of the disease, condition or disorder (ie, stopping further development of the pathology and/or symptoms); and (3) alleviation of a disease, such as alleviating a disease, condition, or disorder in a patient who experiences or exhibits pathology or symptoms of the disease, condition, or disorder (ie, inducing reversal of the pathology and/or symptoms).

Regardless of how much the patient needs treatment, whoever cares for the patient (eg nurse, doctor, physician assistant, nurses, etc. in the case of humans, veterinarian in the case of animals, including non-human mammals) accepts a decision that a patient or animal needs treatment or that treatment will benefit the patient or animal. This decision is made based on a number of factors that are within the control of the person caring for the patient and includes knowledge that the person or animal has or will have a disease, condition or disorder that is treatable with Compound 1 or a pharmaceutically acceptable salt, solvate and hydrate thereof, for example using the L-arginine salt of Compound 1. Accordingly, Compound 1 and/or its pharmaceutically acceptable salts, solvates and hydrates, for example the L-arginine salt of Compound 1, can be used for protection or prophylaxis; or Compound 1 and its pharmaceutically acceptable salts, solvates and hydrates, for example the L-arginine salt of Compound 1, can be used to attenuate, inhibit or alleviate a disease, condition or disorder.

The phrase water-miscible antisolvent, when used herein, refers to a water-soluble solvent in which a compound, such as L-arginine salt (R)2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, poorly soluble.

Chemical group, chemical fragment or radical

The term alkanol refers to a straight or branched carbon radical containing from 2 to 4 carbon atoms and connected to an -OH group. Some embodiments contain 2 to 3 carbon atoms, some other embodiments contain 3 to 4 carbon atoms. Examples of C ₂ -C ₄ alkanol include, but are not limited to, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol and the like.

The term C1- _C6 -alkyl refers to a straight or branched carbon radical containing from 1 to 6 carbon atoms. Some embodiments contain 1 to 5 carbon atoms, some embodiments contain 1 to 4 carbon atoms, some embodiments contain 1 to 3 carbon atoms, and some embodiments contain 1 or 2 carbon atoms. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, nbutyl, sec-butyl, isobutyl, t-butyl, pentyl, isopentyl, t-pentyl, neo-pentyl, 1-methylbutyl [i.e., CH( CH ₃ )CH ₂ CH ₂ CH ₃ ], 2-methylbutyl [i.e. CH ₂ CH(CH ₃ )CH ₂ CH ₃ ], n-hexyl, etc.

It should be understood that certain features of the invention, which are described in the context of individual embodiments for ease of understanding, may also be presented in combination in a single embodiment. Conversely, various features of the invention, which for brevity are described in the context of a single embodiment, may also be presented separately or in any suitable subcombination. In addition, the subcombinations of uses and medical indications listed in the embodiments including the description of such uses and medical indications in the present invention are also specifically covered by the present invention as if each subcombination of uses and medical indications were individually and explicitly described herein.

Some methods of invention

This section describes methods for preparing the crystalline free-plate morphology of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid using enzymatic hydrolysis of the corresponding (R/S) ethyl ester to ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydro cyclopenta[b]indol-3-yl)acetic acid and subsequently preparing the L-arginine salt according to a modified procedure as previously disclosed in WO2011/094008, and isolating the L-arginine salt of Compound 1 in a free-plate crystalline form or morphology.

Accordingly, one aspect of the present invention relates to a method for preparing a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, where said method includes

- 8 045975 includes the following stages:

a) hydrolysis of the compound of formula (IIa):

^F 3 ^C y [yA^COpR ³ (Pa) ^H where R ³ represents C1-C6; in the presence of a hydrolyzing mixture containing lipase and a solvent of the hydrolysis stage, to obtain ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indole-3- yl)acetic acid;

b) adding L-arginine to a salt-forming mixture containing ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, water-miscible antisolvent and H ₂ O, to obtain the first mixture containing L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4 -tetrahydrocyclopenta[b]indol-3-yl)acetic acid;

c) heating the first mixture to the first heating temperature;

d) adding a first additional amount of water-miscible antisolvent to the first mixture to obtain a suspension; And

e) cooling the suspension to obtain the free-plate crystalline form of L-arginine salt L)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indole-3- yl)acetic acid.

In some embodiments, a method for preparing a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indole-3 -yl)acetic acid is carried out in an inert atmosphere. In some embodiments, the method is conducted under an inert atmosphere containing nitrogen or argon. In some embodiments, the method is conducted under an inert atmosphere containing nitrogen.

Step a) - Hydrolysis of the compound of formula (Xa)

In some embodiments of the present invention, the hydrolysis in step a) is carried out under an inert atmosphere. In some embodiments, the hydrolysis in step a) is carried out under an inert atmosphere containing nitrogen or argon. In some embodiments, the hydrolysis in step a) is carried out under an inert atmosphere containing nitrogen.

In some embodiments of the present invention, R ³ is methyl or ethyl. In some embodiments, R ³ is ethyl.

In some embodiments, the lipase is Candida Antarctica lipase B. In some embodiments, the lipase is immobilized Candida antarctica lipase B.

In some embodiments of the present invention, the hydrolysis step solvent contains dimethylformamide (DMF), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), or acetonitrile. In some embodiments, the hydrolysis step solvent contains acetonitrile.

In some embodiments of the present invention, a compound of formula (IIa) is

the lipase is immobilized Candida antarctica lipase B; and the hydrolysis step solvent contains acetonitrile.

In some embodiments of the present invention, the hydrolysis in step a) is carried out in the presence of a phosphate buffer. In some embodiments, the hydrolysis in step a) is carried out in the presence of a phosphate buffer at a pH of from about 6.0 to about 9.0. In some embodiments, the hydrolysis in step a) is carried out in the presence of a phosphate buffer at a pH of from about 7.0 to about 8.5. In some embodiments, the hydrolysis in step a) is carried out in the presence of a phosphate buffer at a pH of from about 7.3 to about 8.3. In some embodiments, the hydrolysis in step a) is carried out in the presence of a phosphate buffer at a pH of from about 7.6 to about 8.0. In some embodiments, the hydrolysis in step a) is carried out in the presence of a phosphate buffer at a pH of about 7.8. In some embodiments, the phosphate buffer is a sodium phosphate buffer. In some embodiments, the phosphate buffer

- 9 045975 is a potassium phosphate buffer.

In some embodiments of the present invention, the hydrolysis in step a) is carried out at a temperature of from about 0°C to about 75°C. In some embodiments, the hydrolysis in step a) is carried out at a temperature of from about 20°C to about 65°C. In some embodiments, the hydrolysis in step a) is carried out at a temperature of from about 30°C to about 55°C. In some embodiments, the hydrolysis in step a) is carried out at a temperature of from about 35°C to about 45°C.

In some embodiments of the present invention, the hydrolysis in step a) further includes the step of isolating N)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indole-3 - yl)acetic acid.

In some embodiments of the present invention, after isolation of the enantiomeric excess of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3 yl)acetic acid is approximately 95% or more. In some embodiments, after isolation, the enantiomeric excess of A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol-3-yl)acetic acid is approximately 98 % or more. In some embodiments, after isolation, the enantiomeric excess of A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid is approximately 99% or more.

Step b) - Adding L-arginine to the salt-forming mixture

In some embodiments of the present invention, the addition in step b) is carried out under an inert atmosphere. In some embodiments, the addition in step b) is carried out under an inert atmosphere containing argon or nitrogen. In some embodiments, the addition in step b) is carried out under an inert atmosphere containing nitrogen.

In some embodiments of the present invention, the water-miscible antisolvent in step b) includes a solvent selected from the group consisting of acetonitrile, acetone, tetrahydrofuran and a C ₂ -C ₄ alkanol. In some embodiments, the water-miscible antisolvent in step b) includes a solvent selected from the group consisting of acetonitrile, acetone, tetrahydrofuran, ethanol, 1-propanol, 2-propanol and 1-butanol. In some embodiments, the water-miscible antisolvent in step b) includes 2-propanol.

In some embodiments, the molar ratio of A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L-arginine is about 1.0:0.95 to about 1.0:1.2. In some embodiments, the molar ratio of N)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]yne dol-3-yl)acetic acid and L -arginine is from about 1.0:1.0 to about 1.0:1.2. In some embodiments, the molar ratio of A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indol-3-yl)acetic acid and L- arginine is approximately 1.0:1.0.

In some embodiments of the present invention, the temperature of the salt-forming mixture before adding L-arginine is from about 15°C to about 83°C. In some embodiments, the temperature of the salt-forming mixture before adding L-arginine is from about 18°C to about 80°C. In some embodiments, the temperature of the salt-forming mixture before adding L-arginine is from about 20°C to about 40°C.

Another aspect of the present invention allows L-arginine to be added to the salt-forming mixture as a solid.

In some embodiments of the present invention, the addition of L-arginine in step b) is accomplished by adding L-arginine as a solid to the salt-forming mixture.

In some embodiments of the present invention, the addition of L-arginine in step b) is accomplished by adding substantially all of the L-arginine as a solid to the salt-forming mixture. The phrase essentially all at once or all at once refers to the addition of the entire amount of L-arginine to the salt-forming mixture at one time, with the only limitation being the equipment used.

In some embodiments of the present invention, the addition of L-arginine in step b) is accomplished by adding L-arginine as a solid to the salt-forming mixture over a period of 30 minutes. In some embodiments, the addition of L-arginine in step b) is accomplished by adding L-arginine as a solid to the salt-forming mixture over a period of 1 hour. In some embodiments, the addition of L-arginine in step b) is accomplished by adding L-arginine as a solid to the salt-forming mixture over a period of 2 hours.

In some embodiments of the present invention, the salt-forming mixture comprises (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid nic acid, 2-propanol and water in a weight ratio of about 1.00:4.98:0.94 to about 1.00:7.46:1.40. In some embodiments, the salt-forming mixture comprises (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid

- 10 045975 slot, 2-propanol and water in a mass ratio of from about 1.00:5.29:0.99 to about 1.00:7.15:1.35. In some embodiments, the salt-forming mixture comprises (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, 2- propanol and water in a mass ratio of from about 1.00:5.60:1.05 to about 1.00:6.84:1.29. In some embodiments, the salt-forming mixture comprises (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, 2- propanol and water in a weight ratio of from about 1.00:5.72:1.08 to about 1.00:6.72:1.26. In some embodiments, the salt-forming mixture includes ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid, 2- propanol and water in a weight ratio of from about 1.00:5.85:1.10 to about 1.00:6.59:1.24. In some embodiments of the present invention, the salt-forming mixture includes ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid , 2-propanol and water in a weight ratio of about 1.00:5.97:1.12 to about 1.00:6.47:1.22. In some embodiments of the present invention, the salt-forming mixture includes (C)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid , 2-propanol and water in a weight ratio of about 1.00:6.10:1.15 to about 1.00:6.34:1.19. In some embodiments of the present invention, the salt-forming mixture comprises (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, 2-propanol and water in a mass ratio of approximately 1.00:6.22:1.17.

In some embodiments of the present invention, the temperature of the salt-forming mixture upon addition of L-arginine is from about 15°C to about 83°C. In some embodiments, the temperature of the salt-forming mixture upon addition of L-arginine is from about 18°C to about 80°C. In some embodiments, the temperature of the salt-forming mixture upon addition of L-arginine is from 20°C to about 40°C.

Another aspect of the present invention allows L-arginine to be added to the salt-forming mixture in the form of an aqueous solution.

In some embodiments of the present invention, the addition of L-arginine in step b) is accomplished by adding L-arginine in the form of an aqueous solution to the salt-forming mixture.

In some embodiments, the addition of L-arginine in step b) is accomplished by adding L-arginine as a solution to the salt-forming mixture, wherein the L-arginine solution is an aqueous solution of L-arginine at a concentration of from about 2.1 M to about 2.3 M and temperatures from about 50°C to about 75°C. In some embodiments, the addition of L-arginine in step b) is accomplished by adding L-arginine as a solution to the salt-forming mixture, wherein the L-arginine solution is an aqueous solution of L-arginine at a concentration of about 2.26 M to about 2.28 M at a temperature from about 55°C to about 65°C.

In some embodiments of the present invention, after adding L-arginine as an aqueous solution in step b), the weight ratio is ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid, 2-propanol and H2O is from about 1.00:4.98:0.94 to about 1.00:7.46:1.40. In some embodiments, after adding L-arginine as an aqueous solution in step b), the weight ratio of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] ]indol-3-yl)acetic acid, 2-propanol and H2O is from about 1.00:5.29:0.99 to about 1.00:7.15:1.35. In some embodiments, after adding L-arginine as an aqueous solution in step b), the weight ratio of (D)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid, 2-propanol and H2O is from about 1.00:5.60:1.05 to about 1.00:6.84:1.29. In some embodiments, after adding L-arginine as an aqueous solution in step b), the weight ratio of (D)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[B] ]indol-3-yl)acetic acid, 2-propanol and H2O is from about 1.00:5.72:1.08 to about 1.00:6.72:1.26. In some embodiments, following the addition of L-arginine as an aqueous solution in step b), the weight ratio is ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid, 2-propanol and H2O is from about 1.00:5.85:1.10 to about 1.00:6.59:1.24. In some embodiments, after adding L-arginine as an aqueous solution in step b), the weight ratio of (R)2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid, 2-propanol and H2O is from about 1.00:5.97:1.12 to about 1.00:6.47:1.22. In some embodiments, after adding L-arginine as an aqueous solution in step b), the weight ratio of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[ b]indol-3-yl)acetic acid, 2-propanol and H2O is from about 1.00:6.10:1.15 to about 1.00:6.34:1.19. In some embodiments, following the addition of L-arginine as an aqueous solution in step b), the weight ratio is ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid, 2-propanol

- 11 045975 and H2O is from about 1.00:6.22:1.17.

In some embodiments of the present invention, step b) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, step b) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step b) is carried out at a mixing speed of approximately 150 rpm.

Stage c) - Heating the first mixture to the first heating temperature

In some embodiments of the present invention, the first heating temperature is from about 20°C to about 83°C. In some embodiments, the first heating temperature is from about 20°C to about 55°C. In some embodiments, the first heating temperature is from about 25°C to about 45°C. In some embodiments, the first heating temperature is from about 50°C to about 83°C. In some embodiments, the first heating temperature is from about 60°C to about 80°C. In some embodiments, the first heating temperature is from about 70°C to about 80°C.

In some embodiments of the present invention, the first mixture is a substantially homogeneous solution.

In some embodiments, step c) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, step c) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step c) is carried out at a mixing speed of approximately 150 rpm.

Step d) - Adding a first additional amount of water-miscible antisolvent to the first mixture to obtain a suspension

In some embodiments, the water-miscible antisolvent in step b) includes a solvent selected from the group consisting of acetonitrile, acetone, tetrahydrofuran and C.'2-Sgalcanol. In some embodiments, the water-miscible antisolvent in step b) includes a solvent selected from the group consisting of acetonitrile, acetone, tetrahydrofuran, ethanol, 1-propanol, 2-propanol and 1-butanol. In some embodiments, the water-miscible antisolvent in step b) includes 2-propanol.

In some embodiments of the present invention, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:5.46 to about 1.00:8.20. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid and the first additional amount of 2-propanol is from about 1.00:5.81 to about 1.00:7.86. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid and the first additional amount of 2-propanol is from about 1.00:6.15 to about 1.00:7.51. In some embodiments of the present invention, the weight ratio of L-arginine salt (D)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid and the first additional amount of 2propanol is from about 1.00:6.28 to about 1.00:7.38. In some embodiments of the present invention, the weight ratio of L-arginine salt (D)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid and the first additional amount of 2-propanol is from about 1.00:6.42 to about 1.00:7.24. In some embodiments of the present invention, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3 yl )acetic acid and the first additional amount of 2-propanol is from about 1.00:6.56 to about 1.00:7.10. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid and the first additional amount of 2-propanol is from about 1.00:6.69 to about 1.00:6.97. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid and the first additional amount of 2-propanol is approximately 1.00:6.83.

In some embodiments of the present invention, the addition of the first additional amount of 2-propanol to the first mixture is carried out at a rate such that the addition is completed within about 5.00 hours to about 10.00 hours. In some embodiments, the addition of the first additional amount of 2-propanol to the first mixture is carried out at a rate such that the addition is completed within about 6.75 hours to about 8.25 hours. In some embodiments, adding a first additional amount of 2-propanol to

- 12 045975 the first mixture is carried out at a rate at which the addition is completed within from about 6.90 hours to about 8.10 hours. In some embodiments, the addition of the first additional amount of 2-propanol to the first mixture is carried out at a rate such that the addition is completed within about 7.05 hours to about 7.95 hours. In some embodiments, the addition of the first additional amount of 2-propanol to the first mixture is carried out at a rate such that the addition is completed within about 7.20 hours to about 7.80 hours. In some embodiments, the addition of the first additional amount of 2-propanol to the first mixture is carried out at a rate such that the addition is completed within about 7.35 hours to about 7.65 hours. In some embodiments, the addition of the first additional amount of 2-propanol to the first mixture is carried out at a rate such that the addition is completed within about 7.50 hours.

In some embodiments of the present invention, the first heating temperature is maintained during the addition of the first additional amount of 2-propanol to the first mixture.

In some embodiments of the present invention, after adding the first additional amount of 2-propanol to the first mixture in step c), the method further includes preparing a second mixture while maintaining the first heating temperature, where the second mixture is formed before obtaining a suspension.

In some embodiments of the present invention, the method further includes cooling the second mixture to a first cooling temperature followed by heating to a second heating temperature to obtain a slurry.

In some embodiments of the present invention, cooling the second mixture to the first cooling temperature is carried out at a rate of from about 8.80°C/hour to about 14.40°C/hour. In some embodiments, cooling the second mixture to the first cooling temperature is carried out at a rate of from about 9.35°C/hour to about 13.80°Ο ^γ 43Ο. In some embodiments, cooling the second mixture to the first cooling temperature is carried out at a rate of from about 9.90°C/hour to about 13.20°C/hour. In some embodiments, cooling the second mixture to the first cooling temperature is carried out at a rate of from about 10.45°C/hour to about 12.60°C/hour. In some embodiments, cooling the second mixture to the first cooling temperature is carried out at a rate of from about 10°C/hour to about 12°C/hour.

In some embodiments of the present invention, the first cooling temperature is from about 15°C to about 40°C. In some embodiments, the first cooling temperature is from about 20°C to about 30°C. In some embodiments, the first cooling temperature is from about 22°C to about 24°C.

In some embodiments of the present invention, the first cooling temperature is maintained for at least 1 hour before heating to the second heating temperature.

In some embodiments of the present invention, the second heating temperature is from about 65°C to about 83°C. In some embodiments, the second heating temperature is from about 70°C to about 80°C. In some embodiments, the second heating temperature is from about 70°C to about 75°C.

In some embodiments of the present invention, after heating to the second heating temperature, the second heating temperature is maintained for at least 30 minutes.

In some embodiments of the present invention, step d) is carried out at a mixing speed of from about 100 revolutions per minute to about 200 revolutions per minute. In some embodiments, step d) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step d) is carried out at a mixing speed of about 150 rpm.

Step e) - Cooling of the suspension to obtain the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b ]indol-3-yl)acetic acid

In some embodiments of the present invention, the cooling of the suspension in step e) is carried out at a rate of from about 8.8°C/hour to about 14.4°C/hour. In some embodiments, the cooling of the slurry in step e) is carried out at a rate of from about 9.4°C/hour to about 13.8°C/hour. In some embodiments, the cooling of the slurry in step e) is carried out at a rate of from about 9.9°C/hour to about 13.2°C/hour. In some embodiments, the cooling of the slurry in step e) is carried out at a rate of from about 10.5°C/hour to about 12.6°C/hour. In some embodiments, the cooling of the suspension in step e) is carried out at a rate of from about 10°C/hour to about 12°C/hour.

In some embodiments of the present invention, after cooling in step e), the temperature of the suspension is from about 15°C to about 40°C. In some embodiments, after cooling in step e), the temperature of the slurry is from about 20° C. to

- 13 045975 approximately 30°C. In some embodiments, after cooling in step e), the temperature of the slurry is from about 22°C to about 24°C.

In some embodiments, step e) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, step e) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step e) is carried out at a mixing speed of approximately 150 rpm.

In some embodiments of the present invention, the method further includes the step of isolating the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid.

In some embodiments of the present invention, the isolation step comprises filtering the free-plate crystalline form of L-arginine salt (B)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid from the suspension. In some embodiments, the isolation step includes filtering the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid from the suspension and drying the free-plate crystalline form of L-arginine salt under reduced pressure.

In some embodiments of the present invention, after filtration, the onset temperature of the phase transition of the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [B]indol-3-yl)acetic acid ranges from 205.0°C to 208.5°C as determined by differential scanning calorimetry scanning at 10°C/min. In some embodiments, after filtration, the onset temperature of the phase transition of the free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta ]indol-3-yl)acetic acid is from 205.5°C to 208.5°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtration, the onset temperature of the phase transition of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b ]indol-3-yl)acetic acid is from 206.5°C to 208.5°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtration, the onset temperature of the phase transition of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] ]indol-3-yl)acetic acid is from 207.0°C to 208.1°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtering the free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent) onto the differential scanning calorimetry curve [ b]indol-3-yl)acetic acid obtained by scanning at a rate of 10°C/min has an endotherm as substantially represented in any of Figures 6 through 10. In some embodiments, after filtration on the differential scanning calorimetry curve free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, obtained by scanning at a rate of 10°C/min, there is an endotherm as substantially represented in any of Figures 6 to 10 and 22 to 25. In some embodiments, after filtration, a plot of dynamic moisture sorption (DMS) of the free-plate crystal form L -arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid with an adsorption phase of 30 %RH to 90%RH shows an increase in the mass of the free-plate crystalline form of approximately 0.3% by mass or less at 90% RH. In some embodiments, after filtration, the dynamic adsorption plot of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid with an adsorption phase from 30% RH to 90% RH shows an increase in the weight of the free-lamellar crystalline form of about 0.2% by weight or less at 90% RH. The DSW characteristics provided in the present invention may also vary by plus/minus 0.15% change in mass (ie, ±0.15% change in mass).

In some embodiments of the present invention, after filtration, a plot of the total impurities in the free-plate crystalline form of L-arginine salt (R)-2 -(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid obtained by achiral HPLC shows that the total impurity content is 0.5 area% or less. In some embodiments, after filtration, a plot of the total impurities in the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid

- 14 045975 u, obtained by achiral HPLC, shows that the total impurity content is 0.3 area% or less. In some embodiments, after filtration, a plot of the total impurities in the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid obtained by achiral HPLC shows that the total impurity content is 0.2 area% or less.

In some embodiments of the present invention, the method further includes the step of preparing a free-plate crystalline form of L-arginine salt (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid to obtain a pharmaceutical composition.

In some embodiments of the present invention, the method further includes the step of mixing the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid with a pharmaceutical excipient to obtain a pharmaceutical composition.

Another aspect of the present invention relates to a method for preparing the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, which includes the following steps:

a) hydrolysis of the compound of formula (IIa):

where R ³ represents ^-^-alkyl; in the presence of a hydrolyzing mixture containing lipase and a solvent of the hydrolysis step to obtain ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indole-3- yl)acetic acid;

b) adding L-arginine to a salt-forming mixture containing ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, 2-propanol and H2O, to obtain the first mixture containing L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid;

c) heating the first mixture to the first heating temperature;

d) adding a first additional amount of 2-propanol to the first mixture to obtain a suspension; And

e) cooling the suspension to obtain the free-plate crystalline form of L-arginine salt L)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indole-3- yl)acetic acid. Further description of embodiments of these methods and the associated steps a) to e) can be found herein, see above, for example steps a) to e), respectively.

In addition, this invention describes methods for preparing the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indole -3-yl)acetic acid starting from ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acids.

Accordingly, one aspect of the present invention relates to a method for preparing a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, where said method includes the following steps:

a) adding L-arginine to a salt-forming mixture containing ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, water-miscible antisolvent and H ₂ O, to obtain the first mixture containing L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4 -tetrahydrocyclopenta[b]indol-3-yl)acetic acid;

b) heating the first mixture to a first temperature;

c) adding a first additional amount of water-miscible antisolvent to the first mixture to obtain a suspension; And

d) cooling the suspension to obtain the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indole-3- yl)acetic acid. Further description of embodiments of this method which are associated with steps a) to d) can be found in the present description, see above, for example steps a) to e), respectively.

Another aspect of the present invention relates to a method for producing free-lamellar

- 15 045975 crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, where said method includes the following steps:

a) adding L-arginine to a salt-forming mixture containing ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, 2-propanol and H2O, to obtain the first mixture containing L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid;

b) heating the first mixture to a first temperature;

c) adding a first additional amount of 2-propanol to the first mixture to obtain a suspension; And

d) cooling the suspension to obtain a free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indole-3 -yl)acetic acid. Further description of embodiments of this method which are associated with steps a) to d) can be found in the present description, see above, for example steps a) to e), respectively.

Another aspect of the present invention relates to a method for preparing the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, where said method includes the following steps:

a) hydrolysis of the compound of formula (IIa):

where R ³ represents AA-alkyl; in the presence of a hydrolyzing mixture containing lipase and a solvent of the hydrolysis step to obtain ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indole-3- yl)acetic acid;

b) obtaining a first mixture containing L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid, water-miscible antisolvent and H2O;

c) heating the first mixture to the first heating temperature to obtain a second mixture;

d) adding a first additional amount of water-miscible antisolvent to the second mixture while maintaining the first heating temperature to obtain a suspension;

e) cooling the suspension to the first cooling temperature followed by heating to the second heating temperature;

f) cycling step e) optionally one or more times, where the first cooling temperature of each cycle may be the same or different, and the second heating temperature of each cycle may be the same or different; And

g) cooling the suspension to the final cooling temperature to obtain a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid.

In some embodiments of the present invention, a method for preparing a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3 -yl)acetic acid is carried out in an inert atmosphere. In some embodiments, the method is carried out in an inert atmosphere containing nitrogen or argon. In some embodiments, the method is carried out in an atmosphere containing nitrogen.

Step a) - Hydrolysis of the compound of formula (IIa)

In some embodiments of the present invention, the hydrolysis in step a) is carried out under an inert atmosphere. In some embodiments, the hydrolysis in step a) is carried out under an inert atmosphere containing nitrogen or argon. In some embodiments, the hydrolysis in step a) is carried out under an inert atmosphere containing nitrogen.

In some embodiments of the present invention, R ³ is methyl or ethyl. In some embodiments, R ³ is ethyl.

In some embodiments, the lipase is Candida antarctica lipase B. In some embodiments, the lipase is immobilized Candida antarctica lipase B.

In some embodiments of the present invention, the hydrolysis step solvent includes dimethylformamide (DMF), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), or acetonitrile. In some embodiments, the hydrolysis step solvent

- 16 045975 includes acetonitrile.

In some embodiments of the present invention, a compound of formula (IIa) is

the lipase is immobilized Candida antarctica lipase B; and the hydrolysis step solvent includes acetonitrile.

In some embodiments of the present invention, the hydrolysis is carried out in the presence of a phosphate buffer. In some embodiments, the hydrolysis is carried out in the presence of a phosphate buffer at a pH of from about 6.0 to about 9.0. In some embodiments, the hydrolysis is carried out in the presence of a phosphate buffer at a pH of from about 6.9 to about 8.1. In some embodiments, the hydrolysis is carried out in the presence of a phosphate buffer at a pH of from about 7.0 to about 8.5. In some embodiments, the hydrolysis is carried out in the presence of a phosphate buffer at a pH of from about 7.3 to about 8.3. In some embodiments, the hydrolysis is carried out in the presence of a phosphate buffer at a pH of from about 7.6 to about 8.0. In some embodiments, the hydrolysis is carried out in the presence of a phosphate buffer at a pH of about 7.8. In some embodiments, the phosphate buffer is a sodium phosphate buffer. In some embodiments, the phosphate buffer is a potassium phosphate buffer.

In some embodiments of the present invention, the hydrolysis is carried out at a temperature of from about 0°C to about 75°C. In some embodiments, the hydrolysis is carried out at a temperature of from about 20°C to about 65°C. In some embodiments, the hydrolysis is carried out at a temperature of from about 30°C to about 55°C. In some embodiments, the hydrolysis is carried out at a temperature of from about 35°C to about 45°C.

In some embodiments of the present invention, the hydrolysis further includes the step of isolating ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl) acetic acid.

In some embodiments of the present invention, after isolating (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, its enantiomeric the excess is approximately 95% or more. In some embodiments, after isolation of ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol-3-yl)acetic acid, its enantiomeric excess is approximately 98% or more. In some embodiments, after isolation of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, its enantiomeric excess is approximately 99% or more.

In some embodiments of the present invention, after hydrolysis in step a), (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl ) acetic acid is not isolated.

In some embodiments of the present invention, after hydrolysis in step a), (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl ) acetic acid is present in an amount of at least 40%, as determined by HPLC. In some embodiments, after hydrolysis in step a) ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid present in an amount of at least 45% as determined by HPLC.

Stage b) - Preparation of the first mixture

In some embodiments of the present invention, the preparation of the first mixture in step b) is carried out under an inert atmosphere. In some embodiments, the preparation of the first mixture in step b) is carried out under an inert atmosphere containing argon or nitrogen. In some embodiments, the preparation of the first mixture in step b) is carried out under an inert atmosphere containing nitrogen.

It is clear that the preparation of the first mixture containing L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid, water-miscible antisolvent and H ₂ O, can be performed in any manner conventional to one skilled in the art.

For example, preparing the first mixture may involve preparing the L-arginine salt by reacting L-arginine with ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b ]indol-3-yl)acetic acid; and mixing L-arginine, water-miscible antisolvent and H2O.

- 17 045975

As another example, the preparation of the first mixture may include the preparation of a mixture of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid, water-miscible antisolvent and H ₂ O; and reacting L-arginine with (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid to obtain the first mixture .

As another example, the preparation of the first mixture may include the preparation of a mixture of (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3 - yl)acetic acid and water-miscible antisolvent; and the interaction of L-arginine in the form of an aqueous suspension or solution with A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3- yl)acetic acid to obtain the first mixture.

As another example, the preparation of the first mixture may include the preparation of a mixture of (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3 - yl)acetic acid, water-miscible antisolvent and L-arginine; and adding H2O to obtain the first mixture.

Accordingly, in some embodiments of the present invention, preparation of the first mixture in step b) includes the step of adding to the salt-forming mixture containing (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4 -tetrahydrocyclopenta[b]indol-3-yl)acetic acid and water-miscible antisolvent, L-arginine and H2O, together or separately in any order, to obtain the first mixture.

In some embodiments of the present invention, the addition of L-arginine and H2O is carried out under an inert atmosphere. In some embodiments, the addition of L-arginine and H2O is carried out under an inert atmosphere containing argon or nitrogen. In some embodiments, the addition of L-arginine and H2O is carried out under an inert atmosphere containing nitrogen.

In some embodiments of the present invention, the water-miscible antisolvent includes a solvent selected from the group consisting of acetonitrile, acetone, tetrahydrofuran and N-alkanol. In some embodiments, the water-miscible antisolvent includes a solvent selected from the group consisting of acetonitrile, acetone, tetrahydrofuran, ethanol, 1-propanol, 2-propanol, and 1-butanol. In some embodiments, the water-miscible antisolvent includes 2-propanol.

In some embodiments of the present invention, before adding L-arginine and H2O, the salt-forming mixture contains A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indole -3-yl)acetic acid and 2-propanol in a weight ratio of from about 1.0:3.0 to about 1.0:11.0.

In some embodiments of the present invention, before adding L-arginine and H2O, the salt-forming mixture contains A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indole-3 -yl)acetic acid and 2-propanol in a mass ratio of about 1.0:4.0 to 1.0:10.0. In some embodiments, prior to the addition of L-arginine and H2O, the salt-forming mixture contains A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol-3-yl )acetic acid and 2-propanol in a mass ratio of from about 1.0:5.0 to about 1.0:9.0. In some embodiments, prior to the addition of L-arginine and H2O, the salt-forming mixture contains A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol-3-yl )acetic acid and 2-propanol in a weight ratio of from about 1.0:6.0 to about 1.0:8.0.

In some embodiments of the present invention, the molar ratio of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L -arginine is from about 1.0:0.8 to about 1.0:1.2.

In some embodiments of the present invention, the molar ratio of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L -arginine is from about 1.0:0.9 to about 1.0:1.2. In some embodiments, the molar ratio of N)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L -arginine is from about 1.0:1.0 to about 1.0:1.2. In some embodiments, the molar ratio of A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L-arginine ranges from about 1.0:0.93 to about 1.0:1.01. In some embodiments, the molar ratio of N)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]yne dol-3-yl)acetic acid and L -arginine is from about 1.0:0.93 to about 1.0:0.97. In some embodiments, the molar ratio of A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L- arginine is approximately 1.0:1.0. In some embodiments, the molar ratio of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L-arginine is approximately 1.0:0.95.

In some embodiments of the present invention, the weight ratio of L-arginine

- 18 045975 and H2O is from about 1.0:0.4 to about 1.0:2.3. In some embodiments, the weight ratio of L-arginine to H2O is from about 1.0:1.6 to about 1.0:2.1. In some embodiments, the weight ratio of L-arginine to H2O is from about 1.0:0.8 to about 1.0:1.9. In some embodiments, the weight ratio of L-arginine to H2O is from about 1.0:1.0 to about 1.0:1.7. In some embodiments, the weight ratio of L-arginine to H2O is from about 1.0:1.1 to about 1.0:1.6. In some embodiments, the weight ratio of L-arginine to H2O is from about 1.0:1.2 to about 1.0:1.5.

In some embodiments of the present invention, the temperature of the salt-forming mixture before adding L-arginine is from about 15°C to about 83°C. In some embodiments, the temperature of the salt-forming mixture before adding L-arginine is from about 18°C to about 80°C. In some embodiments, the temperature of the salt-forming mixture before adding L-arginine is from about 20°C to about 40°C. In some embodiments, the temperature of the salt-forming mixture before adding L-arginine is from about 18°C to about 30°C.

In some embodiments of the present invention, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine in the form of an aqueous suspension.

In some embodiments of the present invention, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine as a solid.

In some embodiments, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine as a solid substantially all at once. The phrase essentially all at once or all at once refers to the addition of the entire amount of L-arginine to the salt-forming mixture at one time, with the only limitation being the equipment used.

In some embodiments of the present invention, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine as a solid to the salt-forming mixture over a period of about 30 minutes. In some embodiments, the addition of L-Arginine to the salt-forming mixture is accomplished by adding L-arginine as a solid to the salt-forming mixture over a period of about 1 hour. In some embodiments, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine as a solid to the salt-forming mixture over a period of about 2 hours.

In some embodiments of the present invention, the salt-forming mixture contains (R)-2-(7-(4-cyclopentyl-3-(prifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3 yl) acetic acid, 2-propanol and water in a weight ratio of from about 1.0:3.0:0.05 to about 1.0:11.0:1.0. In some embodiments, the salt-forming mixture contains (R)-2-(7-(4cyclopentyl-3-(prifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid, 2-propanol and water in a mass ratio of from about 1.0:4.0:0.1 to about 1.0:10.0:0.9. In some embodiments, the salt-forming mixture contains 4)-2-(7-(4-cyclopentyl-3(prifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, 2-propanol and water in a mass ratio of from about 1.0:5.0:0.15 to about 1.0:9.0:0.8. In some embodiments of the present invention, the salt-forming mixture contains 4)-2-(7-(4-cyclopentyl-3(prifluoromethyl)benzyloxy)-1,2,3,4-tetracyclopenta[b]indol-3-yl)acetic acid, 2-propanol and water in a mass ratio of from about 1.0:6.0:0.25 to about 1.0:8.0:0.7.

In some embodiments of the present invention, the salt-forming mixture contains (R)-2-(7-(4-cyclopentyl-3-(prifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3 yl) acetic acid, 2-propanol and water in a mass ratio of from about 1.00:4.98:0.94 to about 1.00:7.46:1.40, from about 1.00:5.29:0, 99 to about 1.00:7.15:1.35, from about 1.00:5.60:1.05 to about 1.00:6.84:1.29, from about 1.00:5, 72:1.08 to about 1.00:6.72:1.26, from about 1.00:5.85:1.10 to about 1.00:6.59:1.24, from about 1, 00:5.97:1.12 to about 1.00:6.47:1.22, from about 1.00:6.10:1.15 to about 1.00:6.34:1.19, or approximately 1.00:6.22:1.17.

In some embodiments of the present invention, the temperature of the salt-forming mixture at the time of addition of L-arginine is from about 15°C to about 83°C. In some embodiments, the temperature of the salt-forming mixture at the time of addition of L-arginine is from about 18°C to about 80°C. In some embodiments, the temperature of the salt-forming mixture at the time of addition of L-arginine is from about 20°C to about 40°C. In some embodiments, the temperature of the salt-forming mixture at the time of addition of L-arginine is from about 18°C to about 30°C.

In some embodiments, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine in the form of an aqueous solution.

In some embodiments of the present invention, the addition of L-arginine is accomplished by adding L-arginine to the salt-forming mixture in the form of a solution, wherein the L-arginine solution is an aqueous solution of a concentration of from 2.1 M to 2.3 M at a temperature of from about 50°C up to approximately 75°C.

- 19 045975

In some embodiments of the present invention, the production of the first mixture in step b) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments of the present invention, the production of the first mixture in step b) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments of the present invention, the production of the first mixture in step b) is carried out at a mixing speed of about 150 rpm.

Step c) - Heating the first mixture to the temperature of the first heating to obtain the second mixture

In some embodiments of the present invention, the first heating temperature is from about 20°C to about 85°C. In some embodiments, the first heating temperature is from about 20°C to about 83°C. In some embodiments, the first heating temperature is from about 20°C to about 55°C. In some embodiments, the first heating temperature is from about 25°C to about 45°C. In some embodiments, the first heating temperature is from about 50°C to about 83°C. In some embodiments, the first heating temperature is from about 60°C to about 80°C. In some embodiments, the first heating temperature is from about 70°C to about 80°C. In some embodiments, the first heating temperature is from about 79°C to about 85°C.

In some embodiments of the present invention, additional H ₂ O is optionally added to the second mixture.

In some embodiments of the present invention, when additional H2O is optionally added to the second mixture, the weight of water added is from about 0.4 to about 2.3 times the weight of L-arginine originally added to form the first mixture.

In some embodiments of the present invention, when additional H2O is optionally added to the second mixture, the weight ratio of water added to L-Arginine originally added to produce the first mixture is from about 1.0:1.6 to about 1.0:2.1 . In some embodiments, when H2O is optionally added to the second mixture, the weight ratio of water added to L-arginine originally added to form the first mixture is from about 1.0:0.8 to about 1.0:1.9. In some embodiments, when additional H ₂ O is optionally added to the second mixture, the weight ratio of the amount of water added to the amount of L-arginine initially added to produce the first mixture is from about 1.0:1.0 to about 1.0 :1.7. When additional H2O is optionally added to the second mixture, the weight ratio of the water added to the L-arginine originally added to form the first mixture is from about 1.0:1.1 to about 1.0:1.6. In some embodiments, when additional H2O is optionally added to the second mixture, the weight ratio of water added to L-arginine originally added to produce the first mixture is from about 1.0:1.2 to about 1.0:1.5 .

In some embodiments of the present invention, the second mixture is essentially a homogeneous solution.

In some embodiments, step c) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, step c) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step c) is carried out at a mixing speed of approximately 150 rpm.

Step d) - Adding a first additional amount of water-miscible antisolvent to the second mixture while maintaining the first heating temperature to obtain a suspension

In some embodiments of the present invention, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:5.46 to about 1.00:8.20.

In some embodiments of the present invention, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:4.70 to about 1.00:7.50. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid and the first additional amount of 2-propanol is from about 1.00:5.20 to about 1.00:7.00. In some embodiments, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid and the first additional amount of 2-propanol is from about 1.00:5.70 to about 1.00:6.50. In some embodiments, the weight ratio of L-arginine salt is L-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from

- 20 045975 approximately 1.00:5.90 to approximately 1.00:6.30. In some embodiments, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid and the first additional amount of 2-propanol is from about 1.00:5.95 to about 1.00:6.25. In some embodiments, the weight ratio of L-arginine salt is L-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:6.00 to about 1.00:6.20.

In some embodiments of the present invention, the weight ratio of L-arginine salt is L)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:5.46 to about 1.00:8.20, from about 1.00:5.81 to about 1.00:7.86, from about 1.00:6.15 to about 1.00:7.51, from about 1.00:6.28 to about 1.00:7.38, from about 1.00:6.42 to about 1.00 :7.24, from about 1.00:6.56 to about 1.00:7.10, from about 1.00:6.69 to about 1.00:6.97 or about 1.00:6, 83.

In some embodiments of the present invention, a first additional amount of water-miscible antisolvent is added at a first time point and a second time point.

In some embodiments of the present invention, from about 5% to about 15% of the first additional amount of water-miscible antisolvent is added at a first time point.

In some embodiments, from about 7% to about 13% of the first additional amount of water-miscible antisolvent is added at the first time point.

In some embodiments, from about 8% to about 12% of the first additional amount of water-miscible antisolvent is added at a first time point. In some embodiments, about 9% to about 11% of the first additional amount of water-miscible antisolvent is added at a first time point.

In some embodiments, a first additional amount of water-miscible antisolvent is added at a first time to produce a cloudy mixture.

In some embodiments of the present invention, a seed crystal of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] is optionally added prior to the second time point. indol-3-yl)acetic acid.

In some embodiments of the present invention, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed in about 1.00 hour or more. In some embodiments, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed within about 1.00 hours to about 6.00 hours. In some embodiments, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed within about 1.00 to about 5.00 hours. In some embodiments, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed within about 1.00 to about 4.00 hours. In some embodiments, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed within about 1.00 to about 3.00 hours. In some embodiments, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed within about 1.00 to about 2.00 hours.

In some embodiments of the present invention, the first heating temperature is maintained while the first additional amount of 2-propanol is added to the second mixture.

In some embodiments, step d) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, step d) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step d) is carried out at a mixing speed of about 150 rpm.

Stage e) - Cooling the suspension to the first cooling temperature and subsequent heating to the second heating temperature

In some embodiments of the present invention, cooling the slurry to the first cooling temperature in step e) is carried out at a rate of from about 8.80°C/hour to about 14.40°C/hour. In some embodiments, cooling the slurry to the first cooling temperature in step e) is carried out at a rate of from about 9.35°C/hour to about 13.80°C/hour. In some embodiments, cooling the slurry to the first cooling temperature in step e) is carried out at a rate of from about 9.90°C/hour to about 13.20°C/hour. In some variants

- 21 045975 cooling the suspension to the first cooling temperature in step e) is carried out at a rate of from about 10.45°C/hour to about 12.60°C per hour. In some embodiments, cooling the slurry to the first cooling temperature in step e) is carried out at a rate of from about 10°C/hour to about 12°C/hour. In some embodiments, cooling the slurry to the first cooling temperature in step e) is carried out at a rate of from about 9°C/hour to about 11°C/hour. In some embodiments, cooling the slurry to the first cooling temperature in step e) is carried out at a rate of from about 9.5°C/hour to about 10.5°C/hour.

In some embodiments of the present invention, the first cooling temperature in step e) is from about 15°C to about 40°C. In some embodiments, the first cooling temperature in step e) is from about 20°C to about 30°C. In some embodiments, the first cooling temperature in step e) is from about 22°C to about 24°C. In some embodiments, the first cooling temperature in step e) is from about 18°C to about 22°C.

In some embodiments of the present invention, the first cooling temperature is maintained for at least 1 hour before heating to the second heating temperature.

In some embodiments of the present invention, the second heating temperature in step e) is from about 65°C to about 83°C. In some embodiments, the second heating temperature in step e) is from about 70°C to about 80°C. In some embodiments, the second heating temperature in step e) is from about 70°C to about 75°C. In some embodiments, the second heating temperature in step e) is from about 69°C to about 73°C.

In some embodiments of the present invention, after heating to the second heating temperature, the second heating temperature is maintained for at least 30 minutes.

In some embodiments, step e) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, step e) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step e) is carried out at a mixing speed of approximately 150 rpm.

Stage f) - Cyclic repetition of stage e) optionally one or more times

It should be understood that when cyclically repeating step e) more than once, the first cooling temperature of each cycle may be the same or different, and the said second heating temperature of each cycle may be the same or different.

Two cycles

In some embodiments of the present invention, cycling step f) involves cycling step e) twice.

In some embodiments of the present invention, cycling step e) twice comprises cooling the slurry to a first cycle cooling temperature, heating the slurry to a first cycle heating temperature, cooling the slurry to a second cycle cooling temperature, and heating the slurry to a second cycle heating temperature.

In some embodiments of the present invention, the first cycle cooling temperature is from about 16°C to about 26°C, the first cycle heating temperature is from about 55°C to about 65°C, and the second cycle cooling temperature is from about 26°C to about 36°C, and the heating temperature of the second cycle is from about 45°C to about 55°C. In some embodiments, the first cycle cooling temperature is from about 19°C to about 23°C, the first cycle heating temperature is from about 58°C to about 62°C, and the second cycle cooling temperature is from about 29°C to about 33°C. C, and the heating temperature of the second cycle is from about 48°C to about 52°C.

In some embodiments of the present invention, cooling to the cooling temperature of each cycle is conducted at substantially different cooling rates. In some embodiments, cooling to the cooling temperature of each cycle is conducted at a different cooling rate.

In some embodiments of the present invention, cycling step f) involves cycling step e) twice. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 10.0°C/hour to about 15.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 11.0°C/hour to about 14.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 12.0°C/hour to about 13.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of about 12.5°C/hour.

In some embodiments, cycling step f) includes looping step e) twice. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 7.5°C/hour to about 12.5°C/hour. Somehow

- 22 045975 In some embodiments, cooling to the second cycle cooling temperature is carried out at a rate of from about 8.5°C/hour to about 11.5°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 9.5°C/hour to about 10.5°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of about 10.0° C./hour.

Three cycles

In some embodiments of the present invention, cycling step f) includes cycling step e) three times.

In some embodiments of the present invention, cycling step e) three times comprises cooling the suspension to the first cycle cooling temperature, heating the suspension to the first cycle heating temperature, cooling the suspension to the second cycle cooling temperature, heating the suspension to the second cycle heating temperature, cooling the suspension to the cooling temperature the third cycle and heating the suspension to the heating temperature of the third cycle.

In some embodiments, the first cycle cooling temperature is from about 16°C to about 26°C, the first cycle heating temperature is from about 66°C to about 76°C, and the second cycle cooling temperature is from about 16°C to about 26°C. C, the second cycle heating temperature is from about 55°C to about 65°C, the third cycle cooling temperature is from about 26°C to about 36°C, and the third cycle heating temperature is from about 45°C to about 55°C. In some embodiments, the first cycle cooling temperature is from about 19°C to about 23°C, the first cycle heating temperature is from about 69°C to about 73°C, and the second cycle cooling temperature is from about 19°C to about 23°C. C, the second cycle heating temperature is from about 58°C to about 62°C, the third cycle cooling temperature is from about 29°C to about 33°C, and the third cycle heating temperature is from about 48°C to about 52°C.

In some embodiments of the present invention, cooling to the cooling temperature of each cycle is carried out at a substantially different cooling rate. In some embodiments, cooling to the cooling temperature of each cycle is performed at a different cooling rate.

In some embodiments, cooling to the first cycle cooling temperature and cooling to the second cycle cooling temperature are conducted at substantially the same cooling rate.

In some embodiments of the present invention, cycling step f) includes cycling step e) three times. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 10.0°C/hour to about 15.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 11.0°C/hour to about 14.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 12.0°C/hour to about 13.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of about 12.5°C/hour.

In some embodiments of the present invention, cycling step f) includes cycling step e) three times. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 10.0°C/hour to about 15.0°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 11.0°C/hour to about 14.0°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 12.0°C/hour to about 13.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of from about 12.5°C/hour.

In some embodiments of the present invention, cycling step f) includes cycling step e) three times. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 7.5°C/hour to about 12.5°C/hour. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 8.5°C/hour to about 11.5°C/hour. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 9.5°C/hour to about 10.5°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of about 10.0° C./hour.

Four cycles

In some embodiments of the present invention, cycling step f) involves cycling step e) four times.

In some embodiments, cycling step e) four times includes cooling the slurry to the first cycle cooling temperature, heating the slurry to the first cycle heating temperature, cooling the suspension to the second cycle cooling temperature, heating the suspension to the second cycle heating temperature, cooling the suspension to the third cycle cooling temperature

- 23 045975 cycle, heating the suspension to the heating temperature of the third cycle, cooling the suspension to the cooling temperature of the fourth cycle and heating the suspension to the heating temperature of the fourth cycle.

In some embodiments of the present invention, the first cycle cooling temperature is from about 16°C to about 26°C, the first cycle heating temperature is from about 66°C to about 76°C, and the second cycle cooling temperature is from about 16°C to about 26°C, second cycle heating temperature is from about 66°C to about 76°C, third cycle cooling temperature is from about 16°C to about 26°C, third cycle heating temperature is from about 55°C to about 65°C C, the fourth cycle cooling temperature is from about 26°C to about 36°C, the fourth cycle heating temperature is from about 45°C to about 55°C. In some embodiments, the first cycle cooling temperature is from about 19°C to about 23°C, the first cycle heating temperature is from about 69°C to about 73°C, and the second cycle cooling temperature is from about 19°C to about 23°C. C, the second cycle heating temperature is from about 69°C to about 73°C, the third cycle cooling temperature is from about 19°C to about 23°C, the third cycle heating temperature is from about 58°C to about 62°C, the fourth cycle cooling temperature is from about 29°C to about 33°C, the fourth cycle heating temperature is from about 48°C to about 52°C.

In some embodiments of the present invention, cooling to the cooling temperature of each cycle is carried out at a substantially different cooling rate. In some embodiments, cooling to the cooling temperature of each cycle is performed at a different cooling rate.

In some embodiments of the present invention, cooling to the first cycle cooling temperature, to the second cycle cooling temperature, and to the third cycle cooling temperature is carried out at substantially the same cooling rate.

In some embodiments, the cycling of step f) involves cycling step e) four times. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 10.0°C/hour to about 15.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 11.0°C/hour to about 14.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 12.0°C/hour to about 13.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of about 12.5°C/hour.

In some embodiments of the present invention, cycling step f) involves cycling step e) four times. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 10.0°C/hour to about 15.0°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 11.0°C/hour to about 14.0°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 12.0°C/hour to about 13.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of about 12.5°C/hour.

In some embodiments of the present invention, cycling step f) involves cycling step e) four times. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 10.0°C/hour to about 15.0°C/hour. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 11.0°C/hour to about 14.0°C/hour. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 12.0°C/hour to about 13.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of about 12.5°C/hour.

In some embodiments of the present invention, cycling step f) involves cycling step e) four times. In some embodiments, cooling to the fourth cycle cooling temperature is conducted at a rate of from about 7.5°C/hour to about 12.5°C/hour. In some embodiments, cooling to the fourth cycle cooling temperature is conducted at a rate of from about 8.5°C/hour to about 11.5°C/hour. In some embodiments, cooling to the fourth cycle cooling temperature is conducted at a rate of from about 9.5°C/hour to about 10.5°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of about 10.0° C./hour.

In some embodiments, step f) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, step f) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step f) is carried out at a mixing speed of approximately 150 rpm.

Step g) - Cooling the suspension to the final cooling temperature to obtain free

- 24 045975 lamellar crystalline form of L-arginine salt U)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indol-3-yl) acetic acid

In some embodiments of the present invention, the cooling of the suspension in step g) is carried out at a rate of from about 7.5°C/hour to about 12.5°C/hour. In some embodiments, the cooling of the suspension in step g) is carried out at a rate of from about 8.5°C/hour to about 11.5°C/hour. In some embodiments, the cooling of the suspension in step g) is carried out at a rate of from about 9.5°C/hour to about 10.5°C/hour. In some embodiments, the cooling of the suspension in step g) is carried out at a rate of about 10.0°C/hour.

In some embodiments of the present invention, the cooling of the suspension in step g) is carried out at a rate of from about 8.8°C/hour to about 14.4°C/hour. In some embodiments, the cooling of the slurry in step g) is carried out at a rate of from about 9.4°C/hour to about 13.8°C/hour. In some embodiments, the cooling of the slurry in step g) is carried out at a rate of from about 9.9°C/hour to about 13.2°C/hour. In some embodiments, the cooling of the slurry in step g) is carried out at a rate of from about 10.5°C/hour to about 12.6°C/hour. In some embodiments, the cooling of the suspension in step g) is carried out at a rate of from about 10°C/hour to about 12°C/hour.

In some embodiments of the present invention, after cooling in step g), the temperature of the suspension is from about 15°C to about 40°C. In some embodiments, after cooling in step g), the temperature of the slurry is from about 20°C to about 30°C. In some embodiments, after cooling in step g), the temperature of the slurry is from about 22°C to about 24°C. In some embodiments, after cooling in step g), the temperature of said suspension is from about 18°C to about 22°C.

In some embodiments, step g) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, step g) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step g) is carried out at a mixing speed of approximately 150 rpm.

In some embodiments, the method further includes the step of isolating the free-lamellar crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid.

In some embodiments of the present invention, the isolation step comprises filtering the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b ]indol-3-yl)acetic acid from the suspension. In some embodiments, the isolation step comprises filtering the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid from the suspension and drying the free-plate crystalline form of L-arginine salt under reduced pressure.

In some embodiments of the present invention, after filtration, the free-plate crystalline form of the L-arginine salt is A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indole-3 -yl)acetic acid has one or more properties that are described herein. For example, in some embodiments, after filtration, the onset temperature of the endothermic transition of the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl)acetic acid is in the range from 205.0°C to 208.5°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtration, the temperature at which the free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy))-1,2,3,4-tetrahydrocyclopentate [b ]indol-3-yl)acetic acid is in the range from 205.5°C to 208.5°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtration, the onset temperature of the endothermic transition of the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid is in the range from 206.5°C to 208.5°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtration, the temperature at which the free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy))-1,2,3,4-tetrahydrocyclopentate [b ]indol-3-yl)acetic acid is in the range from 207.0°C to 208.1°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtration on the differential scanning calorimetry curve of the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid obtained

- 25 045975 when scanning at a rate of 10°C/min, there is an endotherm as substantially represented in any of Figures 6 to 10. In some embodiments, after filtering on the differential scanning calorimetry curve of the free-plate crystalline form of L-arginine salt ( R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid obtained by scanning at a speed of 10° C/min, there is an endotherm, as substantially represented in any of Figures 6 to 10 and 22 to 25. In some embodiments, after filtration, a plot of dynamic moisture sorption (DMS) of the free-plate crystalline form of L-arginine salt (K .)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclic lopenta[b]indol-3-yl)acetic acid with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-plate crystalline form of about 0.3% by weight or less at 90% RH. In some embodiments, after filtration, the dynamic moisture sorption (DMS) plot of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid with an adsorption phase from 30% RH to 90% RH shows an increase in the weight of the free-lamellar crystalline form of about 0.2% by weight or less at 90% RH. The DSW characteristics provided in the present invention may also vary by plus/minus 0.15% change in mass (ie, ±0.15% change in mass).

In some embodiments of the present invention, after filtration, a plot of the total impurities in the free-plate crystalline form of L-arginine salt (R)-2 -(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid obtained by achiral HPLC shows that the total impurity content is 0.5 area% or less. In some embodiments, after filtration, a plot of the total impurities in the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid obtained by achiral HPLC shows that the total impurity content is 0.3 area% or less. In some embodiments of the present invention, after filtration, a plot of the total impurities in the free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid obtained by achiral HPLC shows a total impurity content of 0.2 area% or less.

In some embodiments of the present invention, the method further includes the step of preparing a free-plate crystalline form of L-arginine salt (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid to obtain a pharmaceutical composition.

In some embodiments of the present invention, the method further includes the step of mixing the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid with a pharmaceutical excipient to obtain a pharmaceutical composition.

In some embodiments of the present invention, the method further includes the step of mixing the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid with a pharmaceutical excipient to prepare a pharmaceutical composition suitable for oral, rectal, nasal, topical, buccal, sublingual or vaginal administration, or in a form suitable for administration by inhalation, insufflation or transdermal patches . In some embodiments, the pharmaceutical composition is suitable for oral administration.

In addition, the present invention describes methods for preparing the free-plate crystalline form of L-arginine salt (K)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[B] indol-3-yl)acetic acid, based on various morphologies, such as, without limitation, spherulites/radial clusters, fine particles, agglomerates, flakes, or mixtures thereof.

Accordingly, one aspect of the present invention relates to a method for preparing a free-plate crystalline form of L-arginine salt (K)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b ]indol-3-yl)acetic acid, where said method includes the following steps:

a) heating the first mixture containing L-arginine salt (K)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3- yl)acetic acid, water-miscible antisolvent and water, to the temperature of the first heating to obtain the second mixture;

b) cooling the second mixture to the first cooling temperature, then adding a first additional amount of water-miscible antisolvent to the second mixture while maintaining the first cooling temperature, and then heating to the second heating temperature to obtain a suspension;

c) cyclically repeating step b) optionally once or twice; And

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d) cooling the suspension to a second cooling temperature to obtain a free-plate crystalline form of L-arginine salt(s)-2-(7-(4-cyclopentyl-3-(lfluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid.

In some embodiments of the present invention, the water miscible antisolvent includes a solvent selected from the group consisting of acetonitrile, acetone, tetrahydrofuran and C ₂ -C ₄ alkanol. In some embodiments, the water-miscible antisolvent includes a solvent selected from the group consisting of acetonitrile, acetone, tetrahydrofuran, ethanol, 1-propanol, 2-propanol, and 1-butanol. In some embodiments, the water-miscible antisolvent includes 2-propanol.

In some embodiments of the present invention, L-arginine salt of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indol-3-yl)acetic acid at stage a) is a spherulite, radial cluster, small particles, agglomerate, flakes or a mixture thereof.

In some embodiments of the present invention, L-arginine salt of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indol-3-yl)acetic acid in step a) is a spherulitic crystalline form (a crystalline form with a spherulitic crystal habit) or a mixture containing a crystalline spherulitic form.

In some embodiments of the present invention, a method for preparing a free-plate crystalline form of L-arginine salt (L)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indole -3-yl)acetic acid is carried out in an inert atmosphere. In some embodiments, the method is conducted under an inert atmosphere containing nitrogen or argon. In some embodiments, the method is conducted under an inert atmosphere containing nitrogen.

Stage a) - Heating the first mixture

In some embodiments of the present invention, the first mixture contains L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl )acetic acid, 2-propanol and water in a mass ratio of from about 1.00:3.35:0.75 to about 1.00:5.03:1.13. In some embodiments, the first mixture contains L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl)acetic acid, 2-propanol and water in a weight ratio of from about 1.00:3.56:0.80 to about 1.00:4.82:1.08. In some embodiments, the first mixture contains L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl)acetic acid, 2-propanol and water in a weight ratio of from about 1.00:3.77:0.85 to about 1.00:4.61:1.03. In some embodiments, the first mixture contains L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl)acetic acid, 2-propanol and water in a weight ratio of from about 1.00:3.85:0.86 to about 1.00:4.52:1.02. In some embodiments, the first mixture contains L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl)acetic acid, 2-propanol and water in a mass ratio of about 1.00:3.94:0.88 to 1.00:4.44:1.00. In some embodiments, the first mixture contains L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid, 2-propanol and water in a weight ratio of from about 1.00:4.02:0.90 to about 1.00:4.36:0.98. In some embodiments, the first mixture contains L-arginine salt (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid, 2-propanol and water in a weight ratio of from about 1.00:4.11:0.92 to about 1.00:4.27:0.96. In some embodiments, the first mixture contains L-arginine salt (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid, 2-propanol and water in a mass ratio of approximately 1.00:4.19:0.94.

In some embodiments of the present invention, the temperature of the first mixture is from about 10°C to about 30°C before heating to the first heating temperature. In some embodiments, the temperature of the first mixture is from about 15°C to about 25°C before heating to the first heating temperature.

In some embodiments of the present invention, the first heating temperature is from about 60°C to about 83°C. In some embodiments, the first heating temperature is from about 70°C to about 83°C. In some embodiments, the first heating temperature is from about 75°C to about 80°C.

In some embodiments, after heating to the first heating temperature, the second mixture is a substantially homogeneous solution.

In some embodiments, step a) is carried out at a mixing speed of from about 50 rpm to about 250 rpm. In some embodiments, step a) is carried out at a mixing speed of from about 75 rpm to about 150 rpm.

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Stage b) - cooling the second mixture to the temperature of the first cooling

In some embodiments of the present invention, the first cooling temperature is from about 50°C to about 70°C. In some embodiments, the first cooling temperature is from about 60°C to about 70°C. In some embodiments, the first cooling temperature is from about 63°C to about 67°C.

In some embodiments of the present invention, the first cooling temperature is maintained for at least 30 minutes.

In some embodiments of the present invention, the addition of the first additional amount of 2-propanol to the second mixture is carried out at a rate of from about 107.20 g/min to about 160.80 g/min. In some embodiments, the addition of the first additional amount of 2-propanol to the second mixture is carried out at a rate of from about 113.90 g/min to about 154.10 g/min. In some embodiments, the addition of the first additional amount of 2-propanol to the second mixture is carried out at a rate of from about 120.60 g/min to about 147.40 g/min. In some embodiments, the addition of the first additional amount of 2propanol to the second mixture is carried out at a rate of from about 127.30 g/min to about 140.70 g/min. In some embodiments, the addition of the first additional amount of 2propanol to the second mixture is carried out at a rate of about 134 g/min.

In some embodiments of the present invention, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:0.98 to about 1.00:1.47. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indol-3-yl)acetic acid and the first additional amount of 2-propanol is from about 1.00:1.04 to about 1.00:1.41. In some embodiments of the present invention, the weight ratio of L-arginine salt is ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid and the first additional amount of 2propanol is from about 1.00:1.10 to about 1.00:1.35. In some embodiments of the present invention, the weight ratio of L-arginine salt is L-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:1.13 to about 1.00:1.32. In some embodiments of the present invention, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3 yl )acetic acid and the first additional amount of 2-propanol is from about 1.00:1.15 to about 1.00:1.30. In some embodiments of the present invention, the weight ratio of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[L]indol-3-yl)acetic acid and the first additional amount of 2propanol is from about 1.00:1.18 to about 1.00:1.27. In some embodiments of the present invention, the weight ratio of L-arginine salt is L-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:1.20 to about 1.00:1.25. In some embodiments of the present invention, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3 - yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:1.23.

In some embodiments of the present invention, after adding the first additional amount of 2-propanol, the first cooling temperature is maintained for at least 1 hour before heating to the second heating temperature.

In some embodiments of the present invention, the second heating temperature is from about 65°C to about 83°C. In some embodiments, the second heating temperature is from about 70°C to about 80°C. In some embodiments, the second heating temperature is from about 70°C to about 75°C.

In some embodiments of the present invention, the second heating temperature is maintained to maintain the suspension.

In some embodiments, after heating to the second heating temperature, the second heating temperature is maintained for at least 30 minutes to maintain the suspension.

In some embodiments, step b) is carried out at a mixing speed of from about 50 rpm to about 150 rpm. In some embodiments, step b) is carried out at a mixing speed of from about 70 rpm to about 110 rpm.

Stage c) - Cyclic repetition of stage b)

In some embodiments of the present invention, step b) is cycled once.

In some embodiments of the present invention, cyclic repetition of step b)

- 28 045975 is carried out once, where the cyclic repetition is carried out essentially under the conditions used in step b).

In some embodiments of the present invention, the cycling of step b) is performed once, where the cycling conditions are different from the conditions used in step b); It should be understood that the cycling conditions selected are within the conditions of the embodiments described herein for step b).

In some embodiments of the present invention, step b) is repeated twice.

In some embodiments of the present invention, the cycling of step b) is performed twice, where the cycling conditions of the first cycle and the second cycle are essentially the conditions used in step b).

In some embodiments of the present invention, step b) is cycled twice, where at least one of the conditions of the first cycle, the second cycle or step b) is different; it should be understood that regardless of whether the first cycle and/or the second cycle are different from each other or one or both cycles from step b), each of the selected conditions for the first and second cycles is within the options described herein for stage b).

Step d) - Cooling the suspension to the second cooling temperature to obtain the free-plate crystalline form of L-arginine salt (D)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid

In some embodiments of the present invention, the second cooling temperature is from about 50°C to about 70°C. In some embodiments, the second cooling temperature is from about 60°C to about 70°C. In some embodiments, the second cooling temperature is from about 63°C to about 67°C.

In some embodiments of the present invention, the second cooling temperature is maintained for at least 30 minutes.

In some embodiments of the present invention, step d) is carried out at a mixing speed of from about 25 revolutions per minute to about 105 revolutions per minute. In some embodiments, step d) is carried out at a mixing speed of from about 45 revolutions per minute to about 85 revolutions per minute.

In some embodiments of the present invention, after cooling to the second cooling temperature, the method further includes the step of cooling the suspension to the third cooling temperature.

In some embodiments of the present invention, after cooling to a second cooling temperature, the method further includes the step of cooling the slurry to a third cooling temperature of from about 10°C to about 30°C. In some embodiments, after cooling to a second cooling temperature, the method further includes the step of cooling the slurry to a third cooling temperature of from about 15°C to about 25°C.

In some embodiments of the present invention, the cooling rate from the second cooling temperature to the third cooling temperature is from about 5°C per hour to about 15°C per hour. In some embodiments, the cooling rate from the second cooling temperature to the third cooling temperature is from about 8°C per hour to about 12°C per hour. In some embodiments, the cooling rate from the second cooling temperature to the third cooling temperature is approximately 10°C per hour.

In some embodiments of the present invention, the method further includes the step of isolating the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid.

In some embodiments of the present invention, the isolation step includes filtration from the suspension of the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid. In some embodiments, the isolation step comprises filtering from the suspension the free-plate crystalline form of L-arginine salt (1-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl)acetic acid by filtration and drying the free-plate crystalline form of L-arginine salt under reduced pressure.

In some embodiments of the present invention, after filtration, the onset temperature of the phase transition of the crystalline form of L-arginine salt (D)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L] ]indol-3-yl)acetic acid is in the range from 205.0°C to 208.5°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtration, the onset temperature of the phase transition of the crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol- 3-yl)acetic acid is in the range from 205.5°C to 208.5°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtration, the onset temperature of the phase transition of the crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid is in the range from 206.5°C to 208.5°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtration, the onset temperature of the phase transition of the crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol- 3-yl)acetic acid is in the range from 207.0°C to 208.1°C, as determined by differential scanning calorimetry when scanning at a speed of 10°C/min. In some embodiments, after filtration on the differential scanning calorimetry curve of L-arginine salt, ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid obtained by scanning at a rate of 10° C./min. has an endotherm as substantially represented in any of Figures 6 to 10. In some embodiments, after filtering on the L-arginine differential scanning calorimetry curve salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid obtained by scanning at speed 10°C/min, there is an endotherm, as substantially represented in any of Figures 6 to 10 and 22 to 25. In some embodiments, after filtration, a plot of dynamic moisture sorption (DMS) of the free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3 -(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of about 0.3% by weight or less at 90% RH. In some embodiments, after filtration, the dynamic moisture sorption (DMS) plot of the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of about 0.2% by weight or less at 90% RH. The DSW characteristics provided in the present invention may also vary by plus/minus 0.15% change in mass (ie, ±0.15% change in mass).

In some embodiments of the present invention, after filtration, a plot of the total impurities in the free-plate crystalline form of L-arginine salt (R)-2-(7(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4 -tetrahydrocyclopenta[b]indol-3-yl)acetic acid obtained by achiral HPLC shows a total impurity content of 0.5 area%. In some embodiments, after filtration, a plot of the total impurities in the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid obtained by achiral HPLC shows that the total impurity content is 0.3% by area. In some embodiments, after filtration, a plot of the total impurities in the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid obtained by achiral HPLC shows a total impurity content of 0.2% by area.

In some embodiments of the present invention, the method further includes the step of preparing a free-plate crystalline form of L-arginine salt (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid to obtain a pharmaceutical composition.

In some embodiments of the present invention, the method further includes the step of mixing the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid with a pharmaceutical excipient to obtain a pharmaceutical composition.

Another aspect of the present invention relates to a method for preparing the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, where said method includes the following steps:

a) obtaining the first mixture containing L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid, water-miscible antisolvent and H2O;

b) heating the first mixture to the first heating temperature to obtain a second mixture;

c) adding a first additional amount of water-miscible antisolvent to the second mixture while maintaining the first heating temperature to obtain a suspension;

d) cooling the suspension to the first cooling temperature and then heating to the second heating temperature;

- 30 045975

e) cycling step d) optionally one or more times, where the first cooling temperature of each cycle may be the same or different, and the second heating temperature of each cycle may be the same or different; And

e) cooling the suspension to the final cooling temperature to obtain the free-plate crystalline form of L-arginine salt 4)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid.

Stage a) - Preparation of the first mixture

In some embodiments of the present invention, the preparation of the first mixture in step a) is carried out under an inert atmosphere. In some embodiments, the preparation of the first mixture in step a) is carried out under an inert atmosphere containing argon or nitrogen. In some embodiments of the present invention, the preparation of the first mixture in step a) is carried out under an inert atmosphere containing nitrogen.

It should be borne in mind that the preparation of the first mixture containing L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3 -yl)acetic acid, water-miscible antisolvent and H2O can be performed in any manner conventional to one skilled in the art, such as those described above.

In some embodiments of the present invention, preparation of the first mixture in step a) includes the step of adding L-arginine and H2O, together or separately in any order, to the salt-forming mixture containing 4)-2-(7-(4-cyclopentyl-3- (trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol-3-yl)acetic acid and a water-miscible antisolvent to obtain the first mixture.

In some embodiments of the present invention, the addition of L-arginine and H2O is carried out under an inert atmosphere. In some embodiments, the addition of L-arginine and H2O is carried out under an inert atmosphere containing argon or nitrogen. In some embodiments, the addition of L-arginine and H2O is carried out under an inert atmosphere containing nitrogen.

In some embodiments of the present invention, the water-miscible antisolvent includes a solvent selected from the group consisting of acetonitrile, acetone, tetrahydrofuran, and N-alkanol. In some embodiments, the water-miscible antisolvent includes a solvent selected from the group consisting of acetonitrile, acetone, tetrahydrofuran, ethanol, 1-propanol, 2-propanol, and 1-butanol. In some embodiments, the water-miscible antisolvent includes 2-propanol.

In some embodiments of the present invention, before adding L-arginine and H2O, the salt-forming mixture contains 4)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indole-3 -yl)acetic acid and 2-propanol in a weight ratio of from about 1.0:3.0 to about 1.0:11.0. In some embodiments, before adding L-Arginine and H2O, the salt-forming mixture contains 4)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid and 2-propanol in a weight ratio of from about 1.0:4.0 to about 1.0:10.0. In some embodiments, before adding L-Arginine and H2O, the salt-forming mixture contains 4)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid and 2-propanol in a weight ratio of from about 1.0:5.0 to about 1.0:9.0. In some embodiments, before adding L-Arginine and H2O, the salt-forming mixture contains 4)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid and 2-propanol in a weight ratio of from about 1.0:6.0 to about 1.0:8.0.

In some embodiments of the present invention, the molar ratio of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L -arginine is from about 1.0:0.8 to about 1.0:1.2.

In some embodiments of the present invention, the molar ratio of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L -arginine is from about 1.0:0.9 to about 1.0:1.2. In some embodiments, the molar ratio of 4)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol-3-yl)acetic acid to L-arginine is from about 1.0:1.0 to about 1.0:1.2. In some embodiments, the molar ratio of (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L-arginine ranges from about 1.0:0.93 to about 1.0:1.01. In some embodiments, the molar ratio of 4)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol-3-yl)acetic acid to L-arginine is from about 1.0:0.93 to about 1.0:0.97. In some embodiments of the present invention, the molar ratio of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3yl)acetic acid and L-arginine is approximately 1.0:1.0. In some embodiments, the molar ratio of 4)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L -arginine is approximately 1.0:0.95.

- 31 045975

In some embodiments of the present invention, the weight ratio of L-arginine to H2O is from about 1.0:0.4 to about 1.0:2.3. In some embodiments, the weight ratio of L-arginine to H2O is from about 1.0:1.6 to about 1.0:2.1. In some embodiments, the weight ratio of L-arginine to H2O is from about 1.0:0.8 to about 1.0:1.9. In some embodiments, the weight ratio of L-arginine to H2O is from about 1.0:1.0 to about 1.0:1.7. In some embodiments, the weight ratio of L-arginine to H2O is from about 1.0:1.1 to about 1.0:1.6. In some embodiments, the weight ratio of L-arginine to H2O is from about 1.0:1.2 to about 1.0:1.5.

In some embodiments of the present invention, the temperature of the salt-forming mixture before adding L-arginine is from about 15°C to about 83°C. In some embodiments, the temperature of the salt-forming mixture before adding L-arginine is from about 18°C to about 80°C. In some embodiments, the temperature of the salt-forming mixture before adding L-arginine is from about 20°C to about 40°C. In some embodiments, the temperature of the salt-forming mixture before adding L-arginine is from about 18°C to about 30°C.

In some embodiments of the present invention, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine in the form of an aqueous suspension.

In some embodiments, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine as a solid.

In some embodiments of the present invention, the addition of L-arginine to the salt-forming mixture is accomplished by adding substantially all of the L-arginine as a solid to the salt-forming mixture. The phrase essentially all at once or all at once refers to the addition of the entire amount of L-arginine to the salt-forming mixture at one time, with the only limitation being the equipment used.

In some embodiments of the present invention, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine as a solid to the salt-forming mixture over a period of about 30 minutes. In some embodiments, the addition of L-Arginine to the salt-forming mixture is accomplished by adding L-arginine as a solid to the salt-forming mixture over a period of about 1 hour. In some embodiments, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine as a solid to the salt-forming mixture over a period of about 2 hours.

In some embodiments of the present invention, the salt-forming mixture contains (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3yl)acetic acid , 2-propanol and water in a mass ratio of from about 1.0:3.0:0.05 to about 1.0:11.0:1.0. In some embodiments, the salt-forming mixture contains (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, 2- propanol and water in a mass ratio of from about 1.0:4.0:0.1 to about 1.0:10.0:0.9. In some embodiments of the present invention, the salt-forming mixture contains (R)-2-(7(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid , 2-propanol and water in a mass ratio of from about 1.0:5.0:0.15 to about 1.0:9.0:0.8. In some embodiments of the present invention, the salt-forming mixture contains (R)-2-(7(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid , 2-propanol and water in a mass ratio of from about 1.0:6.0:0.25 to about 1.0:8.0:0.7.

In some embodiments of the present invention, the temperature of the salt-forming mixture upon addition of L-arginine is from about 15°C to about 83°C. In some embodiments, the temperature of the salt-forming mixture upon addition of L-arginine is from about 18°C to about 80°C. In some embodiments, the temperature of the salt-forming mixture upon addition of L-arginine is from about 20°C to about 40°C. In some embodiments, the temperature of the salt-forming mixture upon addition of L-arginine is from about 18°C to about 30°C.

In some embodiments of the present invention, the temperature of the salt-forming mixture upon addition of L-arginine is from about 15°C to about 83°C. In some embodiments, the temperature of the salt-forming mixture upon addition of L-arginine is from about 18°C to about 80°C. In some embodiments, the temperature of the salt-forming mixture upon addition of L-arginine is from about 20°C to about 40°C. In some embodiments, the temperature of the salt-forming mixture upon addition of L-arginine is from about 18°C to about 30°C.

In some embodiments of the present invention, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine in the form of an aqueous solution.

In some embodiments, the addition of L-arginine to the salt-forming mixture is accomplished by adding L-arginine in the form of an aqueous solution, wherein the aqueous solution of L-arginine is

- 32 045975 is an aqueous solution of concentration from 2.1 M to 2.3 M with a temperature from about 50°C to about 75°C.

In some embodiments of the present invention, the preparation of the first mixture in step a) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, the preparation of the first mixture in step a) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments of the present invention, the production of the first mixture in step a) is carried out at a mixing speed of about 150 rpm.

Stage b) - Heating the first mixture to the temperature of the first heating to obtain the second mixture

In some embodiments of the present invention, the first heating temperature is from about 20°C to about 85°C. In some embodiments, the first heating temperature is from about 20°C to about 83°C. In some embodiments, the first heating temperature is from about 20°C to about 55°C. In some embodiments, the first heating temperature is from about 25°C to about 45°C. In some embodiments, the first heating temperature is from about 50°C to about 83°C. In some embodiments, the first heating temperature is from about 60°C to about 80°C. In some embodiments, the first heating temperature is from about 70°C to about 80°C. In some embodiments, the first heating temperature is from about 79°C to about 85°C.

In some embodiments, additional H ₂ O is optionally added to the second mixture.

In some embodiments of the present invention, when water is added to the second mixture, the weight of the water added is from about 0.4 to about 2.3 times the weight of L-arginine originally added when preparing the first mixture.

In some embodiments of the present invention, when water is added to the second mixture, the weight ratio of the water added to the L-arginine originally added in preparing the first mixture is from about 1.0:1.6 to about 1.0:2.1. In some embodiments, when water is added to the second mixture, the weight ratio of the water added to the L-Arginine originally added in preparing the first mixture is from about 1.0:0.8 to about 1.0:1.9. In some embodiments, when water is added to the second mixture, the weight ratio of the water added to the L-arginine initially added to form the first mixture is from about 1.0:1.0 to about 1.0:1.7. In some embodiments, when water is added to the second mixture, the weight ratio of the water added to the L-arginine originally added in preparing the first mixture is from about 1.0:1.1 to about 1.0:1.6. In some embodiments, when water is added to the second mixture, the weight ratio of the water added to the L-arginine originally added in preparing the first mixture is from about 1.0:1.2 to about 1.0:1.5.

In some embodiments of the present invention, the second mixture is essentially a homogeneous solution.

In some embodiments of the present invention, step b) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, step b) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step b) is carried out at a mixing speed of approximately 150 rpm.

Step c) - Adding a first additional amount of water-miscible antisolvent to the second mixture while maintaining the first heating temperature to obtain a suspension

In some embodiments of the present invention, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:5.46 to about 1.00:8.20.

In some embodiments of the present invention, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:4.70 to about 1.00:7.50. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid and the first additional amount of 2-propanol is from about 1.00:5.20 to about 1.00:7.00. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid and the first additional amount of 2-propanol is from about 1.00:5.70 to about 1.00:6.50. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid and the first additional amount of 2-propanol is from at

- 33 045975 approximately 1.00:5.90 to approximately 1.00:6.30. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid and the first additional amount of 2-propanol is from about 1.00:5.95 to about 1.00:6.25. In some embodiments, the weight ratio of L-arginine salt of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B]indol-3-yl)acetic acid and the first additional amount of 2-propanol is from about 1.00:6.00 to about 1.00:6.20.

In some embodiments of the present invention, the weight ratio of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl) acetic acid and the first additional amount of 2-propanol is from about 1.00:5.46 to about 1.00:8.20, from about 1.00:5.81 to about 1.00:7.86, from about 1.00:6.15 to about 1.00:7.51, from about 1.00:6.28 to about 1.00:7.38, from about 1.00:6.42 to about 1.00 :7.24, from about 1.00:6.56 to about 1.00:7.10, from about 1.00:6.69 to about 1.00:6.97 or about 1.00:6, 83.

In some embodiments of the present invention, a first additional amount of water-miscible antisolvent is added at a first time point and at a second time point.

In some embodiments of the present invention, from about 5% to about 15% of the first additional amount of water-miscible antisolvent is added at a first time point. In some embodiments, from about 7% to about 13% of the first additional amount of water-miscible antisolvent is added at the first time point. In some embodiments, from about 8% to about 12% of the first additional amount of water-miscible antisolvent is added at a first time point. In some embodiments, from about 9% to about 11% of the first additional amount of water-miscible antisolvent is added at the first time point.

In some embodiments of the present invention, a first additional amount of water-miscible antisolvent is added at a first time to produce a cloudy mixture.

In some embodiments of the present invention, a seed crystal of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] is optionally added before the second time point indol-3-yl)acetic acid.

In some embodiments of the present invention, a first additional amount of water miscible antisolvent is added at a second time point at a rate such that the addition is completed in about 1.00 hour or more. In some embodiments, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed within about 1.00 hours to about 6.00 hours. In some embodiments, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed within about 1.00 hours to about 5.00 hours. In some embodiments, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed within about 1.00 hours to about 4.00 hours. In some embodiments, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed within about 1.00 hours to about 3.00 hours. In some embodiments, a first additional amount of water-miscible antisolvent is added at a second time point at a rate such that the addition is completed within about 1.00 hours to about 2.00 hours.

In some embodiments of the present invention, the first heating temperature is maintained while the first additional amount of 2-propanol is added to the second mixture.

In some embodiments, step c) is carried out at a mixing speed of from about 100 rpm to about 200 rpm. In some embodiments, step c) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step c) is carried out at a mixing speed of approximately 150 rpm.

Stage d) - Cooling the suspension to the first cooling temperature and subsequent heating to the second heating temperature

In some embodiments of the present invention, cooling the slurry to the first cooling temperature in step d) is carried out at a rate of from about 8.80°Ah to about K^^/hour. In some embodiments, cooling the slurry to the first cooling temperature in step d) is carried out at a rate of from about 9.35°Ah to about 13.80°Ah. In some embodiments, cooling the slurry to the first cooling temperature in step d) is carried out at a rate of from about 9.90°Ah to about 13.20°Ah. In some variants

- 34 045975 cooling of the suspension to the first cooling temperature in step d) is carried out at a rate of from about 10.45°C/hour to about 12.60°C/hour. In some embodiments, cooling the slurry to the first cooling temperature in step d) is carried out at a rate of from about 10°C/hour to about 12°C/hour. In some embodiments, cooling the slurry to the first cooling temperature in step d) is carried out at a rate of from about 9°C/hour to about 11°C/hour. In some embodiments, cooling the slurry to the first cooling temperature in step d) is carried out at a rate of from about 9.5°C/hour to about 10.5°C/hour.

In some embodiments of the present invention, the first cooling temperature in step d) is from about 15°C to about 40°C. In some embodiments, the first cooling temperature in step d) is from about 20°C to about 30°C. In some embodiments, the first cooling temperature in step d) is from about 22°C to about 24°C. In some embodiments, the first cooling temperature in step d) is from about 18°C to about 22°C.

In some embodiments of the present invention, the first cooling temperature is maintained for at least 1 hour before heating to the second heating temperature.

In some embodiments of the present invention, the second heating temperature in step d) is from about 65°C to about 83°C. In some embodiments, the second heating temperature in step d) is from about 70°C to about 80°C. In some embodiments, the second heating temperature in step d) is from about 70°C to about 75°C. In some embodiments, the second heating temperature in step d) is from about 69°C to about 73°C.

In some embodiments of the present invention, after heating to the second heating temperature, the second heating temperature is maintained for at least 30 minutes.

In some embodiments of the present invention, step d) is carried out at a mixing speed of from about 100 revolutions per minute to about 200 revolutions per minute. In some embodiments, step d) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step d) is carried out at a mixing speed of approximately 150 rpm.

Stage e) - Cyclic repetition of stage d) optionally one or more times

It should be understood that when cyclically repeating step d) more than once, the first cooling temperature of each cycle may be the same or different, and the second heating temperature of each cycle may be the same or different.

Two cycles

In some embodiments of the present invention, cycling step e) involves cycling step d) twice.

In some embodiments of the present invention, cycling step d) twice comprises cooling the slurry to the first cycle cooling temperature, heating the slurry to the first cycle heating temperature, cooling the slurry to the second cycle cooling temperature, and heating the suspension to the second cycle heating temperature.

In some embodiments of the present invention, the first cycle cooling temperature is from about 16°C to about 26°C, the first cycle heating temperature is from about 55°C to about 65°C, and the second cycle cooling temperature is from about 26°C to about 36°C, and the heating temperature of the second cycle is from about 45°C to about 55°C. In some embodiments, the first cycle cooling temperature is from about 19°C to about 23°C, the first cycle heating temperature is from about 58°C to about 62°C, and the second cycle cooling temperature is from about 29°C to about 33°C. C, and the heating temperature of the second cycle is from about 48°C to about 52°C.

In some embodiments of the present invention, cooling to the cooling temperature of each cycle is conducted at substantially different cooling rates. In some embodiments, cooling to the cooling temperature of each cycle is conducted at a different cooling rate.

In some embodiments of the present invention, cycling step e) involves cycling step d) twice. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 10.0°C/hour to about 15.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 11.0°C/hour to about 14.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 12.0°C/hour to about 13.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of about 12.5°C/hour.

In some embodiments of the present invention, cycling step e) involves cycling step d) twice. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 7.5°C/hour to about

- 35 045975

12.5°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 8.5°C/hour to about 11.5°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 9.5°C/hour to about 10.5°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of about 10.0° C./hour.

Three cycles

In some embodiments of the present invention, cycling step e) includes cycling step d) three times.

In some embodiments of the present invention, cycling step d) three times comprises cooling the slurry to the first cycle cooling temperature, heating the slurry to the first cycle heating temperature, cooling the suspension to the second cycle cooling temperature, heating the suspension to the second cycle heating temperature, cooling the suspension to the second cycle heating temperature cooling the third cycle and heating the suspension to the heating temperature of the third cycle.

In some embodiments of the present invention, the first cycle cooling temperature is from about 16°C to about 26°C, the first cycle heating temperature is from about 66°C to about 76°C, and the second cycle cooling temperature is from about 16°C to about 26°C, the second cycle heating temperature is from about 55°C to about 65°C, the third cycle cooling temperature is from about 26°C to about 36°C, and the third cycle heating temperature is from about 45°C to about 55 °C. In some embodiments, the first cycle cooling temperature is from about 19°C to about 23°C, the first cycle heating temperature is from about 69°C to about 73°C, the second cycle cooling temperature is from about 19°C to about 23°C, the second cycle heating temperature is from about 58°C to about 62°C, the third cycle cooling temperature is from about 29°C to about 33°C, and the third cycle heating temperature is from about 48°C to about 52°C.

In some embodiments of the present invention, cooling to the cooling temperature of each cycle is conducted at substantially different cooling rates. In some embodiments, cooling to the cooling temperature of each cycle is conducted at a different cooling rate.

In some embodiments of the present invention, cooling to the first cycle cooling temperature and cooling to the second cycle cooling temperature are performed at substantially the same cooling rate.

In some embodiments of the present invention, cycling step e) includes cycling step d) three times. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 10.0°C/hour to about 15.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 11.0°C/hour to about 14.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 12.0°C/hour to about 13.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of about 12.5°C/hour.

In some embodiments of the present invention, cycling step e) includes cycling step d) three times. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 10.0°C/hour to about 15.0°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 11.0°C/hour to about 14.0°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 12.0°C/hour to about 13.0°C/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of about 12.5°C/hour.

In some embodiments of the present invention, cycling step e) includes cycling step d) three times. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 7.5°C/hour to about 12.5°C/hour. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 8.5°C/hour to about 11.5°C/hour. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 9.5°C/hour to about 10.5°C/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of about 10.0° C./hour.

Four cycles

In some embodiments of the present invention, cycling step e) includes cycling step d) four times.

In some embodiments of the present invention, cycling step d) four times includes cooling the slurry to the cooling temperature of the first cycle, heating the suspension to the heating temperature of the first cycle, cooling the suspension to the cooling temperature of the second

- 36 045975 cycle, heating the suspension to the heating temperature of the second cycle, cooling the suspension to the cooling temperature of the third cycle, heating the suspension to the heating temperature of the third cycle, cooling the suspension to the fourth cycle temperature and heating the suspension to the heating temperature of the fourth cycle.

In some embodiments of the present invention, the first cycle cooling temperature is from about 16°C to about 26°C, the first cycle heating temperature is from about 66°C to about 76°C, and the second cycle cooling temperature is from about 16°C to about 26°C, second cycle heating temperature is from about 66°C to about 76°C, third cycle cooling temperature is from about 16°C to about 26°C, third cycle heating temperature is from about 55°C to about 65°C C, the fourth cycle cooling temperature is from about 26°C to about 36°C, the fourth cycle heating temperature is from about 45°C to about 55°C. In some embodiments, the first cycle cooling temperature is from about 19°C to about 23°C, the first cycle heating temperature is from about 69°C to about 73°C, and the second cycle cooling temperature is from about 19°C to about 23°C. C, the second cycle heating temperature is from about 69°C to about 73°C, the third cycle cooling temperature is from about 19°C to about 23°C, the third cycle heating temperature is from about 58°C to about 62°C, the fourth cycle cooling temperature is from about 29°C to about 33°C, the fourth cycle heating temperature is from about 48°C to about 52°C.

In some embodiments of the present invention, cooling to the cooling temperature of each cycle is conducted at substantially different cooling rates. In some embodiments, cooling to the cooling temperature of each cycle is conducted at a different cooling rate.

In some embodiments of the present invention, cooling to the first cycle cooling temperature, to the second cycle cooling temperature, and to the third cycle cooling temperature is carried out at substantially the same cooling rate.

In some embodiments of the present invention, cycling step e) involves cycling step d) four times. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 10.0 Yu/hour to about 15.0 Yu/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of from about IQI/hour to about 14.0 IU/hour. In some embodiments, cooling to the first cycle cooling temperature is conducted at a rate of from about 12.0 Yu/hour to about 13.0 Yu/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of approximately 12.5 Yu/hour.

In some embodiments of the present invention, cycling step e) involves cycling step d) four times. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 10.0 Yu/hr to about 15.0 Yu/hr. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 11.0°O/hour to about 14.0°O/hour. In some embodiments, cooling to the second cycle cooling temperature is conducted at a rate of from about 12.0 Yu/hour to about 13.0 Yu/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of approximately 12.5 Yu/hour.

In some embodiments of the present invention, cycling step e) involves cycling step d) four times. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 10.0 Yu/hour to about 15.0 Yu/hour. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 100,000 units/hour to about 14.0 10,000 units per hour. In some embodiments, cooling to the third cycle cooling temperature is conducted at a rate of from about 12.0 Yu/hour to about 13.0 Yu/hour. In some embodiments, cooling to the first cycle cooling temperature is carried out at a rate of approximately 12.5 Yu/hour.

In some embodiments of the present invention, cycling step e) involves cycling step d) four times. In some embodiments, cooling to the fourth cycle cooling temperature is conducted at a rate of from about 7.5 Yu/hr to about 12.5 Yu/hr. In some embodiments, cooling to the fourth cycle cooling temperature is conducted at a rate of from about 8.5 Yu/hour to about PDY/hour. In some embodiments, the fourth cycle cooling temperature is cooled at a rate of from about 9.5 Yu/hr to about 10.5 Yu/hr. In some embodiments, cooling to the second cycle cooling temperature is carried out at a rate of about 10.0 Yu/hour.

In some embodiments of the present invention, step e) is carried out at a mixing speed of from about 100 revolutions per minute to about 200 revolutions per minute. In some embodiments, step e) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step e) is carried out at a mixing speed of approximately 150 rpm.

- 37 045975

Step f) - Cooling the suspension to the final cooling temperature to obtain the free-plate crystalline form of L-arginine salt (D)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid

In some embodiments of the present invention, the cooling of the suspension in step f) is carried out at a rate of from about 7.5°C/hour to about PD^/hour. In some embodiments, the cooling of the slurry in step f) is carried out at a rate of from about 8.5°C/hour to about PD^/hour. In some embodiments, the cooling of the suspension in step f) is carried out at a rate of from about 9.5°C/hour to about 10°C/hour. In some embodiments, the cooling of the suspension in step f) is carried out at a rate of approximately 100 m/hour.

In some embodiments of the present invention, the cooling of the suspension in step f) is carried out at a rate of from about 8.8°C/hour to about MD^/hour. In some embodiments, the cooling of the slurry in step f) is carried out at a rate of from about 9.4°C/hour to about 13.8°Ο/431Ο. In some embodiments, the cooling of the slurry in step f) is carried out at a rate of from about 9.9°C/hour to about 13.2°C/hour. In some embodiments, the cooling of the slurry in step f) is carried out at a rate of from about 10.5°C/hour to about PD^/hour. In some embodiments, the cooling of the suspension in step f) is carried out at a rate of from about 10°C/hour to about 12°C/hour.

In some embodiments of the present invention, after cooling in step f), the temperature of the slurry is from about 15°C to about 40°C. In some embodiments, after cooling in step f), the temperature of the slurry is from about 20°C to about 30°C. In some embodiments, after cooling in step f), the temperature of the slurry is from about 22°C to about 24°C. In some embodiments, after cooling in step f), the temperature of said suspension is from about 18°C to about 22°C.

In some embodiments of the present invention, step f) is carried out at a mixing speed of from about 100 revolutions per minute to about 200 revolutions per minute. In some embodiments, step f) is carried out at a mixing speed of from about 125 rpm to about 175 rpm. In some embodiments, step f) is carried out at a mixing speed of approximately 150 rpm.

In some embodiments of the present invention, the method further includes the step of isolating the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid.

In some embodiments of the present invention, the isolation step comprises filtering the free-plate crystalline form of L-arginine salt (D)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid from the suspension.

In some embodiments, the isolation step includes the step of filtering the free-plate crystalline form of L-arginine salt (D)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b ]indol-3-yl)acetic acid from the suspension and drying the free-plate crystalline form of L-arginine salt under reduced pressure.

In some embodiments of the present invention, after filtration, the free-plate crystalline form of L-arginine salt (D)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid has one or more properties that are described in the present invention.

Another aspect of the present invention relates to pharmaceutical compositions prepared in accordance with any of the methods described in the present invention.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid obtained in accordance with any of the methods described in the present invention.

The composition contains a free-plate crystalline form of L-arginine salt (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid obtained in accordance with any of the methods described in the present invention.

The pharmaceutical composition contains a free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indol-3-yl) acetic acid obtained in accordance with any of the methods described in the present invention, and a pharmaceutical excipient.

In some embodiments of the present invention, the pharmaceutical composition is suitable for oral, rectal, nasal, topical, buccal, sublingual, vaginal administration, or is in a form suitable for administration by inhalation, insufflation, or transdermal patch. In some embodiments, the pharmaceutical composition is suitable for oral administration.

Lamellar crystal form or morphology

- 38 045975

The present invention relates in particular to the free-plate crystal form or morphology and methods useful for preparing the free-plate crystal form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy) -1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid. The plates have been prepared by the new synthesis methods described in the present invention and have been shown to be thin plates of a hexagon-like shape (hexagonal shape) in which two opposite sides are longer than the other sides (i.e., elongated hexagonal plates) . However, due to the thinness of the plates, an intact plate is very rare. Instead, fragments of thin hexagonal plates of large to small sizes are usually observed. In fig. 5B is a copy of the image shown in FIG. 5A, in which a contour is added defining a substantially intact/intact free plate of an elongated hexagonal shape. One skilled in the art will appreciate that microscopy is one of the most suitable techniques to distinguish between two crystalline forms (habitus) or morphologies. This is particularly applicable when 2 or more morphologies are associated with the same or substantially the same crystalline phase, as is the case with the L-arginine salt of compound 1. Comparison of PXRD X-ray patterns of samples of the crystalline form previously obtained (i.e. in WO2011/094008 and Example 2 below) and the platelet crystal form obtained in accordance with the present invention (ie, see Fig. 19, overlay of PXRD X-ray patterns of spherulites and platelets) shows that both PXRD X-ray patterns are the same or essentially the same, therefore both of these forms represent the same crystalline phase. Although these forms produce the same or substantially the same PXRD patterns, the platelet form exhibits a higher degree of crystallinity, as seen in FIG. 19 essentially due to higher peak intensities as well as lower background scattering attributed to the sample (i.e., less contribution from amorphous halogen). Because the size of the sample and the method of preparation can affect the peak intensities and background scattering associated with the sample, and because the two forms represent the same crystalline phase, PXRD cannot be considered the most suitable method for distinguishing between the two forms. However, PXRD can determine whether crystalline forms belong to the same or different crystalline phases. To determine different crystalline forms, one of the most suitable methods is microscopy. Accordingly, one skilled in the art will be able to examine micrographs of a crystalline form produced by the methods of the present invention and recognize that the crystalline form is a free plate or pieces resulting from fracture of a free plate, and not some other crystalline form, e.g. form of the previous level. Likewise, by simply examining two microphotographs, such as the microphotographs shown in FIG. 1 (spherulites) and Fig. 5A (free plates), it can be easily seen that they are different crystal forms.

In addition to the methods described in the present invention, specific surface area can also be used to characterize the free plates of the present invention. Accordingly, the specific values of the specific surface areas presented in the present invention were obtained using a technique based on the BET (Brunauer, Emmett and Teller) theory, which is a generally accepted theory known in the art for calculating the surface area of solids by measuring the physical adsorption thereof gas molecules (see Brunauer, S.; Emmett, P.H.; and Teller, E.; J. Am. Chem. Soc., 1938, 60, 309). In particular, the specific specific surface area values determined in the present invention were calculated from the BET specific surface area plot obtained by determining the number of nitrogen gas molecules adsorbed by a suspended amount of solid at various relative pressures (P/Po) in the range of 0. 05-0.3 (P/Po) at 77.3 K (-195.85°C). Quantitative determination of the number of adsorbed gas molecules was carried out using a MicromeriticsTM TriStar II BET surface analyzer, the characteristics of which are described below in Example 14. Specifically, nitrogen gas was used to vary the adsorption. The sample for each analysis was degassed under vacuum (100 mm/Hg) at 25°C for 960 minutes. Quantitative determination of nitrogen adsorption was carried out at 77.3 K (195.85 °C) at eleven relative pressures (P/Po), sufficiently dispersed in the range of about 0.05 to about 0.30 (i.e., eleven absolute pressures ranging from about 36 mmHg to about 223 mmHg saturation pressure during measurement, which ranges from about 738 mmHg to about 743 mmHg).

Another aspect of the present invention relates to a new platelet crystal morphology of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid.

In some embodiments of the present invention, the X-ray powder diffraction pattern

- 39 045975 free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2° and 20.5°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 20.5°±0.2°, and 24.6°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2°, and 24. 6°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2°, 24. 6°±0.2° and 28.8°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2°, 24.6°±0.2°, 28.8°±0.2° and 37.3°±0.2°.

In some embodiments of the present invention, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic onset temperature in the range of 205.0°C to 208.5°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 205.0°C to 208.1°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 205.5°C to 208.5°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystalline form has an endotherm with an extrapolated endothermic transition temperature in the range of 206.0°C/hour to 208.5°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 206.5°C to 208.5°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 205.5°C to 208.1°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 206.5°C to 208.1°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 207.0°C to 208.1°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of a free-plate crystal form obtained by scanning at a rate of 10° C./min has an endotherm, as substantially represented in any of Figures 6 through 10. In some embodiments, the differential scanning calorimetry curve of Scanning calorimetry of the free-plate crystalline form obtained by scanning at a rate of 10°C/min has an endotherm as substantially represented in any of Figures 6 to 10 and 22 to 25.

In some embodiments of the present invention, a dynamic moisture sorption (DMS) plot of a free-lamellar crystalline form with an adsorption phase from 30% RH to 90% RH shows an increase in the weight of the free-plate crystalline form of about 0.3% by weight or less at 90% R.H. In some embodiments, a dynamic moisture sorption (DMS) plot of a free-lamellar crystalline form with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-plate crystalline form of about 0.2 wt.% or less at 90% RH.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid with a BET specific surface area of about 0.05 m ² /g, about 0.1 m ² /g, about 0.15 m ² /g, about 0.2 m ² /g , about 0.25 m/g, about 0.3 m ² /g, about 0.35 m/g, about 0.4 m ² /g, about 0.45 m ² /g, about 0.5 m ² /g, about 0.55 m ² /g, about 0.6 m ² /g, about 0.65 m ² /g or from about 0.7 m ² /g to about 2.0 m ² /g, about 2.5 m ² /g, about 3.0 m ² /g, about 3.5 m ² /g, about 4.0 m ² /g, about 4.5 m ² /g, about 5.0 m ² /g, about 5.5 m ² /g, about 6.0 m ² /g, about 6.5 m ² /g, about 7.0 m ² /g, about 7.5 m ² /g, about 8 .0 m ² /g, about 8.5 m ² /g, about 9.0 m ² /g or about 9.5 m ² /g.

In some embodiments of the present invention, the BET specific surface area of the free-plate crystalline form is from about 0.1 m ² /g to about 5.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.1 m ² /g to about 4.0 m ² /g. Somehow

- 40 045975 In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.3 m ² /g to about 4.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.5 m ² /g to about 4.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.6 m ² /g to about 4.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.3 m ² /g to about 3.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.4 m ² /g to about 2.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.5 m ² /g to about 1.8 m ² /g. In some embodiments, the BET specific surface area of the free-lamellar crystalline form is from about 0.6 m ² /g to about 1.6 m ² /g.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) the powder X-ray diffraction pattern has peaks, in units of 2θ, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2°, 24.6°±0 ,2°, 28.8°±0.2° and 37.3°±0.2°;

2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.0°C to 208.5°C when scanning at a speed of 10°C; and/or

3) A plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.3% by mass or less at 90% RH.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) the X-ray powder diffraction pattern has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° and 24.6°±0 ,2°;

2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 206.5°C to 208.5°C when scanning at a speed of 10°C/min; and/or

3) A plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.3% by mass or less at 90% RH.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) the X-ray powder diffraction pattern has peaks, in 2θ units, at 8.2°±0.2°, 20.5°±0.2° and 24.6°±0.2°;

2) the differential scanning calorimetry curve has an endotherm with an extrapolated onset temperature of 205.5°C to 208.5°C when scanning at a speed of 10°C/min; and/or

3) A plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.2% by mass or less at 90% RH.

In some embodiments of the present invention, the free-plate crystal form is:

1) the X-ray powder diffraction pattern has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2° and 20.5°±0.2°;

2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 207.1°C to 208.1°C when scanning at a speed of 10°C/min; and/or

3) A plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.2% by mass or less at 90% RH.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.0°C to 208.5°C when scanning at a speed of 10°C/min; and/or

2) A plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.3% by mass or less at 90% RH.

In some embodiments of the present invention, the free-plate crystalline form has the following properties:

1) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 206.5°C to 208.5°C when scanning at a speed of 10°C/min; and/or

2) A plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.3% by mass or less at 90% RH.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.5°C to 208.5°C when scanning at a speed of 10°C/min; and/or

2) A plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.2% by mass or less at 90% RH.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 207.1°C to 208.1°C when scanning at a speed of 10°C/min; and/or

2) A plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.2% by mass or less at 90% RH.

In some embodiments, the free-lamellar crystalline form has the following properties:

1) the X-ray powder diffraction pattern has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2° and 20.5°±0.2°;

2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.0°C to 208.5°C when scanning at a speed of 10°C/min;

3) a plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.3% by mass or less at 90% RH; and/or

4) BET specific surface area ranges from about 0.1 m ² /g to about 5.0 m ² /g.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) the X-ray powder diffraction pattern has peaks, in 2θ units, at 8.2°±0.2°, 20.5°±0.2° and 24.6°±0.2°;

2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.5°C to 208.5°C when scanning at a speed of 10°C/min;

3) a plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.3% by mass or less at 90% RH; and/or

4) BET specific surface area ranges from about 0.1 m ² /g to about 4.0 m ² /g.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) the X-ray powder diffraction pattern has peaks, in 2θ units, at 8.2°±0.2°, 20.5°±0.2° and 24.6°±0.2°;

2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.5°C to 208.5°C when scanning at a speed of 10°C/min;

3) a plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.3% by mass or less at 90% RH; and/or

4) BET specific surface area ranges from about 0.3 m ² /g to about 3.0 m ² /g.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) the X-ray powder diffraction pattern has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° and 24.6°±0 ,2°;

2) on the differential scanning calorimetry curve there is an endotherm with extrapolation

- 42 045975 bath temperature of the beginning of the endothermic transition in the region from 205.5°C to 208.5°C when scanning at a speed of 10°C/min;

3) a plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.3% by mass or less at 90% RH; and/or

4) specific surface area according to the BET method, ranges from about 0.6 m ² /g to about 4.0 m ² /g.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) the X-ray powder diffraction pattern has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° and 24.6°±0 ,2°;

2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 206.5°C to 208.5°C when scanning at a speed of 10°C/min;

3) a plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.3% by mass or less at 90% RH; and/or

4) specific surface area according to the BET method, ranges from about 0.4 m ² /g to about 2.0 m ² /g.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) the powder X-ray diffraction pattern has peaks, in units of 2θ, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2°, 24.6°±0 .2° and 28.8°±0.2°;

2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 206.5°C to 208.1°C when scanning at a speed of 10°C/min;

3) a plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.2% by mass or less at 90% RH; and/or

4) specific surface area according to the BET method, ranges from about 0.5 m ² /g to about 1.8 m ² /g.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) the powder X-ray diffraction pattern has peaks, in units of 2θ, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2°, 24.6°±0 ,2°, 28.8°±0.2° and 37.3°±0.2°;

2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.5°C to 208.1°C when scanning at a speed of 10°C/min;

3) a plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.2% by mass or less at 90% RH; and/or

4) BET specific surface area ranges from about 0.6 m ² /g to about 4.0 m ² /g.

In some embodiments of the present invention, the free-lamellar crystalline form has the following properties:

1) the powder X-ray diffraction pattern has peaks, in units of 2θ, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2°, 24.6°±0 ,2°, 28.8°±0.2° and 37.3°±0.2°;

2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 207.1°C to 208.1°C when scanning at a speed of 10°C/min;

3) a plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.2% by mass or less at 90% RH; and/or

4) BET specific surface area ranges from about 0.6 m ² /g to about 1.6 m ² /g.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclic lopenta[b] indol-3-yl)acetic acid, the powder X-ray diffraction pattern of which has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2° and 20.5°±0.2 °. In some embodiments, the X-ray powder diffraction pattern of the free-lamellar crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 20.5°±0.2°, and 24.6°±0.2° . In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° and 24.6°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° , 24.6°+0.2° and 28.8°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-plate cree

- 43 045975 steel form there are peaks, in units of 2θ, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2°, 24.6°±0, 2°, 28.8°±0.2° and 37.3°±0.2°.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, on the differential scanning calorimetry curve of which there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.0°C to 208.5°C when scanning at a speed of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 205.0°C to 208.1°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 205.5°C to 208.5°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 205.5°C to 208.1°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 206.0°C to 208.5°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 206.5°C to 208.5°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 206.5°C to 208.1°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of the free-plate crystal form has an endotherm with an extrapolated endothermic transition temperature in the region of 207.0°C to 208.1°C when scanning at a rate of 10°C/min. In some embodiments, the differential scanning calorimetry curve of a free-plate shape obtained by scanning at a rate of 10° C./min has an endotherm, as substantially represented in any of Figures 6 through 10. In some embodiments, the differential scanning calorimetry curve of the free-plate shape calorimetry of a free-plate shape obtained by scanning at a speed of 10° C./min., there is an endotherm, as essentially represented in any of figures 6 to 10 and 22 to 25. In some embodiments of the present invention, the X-ray powder diffraction pattern is free -plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2° and 20.5°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-lamellar crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 20.5°±0.2°, and 24.6°±0.2° . In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° and 24.6°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° , 24.6°±0.2° and 28.8°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° , 24.6°±0.2°, 28.8°±0.2° and 37.3°±0.2°.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, the dynamic moisture sorption (DMS) plot of which with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of about 0.3% by mass or less at 90% RH . In some embodiments of the present invention, a dynamic moisture sorption (DMS) plot of a free-plate crystal form with an adsorption phase from 30% RH to 90% RH shows an increase in the weight of the free-plate crystal form by about 0.2 wt%. or less at 90% RH. In some embodiments, the powder X-ray diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, and 20.5°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-lamellar crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 20.5°±0.2°, and 24.6°±0.2° . In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° and 24.6°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°,

- 44 045975

16.4°±0.2°, 20.5°±0.2°, 24.6°±0.2° and 28.8°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-lamellar crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°+0.2° , 24.6°±0.2°, 28.8°±0.2° and 37.3±0.2°.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid with a BET specific surface area of from about 0.1 m ² /g to about 5.0 m ² /g. In some embodiments of the present invention, the BET specific surface area of the free-plate crystalline form is from about 0.1 m ² /g to about 4.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.3 m ² /g to about 4.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.5 m ² /g to about 4.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.6 m ² /g to about 4.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.3 m ² /g to about 3.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.4 m ² /g to about 2.0 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.5 m ² /g to about 1.8 m ² /g. In some embodiments, the BET specific surface area of the free-plate crystalline form is from about 0.6 m ² /g to about 1.6 m ² /g. In some embodiments of the present invention, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, and 20.5°±0. 2°. In some embodiments, the X-ray powder diffraction pattern of the free-lamellar crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 20.5°±0.2°, and 24.6°±0.2° . In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° and 24.6°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-plate crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° , 24.6°±0.2° and 28.8°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the free-lamellar crystalline form has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°+0.2° , 24.6°±0.2°, 28.8°±0.2° and 37.3°±0.2°.

Indications

S1P receptor agonists with S1P ₁ receptor agonist activity have been shown to rapidly and reversibly induce lymphopenia (also called peripheral lymphocyte lowering PLL); Hale et al., Bioorg. Med. Chem. Lett., 14 :3351-3355, 2004). This is accompanied by clinically beneficial immunosuppression due to the sequestration of T and B cells in secondary lymphoid tissue (lymph nodes and Peyer's patches) and thus away from sites of inflammation and transplanted organs (Rosen et al., Immunol. Rev., 195:160 -177, 2003; Schwab et al., Nature Immunol., 8:1295-1301, 2007). This sequestration of lymphocytes, for example in lymph nodes, is believed to be a consequence of competitive agonist-driven functional antagonism of the S1P ₁ receptor on T cells (resulting in a reduced ability of S1P to mobilize T cell exit from the lymph nodes) and persistent agonism of the S1P1 receptor towards lymph node endothelium (so that the barrier function against lymphocyte transmigration is increased) ((Matloubian et al., Nature, 427:355-360, 2004; Baumruker et al., Expert Opin. Investig. Drugs, 16:283-289, 2007) It has been reported that agonism of the S1P1 receptor alone is sufficient to achieve lymphocyte sequestration (Sanna et al., J Biol Chem., 279:13839-13848, 2004), and that this occurs without impairing the immune response to systemic infection (Brinkmann et al., Transplantation, 72:764-769, 2001; Brinkmann et al., Transplant Proc, 33:530-531, 2001).

This endothelial S1P1 receptor agonism plays a more important role in improving vascular integrity and is supported by work implicating the S1P1 receptor in capillary integrity in mouse skin and lung (Sanna et al., Nat Chem Biol., 2:434-441, 2006). Vascular integrity can be compromised by inflammatory processes, such as sepsis, major trauma, and surgery, leading to acute lung injury or respiratory distress syndrome (Johan Groeneveld, Vascul. Pharmacol., 39:247-256, 2003).

A typical example of an S1P1 receptor agonist with S1P1 receptor agonist activity is the immunosuppressant FTY720 (Martini et al., Expert Opin. Investig. Drugs, 16:505-518, 2007), which has been approved by the Food and Drug Administration US funds (Food and Drug Administration - FDA) as Gilenya® (Fingolimod) in September 2010 for the treatment of relapsing multiple sclerosis. FTY720 acts as a prodrug that is phosphorylated in vivo; the phosphorylated derivative is

- 45 045975 agonist for S1P|, S1P3, S1P ₄ and S1P5 receptors (but not for S1P2 receptor) (Chiba, Pharmacology & Therapeutics, 108:308-319, 2005). FTY720 has been shown to rapidly and reversibly induce lymphopenia (also called peripheral lymphocyte lowering PLL; Hale et al., Bioorg. Med. Chem. Lett., 14:3351-3355, 2004). Clinically beneficial immunosuppression is observed by removing T and B cells in secondary lymphoid tissue (lymph nodes and Peyer's patches) and thus away from sites of inflammation and transplanted organs (Rosen et al., Immunol. Rev., 195 :160-177, 2003; Schwab et al., Nature Immunol., 8:1295-1301, 2007).

FTY720 has been reported to have therapeutic efficacy in at least a rat model of autoimmune myocarditis and a mouse model of acute viral myocarditis (Kiyabayashi et al., J. Cardiovasc. Pharmacol., 35:410-416, 2000; Miyamoto et al., J. Am Coll. Cardiol., 37:1713-1718, 2001); in mouse models of inflammatory bowel disease, including colitis (Mizushima et al., Inflamm. Bowel Dis., 10:182-192, 2004; Deguchi et al., Oncology Reports, 16:699-703, 2006; Fujii et al., Am. J. Physiol. Gastrointest. Liver Physiol., 291: G267-G274, 2006; Daniel et al., J. Immunol., 178: 2458-2468, 2007); a rat model of progressive mesangial proliferative glomerulonephritis (Martini et al., Am. J. Physiol. Renal Physiol., 292:F1761-F1770, 2007); in a mouse model of asthma, proposed to be primarily via the S1P ₁ receptor, based on work using the S1P ₁ receptor agonist SEW2871 (Idzko et al, J. Clin. Invest., 116:2935-2944, 2006); in a mouse model of airway inflammation and induction of bronchial hyperreactivity (Sawicka et al., J. Immunol., 171; 6206-6214, 2003); a mouse model of atopic dermatitis (Kohno et al. r Biol. Pharm. Bull., 27:1392-1396, 2004); in a mouse model of ischemia-reperfusion injury (Kaudel et al., Transplant. Proc, 39:499-502, 2007); in a mouse model of systemic lupus erythematosus (SLE) (Okazaki et al., J. Rheumatol., 29:707-716, 2002; Herzinger et al., Am. J. Clin. Dermatol., 8:329-336 , 2007); in a rat model of rheumatoid arthritis (Matsuura et al., Int. J. Immunopharmacol., 22:323-331, 2000; Matsuura et al., Inflamm. Res., 49:404-410, 2000); a rat model of autoimmune uveitis (Kurose et al., Exp. Eye Res., 70:7-15, 2000); mouse models of type I diabetes (Fu et al., Transplantation, 73: 1425-1430, 2002; Maki et al., Transplantation, 74:1684-1686, 2002; Yang et al., Clinical Immunology, 107:30-35, 2003 ; Maki et al., Transplantation, 79:10511055, 2005); mouse models of atherosclerosis (Nofer et al., Circulation, 115:501-508, 2007; Keul et al., Arterioscler. Thromb. Vase. Biol., 27:607-613, 2007); rat models of brain inflammatory response after traumatic brain injury (TBI); (Zhang et al., J. Cell. Mol. Med., 11:307-314, 2007); and in mouse models of coronary artery disease and graft-versus-host disease (GVHD) (Hwang et al., Circulation, 100:1322-1329, 1999; Taylor et al., Blood, 110:3480-3488 , 2007). In vitro results indicate that FTY720 may have therapeutic efficacy against β-amyloid-related inflammatory diseases, including Alzheimer's disease (Kaneider et al., FASEB J., 18:309-311, 2004). KRP-203, an S1P receptor agonist with S1P ₁ receptor agonist activity, has been reported to have therapeutic efficacy in a rat model of autoimmune myocarditis (Ogawa et al., BBRC, 361:621-628, 2007). Using the S1P ₁ receptor agonist SEW2871S IPI, endothelial S1P1 receptor agonism was shown to prevent proinflammatory monocyte/endothelial interactions in the vascular endothelium in type I diabetes (Whetzel et al., Circ. Res., 99:731-739, 2006) and protect vasculature from TNFa-mediated monocyte/endothelial interactions (Bolick et al., Arterioscler. Thromb. Vasc. Biol., 25:976-981, 2005).

FTY720 has also been reported to have therapeutic efficacy in experimental autoimmune encephalomyelitis (EAE) in rats and mice, a model of human multiple sclerosis (Brinkmann et al., J. Biol. Chem., 277:21453-21457, 2002; Fujino et al., J. Pharmacol. Exp. Ther., 305:70-77, 2003; Webb et al., J. Neuroimmunol., 153:108-121, 2004; Rausch et al., J. Magn. Reson. Imaging, 20:16-24, 2004; Kataoka et al., Cellular & Molecular Immunology, 2:439-448, 2005; Brinkmann et al., Pharmacology & Therapeutics, 115:84-105, 2007; Baumruker et al., Expert Opin. Investig. Drugs, 16:283-289, 2007; Balatoni et al., Brain Research Bulletin, 74:307-316, 2007). In addition, FTY720 has been found to have therapeutic effects against multiple sclerosis in clinical studies. In a phase II clinical trial for relapsing-remitting multiple sclerosis, FTY720 was found to reduce the number of lesions detected by magnetic resonance imaging (MRI) and clinical disease activity in patients with multiple sclerosis (Kappos et al., N. Engl. J. Med., 355:1124-1140, 2006; Martini et al., Expert Opin. Investig. Drugs, 16:505-518, 2007; Zhang et al., Mini-Reviews in Medicinal Chemistry, 7:845-850, 2007 ; Brinkmann, Pharmacology & Therapeutics, 115:84-105, 2007). FTY720 is currently undergoing phase III clinical trials for activity against relapsing-remitting multiple sclerosis.

FTY720 has also been reported to have antiviral activity. Specific data were obtained from a mouse model of lymphocytic choriomeningitis virus (LCMV), in which mice were infected with Armstrong's LCMV or LCMV clone 13 strain (Premenko-Lanier et al., Nature, 454, 894, 2008).

FTY720 has been reported to attenuate the migration of dendritic cells infected with Francisella tu

- 46 045975 larensis, to the mediastinal lymph node, thus reducing its bacterial colonization. Francisella tularensis is associated with tularemia, glandular ulcer infection, respiratory tract infection, and typhoid disease (E. Bar-Haim et al., PLoS Pathog 4(11): e1000211. doi:10.1371/journal.ppat.1000211, 2008).

Short-term high dose FTY720 has also been reported to rapidly reduce ocular infiltrates in experimental autoimmune uveoretinitis. When administered in the early stages of ocular inflammation, FTY720 rapidly prevents retinal damage. It has also been reported not only to prevent infiltration of target organs, but also to reduce existing infiltration (Raveney et al., Arch. Ophthalmol. 126(10), 1390, 2008).

Treatment with FTY720 has also been reported to attenuate oovariectomy-induced osteoporosis in mice by reducing the number of mature osteoclasts attached to the bone surface. The data presented provide evidence that S1P controls the migratory behavior of osteoclast precursors by dynamically regulating bone mineral homeostasis (Ishii et al., Nature, 458(7237), 524-528, 2009).

S1P ₁ receptor agonism is involved in increasing the survival of oligodendrocyte progenitor cells. The survival of oligodendrocyte progenitor cells is an essential component of the remyelination process. Remyelination of lesions in multiple sclerosis is thought to promote recovery in clinical relapses. (Miron et al., Ann. Neurol., 63:61-71, 2008; Coelho et al., J. Pharmacol. Exp. Ther., 323:626-635, 2007; Dev et al., Pharmacology and Therapeutics, 117:7793, 2008). The S1P1 receptor has also been shown to play an important role in platelet-derived growth factor (PDGF)-induced oligodendrocyte progenitor cell mitogenesis (Jung et al., Glia, 55:1656-1667, 2007).

It has also been reported that S1P1 receptor agonism mediates the migration of neural stem cells towards damaged areas of the central nervous system (CNS), including in a rat model of spinal cord injury ((Kimura et al., Stem Cells, 25:115-124, 2007) .

S1P1 receptor agonism is involved in the inhibition of keratinocyte proliferation (Sauer et al., J. Biol. Chem., 279:38471-38479, 2004; Kim et al., Cell Signal, 16:89-95, 2004). Hyperproliferation of keratinocytes at the entrance to the hair follicle, which is then blocked, and associated inflammation are important pathogenetic factors in acne (Koreck et al., Dermatology, 206:96-105, 2003; Webster, Cutis, 76:4-7, 2005 ).

FTY720 has also been reported to have therapeutic efficacy in inhibiting pathological angiogenesis that may occur during tumor development. The inhibition of angiogenesis by FTY720 is thought to involve its agonism at the S1P1 receptor (Oo et al., J. Biol. Chem., 282;9082-9089, 2007; Schmid et al., J. Cell Biochem., 101:259- 270, 2007). FTY720 has also been reported to have therapeutic efficacy in inhibiting the growth of primary and metastatic tumors in a mouse model of melanoma (LaMontagne et al., Cancer Res., 66:221-231, 2006). In addition, FTY720 has been reported to have therapeutic efficacy in a mouse model of metastatic hepatocellular carcinoma (Lee et al., Clin. Cancer Res., 11:84588466, 2005).

Oral administration of FTY720 to mice has also been reported to potently block VEGF-induced vascular permeability, an important process associated with angiogenesis, inflammation, and pathological conditions such as sepsis, hypoxia, and solid tumor growth (T. Sanchez et al., J. Biol. Chem., 278 (47), 47281-47290, 2003).

FTY720 has been shown to have therapeutic efficacy against transplant rejection both as a monotherapy and in synergistic combination with classical immunosuppressive agents including cyclosporine A, FK506 and RAD (mTOR inhibitor). Unlike the classical immunosuppressive drugs cyclosporine A, FK506 and RAD, FTY720 has been shown to be effective in prolonging graft viability without causing overall immunosuppression, and this difference in action of these drugs is thought to be due to the synergy observed in the combination (Brinkmann et al., Transplant Proc., 33:530-531, 2001; Brinkmann et al., Transplantation, 72:764-769, 2001).

S1P1 receptor agonism has been reported to have therapeutic efficacy in prolonging allograft survival in mouse and rat skin allograft models (Lima et al., Transplant Proc, 36:1015-1017, 2004; Yan et al., Bioorg. & Med. Chem. Lett., 16:3679-3683, 2006). FTY720 has been reported to have therapeutic efficacy in prolonging allograft survival in a rat cardiac allograft model (Suzuki et al., Transpl. Immunol., 4:252-255, 1996). FTY720 has also been reported to act synergistically with cyclosporine A to prolong the survival of rat skin allografts (Yanagawa et al., J. Immunol., 160:5493-5499, 1998), to act synergistically with cyclosporine A and with FK506 to prolong the survival of rat cardiac allografts and acts synergistically with cyclosporine A in prolonging canine kidney allograft survival and monkey kidney allograft survival (Chiba et al., Cell Mol. Biol., 3:11-19, 2006). KRP-203, an S1P receptor agonist, has been reported to have therapeutic efficacy in prolonging allograft survival in a rat skin allograft model and also as a monotherapy

- 47 045975 agent and in synergistic combination with cyclosporine A in a rat heart allograft model (Shimizu et al., Circulation, 111:222-229, 2005). KRP-203 has also been reported to have therapeutic efficacy in combination with mycophenolate mofetil (MMF; a prodrug for which the active metabolite is mycophenolic acid, a purine biosynthesis inhibitor) to prolong allograft survival in both a rat renal allograft model and rat cardiac allograft (Suzuki et al., J. Heart Lung Transplant, 25:302-209, 2006; Fujishiro et al., J. Heart Lung Transplant, 25:825-833, 2006). It has been reported that the S1P receptor agonist AUY954, in combination with a subtherapeutic dose of RAD001 (sertican/everolimus, an mTOR inhibitor), can prolong rat cardiac allograft survival (Pan et al., Chemistry & Biology, 13:12271234, 2006). In a rat small intestinal allograft model, FTY720 acts synergistically with cyclosporine A in prolonging small intestinal allograft survival (Sakagawa et al., Transpl. Immunol., 13:161-168, 2004). FTY720 has also been reported to have therapeutic efficacy in a mouse islet transplant model (Fu et al., Transplantation, 73:1425-1430, 2002; Liu et al., Microsurgery, 27:300-304; 2007), and in studies using human islet cells has been shown to have no adverse effects on human islet cell function (Truong et al., American Journal of Transplantation, 7:2031-2038, 2007).

FTY720 has been reported to reduce nociceptive behavior in a spared nerve injury model of neuropathic pain that is independent of prostaglandin synthesis (O. Costu et al., Journal of Cellular and Molecular Medicine 12(3), 995-1004, 2008).

FTY720 has been reported to attenuate the initiation of contact hypersensitivity (CHS) in mice. Adoptive transfer of immunized murine lymph node cells treated with FTY720 during the sensitization phase was virtually unable to induce a CNS response in recipients (D. Nakashima et al., J. Investigative Dermatology (128(12), 2833-2841, 2008).

It was also reported that prophylactic oral administration of FTY720 (1 mg/kg, three times a week) completely prevented the development of experimental autoimmune myasthenia gravis (EAMG) in C57BL/6 mice (T. Kohono et al., Biological & Pharmaceutical Bulletin , 28(4), 736-739, 2005).

_S1P1 receptor agonists may be used to treat or prevent conditions in which suppression of the immune system or agonism at the S1P1 receptor is indicated, such as lymphocyte-mediated diseases and disorders, transplant rejection, autoimmune diseases and disorders, inflammatory diseases and disorders, and conditions , the cause of which is a violation of the integrity of blood vessels, or conditions associated with angiogenesis, which can be pathological.

In some embodiments of the present invention, the S1P1 receptor-related disorder is selected, for example, from lymphocyte-mediated diseases and disorders, transplant rejection, autoimmune diseases and conditions, inflammatory diseases and disorders (eg, acute and chronic inflammatory conditions), cancer, and conditions which are caused by a violation of the integrity of blood vessels or which are associated with angiogenesis, which can be pathological (which, for example, can occur with inflammation, tumor development and atherosclerosis). Conditions in which immune suppression or S1P1 receptor agonism is indicated include lymphocyte-mediated diseases and disorders, conditions involving compromised vascular integrity, autoimmune diseases and disorders, inflammatory diseases and disorders (eg, acute and chronic inflammatory diseases). conditions), acute or chronic cell rejection, autoimmune liver disease including biliary cirrhosis, primary biliary cirrhosis, autoimmune hepatitis, primary sclerosing cholangitis and primary biliary cholangitis, tissue or solid organ transplants, arthritis including psoriatic arthritis and rheumatoid arthritis, diabetes mellitus diabetes, including type I diabetes and disorders associated with it, myasthenia gravis, demyelinating disease, including multiple sclerosis, ischemia-reperfusion injury, including ischemia-reperfusion injury of the kidneys and heart, inflammatory skin diseases, including psoriasis, atopic dermatitis and acne, hyperproliferative skin diseases, including acne, inflammatory bowel diseases, including Crohn's disease and ulcerative colitis, systemic lupus erythematosus, asthma, uveitis, myocarditis, allergies, atherosclerosis, brain inflammation, including Alzheimer's disease and the inflammatory response of the brain after traumatic brain injury, disease of the central nervous system, including spinal cord injury or cerebral infarction, pathological angiogenesis, including angiogenesis that can occur during primary and metastatic tumor growth, rheumatoid arthritis, diabetic retinopathy and atherosclerosis, cancer, chronic lung diseases, acute lung injury, acute respiratory syndrome, sepsis, etc. In addition, S1P1 receptor agonists are useful for the treatment of microbial and viral infections or diseases.

Another aspect of the present invention relates to a method of treating an S1P1 receptor-related disorder in a patient, comprising administering to the patient in need thereof a therapeutically effective

- 48 045975 effective amount of the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3- yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in the present invention.

Another aspect of the present invention relates to a method of treating an S1P ₁ receptor-related disorder in a patient, comprising administering to the patient in need thereof a therapeutically effective amount of a free-lamellar crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl- 3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in of the present invention, wherein the disorder is selected from the group consisting of a lymphocyte-mediated disease or disorder, an autoimmune disease or disorder, an inflammatory disease or disorder, primary biliary cirrhosis, cancer, psoriasis, psoriatic arthritis, rheumatoid arthritis, Crohn's disease, transplant rejection, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, ulcerative colitis, type I diabetes and acne.

Another aspect of the present invention relates to a method of treating a disorder in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-plate crystalline form of L-arginine salt (D)-2-(7-(4-cyclopentyl-3(trifluoromethyl) )benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in the present invention, wherein the disorder is selected from the group consisting of primary biliary cirrhosis, psoriasis, psoriatic arthritis, rheumatoid arthritis, Crohn's disease, multiple sclerosis, inflammatory bowel disease, systemic lupus erythematosus and ulcerative colitis.

Another aspect of the present invention relates to a method of treating a lymphocyte-mediated disease or disorder in a patient, comprising administering to the patient in need thereof a therapeutically effective amount of a free-lamellar crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl -3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in in the present invention.

Another aspect of the present invention relates to a method of treating primary biliary cirrhosis in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-lamellar crystalline form of L-arginine salt (R)-2-(7(4-cyclopentyl-3 -(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described herein invention,

Another aspect of the present invention relates to a method of treating psoriasis in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl) benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in the present invention.

Another aspect of the present invention relates to a method of treating psoriatic arthritis in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of the free-plate crystalline form of ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy) -1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in the present invention.

Another aspect of the present invention relates to a method of treating rheumatoid arthritis in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl) )benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in the present invention.

Another aspect of the present invention relates to a method of treating Crohn's disease in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl) )benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in the present invention.

Another aspect of the present invention relates to a method of treating multiple sclerosis in patients with

- 49 045975 patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-plate crystalline form of L-arginine salt (K)-2-(7-(4-cyclopentyl-.3(trifluoromethyl)benzyloxy)-1,2 ,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in the present invention.

Another aspect of the present invention relates to a method of treating inflammatory bowel disease in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-plate crystalline form of L-arginine salt (R)-2-(7(4-cyclopentyl-3 - (trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in the present invention .

Another aspect of the present invention relates to a method of treating systemic lupus erythematosus in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-lamellar crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-( trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in the present invention.

Another aspect of the present invention relates to a method of treating ulcerative colitis in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl) )benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described in the present invention.

Another aspect of the present invention relates to a method of treating an autoimmune disease or disorder in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-plate crystalline form of L-arginine salt (R)-2-(7(4-cyclopentyl-3 -(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described herein invention.

Another aspect of the present invention relates to a method of treating an inflammatory disease or disorder in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl- 3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3yl)acetic acid as described in the present invention, a composition as described in the present invention, or a pharmaceutical composition as described herein invention.

Another aspect of the present invention relates to a method of treating a microbial or viral infection or microbial or viral disease in a patient, comprising administering to the patient in need thereof a therapeutically effective amount of a free-lamellar crystalline form of L-arginine salt ^)-2-(7-( 4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol-3-yl)acetic acid which is described in the present invention, a composition which is described in the present invention, or a pharmaceutical composition which described in the present invention.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the manufacture of a medicament for the treatment of S1P ₁ receptor-related disorder.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the manufacture of a medicament for the treatment of an S1P1 receptor-related disorder selected from the group consisting of a disease or disorder mediated by lymphocytes, an autoimmune disease or disorder, an inflammatory disease or disorder, primary biliary liver cirrhosis, cancer, psoriasis, psoriatic arthritis, rheumatoid arthritis, Crohn's disease, transplant rejection, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, ulcerative colitis, type I diabetes and acne.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the manufacture of a medicament for the treatment of an S1P1 receptor-related disorder selected from the group consisting of primary biliary cirrhosis, psoriasis, psoriatic arthritis, rheumatoid arthritis, Crohn's disease, multiple sclerosis, inflammatory bowel disease, systemic lupus erythematosus and ulcerative colitis.

- 50 045975

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the manufacture of a medicament for the treatment of a disease or disorder mediated by lymphocytes.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the production of a medicament for the treatment of primary biliary cirrhosis.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the production of a drug for the treatment of psoriasis.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the production of a drug for the treatment of psoriatic arthritis.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the production of a drug for the treatment of rheumatoid arthritis.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the production of a drug for the treatment of Crohn's disease.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the production of a drug for the treatment of multiple sclerosis.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the production of a drug for the treatment of inflammatory bowel disease.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the production of a drug for the treatment of systemic lupus erythematosus.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the production of a drug for the treatment of ulcerative colitis.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the manufacture of a medicament for the treatment of an autoimmune disease or disorder.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the manufacture of a medicament for the treatment of an inflammatory disease or disorder.

Another aspect of the present invention relates to the use of the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol- 3-yl)acetic acid, which is described in the present invention, in the production of a medicament for the treatment of a microbial or viral infection or a microbial or viral disease.

Another aspect of the present invention relates to the use of a composition containing a free-plate crystalline form of L-arginine salt (K)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid, which is described in the present invention, and an excipient, in the manufacture of a medicament for administration in the treatment of S1P ₁ receptor-related disorder.

Another aspect of the present invention relates to the use of a composition containing a free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid, which is

- 51 045975 sana in the present invention, and an excipient, in the manufacture of a medicament for administration in the treatment of S1P ₁ receptor-related disorder.

Another aspect of the present invention relates to the use of a composition containing a free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid, which is described in the present invention, and an excipient for compounding a drug for administration in the treatment of S1P1 receptor-related disorder.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating a human or animal with a therapy.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating an S1P1 receptor-related disorder.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating S1P ₁ receptor-related selected from the group consisting of a disease or disorder mediated by lymphocytes, an autoimmune disease or disorder, an inflammatory disease or disorder, primary biliary cirrhosis, cancer, psoriasis, psoriatic arthritis, rheumatoid arthritis, Crohn's disease, transplant rejection, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, ulcerative colitis, type I diabetes and acne.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating an S1P ₁ receptor-related disorder selected from the group consisting of primary biliary cirrhosis, psoriasis, psoriatic arthritis, rheumatoid arthritis, Crohn's disease, multiple sclerosis, inflammatory bowel disease, systemic lupus erythematosus and ulcerative colitis.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating a disease or disorder mediated by lymphocytes.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method for treating primary biliary cirrhosis.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method for treating psoriasis.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating psoriatic arthritis.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating rheumatoid arthritis.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating Crohn's disease.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating multiple sclerosis.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, which is described in the present invention, for use

- 52 045975 in a method for treating inflammatory bowel disease.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method for treating systemic lupus erythematosus.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating ulcerative colitis.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating an autoimmune disease or disorder.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating an inflammatory disease or disorder.

Another aspect of the present invention relates to the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[B] indol-3-yl)acetic acid, which is described in the present invention, for use in a method of treating a microbial or viral infection or a microbial or viral disease.

In some embodiments of the present invention, the S1P ₁ receptor-related disorder is mediated by lymphocytes. In some embodiments, the S1P1 receptor-related disorder is primary biliary cirrhosis. In some embodiments, the S1P1 receptor-related disorder is psoriasis. In some embodiments, the S1P ₁ receptor-related disorder is psoriatic arthritis. In some embodiments, the S1P ₁ receptor-related disorder is rheumatoid arthritis. In some embodiments, the S1P ₁ receptor-related disorder is Crohn's disease. In some embodiments, the SLPI receptor-related disorder is multiple sclerosis. In some embodiments, the S1P ₁ receptor-related disorder is an inflammatory bowel disease. In some embodiments, the S1P1 receptor-related disorder is systemic lupus erythematosus. In some embodiments, the S1P1 receptor-related disorder is ulcerative colitis.

Pharmaceutical compositions and formulations

Another aspect of the present invention relates to pharmaceutical compositions containing the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid, which is described in the present invention, and a pharmaceutical excipient.

Another aspect of the present invention relates to pharmaceutical compositions containing the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid obtained in accordance with any of the methods described herein, and a pharmaceutical excipient.

Another aspect of the present invention relates to pharmaceutical compositions containing the free-plate crystalline form of L-arginine salt A)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid (compound 1) in an amount equivalent to a therapeutically effective amount of compound 1 free-plate crystalline form, wherein the pharmaceutical composition further includes a diluent, a disintegrating agent and a lubricant.

In some embodiments of the present invention, the diluent includes a first diluent and a second diluent.

In some embodiments of the present invention, the first diluent includes mannitol. In some embodiments, the first diluent includes mannitol 200 SD, USP.

In some embodiments of the present invention, the second diluent includes microcrystalline cellulose. In some embodiments, the second diluent includes Avicel®. In some embodiments, the second diluent contains Avicel® PH102, NF.

In some embodiments of the present invention, the disintegrant is sodium starch glycolate. In some embodiments, the disintegrant is Explotab®, NF.

In some embodiments of the present invention, the lubricant is

- 53 045975 is magnesium stearate, NF.

In some embodiments of the present invention, the diluent content of the pharmaceutical composition is from about 91% to about 95%.

In some embodiments of the present invention, the content of the first diluent in the pharmaceutical composition is from about 51% to about 55%.

In some embodiments of the present invention, the content of the second diluent in the pharmaceutical composition is from about 37% to about 43%.

In some embodiments of the present invention, the content of the disintegrating agent in the pharmaceutical composition is from about 3% to about 5%.

In some embodiments of the present invention, the lubricant content of the pharmaceutical composition is from about 0.2% to about 0.8%.

In some embodiments of the present invention, the pharmaceutical composition is a pharmaceutical composition comprising the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3 ,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid is 0.69%;

mannitol content is 54.81%;

microcrystalline cellulose content is 40%;

sodium starch glycolate content is 4%; and the magnesium stearate content is 0.5%.

In some embodiments of the present invention, the pharmaceutical composition is a pharmaceutical composition comprising the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3 ,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid is 1.381%;

mannitol content is 54.119%;

microcrystalline cellulose content is 40%;

sodium starch glycolate content is 4%; and the magnesium stearate content is 0.5%.

In some embodiments of the present invention, the pharmaceutical composition is a pharmaceutical composition comprising the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3 ,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid is 2.762%;

mannitol content is 52.73%;

microcrystalline cellulose content is 40%;

sodium starch glycolate content is 4%; and the magnesium stearate content is 0.5%.

In some embodiments of the present invention, the pharmaceutical composition is a pharmaceutical composition wherein the content of the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3, 4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid is 4.143%;

mannitol content is 51.357%;

microcrystalline cellulose content is 40%;

sodium starch glycolate content is 4%; and the magnesium stearate content is 0.5%.

Another aspect of the present invention relates to compositions containing the free-plate crystalline form of L-arginine salt 4)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid, which is described in the present invention or obtained in accordance with any of the methods described in the present invention.

Another aspect of the present invention relates to pharmaceutical compositions containing the free-plate crystalline form of L-arginine salt 4)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[ b]indol-3-yl)acetic acid, which is described in the present description or obtained in accordance with any of the methods described in the present invention, and a pharmaceutical excipient.

In some embodiments of the present invention, the composition and/or pharmaceutical composition is suitable for oral, rectal, nasal, topical, buccal, sublingual, vaginal administration, or is in a form suitable for administration by inhalation, insufflation, or transdermal patch. In some embodiments, the implementation

- 54 045975 item and/or pharmaceutical composition suitable for oral administration.

The preparations can be prepared by any suitable method, usually by uniformly mixing the active compound(s) with liquid or finely divided solid carriers or both in the desired proportions and, if necessary, then molding the resulting mixture into the desired form.

Conventional excipients such as binding agents, fillers, suitable wetting agents, tabletting lubricants and disintegrants may be used in tablets and capsules for oral administration. Liquid preparations for oral administration can be presented in the form of solutions, emulsions, aqueous or oily suspensions and syrups. Alternatively, the oral preparations may be presented in the form of a dry powder which may be reconstituted by the addition of water or other suitable liquid carrier prior to use. Additional ingredients may be added to liquid preparations, such as suspending or emulsifying agents, non-aqueous vehicles (including edible oils), preservatives and flavoring agents and colors. Dosage forms for parenteral administration can be prepared by dissolving a compound of the present invention in a suitable liquid carrier and sterilizing the solution, followed by packaging and sealing in suitable containers or ampoules. These are just a few examples of the many suitable methods for preparing dosage forms well known in the art.

The compounds of the present invention, such as the L-arginine salt of Compound 1, can be formulated into pharmaceutical compositions using methods well known to those skilled in the art. Suitable pharmaceutically acceptable carriers other than those mentioned herein are known in the art; see, for example, The Science and Practice of Pharmacy, ^20th Edition, 2000, Lippincott Williams & Wilkins, (Editors: Gennaro et al.).

The term composition refers to at least one compound of the present invention in combination with at least one other component. Examples of compositions include, but are not limited to, a standard containing a compound of the present invention (eg, for use in method development, process testing, etc.); bulk FAI (ie, pharmaceutically active ingredient) of the compound of the present invention (eg, for preparing a pharmaceutical composition, compounding for preparing a drug for use when administered to a patient, etc.); combination drug (ie, a compound of the present invention in combination with a pharmaceutical/therapeutic drug or pharmaceutical/therapeutic drugs); a biological sample containing a compound of the present invention (e.g., for administration to or from a patient, animal, pharmacokinetic studies, toxic compound absorption, distribution, metabolism, and excretion (ADME) studies, and release, absorption, distribution studies, metabolism, elimination of toxic compounds (LADME studies), etc.); a reaction mixture containing a compound of the present invention, such as the reaction mixture described in any of the examples presented herein; a production reaction mixture containing a compound of the present invention in combination with one or more components such as solvents, reagents, by-products, etc.; and so on. It should be understood that pharmaceutical compositions are a specific subset of compositions. In one embodiment of the present invention, the composition may be used to treat a disease or as described in the present invention.

The compounds of the invention, such as the L-arginine salt of Compound 1, can be combined or used in a compounding process to provide a personalized medicament for a patient or group of patients to treat a disease or disorder as described herein. The techniques used in compounding are well known to those skilled in the art, such as compounding pharmacists. In general, the term compounding refers to the practice of obtaining a drug prescription for a specific patient. Compounding may involve the preparation, mixing, compounding, packaging and/or labeling of a drug product (1) after the drug has been prescribed by a physician or as a result of an independent decision based on the pharmacist/patient/prescriber relationship in professional practice, or (2) for conducting research, teaching, or chemical analysis and not for the sale or dispensing of prescription drugs. Compounding also involves obtaining drugs in advance of filling prescriptions based on common, regularly observed patterns. Compounding services play a vital role in healthcare because commercially produced drugs may not be suitable for every person. Due to patient differences, such as body size, varying drug tolerances, allergies, and/or special requirements (eg, sugar-free, gluten-free, casein-free, soy-free, and/or color-free drug dosing), a patient may require a medication that precise dosage is not commercially available, or the drug may require specific components

- 55 045975 volume, which is not in the commercial preparation. With the help of compounding, the pharmacist can adjust the treatment to the exact dosage of drug and excipient(s) required for the patient, such as prescribing a treatment or drug suitable for oral, rectal, nasal, topical, buccal, sublingual, vaginal, or in the form, suitable for administration by inhalation, insufflation or transdermal patch.

Although, in an alternative prophylactic or therapeutic application, a compound of the present invention may be administered as a crude or pure chemical, it is preferable to provide the compound or active ingredient in the form of a pharmaceutical preparation or composition containing a pharmaceutically acceptable carrier.

Pharmaceutical preparations include preparations suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including intramuscular, subcutaneous and intravenous) administration, or preparations in a form suitable for administration by inhalation, insufflation or transdermal patch . Transdermal patches release drug at a controlled rate, providing the drug for efficient absorption with minimal drug degradation. Typically, transdermal patches include an impermeable backing layer, a pressure-sensitive uniform adhesive layer, and a removable barrier layer with a protective coating. One skilled in the art will understand and appreciate methods suitable for producing a transdermal patch of the required effectiveness based on the manufacturer's requests.

Thus, the compounds of the present invention, such as the L-arginine salt of Compound 1, together with a standard adjuvant, carrier or diluent can be formulated into pharmaceutical preparations and unit dosage forms thereof and in such form can be used as solids such as tablets or filled capsules, or liquids, such as solutions, suspensions, emulsions, elixirs, gels or capsules filled with the said liquids, for oral administration, in the form of suppositories for rectal administration; or in the form of sterile injection solutions for parenteral (including subcutaneous) administration. Such pharmaceutical compositions and unit dosage forms thereof may contain standard ingredients in standard proportions with or without additional active compounds or active ingredients, and such unit dosage forms may contain any suitable effective amount of the active ingredient commensurate with the prescribed daily dosage interval that must be entered.

For oral administration, the pharmaceutical composition may be presented, for example, in the form of a tablet, capsule, suspension or liquid. The pharmaceutical composition is preferably prepared in unit dose form containing a specific amount of the active ingredient. Examples of such unit dosage forms are capsules, tablets, powders, granules or suspensions with conventional additives such as lactose, mannitol, corn starch or potato starch; with binders such as crystalline cellulose, cellulose derivatives, gum arabic, corn starch or gelatins; with disintegrants such as corn starch, potato starch or sodium carboxymethylcellulose; and with lubricants such as talc or magnesium stearate. The active ingredient can also be administered by injection in the form of a composition in which, for example, saline, dextrose or water can be used as a suitable pharmaceutically acceptable carrier.

In some embodiments of the present invention, the pharmaceutical composition is a solid preparation, such as a tablet, capsule, oral suspension, or the like.

The dosage of use of the compounds of the present invention, such as the L-arginine salt of Compound 1, can vary widely and, as is customary and known to the attending physician, should be adapted to the individual condition in each individual case. It depends, for example, on the nature and severity of the disease to be treated, on the condition of the patient, on the compound used, on whether the disease state to be treated is acute or chronic, whether prophylactic treatment has been administered or not, or on the administration of additional compounds in addition to the compounds of the present invention, such as the L-arginine salt of Compound 1. Typical dosages include, but are not limited to, about 0.001 mg to about 5000 mg, about 0.001 mg to about 2500 mg, about 0.001 mg to about 1000 mg, about 0.001 mg to about 500 mg, about 0.001 mg to about 250 mg, about 0.001 mg to 100 mg, about 0.001 mg to about 50 mg, and about 0.001 mg to about 25 mg. Multiple doses may be administered throughout the day, especially when relatively large quantities are needed, such as 2, 3 or 4 doses. Depending on individual characteristics and at the discretion of the attending physician, it may be necessary to increase or decrease the doses described in the present invention.

In some embodiments of the present invention, the dose is 0.5 mg. In some embodiments, the dose is 1.0 mg. In some embodiments, the dose is 2.0 mg. In some embodiments, the dose is 3.0 mg. In some variants

- 56 045975 implementation dose is 5.0 mg.

All doses described in the present invention are based on the active ingredient, that is, the molecule or ion that provides the desired pharmacological or physiological effect.

The amount of active ingredient, active salt or derivative thereof required for use in treatment will vary depending not only on the particular salt selected, but also on the route of administration, the nature of the condition being treated, and the age and condition of the patient, and will ultimately be determined your attending physician or clinician. Typically, one skilled in the art will understand how to extrapolate in vivo data obtained in a model system, typically an animal model, to another model, such as a human. In some cases, this extrapolation may be based simply on a comparison of the mass of the animal and another model, such as a mammal, preferably a human, however, more often than not, this extrapolation is not simply based on body mass, but includes various factors. Typical factors include type, age, body weight, sex, diet and medical condition of the patient, severity of disease, routes of administration, pharmacological properties such as potency, efficacy, pharmacokinetic and toxicological profiles of the particular compound used, use of drug delivery system, acute or chronic is the disease state being treated, whether prophylactic treatment has been performed, and whether other active compounds in addition to the compounds presented herein (eg, L-arginine salt of compound 1) are used as part of the combination preparation. A dosage regimen for treating a disease state with the compounds and/or compositions of the present invention is selected in accordance with the various factors mentioned above. Thus, the actual dosage regimen for a drug may vary widely and therefore deviate from the preferred dosage regimen, and one skilled in the art will appreciate that dosage and dosage regimens of a drug outside of these typical ranges will may be tested and used in the methods described in the present invention if necessary.

The desired dose may be conveniently presented in a single dose or divided into doses administered at suitable intervals, for example into two, three, four or more portions of the daily dose. Part of the daily dose can be further divided, for example, into a number of separate independent administrations. In particular, especially when relatively large amounts are administered, the daily dose may, if necessary, be divided into several administrations, for example 2, 3 or 4 administrations. If necessary, depending on individual behavior, an increase or decrease in the indicated daily dose may be required.

The compounds of the present invention, such as the L-arginine salt of Compound 1, can be administered in various dosage forms for oral and parenteral administration. It will be apparent to one skilled in the art that dosage forms may contain as the active ingredient a compound of the present invention, a pharmaceutically acceptable salt, hydrate or solvate of a compound of the present invention.

To prepare pharmaceutical compositions from the compounds of the present invention, such as the L-arginine salt of Compound 1, the selected suitable pharmaceutically acceptable carrier may be a solid, a liquid, or a mixture thereof. Solid dosage forms include powders, tablets, pills, capsules, wafers, suppositories and dispersible granules. A solid carrier may be one or more substances that may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, disintegrants, or encapsulating material.

In powders, the carrier is a finely divided solid that is mixed with the finely divided active ingredient.

In tablets, the active ingredient is mixed with a carrier having the required binding capacity in suitable proportions and compressed into the desired shape and size.

Powders and tablets may contain varying amounts of the active compound. A typical amount of active compound in a powder or tablet may range from 0.5 to about 90 percent; however, one skilled in the art will recognize when amounts beyond the stated range are needed. Suitable carriers for powders and tablets include magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, etc. The term drug refers to a preparation of the active compound with an encapsulating material as a carrier, providing a capsule in which the active component, with or without the addition of carriers, is surrounded by a carrier, which is thus in association with it. This also applies to wafers and cakes. Tablets, powders, capsules, pills, wafers and lozenges can be used as solid forms suitable for oral administration.

To prepare suppositories, a low-melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted and the active ingredient is homogeneously dispersed therein while stirring

- 57 045975 vaniya. After this, the molten homogeneous mixture is poured into molds of a convenient size, allowed to cool and eventually harden.

Compositions suitable for vaginal administration may be presented in the form of vaginal suppositories, tampons, creams, gels, pastes, foams or sprays containing, when appropriate, in addition to the active ingredient, carriers which are known in the art.

Preparations in liquid form include solutions, suspensions and emulsions, such as aqueous solutions or solutions in a mixture of water and propylene glycol. For example, liquid preparations for parenteral injection may be presented as solutions in an aqueous solution of polyethylene glycol. Injectable preparations, for example sterile aqueous or oleaginous injectable suspensions, can be prepared according to methods known in the art using suitable dispersing, wetting and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or a sterile injectable suspension in a non-toxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol. Acceptable carriers and solvents that may be used are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile non-volatile oils are usually used as a solvent or suspending medium. Any soft fixed oils can be used for this purpose, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of preparations suitable for injection.

Thus, the compounds of the present invention, such as the L-arginine salt of Compound 1, can be formulated for parenteral administration (eg, injection, particularly bolus injection, or continuous infusion) and can be presented in unit dose dosage form at ampoules, pre-filled syringes, in containers containing small volume infusions or multiple doses with the addition of a preservative. Pharmaceutical compositions may take forms such as suspensions, solutions or emulsions in oily or aqueous vehicles and may contain auxiliary additives such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be presented in the form of a powder obtained by aseptic isolation of a sterile solid or lyophilization from solution, for mixing with a suitable vehicle, for example, sterile pyrogen-free water, before use.

Aqueous compositions suitable for oral administration can be prepared by dissolving or suspending the active ingredient in water and adding, if desired, suitable coloring agents, flavoring agents, stabilizers and thickening agents.

Aqueous suspensions suitable for oral administration can be prepared by dispersing the finely divided active ingredient in water containing a viscous material such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose or other well known suspending agents.

Liquid drugs also include solid form drugs that are intended to be converted into a liquid form for oral administration immediately before use. Such liquid forms include solutions, suspensions and emulsions. In addition to the active ingredient, these preparations may contain coloring agents, flavoring agents, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, etc.

For topical administration to the epidermis, the compounds of the present invention, such as the L-arginine salt of Compound 1, can be presented in the form of ointments, creams or lotions or in the form of transdermal patches.

Ointments and creams can be prepared, for example, in an aqueous or oily base with the addition of suitable thickening agents and/or gelling agents. Lotions may be aqueous or oily based and typically also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents or coloring agents.

Formulations suitable for topical administration to the oral cavity include lozenges containing the active agent in a flavored base, usually sucrose and acacia or tragacanth; lozenges containing the active ingredient in an inert base such as gelatin and glycerin or sucrose and gum arabic; and a mouthwash containing the active ingredient in a suitable liquid carrier.

Solutions or suspensions are applied directly into the nasal cavity using conventional means, such as a dropper, pipette or spray. The drugs may be presented in a dosage form containing one or more doses. In the latter case, when using a dropper or pipette, dosing can be achieved by administering a suitable predetermined volume of solution or suspension to the patient. In the case of a sprayer, this can be achieved, for example, using a spray pump with a dispenser.

Administration into the respiratory tract can also be achieved using an aerosol formulation in which the active ingredient is presented in a pressurized package with a suitable propellant.

If the compounds of the present invention, such as the L-arginine salt of Compound 1, or

- 58 045975 pharmaceutical compositions containing them are administered in the form of aerosols, for example in the form of nasal aerosols, or by inhalation, such administration can be carried out, for example, using a spray, nebulizer, pump nebulizer, inhalation device, metered inhaler or dry powder inhaler. Pharmaceutical forms for administering the compounds of the present invention as an aerosol can be prepared by methods well known to one skilled in the art. For their preparation, for example, solutions or dispersions of the compounds of the present invention in water, water/alcohol mixtures or suitable physiological solutions can be used using conventional additives, for example benzyl alcohol or other suitable preservatives, absorption enhancers to increase bioavailability, solubilizers, dispersants, etc. ., and, if necessary, conventional propellants, for example carbon dioxide, chlorofluorocarbons such as dichlorodifluoromethane, trichlorofluoromethane or dichlorotetrafluoroethane, etc. The aerosol may also contain a surfactant such as lecithin. The dose of the drug can be adjusted using a dosing valve.

In preparations intended for administration into the respiratory tract, including preparations for nasal administration, the compound will typically have a small particle size, for example on the order of 10 microns or less. This particle size can be obtained by methods known in the art, for example micronization. If necessary, compositions adapted for sustained release of the active ingredient may be used.

Alternatively, the active ingredients may be presented in dry powder form, for example in the form of a powder mixture of the compound in a suitable powder base such as lactose, starch, starch derivatives such as hydroxypropyl methylcellulose and polyvinylpyrrolidone (PVP). Typically, the powdered carrier forms a gel in the nasal cavity.

The powdered composition may be presented in unit dose dosage form, for example in capsules or cartridges, for example gelatin, or in blister packs from which the powder can be administered by inhaler.

The pharmaceutical preparations are preferably unit dose dosage forms. In this form, the drug is divided into unit doses containing appropriate amounts of the active component. A unit dose dosage form may be a prepackaged preparation whose packaging contains discrete quantities of the drug, such as packaged tablets, capsules and powders in vials or ampoules. In addition, the unit dose dosage form may be a capsule, tablet, cachet, or lozenge, or it may be a package containing an appropriate number of capsules, tablets, wafers, or lozenges.

Tablets or capsules for oral administration and liquids for intravenous administration are preferred compositions.

As will be appreciated, the steps of the methods of the present invention do not have to be performed any particular number of times or in any particular sequence. Additional objects, advantages and novel features of the invention(s) will become apparent to one skilled in the art upon examination of the following examples, which are intended to be illustrative only and not limiting of the present invention.

Examples

The compounds described herein and their synthesis are further illustrated by the following examples. The following examples are provided to further describe the invention, but the invention is not limited to these examples. The compounds described herein above and below are named in accordance with 2.2, CS ChemDraw Ultra Version 9.0.7 or ChemBioDraw Ultra 12.0.2.1076. In some cases, generic names are used, and it will be understood that these generic names will be known to those skilled in the art.

Chemistry: Proton nuclear magnetic resonance spectra ( ^1H NMR) were recorded on a Bruker Avance III400 equipped with a 5 mm BBFO probe. Chemical shifts are given in parts per million (ppm) with the residual solvent signal used as a reference. The following abbreviations are used in NMR spectra: c=singlet, d=doublet, dd=doublet of doublets, t=triplet, q=quartet, m=multiplet, br.s=broadened singlet, sct=sextet. Microwave irradiations were performed using a Smith Synthesizer™ or Emrys Optimizer™ (Biotage). Thin layer chromatography (TLC) was performed on silica gel 60 F254 (Merck), preparative thin layer chromatography (preparative TLC) was performed on silica gel plates PK6F 60 A 1 mm (Whatman), column chromatography was performed on a column with silica gel Kieselgel 60, 0.063-0.200 mm (Merck ). Evaporation was carried out under reduced pressure on a Buchi rotary evaporator. Celite® 545 was used to filter the palladium.

LCMS specifications: HPLC - Agilent 1200; pumps: G1312A; DAD:G1315B; sampler: G1367B; Agilent G1956A mass spectrometer; ionization source: ESI; drying in gas flow: 10 l/min; Spray Pressure: 40 psi (275.390 kPa); drying gas temperature: 350°C; capillary voltage: 2500 V; software: Agilent Chemstation Rev.B.04.03.

Example 1: Preparation and crystallization of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid crystal sphere

- 59 045975 rolitic morphology

A mixture of ethyl 2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta3-yl)acetate (29.0 g, 59.7 mmol, 1.00 eq. ), lipase B (Candida antarctica, immobilized, 1.45 g) and an aqueous solution of potassium phosphate buffer (pH 7.8 ± 0.2, 1.0 M, 21.75 ml) in acetonitrile (268.5 ml) stir under a nitrogen atmosphere at 40°C for at least 8 hours until the product concentration (i.e., ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1.2 ,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid) will not reach >40 area % according to HPLC. The pH value is then adjusted to 4-6 by adding 1.0 M aqueous citric acid solution (26.1 ml) and stirring the mixture at 20°C. Solids are removed by filtration, the reactor and filter cake are washed with acetonitrile (3x10 ml) and the acetonitrile is removed in vacuo, after which the product mixture is partitioned between ethyl acetate (58 ml) and water (58 ml). The ethyl acetate phase is washed with water (2x58 ml), then with brine (3x58 ml) at least twice until the pH of the wash solution changes from 4 to 7, and the citric acid content of the ethyl acetate phase according to 1H NMR analysis -spectroscopy will not be <0.1%. Ethyl acetate is replaced with 2propanol (3x87 ml) using vacuum distillation. After complete removal of ethyl acetate, which was verified by ¹ H NMR spectroscopy, a 2-propanol solution (10 ml/g (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4 -tetrahydrocyclopenta[b]indol-3-yl)acetic acid) product is filtered and analyzed using achiral and chiral HPLC to determine the concentration of (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1 ,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and the corresponding enantiomeric acid (i.e. (S)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1, 2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid). The solution is concentrated to an oil, part of which is used in the preparation of L-arginine salt (R)2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3 -yl)acetic acid as described below.

A mixture of the product in 2-propanol containing approximately equal amounts of ^)-2-(7-(4-cyclopentyl3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid (3.0 g, 6.56 mmol, 1,000 eq.) and the corresponding N-ethyl ether, stir with 2-propanol (50 ml) at 60°C for 5 minutes. Thereafter, L-arginine salt seed material (0.06 g) is added and stirred for 10 minutes. Then, a 2.27 M aqueous solution of L-arginine (2.89 ml, 6.56 mmol, 1.00 eq.) preheated to 60°C was added dropwise over 15 minutes to obtain crystalline L-arginine salt ^)- 2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid. The yellowish suspension is stirred (at approximately 100 rpm) for 60 minutes. The mixture is cooled to 25°C±5°C at a rate of 0.5°C per minute. The L-arginine salt is isolated by filtration and washed with 2-propanol (3x3 ml), then ethyl acetate (3x3 ml) until the impurities in the wet filter cake are less than 1% by area according to HPLC. The product was then dried under vacuum at 40°C ± 5°C for 5 hours to obtain 3.356 g (40.5% yield) of crystalline L-arginine salt in the form of spherulites (Fig. 1 shows a microphotograph of spherulites obtained using a scanning electron microscopy (SEM), Fig. 2 shows a microphotograph of spherulites obtained by optical polarization microscopy (PLV), which shows characteristic crosses during cross-polarization imaging); HPLC: 99.2% by area (achiral), 98.8% w/w. (achiral) and 99.5% by area (chiral); DSC 201.97°C; particle size: average diameter 23.58, 11.46 µm standard deviation, 6.7 µm minimum, 47.38 µm maximum length (ML).

Example 2: Preparation and crystallization of L-arginine salt ^)-2-(7-(4-cyclodopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid

Ethyl 2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]yne dol-3-yl)acetate mixture (21.86 kg, 45.0 mol, 1,000 equiv.), lipase B (Candida antarctica, immobilized 0.64 kg) and 18.4 kg aqueous potassium phosphate buffer (pH 7.8 ± 0.2, 1.0 M) in 161.0 kg acetonitrile stir under a nitrogen atmosphere at 40°C±5°C for at least 8 hours until the product concentration (i.e. ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopent[b]indol-3-yl)acetic acid) >40 area % by HPLC. Then, while stirring the mixture at 25°C±5°C, the pH value is adjusted to 4-5 by adding a 1.0 M aqueous solution of citric acid. Solids were removed by filtration, acetonitrile was removed by vacuum distillation, after which the product mixture was partitioned between ethyl acetate (40.8 kg), water (44.6 kg) and brine (6.7 kg). The ethyl acetate phase is washed with brine (2 x 54 kg) at least twice until the pH of the brine wash solution changes from 4 to 6 and the citric acid content of the ethyl acetate phase as determined by 1H NMR spectroscopy is be <0.1%. Ethyl acetate is replaced with 2-propanol (106 kg) using vacuum distillation. After complete removal of ethyl acetate, which was verified by 1H NMR spectroscopy, the 2-propanol solution of the product was diluted with 53.2 kg of 2-propanol, filtered and analyzed using achiral and

- 60 045975 chiral HPLC for determining the concentration of ©)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and the corresponding enantiomer of the acid (i.e. ©-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid) .

A mixture of the product in 2-propanol containing approximately equal amounts of ©)-2-(7-(4-cyclopentyl3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and the corresponding (S) ethyl ester, adjusted to a concentration of (©-acid 16.7 l/kg by adding 2propanol and stirred at 60°C±5°C. Seed material of L-arginine salt of compound 1 (0.18 kg) is added. An aqueous solution of L-arginine (2.27 M, 3.63 kg, 20.8 mol, 0.463 eq. in 9.12 kg of water, preheated to 60 ^° C ± 5 ° C) is added to the mixture to crystallize L-arginine salts of compound 1. The mixture is slowly cooled to 25°C±5°C, after crystallization is complete, the L-arginine salt of compound 1 is isolated by filtration and washed with 2-propanol (45 kg) and then ethyl acetate (97.4 kg), until until the impurity content of the wet filter cake reaches less than 1% by area according to HPLC.The product is then vacuumed at 40°C±5°C, sieved and packaged under nitrogen to obtain 12.15 kg (yield 42 .7%) unground L-arginine salt (R)-2-(7-(4-cyclopentyl-3 -(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)vinegar noic acid. Its morphology was found to be different from that of the product of Example 1. The morphology was characterized as a very fine powder consisting of small, densely layered agglomerates, fine flakes and very small irregular fragments (see Fig. 3A and Fig. 3B). HPLC: 99.5% by area (achiral); DSC: 204.11°C (melting point); particle size: 14 µm (average length), 1.94 µm minimum and 54.79 maximum, according to OPM.

Four lots (i.e. Lots J1, J2, J3 and J4) of L-arginine salt of Compound 1 totaling 45.82 kg were prepared from 88.4 kg of ethyl 2-(7-(4-cyclopentyl-3-(trifluoromethyl) )benzyloxy)-1,2,3,4-tetrahydrocyclopenta3-yl)acetate, wherein the preparation of Lot J2 is described directly above and Lots J1, J3 and J4 are prepared substantially in accordance with the procedure described for Lot J2. The yield of dry unground L-arginine salt from ethyl ester varies from 39.4% (mol) to 43.6% (mol) and averages 42.2% (mol). The total amount of impurities according to achiral HPLC ranges from 0.5 area% to 1.2 area%, with an average of 0.8 area%. The content of the corresponding ©-acid is less than the limit of quantitation (0.7% by area).

In addition to lot J2, the morphology of lot J1 is also analyzed using microscopy. The morphology of Lot J1 is characterized by thin lamellar flocs layered into agglomerates that are densely laminated and the agglomerates are not very porous (see Fig. 4A and Fig. 4B). There are differences in the morphology of lots J1 and J2.

All four lots were recrystallized from aqueous 2-propanol as described below in Example 3.

Example 3: Preparation of platelet crystal form or morphology by recrystallization of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole -3-yl)acetic acid

Suspension of L-arginine salt of ©-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[B]indol-3-yl)acetic acid (8.20 kg, 12.981 mol) in a mixture of 2-propanol (34.35 kg) and USP water (7.71 kg) is heated to 75°C with stirring at 100 rpm under nitrogen to produce a solution, which is then heated to 78°C and kept at the specified temperature for 0.5 hours. The solution is then cooled to 66°C over 1.5 hours and stirred at 61°C to 66°C for approximately 0.75 hours. 2-propanol (10.05 kg) was added over 1 hour 15 minutes and the mixture was stirred at 67°C for 1 hour. The suspension is heated to a temperature of 72°C to 74°C to obtain a solution with a few undissolved particles and the resulting solution is stirred at 81 rpm for 39 minutes at the specified temperature. The suspension was cooled to 67°C over 61 minutes, stirred at 67°C for 34 minutes and 2-propanol (10.06 kg) was added over 1 hour 11 minutes, maintaining the temperature at 65°C. The suspension is stirred at 66°C for 1 hour. The suspension is then heated to a temperature of 70°C to 74°C for 55 minutes to leave some undissolved crystals. The suspension is then cooled to 65°C over 1 hour 51 minutes and stirred at 60°C to 65°C for an additional 55 minutes, after which 2-propanol (10, 71 kg). The suspension is stirred at 66°C for 1 hour 57 minutes. The suspension is then reheated to 75°C for 1 hour, stirred at 75°C to 78°C for 40 minutes and gradually cooled to 67°C over about 1 hour. The suspension is stirred at a temperature of about 66°C to 67°C for 34 minutes, then cooled again to 20°C over 10 hours 48 minutes and stirring is continued for an additional 6 hours 10 minutes. The suspension is filtered. The filter cake is washed with 19.31 kg of 2-propanol. Filtration is easy. The wet residue is dried in vacuo at about 40° C. to obtain 7.44 kg (91% yield) of the L-arginine salt of Compound 1 as an off-white solid. The solid product is ground using a Fitz mill at 1000 rpm using pe

- 61 045975 hammer forward position. Ground L-arginine salt (7.36 kg, yield 89.8%) is packaged for storage. In fig. Figure 5A shows a microphotograph of the wafers obtained by optical polarization microscopy (OPM). DSC: 207.19°C; OPM: average length: 80 µm, (standard deviation 65 µm), minimum size 12 µm, maximum size 266 µm. In fig. 5B is a copy of the photomicrograph of FIG. 5A, wherein the outline of a substantially intact/complete free plate of an elongated hexagonal shape is added.

Another four lots of L-arginine salt (44.95 kg, prepared as described in Example 2) were recrystallized to give five lots of L-arginine salt with a total weight of 33.19 kg. Lot A3 is prepared as described immediately above. Lots A1, A2, A4 and A5 are produced substantially in accordance with the procedure described for Lot A3. For these lots, the total amount of impurities according to achiral HPLC is approximately 0.1% by area, and the achiral HPLC analysis shows 100.0% w/w. - 101.2% w/w on average 100.3% w/w. The content of the corresponding ^)-acid is less than the limit of quantification - 0.14% by area. The temperatures of the beginning of the phase transition of five lots, obtained during the study using DSC, are given below in the table. A1.

Table A1

Lot number of L-arginine salt Temperature of the beginning of phase transition in accordance with DSC Differential scanning calorimetry Dedicated morphology A1 ^a 208.09°C Figure 6 Plates A2 ^a 207.20°C Figure 7 Plates AZ ^b 207.19°C Figure 8 Plates A4 ^a 207.83°C Figure 9 Plates A5 ^a 207.90°C Figure 10 Plates

^a Obtained in accordance with the procedure described in example 3 ^b Obtained in example 3

Example 4: Preparation of crystalline platelet form or morphology of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3yl )acetic acid from (K^)-ethyl-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

This example describes three methods: method 1, method 2 and method 3. In each method, the free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1, 2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid is obtained by enzymatic hydrolysis of the corresponding (K^)-ethyl ester to K^)-acid, followed by modification of the procedure for obtaining L-arginine salt, previously described in WO2011/094008 . The isolation of the L-arginine salt directly as a free-lamellar crystalline form without a recrystallization step is then described.

These methods make it possible to obtain a free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3yl )acetic acid starting from the corresponding (K^)-ethyl ester without any direct isolation of the intermediate product.

Example 4.1. Method 1: Preparation of crystalline platelet morphology of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid

A solution containing 942 mg (6.92 mmol) of potassium phosphate monobasic dissolved in 7.0 g of water is added to a solution of potassium phosphate dibasic (12.0 g, 68.90 mmol) in 69.2 g of water. Testing the buffer solution shows a typical pH value of 7-8.

Add (I/8)-ethyl2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole- 3-yl)acetate (100.0 g, 206.0 mmol) and acetonitrile (700.0 g, 886 ml) to obtain a suspension. The flask is equipped with a reflux condenser and filled with nitrogen. A solution of potassium phosphate buffer is added to the suspension, followed by the addition of 4.0 g of lipase B (Candida antarctica, immobilized). Checking the pH value shows a typical value of 7-8. The reaction mixture is heated to a temperature of 40°C to 45°C until the content of ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid will not be >45 area % (but less than 50 area %), according to HPLC. The pH of the reaction mixture is adjusted to 4-5 by slowly adding a solution containing 9.0 g (46.84 mmol) of citric acid dissolved in 47.0 g of water. The solids are filtered off and washed with 150 g (190 ml) acetonitrile. The filtrates are combined and concentrated in vacuo to 101 g on a rotary evaporator (bath temperature 55°C, pressure 25 mmHg). The residue is dissolved in 200 g (222 ml) ethyl acetate. After this, 200 g of water and then 39.5 g of sodium chloride are added with vigorous stirring. After stirring for 5 minutes, phase once

- 62 045975 are separated and the organic layers are washed with sodium chloride solution (3x150 ml, each solution prepared using 100 ml saturated sodium chloride solution and 50 ml water). The organic phase is concentrated to 107 g in vacuo on a rotary evaporator (bath temperature 55°C, pressure 20 mmHg). The residue is dissolved in 80 g (101 ml) 2-propanol and concentrated to 140 g in vacuo on a rotary evaporator (bath temperature 55°C, 20 mmHg). The residue is diluted with 2-propanol to a mass of 300 g (add 160 g of 2-propanol, 202 ml). The solution is passed through a medium porosity filter and then transferred to a 1 liter jacketed reactor. The reactor is equipped with a top drive curved blade agitator, a reflux condenser, an internal temperature sensor and a MiniHuber thermostatic heating/cooling controller. When transferring, use an additional 100 g of 2propanol. The solution of the product, which contains 300 g (379.7 ml) of 2-propanol, is clear and contains no solids or cloudiness. 23.4 g of water were added to the solution with stirring, followed by the addition of solid L-arginine (17.94 g, 103.0 mmol, 0.5 eq. based on (R/S)-ethyl ester). The reaction mixture is heated to an internal temperature of 40°C. The consistency of the reaction mixture changes after 5 minutes, and then the mixture is heated to an internal temperature of 76°C. Water is added dropwise to the suspension until all solids are almost completely dissolved (31.5 d). The internal temperature is brought to 80°C to obtain a solution. While maintaining the internal temperature at 83°C, 3.3 g of L-arginine salt is added to the mixture for inoculation. L-arginine salt dissolves in solution. The resulting solution was cooled to an internal temperature of 70°C while stirring, and 2-propanol (40.3 g, over 20 minutes) was added dropwise to the solution until the solution became cloudy. The mixture is heated to an internal temperature of 75-80°C to obtain a clear solution. An additional 1.2 g of L-arginine salt is added to the solution for inoculation. 2-propanol (45 g, over 90 minutes) is added dropwise to the seed mixture. The resulting suspension is kept at 80°C for 10 minutes before adding 2-propanol again. Three additions are made (79.0 g, 78.5 g and 79.1 g), each over a period of approximately 75 minutes with a 10 minute break in between. After the last addition, the temperature controller ensures, in accordance with the specified program, cooling of the jacket from 85°C to 20°C within 6 hours and then maintaining the specified temperature.

The resulting mixture, while stirring, is subjected to two cycles of heating to an internal temperature of 70°C to 72°C and cooling to 20°C for 4 hours. After the second cycle, the mixture is kept at 20°C and the sample is examined using OPM. OPM shows the presence of plates only (see Fig. 11). The mixture is heated to an internal temperature of 55-60°C, maintained at this temperature for 1 hour and then slowly cooled (3 hours) to 30°C. After 30 minutes of exposure, the mixture is heated to 50-55°C, maintained at the specified temperature for 1 hour, then slowly cooled (3 hours) to a temperature of 20°C and maintained at the specified temperature.

Solids are collected by reduced pressure filtration (60 mm Hg below atmospheric pressure) using medium porosity filter paper. The filter cake is washed (re-suspended) with 125 g of 2-propanol, followed by a wash with 75 g of 2-propanol. The semi-dry filter cake is washed (re-suspended) with 100 g of ethyl acetate, followed by a wash with 100 g of ethyl acetate. The precipitate is dried in a vacuum oven (40°C, high vacuum) to obtain 65.2 g of L-arginine salt as an off-white solid (plates see Fig. 12A), the DSC curve of which shows an onset temperature of 205.6 °C (Fig. 12B). Yield = 100x((65.2 g - 4.5 g of seed) /65.05 g = 93.3% based on ^)-ethyl-2-(7-(4-cyclopentyl-3-(trifluoromethyl) benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (i.e., half the racemic mixture of the original ethyl 2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy) -1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetate). Chiral HPLC analysis shows that the content of compound 1 is 99.61% (by area).

Example 4.2. Method 2: Preparation of platelet crystal morphology of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid

A solution of 1.0 M buffer (pH 7-8) is prepared by mixing a solution of potassium phosphate monobasic (942 mg, 6.92 mmol) in 7.0 g water with a solution containing potassium phosphate dibasic (12.0 g, 68.90 mmol ) in 69.2 g of water.

In a 2-liter three-neck flask equipped with a magnetic stirrer and a reflux condenser, under a nitrogen atmosphere, ethyl 2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta3-yl)acetate (100.0 g, 206.0 mmol, 1.000 eq.) was suspended in acetonitrile (700.0 g). A 1.0 M solution of potassium phosphate buffer is added to the suspension, followed by the addition of 3.1 g of lipase B (Candida antarctica). Checking the pH shows a typical value of 7-8. The reaction mixture is then heated in an oil bath at 45°C until achiral HPLC analysis shows that the content of (R)-2-(7(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2. 3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid accounts for more than 44% (by area).

The reaction mixture is cooled to room temperature and the pH value of the reaction mixture is equal to

- 63 045975 up to 4-5 by slowly adding a solution containing citric acid (9.0 g, 46.84 mmol) dissolved in 47.9 g of water. The solids are collected by filtration and washed with acetonitrile (150 g). The filtrates are combined and concentrated in vacuo to 121.4 g on a rotary evaporator (bath temperature 55°C, pressure 141 mm Hg). The residue is dissolved in 200 g of ethyl acetate. 200 g of water are added to the mixture with vigorous stirring, followed by the addition of 39.5 g of saturated sodium chloride solution. After stirring for 5 minutes, the phases are separated and the organic layer is washed successively three times with solutions each containing 100 ml of saturated sodium chloride solution and 50 ml of water. The organic layers are concentrated in vacuo to 116.9 g on a rotary evaporator (bath temperature 55°C, pressure 139 mm Hg). The residue is dissolved in 180 g of 2-propanol. The mixture is concentrated. The residue is again treated with 180 g of 2-propanol and concentrated in vacuo on a rotary evaporator to 121.1 g (same conditions as for the ethyl acetate phase). The residue is diluted with 2-propanol (113 g) to a mass of 234 g. The solution is passed through a medium-porosity filter and then transferred to a 1-liter jacketed reactor. The reactor is equipped with a top drive curved blade agitator, a reflux condenser, an internal temperature sensor and a MiniHuber thermostatic heating/cooling controller. Additional 2-propanol (187.3 g) was used for washing and transfer. The total amount of 2-propanol is 300.1 g. The material is a solution.

L-arginine (17.94 g, 103.0 mmol, 0.500 eq based on (R/S) ethyl ester) followed by water (53.8 g) was added to the solution with stirring. The reaction mixture is heated to an internal temperature of 81°C for 1 hour 15 minutes to obtain a clear solution. At this stage, the water content of the aqueous 2-propanol mixture is 15.2% w/w. At a mass temperature of 81°C, 2.01 g of seed crystals (L-arginine salt of compound 1) are added. The seed crystals do not dissolve completely. The mixture is stirred for 15 minutes at 80-81°C. 2propanol (329.3 g, over 7 hours 30 minutes) was added to the seed crystal mixture while stirring at 150 rpm using a metering pump. At this stage, the water content in the aqueous 2-propanol mixture is 7.9% w/w. The stirring speed is increased to 180 rpm. After adding 2-propanol, the temperature controller ensures, in accordance with the specified program, that the jacket is cooled from 80°C to 23°C within 5 hours, followed by maintaining the temperature at 23°C.

After stirring overnight, a sample of the reaction suspension (500 mg) is centrifuged. The solid is suspended in 0.5 ml of ethyl acetate, centrifuged and the resulting solid is dried at 40°C in a vacuum oven at 100 mmHg. Overlaying the PXRD XRD patterns of the sample with the PXRD XRD patterns of Lot A5 (plates, see Table A) shows that the sample represents the same crystalline phase as Lot A5 (FIG. 13). OPM micrographs show a predominantly lamellar morphology with lamellar sizes up to ~200 μm, a few lamellar aggregates, and lamellar fragments (Fig. 14).

The reaction suspension is subjected to two cycles of heating to an internal temperature of 75-78°C, maintaining at this temperature for 45 minutes and cooling to 24°C for 5 hours. After the first cycle, a 500 mg sample of the mixture is maintained at 24°C, centrifuged and the resulting solids are suspended in 0.5 ml of 2-propanol. The solids obtained by centrifugation from 2-propanol were resuspended in 0.5 ml ethyl acetate, centrifuged, dried at 40°C in a vacuum oven at 100 mmHg and their morphology was examined. OPM micrographs show a predominantly lamellar morphology with few aggregates. DSC shows bimodal melting. The mixture in the jacketed reactor is maintained at 24°C with a stirring speed of 180 rpm.

Solids are isolated by filtration at slightly reduced pressure using medium porosity filter paper. The initial fluid removal time is 90 minutes. The filter cake is washed with 79 g of 2-propanol (resuspension method) and 79 g of 2-propanol (displacement method). The total removal time for 2-propanol washes is 45 minutes. The semi-dry precipitate is washed with 90.2 g of ethyl acetate (resuspension method) and 90.2 g of ethyl acetate (displacement method). The total time to remove the ethyl acetate wash liquid is 40 minutes. The precipitate was dried in a vacuum oven (at 40°C and 120 mm Hg) to obtain 60.4 g (yield 46.45%, based on (R/S)-ethyl ether used for enzymatic separation, and with adjusted for the added amount of L-arginine salt seed crystals of compound 1) L-arginine salt of compound 1 as an off-white solid. Chiral HPLC analysis shows that the content of compound 1 is 99.78% (by area).

Example 4.3. Method 3: Preparation of platelet crystal morphology of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid (lot 06GSp)

Mixture of (K©-ethyl-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta3-yl)acetate (100.0 g, 206.0 mmol, 1.000 eq.), lipase B (4.0 g, Candida antarctica, immobilized) and an aqueous solution of potassium phosphate buffer (pH 7.8 ± 0.2, pH = 8.0 for this load) in acetonitrile

- 64 045975 (886 ml) is stirred under a nitrogen atmosphere at 40°C±5°C for at least 6 hours (24 hours for this load) and until the content(s)-2-(7-(4-cyclopentyl) -.3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid > 40% by area, HPLC; 48.4 area% for this loading. Then, with stirring at 25°C±5°C, the pH value is adjusted to 4-5 by adding an aqueous solution of citric acid (9.0 g). Solids were removed by filtration, acetonitrile was removed by distillation in vacuo and the resulting product mixture was partitioned between ethyl acetate (222 ml) and water (200 ml). The ethyl acetate phase is washed with brine at least twice until the pH value reaches 4 or more. Ethyl acetate is replaced with isopropanol (twice, 101 ml and 2 02 ml) using vacuum distillation. After complete removal of ethyl acetate (GC control, <0.5% by area, 0.41% for this load), the product solution in isopropanol is heated to 60°C±5°C, then cooled to <40°C, filtered and analyzed with using HPLC to determine the concentration of ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]in dol-3-yl)acetic acid (free carboxylic acid acid). Isopropanol is added and removed (vacuum distillation to obtain a solution of N-free carboxylic acid in isopropanol with a concentration in the range of 11-14 wt.%; for this load, 12.4%).

The isopropanol solution of the product is treated with the calculated amount of water (23 g) and L-arginine (15.9 g) and heated to 83°C±2°C. At this temperature, add water (28.1 g) until all solids have dissolved. The solution is then cooled to 73°C ± 2°C and treated with isopropanol (26.1 g) to obtain a cloudy solution. The solution is heated to 75°C±2°C and seed crystals of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydro are added to the solution cyclopenta[b]indol-3-yl)acetic acid (1.2 g). Isopropanol (280 g) is added to the seed mixture over at least 1 hour. The reaction mixture is then cooled to 20°C±2°C for 6 hours.

Heating/cooling cycles are carried out as follows. The product mixture is heated with stirring to 71°C±2°C and then cooled over 4 hours to 21°C±2°C. The mixture is then heated to 60°C±2°C, stirred at this temperature for 60±10 minutes, and then cooled to 31°C±2°C over 3 hours. After stirring at 31°C±2°C for 25±5 minutes, the stirred mixture is heated to 50°C±2°C, stirred at this temperature for 45±5 minutes, and then cooled to 20° over 3 hours C±2°C. Heating/cooling cycles can be represented as 71°C to 21°C to 60°C to 31°C to 50°C; temperature ±2°C compared to the cycles of other lots presented in table. 2A. Stirring at 20°C±2°C is continued for at least 2 hours until the suspension is filtered.

Solids are recovered by filtration (filtration works well), the filter cake is washed first by resuspension with isopropanol (100 g), then displacing with isopropanol (100 g), then resuspending with ethyl acetate (100 g), and finally displacing with ethyl acetate (100 g). G). HPLC analysis does not show impurities > 1% by area. The product is dried in vacuum at 40°C±5°C to constant weight (to loss on drying (LOD) <2% for 12 hours) to obtain 54.9 g L-arginine salt ^)-2-(7- (4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4tetrahydrocyclopenta[b]indol-3-yl)acetic acid as off-white crystalline plates (100.0% by area, HPLC, L content -arginine 27.5%, chiral purity 99.67%).

Three additional lots (portions) are obtained in accordance with the method described above, but with some differences, which are presented in table. A2.

- 65 045975

Table A2

Lot 04GSp Lot 05GSp Lot 07GSp Conditions for lipase hydrolysis 20 hours, 40°C ± 5°C pH=8.0; (R)-acid=47.1% by area 22 hours, 40°С ± 5°C; pH=8.0; (R)-acid=48.4% by area 21 hours, 40°С ± 5°С; pH=8.0; (R)- acid=47.9% by area Concentration: (R)-acid in FAI 12.4% w/w 12.4% w/w 11.7% wt./mass. Heating/cooling cycles A A IN Product filtration good good good Ultimate L-apr. salt (R)-acids 99.8% by area (purity) Yield: 54.9 g Off white crystalline plates Contents L-apr.: 30.1% Chiral purity: 99.63% 99.8% by area (purity) Yield: 56.9 g Off white crystal plates Contents L-apr: 27.8% Chiral purity: 99.52% 100.0% by area (purity) Yield: 54.7 g Off-white crystalline plates Contents L-apr: 27.4% Chiral purity: 99.62% А=70°С to 20°С to 70°С to 20°С to 60°С to 31°С to 50°С; ± 2°С В=70°С to 20°С to 70°С to 20°С to 70°С to 20°С to 60°С to 31°С to 50°С; ± 2°С

The temperatures of the onset of the DSC phase transition for all four lots are presented below in the table. A3. ______________________________Table A3______________________________

Lot No. Lot number Larg. salt Temperature of the beginning of phase transition in accordance with DSC Differential scanning calorimetry curve Dedicated morphology 04GSp ^b Ab 206.55°C Figure 22 Plates 05GSp ^b A7 206.94°C Figure 2 3 Plates 06GSp ^a A8 207.04°C Figure 2 4 Plates 07GSp ^b A9 207.63°C Figure 2 5 Plates

^a Obtained in Example 4.3 ^b Obtained essentially in accordance with the procedure for obtaining 0 6GSp and table. A2

Example 5: Preparations - assessment of the stability of spherulite and plate preparations

In order to evaluate whether two morphologies, spherulites and platelets, have an effect on the stability of the drug in the form of tablets of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2, 3 ,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid, two batches of tablet cores are prepared using the same composition, which is described in table. B, and using the direct compression process described in FIG. 15.

- 66 045975

Table B

Component Function wt./mass. mg/tab. Weight (g) L-apr. salt connection 1 FAI 1.40 1.40 7.00 Mannitol 200 SD, USP Diluent 54.10 54.10 270.50 Avicel РН102, NF Diluent 40.00 40.00 200.00 Explotab, N.F. Disintegrating substance 4.00 4.00 20.00 Magnesium stearate, NF Lubricating substance 0.50 0.50 2.50 Total 100.00 100.00 500.00

Batch weight (g)=500; Tablet weight (mg)=100

In table C and D show the results of the stability assessment under accelerated aging conditions. After 6 months of storage under accelerated aging conditions, the total amount of impurities (ie the total percentage of the two impurities represented by Compound A and Compound B) was 0.93% for the spherulite preparation and 0.51% for the plate preparation. The chemical stability of L-arginine salt with plantate morphology in tablet form is superior to the chemical stability of the preparation of L-arginine salt with spherulitic morphology in tablet form.

Results of a study of the stability of the spherulitic morphology of L-arginine salt (R)-2-(7-(4cyclopentyl-3 -(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid after 6 months storage at 40°C/75% RH are shown in table. C.

_______________________________Table C______________________________

Stability of spherulitic morphology

Index Original value 40°C/75%RH 1 month 3 months 6 months Content, % 96, 9 97.9 96, 8 94.5 Compound A (%) 0.06 0.17 0.68 0.61 Compound B (%) 0.04 0.06 0.19 0.32 Total impurities 0.10 0.23 0.87 0.93

Results of a study of the stability of the lamellar morphology of L-arginine salt (R)-2-(7-(4cyclopentyl-3 -(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid after 6 months storage at 40°C/75% RH are shown in table. D.

_______________________________Table D_______________________________

Stability of lamellar morphology

Index Original meaning 40°C/75%RH 1 month 3 months 6 months Content, % 99, 9 98.8 101.5 97.3 Compound A (%) 0.06 0.10 0.60 0.31 Compound B (%) 0.05 0.06 0.17 0.20 Total impurities 0, 11 0.16 0.77 0.51

Two batches of tablet cores (1 mg and 5 mg, using L-arginine salt lamellar morphology) are prepared in accordance with the composition described in table. B (above), using the same direct pressing method as shown in FIG. 15. Both drugs are packaged in 40 cc HDPE bottles. cm with dehumidifier. In table E and F present the results of a long-term stability study of the resulting tablets over 24 months at 25°C and 60% RH.

- 67 045975

Table E

Stability of tablets (lamellar morphology) (1 mg FAI) after 24 months Index Original value 25 °C/60% RH 6 months 9 months 12 months 18 months 24 months Content, % 97.5 96, 8 97.2 94.5 96.5 97.2 Compound A (%) 0.11 0.18 0.20 0.21 0.22 0.25 Compound B (%) <QL 0.05 0.05 <QL 0.06 <QL Total impurities ³ 0.11 0.23 0.25 0.21 0.28 0.25

^a Total percentage of compound A and compound B QL - limit of quantitation (0.05%)

Table F

Stability of tablets (lamellar morphology) (5 mg FAI) after 24 months Index Original value 25 °C/60% RH 6 months 9 months 12 months 18 months 24 months Content, % 99.8 98, 8 100.2 96, 8 97.5 99.3 Compound A (%) 0.06 0, 12 0.13 0.16 0.17 0.19 Compound B (%) <QL ND ND <QL <QL <QL Total impurities ³ 0.06 0, 12 0.13 0.16 0.17 0.19

^a Total percentage of compound A and compound B

QL - limit of quantitation (0.05%); ND=He detected

Example 5B: Preparations of L-arginine salt of ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid

Tablet cores are prepared in accordance with the composition described in table. G, and in accordance with the method substantially illustrated in FIG. 15. The amounts of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid were calculated taking into account the content of compound 1 in the original salt is 72.42%.

Table G

Dose per tablet 0.5 mg 1 mg 2 mg 3 mg L-apr. salt of compound 1 0.69 1.381 2,762 4, 143 Mannitol Pearlitol® 100SD 54.81 54.119 52,738 51,357 Avicel® Microcrystalline Cellulose 40 40 40 40 Sodium starch glycolate Explotab® 4 4 4 4 Magnesium stearate 0.5 0.5 0.5 0.5 Opadry® II Blue 4 4 4 4 Total tablet weight 104 104 104 104

Example 6: Comparison of the hygroscopicity of plates and spherulites using the method of dynamic moisture sorption (DMS)

Dynamic moisture sorption (DMS) analysis is carried out on a sample of lot A2 (see example 3, table A) of L-arginine salt of compound 1 lamellar morphology and on a sample of lot H2 (obtained in accordance with the procedure described in example 8: method 2, stage G and H, in WO2011/094008, as described in Table K, Example 8 of the present invention) L-arginine salt of Compound 1 with spherulite morphology. DSW analysis carried out at 25°C shows that the L-arginine salt of compound 1 (lamellar morphology) is not hygroscopic and the weight increase at RH 90% is only 0.2 wt.% (Fig. 16 and Fig. 17). However, the increase in mass of L-arginine salt of compound 1 (spherulitic morphology) is 0.5% by mass (adsorption cycle) and 0.7% by mass (desorption cycle) at 90% RH and 25°C. The results show that the lamellar morphology has improved hygroscopicity compared to the spherulitic morphology.

Example 7: Comparison of Powder X-Ray Diffraction of Wafers and Spherulites

Powder X-ray diffraction (PXRD) shows that Lots A1 A5 of Example 3 (see Table A) are crystalline with similar X-ray patterns, indicating that they are of the same crystalline phase (see FIG. 18). PXRD analysis is carried out on samples of lot A2 (see example 3, table A) L-arginine salt of compound 1 lamellar morphology and a sample of lot H2 (obtained by the method described in example 8, method 2, steps G and H in WO2011/094008, and in Table K

- 68 045975 (example 8) in the present invention) L-arginine salt of compound 1 of spherulite morphology. A comparison of Lot A2 (lamellar morphology) and Lot H2 (spherulitic morphology) shows that both lots represent the same crystalline phase, however Lot A2 shows better diffraction and lower background scattering (i.e. less amorphous halogen contribution), which indicates a higher degree of crystallinity of the plates compared to spherulites (Fig. 19).

Some peaks of the powder X-ray diffraction pattern of Lot A2 L-arginine salt of compound 1 (plates) are presented in Table. H below.

_________________________________Table H_________________________________

Field [°2θ] Height [units] Interpl. distance [A] Rel. INT. [%] 8.2 33633.8 10.8159 93, 6 14.7 402.2 6.0252 1.1 15.6 364.5 5.6724 1.0 16, 4 4734.0 5.4187 13.2 16, 6 931.2 5.3288 2.6 17.2 390.6 5.1667 1.1 19.1 927.9 4.6471 2.6 20.1 994.1 4.4250 2.8 20.5 35918.7 4.3373 100.0 22.0 1056.0 4.0463 2.9

Field [°2θ] Height [units] Interpl. distance [A] Rel. INT. [%] 22.2 645.6 4.0046 1.8 22.9 811.2 3.8855 2.3 23.2 671.6 3.8404 1.9 23.4 1033.2 3.8038 2.9 23.9 1130.9 3.7195 3.2 24.6 3586.1 3.6159 100 25.9 356.9 3.4455 1.0 26, 4 363.3 3.3785 1.0 28.8 1899.7 3.1004 5.3 37.3 1922.3 2.4124 5.4

Some X-ray powder diffraction peaks of Lot H2 L-arginine salt of compound 1 (spherulites) are presented in Table. I below.

_____________________________________Table I_____________________________________________

Field [°2θ] Height [units] Interpl. distance [A] Rel. INT. [%] 8.2 4469.9 10.8266 94, 9 11.9 1777.7 7.4137 37.7 12.5 1409.5 7.0965 29, 9 12.9 1340.9 6.8507 28.5 13.1 1152.3 6.7346 24.5 13.9 868.3 6.3787 18.4 14.7 844.0 6.0348 17, 9 15.6 750.7 5.6626 15, 9 16, 6 1356.8 5.3260 28.8 17.1 349.1 5.1758 7.4 19.0 1954.6 4.6610 41.5

Example 8: Evaluating methods of obtaining

Field [°2θ] Height [units] Interpl. Distance [A] Rel. INT. [%] 20.0 1752.3 4.4293 37.2 20.5 4712.7 4.3383 100.0 20.9 2525.3 4.2584 53.6 22.0 1799.9 4.0432 38.2 22.6 1326.4 3.9363 28.2 23.2 1686.9 3.8367 35.8 24.0 912.9 3.7137 19.4 24.6 953.2 3.6193 20.2 25.2 386.8 3.5281 8.2 28.9 90.5 3.0857 19 30.6 167.1 2.9198 3, 6

L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid

Some methods for the preparation of L-arginine salt of L-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[L]indol-3-yl)acetic acid have been described earlier [see WO2010/011316 (example 1.33) and WO2011/094008 (example 7, method 1, stage B; example 8, method 1 and method 2 stage H).

OPM shows that the morphology of the L-arginine salt of compound 1, obtained in accordance with the method described in example 1.33 (WO2010/011316), contains aggregates and spherulites with an admixture of small crystalline particles that are too small to determine their morphology. Overall, the sample is weakly crystalline and no plates are identified. The same procedure is also described in WO2011/094008 (see example 8, method 1). In this procedure, L-arginine salt is obtained starting from practically pure L-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3- yl)acetic acid.

OPM shows that the morphology of the L-arginine salt of Compound 1 obtained in Example 7 (Method 1, Step B) presented in WO2011/094008 includes spherulites with some fine particles (see FIG. 20). The particle size of the sample, determined using OPM, is presented in Table. J. In this example, the L-arginine salt of compound 1 is prepared in the presence of L-ethyl ester, which remains after the enzymatic hydrolysis step.

__________________________________________Table J__________________________________________

Sample Average value Standard deviation Minimum Maximum Example 7, method 1 stage B ¹ 18, 05 12.18 5.43 58.37

¹ WO2011/094008

Similarly, as described in example 7 (method 1 step B), the L-arginine salt of compound 1 is prepared from (K^)-ethyl-2-(7-(4-cyclopentyl-3-(trifluoromethyl)- benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate, as described in WO2011/094008 (example 8, method 2 steps G and H). Six lots of L-arginine salt of compound 1 are obtained, of which the preparation of one lot is specifically described in Example 8 (method 2, steps G and H, i.e., preparation of lot H1), and five additional lots (i.e., lots H2 - H6) L-arginine salt of compound 1 is prepared using essentially the same procedure. The lots are shown to represent the same crystalline phase spherulitic morphology with an onset melting temperature ranging from 203.00°C to 203.97°C as determined by differential scanning calorimetry (DSC) (see Table K).

Table K

Lot number Morphology Method of obtaining Pace. started Dedicated morphology H1 Spherulite W02011/094008 ¹ 203.46°C Spherulite H2 Spherulite W02011/094008 ² 203.9b°C Spherulite NZ Spherulite W02011/094008 ² 203.00°C Spherulite H4 Spherulite W02011/094008 ² 203, 1GS Spherulite H5 Spherulite W02011/094008 ² 203.79°C Spherulite H6 Spherulite W02011/094008 ² 203.97°C Spherulite

¹ see Example 8, method 2 stages G and H 2

Received in accordance with the methodology for obtaining lot H1

Based on the six lots described in WO2011/094008, four (4) new additional lots (ie J1 - J4) are obtained according to essentially the same methodology (see Example 2). Each of these new lots is prepared from at least 21.9 kg of (R/S)-ethyl-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent[ b]indol-3-yl)acetate. Each lot is prepared using the selective enzymatic hydrolysis of (R/S)-ethyl-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetate to give the corresponding (K)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol3-yl)acetic acid (i.e. connections 1). In the next step, without direct isolation of compound 1, a preheated aqueous solution of L-arginine is added to the solution (i.e., the solution containing compound 1, ^-ethyl ether, isopropyl alcohol and water) to obtain the L-arginine salt of the compound 1. The crude mixture containing the L-arginine salt of Compound 1 is then subjected to a series of washings and filtrations as shown in FIG. 21. Two of the four lots (ie Lots J1 and J2) are analyzed by microscopy. The morphologies of these two lots are shown to be different (see example 2), and their morphologies are also shown to be different from the morphology or habit of the spherulites of the six lots described in WO2011/094008.

In addition to the difference in morphologies, it is also shown that each method used to prepare four lots of L-arginine salts of compound 1 from (R/S)-ethyl-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1 ,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate, leads to a number of undesirable deviations from the form of the six lots described in WO2011/094008. Once the L-arginine salt of Compound 1 is formed, the resulting product slurry is transferred to a filter dryer to begin a series of washing and filtration steps as shown in FIG. 21. Surprisingly, the filtration step is found to be excessively long, ineffective and ultimately leads to filter clogging. Filtration inefficiency results in the transfer of excess wet material from the filter dryer to the dryer trays, thus resulting in additional process delays due to long drying periods to achieve a given performance (i.e., drying weight loss <2% by weight). The approximate time required for filtration is presented in table. L.

Table L

Lot number Batch weight (kg) See Figure 21 Total filtration time (min.) ¹ Total filtration time 1 (min.) Total filtration time 2 (min.) Total filtration time 3 (min.) Total filtration time 4 (min.) l 22.16 31 82 935 378 1426 J2 ² 21, 86 80 228 1371 NR ³ 1679 J3 21, 80 43 703 6125 564 7435 J4 22.58 43 570 4643 625 5881

¹ Total filtration time 1-4 ² Preparation is described in example 2; Lots J1, J3 and J4 are obtained in the same way.

³ Not required

Surprisingly, it has been found that the new methods described in the present invention provide

- 70 045975 obtaining the L-arginine salt of compound 1 in a free-lamellar form instead of the morphology observed for the previous lots (i.e. the six (6) lots described in WO2011/094008, see table K; and the two studied lots, as described above (Lots J1 and J2), see Table L). Lamellar morphology offers a number of advantages such as easier filtration, higher degree of crystallinity, improved hygroscopicity and better drug stability.

Example 9: Powder X-ray diffraction

Powder X-ray diffraction (XRD) data are acquired on an X'Pert PRO MPD powder diffractometer (PANalytical, Inc.) with a Cu source set at 45 kV and 40 mA, Cu(Ka) radiation and an X'Celerator detector. The samples are placed in the sample holder and spread with a flat spatula and weighing paper. By rotating the samples, X-ray diffraction patterns are obtained with a 12-minute scan in the 2-theta range of 5-40°2θ. Radiographs were viewed and analyzed using X'Pert Data Viewer Software, version 1.0a and X'Pert HighScore Software, version 1.0b.

Example 10: Differential scanning calorimetry

Differential scanning calorimetry (DSC) studies were performed using a TA Instruments, Q2000 at a heating rate of 10°C/min. The instrument is calibrated for temperature and energy using the melting point and enthalpy of fusion of indium as a standard. Thermal phenomena (desolvation, melting, etc.) are assessed using Universal Analysis 2000 software, version 4.1D, Build 4.1.0.16. Samples are weighed in aluminum trays and scanned from ~25°C to ~270°C at a rate of 10°C/min.

Example 11: Thermal gravimetric analysis

Thermogravimetric analysis (TGA) was performed using a Q500 TA Instruments TGA or Q5000 at a heating rate of 10°C/min. The instrument is calibrated using a standard balance and alumel and nickel furnace standards (Curie point determination). Thermal phenomena such as mass loss are calculated using Universal Analysis 2000 software, version 4.1D, Build 4.1.0.16.

Example 12: Dynamic Moisture Sorption Analysis

DSW analysis was performed using a TA Instruments Q5000 SA (EQ2418). The characteristics of the device have been tested by the manufacturer. Samples are placed in a calibrated sample holder on a Q5000 SA balance. The sample is dried at 40°C and then analyzed at 25°C with an adsorption phase from 30% RH to 90% RH and a desorption phase from 90% RH to 10% RH.

Example 13: Microscopy

Microscopy studies are carried out using a Nikon Eclipse E600 POL (EQ0124) with a Nikon DS Fi1 (EQ0123) digital camera. Nikon software (NIS-Elements BR 3.0) is used to obtain microphotographs. The characteristics of the equipment are guaranteed by the manufacturer. Samples are prepared for analysis by placing a small amount of solid on a glass slide, adding a drop of mineral oil to the cover slip, placing the cover slip with the oil on it over the sample and pressing it gently with a gloved finger.

Example 14: Determination of specific surface area using the BET (Brunauer, Emmett and Teller) method

In general, the specific surface areas of lots A1-A9 (plates) and H1-H6 (spherulites) are determined by physical adsorption of nitrogen gas on the surface of a sample of each lot according to a well-known technique based on the theory of Brunauer, Emmett and Teller.

The BET specific surface areas of samples are quantified by Micromeritics Pharmaceutical Servies using a Micromeritics Surface Area Analyzer??? TriStar II BET (MicroActive for TriStar II Plus 2.02 Software™). Samples are degassed at 25°C for 960 minutes under vacuum (eg 100 mm/Hg). Quantitative determination of N2 adsorption at 77.3 K (-195.85°C) is carried out using the BET method of determining the surface area of eleven points with relative pressures ranging from about 0.05 to about 0.3 (P/Po) for a weighed amount each sample, see table. M1, tab. M2 and tab. N below. The analysis is carried out according to ISO9277.

Table M1

Lamellar morphology Arena lot Lot number Sample(g) Correlation coefficient BET surface area ( ^m2 /g) 5015-12-12 A1 0.6163 0.99916 0.7 5015-12-13 A2 1.5270 0.99945 0.7 5015-12-14 AZ 0.4465 0.99922 15 5015-12-15 A4 0.5709 0.99939 1, o 5015-12-16 A5 0.9582 0.99940 0.8

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Claims (14)

Table M2 Lamellar morphology Arena lot Lot number Sample (g) Correlation coefficient Surface area according to the BET method (m2/g) 04GSp Ab 0.4332 0.99921 2.4 05GSp A7 0.3652 0.99910 1.9 06GSp A8 0, 6866 0.99984 3, 0 07GSp А9 0,2754 0,99914 3, 1 04GSp Ab 0,4332 0,99921 2,4 Table N Spherulite morphology Arena lot Lot number Sample (g) Correlation coefficient Surface area according to the BET method (m2 /g) 5015-10-01 N1 0.8891 0.99967 10, 0 5015-10-02 N2 0.6766 0.99967 13.5 5015-10-03 NZ 0.7002 0.99966 11, 8 5015- 10-04 N4 0.9420 0.99964 12.0 5015-10-05 N5 0.7658 0.99955 10.9 5015-10-06 N6 0.9662 0.99965 12.3 It will be clear to specialists in this field of technology that various modifications, additions, substitutions and variations to the illustrative examples set forth herein may be made without departing from the spirit of the invention and thus within the scope of the present invention. CLAIM 1. Free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid, having an X-ray powder diffraction pattern having peaks, in units of 2θ, at: 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2°, 24, 6°±0.2°, 28.8°±0.2° and 37.3°±0.2°, where the free-plate crystal form is characterized by thin hexagonal plates, where two opposite sides of the plate are longer than the other sides; wherein the free-plate crystal form includes whole plates, substantially whole plates, plate fragments and mixtures thereof that are free of radial clusters or spherulites; where the radial cluster and spherulites are a crystal habit consisting of thin plates or flakes that are clustered radially around the nucleation site of the site, which show a cross when viewed with cross-polarized light. 2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.5°C to 208.1°C when scanning at a speed of 10°C/min; 2) on the differential scanning calorimetry curve there is an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.5°C to 208.5°C when scanning - 72 045975 rotation at a speed of 10°C/min; 2. The free-plate crystalline form of claim 1, having a BET specific surface area of from about 0.1 m ² /g to about 5.0 m ² /g; BET specific surface area from about 0.6 m ² /g to about 4.0 m ² /g; BET specific surface area from about 0.6 m ² /g to about 1.6 m ² /g. 3) a plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.2% by mass or less at 90% RH; and/or 3) a plot of dynamic moisture sorption (DMS) with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form of approximately 0.3% by mass or less at 90% RH; and/or 3. Free-plate crystal form according to any one of claims 1, 2, having a differential scanning calorimetry curve: containing an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.0°C to 208.5°C when scanning at a speed of 10°C/min; containing an endotherm with an extrapolated temperature of the beginning of the endothermic transition in the region from 205.5°C to 208.1°C when scanning at a speed of 10°C/min; or obtained by scanning at a speed of 10°C/min, containing an endotherm as substantially represented in any of FIGS. 6-10 and 22-25. 4) BET specific surface area ranges from about 0.6 m ² /g to about 4.0 m ² /g. 4) the BET specific surface area is from about 0.6 m ² /g to about 4.0 m ² /g; or B) 1) the X-ray powder diffraction pattern has peaks, in units of 2θ, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2°, 24.6° ±0.2°, 28.8°±0.2°, and 37.3°±0.2°; 4. The free-lamellar crystalline form according to any one of claims 1-3, wherein the dynamic moisture sorption (DMS) graph of which with an adsorption phase from 30% RH to 90% RH shows an increase in the mass of the free-lamellar crystalline form by approximately 0.3 wt. .% or less at 90% RH. 5. Free-plate crystal form according to claim 1, where A) 1) The powder X-ray diffraction pattern has peaks, in 2θ units, at 8.2°±0.2°, 16.4°±0.2°, 20.5°±0.2° and 24.6° ±0.2°; 6. A composition containing a free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3 -(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol- 3-yl)acetic acid according to any one of claims 1-5. 7. Pharmaceutical composition containing a free-plate crystalline form of L-arginine salt (R)-2-(7-(4-cyclopentyl-3 -(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta [b]indole-3 -yl)acetic acid according to any one of claims 1-5; and pharmaceutical excipient; optionally wherein the pharmaceutical composition is suitable for oral administration. 8. Use of the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid according to any one of claims 1 to 5 in a method of treating a person or animal with therapy. 9. Use of the free-plate crystalline form of L-arginine salt (R)-2-(7-(4cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl) acetic acid according to any one of claims 1-5 in the method of treatment: S1P ₁ receptor-related disorder, where optionally the S1P1 receptor-related disorder is selected from the group consisting of: lymphocyte-mediated disease or disorder, autoimmune disease or disorder, inflammatory disease or disorder, primary biliary cirrhosis, cancer, psoriasis, psoriatic arthritis , rheumatoid arthritis, Crohn's disease, transplant rejection, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, ulcerative colitis, type I diabetes, and acne; or where optionally the S1P1 receptor-related disorder is: a lymphocyte-mediated disease or disorder, or an autoimmune disease or disorder; or an inflammatory disease or disorder. 10. Method for obtaining a free-plate crystalline form of L-arginine salt (R)-2(7-(4-cyclopentyl-3 -(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3 - yl)acetic acid according to any one of claims 1-5, including the following stages: a) obtaining the first mixture containing L-arginine salt A)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid, water-miscible antisolvent and H ₂ O; b) heating the first mixture to the first heating temperature to obtain a second mixture; wherein said first heating temperature is from about 79°C to about 85°C; c) adding a first additional amount of said water-miscible anti-solvent to said second mixture while maintaining said first heating temperature to obtain a suspension; d) cooling the suspension to the temperature of the first cooling, followed by heating to temp. - 73 045975 second heating schedule; wherein said first cooling temperature is from about 18°C to about 22°C, and said second heating temperature is from about 69°C to about 73°C; e) cyclically repeating step d) optionally one or more times, where the first cooling temperature of each cycle may be the same or different, and the second heating temperature of each cycle may be the same or different; And f) cooling said suspension to a final cooling temperature to obtain said free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid; wherein said final cooling temperature is from about 18°C to about 22°C. 11. Method for obtaining free-plate crystalline form of L-arginine salt (R)-2(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3- yl)acetic acid according to claim 10, including the following steps: a) hydrolysis of the compound of formula (IIa): where R ³ represents ^-^-alkyl; in the presence of a hydrolysis mixture containing lipase and a solvent for the hydrolysis step to obtain da-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid; b) obtaining a first mixture containing L-arginine salt ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl )acetic acid, water-miscible antisolvent and H ₂ O; c) heating the first mixture to the first heating temperature to obtain a second mixture; wherein said first heating temperature is from about 79°C to about 85°C; d) adding a first additional amount of said water-miscible anti-solvent to said second mixture to form a slurry while maintaining the first heating temperature; And e) cooling said suspension to a first cooling temperature and then heating it to a second heating temperature; wherein said first cooling temperature is from about 18°C to about 22°C; and said second heating temperature is from about 69°C to about 73°C; f) cyclically repeating step e) optionally one or more times, wherein said first cooling temperature in each cycle may be the same or different and said second heating temperature in each cycle may be the same or different; And g) cooling said suspension to a final cooling temperature to obtain said free-plate crystalline form of L-arginine salt ^)-2-(7-(4-cyclopentyl-3(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent [b]indol-3-yl)acetic acid; wherein said final cooling temperature is from about 18°C to about 22°C. 12. Method according to claim 11, where i) said compound of formula (IIa) is said lipase is immobilized Candida Antarctica lipase B; and said hydrolysis step solvent contains acetonitrile; and/or ii) hydrolysis is carried out in the presence of a phosphate buffer at a pH of from about 6.9 to about 8.1, where the specified phosphate buffer is a potassium phosphate buffer; and/or iii) hydrolysis is carried out at a temperature of from about 35°C to about 45°C; and/or iv) after said hydrolysis, said ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid is not allocate; and/or optionally where, after said hydrolysis, said da-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid present in an amount of at least 40% as determined by HPLC; and/or v) wherein the water-miscible antisolvent includes a solvent selected from the group, - 74 045975 consisting of acetonitrile, acetone, tetrahydrofuran and C2-C ₄ alkanol; and/or optionally wherein the water-miscible antisolvent comprises 2-propanol; and/or vi) wherein said second mixture is a substantially homogeneous solution, and/or vii) the weight ratio of the L-arginine salt is ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1, 2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and said first additional amount of 2-propanol is from about 1.00:5.95 to about 1.00:6.25; and/or viii) wherein cooling of said suspension to said first cooling temperature in step e) is carried out at a rate of from about 9°C/hr to about 11°C/hr; and/or ix) wherein the cycling of step f) includes said cycling of step e) two times; optionally where step e) is repeated cyclically twice: cooling the suspension to a first cycle cooling temperature such as 16°C to 26°C; heating the slurry to a first cycle heating temperature such as 55°C to 65°C; cooling the suspension to a second cycle cooling temperature such as 26°C to 36°C; and heating the slurry to a second cycle heating temperature such as 45°C to 55°C; or wherein said cyclic repetition of step f) includes said cyclic repetition of step e) three times; optionally wherein cyclically repeating step e) three times includes: cooling the suspension to a first cycle cooling temperature such as 16°C to 26°C; heating the slurry to a first cycle heating temperature such as 66°C to 76°C; cooling the suspension to a second cycle cooling temperature such as 16°C to 26°C; heating the slurry to a second cycle heating temperature such as 55°C to 65°C; cooling the suspension to a third cycle cooling temperature such as 26°C to 36°C; and heating the slurry to a third cycle heating temperature such as 45°C to 55°C. 13. The method according to claim 11 or 12, wherein the preparation of said first mixture in step b) includes the step of adding L-arginine and H2O, together or separately in any order, to a salt-forming mixture containing ^)-2-(7-( 4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]yne dol-3-yl)acetic acid and said water-miscible antisolvent to obtain said first mixture; optional where: i) said addition of L-arginine and H2O is carried out under an inert atmosphere containing nitrogen; ii) before adding said L-arginine and H2O, said salt-forming mixture contains (R)2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3 -yl)acetic acid and 2-propanol in a weight ratio of from about 1.0:6.0 to about 1.0:8.0; and/or iii) the molar ratio of ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid and L-arginine is from about 1.0:0.93 to about 1.0:1.01; and/or iv) the weight ratio of L-arginine to H2O is from about 1.0:1.2 to about 1.0:1.5; and/or v) the temperature of said salt-forming mixture before adding said L-arginine is from 18°C to 30°C; and/or vi) said salt-forming mixture contains ^)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid acid, 2-propanol and water in a mass ratio of from about 1.0:6.0:0.25 to about 1.0:8.0:0.7; and/or vii) the temperature of said salt-forming mixture before adding said L-arginine is from 18°C to 30°C. 14. The method according to any one of claims 11 to 13, wherein said first additional amount of said water-miscible antisolvent is added at the first time and at the second time; optional where: i) from about 8% to about 12% of said first additional amount of said water-miscible anti-solvent is added at a first time point; and/or ii) said first additional amount of said water-miscible anti-solvent is added at said first time to obtain a cloudy mixture; and/or iii) before said second time point, optionally adding a seed crystal of L-arginine salt (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopent[b] indol-3-yl)acetic acid; and/or iv) said first additional amount of said water-miscible anti-solvent is added at said second time at such a rate as to complete the addition in about 1.00 hour or more. -