Salts of Obeticholic Acid, Processes for their Preparation and their Intermediates

Abstract

Methods for preparing compounds of obeticholic acid and salts thereof, such as calcium salts thereof, are provided herein. Similarly, amorphous obeticholic acid hemicalcium is provided herein. Novel intermediates for use in the synthesis of obeticholic acid and salts thereof, including obeticholic acid HCl and mesylate salts for use in the synthesis of obeticholic acid and amorphous obeticholic acid hemicalcium, are also provided. The present disclosure provides amorphous calcium salts of obeticholic acid, such as amorphous obeticholic acid hemicalcium.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit and priority of U.S. Provisional Application No. 63 / 358,363, filed on July 5, 2023, which is hereby incorporated by reference in its entirety.

Background Art

[0002] Introduction Prospective epidemiological studies have shown a strong association between low - density lipoprotein - cholesterol (LDL - C) levels and the risk of cardiovascular disease (CVD). The application of statin therapy to lower these atherogenic LDL - C levels has led to a significant reduction in CVD - related morbidity and mortality: for every 1 mmol / L reduction in LDL - C, there is an estimated 22% reduction in CVD events and a 10% reduction in all - cause mortality. Despite such remarkable benefits, a large remaining disease burden persists, which has a major impact on both individual patients and the global healthcare costs. There is a need for new therapies to further reduce such remaining CVD risk in patients.

[0003] One way to lower LDL - C and increase high - density lipoprotein cholesterol (HDL - C) levels is to inhibit cholesterol ester transfer protein (CETP). CETP is a plasma protein mainly secreted by the liver and adipose tissue. CETP mediates the transfer of cholesteryl esters from HDL to apolipoprotein B (apoB) - containing particles (mainly LDL and very - low - density lipoprotein VLDL) during the exchange of triglycerides, thereby reducing the cholesterol content in HDL and preferentially increasing the cholesterol content in VLDL. Therefore, the hypothesis has been put forward that CETP inhibition retains cholesteryl esters in HDL - C and reduces the cholesterol content of the atherogenic apoB fraction.

[0004] Clinical trials have shown that obicetrapib, also known as ((2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester) or a pharmaceutically acceptable salt thereof, is a potent CETP inhibitor. The structure of obicetrapib is shown in the following formula (I): [Chemical formula] as described in

[0005] The preparation of obicetrapib is disclosed, for example, in U.S. Patent No. 7,872,126. Example 177 teaches the formation of obicetrapib as the free acid, as can be seen from formula (I). Example 178 teaches the formation of the sodium salt of obicetrapib by exchanging the acidic proton of the free acid moiety of obicetrapib with a sodium atom. In Example 179, the calcium salt of obicetrapib is taught. Since calcium is an alkaline earth metal, when ionized, it has a +2 charge. Therefore, the neutral salt will have two obicetrapib anions for each calcium cation (each being in a state where the carboxylic acid group of obicetrapib has no proton). The resulting salt in Example 179 is a hemicalcium salt in that it contains only half the number of calcium atoms as the number of obicetrapib anions in the neutral amorphous obicetrapib calcium salt molecule.

[0006] The molecular formula of amorphous obicetrapib hemicalcium is (C 32 H 30It is (Ca(N4O5F9)2). When discussing salts of obicetrapib, such as calcium salts, especially hemicalcium salts, it is understood that obicetrapib has lost one proton to form such salts. Thus, the term amorphous obicetrapib hemicalcium means that the salt moiety of each obicetrapib moiety is not of formula (I) (i.e., obicetrapib), but rather is formula (I) minus a proton. Example 179 explicitly teaches that the resulting hemicalcium salt of obicetrapib is crystalline. However, this crystalline form has undesirable properties such as poor physical stability.

[0007] Compared to other known CETP inhibitors, relatively low doses of obicetrapib are required to achieve nearly complete CETP inhibition. Typically, repeating a low once-daily dose of 2.5 mg of the obicetrapib compound has been shown to already be sufficient to achieve nearly complete CETP inhibition. These are substantially lower doses than those that must be used in the case of other CETP inhibitors. Furthermore, clinical trials have also shown that obicetrapib is well tolerated and does not cause serious side effects.

[0008] Methods for the manufacture of obicetrapib have been described (see, for example, WO2005 / 095409A2 and U.S. Pat. Nos. 7,872,126 and 8,158,640, Examples 1 and 177 - 180; WO2007 / 116922A1 and U.S. Pat. No. 8,084,611; and WO2016 / 024858 and U.S. Pat. No. 10,112,904), but the prior art produces disadvantageous solid forms. The entire contents of these references are incorporated herein by reference. Furthermore, the methods already described have relatively low yields and are not particularly suitable for implementation on an industrial scale. Accordingly, there is a need for alternative methods of manufacturing obicetrapib and pharmaceutically acceptable salts thereof with improved solid forms, as well as improved yields, purity, and stability.

Prior Art Documents

Patent Documents

[0009]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Patent Document 6

Patent Document 7

Summary of the Invention

Means for Solving the Problems

[0010] Summary of the Invention The present disclosure provides, for example, amorphous calcium salts of obeticholic acid such as amorphous obeticholic acid hemicalcium.

[0011] The present disclosure further provides HCl obeticholic acid containing a crystalline obeticholic acid HCl compound, and other compounds useful as intermediates in the production of obeticholic acid, such as the compound of formula (VI) and Compound 1D including crystalline Compound 1D.

[0012] The present disclosure provides methods for the production of (i) obeticholic acid including intermediates such as crystalline obeticholic acid HCl compound, (ii) other compounds useful as intermediates in the production of obeticholic acid, such as the compound of formula (VI) and Compound 1D including crystalline Compound 1D, and (iii) amorphous calcium salts of obeticholic acid such as amorphous obeticholic acid hemicalcium.

[0013] The present disclosure also provides a pharmaceutical composition comprising an amorphous obeticholic acid calcium salt, such as amorphous obeticholic acid hemicalcium, and one or more pharmaceutically acceptable carriers.

[0014] The present disclosure further provides a method of treating a patient suffering from, or having an increased risk of developing, a cardiovascular disease, the method comprising administering to such a patient an amorphous obeticholic acid calcium salt, such as amorphous obeticholic acid hemicalcium.

[0015] These and other features, aspects and advantages of the present invention will be better understood with reference to the following description and the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

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Mode for Carrying Out the Invention

[0044] Detailed Description of the Invention In many embodiments of the present disclosure, amorphous obeticholic acid calcium salts are provided. In particular, amorphous obeticholic acid hemicalcium is provided. The present disclosure further targets various methods for making amorphous obeticholic acid calcium, in particular, amorphous obeticholic acid hemicalcium.

[0045] The amorphous obeticholic acid calcium of the present disclosure is distinguishable from the crystalline obeticholic acid calcium disclosed in U.S. Patent No. 7,872,126. The general technique used to distinguish between a crystalline substance and an amorphous substance is X-ray powder diffraction. However, this technique has limitations, especially when the crystalline substance is irregular. In the case of amorphous obeticholic acid calcium, the X-ray powder diffraction patterns of two different lots of amorphous obeticholic acid calcium are presented in FIGS. 1 and 2. These patterns have the familiar "halo" type of features associated with amorphous substances. The X-ray powder diffraction pattern from FIG. 2 has peaks at approximately 3.4° (2θ), approximately 7.0° (2θ), and approximately 9.2° (2θ). Similarly, another sample in FIG. 3 also has X-ray powder diffraction peaks at approximately 3.4° (2θ), approximately 7.0° (2θ), and approximately 9.2° (2θ). Any of the X-ray powder diffraction patterns of FIG. 1 or FIG. 2 or FIG. 3 can be used to characterize amorphous obeticholic acid calcium, provided that occasionally, sharp and even larger angle peaks are present, such as those at approximately 31.7° (2θ) (as in FIG. 2), and when present, those peaks are due to sodium chloride. In FIG. 3, for another sample of amorphous obeticholic acid calcium, peaks at approximately 3.4° (2θ), approximately 7.0° (2θ), and approximately 9.2° (2θ) were identified. The peak at approximately 5.6° (2θ) in FIG. 3 was determined to be due to the Kapton foil used in the measurement setup described in Example 20. The X-ray powder pattern of the crystalline obeticholic acid calcium prepared in Example 16 is shown in FIG. 6. It also exhibits halo-like behavior, which can be an indicator of irregularity in the case of a crystalline compound.

[0046] Examples 18, 19, 20, 21 and 22 describe various X-ray powder diffraction procedures. The procedure of Example 18 was generally used to collect the data illustrated in FIGS. 1, 6, 7 and 8. Example 19 was generally used for FIG. 2. Example 20 was generally used for FIG. 3. Example 21 was generally used for FIGS. 18, 19 and 20 (FIG. 20 is for Compound 1D rather than crystalline obeticholic acid HCl). Example 22 was generally used for FIGS. 22-25 and FIG. 28.

[0047] The use of the term "amorphous" in "amorphous obeticholic acid hemicalcium" does not mean that this substance never has regularity. As indicated by the presence of peaks in the X-ray powder diffraction pattern, there is still a certain degree of regularity in the sample. Thus, as used herein, the term "amorphous" in "amorphous obeticholic acid hemicalcium" does not mean that the X-ray powder diffraction pattern must purely contain an amorphous halo (however, it may have halo-like features). Rather, the above term means that irregularities exist, but the amorphous phase is distinguishable from the crystalline phase discussed below.

[0048] Another technique that can be used to distinguish between a crystalline substance and an amorphous substance is polarized light microscopy ("PLM"). In PLM, the substance is observed through polarized light, and by observing the substance through crossed polarizers, it is possible to distinguish whether the substance is anisotropic (e.g., crystalline) or isotropic (e.g., amorphous compound). An anisotropic substance exhibits birefringence by showing a color change through the polarizer when exposed to the polarized light that has passed through the polarizer. On the other hand, an isotropic substance does not exhibit birefringence and does not show a color change even when exposed to polarized light.

[0049] In Figure 9, amorphous obcetrapib hemicalcium was analyzed by polarized light optical microscopy as described in Example 17. As shown in Figure 9, this substance does not exhibit birefringence under examination, indicating that this substance is amorphous. By comparison, Figure 10 is a polarized light micrograph of crystalline obcetrapib hemicalcium prepared according to Example 16. In particular, the particles shown in Figure 10 (which are black and white) exhibit a considerably brighter contrast. In the corresponding color version, the figure is multicolored. Thus, Figure 10 indicates crystallinity. Furthermore, the crystals in Figure 10 are larger than the particles presented in the polarized light micrograph of the amorphous obcetrapib hemicalcium in Figure 9. Therefore, amorphous obcetrapib hemicalcium can be characterized using the absence of PLM and / or birefringence.

[0050] Other techniques can be further used to distinguish amorphous obcetrapib hemicalcium from crystalline obcetrapib hemicalcium, and thus amorphous obcetrapib hemicalcium can be characterized using this other technique. One such technique is modulated differential scanning calorimetry, also referred to as "mDSC". The difference in the heat required to raise the temperature of the sample, compared to a reference, is measured as a function of temperature and can be measured using modulated differential scanning calorimetry (mDSC). In an mDSC thermogram, the glass transition temperature, which can be used to characterize amorphous substances, can also be measured. In Figure 12, with respect to this figure, the procedure is described in Example 25, and the mDSC thermogram of amorphous obcetrapib hemicalcium was measured using an open sample holder, which allows volatile gases to be expelled during the measurement. In Figure 12, the opening was made by punching a hole in the lid of the pan to create a pinhole. In the case of this sample, a glass transition temperature of approximately 110 °C was recorded.

[0051] Regarding thermal measurements, the term "about" generally refers to a variation of plus or minus 1°C. In comparison, crystalline obcetrapib hemicalcium has a higher glass transition temperature under the same conditions, and the three measurements in Figure 14 indicate a range between about 118°C and about 125.5°C. In some embodiments, the glass transition temperature of amorphous obcetrapib hemicalcium is between about 109°C and 112°C when measured with pinholes provided. In one of the samples, in Example 26, it is found that the glass transition temperature of amorphous obcetrapib hemicalcium is about 111°C (111.32°C at the midpoint), as shown in Figure 13. This start was measured at about 102°C (101.62°C) and the end point at about 118°C (117.58°C).

[0052] The glass transition temperature of amorphous obcetrapib hemicalcium can also be measured using mDSC with a hermetic pan. The type of sample preparation may affect the measured glass transition temperature. In such cases, the glass transition temperature decreases to less than about 100°C, particularly to a temperature between about 70°C and about 92°C, depending on the humidity.

[0053] Other thermal techniques such as thermogravimetric analysis (TGA) can also be used to analyze and characterize amorphous obcetrapib hemicalcium. Figure 11 is a thermogram of the thermogravimetric analysis of amorphous obcetrapib hemicalcium, showing a weight loss of less than 1% when heated to about 200°C. Such weight loss can be in the range of about 0.8% to about 0.95%, including, for example, between about 0.84% and about 0.92%. In Figure 11, the weight loss was determined to be about 0.85%. Such a specific substance was found to have a moisture content of about 1.5%. In some embodiments, the moisture content may be higher, including a range of water from about 0% to about 5%, up to about 4% by weight, up to about 3% by weight, and between about 0.5% and 1.5% by weight.

[0054] Solid state 1313C-NMR spectroscopy is another technique that can be used to characterize amorphous substances. Figure 15 shows the solid state of both crystalline obeticholic acid hemicalcium and amorphous obeticholic acid hemicalcium. 13 13C-NMR spectra are shown, and Figures 16 and 17 show crystalline obeticholic acid hemicalcium and amorphous obeticholic acid hemicalcium, respectively. There are at least two differences in the spectra. The crystalline phase has a peak at about 22.1 ppm, which is not present in the amorphous phase. Furthermore, the peak at about 29.5 ppm in the crystalline phase is prominent, while in the amorphous phase it is not. Thus, the absence of a solid state 13 13C-NMR peak at about 22.1 ppm and / or the absence of a prominent peak at about 29.5 ppm can be used to characterize amorphous obeticholic acid hemicalcium. Furthermore, the solid state 13 13C-NMR spectrum that is substantially the same as the solid state 13 13C-NMR spectrum can be used to characterize amorphous obeticholic acid hemicalcium. The absence of a peak in this situation does not mean, for example, that the intensity at 22.1 ppm or 29.5 ppm necessarily does not exist. Rather, the intensity is not prominent as in the 13 13C-NMR spectrum of crystalline obeticholic acid hemicalcium.

[0055] The properties of crystalline substances are usually also different from those of amorphous substances. Thermodynamically, crystalline substances are physically more stable than amorphous substances. Thus, there is a thermodynamic driving force to convert amorphous compounds to crystalline compounds. Thus, if a physical transformation of the solid form is thought to exist, under accelerated stress conditions, the solid form is generally expected to convert from amorphous to crystalline. However, in the case of obeticholic acid hemicalcium, the opposite is true.

[0056] Figure 6 is a plot of the measured x-ray powder diffraction pattern of crystalline obcetrapib hemicalcium, and Figure 7 is a plot of the x-ray powder diffraction pattern of crystalline obcetrapib hemicalcium measured under stress conditions. In Figure 7, there are four diffraction patterns shown based on the stability study described in Example 27. Pattern 1 is the x-ray powder diffraction pattern of a sample of amorphous obcetrapib hemicalcium. Pattern 2 is the x-ray powder diffraction pattern of a sample of crystalline obcetrapib hemicalcium. In Pattern 3, a sample of crystalline obcetrapib hemicalcium was exposed to 70 °C at 75% relative humidity for 1 day. As can be seen from Pattern 3, the x-ray powder diffraction pattern indicates that the crystallinity was almost completely lost on that day. After 7 days under the same conditions, the results are still in the same state as those observed in Pattern 4. A similar experiment was conducted on the amorphous obcetrapib hemicalcium shown in Figure 8. Pattern 1 was measured before the sample reached a stable state. Even when the substance was exposed to the same conditions of 70 °C and 75% relative humidity, crystallization was not induced, and the substance remained amorphous after 7 days (Pattern 2) and 14 days (Pattern 3). Therefore, these experiments suggest that, contrary to what was expected, the amorphous form of obcetrapib hemicalcium is more physically stable than the crystalline obcetrapib hemicalcium.

[0057] In some embodiments of the present disclosure, stable amorphous obcetrapib hemicalcium is provided herein. In these embodiments, the amorphous obcetrapib hemicalcium is physically more stable than the crystalline obcetrapib hemicalcium under typical pharmaceutical use and processing conditions.

[0058] While not desiring to be bound by theory, at least with respect to pharmacologically relevant processes and thermodynamic conditions under which a more stable crystalline phase exists, in this case, a kinetics such that the amorphous phase is kinetically stabilized is contemplated. The result of such a stability profile is that amorphous obeticholic acid hemicalcium is more suitable for pharmaceutical development and use than the corresponding crystalline phase. Despite being more physically adaptable, amorphous obeticholic acid hemicalcium has a higher solubility than highly insoluble crystalline obeticholic acid hemicalcium.

[0059] Solubility is, inter alia, a problem in the case of obeticholic acid. At 20° C., for example, the solubility of obeticholic acid in water was measured to be substantially less than 0.1 mg / mL. It would be desirable to have a solid form of obeticholic acid that delivers a large amount of obeticholic acid.

[0060] Solubility is a thermodynamic quantity of a substance, but the kinetic solubility of a substance can be measured without necessarily reaching thermodynamic equilibrium. Such measurements result in a solubility under metastable conditions and, for example, provide information on the amount of substance undergoing dissolution as a function of time.

[0061] The amorphous form has a higher kinetic solubility and dissolution rate than the crystalline form (and, by extension, obeticholic acid itself). The determination of the kinetic solubility of both crystalline obeticholic acid hemicalcium and amorphous obeticholic acid hemicalcium was performed in a biorelevant medium at various pHs, namely, at a pH of about 5.0 (FeSSIF conditions) and about 6.5 (FaSSIF conditions) as described in Example 28.

[0062] Table 1 shows the measured solubilities of two different batches of amorphous obeticholic acid hemicalcium in FeSSIF medium at 37 °C over a 2-hour period. In both cases, the amorphous obeticholic acid hemicalcium had a higher concentration in solution than the corresponding crystalline material at all measured time points. The concentrations in Table 1 are those of obeticholic acid (i.e., the free acid).

Table 1

[0063] Table 2 shows a similar experiment at 37 °C but in FaSSIF medium at a pH of 6.5. As in Table 1, in both batches, the amorphous obeticholic acid hemicalcium had a higher concentration in solution than the corresponding crystalline material at all measured time points. The concentrations in Table 2 are those of obeticholic acid (i.e., the free acid).

Table 2

[0064] Amorphous obeticholic acid hemicalcium is also advantageous in that, unlike many amorphous organic compounds, it does not readily take up moisture. For example, when exposed to a relative humidity close to 90%, usually less than about 5% moisture uptake is measured. The lack of such hygroscopicity is advantageous as it does not require special handling or storage conditions. Other drawbacks commonly associated with the manufacture and use of amorphous substances are also absent. For example, amorphous substances are often difficult to make chemically pure. However, here, amorphous obeticholic acid hemicalcium with a chemical purity of 99.9% or higher can be routinely prepared.

[0065] In some embodiments of the present disclosure, substantially pure amorphous obeticholic acid hemicalcium is provided. In these and other embodiments, the chemical purity of the substantially pure amorphous obeticholic acid hemicalcium is 99.9% or higher.

[0066] In many aspects of the present disclosure, a method for preparing an amorphous calcium salt of obeticholic acid, such as amorphous obeticholic acid hemicalcium, comprising the steps of treating obeticholic acid with an acid to form a salt, solvate or composition, isolating the resulting salt, solvate or composition, and treating such salt, solvate or composition with a calcium source to produce an amorphous calcium salt of obeticholic acid, such as amorphous obeticholic acid hemicalcium, is provided. The resulting salt can then be isolated.

[0067] Examples of calcium sources include calcium salts such as calcium halide salts and soluble calcium salts. In many embodiments, the calcium source is calcium chloride.

[0068] The preparation of amorphous salts of obeticholic acid calcium, such as amorphous obeticholic acid hemicalcium, has been found to be carried out in the presence of intermediate salts, solvates or compositions (such compositions contain the corresponding acids used to make the salts). Treating obeticholic acid directly with a calcium base such as calcium hydroxide is not a feasible way to make amorphous salts of obeticholic acid calcium because of either low solubility, weak available base, or both. Rather, it has been found that the preparation of amorphous obeticholic acid hemicalcium is achievable by using an intermediate salt such as a sodium salt. However, even when using a sodium salt, it is preferred to utilize an additional salt or salt type exchange associated with the sodium salt of obeticholic acid (more so by using a composition or solvate rather than the actual salt) for purity and yield purposes. In particular, the use of salts, solvates or compositions enables the production of very pure amorphous calcium salts of obeticholic acid such as amorphous obeticholic acid hemicalcium.

[0069] Exemplary salts that may be prepared as intermediates include sulfonates (e.g., besylate, tosylate, napsylate, camsylate, esylate, edisylate or mesylate), sulfates (e.g., methyl sulfate), halogens (e.g., chloride ion, iodide ion or bromide ion), acetates, aspartates, benzoates, bicarbonates, bitartrates, carbonates, citrates, decanoates, fumarates, gluceptates, gluconates, glutamates, glycolates, hexanoates, hydroxynaphthoates, isethionates, lactates, lactobionates, malates, maleates, mandelates, mucates, nitrates, octanoates, oleates, pamoates, pantothenates, phosphates, polygalacturonates, propionates, salicylates, stearates, succinates, tartrates or theophyllinates. When the intermediate is a solvate or composition, the corresponding acid may be used or may be present. Further, in the case of a solvate, the intermediate may further contain a solvent such as an organic solvent or water, and in the latter case, the solvate becomes a hydrate. One such organic solvent is CPME (cyclopentyl methyl ether).

[0070] In some embodiments, the intermediate is a solvate of an acid. In these and other embodiments, the intermediate is a solvate of an acid and an organic solvent. In some particular embodiments, the intermediate is a solvate containing an acid and a solvent. In some of these embodiments, the acid is hydrochloric acid and the solvent is CPME.

[0071] In many aspects of the present disclosure, the present disclosure includes a method for preparing an amorphous obicetrapib calcium salt such as amorphous obicetrapib hemicalcium. The present disclosure further includes an amorphous obicetrapib calcium salt containing amorphous obicetrapib hemicalcium thus prepared. In one such preparation, an intermediate herein referred to as crystalline obicetrapib HCl is used in a method for preparing an amorphous obicetrapib calcium such as amorphous obicetrapib hemicalcium.

[0072] In many aspects of the present disclosure, amorphous obeticholic acid hemicalcium is prepared by chemical synthesis and has the formula (IH): [Chemical formula] The intermediate represented by is used. y varies such that the mass percentage of HCl varies from 0.01 wt% to 8 wt% and is considered to further include an organic solvent associated, such as by a solvate. In some embodiments, y varies from 0.002 to 1.5. In some embodiments, y varies from 0.3 to 1. In some embodiments, y varies from 0.4 to 0.6, including between 0.5 and 0.6. In some embodiments, the formula (IH) as a solvate is isolated in its crystalline form. In many embodiments, the solvent is CPME. Other solvents capable of forming solvates include toluene and heptane.

[0073] As used herein, obeticholic acid HCl prepared is usually crystalline. Further, the term crystalline obeticholic acid HCl can include CPME as a solvate when CPME is used in the preparation of crystalline obeticholic acid HCl. In formula (IH), the solvate is a solvate of an organic solvent, and in many embodiments, the solvent is CPME. In some embodiments, the present disclosure provides a composition comprising crystalline obeticholic acid HCl.

[0074] The formula (IH) is called obeticholic acid HCl and, when crystalline, is referred to as crystalline obeticholic acid HCl. One crystal structure of the solid form of crystalline obeticholic acid HCl has been elucidated and is consistent with crystalline obeticholic acid HCl of form B.

[0075] The crystal structure was elucidated according to Example 34, and single crystals were prepared according to Example 33. This structure is a complex multi-component crystal, showing six obicetrapib moieties, two of which are neutral and four of which are charged in the asymmetric unit. There are four protonated obicetrapib molecules, each protonated at the nitrogen on the pyrimidine ring of obicetrapib, and four chloride ions. Two chloride ions each appear to associate with two of the protonated nitrogens, evidencing salt formation, while the other two chloride ions coordinate to the carboxyl moieties. This structure contains heptane and cyclopentylmethyl ether (CPME) solvent molecules. There are void spaces of unknown occupancy, with sufficient room for solvent and / or HCl to enter, but no room for obicetrapib. Without being bound by theory, crystalline HCl obicetrapib is considered to be a mixed salt solvate. In the reaction to produce formula (IH), when CPME is used to supply HCl, the chloride ion content of formula (IH) is found to be in the range between about 2.5 wt% and 3.0 wt%, which is found to be below what would be expected for a neutral salt (i.e., about 4.8 wt%).

[0076] In many embodiments, when CPME is so used and upon crystallization, CPME is found in the substance. When CPME is used in the reaction to supply dry HCl and thus is found in the crystallized substance, the resulting crystalline Form (IH) substance is referred to as crystalline obeticholic acid HCl, and their X-ray powder diffraction patterns are shown in Figure 18. The advantage of using crystalline obeticholic acid HCl as an intermediate is that the resulting amorphous obeticholic acid hemicalcium typically has a chemical purity of 99.9% or higher. Chemical purity is a quantitative value representing whether other chemical entities other than the compound being measured are present. For example, amorphous obeticholic acid hemicalcium with a chemical purity of 99.9% means that less than 0.1% of the compounds in a sample of amorphous obeticholic acid hemicalcium are other entities. Physical purity refers to the amount of other solid forms of the same compound present, which, in the case of amorphous obeticholic acid hemicalcium, the other solid form is crystalline obeticholic acid hemicalcium. The disclosure herein provides physically pure amorphous obeticholic acid hemicalcium, which means it contains no or substantially no crystalline obeticholic acid hemicalcium. Unless otherwise specified herein, the purity measurements presented herein are measurements of chemical purity.

[0077] Obeticholic acid HCl, as used herein, is not limited to crystalline obeticholic acid HCl. In fact, crystalline obeticholic acid HCl may become amorphous upon desolvation.

[0078] Upon stress, crystalline obcetrapib HCl loses its crystallinity. In Figure 18, Pattern 2 reflects the mildly dried crystalline obcetrapib HCl, by which the surface solvent is removed and it can be seen that this compound is crystalline. By comparison, the sample whose X-ray powder diffraction is measured in Pattern 1 was subjected to a more intensive drying treatment at 55 °C for 48 hours at a pressure of 2 mbar. Clearly, such drying probably changed this substance from crystalline to amorphous due to the loss of HCl and the desolvation of CPME. For example, 1 Using H-NMR spectroscopy, it was shown that in the upper pattern, CPME is present, while in the lower amorphous pattern, it is substantially absent. Thus, the amorphous pattern represents HCl obcetrapib that is not crystalline obcetrapib. It may be obcetrapib, but is thought to have HCl associated with obcetrapib as a solvate, and thus is HCl obcetrapib, but has a lower chloride content than is normally observed in the range observed in the case of crystalline obcetrapib HCl. In some embodiments, the chloride content is less than 0.1 wt%, such as between about 0.01 wt% and 0.1 wt%.

[0079] Crystalline obcetrapib HCl can be characterized by an X-ray powder diffraction pattern that includes a peak at about 9.8° (2θ). In some embodiments, crystalline obcetrapib HCl can be characterized by an X-ray powder diffraction pattern that includes one or more peaks at about 8.1° (2θ), about 9.8° (2θ), about 13.8° (2θ), about 16.7° (2θ), or about 19.5° (2θ). Table 3 presents exemplary peaks that may be present in crystalline obcetrapib HCl. In some embodiments, crystalline obcetrapib HCl can be characterized by an X-ray powder diffraction pattern that is substantially the same as the X-ray powder diffraction pattern in Figure 19.

[0080] The substance analyzed in FIG. 19 was measured such that no peak was measured between about 4.3° (2θ) and about 4.7° (2θ). In fact, in each of the forms A, B, C, and D of crystalline obcetrapib hydrochloride, a peak is present and is located between about 4.3° (2θ) and about 4.7° (2θ). Thus, crystalline obcetrapib hydrochloride can be characterized by an x-ray powder diffraction pattern that includes a peak between about 4.3° (2θ) and about 4.7° (2θ). [Table 3]

[0081] Multiple forms of crystalline obcetrapib hydrochloride are disclosed herein. In some embodiments, crystalline obcetrapib hydrochloride of form A is provided. The preparation of crystalline obcetrapib hydrochloride of form A is described in Example 29. The x-ray powder diffraction pattern of crystalline obcetrapib hydrochloride of form A is presented in FIG. 22. Table 4 presents exemplary peaks that may be present in crystalline obcetrapib hydrochloride of form A. [Table 4-1] [Table 4-2]

[0082] In some embodiments, crystalline obcetrapib hydrochloride of form A can be characterized by an x-ray powder diffraction pattern that includes a peak at about 8.6° (2θ), the presence of two peaks between about 9.7° (2θ) and about 10.4° (2θ), and the presence of two peaks at about 8.6° (2θ) and about 9.0° (2θ). In some embodiments, crystalline obcetrapib hydrochloride of form A has an x-ray powder diffraction pattern that is substantially the same as the x-ray powder diffraction pattern of FIG. 22.

[0083] In some embodiments, crystalline obcetrapib hydrochloride of Form B is provided. The preparation of crystalline obcetrapib hydrochloride of Form B is described in Example 30. The X-ray powder diffraction pattern of crystalline obcetrapib hydrochloride of Form B is presented in Figure 23. Table 5 presents exemplary peaks that may be present in crystalline obcetrapib hydrochloride of Form B. [Table 5]

[0084] In some embodiments, crystalline obcetrapib hydrochloride of Form B can be characterized by an X-ray powder diffraction pattern that includes peaks at approximately 6.5° (2θ), approximately 8.8° (2θ), and approximately 11.0° (2θ). In some embodiments, crystalline obcetrapib hydrochloride of Form B has an X-ray powder diffraction pattern that is substantially the same as the X-ray powder diffraction pattern of Figure 23.

[0085] In some embodiments, crystalline obcetrapib hydrochloride of Form C is provided. The preparation of crystalline obcetrapib hydrochloride of Form C is described in Example 31. The X-ray powder diffraction pattern of crystalline obcetrapib hydrochloride of Form C is presented in Figure 24. Table 6 presents exemplary peaks that may be present in crystalline obcetrapib hydrochloride of Form C. [Table 6]

[0086] In some embodiments, crystalline obcetrapib hydrochloride of Form C has an X-ray powder diffraction pattern that is substantially the same as the X-ray powder diffraction pattern of Figure 24.

[0087] In some embodiments, crystalline obcetrapib hydrochloride of Form D is provided. The preparation of crystalline obcetrapib hydrochloride of Form D is described in Example 32. The X-ray powder diffraction pattern of crystalline obcetrapib hydrochloride of Form D is presented in Figure 25. Table 7 presents exemplary peaks that may be present in crystalline obcetrapib hydrochloride of Form D. [Table 7]

[0088] In some embodiments, the crystalline obcetrapib hydrochloride of Form D has an x-ray powder diffraction pattern that is substantially the same as the x-ray powder diffraction pattern of FIG. 25.

[0089] Single crystal x-ray diffraction is another technique that can be used to elucidate the structure of crystalline materials. The solution of the single crystal structure showing the asymmetric unit of the crystals of crystalline obcetrapib hydrochloride prepared according to Example 33 is presented in FIG. 26. The unit cell parameters are found in Table 8 below.

Table 8

[0090] Although irregular, this solution results in a pattern similar to that of the crystalline obcetrapib hydrochloride of Form B, despite the use of a solvent system that can result in the crystalline obcetrapib hydrochloride of Form A in the preparation. This solution shows four protonated obcetrapib moieties (each protonated at the N3 nitrogen on the pyrimidine ring), and each of the four chloride anions is N-H at N3 of the pyrimidine ring in the lattice unit + is hydrogen bonded to the unit. This structure further contains two neutral obcetrapib molecules, and solvent molecules of heptane and CPME. One CPME molecule and one heptane molecule are thought to be present in the unit cell. There is at least one additional heptane molecule, and additional void space that can contain up to four additional heptane molecules as a possibility.

[0091] This crystal structure suggests a complex variable structure associated with crystalline obcetrapib hydrochloride. It is a solvate considering the presence of CPME and heptane.

[0092] Using single crystal data, the powder pattern that can be observed in Figure 27 was calculated. The superposition of the calculated pattern, crystalline obcetrapib hydrochloride of Form A and x-ray powder diffraction of crystalline obcetrapib hydrochloride of Form B is shown in Figure 28. Pattern 1 is associated with crystalline obcetrapib hydrochloride of Form A, Pattern 2 is associated with the calculated pattern, and Pattern 3 is associated with crystalline obcetrapib hydrochloride of Form B. Pattern 2 appears to be similar to Pattern 3.

[0093] Another intermediate used in the preparation of obcetrapib has the formula (VI)

Chemical formula

[0094] In one embodiment, the compound of formula (VI) has n = 1, Y 1 is t-butyl, and the compound is 1D:

Chemical formula

[0095] Of compound 1D 1The 1H-NMR spectrum (in solution) can be seen in Figure 21. Crystalline Compound 1D can be characterized by an X-ray powder diffraction pattern that includes one or more peaks at approximately 5.2° (2θ) or approximately 9.1° (2θ). In some embodiments, Crystalline Compound 1D can be characterized by an X-ray powder diffraction pattern that includes one or more peaks at approximately 5.2° (2θ), approximately 9.1° (2θ), approximately 15.9° (2θ), approximately 16.5° (2θ), approximately 17.2° (2θ), approximately 18.6° (2θ), and approximately 19.2° (2θ). Table 9 presents exemplary peaks that may be present in Crystalline Compound 1D (the peak at approximately 5.2° (2θ) was not measured due to the limitations of the instrument in reflection mode). In some embodiments, Crystalline Compound 1D can be characterized by an X-ray powder diffraction pattern that is substantially the same as that in Figure 20.

Table 9-1

Table 9-2

[0096] For example, crystalline compounds such as crystalline compound 1D and crystalline obeticholic acid HCl can be characterized by X-ray powder diffraction. The X-ray powder diffraction pattern is an x-y graph having °2θ (diffraction angle) on the x-axis and intensity on the y-axis. Peaks are usually represented and referenced by their position on the x-axis rather than by the intensity of the peaks on the y-axis. This is because peak intensity can be particularly sensitive to the orientation of the sample (see Pharmaceutical Analysis, Lee & Web, pp. 255-257 (2003)). Thus, intensity is not usually used for the characterization of solid forms. Data from X-ray powder diffraction can be used in multiple ways to characterize the crystal form. For example, the entire output value of the X-ray powder diffraction pattern from a diffractometer can be used to characterize the crystalline obeticholic acid HCl compound or crystalline compound 1D. However, smaller subsets of such data can also be suitable, and usually can be suitable, for characterizing such compounds. For example, collections of one or more peaks derived from such patterns can be used to characterize these compounds in such a way. When the phrase "one or more peaks" from a list of peaks from an X-ray powder diffraction pattern is presented, generally what is meant is that any combination of the listed peaks may be used for the characterization. Further, the presence of other peaks in the X-ray powder diffraction pattern generally does not negate such a characterization, nor does it limit such a characterization in any other way.

[0097] In addition to the variation in peak intensity, there may also be variation in the position of the peak on the x-axis. However, this variation can usually be taken into account when reporting the position of the peak for characterization purposes. Such variation in the position of the peak along the x-axis can be due to several causes (e.g., sample preparation, particle size, moisture content, solvent content, instrument parameters, data analysis software, and sample orientation). For example, samples of the same crystalline substance prepared under different conditions may give slightly different diffractograms, and different x-ray instruments can be operated using different parameters, which can result in slightly different diffraction patterns from the same crystalline solid. Due to such causes of variation, it is common to describe the x-ray diffraction peak by using the word "about" in front of the peak value (°2θ). For the purposes of the data reported herein, the fact that the value is generally ±0.2° (2θ) is always intended to be reported with such variation when disclosed herein, regardless of whether the word "about" is present. The variation can be higher in some cases depending on the instrument conditions, including how well the instrument is maintained.

[0098] In some embodiments, the crystalline compound 1D can be further characterized by having an x-ray powder diffraction pattern that is substantially the same as the x-ray powder pattern of FIG. 20.

[0099] In many aspects of the present disclosure, a method for preparing an amorphous calcium salt of obeticholic acid, such as amorphous obeticholic acid hemicalcium i. treating obeticholic acid with HCl to obtain crystalline obeticholic acid HCl, ii. isolating the crystalline obeticholic acid HCl, iii. preparing an amorphous calcium salt of obeticholic acid, such as amorphous obeticholic acid hemicalcium, from the crystalline obeticholic acid HCl isolated in step (ii), and iv. the step of isolating an amorphous calcium salt of obetrapib, such as amorphous obetrapib hemicalcium A method is provided that includes this step.

[0100] In another aspect of the present disclosure, a method for preparing obetrapib, comprising: (a) preparing a compound of formula (IV) by coupling a compound of formula (II) or a salt thereof with a compound of formula (III);

Chemical formula

Chemical formula

Chemical formula

[0101] ​The reactions in steps (a) to (d) of the subject method are carried out in a solvent, and the intermediate compounds of formulas (IV), (V) and (VIII) need not be isolated from their individual solvents if they are to be further processed to the final product. This means that by evaporating at least a part of the solvent used in step (x) and adding the solvent of step (x + 1) step by step, some kind of solvent exchange is carried out between reaction step (x) and reaction step (x + 1) so that the compound remains in solution during the solvent exchange. The intermediate compound of formula (VI) may be isolated from the solvent as a salt in solid form, and thus this salt can be washed to remove impurities. This isolation step ensures a downstream product of sufficient purity. The subject method need not include a purification step using chromatography such as column chromatography to achieve the chemical purity levels described herein. Method for preparing amorphous calcium salts (such as amorphous obeticholic acid hemicalcium) - Steps (i) to (ii) from embodiments (i) to (iv)

[0102] In some embodiments of the method for preparing an amorphous calcium salt of obeticholic acid such as amorphous obeticholic acid hemicalcium, the method includes step (i), i.e., treating obeticholic acid in an organic solvent with HCl to obtain crystalline obeticholic acid HCl.

[0103] In some embodiments, the crystalline obeticholic acid HCl has a purity of 98% or higher, such as 98.5% or higher, 99% or higher, 99.5% or higher, or even higher.

[0104] In some embodiments, the HCl in step (i) is present in a suitable solvent. Such a solvent may be an aqueous solvent or an organic solvent. In some embodiments, the organic solvent used in step (i) comprises a mixture of a solvent and an anti-solvent. In some embodiments, the solvent is selected from methanol, ethanol, isopropanol, acetic acid, acetonitrile, acetone, methyl isobutyl ketone, isopropyl acetate, tetrahydrofuran, methyl t-butyl ether, cyclopentyl methyl ether, N-methyl-2-pyrrolidone, dimethyl sulfoxide, dimethylformamide, 2-methyl-tetrahydrofuran, dichloromethane, 1,4-dioxane, 1,2-diflurobenzene, toluene, hexafluoroisopropanol, and water. In some embodiments, the anti-solvent is selected from n-heptane, n-hexane, n-pentane, and cyclohexane.

[0105] In some embodiments, the HCl has sufficient solubility in the anti-solvent such that the anti-solvent can be used as a suitable solvent. In some embodiments, the organic solvent used in step (i) comprises a mixture of cyclopentyl methyl ether and n-heptane. In some embodiments, the organic solvent used in step (i) further comprises toluene. In some embodiments, toluene is the major component of the organic solvent.

[0106] In some embodiments, step (i) includes preparing obicetrapib in a mixture of cyclopentyl methyl ether and n - heptane, raising the temperature to between 35 °C and 40 °C while stirring, adding dry HCl in cyclopentyl methyl ether and raising the temperature again to between 50 °C and 55 °C, and then adding additional n - heptane as an anti - solvent. At this point, a small amount of the reaction mixture can be extracted as needed, cooled to a temperature between 10 °C and 15 °C, and a slurry of crystals of crystalline obicetrapib HCl (referred to herein as the "seed crystal slurry") can be obtained in a mixture of cyclopentyl methyl ether and n - heptane. Optionally, all or part of the seed crystal slurry of crystalline obicetrapib HCl can then be added back to the reaction mixture. This species aids in nucleation but is not necessary. Next, the resulting reaction mixture is cooled to a temperature between 5 °C and 15 °C (such as 10 °C - 15 °C), and then crystalline obicetrapib HCl is crystallized from the system while stirring. In some embodiments, crystalline obicetrapib HCl is crystallized over 12 hours or longer, then filtered (e.g., through a filter dryer), washed one or more times as needed with a mixture of cyclopentyl methyl ether and n - heptane, etc., and dried. In some cases, the wet filter cake of crystalline obicetrapib HCl is dried step - wise under vacuum using temperatures such as 25 °C, 35 °C, 46 °C, and 54 °C, i.e., 25 °C - 30 °C, 30 °C - 40 °C, 40 °C - 50 °C, and then 50 °C - 55 °C.

[0107] In some embodiments, the method for preparing crystalline obicetrapib HCl includes the addition of seed crystals (e.g., as a seed crystal slurry). The seed crystals of the HCl compound can be formed as a slurry by extracting a small amount of the reaction mixture after the addition of dry HCl in cyclopentyl methyl ether and n - heptane as an anti - solvent, cooling to a temperature between 10 °C and 15 °C, and obtaining a slurry of crystals of crystalline obicetrapib HCl in cyclopentyl methyl ether and n - heptane, according to step (i) described above.

[0108] Thus, in one embodiment, step (i) comprises the steps of preparing crystalline obeticholic acid HCl in a mixture of cyclopentyl methyl ether and n-heptane, raising the temperature to between 35° C. and 45° C. while stirring, adding dry HCl in cyclopentyl methyl ether, and raising the temperature again to between 50° C. and 55° C., adding additional n-heptane as an anti-solvent, and adding seed crystals of the HCl compound (e.g., as a seed crystal slurry prepared as described herein) as needed, cooling to a temperature between 5° C. and 15° C. (such as between 10° C. and 15° C.), and then crystallizing crystalline obeticholic acid HCl from this system while stirring. In some embodiments, crystalline obeticholic acid HCl is crystallized over 12 hours or longer, filtered, washed one or more times as needed with a mixture of cyclopentyl methyl ether and n-heptane, etc., and drying follows. In some embodiments, crystalline obeticholic acid HCl is dried under vacuum. In some embodiments, crystalline obeticholic acid HCl is dried in a vacuum drying cabinet at a pressure of 25 mbar and a temperature of 55° C. for 10 hours or longer. In some embodiments, after the drying procedure, crystalline obeticholic acid HCl contains less than 0.1 wt % residual cyclopentyl methyl ether.

[0109] In some embodiments, step (i) involves preparing a solution of obicetrapib in cyclopentyl methyl ether at a concentration between 30% and 40% by weight, such as 33% to 37% by weight, based on the weight of the solution (wherein the first organic solvent (such as toluene) used in step (d) is less than 1% by weight and n-heptane is less than 1% by weight), adding n-heptane, raising the temperature to 35°C to 45°C with stirring, adding dry HCl in cyclopentyl methyl ether, and raising the temperature again to 50°C to 55°C, adding additional n-heptane as an anti-solvent, optionally adding seed crystals of crystalline obicetrapib HCl (such as in the form of a seed crystal slurry prepared as described herein), cooling to a temperature between 10°C and 15°C, then crystallizing crystalline obicetrapib HCl from the system with stirring for at least 12 hours and then filtering, washing one or more times with a mixture of cyclopentyl methyl ether and n-heptane, and drying under vacuum or the like. In some embodiments, the amount of toluene is significantly higher.

[0110] In some embodiments, the crystalline obicetrapib HCl from step (i) is isolated in step (ii). In some embodiments, the isolated crystalline obicetrapib HCl has a purity of 98% or higher, such as 98.5% or higher, 99% or higher, 99.5% or higher, 99.7% or even higher.

[0111] Another embodiment of the present disclosure relates to crystalline obicetrapib HCl obtainable or obtained by the methods defined herein.

[0112] Yet another embodiment of the present disclosure is directed to HCl obicetrapib, including crystalline obicetrapib HCl.

[0113] In some embodiments, crystalline obetrapib HCl is stored at a controlled room temperature and under a nitrogen atmosphere, protected from moisture, preventing the formation of amorphous solids, such as from desolvation. Method for preparing amorphous calcium salts of obetrapib, such as amorphous obetrapib hemicalcium - Steps (iii)-(iv) from Aspects (i)-(iv)

[0114] In some embodiments of a method for preparing amorphous calcium salts of obetrapib, such as amorphous obetrapib hemicalcium, the method includes steps (iii)-(iv), a step of preparing an amorphous calcium salt of obetrapib from crystalline obetrapib HCl isolated in step (ii), and a step of isolating an amorphous calcium salt of obetrapib, such as amorphous obetrapib hemicalcium.

[0115] In some embodiments of the method for isolating an amorphous calcium salt of obetrapib by step (iv), the amorphous calcium salt of obetrapib is in the form of amorphous obetrapib hemicalcium: [Chemical formula] as follows.

[0116] In some embodiments of a method for preparing obetrapib, step (iii) is as follows: (iii-1) Converting crystalline obetrapib HCl of step (ii) in an organic solvent to obtain obetrapib; (iii-2) Treating obetrapib in the organic solvent with aqueous sodium hydroxide to form the sodium salt of obetrapib; and (iii-3) Treating the sodium salt of obetrapib with aqueous calcium chloride to form amorphous obetrapib hemicalcium including the compounds in steps (iii-1) and (iii-2) are not isolated.

[0117] Thus, in some embodiments, step (iii-1) is as follows: (aa) Preparing crystalline obcetrapib HCl as isolated in step (ii); (bb) Dissolving crystalline obcetrapib HCl in a mixture of water and isopropyl acetate while stirring. In some embodiments, step (bb) is carried out at a temperature between 15 °C and 25 °C; (cc) Separating the phases and subjecting the resulting organic phase to one or more subsequent washings with water, wherein after each washing step, the aqueous phase is separated to obtain a washed organic phase; and (dd) Distilling the washed organic phase obtained from step (cc) two or more times at a temperature of 50 °C or less (such as 30 °C or less) (with the addition of ethanol in the middle) to obtain an ethanol solution of obcetrapib; which comprises.

[0118] In some embodiments, step (iii-2) is as follows: (ee) Adding an aqueous NaOH solution to the solution obtained in step (dd) and stirring the resulting mixture for at least 4 hours at a temperature between 20 °C and 25 °C, etc., to obtain a solution of the sodium salt of obcetrapib; and (ff) Optionally filtering the solution obtained in step (ee); which comprises.

[0119] In some embodiments, step (iii-3) is as follows: (gg) Preparing a CaCl2 solution by adding deionized water to CaCl2 while stirring, then adding ethyl acetate as a co-solvent and stirring the resulting mixture for 10 - 30 minutes; Cool the CaCl₂ solution obtained in step (gg) to a temperature of 8°C to 12°C, and at said temperature, with stirring, add it via a filter to the solution obtained in step (ff) or (ee). (ii) Stir the slurry obtained from step (hh) for about 1 to about 10 hours. In some embodiments of step (ii), the stirring is carried out at a temperature between 8°C and 12°C. (jj) Isolate the solid from the slurry obtained in step (ii) by filtration. In some embodiments of step (jj), the isolation is carried out at a temperature between 8°C and 12°C. (kk) Wash the filtration residue obtained in step (jj) one or more times with water. In some embodiments of step (kk), the washing is carried out at a temperature between 8°C and 12°C, and (ll) Dry the washed residue obtained in step (kk) at a temperature of 40°C to 50°C, under vacuum, for more than 16 hours (such as 50 hours, 100 hours, 150 hours or 200 hours or even longer) to obtain amorphous obicetrapib hemicalcium (sometimes also referred to herein as Compound 3). comprises.

[0120] In some embodiments, the amorphous obicetrapib hemicalcium is subjected to subsequent reworking procedures. In some embodiments, the amorphous obicetrapib hemicalcium is dissolved in ethanol (such as ethanol with twice the weight of the amorphous obicetrapib hemicalcium) at a temperature of 25°C to 50°C, then cooled to 10°C to 15°C, then filtered and put into a mixture of an aqueous calcium chloride solution and ethyl acetate that is also cooled to 10°C to 15°C, then filtered and washed with water, and further reworked by drying under vacuum at 45°C or less for 20 hours or longer.

[0121] In some embodiments of step (iv), amorphous obeticholic acid hemicalcium is isolated with a purity of 95% or higher, such as 95.5% or higher, 96% or higher, 96.5% or higher, 97% or higher, 97.5% or higher, 98% or higher, 98.5% or higher, 99% or higher, 99.1% or higher, 99.2% or higher, 99.3% or higher, 99.4% or higher, 99.5% or higher, 99.6% or higher, 99.7% or higher, 99.8% or higher, or 99.9% or higher.

[0122] In some embodiments, the amorphous obeticholic acid hemicalcium is subjected to a milling process. In some embodiments, the milling process is adapted to enable the production of micron-sized amorphous obeticholic acid hemicalcium (e.g., parameters such as feed rate, venturi pressure, and milling pressure are adapted). Method for preparing obeticholic acid - Step (a) from aspects (a)-(d)

[0123] In step (a) of the method for preparing obeticholic acid according to the present disclosure, the compound of formula (II) or a salt thereof is coupled with the compound of formula (III) to obtain a compound of formula (IV) (e.g., as described herein, X 1 is a leaving group and Y 1 is a protecting group).

Chemical formula

[0124] Step (a) of the subject method is the compound of formula (II) (2R,4S)-4-amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline or a salt thereof:

Chemical formula

[0125] The compound of formula (II) can be obtained, for example, using the methods disclosed in WO2016 / 024858A1 or WO2007 / 116922A1, the entire contents of both of which are incorporated herein by reference. In some embodiments, the compound of formula (II) can be obtained from the corresponding stable salts and can be obtained in pure solid form. The solid form can be amorphous or crystalline. In some embodiments, the compound of formula (II) is obtained from the corresponding crystalline salt.

[0126] In some embodiments, the compound of formula (II) obtained in step (a) is a salt of formula (IIA) or (IIB):

Chemical formula

[0127] In some embodiments, the compound of formula (II) obtained in step (a) is a salt of formula (IIA). In some embodiments, the compound of formula (IIA) is used directly in the coupling reaction with the compound of formula (III) without performing a salt decomposition step.

[0128] In some embodiments, the compound of formula (II) obtained in step (a) is a salt of formula (IIB). In some embodiments, the compound of formula (IIB) is used directly in the coupling reaction with the compound of formula (III) without performing a salt decomposition step.

[0129] In some embodiments, the compound of formula (II) in step (a) is obtained from a salt of formula (IIA) or (IIB). In some embodiments, prior to the coupling reaction of step (a), the following: (Pre-a1) A compound of formula (IIA) or (IIB):

Chemical formula

[0130] In some embodiments, the compound of formula (II) in step (a) is obtained from a salt of formula (IIA). In some embodiments, the compound of formula (II) in step (a) is obtained from a salt of formula (IIB).

[0131] In some embodiments, the salt of formula (IIA) or (IIB) is an anion A selected from sulfonate ions (e.g., besylate ion, tosylate ion, napsylate ion, camsylate ion, esylate ion, edisy late ion or mesylate ion), sulfate ions (e.g., methyl sulfate ion), halogens (e.g., chloride ion, iodide ion or bromide ion), acetate ion, aspartate ion, benzoate ion, bicarbonate ion, bitartrate ion, carbonate ion, citrate ion, decanoate ion, fumarate ion, gluceptate ion, gluconate ion, glutamate ion, glycolate ion, hexanoate ion, hydroxynaphthoate ion, isethionate ion, lactate ion, lactobionate ion, malate ion, maleate ion, mandelate ion, mucate ion, nitrate ion, octanoate ion, oleate ion, pamoate ion, pantothenate ion, phosphate ion, polygalacturonate ion, propionate ion, salicylate ion, stearate ion, succinate ion, tartrate ion and theocurate ion m- selected from salts with.

[0132] In some embodiments, the salt of formula (IIA) or (IIB) is an anion A selected from chloride ion, bromide ion, bitartrate ion, sulfate ion and sulfonate ionm- It is selected from salts with

[0133] In some embodiments, the salt of formula (IIA) or (IIB) is an anion A selected from chloride ion, bromide ion, bitartrate ion, and mesylate ion m- It is selected from salts with

[0134] In some embodiments of the salt of formula (IIA) or (IIB), m is 1.

[0135] In some embodiments, the salt is of formula (IIA), and the anion A m- is mesylate ion and m is 1. The mesylate (MSA) salt (also referred to herein as Compound 1A shown below) can be obtained by the methods disclosed in WO2016 / 024858A1 or WO2007 / 116922A1, the entire disclosures of which are incorporated herein by reference.

Chemical formula

[0136] In some embodiments, the salt decomposition of the compound of formula (IIA) or (IIB) in step (pre-a2) is carried out in a mixture of an aqueous sodium hydroxide solution and an organic solvent selected from toluene, dichloromethane, cyclopentyl methyl ether, isopropyl ether, t-butyl methyl ether, ethyl acetate, isopropyl acetate, methyl ethyl ketone, methyl isobutyl ketone, chlorobenzene, and combinations thereof. Then, the mixture is heated, then cooled, and the system is phase-separated to separate the aqueous phase. In some embodiments, the solvent is toluene. In some embodiments, the reaction mixture is heated to a temperature between 45 °C and 60 °C and then cooled to a temperature between 15 °C and 40 °C.

[0137] In some embodiments, after separating the aqueous phase, the obtained organic phase is subjected to one or more steps of washing with water, and after each step of washing with water, separation of the aqueous phase such as one or more steps of washing with an aqueous sodium chloride solution follows, then the aqueous phase is separated, and then one or more steps of washing with deionized water follow, and again, separation of the aqueous phase continues. Next, the obtained washed organic phase is optionally subjected to distillation to reduce the water content to less than 1000 ppm based on the weight of the solution. Alternatively, in some embodiments, a small amount of water still remains in the organic phase together with the compound of formula (II), and the subsequent coupling with the compound of formula (III) proceeds in the presence of this small amount of water.

[0138] In some embodiments, the salt decomposition reaction in step (pre2a) is carried out on the mesylate (compound 1A) at a temperature between 45 °C and 60 °C in a mixture of an aqueous sodium hydroxide solution and toluene, then this mixture is cooled to a temperature between 15 °C and 25 °C, and the system is phase-separated to separate the aqueous phase. Next, the toluene phase obtained after separating the aqueous phase is optionally subjected to one or more steps of washing with an aqueous sodium chloride solution, then the aqueous phase is separated, and then one or more steps of washing with deionized water follow, and again, separation of the aqueous phase follows, and then the obtained washed toluene phase is subjected to distillation under reduced pressure at a temperature between 50 °C and 65 °C to reduce the water content to less than 1000 ppm based on the weight of the total amount of the solution. Alternatively, a small amount of water still remains in toluene together with the compound of formula (II), and the subsequent coupling reaction with the compound of formula (III) proceeds in the presence of this small amount of water.

[0139] As outlined above, in step (a), the compound of formula (II) or a salt thereof (e.g., a compound of formula (IIA) or (IIB) such as mesylate 1A) is coupled with the compound of formula (III) to obtain the compound of formula (IV). In some embodiments, the method is carried out in an organic solvent.

[0140] The coupling partner of formula (III) in step (a) contains a leaving group (X 1 ). It is understood that any convenient leaving group for X 1 can be used in the present disclosure. In some embodiments, the leaving group (X 1 ) in the compound of formula (III) is selected from halogen, carbamate and substituted sulfonyloxy groups. In some embodiments, the leaving group (X 1 ) in the compound of formula (III) is a sulfonyloxy group selected from methanesulfonyloxy group, p-toluenesulfonyloxy group or trifluoromethanesulfonyloxy group. In some embodiments, the leaving group (X 1 ) is carbamate. In some embodiments, the leaving group (X 1 ) is halogen. In certain embodiments, the halogen is chloride. The coupling partner of formula (III) in step (a) also contains a protecting group (Y 1 ). The term "protecting group" refers to any group that, when attached to a functional group such as the carboxylic acid moiety of a compound (including its intermediates), prevents a reaction from occurring at that functional group, and this protecting group can be removed by conventional chemical steps or enzymatic steps to re-establish this functional group, for example, the carboxylic acid moiety. The specific removable protecting groups used are not critical. Examples of carboxylic acid protecting groups include t-butyl ester, methyl ester, ethyl ester, benzyl ester, allyl ester, 1,1-diethylallyl ester, 2,2,2-trifluoroethyl ester, phenyl ester, 4-methoxybenzyl ester, silyl ester, orthoester, esters of 2,6-disubstituted phenols (e.g., 2,6-dimethylphenol) and other conventional substituents, as well as any other group that is chemically introduced into the carboxylic acid group or a similar functional group and can be selectively removed later, either chemically or enzymatically, under mild conditions compatible with the properties of the product. Any convenient protecting group for the carboxylic acid moiety (e.g., an ester group) is Y 1It will be understood that the selection of an appropriate protecting group can be readily determined by one skilled in the art. Suitable groups for such purposes are described in Protective Groups in Organic Synthesis, 4 by T.W. Greene and P.G.M.Wuts. th Protecting groups (Y) are discussed in standard textbooks in the field of chemistry, such as Protecting Group Chemistry, 1st Ed. by Jeremy Robertson (Oxford University Press, 2000); and Protecting Group Chemistry, 8th Ed. by Michael B. Smith (Wiley-Interscience Publication, 2001). In some embodiments, protecting groups (Y 1 ) is selected from alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, allyl groups, substituted allyl groups, and silyl groups. In some embodiments, the protecting group (Y 1 ) is selected from t-butyl, methyl, ethyl, benzyl, allyl, substituted allyl, 2,2,2-trifluoroethyl, phenyl, 4-methoxybenzyl ester, 2,6-disubstituted phenol, and silyl groups. In some embodiments, the protecting group (Y 1 ) is a t-butyl group. In some embodiments, the compound of Formula (III) has the following structure 1B: [ka] is a compound of

[0141] In some embodiments of the coupling reaction of step (a), the solvent is selected from toluene, t-butanol, 1,4-dioxane, xylene, N-methyl-2-pyrrolidone, dimethylformamide, water, tetrahydrofuran, and combinations thereof. In some embodiments, the solvent is a mixture of the organic solvent toluene and the organic co-solvent t-butanol.

[0142] If steps (pre-a1) and (pre-a2) are carried out before step (a), the compound of formula (II) is already present in the required solvent, because the same effective solvent is used in steps (pre-a2) and (a), or because a solvent exchange occurs in step (pre-a2). If necessary, additional organic solvents, and for example organic co-solvents, can be added in step (a). As will be understood by those skilled in the art, organic co-solvents can also be added during the solvent exchange in step (pre-a2). In some embodiments, steps (pre-a1) and (pre-a2) are carried out before step (a), and the compound of formula (II) is present in toluene.

[0143] The coupling reaction in step (a) is usually a catalytic reaction. In some embodiments, the reaction is a palladium-catalyzed coupling reaction in the presence of a base. Suitable examples of palladium catalysts are, for example, tris(dibenzylideneacetone) dipalladium and Pd(II) acetate. Suitable bases include organic bases (such as sodium t-butoxide and potassium t-butoxide) and inorganic bases (such as K3PO4, K3PO4·H2O, sodium carbonate, potassium carbonate, cesium carbonate, LiHMDS, NaHMDS, KOH and NaOH).

[0144] In many embodiments, anhydrous K3PO4 is used as the base. In a number of such embodiments, 90% of the particles have a particle size distribution smaller than between about 140 and about 307 microns, including between about 160 and about 290 microns and between about 180 and about 220 microns and between about 200 and about 210 microns, including between about 140 and about 170 microns. In some embodiments, 90% of the particles are less than 205 microns.

[0145] In these and other embodiments, 50% of the particles are between about 35 and about 173 microns, including between about 35 and about 40 microns, or smaller.

[0146] In these and other embodiments, 10% of the particles are between about 7 and about 74 microns, including between about 7 and about 10 microns.

[0147] In some embodiments, the compound of formula (II) reacts with the compound of formula (III) in step (a) using a palladium catalyst and a base in a solvent (e.g., an organic solvent). In some embodiments, the reaction mixture further comprises a ligand.

[0148] In some embodiments, the compound of formula (IIA) or (IIB) reacts with the compound of formula (III) in step (a) using a palladium catalyst and a base in a solvent (e.g., an organic solvent). In some embodiments, the reaction mixture further comprises a ligand.

[0149] In some embodiments, the salt-decomposed compound of formula (II) reacts with the compound of formula (III) in step (a) using either Pd(II) acetate and (S)-BINAP [(S)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl] or rac-BINAP as a ligand in a solvent (e.g., an organic solvent). In some embodiments, (S)-BINAP is used as the ligand and the base is selected from sodium t-butoxide, potassium t-butoxide, K3PO4 anhydrous, K3PO4·H2O, sodium carbonate, potassium carbonate, cesium carbonate, LiHMDS, NaHMDS, KOH, and NaOH.

[0150] In some embodiments, the salt of formula (IIA) or (IIB) reacts with the compound of formula (III) in step (a) in a solvent (e.g., an organic solvent) using palladium(II) acetate, and either (S)-BINAP [(S)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl], (R)-BINAP [(S)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl] or rac-BINAP as a ligand. In some embodiments, (S)-BINAP is used as the ligand and the base is selected from sodium t-butoxide, potassium t-butoxide, K3PO4 anhydrous, K3PO4·H2O, sodium carbonate, potassium carbonate, cesium carbonate, LiHMDS, NaHMDS, KOH and NaOH. In some embodiments, the salt of formula (IIA) is Compound 1A which is a mesylate salt.

[0151] In some embodiments, the reaction in step (a) is carried out at a temperature of 70 °C to 80 °C for 2 hours or longer, optionally under a nitrogen atmosphere.

[0152] In some embodiments, the compound of formula (II) or the salt of formula (IIA) in step (a) in a mixture of the organic solvent toluene and the organic co-solvent t-butanol under a nitrogen atmosphere, using palladium(II) acetate as a catalyst, (S)-BINAP as a ligand, and K3PO4 anhydrous or K3PO4·H2O as a base, for 2 hours or longer at a temperature between 70 °C and 80 °C, with the compound of formula (III) (X 1 is Cl and Y 1 is t-butyl) reacts.

[0153] In some embodiments, the step of washing one or more times with water comprises washing one or more times with water, preferably deionized water, then separating the aqueous phase, and subsequently washing one or more times with an aqueous HCl solution, then separating the aqueous phase, subsequently washing one or more times with an aqueous sodium chloride solution, then separating the aqueous phase, and finally washing one or more times again with deionized water, then separating the aqueous phase.

[0154] In step (a), when t-butanol is used as the organic co-solvent, this organic co-solvent is removed from the organic phase during the washing step.

[0155] If step (a) is carried out in an organic solvent different from the solvent used in step (b), the organic solvent used in step (a) is exchanged in step (a) for the organic solvent applied in step (b), so that the compound of formula (IV) remains in solution.

[0156] In some embodiments where the (organic) solvents used in steps (a) and (b) are different, at least a portion of the (organic) solvent used in step (a) is evaporated, for example by using distillation under reduced pressure, and the organic solvent of step (b) is added, so that the compound of formula (IV) remains in solution during the solvent exchange. This process can be carried out by continuously evaporating the (organic) solvent used in step (a) and continuously adding the organic solvent of step (b) until, for example, the amount of the (organic) solvent used in step (a) is less than a certain threshold value relative to the total amount of the solvent. Alternatively, this process can be carried out batchwise by evaporating a portion of the (organic) solvent used in step (a) and subsequently adding a portion of the organic solvent used in step (b) more than once until the amount of the (organic) solvent used in step (a) is less than a certain threshold value relative to the total amount of the solvent.

[0157] In some embodiments, the solvent used in step (a) is a mixture of the organic solvent toluene and the organic co-solvent t-butanol. The t-butanol is removed from the organic phase containing the compound of formula (IV) during the washing step.

[0158] In some embodiments of step (a), the toluene, which is the residual organic solvent, is exchanged with acetonitrile by distilling off a portion of the toluene while adding acetonitrile in portions at a temperature between 50 °C and 65 °C under reduced pressure in two or more steps to obtain a solvent mixture containing less than about 20% by weight of toluene based on the combined weight of the solvents, so that the compound of formula (IV) remains in solution. In some embodiments of the compound of formula (IV), Y 1 is t-butyl. Method for preparing obicetrapib - step (b) from aspects (a) to (d)

[0159] In step (b) of the method for preparing the compound of formula (I) according to the present disclosure, the compound of formula (IV) is converted to a carbamate of formula (V) in an organic solvent and subsequently isolated as a solid salt of formula (VI) (Y 1 is, for example, a protecting group as described herein).

Chemical formula

[0160] In some embodiments, the organic solvent used in step (b) is selected from acetonitrile, chlorobenzene, toluene, N-methyl-2-pyrrolidone, xylene, 1,4-dioxane, ethyl acetate, isopropyl acetate, methyl ethyl ketone, methyl isobutyl ketone, dichloromethane, t-butyl methyl ether, and combinations thereof. In some embodiments, the organic solvent is acetonitrile or a mixture of chlorobenzene and dichloromethane.

[0161] As described previously in this specification, the compound of formula (IV) is already obtained in the organic solvent used in step (b) in step (a), because either the same organic solvent is used in steps (a) and (b) or a solvent exchange is carried out in step (a). In some embodiments of the compounds of formula (IV), (V) and (VI), Y 1 is t-butyl.

[0162] In some embodiments, the organic solvent used in step (b) is a mixture of acetonitrile and toluene, wherein toluene is less than about 20% by weight relative to the combined weight of the organic solvents.

[0163] In some embodiments, the conversion of the compound of formula (IV) to the corresponding carbamate having formula (V) in step (b) is carried out in acetonitrile containing less than about 20% by weight of toluene, at a temperature between 10 °C and 20 °C, in the presence of pyridine, using an excess of ethyl chloroformate, relative to the combined weight of the organic solvents.

[0164] When step (b) is carried out in an organic solvent different from the organic solvent used in step (c), the organic solvent used in step (b) is exchanged in step (b) for the organic solvent applied in step (c), such that the compound of formula (V) remains in solution.

[0165] In some embodiments where the organic solvents used in steps (b) and (c) are different, at least a portion of the organic solvent used in step (b) is evaporated, such as by distillation under reduced pressure, and the organic solvent of step (c) is added, so that the compound of formula (V) remains in solution during the organic solvent exchange. This process can be carried out by continuously evaporating the organic solvent used in step (b) and continuously adding the organic solvent of step (c) until, for example, the amount of the organic solvent used in step (b) is less than a certain threshold value with respect to the total amount of the organic solvent. Alternatively, this process can be carried out batchwise by evaporating a portion of the organic solvent used in step (b) and then adding a portion of the organic solvent used in step (c) more than once until, for example, the amount of the organic solvent used in step (b) is less than a certain threshold value with respect to the total amount of the organic solvent.

[0166] The resulting mixture is preferably treated one or more times with an aqueous solution of sodium chloride and / or HCl, then the aqueous phase is separated, and subsequently, the mixture is treated one or more times with an aqueous bicarbonate solution, and then the aqueous phase is separated.

[0167] In some embodiments, the conversion of the compound of formula (IV) to the corresponding carbamate of formula (V) in step (b) is carried out in acetonitrile containing an excess of ethyl chloroformate in the presence of pyridine at a temperature between 10 °C and 20 °C. This solvent is exchanged for isopropyl acetate in step (b) by distilling off a portion of the acetonitrile in two or more steps at a temperature of 60 °C or less under reduced pressure in such an amount that a solution of the compound of formula (V) in isopropyl acetate is obtained while adding isopropyl acetate in the middle. The above solution is treated one or more times with an NaCl / HCl aqueous solution, then the aqueous phase is separated, and subsequently, the mixture is treated one or more times with an aqueous bicarbonate solution, and then the aqueous phase can be separated.

[0168] Next, the compound of formula (V) dissolved in an organic solvent is converted to the corresponding salt with formula (VI) (A n- is an anion and n is an integer from 1 to 3). Next, the solid form of the salt with formula (VI) is isolated as a solid form.

[0169] In some embodiments, the salt of formula (VI) is a sulfonate ion (e.g., besylate ion, tosylate ion, napsylate ion, camsylate ion, esylate ion, edisylate ion, and mesylate ion), sulfate ion (e.g., methyl sulfate ion), halogen, acetate ion, aspartate ion, benzoate ion, bicarbonate ion, bitartrate ion, carbonate ion, citrate ion, decanoate ion, fumarate ion, gluceptate ion, gluconate ion, glutamate ion, glycolate ion, hexanoate ion, hydroxynaphthoate ion, isethionate ion, lactate ion, lactobionate ion, malate ion, maleate ion, mandelate ion, mucate ion, nitrate ion, octanoate ion, oleate ion, pamoate ion, pantothenate ion, phosphate ion, polygalacturonate ion, propionate ion, salicylate ion, stearate ion, succinate ion, tartrate ion, and theanate ion; an anion A n- selected from salts thereof.

[0170] In some embodiments, the salt of formula (VI) is an anion A n- selected from salts with chloride ion, bromide ion, bitartrate ion, sulfate ion, and sulfonate ion.

[0171] In some embodiments, the salt of formula (VI) is an anion A n- selected from salts with chloride ion, bromide ion, bitartrate ion, and mesylate ion.

[0172] In some embodiments, the salt form of formula (VI) is a mesylate salt, including its crystalline mesylate salt, Compound 1D:

Chemical formula

[0173] In some embodiments of the salts of formula (VI), n is 1.

[0174] The organic solvent used for the conversion from formula (V) to (VI) is not particularly limited. In some embodiments, it is selected from cyclopentyl methyl ether, isopropyl ether, t-butyl methyl ether, ethyl acetate, isopropyl acetate, and combinations thereof. In some embodiments, isopropyl acetate, or a mixture containing dichloromethane, n-heptane, and isopropyl alcohol, such as a mixture of dichloromethane, n-heptane, and isopropyl alcohol and chlorobenzene, is used. It should be noted that the compound of formula (V) has already been obtained in an organic solvent for the solvent exchange described hereinbefore.

[0175] Therefore, in some embodiments, the organic solvent used for the conversion of the compound of formula (V) to its corresponding salt of formula (VI) is selected from cyclopentyl methyl ether, isopropyl ether, t-butyl methyl ether, ethyl acetate, isopropyl acetate, and combinations thereof, containing less than about 20% by weight of toluene and less than about 7% by weight of acetonitrile based on the combined weight of the solvents. In some embodiments, the solvent is a mixture of isopropyl acetate, toluene, and acetonitrile containing less than about 20% by weight of toluene and less than about 7% by weight of acetonitrile based on the combined weight of the solvents.

[0176] In some embodiments, it is preferred to add an organic cosolvent different from the organic solvent already used in step (b). Exemplary organic cosolvents are selected from cyclopentyl methyl ether, isopropyl ether, t-butyl methyl ether, ethyl acetate, isopropyl acetate, such as methyl t-butyl ether, and combinations thereof. As will be appreciated by those skilled in the art, the need and advantages of using an organic cosolvent depend on the particular organic solvent already used in step (b). In certain cases, the use of a cosolvent may be omitted.

[0177] In some embodiments, the organic solvent for the conversion of the compound of formula (V) to its corresponding salt of formula (VI) comprises isopropyl acetate and methyl t-butyl ether as the organic cosolvent.

[0178] Subsequently, an acid is added to form the salt of formula (VI) as defined above. In some embodiments, the acid is selected from ditartartic acid, sulfuric acid, sulfonic acid, hydrogen bromide and hydrogen chloride. In some embodiments, the acid is methanesulfonic acid. In embodiments where the salt of formula (VI) can be obtained in crystalline form, some of the acid required to form the salt of formula (VI) can be added before crystallization and some can be added during crystallization.

[0179] The solid form of the salt of formula (VI) is isolated by crystallization, filtration, optional washing of the filter residue one or more times, and drying, when the salt of formula (VI) can be obtained in crystalline form.

[0180] In some embodiments, the compound of formula (V) is converted to its corresponding mesylate according to formula (VI) with methanesulfonic acid in an organic solvent mixture of isopropyl acetate and methyl t-butyl ether containing less than about 20% by weight of toluene and less than 7% by weight of acetonitrile relative to the combined weight of the organic solvents, and then the mesylate is crystallized from the organic solvent with compound 1D, followed by filtration, optional washing of the filter residue one or more times, and drying.

[0181] In some embodiments, where the salt according to formula (VI) can be obtained in crystalline form, crystallization is induced by adding seed crystals of the salt according to formula (VI).

[0182] In some embodiments, where the salt according to formula (VI) can be obtained in crystalline form, the step of crystallizing the salt according to formula (VI) to obtain the crystalline form of the salt according to formula (VI) is carried out by adding the acid necessary to form the salt and stirring the resulting mixture at a temperature of 20 °C to 25 °C for more than 60 minutes, crystallizing while stirring at a temperature between 15 °C and 25 °C for more than 120 minutes, then subjecting the resulting slurry to vacuum filtration (the filtration residue is washed one or more times with the same organic solvent used to crystallize the salt according to formula (VI)), and vacuum drying the crystalline form of the salt according to formula (VI).

[0183] In an embodiment, the present invention relates to a salt according to formula (VI), where A n- is an anion and n is an integer from 1 to 3. In some embodiments, the compound is the crystalline methanesulfonic acid (MSA) salt of formula (VI) (e.g., Compound 1D described herein).

[0184] In some embodiments, the step of crystallizing the mesylate salt of formula (VI) from an organic solvent mixture of isopropyl acetate and methyl t-butyl ether to obtain the crystalline form of the mesylate salt by Compound 1D is carried out by adding methanesulfonic acid necessary to form the salt, stirring the resulting mixture at a temperature between 15 °C and 25 °C (e.g., 20 °C) for longer than 60 minutes, and then crystallizing while stirring at a temperature between 15 °C and 25 °C for more than 120 minutes. The resulting slurry is subjected to vacuum filtration, and the filtration residue is washed one or more times with a mixture of isopropyl acetate and methyl t-butyl ether and dried under vacuum to obtain the crystalline form of the mesylate salt by Compound 1D.

[0185] In some embodiments, the compound of formula (VI) is obtained in a yield of at least 70% based on the number of moles of the compound of formula (II). In some embodiments, the compound of formula (VI) is obtained with a purity of 99% or higher, such as 99.1% or higher, 99.2% or higher, 99.3% or higher, 99.5% or higher, or even higher purity. Method for Preparing Obeticholic Acid - Step (c) from Aspects (a) to (d)

[0186] In step (c) of the method according to the present disclosure, when an isolated salt of formula (VI) or a salt-decomposed derivative thereof (e.g., a compound according to formula (V)) is alkylated with a compound of formula (VII), a compound of formula (VIII):

Chemical formula

[0187] In some embodiments of step (c), an isolated solid form of a salt according to formula (VI), such as a crystalline form of the salt according to formula (VI) (such as Compound 1D which is a crystalline mesylate), when reacted directly with a compound of formula (VII) in an organic solvent, forms a compound of formula (VIII) (i.e., without a salt decomposition step).

[0188] In some embodiments of step (c), an isolated solid form of a salt according to formula (VI), such as a crystalline form of the salt according to formula (VI) (such as Compound 1D which is a crystalline mesylate), is salt-decomposed and then reacted with a compound of formula (VII) in an organic solvent to form a compound of formula (VIII). Salt decomposition of the compound of formula (VI) results in a compound according to formula (V).

[0189] When the compound of formula (VI) is subjected to a salt decomposition step, the salt decomposition process and the subsequent reaction with the compound of formula (V) are carried out in the same organic solvent. In some embodiments, the organic solvent is selected from xylene, n - hexane, toluene, heptane (mixture of isomers), n - heptane, dichloromethane, chlorobenzene and combinations thereof. In some embodiments, the organic solvent is toluene or n - heptane.

[0190] In some embodiments, step (c) is carried out in the presence of a base. In some embodiments, step (c) is carried out in the presence of a solid - liquid phase transfer catalyst.

[0191] In some embodiments, the base is selected from alkali metal hydrides, alkali metal hydroxides, alkaline earth metal hydroxides, alkali metal alkoxides, alkali metal carbonates, alkali metal bicarbonates and amines. In some embodiments, the base is selected from alkali metal alkoxides. In some embodiments, the base is sodium t - pentoxide, or a mixture of sodium t - butoxide and potassium t - butoxide.

[0192] In some embodiments, the solid - liquid phase transfer catalyst is selected from t - butylammonium hydrogen sulfate, tetra - n - butylammonium bromide, tetra - n - butylammonium iodide, crown ethers and combinations thereof. In some embodiments, the catalyst is t - butylammonium hydrogen sulfate.

[0193] In some embodiments, the reaction of the compound of formula (V) or (VI) with the compound of formula (VII) is carried out at a temperature between 0 °C and 25 °C (such as 5 °C to 20 °C).

[0194] The coupling partner of formula (VII) in step (c) is a leaving group X 2 which is included. It is understood that any convenient leaving group can be used in the present disclosure with respect to X 2 . In some embodiments, the leaving group X 2is selected from a halogen and a substituted sulfonyloxy group. In some embodiments, the leaving group X in the compound of formula (VII) 2 is a substituted sulfonyloxy group selected from a methanesulfonyloxy group, a p-toluenesulfonyloxy group or a trifluoromethanesulfonyloxy group. In some embodiments, the leaving group X 2 is a halogen. In certain embodiments, the halogen is bromide. In some embodiments, the compound of formula (VII) has the following structure 1E:

Chemical formula

[0195] In some embodiments, the salt decomposition of the compound of formula (VI) and the subsequent reaction with the compound of formula (VII) in step (c) are carried out in toluene as an organic solvent in the presence of a base and a catalyst at a temperature of 5°C to 25°C. In some embodiments, the salt decomposition of the compound of formula (VI) and the subsequent reaction with the compound of formula (VII) in step (c) are carried out at a temperature between 5°C and 25°C, with stirring for about 1 to 8 hours, in toluene as an organic solvent in the presence of sodium t-pentoxide as a base and t-butylammonium hydrogen sulfate as a catalyst. In some embodiments of the compound of formula (VI), Y 1 is t-butyl.

[0196] In some embodiments, the alkylation of the compound of formula (VI) with the compound of formula (VII) in step (c) (i.e., without an additional salt decomposition step) is carried out in toluene as an organic solvent in the presence of a base and a catalyst at a temperature of 5°C to 25°C. In some embodiments, the alkylation of the compound of formula (VI) with the compound of formula (VII) in step (c) is carried out at a temperature between 5°C and 25°C, with stirring for about 1 to 8 hours, in toluene as an organic solvent in the presence of sodium t-pentoxide as a base and t-butylammonium hydrogen sulfate as a catalyst.

[0197] In some embodiments, step (c) comprises the steps of preparing crystalline 1D, decomposing this compound by salt, and reacting the compound after salt decomposition with a compound of formula (VII) (X 2 is Br) in toluene as an organic solvent at a temperature between 5°C and 25°C for about 1 to 8 hours with stirring in the presence of sodium t-pentoxide as a base and t-butylammonium hydrogen sulfate as a catalyst.

[0198] In some embodiments, step (c) comprises reacting crystalline 1D with a compound of formula (VII) (X 2 is Br) in toluene as an organic solvent at a temperature between 5°C and 25°C for about 1 to 8 hours with stirring in the presence of sodium t-pentoxide as a base and t-butylammonium hydrogen sulfate as a catalyst.

[0199] In some embodiments of step (c), the base is the last reagent added to the reaction mixture. Without being bound by any particular theory, the inventors have discovered that by adding the base as the last reagent, the equivalent numbers of both the base used in the reaction mixture and the compound of formula (VII) can be reduced. The reduction in the equivalent number of the compound of formula (VII) can, in turn, reduce the risk of carrying over impurities associated with formula (VII) into the final product.

[0200] Accordingly, step (c) results in the formation of a compound of formula (VIII) in an organic solvent. In some embodiments of the compound of formula (VIII), Y 1is t-butyl. In some embodiments, the reaction mixture is subjected to one or more water washing steps in step (c) to remove impurities, followed by separation of the aqueous phase and, optionally, one or more filtration steps as necessary, to obtain a washed reaction mixture containing the compound of formula (VIII) in an organic solvent. In some embodiments, the reaction mixture containing the compound of formula (VIII) in an organic solvent is concentrated by distilling off a portion of the organic phase to obtain a concentrated reaction mixture containing the compound of formula (VIII) in an organic solvent. In some embodiments, the organic solvent contains 30 to 40% by weight of the compound of formula (VIII) based on the weight of the reaction mixture. In some embodiments, the organic solvent contains 34 to 37% by weight of the compound of formula (VIII) based on the weight of the reaction mixture.

[0201] The one or more water washing steps, the one or more filtration steps as necessary and the concentration step are preferably combined together, thus obtaining a washed and concentrated reaction mixture containing the compound of formula (VIII) in an organic solvent. In some cases, the organic solvent contains 30 to 40% by weight of the compound of formula (VIII). In some embodiments, the organic solvent contains 34 to 37% by weight of the compound of formula (VIII) based on the weight of the reaction mixture.

[0202] In some embodiments, the one or more water washing steps include one or more washing steps with an aqueous acetic acid solution.

[0203] In some embodiments, the reaction mixture containing the compound of formula (VIII) in toluene as the organic solvent is subjected in step (c) to one or more water washing steps with an aqueous acetic acid solution, followed by a step of separating the aqueous phase, and then, usually, a step of distilling off a portion of toluene under reduced pressure at a temperature of 75°C to 90°C, to obtain a washed and concentrated reaction mixture containing 30 to 40% by weight of the compound of formula (VIII) based on the weight of the reaction mixture and containing the compound of formula (VIII) in toluene. In some embodiments, the concentrated mixture contains 34 to 37% by weight of the compound of formula (VIII) based on the weight of the reaction mixture.

[0204] When step (c) is carried out in an organic solvent different from the organic solvent used in step (d), the organic solvent used in step (c) is exchanged in step (c) for the organic solvent applied in step (d), and thus the compound of formula (VIII) remains in solution.

[0205] In some embodiments where the organic solvents used in steps (c) and (d) are different, at least a portion of the organic solvent used in step (c) is preferably evaporated using distillation under reduced pressure, and the organic solvent of step (d) is added such that the compound of formula (VIII) remains in solution during the solvent exchange. This process can be carried out by continuously evaporating the organic solvent used in step (c) until, for example, the amount of the organic solvent used in step (c) is below a certain threshold relative to the total amount of the organic solvent, and continuously adding the organic solvent of step (d). Alternatively, this process can be carried out batchwise by evaporating a portion of the organic solvent used in step (c) and subsequently adding a portion of the organic solvent used in step (d) more than once until, for example, the amount of the organic solvent used in step (c) is below a certain threshold relative to the total amount of the organic solvent. Method for preparing the compound of formula (I) - step (d) from aspects (a) to (d)

[0206] In step (d) of the method according to the present disclosure, the compound of formula (VIII) is converted to obicetrapib in a first organic solvent (Y 1 is, for example, a protecting group as described herein).

Chemical formula

[0207] The selection of the first organic solvent used in step (d) is not particularly limited. In some embodiments, the first organic solvent is neither an ether nor an ester. In some embodiments, the first organic solvent is toluene or a mixture of n-heptane and acetic acid. As previously explained herein, the compound of formula (VIII) is already obtained in the first solvent used in step (d) in step (c), because either the same organic solvent is used in steps (c) and (d) or a solvent exchange occurs in step (c).

[0208] Accordingly, in some embodiments, the first organic solvent as defined previously herein, containing 30 to 40 wt% of the compound of formula (VIII), such as 34 to 37 wt% based on the weight of the reaction mixture, is supplied in step (d).

[0209] In some embodiments, toluene containing 30 to 40 wt% of the compound of formula (VIII), such as 34 to 37 wt% based on the weight of the reaction mixture, is supplied as the first organic solvent in step (d).

[0210] Any convenient protecting group for the carboxylic acid, such as an ester moiety, can be used as Y in the compound of formula (VIII). As disclosed herein, the selection of a suitable protecting group for the carboxylic acid can be readily determined by one of ordinary skill in the art. In some embodiments of formula (VIII), the protecting group (Y 1 ) is selected from alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, allyl groups, substituted allyl groups, and silyl groups. In some embodiments of formula (VIII), the protecting group (Y 1 ) is selected from t-butyl, methyl, ethyl, benzyl, allyl, substituted allyl, 2,2,2-trifluoroethyl, phenyl, 4-methoxybenzyl ester, 2,6-disubstituted phenol, and silyl groups. In some embodiments of the compound of formula (VIII), the protecting group Y 1 ) is selected from t-butyl, methyl, ethyl, benzyl, allyl, substituted allyl, 2,2,2-trifluoroethyl, phenyl, 4-methoxybenzyl ester, 2,6-disubstituted phenol, and silyl groups. In some embodiments of the compound of formula (VIII), the protecting group Y 1is t-butyl. In some embodiments, the conversion of the compound of formula (VIII) to obicetrapib is carried out by contacting the compound of formula (VIII) with acetic acid (AcOH) and dry HCl while stirring in a first organic solvent such as toluene or a mixture of n-heptane and acetic acid. In some embodiments, the reaction mixture is heated to a temperature between 40 °C and 55 °C and the resulting mixture is maintained at this temperature while stirring for at least 3 hours.

[0211] Obicetrapib can be isolated from the resulting mixture using techniques known to those skilled in the art.

[0212] In some embodiments, in step (d), the resulting mixture containing obicetrapib is subjected to one or more water wash steps. In some embodiments, the one or more water wash steps in step (d) are carried out as follows: (AA) Cool the reaction mixture containing obicetrapib to a temperature between 15 °C and 25 °C, then add a mixture of n-heptane, acetonitrile and water, and then stir the resulting mixture at this temperature for longer than 15 minutes. (BB) Phase-separate the system obtained in step (AA) into an organic phase and an aqueous phase and separate both phases. (CC) Add a mixture of n-heptane, acetonitrile, toluene and water to the aqueous phase obtained in step (BB), and then stir the resulting system at a temperature between 15 °C and 25 °C for longer than 15 minutes. (DD) Phase-separate the system obtained in step (CC) into an organic phase and an aqueous phase and separate both phases. (EE) Combine the organic phase obtained in step (BB) and the organic phase obtained in step (DD), add water, and stir the resulting system at a temperature between 15 °C and 25 °C for longer than 15 minutes. (FF) Phase-separate the system obtained in step (EE) into an organic phase and an aqueous phase and separate both phases. Add water to the organic phase obtained in step (FF), and stir the resulting system for longer than 15 minutes at a temperature between 15 °C and 25 °C. (HH) Separate the system obtained in step (GG) into an organic phase and an aqueous phase, and separate both phases. (II) Add an aqueous solution of trisodium citrate dihydrate to the organic phase obtained in step (HH), and then stir the resulting mixture for longer than 15 minutes at a temperature between 15 °C and 25 °C. (JJ) Separate the system obtained in step (II) into an organic phase and an aqueous phase, and separate both phases. (KK) Add water to the organic phase obtained in step (JJ), and stir the resulting system for longer than 15 minutes at a temperature between 15 °C and 25 °C, and (LL) Separate the system obtained in step (KK) into an organic phase and an aqueous phase, and separate both phases.

[0213] Steps (AA) to (LL) in this embodiment result in the compound of formula (I) after washing in an organic solvent mixture containing n-heptane, acetonitrile and a first organic solvent. In some embodiments, the first solvent is toluene.

[0214] In some embodiments, the first organic solvent still does not mainly consist of cyclopentyl methyl ether, and the organic solvent mixture is exchanged with CPME in a subsequent step (MM), so that obicetrapib remains in solution.

[0215] Thus, in some embodiments, step (MM) follows step (LL), at least a portion of the solvent in the organic solvent mixture obtained in step (LL) is evaporated, such as by distillation under reduced pressure, cyclopentyl methyl ether is added, and thus, obicetrapib remains in solution during the solvent exchange. In some embodiments, the method provides a solution of obicetrapib in cyclopentyl methyl ether at a concentration between 30 and 40 wt% based on the weight of the solution. In some embodiments, the concentration of obicetrapib in cyclopentyl methyl ether is 33 to 37 wt% based on the weight of the solution, the first organic solvent is less than 1 wt% based on the weight of the solution, and n-heptane is less than 1 wt% based on the weight of the solution.

[0216] This process can be carried out by continuously evaporating the solvent in the organic solvent mixture obtained in step (LL) until, for example, the amount of a particular solvent in the organic solvent mixture is below a particular threshold relative to the total amount of the organic solvent, and continuously adding cyclopentyl methyl ether. Alternatively, this process can be carried out batchwise more than once by evaporating a portion of the solvent in the organic solvent mixture obtained in step (LL) and subsequently adding cyclopentyl methyl ether until, for example, the amount of a particular solvent in the organic solvent mixture is below a particular threshold relative to the total amount of the solvent.

[0217] In some embodiments, the first organic solvent is toluene, and after step (LL), step (MM) follows. In step (LL), at least a portion of n-heptane, acetonitrile, and toluene in the organic solvent mixture obtained is evaporated, for example, by distillation under reduced pressure (vacuum) at a temperature of 45 °C or less while adding cyclopentyl methyl ether in the middle. Thus, obicetrapib remains in the solution during the solvent exchange, resulting in a cyclopentyl methyl ether solution of obicetrapib having a concentration between 30 and 40% by weight. In some embodiments, the concentration of obicetrapib in cyclopentyl methyl is 33 to 37% by weight based on the weight of the solution, and contains less than 0.5% by weight of toluene, less than 0.5% by weight of acetonitrile, and less than 2.7% by weight of n-heptane. Method for preparing crystalline obicetrapib HCl - Steps (e)-(f) added to aspects (a)-(d)

[0218] In some embodiments of the subject method, steps (e)-(f) follow step (d), and obicetrapib is treated with HCl, such as in a suitable solvent. Such a solvent may be an aqueous solvent or an organic solvent. In some embodiments, the use of an organic solvent results in crystalline obicetrapib HCl.

[0219] In some embodiments, the organic solvent used in step (e) comprises a mixture of a solvent and an anti-solvent. In some embodiments, the solvent is selected from methanol, ethanol, isopropanol, acetic acid, acetonitrile, acetone, methyl isobutyl ketone, isopropyl acetate, tetrahydrofuran, methyl t-butyl ether, cyclopentyl methyl ether, N-methyl-2-pyrrolidone, dimethyl sulfoxide, dimethylformamide, 2-methyl-tetrahydrofuran, dichloromethane, 1,4-dioxane, 1,2-difluorobenzene, toluene, hexafluoroisopropanol, and water. In some embodiments, the anti-solvent is selected from n-heptane, n-hexane, n-pentane, and cyclohexane. In some embodiments, HCl has sufficient solubility in the anti-solvent such that the anti-solvent can be used as a suitable solvent. In some embodiments, the organic solvent used in step (e) comprises a mixture of cyclopentyl methyl ether and n-heptane. In some embodiments, the organic solvent used in step (e) further comprises toluene.

[0220] In some embodiments, step (e) includes preparing obeticholic acid in a mixture of cyclopentyl methyl ether and n - heptane, raising the temperature to between 35°C and 40°C while stirring, adding dry HCl in cyclopentyl methyl ether, and raising the temperature again to between 50°C and 55°C. Next, additional n - heptane is added as an anti - solvent. At this point, a small amount of the reaction mixture is extracted, cooled to a temperature between 10°C and 15°C, and a slurry of crystals of crystalline obeticholic acid HCl (referred to herein as the "seed crystal slurry") can be obtained in a mixture of cyclopentyl methyl ether and n - heptane. Optionally, all or part of the seed crystal slurry of crystalline obeticholic acid HCl can then be added back to the reaction mixture as seed crystals. The seed aids in nucleation but is not necessary, and thus the methods described herein can be carried out without seed crystal addition. Next, the resulting reaction mixture is cooled to a temperature between 5°C and 15°C (such as 10°C - 15°C), and then crystalline obeticholic acid HCl is crystallized from the system while stirring. In some embodiments, crystalline obeticholic acid HCl is crystallized over 12 hours or longer, then filtered (e.g., through a filter dryer), followed by optional washing one or more times with a mixture of cyclopentyl methyl ether and n - heptane, etc., and drying. In some cases, the wet filter cake of crystalline obeticholic acid HCl is dried under vacuum using step - wise temperatures of between 25°C and 30°C, 30°C and 40°C, 40°C and 50°C, and then 50°C and 55°C, such as 25°C, 35°C, 46°C, and 54°C.

[0221] Accordingly, in some embodiments, a method of preparing crystalline obeticholic acid HCl includes the addition of a seed crystal (e.g., as a seed crystal slurry). The seed crystal of crystalline obeticholic acid HCl is obtained by extracting a small amount of the reaction mixture after the addition of dry HCl in cyclopentyl methyl ether and the anti-solvent n-heptane according to step (i) described above, and cooling to a temperature between 10 °C and 15 °C to obtain a slurry of crystals of crystalline obeticholic acid HCl in cyclopentyl methyl ether and n-heptane, and can be formed as a slurry.

[0222] In some embodiments, the organic solvent used in step (e) includes a mixture of cyclopentyl methyl ether and n-heptane. Accordingly, in one embodiment, step (e) includes the steps of preparing obeticholic acid in a mixture of cyclopentyl methyl ether and n-heptane, raising the temperature to 35 °C to 45 °C while stirring, adding dry HCl in cyclopentyl methyl ether, and raising the temperature again to 50 °C to 55 °C, adding additional n-heptane as an anti-solvent, adding a seed crystal of crystalline obeticholic acid HCl (e.g., as a seed crystal slurry prepared as described herein) as needed, cooling to a temperature between 5 °C and 15 °C (such as 10 °C to 15 °C), and then crystallizing crystalline obeticholic acid HCl from this system while stirring. In some embodiments, crystalline obeticholic acid HCl is crystallized over at least 12 hours, then filtered, washed one or more times as needed with a mixture of cyclopentyl methyl ether and n-heptane, etc., and drying follows. In some embodiments, crystalline obeticholic acid HCl is dried under vacuum. In some embodiments, crystalline obeticholic acid HCl is dried in a vacuum drying cabinet at a pressure of 25 mbar and a temperature of 55 °C for 10 hours or longer. In some embodiments, after the drying procedure, crystalline obeticholic acid HCl contains less than 0.1 wt% residual cyclopentyl methyl ether.

[0223] In some embodiments described previously in this specification, step (MM) of step (d) provides a solution of obicetrapib in cyclopentyl methyl ether having a concentration between 30 and 40 wt%, such as 33 - 37 wt%, based on the weight of the solution, less than 1 wt% of the first organic solvent used in step (d), and less than 1 wt% n - heptane. In some embodiments described previously in this specification, step (MM) of step (d) provides a solution of obicetrapib in cyclopentyl methyl ether having a concentration between 30 and 40 wt%, such as 33 - 37 wt%, based on the weight of the solution, less than 1 wt% toluene, and less than 1 wt% n - heptane. These solutions may advantageously be used in step (e) after the addition of n - heptane. As will be appreciated by those skilled in the art, n - heptane may also be added in step (d).

[0224] Accordingly, in some embodiments, step (e) includes preparing a solution of obicetrapib in cyclopentyl methyl ether at a concentration between 30% and 40% by weight, such as 33% to 37% by weight, based on the weight of the solution (wherein the first organic solvent (such as toluene) used in step (d) is less than 1% by weight and n-heptane is less than 1% by weight), adding n-heptane, raising the temperature to 35°C to 45°C while stirring, adding dry HCl in cyclopentyl methyl ether, and again raising the temperature to 50°C to 55°C, adding additional n-heptane as an anti-solvent, optionally adding seed crystals of crystalline obicetrapib HCl (such as in the form of a seed crystal slurry prepared as described herein), cooling to a temperature between 5°C and 15°C (such as 10°C to 15°C), then crystallizing crystalline obicetrapib HCl from the system while stirring for at least 12 hours and then filtering, washing one or more times with a mixture of cyclopentyl methyl ether and n-heptane, and drying. In some cases, the wet filter cake of crystalline obicetrapib HCl is dried in vacuo using stepwise temperatures of 25°C to 30°C, 30°C to 40°C, 40°C to 50°C, and then 50°C to 55°C, such as 25°C, 35°C, 46°C, and 54°C.

[0225] In some embodiments, step (f) is as follows: (aa) preparing crystalline obicetrapib HCl; (bb) dissolving crystalline obicetrapib HCl in ethanol while stirring. In some embodiments, this is carried out between 15°C and 25°C; (cc) adding an aqueous NaOH solution to the solution obtained in step (bb) and stirring the resulting mixture at a temperature such as 20°C to 25°C for at least 4 hours to obtain a solution of the sodium salt of obicetrapib; (dd) optionally filtering the solution obtained in step (cc); (ee) While stirring, prepare a CaCl2 solution by adding deionized water to CaCl2, then add ethyl acetate as a co-solvent and stir the resulting mixture for 10 to 30 minutes. (ff) Cool the CaCl2 solution obtained in step (ee) to a temperature between 8 °C and 12 °C, and at said temperature, while stirring, add it via a filter to the solution obtained in step (dd) (or (cc)). (gg) Stir the slurry obtained from step (ff) for about 1 to about 10 hours. In some embodiments, the slurry is stirred at a temperature between 8 °C and 12 °C. (hh) Isolate the solid from the slurry obtained in step (gg) by filtration. In some embodiments, the isolation step is carried out at a temperature between 8 °C and 12 °C. (ii) Wash the filter residue obtained in step (hh) one or more times with water. In some embodiments, the washing is carried out at a temperature between 8 °C and 12 °C, and (jj) Dry the washed residue obtained in step (ii) under vacuum or the like at a temperature between 40 °C and 50 °C for more than 16 hours (such as 200 hours or longer) to obtain amorphous obcetrapib hemicalcium. comprises.

[0226] In some embodiments of the subject method, crystalline obcetrapib HCl is isolated in step (f) with a purity of 98% or higher, such as 98.5% or higher, 99% or higher, 99.5% or higher, or even higher.

[0227] Another embodiment of the present disclosure relates to crystalline obcetrapib HCl obtainable or obtained by the method defined herein.

[0228] Yet another embodiment of the present disclosure is directed to crystalline obcetrapib HCl.

[0229] In some embodiments, crystalline obeticholic acid HCl, including crystalline obeticholic acid HCl, is hygroscopic, so crystalline obeticholic acid HCl is stored under a controlled room temperature and nitrogen atmosphere, protected from moisture, and the formation of amorphous solids is prevented. Method for preparing amorphous obeticholic acid hemicalcium - Steps (g) - (h) added to embodiments (a) - (f)

[0230] In some embodiments of the subject method, steps (g) - (h) follow step (f), and crystalline obeticholic acid HCl is converted to amorphous obeticholic acid hemicalcium (Formula IB): [Chemical formula] .

[0231] In some embodiments, step (g), which is the preparation of amorphous obeticholic acid hemicalcium, comprises steps (g1) - (g3) described below: (g1) In an organic solvent, converting the crystalline obeticholic acid HCl of step (f) to obeticholic acid, (g2) Treating obeticholic acid in the organic solvent with aqueous sodium hydroxide to form the sodium salt of obeticholic acid, and (g3) Treating the sodium salt of obeticholic acid with aqueous calcium chloride to form amorphous obeticholic acid hemicalcium and the compounds in steps (g1) and (g2) are not isolated.

[0232] Thus, in some embodiments, step (g1) is as follows: (aa) Providing crystalline obeticholic acid HCl as defined or obtained in step (f), (bb) While stirring, dissolve crystalline obicetrapib HCl in a mixture of water and isopropyl acetate. In some embodiments, step (bb) is carried out at a temperature between 15 °C and 25 °C, (cc) A step of performing phase separation and subjecting the resulting organic phase to a step of washing with water one or more times thereafter, separating the aqueous phase following each washing step to obtain a washed organic phase, and (dd) Perform distillation two or more times (adding ethanol in the middle) on the washed organic phase obtained from step (cc) at a temperature of 50 °C or less (such as 30 °C or less) to obtain an ethanol solution of the obicetrapib compound, including. In some embodiments, step (g2) is as follows: (ee) Add an aqueous NaOH solution to the solution obtained in step (dd), stir the resulting mixture at a temperature between 20 °C and 25 °C for at least 4 hours to obtain a solution of the sodium salt of obicetrapib, and (ff) Optionally filter the solution obtained in step (ee) including.

[0233] In some embodiments, step (g3) is as follows: (gg) Prepare a CaCl2 solution by adding deionized water to CaCl2 while stirring, then add ethyl acetate as a co-solvent, and stir the resulting mixture for 10 to 30 minutes, (hh) Cool the CaCl2 solution obtained in step (gg) to a temperature of 8 °C to 12 °C, and at said temperature, add the solution obtained in step (ff) or (ee) through a filter while stirring, (ii) Stir the slurry obtained from step (hh) for about 1 to 10 hours. In some embodiments of step (ii), the stirring is carried out at a temperature between 8 °C and 12 °C, (jj) The step of isolating the solid from the slurry obtained in step (ii) by filtration. In some embodiments of step (jj), the isolation is carried out at a temperature between 8 °C and 12 °C, (kk) The step of washing the filtration residue obtained in step (jj) with water one or more times. In some embodiments of step (kk), the washing is carried out at a temperature between 8 °C and 12 °C, and (ll) The step of drying the washed residue obtained in step (kk) at a temperature of 40 °C to 50 °C for more than 16 hours (such as 50 hours, 100 hours, 150 hours or 200 hours, or even longer) under vacuum, etc., to obtain amorphous obeticholic acid hemicalcium (sometimes also referred to as compound 3 herein) comprising.

[0234] In some embodiments, step (g) is as follows: (aa) The step of preparing crystalline obeticholic acid HCl as defined or obtained in step (f), (bb) The step of dissolving crystalline obeticholic acid HCl in ethanol while stirring. In some embodiments, at between 15 °C and 25 °C, (cc) Adding an aqueous NaOH solution to the solution obtained in step (bb) and stirring the resulting mixture at a temperature of 20 °C to 25 °C for at least 4 hours, etc., to obtain a solution of the sodium salt of obeticholic acid, (dd) Optionally, the step of filtering the solution obtained in step (cc), (ee) Preparing a CaCl2 solution by adding deionized water to CaCl2 while stirring, then adding ethyl acetate as a co-solvent, and stirring the resulting mixture for 10 to 30 minutes (ff) Cooling the CaCl2 solution obtained in step (ee) to a temperature between 8 °C and 12 °C, and adding the solution obtained in step (dd) or (cc) through a filter while stirring at said temperature, (gg) Step of stirring the slurry obtained from step (ff) for about 1 to 10 hours. In some embodiments, the slurry is stirred at a temperature between 8°C and 12°C. (hh) Step of isolating the solid from the slurry obtained in step (gg) by filtration. In some embodiments, the isolation is carried out at a temperature between 8°C and 12°C. (ii) Step of washing the filter residue obtained in step (hh) with water one or more times. In some embodiments, the washing is carried out at a temperature between 8°C and 12°C, and (jj) Step of drying the washed residue obtained in step (ii) under vacuum or the like at a temperature between 40°C and 50°C for more than 16 hours (such as 50 hours, 100 hours, 150 hours or 200 hours, or longer) to obtain the amorphous hemicalcium salt of formula (IB). comprises.

[0235] In some embodiments, amorphous obeticholic acid hemicalcium salt is stored sealed at a temperature below 30°C and protected from light.

[0236] In some embodiments, amorphous obeticholic acid hemicalcium salt is subjected to subsequent reworking procedures. In some embodiments, amorphous obeticholic acid hemicalcium salt is dissolved in ethanol (such as ethanol with twice the weight of amorphous obeticholic acid hemicalcium salt) at a temperature of 25°C to 50°C, then cooled to 10°C to 15°C, then filtered and put into a mixture of an aqueous calcium chloride solution cooled to 10°C to 15°C and ethyl acetate, then filtered, washed with water, and further reworked by drying under vacuum at 45°C or less for 20 hours or longer.

[0237] In many embodiments of the present disclosure, amorphous obeticholic acid hemicalcium is processed to achieve a particle size distribution. In many embodiments, such processing is by milling. Examples of milling include hammer milling, ball milling, and jet milling. In other embodiments, spray drying may be used to achieve a particle size distribution. Thus, in some embodiments of the present disclosure, spray-dried amorphous obeticholic acid hemicalcium is obtained. An example of jet-milled amorphous obeticholic acid hemicalcium is presented in Example 14.

[0238] In many embodiments of the present disclosure, unmilled amorphous obeticholic acid hemicalcium is provided. In many embodiments of the present disclosure, milled amorphous obeticholic acid hemicalcium is provided.

[0239] In many embodiments, the particle size distribution of amorphous obeticholic acid hemicalcium is a particle size distribution such that 90% of the particles have a diameter of about 15 microns or less. In these and other embodiments, 90% of the particles have a diameter of about 14 microns or less, 13 microns or less, 12 microns or less, 11 microns or less, 10 microns or less, 9 microns or less, 8 microns or less, 7 microns or less, 6 microns or less, 5 microns or less, 4 microns or less, or 3 microns or less.

[0240] In some embodiments, 90% of the particles have a diameter between about 6 microns and 15 microns.

[0241] In these and other embodiments, the particle size distribution of amorphous obeticholic acid hemicalcium is a particle size distribution such that 50% of the particles have a diameter of about 5 microns or less, such as 4 microns or less or 3 microns or less.

[0242] In these and other embodiments, the particle size distribution of amorphous obeticholic acid hemicalcium is a particle size distribution such that 10% of the particles have a diameter of about 2 microns or less.

[0243] The amorphous obeticholic acid hemicalcium of the present disclosure can be made with high chemical purity by the methods of the present disclosure. Such levels of purity include purities greater than 98.0%, such as 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9%, or higher. The highest levels of purity, such as greater than 99.8% or 99.9% purity, are more readily achieved using methods in which crystalline obeticholic acid HCl is used as an intermediate.

[0244] As summarized above, amorphous calcium salts of obeticholic acid, including amorphous obeticholic acid hemicalcium, are also provided herein. Novel intermediates for use in the synthesis of obeticholic acid and its salts are also provided.

[0245] The term "pharmaceutically acceptable" indicates that a substance has no properties that would cause a prudent physician to avoid administering the substance to a patient, taking into account the disease or condition being treated and the individual route of administration. For example, such substances are generally required to be substantially sterile, for example, for injectables.

[0246] The term "carrier" refers, without limitation, to a flow promoter, diluent, adjuvant, excipient, or vehicle, etc., with which a compound is administered together. Examples of carriers are described herein and in Remington: The Science and Practice of Pharmacy (Remington: The Science and Practice of Pharmacy, 23rd Edition, ISBN-13: 978-0128200070).

[0247] The term "effective amount" or "therapeutically effective amount" refers to an amount sufficient to effect such treatment when administered to a mammal in need of the treatment as defined herein. The therapeutically effective amount will vary depending on the patient being treated, the weight and age of the patient, the severity of the disease state, the method of administration, etc., and can be readily determined by one of ordinary skill in the art.

[0248] The term "solvate" means a solid-state complex such as an adduct formed between an organic compound and solvent molecules. Solvates can be held together by hydrogen bonds, van der Waals forces, or other non-covalent interactions. Solvates can be channel solvates in that various amounts of solvent can be present in the channels of the solid structure.

[0249] The term "treatment" or "treating" includes, in the context of a disease or condition, preventing the occurrence of the disease or condition, inhibiting the disease or condition, eradicating the disease or condition, and / or alleviating one or more symptoms of the disease or condition.

[0250] Unless otherwise specifically stated, if a compound can exist as another tautomer, positional isomer and / or stereoisomer, all other isomers are intended to be included within the scope of the subject matter of this application. For example, if a compound is described as a particular optical isomer D- or L-, both optical isomers are intended to be included herein. For example, if a compound is described as having one of two tautomers, both tautomers are intended to be included herein. Thus, the compounds provided herein may be pure as enantiomers or may be mixtures of stereoisomers or diastereomers. The compounds provided herein may contain chiral centers. Such chiral centers may be either in the (R) or (S) configuration or mixtures thereof. The chiral centers of the compounds provided herein may undergo epimerization in vivo. Thus, those skilled in the art recognize that in the case of a compound that undergoes epimerization in vivo, administration of the (R) form of the compound is equivalent to administration of the (S) form of the compound.

[0251] This disclosure also includes all suitable isotopic variants of the compounds according to this disclosure, whether or not radioactive. Isotopic variants of the compounds according to this disclosure are understood to mean compounds in which at least one atom within the compounds according to this disclosure has been replaced with another atom having the same atomic number but a different atomic mass than the atomic mass that is normally or predominantly naturally occurring. Examples of isotopes that can be incorporated into the compounds according to this disclosure are 2 H (deuterium), 3 H (tritium), 13 C, 14 C, 15 N, 17 O, 18 O, 18 F, 36 Cl, 82 Br, 123 I, 124 I, 125 I, 129 I and 131Isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, chlorine, bromine and iodine, such as I. For example, in the case of testing the mechanism of action or the distribution of active compounds in the body, there may be advantages to certain isotope variants of the compounds according to the present disclosure, in particular compounds incorporating one or more radioisotopes. 3 H, 14 C and / or 18 Compounds labeled with F isotopes are suitable for this purpose. Furthermore, the incorporation of isotopes, such as deuterium, can result in a greater metabolic stability of the compound, leading to certain therapeutic benefits, such as an extended half-life in the body or a reduced dosage of the active substance required. In some embodiments, the hydrogen atoms of the compounds described herein may be replaced by deuterium atoms. In certain embodiments, unless otherwise indicated, "deuterated" as applied to a chemical group refers to a chemical group that is isotopically enriched with deuterium in an amount substantially greater than its natural abundance. Isotope variants of the compounds according to the present disclosure can be prepared, for example, by various means including the methods in the following and the examples, using the corresponding isotope modifications of specific reagents and / or starting compounds among them.

[0252] Accordingly, any of the embodiments described herein are intended to include single stereoisomers, mixtures of stereoisomers and / or isotopes of the compound.

[0253] Unless otherwise indicated, the terms “about” or “approximately” mean an acceptable error for a particular value as determined by one of ordinary skill in the art, which error is partly dependent on how the value is measured or determined. In certain embodiments, the terms “about” or “approximately” mean within 1, 2, or 3 standard deviations. In certain embodiments, the terms “about” or “approximately” mean within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.25%, 0.2%, 0.1%, or 0.05% of a given value or range. Unless otherwise specified, the term “about” means within plus or minus 10% of the explicitly recited value, rounded to the nearest integer and either rounded up or down.

[0254] Accordingly, the subject matter of the method is described by reference to certain embodiments discussed above. It is recognized that these embodiments are susceptible to various modifications and alternative forms that are well known to those of ordinary skill in the art.

[0255] The present disclosure may be further described by one or more of the following non-limiting clauses.

[0256] Clause 1. An amorphous calcium salt of obeticholic acid.

[0257] Clause 2. Amorphous obeticholic acid hemicalcium.

[0258] Clause 3. Stable amorphous obeticholic acid hemicalcium.

[0259] Clause 4. Substantially pure amorphous obeticholic acid hemicalcium.

[0260] Clause 5. An amorphous obeticholic acid hemicalcium salt substantially free of any crystalline salts of obeticholic acid hemicalcium, as described in Clauses 2-4.

[0261] Item 6. The amorphous obcetrapib hemicalcium of Items 2 to 5, having an X-ray powder diffraction pattern that is substantially the same as the X-ray powder diffraction pattern of FIG. 1.

[0262] Item 7. The amorphous obcetrapib hemicalcium of Items 2 to 5, having an X-ray powder diffraction pattern that includes one or more X-ray powder diffraction peaks at approximately 3.4° (2θ), approximately 7.0° (2θ), and approximately 9.2° (2θ).

[0263] Item 8. The amorphous obcetrapib hemicalcium of Items 2 to 7, wherein the amorphous obcetrapib hemicalcium does not exhibit birefringence.

[0264] Item 9. The amorphous obcetrapib hemicalcium of Items 2 to 8, having a glass transition temperature at a value between approximately 107°C and approximately 112°C.

[0265] Item 10. The amorphous obcetrapib hemicalcium of Item 9, wherein the glass transition temperature is measured by modulated differential scanning calorimetry.

[0266] Item 11. The amorphous obcetrapib hemicalcium of Item 10, wherein the measurement by modulated differential scanning calorimetry uses an open sample pan.

[0267] Item 12. The amorphous obcetrapib hemicalcium of Item 11, wherein the opening is a pinhole.

[0268] Item 13. The amorphous obcetrapib hemicalcium of Items 8 to 12, having a glass transition temperature at a value between approximately 110°C and approximately 112°C.

[0269] Item 14. The amorphous obcetrapib hemicalcium of Items 2 to 13, having a glass transition temperature of less than approximately 100°C when measured by differential scanning calorimetry using a sealed sample pan.

[0270] Clause 15. The amorphous obcetrapib hemicalcium of Clause 14, having a glass transition temperature at a value between about 70 °C and about 92 °C when measured by differential scanning calorimetry using a sealed sample pan.

[0271] Clause 16. The amorphous obcetrapib hemicalcium of Clauses 2 - 15, having a weight loss of less than about 1% when heated to about 200 °C.

[0272] Clause 17. The amorphous obcetrapib hemicalcium of Clause 16, having a weight loss between about 0.8% and about 0.95%.

[0273] Clause 18. The amorphous obcetrapib hemicalcium of Clause 17, having a weight loss between about 0.84% and about 0.92%.

[0274] Clause 19. The amorphous obcetrapib hemicalcium of Clauses 2 - 18, having a moisture content of less than about 5%.

[0275] Clause 20. The amorphous obcetrapib hemicalcium of Clause 19, having a moisture content of less than about 4%.

[0276] Clause 21. The amorphous obcetrapib hemicalcium of Clause 20, having a moisture content of less than about 3%.

[0277] Clause 22. The amorphous obcetrapib hemicalcium of Clause 19, having a moisture content between about 0.5% and about 1.5%.

[0278] Clause 23. The amorphous obcetrapib hemicalcium of Clauses 2 - 22, in bulk form or formulated composition having a particle size distribution in which about 90% of the particles have a diameter of about 15 microns or less.

[0279] Clause 24. The amorphous obcetrapib hemicalcium of Clause 23, in which about 90% of the particles have a diameter between about 6 microns and about 15 microns.

[0280] Clause 25. The amorphous obeticholic acid hemicalcium of Clause 24 having a particle size distribution in which about 90% or more of the particles have a diameter of about 14 microns or less.

[0281] Clause 26. The amorphous obeticholic acid hemicalcium of Clause 25 having a particle size distribution in which about 90% or more of the particles have a diameter of about 13 microns or less.

[0282] Clause 27. The amorphous obeticholic acid hemicalcium of Clause 26 having a particle size distribution in which about 90% or more of the particles have a diameter of about 12 microns or less.

[0283] Clause 28. The amorphous obeticholic acid hemicalcium of Clause 27 having a particle size distribution in which about 90% or more of the particles have a diameter of about 11 microns or less.

[0284] Clause 29. The amorphous obeticholic acid hemicalcium of Clause 28 having a particle size distribution in which about 90% or more of the particles have a diameter of about 10 microns or less.

[0285] Clause 30. The amorphous obeticholic acid hemicalcium of Clause 29 having a particle size distribution in which about 90% or more of the particles have a diameter of about 9 microns or less.

[0286] Clause 31. The amorphous obeticholic acid hemicalcium of Clause 30 having a particle size distribution in which about 90% or more of the particles have a diameter of about 8 microns or less.

[0287] Clause 32. The amorphous obeticholic acid hemicalcium of Clause 31 having a particle size distribution in which about 90% or more of the particles have a diameter of about 7 microns or less.

[0288] Clause 33. The amorphous obcetrapib hemicalcium of Clause 32, having a particle size distribution in which about 90% or more of the particles have a diameter of about 6 microns or less.

[0289] Clause 34. The amorphous obcetrapib hemicalcium of Clause 33, having a particle size distribution in which about 90% or more of the particles have a diameter of about 5 microns or less.

[0290] Clause 35. The amorphous obcetrapib hemicalcium of Clause 34, having a particle size distribution in which about 90% or more of the particles have a diameter of about 4 microns or less.

[0291] Clause 36. The amorphous obcetrapib hemicalcium of Clause 35, having a particle size distribution in which about 90% or more of the particles have a diameter of about 3 microns or less.

[0292] Clause 37. The amorphous obcetrapib hemicalcium of Clauses 2 - 36, in bulk form or in the form of a formulated composition, having a particle size distribution in which about 50% of the particles have a diameter of about 5 microns or less.

[0293] Clause 38. The amorphous obcetrapib hemicalcium of Clause 37, having a particle size distribution in which about 50% of the particles have a diameter of about 4 microns or less.

[0294] Clause 39. The amorphous obcetrapib hemicalcium of Clause 38, having a particle size distribution in which about 50% of the particles have a diameter of about 3 microns or less.

[0295] Clause 40. The amorphous obcetrapib hemicalcium of Clauses 2 - 39, in bulk form or in the form of a formulated composition, having a particle size distribution in which about 10% of the particles have a diameter of about 2 microns or less.

[0296] Clause 41. Amorphous obeticholic acid hemicalcium having a chemical purity of at least 98.0% and falling under Clauses 2 to 40.

[0297] Clause 42. Amorphous obeticholic acid hemicalcium having a chemical purity of at least 99.0% and falling under Clause 41.

[0298] Clause 43. Amorphous obeticholic acid hemicalcium having a chemical purity of at least 99.5% and falling under Clause 42.

[0299] Clause 44. Amorphous obeticholic acid hemicalcium having a chemical purity of at least 99.6% and falling under Clause 43.

[0300] Clause 45. Amorphous obeticholic acid hemicalcium having a chemical purity of at least 99.7% and falling under Clause 44.

[0301] Clause 46. Amorphous obeticholic acid hemicalcium having a chemical purity of at least 99.8% and falling under Clause 45.

[0302] Clause 47. Amorphous obeticholic acid hemicalcium having a chemical purity of at least 99.9% and falling under Clause 46.

[0303] Clause 48. The solid state of Figure 17 13 A solid state substantially the same as the 13 13 C-NMR spectrum and having an amorphous obeticholic acid hemicalcium falling under Clauses 2 to 47.

[0304] Clause 49. A solid state having a 13 13 C-NMR spectrum without a peak at approximately 22.1 ppm and having an amorphous obeticholic acid hemicalcium falling under Clauses 2 to 48.

[0305] Clause 50. A solid state having a 13 13 C-NMR spectrum without a peak at approximately 29.5 ppm and having an amorphous obeticholic acid hemicalcium falling under Clauses 2 to 49.

[0306] Clause 51. Unmilled amorphous obcetrapib hemicalcium.

[0307] Clause 52. Milled amorphous obcetrapib hemicalcium.

[0308] Clause 53. Amorphous obcetrapib hemicalcium according to Clauses 2 to 50, in which the amorphous obcetrapib hemicalcium has been milled.

[0309] Clause 54. Amorphous obcetrapib hemicalcium according to Clauses 2 to 50 or 53, in which the amorphous obcetrapib hemicalcium has been jet-milled.

[0310] Clause 55. Amorphous obcetrapib hemicalcium according to Clauses 2 to 50 or 53 to 54, in which the amorphous obcetrapib hemicalcium has been spray-dried.

[0311] Clause 56. Amorphous obcetrapib hemicalcium prepared by a synthetic method, in which the intermediate in said method comprises crystalline obcetrapib HCl.

[0312] Clause 57. Amorphous obcetrapib hemicalcium according to Clauses 2 to 56, prepared by a synthetic method, in which the intermediate in said method comprises crystalline obcetrapib HCl.

[0313] Clause 58. Obcetrapib HCl.

[0314] Clause 59. Crystalline obcetrapib HCl.

[0315] Clause 60. Amorphous obcetrapib HCl compound.

[0316] Clause 61. Solvates of obcetrapib HCl according to Clauses 58 to 60.

[0317] Clause 62. Obeticholic acid HCl of Clauses 58 - 61, wherein the weight percentage of HCl is between about 0.01% and about 8%.

[0318] Clause 63. A composition comprising any one of the crystalline obeticholic acid HCl of Clauses 58 - 62.

[0319] Clause 64. The crystalline obeticholic acid HCl of Clauses 58 - 60 or 62 - 63, wherein the crystalline obeticholic acid HCl is a solvate.

[0320] Clause 65. The crystalline obeticholic acid HCl of Clause 64, wherein the solvate comprises obeticholic acid and hydrochloric acid.

[0321] Clause 66. The crystalline obeticholic acid HCl of Clause 65, wherein the solvate comprises an organic solvent.

[0322] Clause 67. The crystalline obeticholic acid HCl of Clause 66, wherein the solvate comprises a solvent having sufficient solubility to dissolve sufficient HCl to provide sufficient HCl to form crystalline obeticholic acid HCl.

[0323] Clause 68. The solvate of any one of Clauses 61 or 64 - 67, wherein the solvent of the solvate is selected from methanol, ethanol, isopropanol, acetic acid, acetonitrile, acetone, methyl isobutyl ketone, isopropyl acetate, tetrahydrofuran, methyl t-butyl ether, cyclopentyl methyl ether (CPME), N-methyl-2-pyrrolidone, dimethyl sulfoxide, dimethylformamide, 2-methyl-tetrahydrofuran, dichloromethane, 1,4-dioxane, 1,2-difluorobenzene, toluene, and hexafluoroisopropanol.

[0324] Clause 69. The crystalline obeticholic acid HCl of Clause 68, wherein the solvent is CPME.

[0325] Clause 70. Crystalline obcetrapib HCl of any one of Clauses 58-59 or 61-69 having an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern in FIG. 19.

[0326] Clause 71. Crystalline obcetrapib HCl of any one of Clauses 58-59 or 61-69 having an X-ray powder diffraction pattern containing a peak at about 9.8° (2θ).

[0327] Clause 72. Crystalline obcetrapib HCl of any one of Clauses 58-59, 61-69 or 71 having an X-ray powder diffraction pattern containing one or more peaks at about 8.1° (2θ), about 9.8° (2θ), about 13.8° (2θ), about 16.7° (2θ) and about 19.5° (2θ).

[0328] Clause 73. A salt according to formula (VI):

Chemical formula

[0329] Clause 74. A salt according to Clause 73, wherein the compound is the mesylate salt (Compound 1D) of the following structure:

Chemical formula

[0330] Clause 75. The crystalline mesylate salt of Compound 1D of Clause 74.

[0331] Clause 76. The crystalline mesylate salt of Compound 1D of Clause 75 having a powder diffraction pattern substantially the same as any one of the four X-ray powder patterns described in FIG. 20.

[0332] Clause 77. The crystalline mesylate salt of Compound 1D of Clause 75 having an X-ray powder diffraction pattern containing one or more peaks at about 5.2° (2θ) and about 9.1° (2θ).

[0333] Clause 78. A crystalline mesylate of Compound 1D of Clauses 75 - 77 having an X-ray powder diffraction pattern comprising one or more peaks at about 9.1° (2θ), about 15.9° (2θ), about 16.5° (2θ), about 17.2° (2θ), about 18.6° (2θ) and about 19.2° (2θ).

[0334] Clause 79. A method for preparing obicetrapib, comprising: (a) preparing a compound of formula (IV) by coupling a compound of formula (II) or a salt thereof with a compound of formula (III);

Chemical formula

Chemical formula

Chemical formula

[0335] ​Clause 80. The compound of formula (II) in step (a) is, prior to step (a), as follows: (Pre-a1) A compound of formula (IIA) or (IIB): [Chemical formula] The step of preparing, and (Pre-a2) The step of degrading the compound of formula (IIA) or (IIB) by salt decomposition to obtain the compound of formula (II) A method according to clause 79, obtained by applying, The reaction in step (Pre-a2) is carried out in an organic solvent, the compound of formula (II) is not isolated from the organic solvent depending on the situation, and the method does not require chromatography.

[0336] Clause 81. The salt of formula (IIA) or (IIB) is selected from salts with anions Am− selected from sulfonate ion, sulfate ion, halogen, acetate ion, aspartate ion, benzoate ion, bicarbonate ion, bitartrate ion, carbonate ion, citrate ion, decanoate ion, fumarate ion, gluceptate ion, gluconate ion, glutamate ion, glycolate ion, hexanoate ion, hydroxynaphthoate ion, isethionate ion, lactate ion, lactobionate ion, malate ion, maleate ion, mandelate ion, mucate ion, nitrate ion, octanoate ion, oleate ion, pamoate ion, pantothenate ion, phosphate ion, polygalacturonate ion, propionate ion, salicylate ion, stearate ion, succinate ion, tartrate ion and theophyllinate ion, the sulfonate ion may be besylate ion, tosylate ion, napsylate ion, camsylate ion, esylate ion, edisylate ion or mesylate ion, the sulfate ion may be methylsulfate ion, and the halogen may be chloride ion, iodide ion or bromide ion, a method according to clause 80.

[0337] Clause 82. Anion A m-The method of clause 81, wherein the salt with [substance] is selected from chloride, bromide, bitartrate, sulfate and sulfonate.

[0338] Clause 83. Anion A m- The method of clause 82, wherein the salt with [substance] is selected from chloride, bromide, bitartrate and mesylate.

[0339] Clause 84. Y in the compounds of formulas (III) to (VI) and (VIII) 1 The method of any one of clauses 79 to 83, wherein [substance] is selected from an alkyl group, a substituted alkyl group, an aryl group, a substituted aryl group, an allyl group, a substituted allyl group and a silyl group.

[0340] Clause 85. Y in the compounds of formulas (III) to (VI) and (VIII) 1 The method of clause 84, wherein [substance] is selected from t-butyl, methyl, ethyl, benzyl, allyl, substituted allyl, 2,2,2-trifluoroethyl, phenyl, 4-methoxybenzyl ester, 2,6-disubstituted phenol and silyl group.

[0341] Clause 86. Y in the compounds of formulas (III) to (VI) and (VIII) 1 The method of clause 85, wherein [substance] is t-butyl.

[0342] Clause 87. The salt of formula (VI) is selected from anions A which are sulfonate ion, sulfate ion, halogen, acetate ion, aspartate ion, benzoate ion, bicarbonate ion, bitartrate ion, carbonate ion, citrate ion, decanoate ion, fumarate ion, gluceptate ion, gluconate ion, glutamate ion, glycolate ion, hexanoate ion, hydroxynaphthoate ion, isethionate ion, lactate ion, lactobionate ion, malate ion, maleate ion, mandelate ion, mucate ion, nitrate ion, octanoate ion, oleate ion, pamoate ion, pantothenate ion, phosphate ion, polygalacturonate ion, propionate ion, salicylate ion, stearate ion, succinate ion, tartrate ion and theophylline ion. n- selected from salts with, wherein the sulfonate ion may be besylate ion, tosylate ion, napsylate ion, camsylate ion, esylate ion, edisylate ion or mesylate ion, the sulfate ion may be methyl sulfate ion, and the halogen may be chloride ion, iodide ion or bromide ion, according to any one of the methods of Clauses 79 to 86.

[0343] Clause 88. The salt with anion A n- is selected from chloride, bromide, bitartrate, sulfate and sulfonate, according to the method of Clause 87.

[0344] Clause 89. The salt with anion A n- is selected from chloride, bromide, bitartrate and mesylate, according to the method of Clause 87.

[0345] Clause 90. The salt form of formula (VI) is the mesylate of Compound 1D:

Chemical formula

[0346] Clause 91. The method of Clause 90, wherein the mesylate is crystalline.

[0347] Clause 92. X in the compound of formula (III) 1 is selected from halogen, carbamate and substituted sulfonyloxy groups, any one of the methods of Clauses 79 to 91.

[0348] Clause 93. X in the compound of formula (III) 1 is halogen, the method of Clause 92.

[0349] Clause 94. The method of Clause 93, wherein the halogen is chloride.

[0350] Clause 95. X in the compound of formula (VII) 2 is selected from halogen and substituted sulfonyloxy groups, any one of the methods of Clauses 79 to 94.

[0351] Clause 96. X in the compound of formula (III) 2 is halogen, the method of Clause 95.

[0352] Clause 97. The method of Clause 96, wherein the halogen is bromide.

[0353] Clause 98. A method for preparing an amorphous hemicalcium salt of obicetrapib, comprising (i) treating obicetrapib with HCl to obtain a crystalline obicetrapib HCl compound, (ii) isolating the crystalline obicetrapib HCl compound, (iii) preparing an amorphous hemicalcium salt of obicetrapib from the crystalline obicetrapib HCl compound isolated in step (ii), and (iv) isolating the amorphous hemicalcium salt of obicetrapib A method comprising.

[0354] Clause 99. The crystalline obicetrapib HCl compound isolated in step (ii) is a compound of formula (IH):

Chemical formula

[0355] Clause 100. The preparation of the amorphous hemicalcium salt of formula (I) in step (iii) is as follows: (iii-1) Converting the crystalline obicetrapib HCl compound of step (ii) in one or more suitable solvents selected from organic solvents and aqueous solvents to obtain obicetrapib; (iii-2) Treating obicetrapib in an organic solvent with aqueous sodium hydroxide to form the sodium salt of obicetrapib; and (iii-3) Treating the sodium salt of obicetrapib with aqueous calcium chloride to form the amorphous hemicalcium salt of obicetrapib The method according to clause 98 or 99, comprising The compounds in steps (iii-1) and (iii-2) are not isolated depending on the circumstances.

[0356] Clause 101. The method according to any one of clauses 98 to 100, wherein the amorphous hemicalcium salt of obicetrapib is amorphous obicetrapib hemicalcium.

[0357] Clause 102. The method according to any one of clauses 98 to 101, wherein the amorphous calcium salt of obicetrapib is isolated with a chemical purity of at least 99%.

[0358] Clause 103. The method of clause 102, wherein the amorphous calcium salt of obicetrapib is isolated with a purity of at least 99.1%.

[0359] Clause 104. The method of clause 102, wherein the amorphous calcium salt of obicetrapib is isolated with a purity of at least 99.2%.

[0360] Article 105. The method of Article 102, wherein the amorphous calcium salt of obeticholic acid is isolated with a purity of at least 99.3%.

[0361] Article 106. The method of Article 102, wherein the amorphous calcium salt of obeticholic acid is isolated with a purity of at least 99.4%.

[0362] Article 107. The method of Article 102, wherein the amorphous calcium salt of obeticholic acid is isolated with a purity of at least 99.5%.

[0363] Article 108. The method of Article 102, wherein the amorphous calcium salt of obeticholic acid is isolated with a purity of at least 99.6%.

[0364] Article 109. The method of Article 102, wherein the amorphous calcium salt of obeticholic acid is isolated with a purity of at least 99.7%.

[0365] Article 110. The method of Article 102, wherein the amorphous calcium salt of obeticholic acid is isolated with a purity of at least 99.8%.

[0366] Article 111. The method of Article 102, wherein the amorphous calcium salt of obeticholic acid is isolated with a purity of at least 99.9%.

[0367] Article 112. The method according to any one of Articles 102 to 111, wherein the amorphous calcium salt of obeticholic acid is amorphous obeticholic acid hemicalcium.

[0368] Article 113. A pharmaceutical composition comprising an amorphous salt of obeticholic acid calcium according to any one of Articles 1 to 57 and one or more pharmaceutically acceptable carriers.

[0369] Article 114. The pharmaceutical composition of Article 113, wherein the amorphous salt of obeticholic acid calcium is amorphous obeticholic acid hemicalcium.

[0370] Article 115. A method for treating a subject suffering from a cardiovascular disease or having an increased risk of developing the same, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition according to Article 113 or 114.

[0371] Article 116. An amorphous calcium salt of obeticholic acid prepared by any one of the methods of Articles 79 to 112.

[0372] Article 117. The amorphous calcium salt of Article 116, which is amorphous obeticholic acid hemicalcium.

[0373] Article 118. A method for preparing an amorphous obeticholic acid calcium salt, the method comprising treating obeticholic acid with an acid to form a salt, solvate, composition or combination thereof; isolating the salt, solvate, composition or combination thereof; and treating the salt, solvate, composition or combination thereof with a calcium source to prepare an amorphous obeticholic acid hemicalcium salt.

[0374] Article 119. The method of Article 118, wherein the calcium source is calcium chloride.

[0375] Article 120. A salt, solvate, composition or combination thereof comprising obeticholic acid and a free acid.

[0376] Article 121. The salt of Article 120.

[0377] Article 122. The solvate of Article 120.

[0378] Article 123. The composition of Article 120.

[0379] Clause 124. A salt, solvate, composition, or combination thereof according to Clause 120, wherein the free acid is selected from sulfonic acid, sulfuric acid, halogenated acid, acetic acid, aspartic acid, benzoic acid, bicarbonate, bitartrate, carbonic acid, citric acid, decanoic acid, fumaric acid, gluceptic acid, gluconic acid, glutamic acid, glycolic acid, hexanoic acid, hydroxynaphthoic acid, isethionic acid, lactic acid, lactobionic acid, malic acid, maleic acid, mandelic acid, mucic acid, nitric acid, octanoic acid, oleic acid, pamoic acid, pantothenic acid, phosphoric acid, polygalacturonic acid, propionic acid, salicylic acid, stearic acid, succinic acid, tartric acid, and theocluic acid; the sulfonic acid may be benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, ethanedisulfonic acid, or methanesulfonic acid; the sulfuric acid may be methylsulfuric acid; and the halogenated acid may be HCl, HBr, or HI.

[0380] Clause 125. The method according to Clause 118, wherein the calcium source is a calcium halide salt.

[0381] Clause 126. The method according to Clause 118, wherein the calcium source is a soluble calcium salt.

[0382] Clause 127. The method according to Clause 118, wherein the calcium source is a calcium salt.

[0383] Clause 128. The obeticholic acid hydrochloride according to Clause 58 or 59, which contains a chloride ion hydrogen-bonded to at least one protonated nitrogen on the pyrimidine ring of obeticholic acid.

[0384] Clause 129. The obeticholic acid hydrochloride according to Clause 59 or 128, which has an asymmetric unit containing four cationic obeticholic acid moieties, two neutral obeticholic acid molecules, four chloride anions, and at least one solvent molecule.

[0385] Clause 130. The obeticholic acid hydrochloride according to Clause 129, wherein the asymmetric unit contains two solvent molecules.

[0386] Clause 131. The obeticholic acid hydrochloride of Clause 129, wherein the asymmetric unit contains three solvent molecules.

[0387] Clause 132. The obeticholic acid hydrochloride according to any one of Clauses 129 to 132, wherein the solvent molecule is selected from heptane and cyclopentyl methyl ether.

[0388] Clause 133. The following unit cell parameters: [Table 14] The obeticholic acid hydrochloride according to any one of Clauses 128 to 132, having:

[0389] Clause 134. The crystalline obeticholic acid hydrochloride of Form A.

[0390] Clause 135. The crystalline obeticholic acid hydrochloride of Form A according to Clause 134, having an X-ray powder diffraction pattern including a peak at about 8.6° (2θ), two peaks between about 9.7° (2θ) and about 10.4° (2θ), and two peaks between about 8.6° (2θ) and 9.0° (2θ).

[0391] Clause 136. The crystalline obeticholic acid hydrochloride of Form A according to Clause 134, having an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern of Figure 22.

[0392] Clause 137. The crystalline obeticholic acid hydrochloride of Form B.

[0393] Clause 138. The crystalline obeticholic acid hydrochloride of Form B according to Clause 136, having an X-ray powder diffraction pattern including peaks at about 6.5° (2θ), about 8.8° (2θ), and about 11.0° (2θ).

[0394] Clause 139. The crystalline obeticholic acid hydrochloride of Form B according to Clause 137, having an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern of Figure 23.

[0395] Clause 140. Crystalline obeticholic acid hydrochloride in Form C.

[0396] Clause 141. Crystalline obeticholic acid hydrochloride in Form C of Clause 140, having an X-ray powder diffraction pattern that is substantially the same as the X-ray powder diffraction pattern of FIG. 24.

[0397] Clause 142. Crystalline obeticholic acid hydrochloride in Form D.

[0398] Clause 143. Crystalline obeticholic acid hydrochloride in Form D of Clause 142, having an X-ray powder diffraction pattern that is substantially the same as the X-ray powder diffraction pattern of FIG. 25.

[0399] Clause 144. Crystalline obeticholic acid hydrochloride of Clause 59, having an X-ray powder diffraction pattern that includes a peak between approximately 4.3° (2θ) and approximately 4.7° (2θ).

Examples

[0400] The examples in this section are presented by way of illustration and not limitation. It should be understood that the examples can only represent some embodiments, and the following examples are illustrative and not limiting. All substituents are as defined heretofore unless otherwise specified. Reagents and starting materials are readily available to those skilled in the art. The specific synthetic steps for each of the described routes can be combined in various ways or coordinated with steps from different schemes to prepare the compounds described herein.

Chemical Formula

[0401] Referring to Scheme 1, amorphous obeticholic acid calcium (Compound 3) was prepared in six chemical steps and three isolations from (2R,4S)-4-amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline mesylate (Compound 1A), t-butyl-4-(2-chloropyrimidin-5-yloxy)-butyrate (Compound 1B), and 3,5-bis(trifluoromethyl)benzyl bromide (Compound 1E). Compound 1A was coupled with Compound 1B by a palladium-catalyzed reaction to produce a solution of (2R,4S)-4-[5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl)]amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline (Compound 1C), which was not isolated and was reacted directly with an excess of ethyl chloroformate in the presence of pyridine to produce ethyl (2R,4S)-4-[5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl)]amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate, which was isolated as the crystalline mesylate (Compound 1D). Alkylation of the crystalline mesylate, Compound 1D, with 3,5-bis(trifluoromethyl)benzyl bromide (Compound 1E) under strongly basic conditions produced a toluene solution of ethyl (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate (Compound 1F). Next, treatment of Compound 1F with acid cleavage of the tert-butyl ester produced a solution of ethyl (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate (Compound 1).Next, compound 1 was converted to compound 2, which is a solvate of ethyl (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate (compound 2). Finally, compound 2 was converted to an amorphous hemicalcium salt (compound 3) and milled to the target particle size. Compound 2 is crystalline obicetrapib HCl, and compound 3 is amorphous obicetrapib hemicalcium. The FT-IR spectrum of the milled amorphous obicetrapib hemicalcium can be seen in Figure 4. In the solution state corresponding to the chemical structure of obicetrapib hemicalcium. 1 The 1H-NMR spectrum can be seen in Figure 5.

[0402] Each step in the production process of ethyl (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate (compound 1), intermediate (compound 2) which is intermediate HCl, and the corresponding amorphous calcium salt (compound 3) will be described in more detail in the following Examples 1 to 16.

[0403] Examples 1 to 3, 5, 7, 9, 11 to 12 describe the method of production steps in the method for preparing amorphous obicetrapib hemicalcium (compound 3), and Examples 4, 6, 8, 10 and 13 present additional methods for preparing the indicated compounds. The methods in these examples sometimes represent more than one batch of the prepared indicated compound.

[0404] Examples 14 to 15 describe the method of milling amorphous obicetrapib hemicalcium (compound 3), and Example 16 describes the method of preparing crystalline obicetrapib hemicalcium. (Example 1) (2R,4S)-4-Amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline (free base of Compound 1A) Preparation

Chem.

[0405] Into a reaction vessel equipped with a reflux condenser, (2R,4S)-4-amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline (Compound 1A) (62 kg, 182 mol, 1.00 equivalent) was added together with toluene (375 L). The resulting slurry was stirred at 52 °C, and 1 M aqueous sodium hydroxide solution (322 L, 5.2 volumes) was added. The reaction mixture was stirred until all solids dissolved, and then cooled to 20 °C. Stirring was stopped, and the reaction mixture was separated into two phases. The lower aqueous phase was drained, and an aqueous sodium chloride solution (310 L, 5.0 volumes) was added. Next, the reaction mixture was stirred at 20 °C for 30 minutes. Stirring was stopped again, and the reaction mixture was separated into two phases. The lower aqueous phase was drained, and deionized water (310 L, 5.0 volumes) was added. Next, the reaction mixture was stirred at 20 °C for 30 minutes. Stirring was stopped again, and the reaction mixture was separated into two phases. The lower aqueous phase was separated. Next, the resulting organic solution was distilled under vacuum at an internal temperature of 65 °C or less. Distillation was continued until a final visual volume of 4.0 volumes (250 L) was reached. Next, the reaction vessel was cooled to 20 °C to obtain a toluene solution of (2R,4S)-4-amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline (Compound 1A-free base) containing a small amount of water. Compound 1A-free base was not isolated and was used directly in Example 2. (Example 2) (2R,4S)-4-[5-(3-t-Butoxycarbonylpropoxy)pyrimidin-2-yl)]amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline (Compound 1C) Preparation

Chem.

[0406] To a reaction vessel (“Vessel A”) containing (2R,4S)-4-amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline (Compound 1A - free base) in toluene containing <1000 ppm of water derived from the previous step, additional toluene (107 L, 1.5 volumes) was added. Next, to Vessel A, t-butyl-4-(2-chloropyrimidin-5-yloxy)-butyrate (Compound 1B) (54.6 kg, 200 mol, 1.10 equivalents) was added together with t-BuOH (122 L, 1.55 volumes). The reaction mixture was stirred and nitrogen was bubbled through. During this time, to a second reaction vessel (“Vessel B”), palladium acetate (410 g, 1.8 mol, 1 mol%) was added under nitrogen. To Vessel B, (S)-BINAP (2.48 kg, 4.0 mol, 2.2 mol%) and toluene (107 L, 1.5 volumes) were further added, and when the resulting mixture was stirred, a red / orange Pd-BINAP solution was formed. The orange / red Pd-BINAP solution in reaction vessel B was transferred to Vessel A. To Vessel A, K3PO4 (85 kg, 400 mol, 2.20 equivalents) was further added, and the resulting reaction mixture was heated to an internal temperature of 72 °C and stirred for at least 2 hours. Next, the mixture was cooled to 20 °C, deionized water (124 L) was carefully added, and the mixture was stirred for 30 minutes. Next, stirring was stopped and the layers were separated into two phases. The lower aqueous phase was separated, and an aqueous solution of 1M HCl (123 L) was added with stirring. After 30 minutes, stirring was stopped again and the layers were separated into two phases. The lower aqueous phase was separated, and an aqueous solution of sodium chloride (326 kg, 5.26 volumes) was added with stirring. After 30 minutes, stirring was stopped again and the layers were separated into two phases. The lower aqueous phase was separated, and deionized water (248 L, 4.0 volumes) was added with stirring. After 30 minutes, stirring was stopped again and the layers were separated into two phases. The lower aqueous phase was separated. Next, the resulting reaction mixture was treated with ethylenediamine (1.60 kg, 0.15 equivalents) and stirred at 20 °C for 80 minutes. Next, the reaction mixture was filtered through a cartridge of charcoal and the filtrate was returned to a clean vessel. Next, the mixture was distilled under partial vacuum at an internal temperature of 60 °C or less. Distillation was continued by visual observation in the reactor until approximately 2.50 volumes (155 L) remained, and then acetonitrile (394 L, 5.0 volumes) was added.Next, the mixture was distilled under vacuum at an internal temperature of 60 °C or less. Distillation was continued by visual observation in the reactor until it reached approximately 2.50 volumes (155 L), and then the contents were cooled to 20 °C. Next, acetonitrile (394 L, 5.0 volumes, until reaching 11 volumes by visual observation (approximately 620 L)) was added to the reaction vessel and (2R,4S)-4-[5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl)]amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline (Compound 1C) dissolved in acetonitrile was obtained. Compound 1C was not isolated and was used directly in Example 3. (Example 3) Preparation of Ethyl (2R,4S)-4-[5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl)]amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate as the Crystalline Mesylate (Compound 1D) [Chemical Formula]

[0407] (2R,4S)-4-[5-(3-t-Butoxycarbonylpropoxy)pyrimidin-2-yl)]amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline (Compound 1C) in acetonitrile (approx. 620 L) was cooled to an internal temperature of <10 °C, and pyridine (72 L, 900 mol, 4.9 equivalents) was added. Next, while maintaining the internal temperature of the reactor contents at <10 °C, ethyl chloroformate (136 L, 1428 mol, 7.84 equivalents) was added from a dropping funnel. Next, the internal temperature of the reaction mixture was linearly increased to 20 °C over 3.5 hours. Next, this mixture was distilled under vacuum at an internal temperature of 60 °C or less. Distillation was continued by visual observation until it reached approximately 2.50 volumes (155 L). Next, isopropyl acetate (471 L, 6.6 volumes) was added to the reaction vessel, and distillation was continued under vacuum at an internal temperature of 60 °C or less until approximately 2.50 volumes (155 L) remained by visual observation. Next, isopropyl acetate (471 L, 6.6 volumes), 1 M hydrochloric acid (307 L, 5.0 volumes), and 26% aqueous sodium chloride (63 L, 1.2 volumes) were added to the reaction vessel. The resulting mixture was stirred for 30 minutes and then separated into two phases. The lower aqueous phase was separated, and saturated aqueous sodium bicarbonate (132 L, 2.3 volumes) was added. The resulting mixture was stirred for 30 minutes and then separated into two phases. The lower aqueous phase was separated, and the residual mixture was distilled under vacuum and at 60 °C or less by visual observation until the total volume reached approximately 4.0 volumes (250 L), and ethyl (2R,4S)-4-[5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl)]amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate (the corresponding free base of Compound 1D) in isopropyl acetate was obtained with respect to the weight of the solution.

[0408] Additional isopropyl acetate (86 L, 1.4 volumes) and methyl t-butyl ether (MTBE, 593 L, 9.6 volumes) were added to the corresponding free base of Compound 1D in isopropyl acetate, and the jacket temperature was set to 20 °C. Next, methanesulfonic acid (MsOH, 17.6 kg, 1.0 equivalent relative to the mmol of the compound (the corresponding free base of Compound 1D)) was added to this reaction mixture over 60 minutes. Next, the resulting slurry was stirred for 8 hours. Next, the slurry was filtered under vacuum at 20 °C. Next, the solid cake was washed with a 75 / 25 v / v solution of isopropyl acetate (78 L, 1.1 volumes) and methyl t-butyl ether (236 L, 2.8 volumes), and then dried under vacuum at 20 °C, and the isolated (2R,4S)-4-[5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl)]amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester was obtained as a crystalline mesylate (Compound 1D) in a 74% yield relative to the number of moles of Compound 1A. The purity of the obtained crystalline Compound 1D was >99%. (Example 4) Further Preparation of Compounds 1C and 1D

[0409] Multiple batches of Compounds 1C and 1D were prepared generally using the preparations described below. In some preparations, for example, as further discussed below, seeding with Compound 1D was performed, and in others it was not.

[0410] Pd(OAc)2 and (S)-BINAP were dissolved in toluene and stirred to form the corresponding Pd-BINAP-complex (color change to red) (the "catalyst solution"). Toluene, Compound 1B, Compound 1A, and K3PO4 were added to the reactor and stirred. The target water content was about 6%. The catalyst solution was added to the reactor mixture, and the reaction mixture was heated to 70 - 75 °C and stirred.

[0411] Washed with HCl, brine, and water, followed by phase separation, then EDA and toluene were added, and this solution was stirred for approximately 90 minutes. To remove palladium, the solution was passed through a cartridge loaded with activated carbon (Begerow, F-9120). Thereafter, the solvent was exchanged from toluene to acetonitrile (MeCN) by distillation to obtain Compound 1C.

[0412] Pyridine was added to the solution of Compound 1C and cooled to below 10 °C before the addition of ethyl chloroformate. While controlling the temperature at NMT of 10 °C, ethyl chloroformate was administered to the solution of Compound 1C in one or two portions. Next, this reaction mixture was stirred at 17 - 27 °C for approximately 1 hour to convert Compound 1C to the free base of Compound 1D (Compound 1D-FB). Solvent exchange from acetonitrile to isopropyl acetate (iPrOAc) was carried out by distillation, and the organic phase was washed with HCl (1M), brine, and aqueous NaHCO3 (NaOH may be used), and then the volume was reduced by distillation.

[0413] Methanesulfonic acid (MsOH) and MTBE were added to the iPrOAc solution of Compound 1D-FB and stirred. In some cases, seed crystals of Compound 1D prepared previously were added, but it was not necessary. Whether or not seeds were added, crystallization of Compound 1D occurred thereafter. The solid product was filtered, washed with MTBE / iPrOAc (75 / 25), and dried on the filter. (Example 5) (2R,4S)-4-{[3,5-Bis(trifluoromethyl)benzyl]-[5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (Compound 1F) Preparation [Chemical formula]

[0414] In a reaction vessel at a temperature of 5 °C, ethyl (2R,4S)-4-[5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl]amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate was used as the crystalline mesylate (Compound 1D) (42 kg), and toluene (465 kg, 12.7 volumes) was added. Next, tetrabutylammonium hydrogen sulfate (3.5 kg, 0.16 equivalent) and sodium tert-pentoxide (34.5 kg, 4.8 equivalents) were added, and the resulting reaction mixture was stirred for 10 minutes and degassed with nitrogen. Next, 3,5-bis(trifluoromethyl)benzyl bromide (Compound 1E) (28 kg, 1.41 equivalents) was added to this reaction mixture, and stirring was continued at 5 °C for 6.5 hours. Next, this reaction mixture was treated with a 1N acetic acid solution (320 kg) and stirred at 20 °C for approximately 30 minutes. After this time, stirring was stopped and the mixture was separated into two phases. The lower aqueous phase was discarded, and the reaction mixture was concentrated under vacuum at an internal temperature of 60 °C or less until approximately 3.3 volumes (137 L) remained, and a toluene solution of 36.8 wt% of (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate (Compound 1F) was obtained in a yield of 97% based on the number of moles of Compound 1D. (Example 6) Further Preparation of Compound 1F

[0415] Compound 1E was added to a toluene solution containing Compound 1D and tetrabutylammonium hydrogen sulfate. Sodium tert-pentoxide in toluene was added under cooling. The resulting reaction mixture was quenched with dilute acetic acid. The aqueous layer was separated, the product in the toluene layer was treated with charcoal, and concentrated in vacuo (Compound 1F). (Example 7) (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (Compound 1) [Chemical formula]

[0416] A toluene solution of 37% by weight of ethyl (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl][5-(3-t-butoxycarbonylpropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate (Compound 1F) (128.4 kg of a 37% by weight solution, equivalent to 47.5 kg of Compound 1F) was diluted to 32% by weight with additional toluene, and then mixed with acetic acid (253 kg, 5.33% by weight) and 6M HCl (109.9 kg, 2.32% by weight, prepared in situ using 66.1 kg of concentrated HCl and 43.8 kg of water). The resulting reaction mixture was stirred vigorously and warmed to 48 °C for 3 hours. Next, the reaction mixture was cooled to 21 °C, and then n-heptane (159.8 kg, 3.36% by weight), acetonitrile (73.8 kg, 1.55% by weight), and water (170 kg, 3.58% by weight) were added. The resulting mixture was stirred for 34 minutes and then separated into two phases. Next, the lower aqueous phase was further treated with water (90 kg, 1.89% by weight), n-heptane (95 kg, 2.00% by weight), acetonitrile (38 kg, 0.80% by weight), and toluene (42 kg, 0.88% by weight), stirred again for 20 minutes, and then the organic phase was separated and the lower aqueous phase was discharged. Next, the combined organic phases were treated with water (240 kg, 5.05% by weight), stirred for an additional 30 minutes, and then separated into two phases. The lower aqueous phase was discarded, and the upper organic phase was treated with 5% w / w trisodium citrate dihydrate (34 kg, 0.72% by weight) and water (205 kg, 4.32% by weight). The resulting mixture was stirred vigorously for 30 minutes and then separated into two phases, after which the lower aqueous phase was discarded. The residual organic phase was treated again with water (240 kg, 5.05% by weight), stirred for 30 minutes, and then the two phases were separated and the lower aqueous phase was discharged. Next, while maintaining an internal temperature of 50 °C or less, the organic phase was concentrated in vacuo to approximately 3 volumes (approximately 149 L). The reaction mixture was diluted with cyclopentyl methyl ether (CPME, 250 kg, 5.26% by weight) and stirred. Next, this solution was concentrated in vacuo to approximately 3 volumes (approximately 165 L) while maintaining an internal temperature of 50 °C or less.Next, CPME (250 kg, 5.26 wt%) was added, and the mixture was concentrated in vacuo to approximately 2.5 volumes (approximately 124 L) while maintaining an internal temperature of 50 °C or less, to obtain a cyclopentyl methyl ether (CMPE) solution of 33.7 wt% of ethyl (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate (Compound 1, free base form) having 1 wt% of toluene and less than 1 wt% of n-heptane with respect to the weight of the solution. (Example 8) Further Preparation of Compound 1

[0417] Compound 1F (in solution in toluene) was mixed with acetic acid and 6 M aqueous HCl. The biphasic mixture was stirred vigorously at 45 - 50 °C and then cooled to 20 °C. After the addition of water, acetonitrile and n-heptane, the mixture was extracted and the layers were separated.

[0418] The aqueous layer of the first extraction was diluted with water and extracted a second time with acetonitrile, n-heptane and toluene. The two organic extracts obtained were combined. The organic phase was washed with water, 5% sodium citrate solution was added to bring the pH to ≧ 3.5. Washing with water was carried out and the organic layer was treated with activated carbon. Solvent exchange from toluene and n-heptane to CPME was carried out by repeating vacuum distillation and the addition of CPME to obtain Compound 1. (Example 9) (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate hydrochloride (Compound 2)

Chemical formula

[0419] A solution of 33.7 wt% ethyl (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl][5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate (Compound 1, free base form, 115.6 kg, 59.2 mol) derived from the previous step in cyclopentyl methyl ether (CPME) was added to a clean reaction vessel under nitrogen at a jacket temperature of 22 °C. After dilution with CPME (27.8 kg / 0.58 wt), when n-heptane (54.8 kg, 1.15 wt) was added, the internal reaction temperature rose to 39 °C. Next, while maintaining the internal reaction temperature at 39 °C, 3.0 M HCl (17.6 kg, 0.37 wt) in CPME was added at a constant rate. After completion of the addition of HCl, the internal temperature was raised to 52 °C. Next, while maintaining the internal reaction temperature at 51 °C, additional n-heptane (133.2 kg, 2.80 wt) was added at a constant rate. The reaction mixture was heated to 55 °C and then cooled to 49 °C. A certain portion of the reaction mixture was taken out and cooled to 11 °C at a linear cooling rate until a slurry containing crystals of Compound 2 (referred to herein as the "seed crystal slurry") was formed in CPME / n-heptane. Next, at 49 °C, a seed crystal slurry of Compound 2 (169 g, 0.43 wt%) in CPME / n-heptane was added and this temperature was held for 105 minutes. Next, the opaque reaction mixture was cooled to 11 °C at a linear cooling rate over a period of 12 hours. Next, the reaction mixture was filtered at 11 °C under vacuum and the solid wet HCl intermediate (Compound 2) was collected. Next, a mixture of CPME and n-heptane (56.6 kg of CPME, 179 kg of n-heptane) was added to the reaction vessel and cooled to 11 °C. Next, half of the mixture was poured through a filter dryer as a chromatography wash solution. The latter half was passed through the filter as a slurry wash solution. Without removing Compound 2 from the filter dryer, it was further purified by recrystallization according to the following procedure.

[0420] To a filter dryer containing Compound 2, Compound 2 in cyclopentyl methyl ether (CPME) (77.6 kg) was added and heated to 25 °C. Next, the dissolved Compound 2 was transferred to a reaction vessel having a jacket temperature of the reactor set at 25 °C under nitrogen, and the internal temperature was raised to 38 °C. 3.1 M HCl (6.4 kg) in CPME was added, and as a result, based on the assay of Compound 1 in the crude Compound 2 and the assay of HCl in the crude Compound 2, a total of 1.07 equivalents of HCl was achieved. Next, n-heptane (139.4 kg) was added and the internal reaction temperature was raised to 51 °C. Next, at 50 °C, a seed crystal slurry of Compound 2 in CPME / n-heptane (291 g, 0.87 wt%) was added and this temperature was maintained for 105 minutes. Next, the opaque reaction slurry was cooled to 11 °C over 12 hours at a linear cooling rate. Next, using a filter dryer, the slurry was filtered under vacuum at 9 °C. Next, 20 volume% CPME in n-heptane (57.4 kg of CPME, 180 kg of n-heptane) was added to the reaction vessel and cooled to 11 °C. Next, half of the mixture was poured into the filter dryer as a chromatography wash solution and flowed through. The latter half was passed through the filter dryer as a slurry wash solution. Next, when the wet filter cake was dried under vacuum at stepwise jacket temperatures of 25, 35, 46, 54 °C, Compound 2 was obtained in a 64% yield (from Compound 1F) with a purity of 99.6 area% and residual solvents of 0.3% w of CPME and <0.1% w of n-heptane. (Example 10) Further Preparation of Compound 2

[0421] Compound 1 in solution was further diluted with n-heptane and heated to 40 °C. At this temperature, approximately 3 M HCl in CPME (1.1 meq with respect to Compound 1F) was added. This solution was further heated to 48 - 53 °C and a second portion of n-heptane was charged. For optional seed crystal addition with seed crystals of Compound 2, this clear solution was cooled to 53 °C (seed crystal addition is optional but preferred in the manufacturing situation). When seed crystal addition was performed, the solution was salt decomposed at 53 °C and then cooled to 10 °C over 12 hours.

[0422] Performing a crystallization aging procedure (a procedure of repeating heating and cooling cycles) can improve the color of the obtained compound 2 after filtration. The suspension of the product was filtered, washed once with cold CPME / n-heptane (20:80 volume) and once with cold n-heptane, and then dried in vacuo. (Example 11) (2R,4S)-4-{[3,5-Bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (Compound 3)

Chemical Structure

[0423] To isopropyl acetate (IPAC, 214 kg, 6.11 wt%) in an inert reactor was added ethyl (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate hydrochloride (Compound 2, 35.0 kg, 48.4 mol), and the mixture was stirred at 22 °C until dissolution was achieved. Deionized water (245 kg, 7.00 wt%) was added, and the reaction mixture was stirred at 23 °C for 35 minutes. Then, stirring was stopped and the phases were separated, and the lower aqueous phase was removed. The process of adding deionized water (245 kg, 7 wt%), stirring, and removing the lower aqueous phase was repeated three more times. Next, the organic phase was concentrated under reduced pressure to approximately 71 L (approximately 2 volumes) while maintaining an internal temperature of 55 °C or less. Next, ethanol (115 kg, 3.29 wt%) was added, and the reaction mixture was concentrated under reduced pressure to approximately 78 L (approximately 2 volumes) while maintaining an internal temperature of 55 °C or less. The process of adding ethanol (115 kg, 3.29 wt%) and concentrating was repeated two more times. Next, the reaction mixture was cooled to 25 °C and treated with charcoal by cartridge. Next, the cartridge was rinsed with ethanol (100 kg, 2.86 wt%) and concentrated in vacuo at 55 °C or less to 147 L (approximately 3.8 volumes), and then, when 35 L of EtOH (1.0 volume) was added, the free base form of ethyl (2R,4S)-4-{[3,5-bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylate (Compound 1) in ethanol was obtained. Next, to the reaction vessel containing Compound 1 in ethanol, a 14 wt% NaOH solution (15.8 kg, 1.13 eq) was added while maintaining a reaction temperature of 20 °C. The reaction mixture was stirred at 20 °C for 5 hours to achieve complete conversion.

[0424] 34 wt% calcium chloride (aqueous) (10.8 kg) was added to the inert reactor. Next, deionized water (336 L, 9.61 wt. with respect to Compound 1) and ethyl acetate (15 kg, 0.43 wt. with respect to Compound 1) were added and the mixture was stirred for 30 minutes to obtain "Solution B".

[0425] Next, while stirring Solution B, it was cooled to 9 °C. Next, while maintaining the temperature of Solution B at 10 °C, Solution A (see above) was added via a filter over 90 minutes. Next, the container of Solution A was rinsed into Solution B with additional ethanol (50 kg, 1.43 wt. with respect to Compound 1). The resulting slurry was stirred at 9 °C for 1 hour. Next, the solid was collected by filtration and rinsed with deionized water (2 x 175 kg, 5 wt. with respect to Compound 1). Next, the solid was dried in vacuo at 50 °C for 21 hours to obtain 27.6 kg of amorphous obeticholic acid hemicalcium (Compound 3) containing < 1 wt% water (77% yield, based on the number of moles of Compound 2). Compound 3 was reworked as described in Example 12 below. (Example 12) Rework of Compound 3

[0426] Compound 3 (27.6 kg) was dissolved in ethanol (55.2 kg, 2 wt% relative to Compound 3) at 45 - 48 °C, and then cooled to 11 °C. The solution was filtered and added to a pre-cooled (approximately 10 °C) mixture of an aqueous CaCl2 solution (33 - 35 wt% of 8.2 kg, 0.3 wt%), water (262 kg, 9.5 wt%) and ethyl acetate (12.6 kg, 0.46 wt%). The resulting suspension was filtered off, washed with water (2×5 wt%, 138 kg per washing step), and the solid was dried in vacuo while maintaining an internal temperature of 45 °C or less for 23 hours, to obtain 24.8 kg (91% yield) of an amorphous hemicalcium salt of (2R,4S)-4-{[(3,5-bis(trifluoromethyl)benzyl]-[5-(3-carboxypropoxy)pyrimidin-2-yl]amino}-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (Compound 3) containing <1 wt% water, with a purity of 97.5 wt% and >99.9 area%. (Example 13) Further preparation of Compound 3

[0427] Compound 2 was neutralized and dissolved in aqueous NaOH in EtOH. This solution was filtered through activated carbon. Vacuum distillation was carried out to concentrate the solution. For the saponification of the ester, an aqueous NaOH solution was added to obtain the sodium salt of Compound 1 in the solution, and the ester was formed in this step and the previous step.

[0428] Subsequently, a mixture of an aqueous CaCl2 solution and EtOAc was prepared in a second container. Next, the Na salt of Compound 1 from the first container was added to this mixture, whereby Compound 3 precipitated. If necessary, the suspension may be heated to NMT at 25 °C and then cooled to 8 °C. The solid Compound 3 was filtered off at 8 °C, washed with water, and dried in vacuo. (Example 14) Milling of recycled Compound 3

[0429] Compound 3 was jet milled using an 8-inch spiral mill. The feed rate, venturi pressure, and mill pressure were adjusted within the ranges listed below to produce micron-sized Compound 3 in accordance with the particle size acceptance criteria (D90 = 6 - 15 μm). Feed rate: 17 - 20 kg / hour Mill pressure: 20 PSI / 1.4 bar Venturi pressure: 100 PSI / 6.9 bar Processing gas: Nitrogen Analysis: Mastersizer 3000. (Example 15) Jet milling of another preparation of Compound 3 The particle size distribution was adjusted to the target parameters d90: 6 - 15 microns by micronizing with a spiral jet mill, 8-inch jet mill, 8005, and KT4 LIW feeder. Three samples were jet milled and the following results were obtained: d90: 8 microns, 8 microns, and 9 microns d50: 4 microns, 3 microns, and 4 microns d10: 2 microns, 1 micron, 1 micron (Example 16) Crystalline obcetrapib hemicalcium

[0430] 2 g of amorphous obcetrapib hemicalcium was added to acetonitrile (ACN) / methyl tert-butyl ketone (MIBK) (6:1 ratio at 200 mg / ml) and the sample was heated to 50 °C for 5 minutes until all of the solid had dissolved. The sample was then placed in a water bath and cooled from 50 °C to 5 °C at 0.9 °C / min over 48 hours. The sample was maintained at 5 °C for 3 days and then transferred to -20 °C for 30 minutes before isolating the solid. The solid was air dried for 2 hours before further characterization. This process resulted in the formation of crystalline obcetrapib hemicalcium. (Example 17) Polarized light microscopy (PLM)

[0431] Polarized microscope photographs were captured at room temperature using a Nikon DS-Fi2 upright microscope. The sample (2 mg) was mounted on a slide glass and covered with a drop of silicone oil using a cover glass on top of the sample for analysis. The sample was not protected from light. (Example 18) X-ray powder diffraction (XRPD)

[0432] In a silicon zero-background holder, a Panalytical X’Pert was used with an incident beam of Cu radiation generated using an Empyran tube and a fine focus source. 3 XRPD was performed using a powder diffractometer. Before analysis, a silicon standard (NIST SRM 640d) was analyzed to confirm that the Si 111 peak position was consistent with the certified position by NIST. Approximately 5 - 10 mg of the sample was placed in a silicon zero-background holder and manually flattened using an aluminum spatula to minimize the height difference across the sample. Next, this holder was mounted on the analytical instrument. The XRPD parameters used are listed in Table 10.

Table 10

[0433] An X-ray powder diffractometer manufactured by PANalytical was used under the following measurement conditions. Data was acquired using DataViewer and evaluated using X’Pert High Score Plus: X-ray tube Cu LFF HR Configuration Transmission type X-ray mirror Focusing X-ray mirror W / Si Solar slit 0.02 rad Detector Pixel 1D Detector effective length 1.69° Divergence slit Fixed Divergence slit size 1 / 2° X-ray tube excitation 40 mA, 40 kV 2Theta range 2° to 40° Measurement mode: Continuous Time per step: 300 seconds Step size: 0.013° (2Theta) Rotation speed: 1 rotation per second (Example 20) X-ray powder diffraction pattern

[0434] The diffraction pattern related to Figure 3 was measured using an Empyrean powder diffractometer manufactured by Malvern PANalytical in transmission mode. The sample was prepared as a thin layer between two Kapton foils and measured in continuous mode. The detector measures from approximately 2° (2θ) to 40° (2θ). Signal peaks can be observed at approximately 3.4° (2θ), approximately 7.0° (2θ), and approximately 9.2° (2θ). The peak at approximately 5.6° (2θ) is attributed to the Kapton foil. (Example 21) X-ray powder diffraction method for crystalline obeticholic acid HCl / Compound 1D

[0435] The diffraction pattern was measured using a Thermo Fisher Scientific ARL Equinox 1000 powder diffractometer. The diffractometer is equipped with a copper source, a germanium (111) monochromator for providing monochromatic Cu Kα1 radiation, and a position-sensitive gas ionization detector.

[0436] The sample was measured in reflection mode using an Al sample holder without any further preparation (i.e., grinding). The detector measures simultaneously over the full angular range from approximately 2° (2θ) to 120° (2θ). For HCl obeticholic acid, distinguishable signals useful for phase identification are observed up to approximately 45° (2θ). The temperature of the diffractometer is usually set to approximately 30°C during the measurement. (Example 22) X-ray powder diffraction method for crystalline obeticholic acid HCl

[0437] A Rigaku SmartLab X-ray diffractometer was configured in a Bragg-Brentano reflection configuration equipped with a beam stop and a knife edge to reduce the incident beam and air scattering. The data collection parameters are shown in Table 11.

Table 11

[0438] The FTIR spectrum of a sample of amorphous obeticholic acid hemicalcium is described in Figure 4. The FTIR spectrum was obtained using a Bruker Tensor 27 spectrometer equipped with a Platinum ATR-QL-Diamond unit. The milled sample was placed in the ATR unit without any pretreatment. (Example 24) 1 H-NMR spectroscopy

[0439] The NMR spectrum of a solution prepared from a sample of amorphous obeticholic acid hemicalcium is described in Figure 5. The NMR spectrum was obtained using a 600 MHz AVANCE NEO Bruker, using tetramethylsilane (TMS) as an internal standard with respect to the chemical shift at 0.0 ppm, in deuterated MeOH as the solvent. The shift of the spectrum is consistent with the chemical structure. (Example 25) Modulated differential scanning calorimetry (mDSC)

[0440] A sample containing an mDSC thermogram is described in Figure 12, and Figure 14 was prepared using a T-zero aluminum pan with a pinhole. The gradient rate was modulated at ±0.5 °C every 60 seconds, from 25 °C to 225 °C at 2 °C / min. The instrument used was a TA Q2500 DSC manufactured by TA Instruments. (Example 26) Modulated differential scanning calorimetry (mDSC)

[0441] The DSC2500 manufactured by TA Instruments was used. The starting temperature was 25 °C, and the sample was heated at 2 °C per minute and modulated by ±0.5 °C every 60 seconds up to 225 °C. A T-zero aluminum pan and a T-zero airtight lid with additional factory pinholes punched and expanded were used for this test. The integrated thermogram (displaying reversible heat flow) derived from the sample is included in Figure 13. The sample showed a glass transition with a Tg of approximately 111 °C. (Example 27) Method for evaluating stability

[0442] The stability study of obeticholic acid in its crystalline and amorphous forms was conducted at 70 °C / 75% relative humidity (RH). This solid was placed in a 4.0 mL glass vial without a stopper (open condition) and stored at 70 °C / 75% RH. At the time points of day 1 (24 hours) and day 7, the samples were removed from the stability chamber. The physical stability of this solid was analyzed by XRPD, and the chemical purity was analyzed by HPLC. The samples collected at each time point were dissolved in methanol and then analyzed by HPLC. To minimize the effect of adsorption of potential analytes on the filter, the first 0.5 mL of supernatant passing through the filter was discarded, and then the samples for HPLC analysis were collected. The purity for each sample was determined based on the peak area percentage and compared with the sample at T = 0. (Example 28) Method used to evaluate the dynamic solubility in a biorelevant medium

[0443] Dynamic solubility studies were conducted on the crystalline and amorphous forms of obicetrapib in biorelevant media including simulated intestinal fluid at pH 5.0 (FeSSIF) and fasted simulated intestinal fluid at pH 6.5 (FaSSIF) at 37°C. Solids were magnetically stirred at 600 RPM in a shaker bath, and samples were withdrawn with a 1.0 mL syringe at T = 15 minutes, 30 minutes, 60 minutes, 90 minutes, and 120 minutes. Solubility was measured using the HPLC method presented by the requester. The compound was added to a 4.0 mL glass vial at approximately 20.0 mg / mL. The sample was stirred with a vortex mixer for about 5 minutes to confirm the presence of undissolved excess powder. Samples taken at each time point were centrifuged at 1200 RPM, filtered using a 0.45 μm polytetrafluoroethylene (PTFE) filter, diluted with methanol, and then analyzed by HPLC. To minimize the effect of potential analyte adsorption on the filter, the first 0.5 mL of supernatant passing through the filter was discarded, and then samples for HPLC analysis were taken.

[0444] The present invention has been particularly shown and described with reference to preferred embodiments and various alternative embodiments, but it will be understood by those skilled in the relevant art that various changes in form and detail may be made in this embodiment without departing from the spirit and scope of the present invention. (Example 29) Preparation of Form A of Crystalline Obicetrapib Hydrochloride

[0445] Obicetrapib HCl (approximately 43 mg, prepared in accordance with the disclosure herein) was combined with CPME / heptane (1:7) (0.8 mL) in a 1 dram vial, and this mixture was magnetically stirred at room temperature. After 2 weeks, the solid was separated by centrifugation, and the residual liquid was removed with a pipette. The sample was dried in a vacuum desiccator for approximately 30 minutes to obtain approximately 25 mg of Form A. (Example 30) Preparation of Form B of Crystalline Obicetrapib Hydrochloride

[0446] Obcetrapib HCl (approximately 84 mg, prepared according to the disclosure herein) was dissolved in 3.5 mL of toluene. The solution was stirred at room temperature and approximately 1 molar equivalent of hydrochloric acid (116 μL, 1 M solution in diethyl ether) was added to the above solution. Next, heptane (4 mL) was added to this mixture and a cloudy solution was obtained within a few minutes. After stirring overnight, the suspension was filtered under vacuum and the solid was dried on the filter under reduced pressure for a short time to obtain approximately 40 mg of Form B (weighed 26 days after ambient storage). (Example 31) Preparation of crystalline obcetrapib hydrochloride Form C

[0447] Obcetrapib HCl (approximately 84 mg, prepared according to the disclosure herein) was dissolved in 1.0 mL of isopropyl acetate. The solution was stirred at room temperature and approximately 1 molar equivalent of hydrochloric acid (116 μL, 1 M solution in diethyl ether) was added to the above solution. Next, heptane (6 mL) was added to this mixture and a cloudy solution was obtained. After stirring overnight, the suspension was filtered under vacuum and the solid was dried on the filter under reduced pressure for a short time to obtain approximately 25 mg of Form C (weighed 26 days after ambient storage). (Example 32) Preparation of crystalline obcetrapib hydrochloride Form D

[0448] Obcetrapib HCl (approximately 46 mg, prepared according to the disclosure herein) was combined with butyl acetate / heptane (1:5) (0.6 mL) in a dram vial and the mixture was magnetically stirred at room temperature. After 2 weeks, the solid was separated by centrifugation and the residual liquid was removed by pipette. The sample was dried in a vacuum desiccator for approximately 30 minutes to obtain approximately 15 mg of Form D. (Example 33) Preparation of single crystal sample

[0449] Heat to approximately 60 °C and dissolve obicetrapib HCl (approximately 10 mg, prepared according to the disclosure herein) in a mixture of cyclopentyl methyl ether and heptane (1:8) (0.232 mL). Cool the resulting solution to 50 - 55 °C and let stand at this temperature overnight. The next day, some needle-shaped solids were observed on the side of the vial. A temperature cycling experiment by continuous heating - cooling was performed: The sample was heated to 55 - 60 °C and then cooled to 45 - 50 °C and held at that temperature for several hours. Approximately 5 days later, aggregated long blade-like objects were observed under the microscope and were found to be of sufficient size and quality. (Example 34) Solution of single crystal structure

[0450] Colorless long blade-shaped crystals of the single crystal of Example 33 having a formula [4(C 32 H 32 F9N4O5)·2(C 32 H 31 F9N4O5)·C6H 12 O·C7H 16 ·4(Cl)·[+ solvent]] with approximate dimensions of 0.02 × 0.09 × 0.28 mm were mounted in a random orientation on a Mitegen micromesh mount. Data were collected from a shock-cooled single crystal at 150(2) K using a Bruker AXS D8 Quest four-circle diffractometer equipped with an I-mu-S microsource X-ray tube, a lateral step multilayer (Goebel) mirror as a monochromator, and a PhotonIII_C14 charge-integrating photon counting pixel array detector. The diffractometer used CuK αRadiation (λ = 1.54178 Å) was used. All data were integrated using SAINT V8.40B, and multi-scan absorption correction was applied using SADABS 2016 / 2 (Bruker, SAINT, V8.40B, Bruker AXS Inc., Madison, Wisconsin, USA; L. Krause, R. Herbst-Irmer, G. M. Sheldrick, D. Stalke, J. Appl. Cryst. 2015, 48, 3-10, doi:10.1107 / S1600576714022985). The structure was solved by the dual method with SHELXT and refined by full-matrix least-squares method against F 2 for (G.M. Sheldrick, Acta Cryst. 2015, A71, 3-8, doi:10.1107 / S2053273314026370; G. M. Sheldrick, Acta Cryst. 2015, C71, 3-8, doi:10.1107 / S2053229614024218). All non-hydrogen atoms were refined with anisotropic displacement parameters. Hydrogen atoms bonded to carbon, hydroxyl H atoms of carboxylic acids, and H atoms of the most planar (sp2 hybridized) N-H groups were refined isotropically against calculated positions using a riding model. For further details of hydrogen atom treatment, methyl CH3 and hydroxyl H atoms were rotated but not tilted to best fit the experimental electron density. The Uiso values were constrained to 1.5 times Ueq of their pivot atoms for methyl and hydroxyl groups and 1.2 times for all other hydrogen atoms.

[0451] The asymmetric part of the structure consists of six major organic molecules, four chloride anions and some solvation molecules (methylcyclopentyl ether and heptane). Four of the six major fragments are cationic and two are neutral molecules ("free bases"). Protonation occurs at the "N3" nitrogen atom of the pyrimidine ring for all four cations, all four N-H+ units are ionized, and all four chloride anions are ionized. That is, two are associated with two N-H+ units and the others are associated with the carboxyl moiety. The equivalent positions for the two free base molecules are not protonated and no close contacts to potential H-bond acceptors are observed.

[0452] Widespread irregularities are observed throughout the structure and some fragments show very large thermal vibrations. This is even more pronounced in the case of the two free base molecules (residues 5 and 6), especially their bis(trifluoromethyl)benzene moieties. The solvation molecules are irregular over a wide range and also show very large thermal vibrations and are only partially resolved.

[0453] All six major obeticholic acid molecules (cation and free base) were constrained to have similar geometric shapes. Minor irregularities were constrained to have similar geometric shapes as well-defined fragments of another molecule's non-irregularities (using the SAME command in Shelxl). All C-F bond distances and all F-C-F angles were each constrained to be similar to one another. The carbon atoms of the major and minor portions of the bis(trifluoromethyl)benzene moiety of residues 1, 2, and 3, and the mon(trifluoromethyl)benzene moiety of residue 3 were constrained to be near planar (using the FLAT command in Shelxl). The latter atomic displacement parameters ("ADP") were constrained to approach isotropy. The Uij components of the ADP for atoms within 2.0 Å of one another were constrained to be similar. The position of one H atom of an N-H+ (H3A_3) was refined. The hydroxyl H atoms of the carboxylic acid groups were rotated. Some were further constrained based on hydrogen bonding considerations. A mild damping factor was applied during the first refinement cycle. In the final refinement cycle, some hydroxyl H atoms were set to ride on the O atoms to which they are attached (H4_1, H4_4, H4B_1), excluding the damping factor.

[0454] Irregularities were refined against the following fragments and occupancies were refined as follows according to the above conditions: · The bis(trifluoromethyl)benzene moiety of residue 1 (cation). Occupancy: 0.589(14) - 0.411(14). · The bis(trifluoromethyl)benzene moiety of residue 2 (cation). Occupancy: 0.534(11) - 0.466(11). · The bis(trifluoromethyl)benzene and mon(trifluoromethyl)benzene moieties of residue 2 (cation). Occupancies: 0.598(11) - 0.402(11) and 0.492(19) - 0.492(19). · The 4-(pyrimidin-2-yloxy)butanoic acid fragment of residue 6 (free base). Occupancy: 0.493(11) - 0.507(11).

[0455] The methylcyclopentyl ether solvated molecules were refined to full occupancy. Bond distances and angles were restrained to their expected target values, and the Uij components of the ADP for atoms closer than 2.0 Å to each other were similarly restrained. The ADP was restrained to approach isotropy. The heptane solvated molecules were refined to be disordered in two directions. Bond distances and angles were restrained to their expected target values, and the Uij components of the ADP for atoms closer than 2.0 Å to each other were similarly restrained. A mild anti-attenuation restraint was applied to avoid close contacts with the atoms of the main molecule. Following these conditions, the occupancy was refined to 0.460(13)–0.540(13).

[0456] The structure contains a further solvent-accessible void volume of 980 Å 3 . The two major void spaces (333 Å 3 each) are likely to contain poorly defined and very disordered solvated molecules. No large electron density peaks were observed in the solvent-accessible voids (less than 0.70 electrons per Å 3 ), and the remaining electron density peaks are not arranged in a recognizable pattern. Instead, the SQUEEZE routine implemented in the program Platon (A. L. Spek J. Appl. Cryst. 2003, 36, 7–13) was used to expand the structure factors by the inverse Fourier transform method (P. van der Sluis, & A. L. Spek, Acta Cryst. 1990, A46, 194–201). This SQUEEZE procedure accounts for 229 electrons in this volume, i.e., approximately one heptane molecule (100.2 electrons / heptane) per each of the two major void spaces.

[0457] The Flack x parameter was determined using Parsons' method with 4528 quotients [(I+)-(I-)] / [(I+)+(I-)], and refined to 0.046(17) (S. Parsons, H. Flack, T. Wagner, Acta Cryst. 2013, B69, 249–259). (Example 35) X-ray powder diffraction pattern

[0458] A Rigaku SmartLab X-ray diffractometer was configured in a Bragg-Brentano reflection configuration equipped with a beam stop and a knife edge to reduce the incident beam and air scattering. The data collection parameters are shown in the following table (Table 12). This method has not been verified.

Table 12

[0459] The X-ray powder diffraction pattern was calculated from the single crystal structure solution of Example 34. The pattern was generated using commercially available software called Mercury 3.3 (Build RC5) from the Cambridge Crystallographic Data Centre (CCDC).

[0460] A table of peaks related to the calculated pattern is presented in Table 13 below.

Table 13-1

Table 13-2

Table 13-3

[0461] All references, issued patents, and patent applications cited within the body of this specification are hereby incorporated by reference in their entirety for all purposes.

Claims

1. Crystalline ovicetrapib hydrochloride.

2. The crystalline ovicetrapib hydrochloride according to claim 1, which is in solvate form.

3. The solvate of crystalline ovicetrapib hydrochloride according to claim 2, wherein the solvate comprises an organic solvent.

4. The solvate of crystalline ovicetrapib hydrochloride according to claim 3, wherein the organic solvent comprises an organic solvent selected from methanol, ethanol, isopropanol, acetic acid, acetonitrile, acetone, methyl isobutyl ketone, isopropyl acetate, tetrahydrofuran, methyl t-butyl ether, cyclopentyl methyl ether, N-methyl-2-pyrrolidone, dimethyl sulfoxide, dimethylformamide, 2-methyl-tetrahydrofuran, dichloromethane, 1,4-dioxane, 1,2-difluorobenzene, toluene, heptane, and hexafluoroisopropanol.

5. The solvate of crystalline ovicetrapib hydrochloride according to claim 4, wherein the organic solvent comprises an organic solvent selected from cyclopentyl methyl ether, toluene, and heptane.

6. The solvate of crystalline ovicetrapib hydrochloride according to claim 2, wherein the solvent comprises cyclopentyl methyl ether and heptane.

7. The solvate of crystalline ovicetrapib hydrochloride according to claim 3, further comprising one or more of ovicetrapib free base and hydrochloric acid.

8. The solvate of crystalline ovicetrapib hydrochloride according to claim 6, further comprising one or more of ovicetrapib free base and hydrochloric acid.

9. The crystalline ovicetrapib hydrochloride according to claim 1, having an X-ray powder diffraction pattern including a peak at approximately 9.8° 2θ.

10. The crystalline ovicetrapib hydrochloride according to claim 1, having an X-ray powder diffraction pattern comprising one or more peaks selected from approximately 8.1°²θ, approximately 9.8°²θ, approximately 13.8°²θ, approximately 16.7°²θ, and approximately 19.5°²θ.

11. The crystalline ovicetrapib hydrochloride according to claim 1, having substantially the same X-ray powder diffraction pattern as the X-ray powder diffraction pattern of Figure 19.

12. Crystalline ovicetrapib hydrochloride of form A.

13. Crystalline ovicetrapib hydrochloride of form A according to claim 12, having an X-ray powder diffraction pattern including a peak at approximately 8.6°²θ, two peaks between approximately 9.7°²θ and approximately 10.4°²θ, and a peak at approximately 9.0°²θ.

14. Crystalline ovicetrapib hydrochloride of form A according to claim 12, having substantially the same X-ray powder diffraction pattern as the X-ray powder diffraction pattern of Figure 22.

15. Crystalline ovicetrapib hydrochloride of form B.

16. Crystalline ovicetrapib hydrochloride of form B according to claim 15, having an X-ray powder diffraction pattern including peaks at approximately 6.5°2θ, approximately 8.8°2θ, and approximately 11.0°2θ.

17. Crystalline ovicetrapib hydrochloride of form B according to claim 15, having substantially the same X-ray powder diffraction pattern as the X-ray powder diffraction pattern of Figure 23.

18. Crystalline ovicetrapib hydrochloride of form C.

19. Crystalline ovicetrapib hydrochloride of form C according to claim 18, having substantially the same X-ray powder diffraction pattern as the X-ray powder diffraction pattern of Figure 24.

20. Crystalline ovicetrapib hydrochloride of form D.

21. Crystalline ovicetrapib hydrochloride of form D according to claim 20, having substantially the same X-ray powder diffraction pattern as the X-ray powder diffraction pattern of Figure 25.

22. The crystalline ovicetrapib hydrochloride according to claim 1, wherein the weight percentage of hydrochloric acid is between about 0.01% and about 8%.

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