ES2859664T3 - Solid state forms of sofosbuvir - Google Patents

Abstract

Crystalline form of Sofosbuvir designated Form E and characterized by data selected from one or more of the following: an X-ray powder diffraction pattern having peaks at: 12.4, 16.2, 17.2, 25.0, and 25.3 degrees two theta ± 0.1 degrees two theta; an X-ray powder diffraction pattern as illustrated in Figure 1 or Figure 7; an X-ray powder diffraction pattern having peaks at: 12.4, 16.2, 17.2, 25.0 and 25.3 degrees two theta ± 0.1 degrees two theta and no peaks at: 10 , 9 and 14.2 degrees two theta ± 0.2 degrees two theta, and combinations of these data.

Description

DESCRIPTION

Solid state forms of sofosbuvir

Field of the invention

The present invention encompasses a solid state form of Sofosbuvir and pharmaceutical compositions thereof. Background of the invention

Sofosbuvir, L-alanine, N - [[P (S), 2'R] -2'-deoxy-2'-fluoro-2'-methyl-P-phenyl-5'-uridylyl] -, 1-methylethyl ester , or (2S) -isopropyl 2 - (((((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) -4-fluoro- 3-hydroxy-4-methyltetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) amino) propanoate, which has the following formula,

is a second-generation, orally available nucleoside analog that inhibits the NS-5 protein of the hepatitis C virus (HCV). Sofosbuvir and its isomer act as prodrugs and are converted through a series of in vivo transformations to an active triphosphate metabolite.

Sofosbuvir is marketed under the registered trade name SOVALDI ®. SOVALDI ® is available as immediate release tablets. Each tablet contains 400 mg of sofosbuvir. The active ingredient loading in SOVALDI ® tablets is approximately 30% and the tablets also contain glidant. More specifically, SOVALDI ® tablets include the following active ingredients: colloidal silicon dioxide, croscarmellose sodium, magnesium stearate, mannitol and microcrystalline cellulose. The tablets are film-coated with a coating material containing the following active substances: polyethylene glycol, polyvinyl alcohol, talc, titanium dioxide and yellow iron oxide. WO 2013/082003 and WO 2014/120981 also describe tablets containing sofosbuvir.

Sofosbuvir is described in US 7,964,580 and US 8,334,270. Solid state forms of Sofosbuvir are described in WO 2010/135569, US 2011/251152, WO 2011/123645 and CN 104130302. CN 104277088 and CN 104447924 also describe crystalline forms of sofosbuvir. Polymorphism, the appearance of different crystalline forms, is a property of some molecules and molecular complexes. A single molecule can cause a variety of polymorphisms that have different crystal structures and physical properties such as melting point, thermal behaviors (for example, measured by thermogravimetric analysis - "TGA", or differential scanning calorimetry - "DSC"), pattern X-ray diffraction, infrared absorption fingerprint and solid state (13C-) NMR spectrum. One or more of these techniques can be used to distinguish different polymorphic forms of a compound.

US 2011/251152 describes a number of crystalline forms of Sofosbuvir, that is, forms 1, 2, 3, 4, 5 and 6, which are characterized by X-ray powder diffraction peaks (DPRX), as well as a amorphous form. According to this publication, the crystalline forms 2, 3, 4 and 5 of Sofosbuvir are said to be prepared by crystallization of dichloromethane, chloroform, acetonitrile and anisole. However, after filtration and / or drying, these crystalline forms form conversion to Form 1.

CN 104130302 describes Form A crystalline sofosbuvir characterized by Form A XRD is described as a non-hydrated, non-solvated form. Form A is prepared by dissolving Sofosbuvir in a solvent and adding an antisolvent and allowing the mixture to sit in a sealed container for 15-24 hours under certain conditions. Solvent / antisolvent combinations include anhydrous ethanol and one of isopropyl ether, cyclohexane, n-pentane, or toluene, or the solvent / antisolvent can be acetone / n-pentane, acetone / petroleum ether, or ethyl acetate / ether. Petroleum.

Different salts and solid state forms (including solvated forms) of a pharmaceutical active ingredient can have different properties. Such variations in the properties of various salts and solid state forms and solvates can provide a basis for improving the formulation, for example, facilitating better processing or manipulation of the characteristics, changing the dissolution profile in a favorable direction, or improving stability. (polymorphic as well as chemical stability) and shelf life. These variations in the properties of different salts and solid state forms also offer improvements to the final dosage form, for example if they serve to improve bioavailability. Different salts and solid state forms and solvates of a pharmaceutical active ingredient as well they can elicit a variety of polymorphs and crystalline forms, which may, in turn, provide additional opportunities to evaluate variations in the properties and characteristics of a solid pharmaceutical active ingredient. The discovery of new solid state forms and solvates of a pharmaceutical product can provide materials that have desirable processing properties, such as easy to handle, easy to process, storage stability and ease of purification or as desirable intermediate crystal forms that facilitate conversion to other polymorphic forms. New solid state forms of a pharmaceutically useful compound may also provide an opportunity to improve the performance characteristics of a pharmaceutical product. Enlarges the repertoire of materials that a formulation scientist has available for formulation optimization, for example, they provide a product with different properties, for example, a different crystalline habit, superior crystallinity or polymorphic stability that can provide better processing or characteristics of handling, improved dissolution profile or improved shelf life (chemical / physical stability). For example, Sofosbuvir Form 6 has now been found to have a high tendency to become electrostatically charged. Electrostatically charged active pharmaceutical ingredients may show a poor fluidity and / or tendency to stick and thus may ultimately result in complications during the manufacturing process operations of a pharmaceutical composition based on such electrostatically charged pharmaceutical active ingredient. . Furthermore, poor content uniformity can be observed in the final dosage form when using a dry process such as, for example, dry compression to make a pharmaceutical composition with an electrostatically charged pharmaceutical active ingredient. For at least these reasons, there is a need for additional solid state forms (including solvated forms) of Sofosbuvir. Additionally, prior art processes for producing crystalline forms of Sofosbuvir, such as those described above, are generally impractical for the large scale preparation environment. On the other hand, the resulting products can undergo polymorphic transformations, which can lead to polymorphically impure materials. Therefore, there is a need in the art to provide additional processes for preparing solid state forms of Sofosbuvir.

The present invention aims to provide a new crystalline form of Sofosbuvir.

Summary of the invention

The present invention provides a solid state form of Sofosbuvir and pharmaceutical compositions thereof. In particular, the present invention provides a crystalline form of Sofosbuvir designated Form E and characterized by data selected from one or more of the following: an X-ray powder diffraction pattern having peaks at: 12.4, 16.2, 17.2, 25.0 and 25.3 degrees two theta ± 0.1 degrees two theta; an X-ray powder diffraction pattern as illustrated in Figure 1 or Figure 7; an X-ray powder diffraction pattern having peaks at: 12.4, 16.2, 17.2, 25.0 and 25.3 degrees two theta ± 0.1 degrees two theta and no peaks at: 10 , 9 and 14.2 degrees two theta ± 0.2 degrees two theta, and combinations of these data.

The present invention also encompasses the use of Sofosbuvir Form E for the preparation of Sofosbuvir pharmaceutical compositions.

The present invention comprises processes for preparing the aforementioned pharmaceutical compositions. The process comprises combining Sofosbuvir Form E with at least one pharmaceutically acceptable excipient.

Solid state Form E of Sofosbuvir and pharmaceutical compositions thereof can be used as medicaments, in particular, for the treatment of Hepatitis C.

Brief description of the drawings

Figure 1 shows a Form E X-ray powder diffractogram of Sofosbuvir.

Figure 2 shows a Form 1 X-ray powder diffractogram of Sofosbuvir.

Figure 3 shows a Form 6 X-ray powder diffractogram of Sofosbuvir

Figure 4 shows the DVS Isothermal Plot of Sofosbuvir Form 1

Figure 5 shows the DVS Isothermal Plot of Sofosbuvir Form 6

Figure 6 shows the DVS Isothermal Plot of Sofosbuvir Form E, prepared according to Example 7

Example 7 - DPRX of Sofosbuvir Form E in 2-30 degree 2-theta interval

Figure 8 shows a Form 7 X-ray powder diffractogram of Sofosbuvir

Figure 9 provides the peaks of the Sofosbuvir Form 7 X-ray powder diffractogram.

Figure 10 shows an X-ray powder diffractogram of Sofosbuvir Form 7 (Example 1) measured using Ka1 / Ka2 copper radiation with a wavelength weighted median of 1.54187 A.

Figure 11 shows the X-ray powder diffraction peaks for sofosbuvir Form 7 (Example 1) and the relative intensities measured using copper Ka1 / Ka2 radiation with a wavelength weighted median of 1.54187 A. In In this table, 2-theta values are reported to within 0.2 degrees 2-theta.

Figure 12 shows the sofosbuvir Form 1 X-ray powder diffraction peaks and relative intensities. In this table, 2-theta values are reported to within 0.2 degrees 2-theta.

Figure 13 shows the dissolution curves of Form 7 versus Form 6 measured in 50 mM phosphate buffer (pH 6.8, 900 ml) at 37 ° C, using USP apparatus II, paddle method according to the example described below.

Detailed description of the invention

The present invention encompasses a solid state form of Sofosbuvir. The solid state properties of Sofosbuvir can be influenced by controlling the conditions under which Sofosbuvir is obtained in solid form.

In some embodiments, the crystalline / solid state form of the Sofosbuvir of the invention is substantially free from any other form of Sfosbuvir or from specified polymorphic forms of Sofosbuvir, respectively.

As used herein, "substantially free" refers to that the solid state forms of the present invention contain 20% (w / w) or less, 10% or less, 5% or less, 2% or less or 1% or less of any other form of the object compound or of a specified polymorph of Sfosbuvir as measured, for example, by DRXP, According to some embodiments, the salts and solid state forms of the present invention contain the 10% (w / w) or less, 5% (w / w) or less, 2% (w / w) or less, 1% (w / w) or less, 0.5% (w / w) or less or 0.2% (w / w) or less of polymorphs or a specified polymorph of Sofosbuvir. In other embodiments, the solid state form of Sofosbuvir of the present invention contains 1% to 20% (w / w), 5% to 20% (w / w), or 5% to 10% (w / w ) in any solid state form of a specified polymorph of Sofosbuvir.

Depending on what comparison to other solid state forms is made, the solid / crystalline form of Sofosbuvir of the present invention has advantageous properties selected from at least one of the following: chemical purity, fluidity, solubility, dissolution rate, morphology and crystalline habit, stability - such as chemical as well as thermal and mechanical stability with respect to polymorphic conversion, stability towards hydration and / or storage stability, low residual solvent content, a lower degree of hygroscopicity, flowability and advantageous processing and characteristics handling such as compressibility and bulk density.

A solid state form, such as a crystal form or amorphous form, may be referred to herein as being characterized by graphical data "as illustrated in" or "as substantially illustrated in" a Figure. Such data includes, for example, X-ray powder diffractograms. As is well known in the art, graphical data potentially provides additional technical information to further define the respective solid state form (a so-called "fingerprint") which may not necessarily be described by reference to numerical values or peak positions alone. In any case, the person skilled in the art will understand that such graphical representations of data may be subject to small variations, for example, in relative peak intensities and peak positions due to certain factors such as, but not limited to, variations in instrument response and variations in sample concentration and purity, which are well known to experts. However, the skilled person will readily be able to compare the graphical data in the Figures herein with graphical data generated for an unknown solid / crystalline form and confirm whether the two graphical data sets characterize the same solid / crystalline form. crystalline or two solid / crystalline state forms. A solid / crystalline state form of a Sofosbuvir referred to herein as being characterized by graphical data (as illustrated in "or" as substantially illustrated in "a Figure, will thus be understood to include any form Sofosbuvir solid / crystalline state characterized with graphical data having such small variations, as is well known to experts, compared to Figure.

As used herein and unless otherwise indicated, the term "anhydride" with respect to crystalline / solid state forms of Sofosbuvir, refers to a crystalline / solid state form of Sofosbuvir that does not include water (or other solvents) in a defined, stoichiometric amount within the glass. On the other hand, an "anhydrous" form would typically contain no more than 1% (w / w) of neither water nor organic solvents as measured, for example, by TGA.

As used herein, the term "isolated" with respect to solid state forms of Sofosbuvir of the present invention corresponds to a solid state form of Sofosbuvir that is physically separated from the reaction mixture in which it is formed. .

As used herein, unless otherwise noted, DPRX measurements are taken using 1.5418A wavelength copper Ka radiation. Unless otherwise stated, 2-theta DPRX values are reported with an error of 0.2 degrees 2-theta.

The individual particles of a sample or aliquot of the solid particles comprising the crystalline form E of Sofosbuvir of the present invention are not uniform in size. Instead, a sample or aliquot of a Solid particle comprising crystalline form E of Sofosbuvir of the present invention is comprised of different sizes which can be sized or distributed in a grouping of distinct adjacent ranges of particle size. If the size of the intervals is small enough, the grouping of particle sizes approaches a continuum of particle sizes. This collection of discrete particle size ranges along with their population is referred to as the particle size distribution (DTP).

The measurement and characterization of particle size distributions is known in the art. It is possible to compare particle samples comprising the crystalline E form of Sofosbuvir based on individual points on a cumulative particle size distribution curve. Measurements are represented as d (0, X) = Y (where X and are Arabic numerals), each "d" describes an individual point on a cumulative DPT curve. The number "X" represents the percentage (number, volume, or weight) of particles in the population that have a nominal size up to and that include "Y". Hence, d (0.9) = 250 p is characteristic of a DPT in which 90% (number, volume, or weight) of the particles in a population have a nominal size of about 250 p mense (at least some particles having a nominal dimension of 250 p), and so on. When the DPT is determined by the well-known laser diffraction method described herein, the d (0, X) measurement illustrates a volume average.

It is known to the person skilled in the art that the results of DPT determination by one technique can be correlated with results of another technique on an empirical basis by routine experimentation.

As used herein, unless otherwise indicated, "median particle size" refers to the D50 value of the particle size distribution. As used herein, the particle size distribution is determined by means of laser diffractometry. More specifically, unless otherwise noted, particle size was determined using a Malvern Instruments Mastersizer 2000 - particle size determination can be carried out as a wet or dry measurement depending on the sample.

Median particle size (D50), which is also denoted D50 value of the integral volume distribution, is defined in the context of the present invention as the particle diameter, where 50 volume percent of particles have a diameter less than the diameter that corresponds to the D50 value. Also, 50 percent of the volume of the particles has a diameter greater than the D50 value. Similarly, the D90 value of the integral volume distribution is defined as the particle diameter, where 90 volume percent of particles have a diameter less than the diameter, which corresponds to the D90 value. Correspondingly, the D10 value of the integral volume distribution is defined as the particle diameter, where 10 volume percent of particles have a diameter less than the diameter, which corresponds to the D10 value.

According to the invention, "particle diameter" or "particle size" of a particle to be determined refers to the diameter of an equivalent particle that is assumed to be spherical and has the same light scattering pattern as the particle a decide.

An object, eg, a reaction mixture may be characterized herein as being at, or allowed to return to, "room temperature" or "room temperature", unless abbreviated as "RT". This means that the temperature of the object is close to, or is the same as, that of the space, for example the room or fume hood, in which the object is located. Generally, the ambient temperature is from about 20 ° C to about 30 ° C, or about 22 ° C to about 27 ° C or about 25 ° C.

The amount of solvent employed in a chemical process, for example a reaction or a crystallization, may be referred to herein as the number of "volumes" or "vol" or "V". For example, a material can be referred to as being suspended in 10 volumes (or 10 vol or 10V) of a solvent. In this context, this term would be understood to refer to milliliters of the solvent per gram of the material being suspended, so that suspending 5 grams of a material in 10 volumes of a solvent means that the solvent is used in an amount 10 milliliters of the solvent per gram of the material being suspended, in this example 50 ml of the solvent. In another context, the term "v / v" can be used to indicate the number of volumes of a solvent that are added to a liquid mixture based on the volume of that mixture. For example, the addition of solvent X (1.5 v / v) to 100 ml of reaction mixture would indicate that 150 ml of solvent X was added.

A process or step may be referred to herein as taking place "overnight". This refers to the time interval, for example, for the process or stage, which elapses in time during the night, when a process or stage cannot be actively observed. This time interval is from about 8 to about 20 hours, or about 10-18 hours, typically about 16 hours.

As used herein, the term "reduced pressure" refers to a pressure that is less than atmospheric pressure. For example, the reduced pressure is from about 10 mbar to about 50 mbar. As used herein, unless otherwise indicated, the term "vacuum" refers to a pressure of from about 0.2 mbar to about 10 mbar, from about 0.2 to about 5 mbar, about 0.5 to about 4 mbar, about 1 mbar to about 3 mbar, and preferably about 2 mbar.

As used herein, an "antisolvent" is a liquid that when combined with a composition comprising a solvent or sofosbuvir, induces precipitation of crystalline Sofosbuvir.

As used herein, unless otherwise indicated, the term "compressibility" is expressed as maximum density at constant pressure of 0.2 MPa.

As used herein, Sofosbuvir crystalline form 1 refers to a crystalline form that can be characterized by X-ray powder diffraction pattern as illustrated in Figure 2. Sofosbuvir crystalline form 1 can be characterized by DpRX peaks from the X-ray powder diffractogram disclosed in US 2011/0251152 or WO2011 / 123645. As used herein, Sofosbuvir form 6 refers to a crystalline form as described in WO 2011/0251152, which can be characterized by X-ray powder diffraction pattern as illustrated in Figure 3 Thus, crystalline Sofosbuvir Form 6 can be characterized by an X-ray powder diffraction pattern having peaks as disclosed in US 2011/0251152 or WO2011 / 123645, which is depicted in Figure 3.

The crystalline forms of Sofosbuvir prepared by one described herein may be substantially free of any other form of Sofosbuvir or the specified polymorphic forms of Sofosbuvir, respectively.

The present invention comprises a process for the preparation of the aforementioned pharmaceutical compositions. The process comprises combining the solid state Sofosbuvir E form with at least one pharmaceutically acceptable excipient.

In one embodiment, the present invention comprises a crystalline form of Sofosbuvir, designated Form E, characterized by data selected from one or more of the following: an X-ray powder diffraction pattern as illustrated in Figure 1 or Figure 7 ; an X-ray powder diffraction pattern having peaks at: 12.4, 16.2, 17.2, 25.0 and 25.3 degrees two theta ± 0.1 degrees two theta; an X-ray powder diffraction pattern having peaks at: 12.4, 16.2, 17.2, 25.0 and 25.3 degrees two theta ± 0.1 degrees two theta and no peaks at: 10 , 9 and 14.2 degrees two theta ± 0.2 degrees two theta; and combinations of these data.

The crystalline E form of Sofosbuvir can be further characterized by data selected from one or more of the following: X-ray powder diffraction pattern having peaks at 12.4, 16.2, 17.2, 25.0, and 25 , 3 degrees two theta ± 0.1 degrees two theta and also one, two, three or four peaks selected from: 8.1, 19.4, 22.0 and 23.3 degrees two theta ± 0.1 degrees two theta ; an X-ray powder diffraction pattern having peaks at: 12.4, 16.2, 17.2, 25.0 and 25.3 degrees two theta ± 0.1 degrees two theta, no peaks at: 10 , 9 and 14.2 degrees two theta ± 0.2 degrees two theta and also having one, two, three or four peaks selected from: 8.1, 19.4, 22.0 and 23.3 degrees two theta ± 0, 1 degrees two theta.

The crystalline E form of Sofosbuvir can be characterized by an X-ray powder diffraction pattern having peaks at 8.1, 12.4, 16.2, 17.2, 19.4, 22.0, 23.3, 25.0 and 25.3 degrees two theta ± 0.1 degrees two theta. Alternatively, the crystalline E form of Sofosbuvir can be characterized by an X-ray powder diffraction pattern that has peaks at 8.1, 12.4, 16.2, 17.2, 19.4, 22.0, 23 , 3, 25.0 and 25.3 degrees two theta ± 0.1 degrees two theta and an absence of peaks at 10.9 and 14.2 degrees two theta ± 0.2 degrees two theta.

The crystalline E form of Sofosbuvir can be further characterized by the data indicated in the table below. Table: Os X peaks of Form E Sofosbuvir:

______ (continuation)_____

Angle (degrees two theta ±

0.1 degrees two theta ._____

22.0 ___________________

23.3 __________________

23.7 ___________________

24.4 __________________

In another embodiment, the crystalline form E can be characterized by X-ray powder diffraction pattern having peaks at: 8.1, 12.1, 12.4, 13.5, 16.2, and 17.2 degrees two theta. ± 0.1 degrees two theta. In a preferred embodiment, the present invention relates to a pharmaceutical composition comprising the E form of sofosbuvir, wherein the E form is characterized by an X-ray powder diffraction pattern having peaks at: 8.1, 12.1 , 12.4, 13.5, 16.2 and 17.2 degrees two theta ± 0.1 degrees two theta.

In another embodiment, Form E can be characterized by X-ray powder diffraction pattern having peaks at: 8.1, 12.1, 12.4, 13.5, 16.2, and 17.2 degrees two theta ± 0.2 degrees two theta and no peaks at: 10.9 and 14.2 degrees two theta ± 0.2 degrees two theta. In a preferred embodiment, the present invention relates to a pharmaceutical composition comprising Form E of sofosbuvir wherein Form E is characterized by an X-ray diffraction pattern having peaks at 8.1, 12.1, 12, 4, 13.5, 16.2 and 17.2 degrees two theta ± 0.2 degrees two theta and no peaks at: 10.9 and 14.2 degrees two theta ± 0.2 degrees two theta.

In another embodiment, the crystalline form E can be characterized by X-ray powder diffraction pattern having peaks at: 8.1, 10.4, 13.5, 18.0, and 18.7 degrees two theta ± 0.1 degrees two theta. In a preferred embodiment, the present invention relates to a pharmaceutical composition comprising the E form of sofosbuvir, wherein the E form is characterized by an X-ray powder diffraction pattern having peaks at: 8.1, 10.4 , 13.5, 18.0 and 18.7 degrees two theta ± 0.1 degrees two theta.

In another embodiment, Form E can be characterized by X-ray powder diffraction pattern having peaks at: 8.1, 10.4, 13.5, 18.0, and 18.7 degrees two theta ± 0.2 degrees. two theta and absence of peaks at: 10.9 and 14.2 degrees two theta ± 0.2 degrees two theta. In a preferred embodiment, the present invention relates to a pharmaceutical composition comprising Form E of sofosbuvir wherein Form E is characterized by an X-ray diffraction pattern having peaks at 8.1, 10.4, 13, 5, 18.0 and 18.7 degrees two theta ± 0.2 degrees two theta and no peaks at: 10.9 and 14.2 degrees two theta ± 0.2 degrees two theta.

Form E according to the invention can be a monoclinic crystalline form of sofosbuvir, space group P21 having the following unit cell parameters -14.47 A, b -17.02 A, c -5, 22 A, alpha ~ 90.00 °, beta ~ 100.30 ° and gamma ~ 90.00 °, preferably as obtained by synchrotron radiation at a wavelength of 0.40003 (1) A. It can be understood that the above parameters of Unit cell can be obtained by indexing a high resolution powder pattern obtained in synchrotron radiation at a wavelength of 0.40003 (1) A.

In one embodiment, Form E is anhydrous.

Form E has advantages as described above. In particular, the E form is stable for at least 3 months at 40 degrees and 25 degrees.

In one embodiment of the present invention, the crystalline form E of Sofosbuvir is isolated.

Compositions comprising the crystalline E form of the present invention are preferably polymorphically pure, that is, substantially free of any other form of sofosbuvir. For example, the crystalline E form of the present invention is substantially free of sofosbuvir form 6. Specifically, the crystalline E form of the present invention contains 20% or less, 10% or less, 5% or less, 2% or less, 1% or less, of the form 6 sofosbuvir measured by DRXP. Accordingly, the content of crystalline Sofosbuvir form 6 in crystalline Sofosbuvir form E will be measured by detecting and quantifying the characteristic peaks described in form 6. The characteristic peaks of crystalline Sofosbuvir form 6 used for the measurement described above can be either one or both peaks at: 6.08 and 10.85 degrees two theta ± 0.2 degrees two theta. In a preferred aspect, the present invention encompasses solid forms of Sofosbuvir comprising the crystalline E form of Sofosbuvir which has preferred hygroscopic properties, ie, it is less hygroscopic compared to Form 1 and Form 6 of Sofosbuvir. This property can be demonstrated by the DVS data presented in Figures 4, 5 and 6 and by the following table:

Table 2

continuation

Solid forms of crystalline form E with low hygroscopicity, according to the present invention, can be easily handled, processed and stored, without the need for special controlled conditions and can therefore be used to advantage for the preparation of compositions pharmaceuticals and formulations.

The improved hygroscopic properties described herein above can be expressed by the dynamic water absorption controlled by the DVS instrument. The dynamic curve profile obtained can provide insight into the substance's tendency to deliquescence. This is expressed by a sudden change in the slope of the curve (inflection point) that indicates that at a certain critical relative humidity the substance begins to absorb significant amounts of water. This behavior indicates a long-term possibility of the stored powder to stick and form lumps.

The DVS curve of sofosbuvir form 1 (illustrated in Figure 4) shows a critical relative humidity in the sorption curve at approximately 80% RH at which rate the water absorption changes (inflection point). Significant water absorption is observed above 80% RH (approximately 5%). This indicates that the sample may partially liquefy during long-term storage or during exposure to high humidity.

The DVS curve of form 6 (illustrated in Fig. 5) of sofosbuvir shows a critical relative humidity in the sorption curve at approximately 70% RH at which rate the water absorption changes (inflection point). Significant water absorption is observed above 70% RH (approximately 1%). This indicates that this sample may also partially liquefy during long-term storage or during exposure to high humidity.

It was found, surprisingly, that Sofosbuvir comprising the low hygroscopicity Form E does not show this tipping point. Furthermore, this E form does not show any hysteresis between desorption and isothermal absorption. The hysteresis between absorption and desorption is defined as the loop that appears in the figure as a result of the gap that forms between the absorption weight increase value and the desorption weight decrease value at a given relative humidity. A comparison of the water content values as measured by DVS is shown in Table 2 above.

The crystalline E form of sofosbuvir can be characterized by a water vapor sorption isothermal showing a water absorption of not more than 0.15-0.25% by weight, preferably not more than 0.15% by weight, at a relative humidity of up to 90%. The crystalline E form of sofosbuvir can be characterized by a water vapor sorption isothermal as shown substantially in Figure 6.

The crystalline E form of sofosbuvir can be characterized by a water vapor sorption isothermal showing a water absorption of no more than 0.10% by weight, at a relative humidity (RH) of up to 70% and / or a water absorption of not more than 0.15-0.25% by weight, preferably not more than 0.15% by weight, at a relative humidity of up to 90%.

The crystalline E form of Sofosbuvir of the invention can be characterized by a water vapor sorption isothermal showing a water absorption of not more than 0.15-0.25%, preferably not more than 0.15%, at a relative humidity of up to 90%.

The crystalline E form of Sofosbuvir of the invention can be characterized by a water vapor sorption isothermal showing a water absorption of no more than 0.10% by weight, at a relative humidity (RH) of up to 70% and / or a water absorption of not more than 0.15-0.25% by weight, preferably not more than 0.15% by weight, at a relative humidity of up to 90%.

The crystalline E form of Sofosbuvir may have a low hygrocospicity, characterized by a water vapor sorption isothermal showing a water absorption of not more than 0.15-0.25% by weight, preferably not more than 0.15 % by weight, at a relative humidity of up to 90%.

The crystalline E form of Sofosbuvir may have low hygroscopicity characterized by a water vapor sorption isothermal showing a water absorption of not more than 0.10% by weight, a relative humidity (RH) of up to 70% and / or a water absorption of not more than 0.15-0.25% by weight, preferably not more than 0.15% by weight, at a relative humidity of up to 90%.

The crystalline form E of sofosbuvir can be prepared by a process comprising (a) combining sofosbuvir with a solvent system comprising an organic solvent and optionally water (b) optionally seeding with crystals of Form E (c) adding an antisolvent (d ) optionally isolating the crystalline E form and (e) grinding or optionally micronizing to obtain sofosbuvir crystalline form E having a lower particle size. The process can also comprise filtering and drying stages.

Preferred organic solvents in step (a) can be selected from a group consisting of aliphatic ethers, preferably C4-C8 ethers, cyclic ethers, ketones, preferably C3-C8 ketones, alcohols, preferably C4-C8 alcohols. More preferably, the organic solvents can be selected from a group consisting of methyl t-butyl ether, t-butanol, acetone, methyl isobutyl ketone. Most preferably, the organic solvent is methyl isobutyl ketone.

The preferred solvent used as antisolvent in step (c) can be selected from a group consisting of aliphatic ethers, preferably C4-C8 ethers, alkanes, preferably C3-C8 alkanes and cycloalkanes. More preferably, the antisolvents can be selected from a group consisting of di-isopropyl ether, n-heptane, cyclohexane. Most preferably, the antisolvent is cyclohexane.

Step (a) is preferably carried out in a solvent system comprising an organic solvent and water. More preferably, the amount of water can be in the range of 0% -2% by volume.

Preferably, the reaction mixture from step (a) includes from about 10% to about 50% Sofosbuvir, by volume. Preferably, the process is carried out at temperatures between 0 to 20 ° C, more preferably between 0-15 ° C, most preferably 6-12 ° C.

The solution obtained in step (a) can be filtered, if desired, to get rid of foreign particles, while keeping the filtered solution and the filtrate at almost the same temperature. Preferably, the process is carried out with agitation

Generally, the amount of crystalline form E of Sofosbuvir used for seeding in step (b) is about 0.1% to 0.5% by weight of the Sofosbuvir.

The crystalline E form of Sofosbuvir can be isolated by any method known in the art. For example, the crystalline E form of Sofosbuvir can be separated by filtering the suspension or decanting the solvent from the suspension. The isolation method may further comprise washing and drying the crystalline form E of Sofosbuvir. Preferably, the crystalline form E of Sofosbuvir is dried at a temperature of about 30 ° C to about 60 ° C, more preferably, at a temperature of about 40 ° C to about 50 ° C under reduced pressure.

The crystalline E form of Sofosbuvir can be ground or micronized to obtain sofosbuvir having a smaller particle size in a process that is tailored to the desired particle size. For example, micronization can be carried out using a 100mm plate air jet micronizer.

Another process to prepare the crystalline form E of Sofosbuvir comprising: a) combining Sofosbuvir with methyl isobutyl ketone and water b) seeding with crystals of form E, c) adding cyclohexane d) optionally isolating form E and (e) grinding or micronizing optionally to obtain the crystalline form E of Sofosbuvir having a desired particle size. The process can also comprise filtering and drying stages.

The starting material used in the process described above can be Sofosbuvir crystalline form 1. The amount of water in step a) can be in the range of 0.5 to 2% (by volume) and the reaction mixture is stirred. Preferably, the amount of water is 1% (by volume) and the reaction is stirred at 50-200 RPM. The reaction mixture can be further filtered.

Stage a) can be carried out at a temperature ranging from 0 to 20 ° C, preferably, stage a) is carried out at 9 ± 3 ° C.

A preferred process to prepare the crystalline form E of Sofosbuvir comprises: a) combining Sofosbuvir with methyl isobutyl ketone and water where the amount of water is 1% by volume and the temperature is 9 ± 3 ° C b) seeding with crystals form E, c) add cyclohexane d) isolate form E and (e) optionally micronize. The process can also comprise filtering and drying stages.

Step a) can be carried out with stirring at a speed of 50-200 RPM. Preferably, the solution is stirred after seeding, the stirring speed is in the range of 50-200 RPM.

Most preferably, sofosbuvir crystalline form 1 is combined with MIBK and water (1% by volume) at 9 ± 3 ° C. The resulting suspension is stirred at 50-200 RPM until the solution becomes clear. The solution is filtered to remove any foreign particles. The filtered solution is charged, stirred at at 9 ± 3 ° C, seeded with crystals of form E and stirred as such for 4 ± 0.5 h. Cyclohexane is then added and the solution is stirred for 1 ± 0.5 h at 9 ± 3 ° C. The suspension is filtered and the cake is washed with cyclohexane. The solid obtained was dried under vacuum for 12 h at Tj 40-50 ° C to obtain Sofosbuvir Form E.

As disclosed herein, the micronized or small particle size E Cristina form of sofosbuvir having a desired DTP is suitable for the preparation of pharmaceutical compositions and formulation, for example, d (0.1) 0.4- 0.6 microns, d (0.5) 1.0-2.0 microns and d (0.9) 3.2-5.0 microns.

Small particle size sofosbuvir can be directly obtainable by different processes, for example, it can be obtained by chemical synthesis or by micronization techniques, optionally, by using deagglomeration techniques.

Micronization is a technique known to those of skill in the art. For example, by grinding, as a commonly known micronization technique, small particle sizes can be achieved.

It is generally known in the art that micronized or small particle size material can lead to downstream processing problems related to poor flow and dispersion properties. It was surprisingly found that the micronized or small particle size Sofosbuvir comprising crystalline form E of the present invention shows good flowability and compressibility and reduced hygroscopicity, therefore, it can be advantageously used for the preparation of pharmaceutical compositions and formulations.

The physical attributes of PFA are especially important for the reproducibility of dosage form preparations with a high content of PFA. A high drug load may have flow problems (http://en.wikipedia.org/wiki/Power_flow "\ or" Power flow) or require large dosage forms if the compound has low compressibility. Therefore, the crystalline form E of micronized or small particle size Sofosbuvir is suitable for use in the preparation of formulations containing a high loading of sofosbuvir, for example containing more than 35% sofosbuvir. In addition, sofosbuvir micronized or small particle size crystalline Sofosbuvir form E is particularly suitable for use in the preparation of dry formulations containing a high dose of sofosbuvir as a single PFA or in combination with another pharmaceutically active ingredient, for example, ledipasvir.

Micronized or small particle size Sofosbuvir comprising crystalline Form E of Sofosbuvir may have preferential flow properties, that is, it exhibits a flow rate of only 100 s / 100 g, more preferably no more than 80 s / 100 g, plus preferably no more than 65 s / 100 g, as measured by flow into a hole in the Pharma-Test 41-03300 automated powder assay system.

Sofosbuvir crystalline E-form micronized or small particle size Sofosbuvir may have advantageous compressibility properties, i.e. showing a constant density of not less than 0.40 g / ml, preferably not less than 0.43 g / ml, more preferably not less than 0.48 g / ml as measured at 0.2 MPa.

Sofosbuvir crystalline E-form micronized or small particle size Sofosbuvir can be characterized by a water sorption of not more than 0.5%, more preferably not more than 0.3% as measured by DVS and water content after equilibration at 90% RH.

The sofosbuvir micronized or small particle size crystalline E form of sofosbuvir can be characterized by any one of the above aspects or any combination thereof.

The micronized or small particle size crystalline form E can be characterized by DTP of d (0.1) 0.4-0.6 microns, d (0.5) 1.0-2.0 microns and d (0, 9) 3.2-5.0 microns; a flow rate of no more than 100 s / 100 g, more preferred no more than 80 s / 100 g, more preferred no more than 65 s / 100 g; a constant density not less than 0.40 g / ml, preferably not less than 0.48 g / ml as measured at 0.2 MPa; and a water sorption of not more than 0.5%, more preferably not more than 0.3% as measured by DVS and water content after equilibration at 90% RH.

The micronized or small particle size crystalline form E can be characterized by DTP of d (0.1) 0.4-0.6 microns, d (0.5) 1.0-2.0 microns and d (0, 9) 3.2-5.0 microns; a flow rate of no more than 65 s / 100 g; a constant density not less than 0.48 g / ml as measured at 0.2 MPa; and a water sorption of more than 0.3% as measured by DVS and water content after equilibration at 90% RH.

The previously described micronized or small particle size form of Sofosbuvir can be used to prepare pharmaceutical compositions and formulations.

The present invention encompasses pharmaceutical compositions and formulations comprising the crystalline Sofosbuvir form E of the present invention, in particular, formulations containing a high loading of sofosbuvir, eg, containing more than 35% sofosbuvir. The pharmaceutical composition is generally a solid composition and Sofosbuvir retains its solid form. The pharmaceutical composition may comprise sofosbuvir as a PFA alone or in combination with another pharmaceutically active ingredient, for example ledipasvir. Pharmaceutical compositions can be prepared by a process that comprises combining the form Sofosbuvir crystalline of the present invention with at least one pharmaceutically acceptable excipient.

The above crystalline Sofosbuvir form E of the present invention can also be used as medicaments. The present invention further encompasses 1) the use of the aforementioned crystalline form of Sofosbuvir in the manufacture of a pharmaceutical composition and 2) a pharmaceutical composition comprising the crystalline E form of Sofosbuvir described herein for use in the treatment of Hepatitis C.

Solid pharmaceutical compositions according to the present invention are, for example, powders, granules, agglomerates, capsules or tablets.

The pharmaceutical composition of the present invention is preferably in the form of a tablet comprising one or more pharmaceutically acceptable excipients. The tablet for oral use.

Pharmaceutical compositions in the form of a tablet according to the present invention are preferably for use in treating hepatitis C infections as is known in the art. As indicated above, in such use the compositions in the form of a tablet according to the present invention can be used alone or in the form of a combination therapy as is known in the art.

Processes for the preparation of pharmaceutical compositions in the form of a tablet according to the invention are known in the art. Sofosbuvir can be dry granulated or wet granulated prior to tabletting or mixtures of tablets can be directly compressed. Granulation is preferred for the preparation of sofosbuvir tablets and dry granulation is most preferred. Tablets according to the invention can be coated to facilitate their intake and / or to enhance the appearance of the tablet. In a preferred embodiment the pharmaceutical compositions in the form of a tablet according to the present invention are prepared by dry granulation. It is preferred that in the dry granulation method, a tablet is first formed, then the tablet is crushed to obtain particles that are easier to handle than the original mixture. The crushed particles are screened, optionally mixed in a mixer such as a rotary motion mixer, and then pressed into the pharmaceutical composition in the form of a tablet as described herein.

Preferably, the pharmaceutical compositions according to the present invention comprise small particle size sofosbuvir Form E. In another embodiment, the pharmaceutical composition comprises small particle size sofosbuvir Form E without glidants.

Glidants are known to experts. They reduce interparticle friction and cohesion and thus increase the fluidity of particles or mixtures. Colloidal silicon dioxide is the best known glidant. Unlike glidants, lubricants prevent ingredients from sticking, for example, to perforations in the tablet. Magnesium stearate is a typical lubricant (not a glidant).

The composition comprising small particle size sofosbuvir preferably comprises no more than 1.5% lubricants based on the weight of the total composition. In particular, the composition of the present invention preferably comprises not more than 1.5% magnesium stearate, more preferably not more than 1%, most preferably not more than 0.6% magnesium stearate based on the weight of the total composition. For pharmaceutical compositions comprising glidant-free small particle size sofosbuvir, the particle size distribution is measured by laser diffraction and is preferably referred to a volume distribution. Preferably, the particle size distribution is measured using the Malvern Mastersizer 2000. Small particle size sofosbuvir preferably has a d (0.5) of less than 20 pm, preferably less than 10 pm, more preferably less than 5 pm, and most preferably between 1-2 pm. In one embodiment, the small particle size sofosbuvir has a d (0.1) of 0.4-0.6 pm, a d (0.5) of 1.0-2.0 pm, and a d (0 , 9) from 3.2-5.0 pm. The values of d (0.1), d (0.5), and d (0.9) are based on a volume distribution. For the measurement of sofosbuvir particle size reference is also made to Example 22.

The following Examples are set forth to aid in an understanding of the invention. Examples marked with an asterisk (*) are provided for reference only.

Methods

X-ray powder diffraction (Examples 1-13)

X-ray powder diffraction analyzes were carried out on a Bruker D8 Advance X-ray powder diffractometer; CuKa radiation (A = 1.5418 A); lynx eye detector; 22-25 ° C laboratory temperature; PMMA specimen support ring. Before analysis, the samples were gently ground by means of a mortar and pestle to obtain a fine powder. The crushed sample was fitted in a cavity of the same holder and the sample surface was smoothed by means of a glass cover.

Measurement parameters:

Scan range: 2-40 degrees 2-theta;

Scan mode: continuous;

Stage size: 0.05 degree;

Time per stage: 0.5 s;

Sample rotation: 30 rpm;

Sample holder: PMMA specimen holder ring.

The peak positions described were determined using silicon powder as an internal standard in a mixture with the measured sample. The position of the silicon (Si) peak was corrected to the theoretical peak of the silicone: 28.45 degrees two-theta and the positions of the measured peaks were corrected respectively.

DVS measurements

DVS measurements were performed according to SMS of the analysis parameters of DVS instrument Analysis:

Mass: 20-30 mg

Temperature: 25 ° C

Humidity range: 0% -90% -0% cycle.

Stage size: 10% RH

dm / dt: 0.002% / min

minimum stage duration: 60 min

maximum stage duration: 720 min

Fluency

The flow rate (s / 100 g powder) was measured on a Pharma-Test 41-03300 flow meter. The apparatus consists of a conical stainless steel powder funnel on top of the apparatus equipped with an automatic stirring paddle and a 15mm pouring nozzle below the funnel. The method consists of pouring a quantity of powder with a stirring regime through a given size nozzle and weighing the weight of the powder that has passed through the nozzle and the time it has taken. Results are given as s / 100g.

DTP Measurement Method of Form 7

If a wet measurement is used the particles to be measured are dispersed in a dispersant, preferably having a refractive index (IR) of 1.403, and the measurement is carried out at 2000 rpm and ultrasound for 120 s before the measurement. Preferably, unless otherwise indicated, the particle size distribution is determined using a wet measurement using a Malvern Instruments Mastersizer 2000 and Malvern 2000S wet sample of the dispersion unit and silicone oil as dispersant. Unless otherwise indicated, the following configurations are preferably used:

DTP Measurement Method of Form E-Dry Cell Measurement

Mastersizer 2000

Analysis model: General object

Dust IR: 1.573

Absorption: 0.5

Dispersant name: Dry dispersion (air)

Dispersant IR: 1.0

Sensitivity: Normal (fine powder)

Filtering Darkening: On (0.5-3%)

Measurement time: 5 seconds (5000 snapshots)

Background measurement time: 5 seconds (5000 snapshots)

Aliquot repeats: 1

Repeat measurement: 1

Pressure: 2.5 Bar Flow cell feed rate: 30%

Funnel gap: 5mm

Small sieve feeder filled with balls (2/3 by volume)

Between sample runs, clean the cell with air for 30 seconds.

A background measurement should be performed before each measurement. Add approximately 0.5 g of sofosbuvir powder with a spatula directly into the feeder. Start the feeder job. Darkening should be in the range of 0.5-3%.

Examples

For Examples 1-13, Sofosbuvir was prepared according to the method described in B. S. Ross, P. G. Reddy, H.-R. Zhang, S. Rachakonda, M. J. Sofia, J. Org. Chem. 2011, 76, 8311. Sofosbuvir Form 1 can be obtained according to the process described in WO 2010/135569.

Example 1: Preparation of crystalline form E of Sofosbuvir

Sofosbuvir (1 g, Form 1) was dissolved in acetone (3 ml) with stirring at 25 ± 5 ° C. Then, di-iso-propyl ether (DIPE) (10 ml) was added with stirring to make turbidity. The mass was stirred for 2 h at 25 ± 5 ° C to provide a white solid. DIPE (15 ml) was added to this mass and the mass was stirred for 48 h at 25 ± 5 ° C. The mass was filtered and the obtained solid was dried at 50 ° C under vacuum for 12 h to provide Sofosbuvir E form (as confirmed by DPRX).

Example 2: Preparation of crystalline form E of Sofosbuvir

Methyl iso-butyl ketone (MIBK) (3 ml) was charged in 100 ml into a round bottom glass flask equipped with a mechanical stirrer. This was cooled to 0 to 5 ° C with stirring and Sofosbuvir (1 g, Form-1) was added at 0 to 5 ° C. This mass slowly turned clear after 15 min at 0 to 5 ° C. At this point, Seed Sofosbuvir Form E (10 mg) were added at the same temperature. After 10 min of stirring at the same temperature, the formation of a precipitate was observed. The mass was stirred for 1 hr at 0 to 5 ° C. Cyclohexane (10 ml, precooled to 15 ° C) was added and stirred for 2 to 5 min. The precipitate was filtered and washed with cyclohexane (10 ml, pre-cooled to 15 ° C). The solid was dried at 33 ° C under vacuum for 12 h to provide Sofosbuvir form E (as confirmed by DPRX).

Example 3: Preparation of crystalline form E of Sofosbuvir

(MIBK) (60 ml) in 250 ml was loaded into a round bottom glass flask equipped with a mechanical stirrer. This was cooled to 10-15 ° C with stirring and Sofosbuvir (20g, Form 1) was added at 10-15 ° C. This mass slowly turned clear after 20 min at 10 to 15 ° C. At this point, seeded Sofosbuvir Form E (100 mg) were added at the same temperature. After 10 min of stirring at the same temperature, the formation of a precipitate was observed. The mass was stirred for 1 hr at 10-15 ° C. Cyclohexane (60 ml, precooled to 15 ° C) was added slowly and stirred for 2 to 5 min. The precipitate was filtered and washed with cyclohexane (60 ml, pre-cooled to 15 ° C). The obtained solid was dried at 33 ° C under vacuum for 12 h, to provide Sofosbuvir form E (as confirmed by DPRX).

Example 4: Preparation of crystalline form E of Sofosbuvir

(MIBK) (3 ml) in 100 ml was loaded into a round bottom glass flask equipped with a mechanical stirrer. This was cooled to 10-15 ° C with stirring and Sofosbuvir (1g, Form 1) was added at 10-15 ° C. This mass slowly turned clear after 15 min at 10 to 15 ° C. At this point, Seed Sofosbuvir Form E (10 mg) were added at the same temperature. After 10 min of stirring at the same temperature, the formation of a precipitate was observed. The mass obtained was stirred for 1 h at 10-15 ° C. Cyclohexane (10 ml, precooled to 15 ° C) was added and stirred for 2 to 5 min. The precipitate obtained was filtered and washed with cyclohexane (10 ml, pre-cooled to 15 ° C). The solid was dried at 33 ° C under vacuum for 12 h to provide Sofosbuvir form E (as confirmed by DPRX).

Example 5: Form E single crystal X-ray crystallography

Sofosbuvir Form E was sealed in 1 mm capillaries (1257-67A), No. 50 glass. The powder diffraction pattern was measured using synchrotron radiation at a wavelength of 0.40003 (1) A. they determined the mirror positions in DASH software and indexing was performed in DICVOL91 software. The structure was dissolved using the DASH 3.2 program. The powder structure refinement was performed in JANA2006 software. The presence of any solvent was not detected.

Crystal lattice parameters of Sofosbuvir, Form E:

cell_length_a 14.4723 (2) A

cell_length_b 17.0212 (2) A

cell_length_c 5.22862 (6) A

alpha_cell_angle 90.0000 °

beta_cell_angle 100.297 (1) °

gamma_cell_angle 90.0000 °

volume-cell 1267.3 A3

symmetry_space_group_name_: P21

monoclinic_cell_symmetry configuration

cell_measuring_temperature 293 K

Example 6: Preparation of crystalline form E of Sofosbuvir

Sofosbuvir Form 1 (100 g) along with MIBK (500 ml) was charged into a 4 neck round bottom flask equipped with a mechanical stirrer. The moisture content of the reaction was adjusted to 0.5-2.0% by volume and the material was dissolved with stirring at 10 + 5 ° C. Sofosbuvir Form E crystals (1 g) were added to the reaction matrix and stirring was continued at the same temperature. During this interval, a white solid slowly precipitated out of the reaction mixture. After 4-7 hours, cyclohexane (1000 ml) was added and the resulting suspension was filtered. The obtained white solid was washed with cyclohecane (2 X 200 ml) and sexed at 45 ° C under vacuum for 12 h to provide a Sofosbuvir Form E (as confirmed by DPRX).

Example 7: Preparation of crystalline form E of sofosbuvir

MIBK (5 L) and water (1% MIBK) were added to Sofosbuvir Form-1 (1 Kg) at 9 ± 3 ° C. The resulting suspension was stirred at 50-200 RPM until the solution became clear. The solution was filtered to remove any foreign particles. It was washed with MIBK (0.5 L). The filtered solution was charged, stirred at at 9 ± 3 ° C, seeded with Form E crystals and the solution was stirred as such for 4 ± 0.5 h at 50-200 RPM. Then cyclohexane (10 L) was added and the solution was stirred for 1 ± 0.5 h at 9 ± 3 ° C. The suspension was filtered and the cake was washed with cyclohexane (2 X 2 L). The solid was dried under vacuum for 12 h at Tj 40-50 ° C to obtain Form E in 80 to 90% yield.

Example 8: Preparation of micronized Sofosbuvir crystalline form E

Micronized Sofosbuvir crystalline form E was prepared using a 100mm plate air jet micronizer, feed rate 0.5 Kg / h, feed pressure 6.5 bar +/- 0.5 bar, grind pressure 6 bar +/- 0.5 bar.

Example 9: Fluidity of micronized sofosbuvir crystalline E-form

70 g of the micronized crystalline Form E powder of Sofosbuvir obtained according to Example 8 was poured into the funnel. Stirring was started at 20 rpm, the pouring nozzle was opened and the powder began to flow through the nozzle. After approximately 30 seconds the flow stopped. The exact weight of the powder that passed through the hole and the exact time in seconds were automatically recorded. The resulting flow rate, measured in duplicate analysis, was: 63 s / 100 g and 62 s / 100 mg.

Example 10: Crystalline Form E compressibility of micronized sofosbuvir

Compression profiles were obtained using a flat face punch and punch set on a TA-XT2 texture analyzer (Stable Micro Systems Ltd., Godalming, UK).

200 mg of the crystalline form E of micronized Sofosbuvir obtained according to Example 8 is compressed in an aceto mold (with the displacement rate of 0.03 mm / s). A cyclic procedure (similar to threading) was carried out: compress, then refract, relax for 15 s and then repetitive compressive steps (total 10 steps). Each stage exerts a pressure of 0.2 MPa on the sample. The density of the sample is calculated by dividing the weight by the volume of the sample for each cycle. The maximum density is reached within 10 stages. The maximum compressive displacement was 0.5 mm. The resulting density was: 0.48 g / ml

Example 11: Preparation of crystalline form E of Sofosbuvir

Sofosbuvir (Form 1, 1 g) was suspended in methyl tert-butyl ether (MTBE) (20 ml) and stirred for 2 h at 25 ± 5 ° C. The suspension was heated at 50 ° C for 3 h and then brought to 25 ± 5 ° C. N-Heptane (10 ml) was added and the reaction mixture was stirred for 72 h at 25 ± 5 ° C. The mass was then filtered and the obtained solid was dried at 50 ° C under vacuum for 12 h, to give Sofosbuvir form E (as confirmed by DPRX).

Example 12: Preparation of crystalline form E of Sofosbuvir

Sofosbuvir (Form 1, 1 g) was dissolved in t-butanol (3 ml). Then n-hexane (8 ml) was added, the mass was stirred for 24 h at 25 ± 5 ° C to provide a white solid. The mass was filtered and the obtained solid was dried at 50 ° C under vacuum for 12 h to provide Sofosbuvir E form (as confirmed by DPRX).

Example 13: Preparation of crystalline form E of Sofosbuvir

Sofosbuvir (Form 1, 1 g) was dissolved in methyl isobutyl ketone (MIBK) (4 ml). Cyclohexane (4 ml) was then added and the mass obtained was stirred for 24 h at 25 ± 5 ° C to provide a white solid. The mass was filtered and the obtained solid was dried at 50 ° C under vacuum for 12 h to provide Sofosbuvir E form (as confirmed by DPRX, Figure 1 and Figure 7).

X-ray powder diffraction (Examples 14 and 15)

The samples were measured on an X D8 Advance rat powder diffractometer (Bruker AXS, Karlsruhe, Germany) on a rotating PMMA sample holder (diameter: 25mm; depth: 1mm) in reflex mode (Bragg-Brentano geometry). Raw data was analyzed with the EVA program (Bruket AXS, Karlsruhe, Germany). Background subtraction and Ka2 pickling were not performed with the diffractograms shown in Figure 2 and Figure 8. Peak intensities (Figure 9) were determined after background subtraction.

Measurement conditions:

Cu Ka1 / a2 radiation

source 34 kV / 40 mA

Vantec-1 detector (electronic window: 3 °),

Kp Ni filter (diffracted beam)

diameter of measuring circle 435 mm

detector window slot 12 mm

anti-scattering slot (diffracted beam) 8 mm

divergence slot v6.00 (variable)

Soller slot (incident / diffracted beam) 2.5 °

interval 20 2 ° <20 <55 °

stage size 0.016

stage time 0.2 s

For Examples 14 and 15, Sofosbuvir was prepared according to the method described in B. S. Ross, P. G. Reddy, H.-R. Zhang, S. Rachakonda, M. J. Sofia, J. Org. Chem. 2011, 76, 8311. Form 1, described in US 2011/251152, was obtained by crystallization of the product from dichloromethane solution at room temperature. * Example 14: Preparation of crystalline form 7 of Sofosbuvir

Form 1 Sofosbuvir (200 mg) was stored as a powder in an open glass vial at 40 ° C / 75% relative humidity. After 4 weeks a solid amorphous had formed. This solid crystallized during continued storage at 40 ° C / 75% relative humidity after an additional 4 weeks. Form 7 was obtained as a colorless solid in quantitative yield.

X-ray powder diffractogram of Form 7 is illustrated in Figure 8.

* Example 15: Preparation of crystalline form 7 of Sofosbuvir

Sofosbuvir Form 1 (1.04 g) was heated in deionized water (40 ml) at 50 ° C for 2 hours with stirring. Partial dissolution occurred and the remaining solid turned into a sticky mass. Crystallization was not observed. Approximately 10 mg of Form 7 (obtained from the procedure of Example 14) was added and the mixture was cooled to room temperature. Form 7 crystallization occurred during overnight storage. A small amount of sticky mass remained on the stir bar, which was mechanically removed. Form 7 was isolated by filtration and dried in a desiccator evacuated over silica gel.

X-ray powder diffraction (Examples 16-20)

The samples were measured on a X D8 Advance rat powder diffractometer (Bruker AXS, Karlsruhe, Germany) on a rotating PMMA sample holder (diameter: 25mm; depth: 1mm) in reflex mode (Bragg-Brentano geometry). The measurement conditions are summarized in the table below. Raw data was analyzed with the EVA program (Bruket AXS, Karlsruhe, Germany). The background subtraction and etching of Ka2 were not performed with the diffractogram shown in Figure 10. The peak intensities were determined after background subtraction.

Reference Examples A-C for Examples 16-21:

*TO. Sofosbuvir starting material (Form 1)

Sofosbuvir Form 1 was prepared by crystallization of Sofosbuvir from dichloromethane according to the method described in B.S. Ross, P. G. Reddy, H.-R. Zhang, S. Rachakonda, M. J. Sofia, J. Org. Chem. 2011, 76, 8311 (page 8311). Sofosbuvir Form 1 is characterized by DPRX peaks at approximately 5.0, 7.3, 9.4, 18.1 ± 0.2 degrees 2-theta. Form 1 can be further characterized by additional DPRX peaks at about 8.8, 10.0, 11.4, 15.0, and 22.3 degrees 2-theta ± 0.2 degrees 2-theta.

* B. Preparation of Sofosbuvir Seed Crystals Form 7

(i) Small amounts of Sofosbuvir Form 7, which were used as seed crystals, were prepared as follows: Sofosbuvir Form 1 (200 mg) was stored as a powder in an open glass vial at 40 ° C / 75% relative humidity. After storage for 8 weeks at 40 ° C / 75% relative humidity, Form 7 was obtained as a colorless solid in quantitative yield.

(ii) Form 7 was prepared by heating Sofosbuvir Form 1 (1.04 g) in deionized water (40 ml) at 50 ° C for 2 hours with stirring. Partial dissolution occurred and the remaining solid turned into a sticky mass. Crystallization was not observed. Approximately 10 mg of Form 7 [obtained from (i)] was added and the mixture was cooled to room temperature.

Form 7 crystallization occurred during overnight storage. A small amount of sticky mass remained on the stir bar, which was mechanically removed. Crystalline Form 7 was isolated by filtration and dried in a desiccator evacuated over silica gel.

* C. Preparation of water-saturated MTBE (methyl tere-butyl ether)

Water-saturated MTBE was prepared by vigorous shaking (500 rpm) of MTBE with deionized water in a volume ratio of 10: 1 for 30 minutes at room temperature and subsequent separation phase to remove the aqueous layer. The organic phase was prepared fresh before use. Alternatively, the organic phase can be stored in a sealed container at room temperature.

Example 16 (Preparation of Sofosbuvir Form 7)

Sofosbuvir (50 g) was dissolved almost completely in water-saturated MTBE (800 ml) at room temperature. The solution was filtered through a pleated filter. Sofosbuvir Form 7 seed crystals (150 mg) were added and the solution was stored for two days in a closed flask at room temperature without stirring. The crystals formed (Form 7) were isolated by filtration, washed with a small amount of MTBE and dried at room temperature under vacuum (2 mbar) overnight. The yield was 23.9 g. The mother liquor was concentrated to a volume of 350 ml on a rotary evaporator and then stored for two days in a closed flask at room temperature without stirring. During this time, the initially separated oil crystallized. The solid formed (Form 7) was then isolated by filtration, washed with a small amount of MTBE and dried at room temperature under vacuum (2 mbar) overnight. The weight of the solid (Form 7) was 15.7 g. Therefore, the combined yield of Form 7 was 39.7 g (79%).

* Example 17: Preparation of Sofosbuvir Form 6, small scale

Sofosbuvir (2 g) was dissolved almost completely in water-saturated MTBE (30 ml) at room temperature. The The solution was filtered through a pleated filter and stirred magnetically (400 rpm) for 12 h in a closed flask at room temperature. The crystals formed (consisting of Form 6) were isolated by filtration, washed with a small amount of MTBE and dried at room temperature under vacuum (2 mbar) overnight. The yield was 1.2 g (60%).

* Example 18: Preparation of Sofosbuvir Form 6)

Sofosbuvir (40 g) was dissolved almost completely in water-saturated MTBE (600 ml) at room temperature. The solution was filtered through a pleated filter. Sofosbuvir Form 6 seed crystals (100 mg, obtained from Example 17) were added and the solution was stirred mechanically (300 rpm) for 12 h in a closed flask at room temperature. The crystals formed (Form 6) were isolated by filtration, washed with a small amount of MTBE and dried at room temperature under vacuum (2 mbar) overnight. The yield was 29.6 g (74%).

* Example 19: Dissolution of Form 6 and Form 7 of Sofosbuvir

Sofosbuvir Form 6 was used according to Example 18 above and Sofosbuvir Form 7 according to Example 16. Dissolution of Sofosbuvir Form 7 and Form 6 was measured (in triplicate) in 50 mM phosphate buffer at 37 ° C (pH 6.8, 900 ml) using apparatus II USP, paddle method. Every 5 minutes, a probe was taken and measured by UV / VIS at 260 nm and a path length of 0.1 cm; UV / VIS was measured on an Agilent 8453 with Chemical Station Dissolution Software. 400 g of Sofosbuvir Form 7 and Form 6 were used for the assay and the percentages are given relative to 400 g of the PFA. The results are graphically represented in Figure 13.

* Example 20: Solubility and Particle Size of Sofosbuvir Form 6 and Form 7

Sofosbuvir form 6 was used according to Example 18 above and Sofosbuvir form 7 according to Example 16.

Particle size distribution was determined using a wet measurement using a Malvern Instruments 2000S Mastersizer 2000 and Malvern Instruments wet sample of the dispersion unit and silicone oil as dispersant. The following settings are used:

The results are shown in the following table:

Tabl: T m ñ ^ rí l m F rm 7 F rma 6

The solubility was determined as follows:

100 mg of test substance was weighed into a glass vial, followed by the addition of 4 ml of solvent (water, 0.01 M HC1.20 mM sodium acetate / acetic acid [pH 4.5], 50 mM KH2PO4 [pH 6.8]). A stir bar was added, the vial was set in a block heater at 37 ° C, and the suspension was stirred at 250 rpm. After 15 min and 1 h, the samples were removed through a 0.2 pm PTFE filter. 100 µl of the clear filtrate was diluted with 900 µl of acetonitrile / water mixture (1: 1) and 1 µl of it was analyzed by UHPLC / UV:

Instrument: Agilent 1290 Infinity

Wavelength: 260 nm

Column: Phenomenex Kinetex XB - C18, 1.7 pm, 50 x 3 mm

40.0 ° C temperature

column:

Flow [ml / min]: 0.4

Volume of

injection: 1 pl

water 0.2% formic acid 0.1% HFBA (acid

Solvent A: heptafluorobutyric)

Solvent B: acetonitrile

Time gradient [min] Solvent B [%]

(continuation)

0.00 30.0

7.00 70.0

7.10 30.0

10.00 30.0

The results are shown in the following table:

T l: l ili m ml F rm F rm 7 f vir 1 min 1 h 7 °

The solubility of Form 7 was measured in physiologically relevant solvents at 37 ° C after 15 min and after 1 hour it is one third higher than the solubility of Form 6. This result is particularly surprising in view of the fact that the size Particle size of the examined batch of Form 7 is distinctly larger than that of Form 6. Thus, Form 7 provides a significant advantage with respect to its solubility behavior.

* Example 21: Sofosbuvir Form 7 Single Crystal Data

Sofosbuvir Form 7 crystals were obtained by the procedure of Example 16.

A small prism was selected as a suitable identical crystal, since a sufficient volume of crystal could not be found. Data was collected on a Gemini Atlas CCD (Cu Ka) diffractometer using u> scans. Data collection runs were optimized for absolute configuration analysis. The positional and anisotropic thermal parameters of all non-hydrogen atoms were refined. The carbon-bound H atoms were arranged geometrically. The positions of the H atoms were refined with gentle restrictions on bond lengths and angles to regularize their geometry (in the C-H range 0.93-0.98). The positions of hydrogen atoms attached to nitrogen and oxygen were initially refined with restricted bond lengths and then with conduction limitations. Uiso (H) was established in the range of 1.2-1.5 times Uec of the parent atom. The contributions of twin component 1, twin component 2, inverted component 1, and inverted component 2 were refined to 0.60 (4), 0.42 (3), 0.00 (4), and 0.03 (3), by therefore, the absolute configuration mapping is correct. No accessible residual solvent voids were disclosed by the Platon gap calculation routine. Software used:

Data collection: CrysAlisPro CCD (Oxford Diffraction Ltd, 68 Milton Park, Abingdon, Oxfordshire OX144RX, England, Version 171.31.7, 2002);

cell refinement: CrysAlisPro RED;

data reduction: CrysAlisPro RED;

program used to dissolve the structure: Superflip [Palatinus L. & Chapuis G. (2007). J. Appl. Cryst. 40, 786,790];

program used to refine the structure and absolute configuration analysis: Crystals [Betteridge, PW, Carruthers, JR, Cooper, RI, Prout, K. & Watkin, DJ (2003). J. Appl. Cryst. 36, 1487]; Molecular chart: Mercury; Vacuum calculation was performed using PLaton [Spek, AL (2003). PLATON, A multipurpose crystallographic tool, Utrecht University, Utrecht, The Netherlands]

continuation

Example 22- Pharmaceutical compositions (for example, tablet) of sofosbuvir

Preparation method:

Batches were prepared by a dry granulation method. The intragranular ingredients from Part I were mixed for 10 minutes, Magnesium stearate from Part II was added to the mixture and mixed for an additional 5 minutes. The mixture obtained was granulated by means of the roller compactor and ground through a 0.8 mm screen and mixed with Part II of extra-granular excipients for 10 minutes. Magnesium Stearate Part IV was added to the mixture and mixed for an additional 5 minutes. The final mixture was compressed into tablets.

Physical results

The Hausner and Carr relationship is often used as an indication of the flowability of a powder. The Hausner factor is the ratio of mass volume to compacted volume, calculated by the volume density / compressed density formula. Volume and compressed density are measured according to the general pharmacopoeia method. The Hausner ratio of <1.25 indicates a free flowing powder. A Carr index of> 23 indicates poor flow.

Claims (13)

1. Crystalline form of Sofosbuvir designated Form E and characterized by data selected from one or more of the following: an X-ray powder diffraction pattern having peaks at: 12.4, 16.2, 17.2, 25, 0 and 25.3 degrees two theta ± 0.1 degrees two theta; an X-ray powder diffraction pattern as illustrated in Figure 1 or Figure 7; an X-ray powder diffraction pattern having peaks at: 12.4, 16.2, 17.2, 25.0 and 25.3 degrees two theta ± 0.1 degrees two theta and no peaks at: 10 , 9 and 14.2 degrees two theta ± 0.2 degrees two theta, and combinations of these data. 2. Crystalline Form E of Sofosbuvir according to Claim 1, wherein the crystalline form is further characterized by data selected from one or more of the following: X-ray powder diffraction pattern having peaks at 12.4, 16 , 2, 17.2, 25.0 and 25.3 degrees two theta 0.1 degrees two theta and also has one, two, three or four peaks selected at: 8.1, 19.4, 22.0 and 23.3 degrees two theta 0.1 degrees two theta; an X-ray powder diffraction pattern having peaks at: 12.4, 16.2, 17.2, 25.0 and 25.3 degrees two theta 0.1 degrees two theta, no peaks at: 10, 9 and 14.2 degrees two theta 0.2 degrees two theta and also having one, two, three or four peaks selected from: 8.1, 19.4, 22.0 and 23.3 degrees two theta 0.1 degrees two theta. 3. Crystalline Form E of Sofosbuvir according to any one of Claims 1-2 which is characterized by the following X-ray powder diffraction peaks:

4. Crystalline Form E of Sofosbuvir according to any one of Claims 1-3, wherein Form E of Sofosbuvir is isolated. 5. Crystalline Form E of Sofosbuvir according to any one of Claims 1-4, which is anhydride. 6. Crystalline Sofosbuvir Form E according to any one of Claims 1-5, which is substantially free from any other form of sofosbuvir, preferably which is substantially free from Form 6 of sofosbuvir, as measured by XRPD, preferably in where Form 6 is characterized by having peaks at 6.08 and 10.85 degrees two theta 0.2 degrees two theta. 7. Crystalline Sofosbuvir Form E according to Claim 6, containing 20% or less, 10% or less, 5% or less, 2% or less, 1% or less of Form 6 sofosbuvir, preferably as measured by XRPD. 8. A pharmaceutical composition comprising crystalline Form E of Sofosbuvir according to any one of Claims 1-7. 9. A pharmaceutical composition according to Claim 8, wherein the pharmaceutical composition is a solid composition and the Sofosbuvir retains its form in the solid state. 10. A process for the preparation of a pharmaceutical composition according to any one of Claims 8-9, which comprises combining the crystalline form of Form E of Sofosbuvir according to any one of Claims 1-7 with at least one pharmaceutically acceptable excipient. 11. A crystalline form of Sofosbuvir according to any one of Claims 1-7 for use as medicine. 12. A crystalline form of Sofosbuvir for use according to Claim 11, wherein the medicament is for the treatment of Hepatitis C. 13. Use of a crystalline form of Sofosbuvir according to any one of Claims 1-7, in the manufacture of a pharmaceutical composition.