HK1135961B - Crystalline potassium salt of lipoxin a4 analogs - Google Patents

Description

Lipoxin A4Crystalline potassium salts of analogs

Technical Field

The present invention relates to lipoxin A₄Crystalline potassium of the analogSalts, their use for treating disease states characterized by inflammation, and pharmaceutical compositions comprising crystalline potassium salts of the analogs and processes for their preparation.

Background

Lipoxins, along with leukotrienes, prostaglandins, and thrombolectins, constitute a group of biologically active oxidized fatty acids collectively known as eicosanoids. Eicosanoic acid is synthesized de novo from membrane phospholipids via the arachidonic acid cascade of enzymes. Lipoxins of the eicosanoid type, which have been structurally unique, have been found to have potent anti-inflammatory properties since their initial discovery in 1984, suggesting that they may have therapeutic potential (Serhan, C.N., Prostagladins (1997), Vol.53, pp.107-137; O' Meara, Y.M. et al, Kidney Int. (Suppl.) (1997), Vol.58, pp.S56-S61; Brady, H.R. et al, curr.Opin. Nephrol. hypertens. (1996), Vol.5, pp.20-27; and Serhan, C.N., biochem. Biophys. acta. (1994), Vol.1212, pp.1-25). Of particular value is the ability of lipoxins to antagonize the pro-inflammatory functions of leukotrienes in addition to other inflammatory agents such as platelet activating factor, fMLP (formyl-Met-Leu-Phe) peptides, immune complexes, and TNF α. Thus, lipoxins are potent anti-neutrophil agents that inhibit Polymorphaneutrophil (PMN) chemotaxis, homotypic aggregation (homotypic aggregation), attachment, migration across endothelial and epithelial cells, margination (margination)/blood cell extravasation and tissue infiltration (Lee, T.H., et al, Clin.Sci. (1989), Vol.77, pp.195-203; Fiore, S., et al, Biochemistry (1995), Vol.34, pp.78-16686; Payiani, A., et al, J.Immunol. (1996), Vol.572, pp.2264-2272; Hedqvist, P., et al, acta.Physiol.157 and Sc. (1989), Vol.137, pp.157-pp.; Payiani, A, Intneykl, Kidnel et al, 1995-1295). Furthermore, lipoxins down-regulate the expression of endothelial P-selectin and PMN adhesion (Papyianni, A., et al, J.Immunol. (1996), Vol.56, pp.2264-2272), bronchial and vascular smooth muscle contraction, mesangial cell contraction and adhesion (Dahlen, S.E., et al, adv.Exp.Med.biol. (1988), Vo..229, pp.107-130; Chrise, P.E., et al, am.Rev.Respir.Dis. (1992), Vol.145, pp.1-1284; Badr, K.F., et al, Proc.Natl.Acad.Sci. (1989), Vol.86, 3438-3442; dy, Brambr.R., Amioj.J.1990, Phys.809, Phy.1990, Vol.O.259, Vol.49, Vol.F., pp.27, Vorgo.49, Vol.F., Australi.42, Vol.14, Vol.O.J.1990, Vol.O.J.259, Vol.O.18, Vorgo.49, Vol.F., and Haemophil.

Lipoxins, especially lipoxin A₄This unique anti-inflammatory property of (a) has led to an interest in exploring their use for the treatment of inflammatory or autoimmune diseases and pulmonary and respiratory tract inflammation. Such diseases and inflammations that exhibit significant inflammatory infiltration are particularly important, including, but not limited to, inflammatory bowel diseases such as Crohn's disease, skin diseases such as psoriasis, rheumatoid arthritis, and respiratory diseases such as asthma.

Like other endogenous eicosanoids, naturally occurring lipoxins are unstable products that are rapidly metabolized and inactivated (Serhan, C.N., Prostagladins (1997), Vol.53, pp.107-137). This has limited the development of research in the lipoxin field, particularly with respect to the in vivo pharmacological assessment of the anti-inflammatory properties of lipoxins. Several relate to the use of lipoxin A₄U.S. patents for compounds with active sites, but with longer tissue half-lives, have issued. See, e.g., U.S. patent nos. 5,441,951 and 5,648,512. These compounds retain lipoxin A₄Binding activity of receptors and potency of natural lipoxins in vitro and in vivo anti-inflammatory properties (Takano, T., et al, J.Clin.Invest. (1998), Vol.101, pp.819-826; Scalia, R., et al, Proc.Natl.Acad.Sci. (1997), Vol.94, pp.9967-9972; Takano, T., et al, J.exp.Med. (1997), Vol.185, pp.16993-1704; Maddox, J.F., et al, J.biol.chem. (1997), Vol.272, pp.6972-6978; Serhan, C.N., et al, Biochemistry (1995), Vol.34, pp.14609-14615).

Lipoxin A of interest to the present invention₄Analogs are disclosed in U.S. patent No. 6,831,186 and U.S. patent application No. 2004/0162433.

Those skilled in the art recognize crystalline, rather than amorphous, solidsDrug substances are particularly advantageous. The purified crystalline product is typically obtained during the formation of a crystalline solid by crystallization from a solution. Crystalline solid state forms are well characterized and generally exhibit greater stability than amorphous phases. By using crystalline solids as a component of a drug substance or drug, potential recrystallization of the amorphous phase, including changes in the properties of the drug substance or drug, can be avoided. Thus, there are lipoxin A disclosed in U.S. patent No. 6,831,186 and U.S. patent application publication No. 2004/0162433₄The need for stable crystalline solid state forms of the analogs.

Disclosure of Invention

The present invention relates to potent, selectively, and metabolically and chemically stable lipoxin A₄Crystalline potassium salts of analogs and their use in treating disease states characterized by inflammation, such as inflammatory or autoimmune diseases and pulmonary or respiratory inflammation, in mammals, particularly humans.

Accordingly, one aspect of the present invention relates to lipoxin A of formula (I)₄Crystalline potassium salt of the analog:

wherein:

R¹is-O-, -S (O)_t- (wherein t is 0, 1 or 2) or a straight or branched alkylene chain; and

R²is aryl (optionally substituted with one or more substituents selected from the group consisting of alkyl, alkoxy, halo, haloalkyl and haloalkoxy) or aralkyl (optionally substituted with one or more substituents selected from the group consisting of alkyl, alkoxy, halo, haloalkyl and haloalkoxy);

and wherein the compound of formula (I) is a single stereoisomer or a mixture of any of the stereoisomers.

The present invention includes all crystalline forms of the potassium salt of formula (I).

The salts of the invention show excellent stability under stress test conditions.

In another aspect, the invention relates to a process for preparing a crystalline potassium salt of formula (I), the process comprising I) co-mixing a potassium base in a suitable solvent and a free acid corresponding to the potassium salt of formula (I) in a suitable solvent; ii) optionally cooling the resulting suspension; iii) separating the resulting crystals from the resulting suspension; iv) optionally washing the isolated crystals with a suitable solvent; and v) drying the separated crystals to obtain crystalline potassium salt.

In one embodiment the process of the invention comprises preparing a suitable solution of an acid and a base, i.e. adding a suitable amount of water to induce the formation of crystals with high crystallinity.

In one embodiment the process of the invention comprises controlled drying of the salt to obtain the desired hydrate state, which is the dihydrate, monohydrate or dehydrated state.

In one embodiment the process of the present invention involves the preparation of a crystalline anhydrate (anhydrate) form of the potassium salt of formula (I) which comprises digesting (digesting) the hydrate or mixture of hydrates of potassium salt (I) in a suitable solvent.

Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a crystalline potassium salt of formula (I) as described above and one or more pharmaceutically acceptable excipients.

In another aspect, the invention relates to the use of a crystalline potassium salt of formula (I) as described above in the manufacture of a medicament for treating a mammal suffering from a condition characterized by inflammation, such as an inflammatory or autoimmune disease or inflammation of the lungs or respiratory tract.

In another aspect, the present invention relates to a method of treating a disease state characterized by inflammation in a mammal, particularly a human, wherein said method comprises administering to a mammal in need thereof a therapeutically effective amount of a crystalline potassium salt of formula (I) as described above. The disease state may be, for example, an inflammatory or autoimmune disease or inflammation of the lungs or respiratory tract.

Detailed Description

A. Definition of

All documents cited herein, including U.S. patents, U.S. patent application publications, and journal articles, are incorporated by reference herein in their entirety.

As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, "a" compounds refer to one or more of the above-mentioned compounds, while "the" enzyme "includes the particular enzyme as well as other family members and equivalents thereof (equivalents) known to those skilled in the art.

Unless otherwise specified, volume percentages are used herein.

Furthermore, as used in the specification and the appended claims, the following terms have the meanings indicated, unless otherwise specified to the contrary:

"alkyl" means a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, which is free of unsaturation, has one to eight carbon atoms, and which is attached to the remainder of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1-dimethylethyl (tert-butyl), and the like.

"alkylene chain" means a straight or branched divalent hydrocarbon chain consisting only of carbon and hydrogen, which does not contain unsaturation and has one to eight carbon atoms, such as methylene, ethylene, propylene, n-butylene, and the like.

"alkoxy" means a group of the formula-OR_aWherein R is_aIs an alkyl group as defined above.

"aryl" refers to a phenyl or naphthyl group. Unless otherwise specified, an aryl group may be optionally substituted with one or more substituents selected from alkyl, alkoxy (alkoy), halogen, haloalkyl or haloalkoxy. Unless otherwise specifically stated in the specification, it is to be understood that the above substitution may occur on any carbon atom of the aryl group.

"aralkyl" means a group of the formula-R_aR_bWherein R is_aIs an alkyl group as defined above and R_bIs an aryl group as defined above, such as benzyl and the like. The aryl group may be optionally substituted as described above.

"halogen" means bromine, chlorine, iodine or fluorine.

"haloalkyl" refers to an alkyl group as defined above substituted with one or more halo groups as defined above, for example trifluoromethyl, difluoromethyl, trichloromethyl, 2, 2, 2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl (1, 3-difluoroisopropyl), 3-bromo-2-fluoropropyl, 1-bromomethyl-2-bromoethyl (1, 3-dibromoisopropyl), and the like.

"haloalkoxy" means a compound of the formula-OR_cWherein R is_cAre haloalkyl groups as defined above such as trifluoromethoxy, difluoromethoxy, trichloromethoxy, 2, 2, 2-trifluoroethoxy, 1-fluoromethyl-2-fluoroethoxy, 3-bromo-2-fluoropropoxy, 1-bromomethyl-2-bromoethoxy and the like.

As used herein, "commercially available" compounds are available from standard Chemical supplies and other commercial sources, including, but not limited to, Acros Organics (Pittsburgh PA), Aldrich Chemical (Milwaukee WI, including Sigma Chemical and Fluka), apex Chemical Ltd. (Milton Park UK), Avoca Research (Lancashire U.K.), BDH Inc. (Canada), Bionet (Cornwall, U.K.), Chemservie Inc. (West Chemicals), CreScent Chemical Co., Hauppauge NY), Eastorganic Chemicals, Eastman Kodak Company TX (Rochester NY), Fisher Scientific Co., Pittsburgh Chemical, Inc. (Hipponship), Australin Chemical, Inc. (Rockwell Co., Inc.), and Australine Chemical Co., Australine Co., Inc. (Rockwell Co., Inc., where U., Riedel de Haenc AG (Hannover, Germany), Spectrum Quality Product, Inc. (New Brunswick, NJ), TCI America (Portland OR), Trans World Chemicals, Inc. (Rockville MD), and Wako Chemicals USA, Inc. (Richmond VA).

"mammal" includes humans and domestic animals such as cats, dogs, pigs, cattle, sheep, goats, horses, rabbits, and the like.

As used herein, "methods known to those skilled in the art" can be determined by various reference books and databases. Suitable reference books and treatises which specify the synthesis of the reactants used for preparing the compounds of the invention or which provide references to articles describing said preparation include, for example, "synthetic organic Chemistry", John Wiley&Sons, inc., New York; sandler et al, "Organic Functional Group Preparations," 2nd Ed., Academic Press, New York, 1983; H.O.House, "Modern Synthetic Reactions", 2nd Ed., W.A.Benjamin, Inc.Menlo Park, Calif.1972; gilchrist, "Heterocyclic Chemistry", 2nd Ed., John Wiley&Sons, New York, 1992; march, "Advanced Organic Chemistry: reactions, Mechanisms and Structure ", 4th Ed., Wiley-Interscience, New York, 1992. Specific and similar reactants are also identified by known Chemical catalogues written by the American Chemical abstracts of the American Chemical Society, which are available in most public and university libraries, and by online databases (the American Chemical Society, Washington, D).C, can be combined withwww.acs.orgContact for more detailed information). Chemicals known in the catalog but not commercially available can be prepared by custom chemical synthesis facilities, where many standard chemical supply facilities (such as those listed above) provide custom synthesis services.

"optional" or "optionally" or "may" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the described event or circumstance occurs and instances where it does not. For example, "optionally substituted aryl" means that the aryl group may or may not be substituted, and that this recitation includes both substituted aryl groups and unsubstituted aryl groups.

"polymorphs" refer to the various crystalline states of the same chemical compound of the present invention. Where "chemical compound" includes a molecule or ion (salt) or a mixture of a molecule and an ion (salt). The solid is present in amorphous or crystalline form. In the case of crystalline forms, the molecules are located systematically in 3-dimensional lattice sites. When a compound crystallizes from a solution or slurry, it may crystallize in different spatial lattice arrangements, a property known as "polymorphism", while different crystal forms are respectively referred to as "polymorphs". Different polymorphs of a given substance may differ from each other in one or more physical properties, such as solubility and dissolution, true density, crystal shape, compaction behavior, flow properties, and/or solid state stability. For chemicals that exist in two (or more) polymorphic forms, the form that is unstable at a given temperature is generally converted to the thermodynamically more stable form within a sufficient time. The thermodynamically unstable form is referred to as a "metastable" form. The conversion to a more stable form may be slow enough to assess the nature of the form and even suitable for pharmaceutical applications. Thus, it may be found that the metastable form exhibits sufficient chemical and physical stability under normal storage conditions to allow its use in commercial form. In this case, the metastable forms, although less stable, may exhibit more favorable properties than those of the stable forms, such as increased solubility or better oral bioavailability.

"solvate" refers to an aggregate comprising one or more molecules of the invention and a non-stoichiometric amount of one or more solvent molecules or solvents. The solvent may be water, in which case the solvate is referred to as a hydrate. Alternatively, the solvent may be an organic solvent. Thus, lipoxin A of formula (I)₄The potassium salts of the analogs may exist as hydrates, including the monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate, dehydrated hydrates having their non-stoichiometric water content, and the like, as well as the corresponding solvated forms. The potassium salt of formula (I) may be a true solvate, while in other cases the salt may simply be water that remains adventitious (adhentitous) or a mixture of water plus some adventitious solvent.

See, e.g., Byrn, S et al, "Solid State Chemistry of Drugs", SSCI (1999), wherein polymorphs and solvates, their features and properties, and associations with drug substances and drug products are discussed; and Stahl, P and Wermuth, C "Handbook of pharmaceutical salts", Wiley (2002), where salts, their preparation and properties are discussed.

As used herein, "suitable conditions" for carrying out a synthetic step are provided explicitly herein or can be determined by reference to publications relating to methods for synthesizing organic chemistry. The above references and papers detail the synthesis of reactants used in the preparation of the compounds of the invention and will also provide suitable conditions for carrying out the synthetic steps of the invention.

By "suitable solvent" is meant any solvent that is compatible with the reaction components and the reaction conditions. The term encompasses a solvent or mixture of solvents and includes, but is not limited to, organic solvents and water. Suitable solvents are known to those skilled in the art or can be determined by reference to publications relating to methods used in synthetic organic chemistry.

By "therapeutically effective amount" is meant an amount of the potassium salt of the present invention that is sufficient to treat a disease state characterized by inflammation as described below when administered to a mammal, particularly a human, in need thereof. The amount of the potassium salt of the present invention which constitutes a "therapeutically effective amount" will vary depending on the salt, its solvate form, the disease state to be treated and its severity, the age of the mammal being treated, etc., but can be routinely determined by those skilled in the art.

As used herein, "treating" or treatment "includes treating a disease state characterized by inflammation, such as an inflammatory or autoimmune disease or pulmonary or respiratory inflammation, in a mammal, particularly a human, and includes:

(i) preventing the disease or inflammation from occurring in a mammal, particularly a human, particularly when the mammal is predisposed to the disease but has not been diagnosed as having the disease;

(ii) inhibiting, i.e. arresting the development of, said disease or inflammation; or

(iii) Alleviating the disease or inflammation, i.e., causing inflammation or disease regression.

The potassium salts of formula (I) have three asymmetric centers and thus exist as enantiomers, diastereomers, and other stereoisomeric forms defined as (R) -or (S) -or as (D) -or (L) -depending on the absolute stereoconfiguration. The present invention is intended to include all such possible isomers, as well as their racemic and optically pure forms. The optically active (+) and (-), (R) -and (S) -, or (D) -and (L) -isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques such as HPLC using a chiral column. The compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and are intended to include both E and Z geometric isomers unless otherwise specified. Likewise, all tautomeric forms are also intended to be included.

"stereoisomers" refers to compounds that are made up of the same atoms joined by the same bonds but have different three-dimensional structures, which are not interchangeable. The present invention contemplates various stereoisomers and mixtures thereof, and includes "enantiomers" which refer to two stereoisomers whose molecules are nonsuperimposable mirror images of each other.

B. Crystalline potassium salt of the invention

Salt formation is known to significantly increase the stability of the drug relative to its free acid, and the stability of different salts of the salt-forming agent varies. It is also known that crystalline salt forms are more stable than amorphous salt forms.

Thus, a study was conducted aimed at finding suitable stable crystalline salts of the acids of formula (II):

wherein:

R¹is-O-, -S (O)_t- (wherein t is 0, 1 or 2) or a straight or branched alkylene chain; and

R²is aryl (optionally substituted with one or more substituents selected from the group consisting of alkyl, alkoxy, halo, haloalkyl and haloalkoxy) or aralkyl (optionally substituted with one or more substituents selected from the group consisting of alkyl, alkoxy, halo, haloalkyl and haloalkoxy);

and wherein the acid of formula (II) is a single stereoisomer or a mixture of any of the stereoisomers.

By salt screening tests with various organic and inorganic cations, it was very surprisingly found that the crystalline potassium salt of the acid (IIa) is significantly more stable than all other salts prepared and more stable than its acid (IIa).

In one embodiment, the present invention relates to a crystalline potassium salt of formula (I) as disclosed and defined in the summary of the invention section.

In another embodiment, the invention relates to a crystalline potassium salt of formula (I), wherein R¹is-O-and R²Is phenyl optionally substituted with one or more substituents selected from fluoro, chloro and iodo.

In another embodiment, the invention is a crystalline potassium salt of formula (I) wherein R is¹is-O-and R²Is 4-fluorophenyl.

In another embodiment, the compounds of the invention are selected from the following compounds:

potassium 2- ((2S,3R,4E,6E,10E,12S) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetate;

potassium 2- ((2R, 3R,4E,6E,10E,12S) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetate;

potassium 2- ((2S, 3S, 4E,6E,10E,12S) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetate;

potassium 2- ((2R, 3S, 4E,6E,10E,12S) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetate;

potassium 2- ((2S,3R,4E,6E,10E, 12R) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetate;

potassium 2- ((2R, 3R,4E,6E,10E, 12R) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetate;

potassium 2- ((2S, 3S, 4E,6E,10E, 12R) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetate; and

potassium 2- ((2R, 3S, 4E,6E,10E, 12R) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetate.

In another aspect, the invention relates to a crystalline potassium salt, wherein the crystalline potassium salt is potassium 2- ((2S,3R,4E,6E,10E,12S) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetate (Ia):

C. salt screening:

the salt screening assay is described in detail below:

materials:

a number of inorganic cations were investigated to determine their ability to form stable crystalline salts of acid (IIa), including potassium, sodium, calcium, aluminum, magnesium, lithium and zinc.

Several organic amines were studied, including N-methylglucamine, diethanolamine, ethylenediamine, choline, lysine and arginine.

In addition to the foregoing, formation of mixed salts (mixed salts) of acids of formula (IIa) with polyvalent inorganic cations and organic acids has also been investigated.

Test parameters are as follows:

typically, salt screening assays involve variation of the following different assay parameters:

i) the type of cationic compound used (e.g. carbonate, hydroxide, methoxide, ethoxide).

ii) the type of solvent used.

iii) the exclusive addition or removal of water (to drive hydrate or anhydrate formation).

iv) the type of crystallization used (cooling crystallization, anti-solvent crystallization, or evaporative crystallization, and combinations thereof).

v) type of screening (manual as well as high throughput screening).

As a result:

it was surprisingly found that it is only possible to form a suspension and isolate the crystalline salt of the acid (IIa) when using a potassium compound as base. Tests with other cations produced solutions with no solid phase, only an oil phase, or amorphous solids. The formation of mixed salts also failed. Similarly, high throughput screening did not yield other crystalline salts of acid (IIa).

D. Solid state forms of potassium salts of formula (Ia)

The crystalline potassium salt of formula (Ia) forms three well-defined crystalline forms: anhydrate and two hydrates. In addition, two dehydrated hydrate forms (dehydrated form I and dehydrated form II) were obtained, and the amorphous phase was detected in some studies. No solvate was detected. The present invention encompasses all crystalline forms of the potassium salts of formula (I) and (Ia).

The different solid state forms of the potassium salt (Ia) can be distinguished by the position of the reflected peaks in the X-ray powder diffraction (XRPD) pattern. The strongest reflections for the anhydrate, monohydrate and dihydrate of crystalline potassium salt (Ia) are shown in table 1:

TABLE 1

X-ray powder diffraction data for anhydrate, monohydrate, and dihydrate-d-value of strongest reflection, 2 Θ -value (germanium-monochromated CuK α)₁-radiation)

An anhydrate form of the crystalline potassium salt of formula (Ia) at about d ═ 26.7And d is 3.7Characteristic peaks are shown.

The monohydrate form of the crystalline potassium salt of formula (Ia) is at about d-26.5About d-4.3And about d 4.0Shows characteristic peaks, and the dihydrate form of the crystalline potassium salt of formula (Ia) is at about d-27.1About d-4.2And about d 4.1Characteristic peaks are shown.

The XRPD patterns of the dehydrated form I and dehydrated form II of the crystalline potassium salt of formula (Ia) show that the peak positions differ slightly between batches of the two dehydrated forms due to variations in solvent residues. Therefore, the exact d-value, 2 Θ -value and relative intensity characterizing both forms cannot be given. However, they are at about d-24.6And about 27.1Characteristic peaks are shown in the middle.

It will be appreciated by those skilled in the art that the obtained X-ray diffraction patterns may have measurement errors depending on the applied measurement conditions. In particular, it is well known that the intensity of an X-ray diffraction pattern can fluctuate with the crystal habit of a substance and the measurement conditions applied. It will also be appreciated that the relative intensities also vary with the test conditions, and therefore the exact intensity level should not be considered. Furthermore, the measurement error of the diffraction angle θ of a conventional X-ray diffraction pattern at a given temperature is generally about. + -. 0.1, which should be taken into account when referring to the aforementioned diffraction angles. Thus, reference to an X-ray powder diffraction pattern when the term "about" is used herein means that the crystalline form of the present invention is not limited to a crystalline form that provides an X-ray diffraction pattern that is substantially identical to the X-ray diffraction pattern described in the figures disclosed herein. Any crystalline form that provides an X-ray diffraction pattern substantially in accordance with those disclosed in the accompanying drawings is within the scope of the invention. Even if the X-ray diffraction patterns are not completely consistent, the ability to determine whether the polymorphic forms of a compound are the same is within the purview of one skilled in the art.

IR-spectra of crystalline Potassium salt (Ia) hydrates showed them to be 1214cm^-1And 1127cm^-1With a characteristic IR band.

The water content of the crystalline potassium salt (Ia) determines its solid state form (anhydrate, monohydrate, dihydrate, dehydrated form I, or dehydrated form II). The actual water content of the salt is determined by the ambient relative humidity and temperature.

The interconversion of the monohydrate and dihydrate and dehydrated form II is reversible. The anhydrate is converted to the dihydrate via the monohydrate. This conversion is irreversible. The dihydrate is converted to the dehydrated form II by dehydration. The dehydrated form I is converted to the monohydrate and the dihydrate. No conversion of the monohydrate or dihydrate form to the dehydrated form I was observed.

The conversion of the monohydrate to the dihydrate form and vice versa occurs rapidly and is clearly defined at 25 ℃ and 40 ℃. This is shown by dynamic moisture sorption studies. For example, during the pre-drying of the monohydrate at 25 ℃ and 0% relative humidity, 4.0% of water is released from the monohydrate. This results in the formation of dehydrated form II. Between 20% and 40% relative humidity, one mole of water is absorbed resulting in the formation of a monohydrate. Until the relative humidity is about 50% monohydrate is stable, and then converted to the dihydrate by absorption of a second mole of water at a relative humidity between 50% and 60%. The dihydrate was in a stable form at 25 ℃ up to a relative humidity of 98%. In the 40% relative humidity and 50% desorption cycles, conversion to the monohydrate occurred. The monohydrate formed is present up to a relative humidity of 10%. Dehydration of the monohydrate to dehydrated form II occurred between 0% and 10% relative humidity. Figure 1 shows the corresponding adsorption isotherms. No significant difference was observed between the two cycles.

The hygroscopicity with respect to the conversion of the monohydrate and the dihydrate was re-investigated between relative humidities of 45% and 55% with a smaller step of relative humidity. This study showed that the equilibrium state of the two hydrates at 25 ℃ was between 48% and 50% relative humidity. The reversible conversion between monohydrate and dihydrate at 40 ℃ is between 57% and 59% relative humidity, about 10% higher than at 25 ℃. A more detailed study of the desorption range of monohydrate at 40 ℃ between 0% and 10% relative humidity showed that desorption occurred between 2% and 3% relative humidity.

Likewise, dynamic moisture sorption studies at 25 ℃ showed the conversion of the anhydrous to hydrate form of the potassium salt (Ia). Until a relative humidity of 40% anhydrous is stable. Absorption of about two moles of water at between 40% and 60% relative humidity results in the formation of dihydrate.

Data collection for XRPD in transmission mode using germanium-monochromatization cuka at room temperature (between 20 ℃ and 25 ℃)₁-radiation (λ ═ 1.5406)) The process is carried out. By operating the X-ray tube with a copper anode at 40kV and 30 mA. If an orifice plate is used, a small linear position sensitive detector (small linear position sensitive detector) is used to provide angular resolution of 0.08 ° between 3 ° ≦ 2 Θ ≦ 35 ° and 2 ° ≦ 2 Θ ≦ 35 ° (step width 0.5 °), or between 7 ° ≦ 2 Θ ≦ 35 ° (step width 0.5 °)Line 2 Θ scans. The sample was enclosed between two sheets of polyester film or two sheets of aluminum foil bonded by double-sided tape to avoid the effect of humidity during the measurement. Data acquisition and evaluation Using STOEWinX^powThe software package is run.

The IR spectra were recorded using diffuse reflectance techniques. The sample was diluted with potassium bromide in a ratio of 1: 100.

Data collection for the hygroscopicity experiments was run on an automatic moisture sorption analyzer. The investigated solid state form of about 20mg of crystalline potassium salt (Ia) was exposed to a continuous stream of nitrogen at a predetermined and constant relative humidity. The velocity of sweep gas (sweep gas) was set at 200cm³And/min. Two complete cycles (adsorption/desorption) were measured at 25 ℃ for the basic analysis. The above measurement starts from a relative humidity of 0% in order to remove moisture from the surface. Once constant weight is reached, the next humidity is automatically set. Moisture adsorption/desorption at relative humidity between 0% and 90% was studied at 10% step size under the first step initiation criteria as described herein. In addition, adsorption at a relative humidity of about 98% was investigated. Data were collected using DVSWin software. Moreover, a study at 40 ℃ was carried out and, in order to evaluate the monohydrate/dihydrate equilibrium in more detail, the step size of the relevant area was shortened (each step equals 1% relative humidity).

E. Chemical stability of solid state form of potassium salt (Ia)

The chemical stability of the anhydrate and monohydrate of the potassium salt of formula (Ia) of the invention and of the dehydrated form I of potassium salt (Ia) of the invention was assessed by comparing the stability at 40 deg.C/75% rH over a 4 week period. The results are shown in table 2 below.

TABLE 2

^*Starting with the anhydrate during the test, most likely the dihydrate form of the potassium salt (Ia) at the end of the test

^**Starting with the monohydrate during the test, most likely in the form of the dihydrate of the potassium salt (Ia) at the end of the test

The potassium salt (Ia) is sufficiently stable in all three forms; however, the anhydrate and monohydrate of the potassium salt (Ia) showed no signs of degradation.

F. Preparation of Potassium salts of formulae (I) and (Ia)

The compounds of formula (II), i.e., the acids, are described in detail in U.S. patent No. 6,831,186 and U.S. patent application No. 2004/0162433, the disclosures of which are incorporated herein by reference in their entirety.

The process for preparing the crystalline potassium salt is illustrated by using the salt of formula (Ia), but it can be used to prepare crystalline potassium salts of other compounds of formula (I). In general, the process for preparing crystalline potassium salts (Ia) is based on the neutralization of the acid (IIa) by treatment with a base. The resulting mixture forms a suspension, which is then optionally cooled. The crystals are separated from the suspension and dried to yield the desired hydrate form of the crystalline potassium salt (Ia).

More specifically, the acid (IIa) is dissolved in a suitable solvent such as ethanol. A potash such as potassium methoxide or potassium hydroxide is then dissolved in a suitable solvent such as ethanol. A solution of potash is added to a solution of acid (IIa) and vice versa.

The dosage of the potassium base relative to the solution of acid (III) is controlled by measuring the pH of the resulting mixture in a manner well known to those skilled in the art, for example, with a wetted pH-indicating dipstick. In one embodiment, the addition of the potash solution is typically stopped when a pH of 6 to 15 is reached, typically a pH of 7 to 14, in a currently preferred embodiment, when a pH of 8 to 13 is reached.

The dehydrated form I of potassium salt (Ia) is obtained by removing water from the reaction mixture. This can be achieved, for example, by using an anhydrous solvent and forming the salt by adding potassium methoxide.

Furthermore, the dehydrated form I of the potassium salt (Ia) can be obtained by suspending the monohydrate form of the crystalline potassium salt (Ia) in an organic solvent such as acetonitrile. No direct conversion of the monohydrate form or the dihydrate form to the dehydrated form I was observed by drying or heat treatment.

To avoid obtaining the now less desirable dehydrated hydrate form (I) of potassium salt (Ia) in the crystallization step, the solvent of crystallization contains at least some water. Typically, the final reaction mixture comprises at least about 0.01% water, typically from about 0.5% to about 99.9% water, and in one presently preferred embodiment, from about 1% to about 30% water. Thus, for example, in one embodiment, the acid (IIa) -solvent solution should contain at least some water. Typically, the acid (IIa) -solvent solution contains at least about 0.01% water, typically from about 0.5% to about 99.9% water, and in one presently preferred embodiment, from about 1% to about 30% water. Also, in one embodiment, the potash-solvent solution should contain at least some water. Typically, the potash-solvent solution comprises at least about 0.01% water, typically from about 0.5% to about 99.9% water, and in one presently preferred embodiment, from about 1% to about 30% water. The resulting reaction mixture will contain at least some water. The resulting mixture forms a suspension, optionally cooled before the steps of solid/liquid separation and optionally washing with a suitable solvent. Typically, the wash solvent comprises at least about 0.01% water, typically from about 0.5% to about 99.9% water, and in one presently preferred embodiment, from about 1% to about 30% water. Followed by a drying step.

The method of drying the isolated crystals defines the hydrate form (monohydrate or dihydrate or dehydrated hydrate) of the crystalline potassium salt produced, and therefore should be suitably controlled when it is desired to obtain the mono-or dihydrate of the potassium salt (Ia). Also, the drying process avoids the formation of the less desirable dehydrated form (II) of potassium salt (Ia), which is obtained by overdrying the product. The following section G describes the optimization of the above method.

The dehydrated form II of potassium salt (Ia) may also be formed by dehydration of the hydrate by heat treatment (typically at temperatures below 100 ℃) or storage at 0% relative humidity. Dehydration of the monohydrate dihydrate gives the dehydrated form II.

In general, the process for preparing the anhydrous form of the potassium salt (Ia) is based on cooking the hydrated form of the potassium salt in a suitable solvent at a higher temperature for a period of time. The crystals were separated from the suspension and then dried to give the potassium salt (Ia) as an anhydrate.

More specifically, the hydrate of the potassium salt is digested in a suitable solvent such as 1, 4-dioxane. The suspension is stirred and heated to almost the boiling point of the solvent, usually for about 0.1 to about 10 hours, generally about 1 to about 8 hours, preferably about 2 to about 6 hours. The suspension is then cooled, the crystals are isolated, optionally washed with a suitable solvent such as 1, 4-dioxane, and the crystals are subsequently dried to give the potassium salt (Ia) in the anhydrous form.

G. Optimization of drying method of crystal potassium salt of formula (Ia)

The formation of the preferred crystalline hydrate form of potassium salt (Ia) is optimized by varying the drying conditions.

XRPD analysis of the wet cake revealed that the dihydrate form of the potassium salt formed during the crystallization step. The filter cake (i.e., the dihydrate form of the potassium salt) loses moisture during drying. But when the dihydrate form of the potassium salt loses its moisture and converts to the monohydrate form, the monohydrate form may then continue to convert to the less desirable dehydrated form II.

As is evident from the DVS data, the anhydrate-monohydrate conversion is rapid and conventional drying methods such as passing through a vacuum drying chamber do not produce dihydrate. There is also a potential risk of overdrying the monohydrate to dehydrated form II.

Thus, the drying process is optimized to produce either the dihydrate or the monohydrate and to safely avoid over-drying to produce dehydrated form II.

Optimized drying process with well regulated and controlled through-air flow(sweet gas). By specially controlling and setting the sweep gas (e.g. N)₂) And the pressure and temperature within the dryer, under temperature and humidity conditions associated with a stability domain of the desired hydrate form. According to this process, a specific hydrate form of the potassium salt (Ia) can be obtained.

Also, studies have shown that the use of a sweep gas is advantageous in ensuring adequate drying times and avoiding condensation of the liquid in the dryer. In addition to the sweep gases described above, any suitable sweep gas may be used in the present invention, which are known to those skilled in the art or which may be determined without undue experimentation.

In addition to the use of a vacuum drying chamber, any drying method or apparatus may be used in the methods of the present invention for drying the potassium salts of formula (I) and (Ia), including, but not limited to, a hybrid dryer (e.g., a paddle dryer or a cone dryer), a contact dryer, a convection dryer, an infrared dryer, and the like, as are known or determinable to those skilled in the art.

H. Application of crystal sylvite of the invention

The crystalline potassium salt of formula (I) has a structure similar to natural lipoxin A₄But has greater resistance to chemical and metabolic degradation. Thus, the crystalline potassium salts of formula (I) are useful in the treatment of inflammatory or autoimmune diseases in mammals, particularly humans. In particular, the crystalline potassium salt of formula (I) is useful for inhibiting acute or chronic inflammation or inflammatory or autoimmune responses mediated by neutrophils, eosinophils, T lymphocytes, NK cells, or other immune cells that contribute to the pathogenesis of inflammatory, immune, or autoimmune diseases. The crystalline potassium salt of formula (I) may also be used to treat proliferative diseases including, but not limited to, those associated with inflammation or disorders in the immune response, such as cancer. The crystalline potassium salt of formula (I) is also useful as an inhibitor of the angiogenic response in the pathogenesis of cancer.

Thus, the crystalline potassium salts of formula (I) are useful in the treatment of the following inflammatory or autoimmune diseases in mammals, particularly humans: allergic reactions, allergic contact dermatitis, allergic rhinitis, chemical and non-specific irritant contact dermatitis, urticaria, atopic dermatitis, psoriasis, fistulas associated with crohn's disease, pouchitis (pouchitis), septic or endotoxic shock, hemorrhagic shock, shock-like syndrome, capillary leak syndrome induced by immunotherapy of cancer, acute respiratory distress syndrome, traumatic shock, immune-and pathogen-induced pneumonia, immune complex-mediated lung injury and chronic obstructive pulmonary disease, inflammatory bowel disease (including ulcerative colitis, crohn's disease and post-operative trauma), gastrointestinal ulcers, diseases associated with ischemia-reperfusion injury (including acute myocardial ischemia and infarction, acute renal failure, ischemic bowel disease and acute blood loss or ischemic stroke), Immune-complex-mediated glomerulonephritis, autoimmune diseases (including insulin-dependent diabetes, multiple sclerosis, rheumatoid arthritis, osteoarthritis and systemic lupus erythematosus), acute and chronic organ transplant rejection, transplant atherosclerosis and fibrosis, cardiovascular diseases (including hypertension, atherosclerosis, aneurysms, severe lower limb ischemia, peripheral arterial occlusive disease and raynaud's syndrome), diabetic complications (including diabetic nephropathy, neuropathy and retinopathy), diseases of the eye (including macular degeneration and glaucoma), neurodegenerative diseases (including stroke, alzheimer's disease, parkinson's disease, encephalitis and delayed neurodegeneration in HIV dementia), inflammatory and neuropathic pain including arthritic pain, periodontal disease including gingivitis, ear infections, autoimmune diseases (including insulin-dependent diabetes mellitus, multiple sclerosis, rheumatoid arthritis, osteoarthritis and systemic lupus erythematosus), inflammatory and neuropathic pain, Migraine, benign prostatic hyperplasia, cancers including but not limited to leukemia and lymphoma, prostate cancer, breast cancer, lung cancer, malignant melanoma, kidney cancer, head and neck tumors, and colorectal cancer.

The crystalline potassium salt of formula (I) is also useful for treating folliculitis induced by an Epidermal Growth Factor (EGF) or Epidermal Growth Factor Receptor (EGFR) kinase inhibitor for use in the treatment of solid tumors. Clinical trials have shown that folliculitis (inflammation of the hair follicles manifested as severe acneiform skin rash on the face, chest and upper back) is the major dose-limiting side effect of the above treatments. This folliculitis is associated with neutrophil infiltration, suggesting that products secreted by active neutrophils are responsible for inflammation. The crystalline potassium salts of formula (I) inhibit neutrophil-or eosinophil-mediated inflammation and are therefore useful in the treatment of such folliculitis, thereby improving the quality of life of patients receiving cancer treatment and allowing geigers of EGF inhibitors or EGFR kinase inhibitors or prolonging the treatment period, resulting in improved efficacy of the desired inhibitors.

The crystalline potassium salt of formula (I) may also be used to treat pulmonary and respiratory tract inflammation, including but not limited to asthma, chronic bronchitis, bronchiolitis obliterans (including bronchiolitis obliterans with accompanying pneumonia), respiratory allergic inflammation (including rhinitis and sinusitis), eosinophilic granuloma, pneumonia, pulmonary fibrosis, pulmonary manifestations of connective tissue disease, acute or chronic lung injury, chronic obstructive pulmonary disease, adult-type respiratory distress syndrome, and other noninfectious inflammatory diseases of the lungs characterized by eosinophilic infiltration. For example, a crystalline potassium salt of formula (I) can be used to inhibit: eosinophil-mediated inflammation of the lung or tissue; neutrophil-mediated lung inflammation; lymphocyte-mediated lung inflammation; production of cytokines and chemokines, including interleukin-5, interleukin-13, and eotaxin; production of lipid mediators (lipid mediators), including prostaglandin E₂And cysteinyl leukotrienes; airway hyperresponsiveness; and airway and vascular inflammation.

I. Test of crystalline Potassium salt of the invention

Markers of inflammation are the attachment and migration of neutrophils, eosinophils, and other inflammatory cells to the endothelium. Similar processes are observed that occur in the migration of cells in the lung, gastrointestinal tract and other organs between polarized epithelial cells. Cell culture models of these processes are available and have been used to demonstrate lipoxin A₄And stabilized lipoxin A₄The analogues inhibit the passage of human neutrophils across human endothelial and epithelial cells including the human gutEpithelial cell line T₈₄Shift of (2). Thus, one skilled in the art can readily prepare such compounds by performing procedures similar to those described in Colgan, S.P., et al, J.Clin.Invest, (1993), Vol.92, No.1, pp.75-82; and Serhan, C.N., et al, Biochemistry (1995), Vol.34, No.44, pp.14609-14615, to test the ability of crystalline potassium salts of formula (I) to inhibit migration of human neutrophils and eosinophils across human endothelial and epithelial cells.

The balloon model and/or the mouse zymosan-induced peritonitis model can be used to assess the in vivo efficacy of the crystalline potassium salt of formula (I) in treating inflammatory responses. These are acute experimental models of inflammation characterized by infiltration of inflammatory cells into a localized area. References are as described in Ajuebor, m.n., et al, Immunology (1998), vol.95, pp.625-630; gronert, k, et al, am.j.pathol (2001), vol.158, pp.3-9; pouliot, M., et al, Biochemistry (2000), Vol.39. pp.4761-4768; clish, c.b., et al, proc.natl.acad.sci.u.s.a. (1999), vol.96, pp.8247-8252; and in vivo assays in Hachicha, M., et al, J.Exp.Med. (1999), Vol.189, pp.1923-30.

Animal models (i.e., in vivo assays) may also be used to determine the efficacy of crystalline potassium salts of formula (I) in the treatment of asthma and lung and respiratory related diseases. See, e.g., those described in De sancts, G.T., et al, journal of Clinical Investigation (1999), Vol.103, pp.507-515; and Campbell, E.M., et al, J.Immunol. (1998), Vol.161, No.12, pp.7047-7053.

Alternatively, the effectiveness of the crystalline potassium salt of formula (I) for use in the claimed method can be tested by employing the methods described in U.S. patent No. 6,831,186 and U.S. patent application publication No. 2004/0162433, the relevant disclosures of which are fully incorporated herein in their entirety.

J. Administration of the crystalline Potassium salt of the invention

The crystalline potassium salt of formula (I) may be administered as a single stereoisomer or any mixture of stereoisomers, or as a cyclodextrin inclusion compound thereof, or as a solvate or polymorph thereof, in pure form or in an appropriate pharmaceutical composition by any acceptable mode of administration or as a medicament serving a similar application. Thus, administration can be, for example, orally, nasally, parenterally, pulmonarily, topically, transdermally, or rectally in solid, semi-solid, lyophilized powder, or liquid dosage forms, such as tablets, suppositories, pills, soft elastic and hard gelatin capsules, powders, solutions, suspensions, aerosols, patches, and the like, preferably in unit dosage forms suitable for the simple administration of precise dosages. The composition should comprise the potassium salt of the invention as (one of) the active drug(s) and conventional pharmaceutical carriers or excipients and may additionally comprise other medicinal agents, drugs, carriers, adjuvants and the like known to those skilled in the art.

Actual methods of preparing the above dosage forms are known or will be known to those skilled in the art, see, for exampleRemington′s Pharmaceutical Sciences18th Ed., (Mack Publishing Company, Easton, Pennsylvania, 1990). In any event, the composition to be administered should contain a therapeutically effective amount of the crystalline potassium salt of formula (I) for the treatment of a disease state characterized by inflammation, in accordance with the present disclosure.

Generally, depending on the intended mode of administration, the pharmaceutically acceptable composition will comprise from about 0.1% to about 99.9% by weight of the crystalline potassium salt of formula (I) and from about 99.9% to about 0.1% by weight of a suitable pharmaceutical excipient.

In one embodiment, the route of administration is oral, using a convenient daily regimen that can be adjusted according to the severity of the disease state being treated. For oral administration as described above, a pharmaceutically acceptable composition comprising the crystalline potassium salt of formula (I) is formed by the addition of one or more commonly employed pharmaceutically acceptable excipients. The composition is in the form of solution, suspension, tablet, pill, capsule, powder, sustained release preparation, etc.

Preferably, the above composition takes the form of a capsule, caplet or tablet and therefore also typically comprises diluents, disintegrants, lubricants and binders.

The crystalline potassium salts of formula (I) may also be formulated as suppositories containing the active ingredient placed in the carrier, which is slowly dissolved in the body, such as those typically employed for this ability.

Liquid pharmaceutically administrable compositions can be formulated, for example, by dissolving, dispersing, etc., the potassium salt of the present invention and optional pharmaceutically acceptable adjuvants in a carrier, such as water, saline, aqueous dextrose, glycerol, ethanol, and the like, to form a solution or suspension.

If desired, the pharmaceutical compositions of the present invention may also contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, antioxidants and the like.

The crystalline potassium salt of formula (I) is administered in a therapeutically effective amount, which will vary depending on a variety of factors, including the activity of the particular drug employed; metabolic stability and length of action of crystalline potassium salts of formula (I); the age, weight, overall health, sex, and diet of the patient; the mode and time of administration; the rate of excretion; combined medication; the severity of the particular disease state being treated; and a subject receiving treatment.

Examples

The following examples further describe the preparation of crystalline potassium salts of formula (Ia). Other lipoxins A of formula (I)₄Crystalline potassium salts of the analogs can also be prepared similarly.

Example 1:monohydrate forms of crystalline potassium salts (Ia) prepared by crystallization and drying under conditions associated with a monohydrate stability domain

2- ((2S,3R,4E,6E,10E,12S) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetic acid (compound (IIa)) (964g) was dissolved in a mixture of absolute ethanol (3.850L) and water (0.580L). In a second flask, potassium hydroxide pellets (172.2g) were dissolved in a mixture of water (0.580L) and absolute ethanol (3.850L). Potassium hydroxide solution was slowly added to the solution of the starting acid (IIa) at 20 ℃ until pH 8.5, measured using a wet pH paper. Seed crystals (2.5g) were added to the solution. After 90min at 22 ℃ a seed bed formed, the suspension was cooled to-10 ℃ within 45min and stirred at this temperature for a further 2 hours. The cooled suspension was then filtered using a suction filter with filter paper. The collected crystals were washed 3 times with 1.6L of a cooled ethanol/water (-10 ℃ C.; 6.0% by weight water) mixture and then dried to constant weight in a vacuum drying chamber at 200mbar/30 ℃ using air (relative humidity-30%) as a sweep gas. The drying process was controlled by measuring the moisture content (via a Karl moisture titrator) of several samples taken during the drying process. A crystalline monohydrate form (901g) of the potassium salt of formula (Ia) is obtained.

Example 2:preparation of the monohydrate form of the crystalline Potassium salt (Ia) by drying with a microwave dryer

2- ((2S,3R,4E,6E,10E,12S) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetic acid (compound (IIa)) (50.05g) was dissolved in a mixture of absolute ethanol (158.11g) and water (30.07 g). In a second flask, potassium hydroxide granules (12.24g) were dissolved in a mixture of water (41.22g) and absolute ethanol (216.15 g). Potassium hydroxide solution (180.21g) was slowly added to the solution of the starting acid (IIa) at 20 ℃ until pH 8.5, as measured using wet pH paper. The above solution was cooled to 0 ℃. The seeds were suspended in ethanol and the resulting suspension (0.3g) was added to the solution. After 30min, a seed bed formed, and the suspension was cooled to-12 ℃ over 30min and stirred at this temperature for a further 1 h. The cooled suspension was then filtered using a suction filter with filter paper. The collected crystals were washed with 0.1L of a cooled ethanol/water mixture (-12 ℃ C.; 4.0% by weight of water). Samples from the wet filter plate analyzed by XRPD showed dihydrate. The drying process then starts in the microwave dryer. Drying conditions are as follows: the power is 80W, and the pressure in the dryer is less than 150 mbar. The crystalline constant weight after 3.7 hours drying time gave the crystalline monohydrate form (38.97g) of the potassium salt of formula (Ia) (XRPD-analysis, thermogravimetric analysis).

Example 3: preparation of dihydrate forms of crystalline Potassium salt (Ia) by crystallization and drying under conditions associated with the dihydrate stability Domain

2- ((2S,3R,4E,6E,10E,12S) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetic acid (compound (IIa)) (500mg) was dissolved in ethanol (2 mL). Water (0.75mL) was added to the solution. The resulting clear solution was stirred. Potassium hydroxide granules (85.3mg) were dissolved in ethanol (2.84 mL). The resulting solution was clear and had no solid phase. An equimolar amount of a clear solution of potassium hydroxide is added to the solution of the starting acid (IIa). The dosage of potassium hydroxide addition was performed under pH validation (wet pH indicator strip). The addition of potassium hydroxide solution was stopped when the solution pH reached 8.5. The stirred solution was cooled to-10 ℃. After 2 hours, a suspension was found. The suspension was stirred for another 1 hour. The suspension was centrifuged in a pre-cooled centrifuge and the mother liquor decanted. The solid phase was dried in a circulating air dryer at 30 ℃. The circulating air dryer maintains a relative humidity of 70% by humidifying the permeate air. Through air flow (N)₂) Is continuously introduced into the dryer. The flow of permeate gas is alternately dry or wet with water. Wetting N₂Relative flow drying N₂The time period of the flow is adjusted by automatic control techniques taking into account the relative humidity within the dryer. Using this technique, it is possible to ensure a relative humidity of 70% in the dryer during the storage time. XRPD analysis of the wet filter pad and solid samples in the dryer after 8 hours and 32 hours drying time (═ dry product) showed the crystalline dihydrate form of the potassium salt (Ia).

Example 4:preparation of dihydrate form of crystalline Potassium salt (Ia) from monohydrate form by storage under dihydrate conditions

The monohydrate form of the potassium salt (Ia) (1.697g) was transferred to a dish. The monohydrate form of the substance was determined by XRPD-analysis and thermogravimetric analysis at the start of the process. A sample weighing 127mg was removed. Transferring the dishes to circulating airIn the dryer. The storage conditions were atmospheric pressure and 25 ℃. Will be similar to the wetting penetration gas flow (N) in example 2₂) Continuously introduced into the dryer. Using this technique, a relative humidity of 70% in the dryer can be ensured during storage. After about 48 hours the first sample (239 mg by mass) was taken, which represents the dihydrate form of the potassium salt (Ia) (XRPD analysis, thermogravimetric analysis). After a further 24 hours, a second sample (220 mg by mass) was taken. A sample of this stored material also represents the dihydrate form (XRPD analysis, thermogravimetric analysis). The mass remaining in the dish was 1.168g of the crystalline dihydrate form of the potassium salt (Ia).

Example 5:preparation of the anhydrous form of the crystalline potassium salt (Ia) from the monohydrate form by suspension in a suitable solvent

The monohydrate form of the potassium salt (Ia) (12.42g) was transferred to a flask. The monohydrate form of the substance was determined by XRPD-analysis and thermogravimetric analysis. 1, 4-dioxane solvent (125mL) was added to the flask, which was then blocked with an elastomeric membrane. The suspension was stirred with a magnetic stirrer and heated to 95 ℃ for 6 hours, after which the suspension was cooled to near room temperature and the crystals were separated from the solution with a suction filter. The wet cake was washed with 50mL of solvent (1, 4-dioxane, room temperature) and then dried in a vacuum drying chamber at 100mbar and 40 ℃. After a drying time of 44 hours 10.851g of the crystalline anhydrous form of the potassium salt (Ia) were produced (XRPD-analysis, thermogravimetric analysis, residual solvent).

While the invention has been described with reference to specific embodiments thereof, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition, process step or steps, to the objective, spirit and scope of the present invention. All such modifications are intended to be included within the scope of the appended claims.

Claims (15)

1. A crystalline potassium salt of 2- ((2S,3R,4E,6E,10E,12S) -13- (4-fluorophenoxy) -2,3, 12-trihydroxytridec-4, 6, 10-trien-8-ynyloxy) acetic acid,

   it comprises the following steps:

   an anhydrate form having an X-ray powder diffraction pattern with the following characteristic peaks:and

   the X-ray powder diffraction pattern has the monohydrate form with the following characteristic peaks:andor

   The dihydrate form of the X-ray powder diffraction pattern has the following characteristic peaks:
2. 2. a process for the synthesis of the crystalline potassium salt of claim 1

   The method comprises the following steps:

   1) mixing together a potassium base in a suitable solvent and a free acid corresponding to the potassium salt of formula (Ia) in a suitable solvent;

   2) optionally cooling the resulting suspension;

   3) separating the resulting crystals from the resulting suspension;

   4) optionally washing the isolated crystals with a suitable solvent; and

   5) the separated crystals were dried to obtain a crystalline potassium salt (Ia).
3. 3. A process as claimed in claim 2 wherein the suitable solvent for the potash comprises an organic solvent and water, and wherein the suitable solvent for the free acid comprises an organic solvent and water, and wherein the suitable solvent for washing the isolated crystals comprises an organic solvent and water.
4. 4. The method of claim 2 or 3, wherein the crystalline potassium salt is in the form of a monohydrate, a dihydrate, a dehydrated hydrate, a mixture of hydrates, or a mixture of hydrates and dehydrated hydrates.
5. 5. A process for synthesizing the crystalline anhydrate of the potassium salt according to claim 1

   The method comprises the following steps:

   1) digesting a hydrate or mixture of hydrates of the potassium salt of formula (Ia) in a suitable solvent;

   2) optionally heating the resulting suspension;

   3) separating the resulting crystals;

   4) optionally washing the isolated crystals with a suitable solvent; and

   5) the separated crystals were dried to obtain a crystalline anhydrate form of the potassium salt (Ia).
6. 6. A process as claimed in claim 5, wherein the suitable solvent for the hydrate or mixture of hydrates of the potassium salt comprises an organic solvent, and wherein the suitable solvent for washing the isolated crystals comprises an organic solvent.
7. 7. A pharmaceutical composition comprising one or more pharmaceutically acceptable excipients and a therapeutically effective amount of the crystalline potassium salt of claim 1.
8. 8. Use of a crystalline potassium salt of claim 1 in the manufacture of a medicament for treating a mammal having a disease state characterized by inflammation.
9. 9. The use of claim 8, wherein the mammal is a human.
10. 10. The use according to claim 8 or 9, wherein the disease state is an inflammatory disease or an autoimmune disease.
11. 11. The use of claim 10, wherein the inflammatory or autoimmune disease is selected from the group consisting of: allergic reactions, allergic contact dermatitis, allergic rhinitis, chemical and non-specific irritant contact dermatitis, urticaria, atopic dermatitis, psoriasis, fistulas associated with crohn's disease, pouchitis, infectious or endotoxic shock, hemorrhagic shock, shock-like syndrome, capillary leak syndrome induced by immunotherapy of cancer, acute respiratory distress syndrome, traumatic shock, immune-and pathogen-induced pneumonia, immune complex-mediated lung injury, chronic obstructive pulmonary disease, inflammatory bowel diseases including ulcerative colitis, crohn's disease and post-operative trauma, gastrointestinal ulcers, diseases associated with ischemia-reperfusion injury including acute myocardial ischemia and infarction, acute renal failure, ischemic bowel disease and acute blood loss or ischemic stroke, immune-complex-mediated glomerulonephritis, autoimmune diseases including insulin-dependent diabetes, multiple sclerosis, rheumatoid arthritis, osteoarthritis and systemic lupus erythematosus, acute and chronic organ transplant rejection, transplant atherosclerosis and fibrosis, cardiovascular diseases including hypertension, atherosclerosis, aneurysms, severe lower limb ischemia, peripheral arterial occlusive disease and raynaud's syndrome, diabetic complications including diabetic nephropathy, neuropathy and retinopathy, diseases of the eye including macular degeneration and glaucoma, neurodegenerative diseases including delayed neurodegeneration in stroke, alzheimer's disease, parkinson's disease, encephalitis and HIV dementia, inflammatory and neuropathic pain including arthritic pain, periodontal disease including gingivitis, ear infections, migraine, benign prostatic hyperplasia including but not limited to leukemia and lymphoma, Prostate cancer, breast cancer, lung cancer, malignant melanoma, renal cancer, head and neck tumors, and colorectal cancer.
12. 12. The use of claim 10, wherein the inflammatory or autoimmune disease is an inflammatory bowel disease selected from the group consisting of crohn's disease, pouchitis, ulcerative colitis, and gastrointestinal ulcers.
13. 13. The use according to claim 10, wherein the inflammatory or autoimmune disease is crohn's disease.
14. 14. The use according to claim 8 or 9, wherein the disease state is a pulmonary or respiratory inflammatory disease.
15. 15. The use according to claim 14, wherein the inflammatory disease of the lung or respiratory tract is selected from the group consisting of: asthma, chronic bronchitis, bronchiolitis obliterans including bronchiolitis with accompanying pneumonia, allergic inflammation of the respiratory tract including rhinitis and sinusitis, eosinophilic granuloma, pneumonia, pulmonary fibrosis, pulmonary manifestations of connective tissue disease, acute or chronic lung injury, chronic obstructive pulmonary disease, adult respiratory distress syndrome, and other noninfectious inflammatory diseases of the lungs characterized by eosinophilic infiltration.