ES2717254T3 - Solid state forms of sofosbuvir - Google Patents

Description

DESCRIPTION

Solid state forms of sofosbuvir

Field of the invention

The present invention encompasses a solid state form of Sofosbuvir, processes for its production 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-methyl ethyl 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, having the following formula,

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

Sofosbuvir is marketed under the registered trade name of SOVALDI ®. SOVALDI ® is available as immediate release tablets. Each tablet contains 400 mg of sofosbuvir. The loading of active principle 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 ingredients: polyethylene glycol, polyvinyl alcohol, talc, titanium dioxide and yellow iron oxide. The documents 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. The solid state forms of Sofosbuvir are described in WO 2010/135569, US 2011/251152, WO 2011/123645 and CN 104130302. Documents 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 (eg, measured by thermogravimetric analysis - "TGA", or differential scanning calorimetry - "DSC"), pattern X-ray diffraction, infrared absorption fingerprint and solid state NMR (13C-) spectrum. One or more of these techniques can be used to distinguish different polymorphic forms of a compound.

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

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

Various salts and solid state forms (including solvated forms) of a pharmaceutical active ingredient may have different properties. Such variations in the properties of different salts and solid state forms and solvates can provide a basis for improving the formulation, for example, by facilitating better processing or manipulation of the characteristics, by changing the dissolution profile in a favorable direction or by improving the stability (polymorph 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 Various salts and solid state forms and solvates of a pharmaceutical active ingredient can also cause a variety of polymorphs and crystalline forms, which can, 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 the conversion into 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. Enlarge the repertoire of materials that a formula scientist has available for the optimization of formulations, for example, 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 useful life (chemical / physical stability). For example, it has now been found that Sofosbuvir Form 6 has a high tendency to become electrostatically charged. The electrostatically charged active pharmaceutical ingredients can exhibit poor fluidity and / or tendency to stick and, in this way, can ultimately result in complications during the operations of the manufacturing process of a pharmaceutical composition based on such an electrostatically charged pharmaceutical active ingredient. . In addition, poor content uniformity can be observed in the final dosage form when a dry process such as, for example, dry compression is used 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 Sofosbuvir forms, such as those described above, are, in general, impractical for the large-scale preparation medium. On the other hand, the resulting products may 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, as well as new processes for producing crystalline forms of Sofosbuvir. The processes described in this document allow the consistent production of these Sofosbuvir forms that can be used on a large scale. The present processes also seek to avoid the need to use a number of different organic solvents, which can cause unpredictable and unpredictable polymorphic transformation, and which can lead to the production of mixtures of different crystalline forms and, in addition, can introduce unwanted impurities in the the product.

Summary of the invention

The present invention provides a solid state form of Sofosbuvir, processes for its preparation and pharmaceutical compositions thereof. In particular, the present invention provides the crystalline E-form of sofosbuvir, a process for preparing the crystalline E-form of sofosbuvir, wherein the process comprises:

(a) combining sofosbuvir with a solvent system comprising an organic solvent and optionally water, wherein the organic solvent is selected from the group consisting of aliphatic ethers - preferably C4-C8 ethers, cyclic ethers, ketones - preferably C3-C8 ketones , alcohols - preferably C4-C8 alcohols and, preferably wherein the organic solvent is selected from the group consisting of methyl t-butyl ether, t-butanol, acetone, methyl isobutyl ketone and, preferably, methyl isobutyl ketone;

(b) optionally seeding with crystalline E-form;

(c) adding an anti-solvent;

(d) optionally isolating the crystalline E form; Y

(e) optionally deagglomerating or triturating or micronising to obtain the crystalline E form of sofosbuvir having a smaller particle size;

wherein Form E is 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 absence of 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.

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

The present invention also provides Form E of sofosbuvir and compositions thereof for the treatment of Hepatitis C.

BRIEF DESCRIPTION OF THE DRAWINGS

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

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

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

Figure 4 shows the isothermal chart of DVS of Sofosbuvir form 1

Figure 5 shows the Isothermal Chart of DVS of Sofosbuvir Form 6

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

Example 7 - DPRX of Sofosbuvir Form E at 2-30 degrees of 2-theta interval

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

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

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

Figure 11 shows the diffraction peaks of X-ray powder for Sofosbuvir Form 7 (Example 1) and the relative intensities measured using Ka1 / Ka2 copper radiation with a weighted median wavelength of 1.54187 A. In this table, the 2-theta values are reported with an accuracy of 0.2 degrees 2-theta. Figure 12 shows the X-ray powder diffraction peaks of Sofosbuvir Form 1 and the relative intensities. In this table, the 2-theta values are reported with an accuracy of 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 II apparatus, 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 the control of the conditions in which Sofosbuvir is obtained in solid form.

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

As used herein, "substantially free" means that the solid state forms of the present invention contain 20% (w / w) or less, 10% or less, 5% or less, 2% or less than 1% or less of any other form of the subject compound or of a specified polyphosphor 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 of a specified polymorph of Sofosbuvir. In other embodiments, the solid state form of Sofosbuvir of the present invention contains from 1% to 20% (w / w), from 5% to 20% (w / w) or from 5% to 10% (w / w) ) in any solid state form of a specified polymorph of Sofosbuvir.

Depending on what comparison with other forms in the solid state is performed, the Sofosbuvir solid / crystalline form 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 stability 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, fluidity and advantageous processing and characteristics of handling such as compressibility and density in mass.

A solid state form, such as a crystal shape or amorphous form, may be referred to herein as being characterized by graphic data "as illustrated in" or "as substantially illustrated in" a Figure. Such data include, for example, powder X-ray diffractograms. As is well known in the art, the graphic data potentially provides additional technical information to further define the respective solid state form (a so-called "fingerprint") that can not necessarily be described by reference to numeric 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 be easily able to compare the graphic data in the Figures in the present document with graphic data generated for a solid / crystalline form. unknown and confirm whether the two sets of graphical data characterize the same form in solid / crystalline state or two forms in solid / crystalline state. A solid / crystalline 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 in solid / crystalline state of Sofosbuvir characterized with graphic data having such small variations, as are well known by the experts, in comparison with the Figure.

As used herein and unless otherwise indicated, the term "anhydride" with respect to solid state / crystalline forms of Sofosbuvir, refers to a solid / crystalline form of Sofosbuvir that does not include water (or other solvents) in a defined, stoichiometric amount within the crystal. On the other hand, an "anhydride" form would typically not contain 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 Sofosbuvir solid state forms 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 stated, measurements of DPRX are taken using a Ka radiation of copper with a wavelength of 1.5418 A. Unless otherwise indicated, the values of 2 -theta of DPRX are released with an error of 0.2 degrees 2-theta.

The individual particles of a sample or aliquot of the solid particles comprising the crystalline E-form of Sofosbuvir of the present invention are not uniform in size. In contrast, a sample or aliquot of a solid particle comprising Sofosbuvir crystalline E-form of the present invention is comprised of different sizes that can be sorted by size or distributed in a grouping of discrete 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 samples of particles comprising the crystalline E form of Sofosbuvir based on individual points of a cumulative particle size distribution curve. The measurements are represented as d (0, X) = Y (where X and Y are Arabic numerals), each "d" describes an individual point of a cumulative DPT curve. The number "X" represents the percentage (number, volume or weight) of particles in the population that has a nominal size of up to and that includes "Y". Therefore, d (0.9) = 250 p is characteristic of a PTD in which 90% (number, volume, or weight) of the particles in a population has a nominal size of approximately 250 po menso (at least some particles that have a nominal dimension of 250 p), and so on. When DPT is determined by the well-known laser diffraction method that is described herein, the measurement d (0, X) illustrates a volume average.

The person skilled in the art knows that the results of the determination of DPT by one technique can be correlated with results of another technique on an empirical basis by means of routine experimentation.

As used herein, unless otherwise indicated, "medium particle size" refers to the D ⁵⁰ value of the particle size distribution. As used herein, the particle size distribution is determined by laser diffractometry. More specifically, unless otherwise indicated, the particle size was determined using a Mastersizer 2000 from Malvern Instruments - the determination of particle size can be carried out as a wet or dry measurement depending on the sample.

The median particle size (D ⁵⁰ ), which is also denoted D50 value of the integral volume distribution, is defined in the context of the present invention as the particle diameter, in which 50 volume percent of particles have a diameter less than the diameter that corresponds to the D ⁵⁰ value. Also, 50 percent of the volume of the particles has a diameter greater than the D ⁵⁰ value. Analogously, the D ⁹⁰ value of the integral volume distribution is defined as the particle diameter, in which 90 percent by volume of particles have a diameter smaller than the diameter, which corresponds to the D ⁹⁰ value. Correspondingly, the D ¹⁰ value of the integral volume distribution is defined as the particle diameter, wherein the 10 percent by volume of particles have a diameter smaller than the diameter, which corresponds to the D ¹⁰ value.

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

An object, for example, a reaction mixture can be characterized herein as being at, or allowed to return to "room temperature" or "ambient temperature", unless abbreviated as "TA". 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 smoke bell, in which the object is located. In general, 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 used in a chemical process, for example, a reaction or a crystallization, can 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 expression would be understood to refer to milliliters of the solvent per gram of the material being suspended, so that the suspension of 5 grams of a material in 10 volumes of a solvent means that the solvent is used in an amount of 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 can be referred to in the present document as being carried out "overnight." This refers to the time interval, for example, for the process or stage, which occurs in the time during the night, when a process or stage can not be observed actively. This time interval is from about 8 to about 20 hours, or about 10-18 hours, normally about ¹⁶ hours.

As used herein, the term "reduced pressure" refers to a pressure that is lower 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 "under vacuum" refers to a pressure of from about 0.2 mbar to about 10 mbar, from about 0.2 to about 5 mbar, of about 0.5 to about 4 mbar, from about 1 mbar to about 3 mbar and preferably about 2 mbar.

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

As used herein, unless otherwise indicated, the term "compressibility" is expressed as maximum density at a 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. The crystalline form of Sofosbuvir can be characterized by DpRX peaks of 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 Therefore, the Sofosbuvir crystalline 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 form of Sofosbuvir prepared by a process according to any embodiment of the present invention may be substantially free of any other form of Sofosbuvir or the specified polymorphic forms of Sofosbuvir, respectively.

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 absence of 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 between: 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, absence of peaks at: 10 , 9 and 14.2 degrees two theta ± 0.2 degrees two theta and that also has 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 having peaks in 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 one absence of peaks in 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 following table.

:

In another embodiment, the crystalline E form 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 X-ray powder diffraction pattern having peaks in: 8.1, 12.1 , 12.4, 13.5, 16.2 and 17.2 degrees two theta ± 0.1 degrees two theta.

In another embodiment, the E-shape 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 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 the E-form of sofosbuvir wherein Form E is characterized by an X-ray diffraction pattern having peaks in 8.1, 12.1, 12, 4, 13.5, 16.2 and 17.2 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 another embodiment, the crystalline E form 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 two degrees 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 X-ray powder diffraction pattern having peaks in: 8.1, 10.4 , 13.5, 18.0 and 18.7 degrees two theta ± 0.1 degrees two theta.

In another embodiment, the E-shape 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 the E form of sofosbuvir wherein Form E is characterized by an X-ray diffraction pattern having peaks in 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 one embodiment Form E is anhydride.

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

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

The compositions comprising the crystalline E-form of the present invention are preferably polymorphically pure, ie, they are 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 sofosbuvir form 6 measured by DRXP. Accordingly, the content of Sofosbuvir crystalline form 6 in Sofosbuvir crystalline E-form will be measured by detecting and quantifying the characteristic peaks described in form 6. The characteristic peaks of Sofosbuvir crystalline 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, i.e. it is less hygroscopic in comparison with 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

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

The improved hygroscopic properties described hereinbefore can be expressed by the dynamic water absorption controlled by the DVS instrument. The profile of the dynamic curve obtained can provide insight into the tendency of the substance to deliquescence. This is expressed by a sudden change in the slope of the curve (point of inflection) 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 sticking and forming 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). A 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 Form E of low hygroscopicity does not show this inflection point. In addition, this form E shows no 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 is formed between the value of increase of weight of absorption and the value of decrease of weight of desorption at a given relative humidity. A comparison of the water content values as measured by DVS is shown in Table 2 above.

The Sofosbuvir crystalline form of the invention can be characterized by an isothermal water vapor sorption which shows 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 Sofosbuvir crystalline form of the invention can be characterized by an isothermal water vapor sorption which shows a water absorption of not more than 0.10% by weight, a relative humidity (RH) of up to 70% and / or an absorption of water 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 present invention can have a low hygroscopicity characterized by an isothermal water vapor sorption which shows a water absorption of not more than 0.10% by weight, a relative humidity (RH) of up to 70% and / or 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%.

In another aspect, the invention relates to a process for preparing the crystalline E-form of sofosbuvir comprising (a) combining sofosbuvir with a solvent system comprising an organic solvent and optionally water (b) optionally plating with Form E crystals ( c) adding an antisolvent (d) optionally isolating the crystalline E form and (e) optionally grinding or micronising to obtain the crystalline E form of sofosbuvir having a lower particle size. The process may also comprise filtering and drying steps.

The organic solvents in step (a) is 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 an anti-solvent 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 anti-solvents can be selected from a group consisting of di-isopropyl ether, n-heptane, cyclohexane. Most preferably, the antisolvent is cyclohexane.

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

Preferably, the reaction mixture of step (a) includes from about 10% to about 50% Sofosbuvir, by volume. Preferably, the process according to the present invention 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 of the present invention is carried out with stirring

Generally, the amount of crystalline E form of Sofosbuvir used for seeding in step (b) is about 0.1% to 0.5% by weight of 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 E form of Sofosbuvir. Preferably, the crystalline E form 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 crushed or micronized to obtain sofosbuvir that has a smaller particle size in a process that adapts to the desired particle size. For example, micronization can be carried out using a 100 mm plate air jet micronizer.

In another aspect, the invention provides a process for preparing the crystalline E form of Sofosbuvir comprising: a) combining Sofosbuvir with methyl isobutyl ketone and water ab) seeding with E-shaped crystals, c) adding cyclohexane d) optionally isolating form E and (e) optionally grinding or micronizing to obtain the crystalline E form of Sofosbuvir having a desired particle size. The process may also comprise filtering and drying steps.

In one embodiment, the starting material used in the above-described process is crystalline form 1 of Sofosbuvir.

In another embodiment, 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.

In yet another embodiment, step a) is carried out at a temperature ranging from 0 to 20 ° C, preferably, step a) is carried out at 9 ± 3 ° C.

In a preferred embodiment, the invention provides a process for preparing the crystalline E form of Sofosbuvir comprising: a) combining Sofosbuvir with methyl isobutyl ketone and water wherein the amount of water is 1% by volume and the temperature is 9 ± 3 ° C b) seeding with E-shaped crystals, c) adding cyclohexane d) isolating Form E and (e) optionally micronizing. The process may also comprise filtering and drying steps.

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

In the most preferred embodiment, crystalline form 1 of sofosbuvir is combined with MINK 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 loaded, stirred at at 9 ± 3 ° C, seeded with crystals of form E and stirred as such for 4 ± 0.5 h. Next, cyclohexane is 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 obtained solid was dried under vacuum for 12 h at Tj 40-50 ° C to obtain Sofosbuvir Form E.

In another aspect, the invention relates to the crystalline E-form produced by the process described above.

In another aspect, the invention encompasses micronized or small particle size form of sofosbuvir having a desired DTP which 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.

The sofosbuvir of small particle size can be directly obtainable by different processes, for example, it can be obtained by chemical synthesis or by micronization techniques, optionally, by the use of deagglomeration techniques.

Micronization is a technique known to those skilled 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 micronised or small particle size sofosbuvir comprising crystalline E-form of the present invention shows good fluidity and compressibility and reduced hygroscopicity, therefore, it can be used advantageously for the preparation of pharmaceutical compositions and formulations.

The physical attributes of PFA are especially important for the reproducibility of preparations of dosage forms 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 E form of micronized Sofosbuvir or small particle size of the present invention is suitable for use in the preparation of formulations containing a high sofosbuvir load, for example, containing more than 35% sofosbuvir. In addition, the micronized sofosbuvir crystalline form E or sofosbuvir small particle size of the present invention 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 principle, for example, ledipasvir.

In one embodiment, the invention encompasses micronized or small-particle Sofosbuvir comprising crystalline E-form of Sofosbuvir having preferred flow properties, ie, showing a flow rate of only 100 s / 100 g, more preferably no more than 80 s / 100 g, more preferably no more than 65 s / 100 g, as measured by an orifice flow in the automatic powder testing system of Pharma-Test 41-03300.

In another embodiment, the invention encompasses Sofosbuvir micronized or small particle size of Sofosbuvir crystalline E-form having advantageous compressibility properties, ie, showing a constant density not lower 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.

In another embodiment, the invention encompasses Micronized Sofosbuvir or small particle size of Sofosbuvir crystalline E-form characterized by a water sorption of no more than 0.5%, more preferably no more than 0.3% as measured by DVS and water content after balancing at 90% RH.

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

In yet another preferred embodiment, the invention encompasses micronized crystalline E form or small particle size characterized by DTP of d (0.1) 0.4-0.6 microns, d (0.5) 1.0-2, 0 micrometers and d (0.9) 3.2-5.0 microns; a flow rate of no more than 100 s / 100 g, more preferably no more than 80 s / 100 g, more preferably no more than 65 s / 100 g; a constant density of 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.

In the particularly preferred embodiment, the invention encompasses micronized crystalline E form or small particle size characterized by DTP of d (0.1) 0.4-0.6 microns, d (0.5) 1.0-2, 0 micrometers and d (0.9) 3.2-5.0 microns; a flow rate of not more than 65 s / 100 g; a constant density not lower 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 micronized or small particle size form described above of Sofosbuvir can be used to prepare pharmaceutical compositions and formulations.

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

The above crystalline E form of Sofosbuvir of the present invention can also be used as medicaments.

The present invention additionally encompasses 1) the use of the crystalline form of Sofosbuvir mentioned above in the manufacture of a pharmaceutical composition and 2) a method of treating a subject suffering from Hepatitis C or otherwise in need of treatment, which comprises administering an effective amount of a pharmaceutical composition comprising the above crystalline Sofosbuvir form described herein.

The 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.

The pharmaceutical compositions in the form of a tablet according to the present invention are preferably for use in the treatment of 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 granulated dry or wet granulated prior to tableting or tablet mixtures can be compressed directly. Granulation is preferred for the preparation of sofosbuvir tablets and dry granulation is most preferred. The 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, first a tablet is formed, then the tablet is ground to obtain particles that are easier to handle than the original mixture. The comminuted 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 composition according to the present invention comprises sofosbuvir of small particle size, preferably the crystalline sofosbuvir of small particle size is the E-form of small crystalline particle size of sofosbuvir. In another embodiment, the pharmaceutical composition comprises sofosbuvir of small particle size and without glidants.

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

The composition comprising sofosbuvir of small particle size 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 no more than 1.5% magnesium stearate, more preferably no more than 1%, most preferably no more than 0.6% magnesium stearate based on the weight of the total composition. For pharmaceutical compositions comprising sofosbuvir of small particle size and without glidants, the particle size distribution is measured by laser diffraction and preferably refers to a volume distribution. Preferably, the particle size distribution is measured using the Malvern Mastersizer 2000. The sofosbuvir of small particle size preferably has a d (0.5) of less than 20 μm, preferably less than 10 μm, more preferably less than 5 pm, and most preferably between 1-2 pm. In one embodiment, the sofosbuvir of small particle size 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. Reference is also made to Example 22 for the measurement of sofosbuvir particle size.

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

Methods

Diffraction of X-ray powders (Examples 1-13)

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

Measurement parameters:

Scanning interval: 2-40 degrees 2-theta;

Scan mode: continuous;

Stage size: 0.05 degrees;

Time per stage: 0.5 s;

Rotation of the sample: 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 peak (Si) 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 length: 720 min

Fluency

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

Method of DTP measurements 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 Mastersizer 2000 from Malvern Instruments and wet sample from Malvern 2000S of the dispersion unit and silicone oil as the dispersant. Unless indicated otherwise, the following configurations are preferably used:

Method of measuring DTP of the form E-Measurement of dry cell

Mastersizer 2000

Analysis model: General purpose

IR of powder: 1,573

Absorption: 0.5

Dispersant name: Dry dispersion (air)

IR of the dispersant: 1.0

Sensitivity: Normal (fine powder)

Filtering obscuration: On (0.5-3%)

Measuring time: 5 seconds (5000 snapshots)

Background measurement time: 5 seconds (5000 snapshots)

Aliquot repetitions: 1

Repetition measurement: 1

Pressure: 2.5 Bar Feed rate of the flow cell: 30%

Funnel recess: 5 mm

Feeder with small sieve filled with balls (2/3 by volume)

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

A background measurement should be made before each measurement. Add approximately 0.5 g of sofosbuvir powder with a spatula directly to the feeder. Start the work of the feeder. The 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 E form of Sofosbuvir

Sofosbuvir (1 g, Form 1) was dissolved in acetone (3 ml) with stirring at 25 ± 5 ° C. Next, 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 E-form of Sofosbuvir (as confirmed by DPRX).

Example 2: Preparation of crystalline E form of Sofosbuvir

Methyl iso-butyl ketone (MIBK) (3 ml) in 100 ml was loaded into a round-bottomed 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 became slowly transparent after 15 min at 0 to 5 ° C. At this point, Sofosbuvir Form E cultures (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 h at 0 to 5 ° C. Cyclohexane (10 ml, pre-cooled to 15 ° C) was added and stirred for 2 to 5 min. The precipitate was filtered and washed with cyclohexane (10 ml, pre-cooled at 15 ° C). The solid was dried at 33 ° C under vacuum for 12 h, to provide E-form of Sofosbuvir (as confirmed by DPRX).

Example 3: Preparation of crystalline E form of Sofosbuvir

Charge (MIBK) (60 ml) in 250 ml in a round-bottomed glass flask equipped with a mechanical stirrer. This was cooled to 10 to 15 ° C with stirring and Sofosbuvir (20 g, Form 1) was added at 10 to 15 ° C. This mass became slowly transparent after 20 min at 10 to 15 ° C. At this point, Sofosbuvir Form E cultures (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 h at 10 to 15 ° C. Cyclohexane (60 ml, pre-cooled to 15 ° C) was slowly added and stirred for 2 to 5 min. The precipitate was filtered and washed with cyclohexane (60 ml, pre-cooled at 15 ° C). The obtained solid was dried at 33 ° C under vacuum for 12 h, to provide E-form of Sofosbuvir (as confirmed by DPRX).

Example 4: Preparation of crystalline E form of Sofosbuvir

Charge (MIBK) (3 ml) in 100 ml in a round-bottomed glass flask equipped with a mechanical stirrer. This was cooled to 10 to 15 ° C with stirring and Sofosbuvir (1 g, Form 1) was added at 10 to 15 ° C. This mass became slowly transparent after 15 min at 10 to 15 ° C. At this point, Sofosbuvir Form E cultures (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 obtained mass was stirred for 1 h at 10 to 15 ° C. Cyclohexane (10 ml, pre-cooled 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 E-form of Sofosbuvir (as confirmed by DPRX).

Example 5: Single-crystal X-ray crystallography of Form E

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

Parameters of Sofosbuvir crystal lattice, Form E:

length_celda_a 14,4723 (2) A

length_cell_b 17,0212 (2) A

length_celda_c 5,22862 (6) A

angle_alpha_alpha 90.0000 °

angle_cell_beta 100,297 (1) °

90.0000 ° angle_celda_gamma

cell-volume 1267.3 A3

group_spacio_symmetry_name: P21

monoclinic_cell_symmetry configuration

temperature_measure_cell 293 K

Example 6: Preparation of crystalline E form of Sofosbuvir

Form 1 of Sofosbuvir (100 g) was loaded together with MIBK (500 ml) in 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. Form E crystals of Sofosbuvir (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 white solid obtained was washed with cyclohecano (2 X 200 ml) and sealed at 45 ° C under vacuum for 12 h to provide an E-form of sofosbuvir (as confirmed by DPRX).

Example 7: Preparation of crystalline E form 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 becomes clear. The solution was filtered to remove any foreign particles. It was washed with MIBK (0.5 l). The filtered solution was loaded, 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 crystalline E form of micronized Sofosbuvir

Micronized sofosbuvir crystalline E form was prepared using a 100 mm plate air jet micronizer, feeding speed 0.5 Kg / h, feed pressure 6.5 bar +/- 0.5 bar, crushing pressure 6 bar +/- 0.5 bar.

Example 9: Fluidity of crystalline E form of micronized sofosbuvir

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

Example 10: Compressibility of crystalline E form of micronized sofosbuvir

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

200 mg of the crystalline E form of micronized Sofosbuvir obtained according to example 8 is compressed in an aceto mold (with the displacement rate of 0.03 mm / sec). A cyclic process (similar to screw) was carried out: compress, then refract, relax for 15 s and then repetitive compressive steps (in 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 E form of Sofosbuvir

Sofosbuvir (Form 1, 1 g) was suspended in methyl fer-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 solid obtained was dried at 50 ° C under vacuum for 12 h, to provide E-form of Sofosbuvir (as confirmed by DPRX).

Example 12: Preparation of crystalline E form of Sofosbuvir

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

Example 13: Preparation of crystalline E form of Sofosbuvir

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

Diffraction of X-ray powders (Examples 14 and 15)

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

Measuring conditions:

Cu Ka1 / a2 radiation

34 kV / 40 mA source

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

Kp Ni filter (diffracted beam)

Measuring circle diameter 435 mm

12 mm detector window slot

anti-dispersion 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 0.2s

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 the 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 amorgo had formed. This solid crystallized during continued storage at 40 ° C / 75% relative humidity after an additional period of 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

Form 1 Sofosbuvir (1.04 g) was heated in deionized water (40 ml) at 50 ° C for 2 hours with shaking. Partial dissolution occurred and the remaining solid became a sticky mass. No crystallization was observed. Approximately 10 mg of Form 7 (obtained from the procedure of Example 14) was added and the mixture was cooled to room temperature. The crystallization of Form 7 occurred during its storage overnight.

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

Diffraction of X-ray powders (Examples 16-20)

The samples were measured in an X D8 Advance powder diffractometer (Bruker AXS, Karlsruhe, Germany) on a rotating PMMA sample holder (diameter: 25 mm, depth: 1 mm) in reflex mode (Bragg-Brentano geometry). The conditions of the measurements are summarized in the table below. The raw data was analyzed with the EVA program (Bruket AXS, Karlsruhe, Germany). The background subtraction and pickling of Ka2 were not performed with the diffractogram shown in Figure 10. 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 approximately 8.8, 10.0, 11.4, 15.0 and 22.3 degrees 2-theta ± 0.2 degrees 2-theta.

* B. Form 7 preparation of Sofosbuvir seed crystals

(i) Small amounts of Sofosbuvir Form 7, which were used as seed crystals, were prepared as follows: Form 1 Sofosbuvir (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 became a sticky mass. No crystallization was observed. Approximately 10 mg of Form 7 [obtained from (i)] was added and the mixture was cooled to room temperature. The crystallization of Form 7 occurred during its storage overnight. A small amount of sticky mass remained on the stir bar, which was removed mechanically. The crystalline Form 7 was isolated by filtration and dried in a desiccator evacuated on silica gel.

* C Preparation of MTBE saturated in water (methyl fer-butyl ether)

MTBE saturated in water was prepared by vigorous stirring (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 MTBE saturated in water (800 ml) at room temperature. The solution was filtered through a pleated filter. The seed crystals of Sofosbuvir Form 7 (150 mg) were added and the solution was stored for two days in a closed flask at room temperature without agitation. 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 agitation. 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 MTBE saturated in water (30 ml) at room temperature. The solution was filtered through a folded filter and magnetically stirred (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 in vacuo (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 MTBE saturated in water (600 ml) at room temperature. The solution was filtered through a pleated filter. The seed crystals of Sofosbuvir Form 6 (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. The dissolution of Form 7 and Sofosbuvir Form 6 was measured (in triplicate) in 50 mM phosphate buffer at 37 ° C (pH 6.8, 900 ml) using USP apparatus II, 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 model with Chemical Station Dissolution Software. 400 g of Form 7 and Form 6 of Sofosbuvir were used for the assay and the percentages are given with respect to 400 g of PFA. The results are represented graphically in Figure 13.

* Example 20: Solubility and Particle Size 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.

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

The results are shown in the following table:

Table: Particle sizes (pm) of Form 7 and Form 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, o1 M HC1, 20 mM sodium acetate / acetic acid [pH 4.5], 50 mM of KH ² PO ⁴ [pH 6.8]). A stir bar was added, the vial was fixed in a block heater at 37 ° C and the suspension was shaken at 250 rpm. After 15 min and 1 h, the samples were extracted through a PTFE filter of 0.2 μm. 100 pl of the clear filtrate was diluted with 900 pl of acetonitrile / water mixture (1: 1) and 1 pl thereof was analyzed by UHPLC / UV:

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 The particle size of the examined batch of Form 7 is distinctly larger than that of Form 6. Therefore, Form 7 provides a significant advantage with respect to its solubility behavior.

* Example 21: Single glass data from Sofosbuvir Form 7

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

A small prism was selected as an appropriate identical crystal, since a sufficient volume of crystal could not be found. The data was collected on a Gemini Atlas CCD diffractometer (Cu Ka) using u> scans. The data collection runs were optimized for an absolute configuration analysis. The positional and anisotropic thermal parameters of all the non-hydrogen atoms were refined. H atoms bonded to carbon were placed geometrically. The positions of the H atoms were refined with slight restrictions in the lengths of union and angles to regularize their geometry (in the interval of CH 0.93-0.98). The positions of the hydrogen atoms bound to nitrogen and oxygen were initially refined with restricted binding lengths and then with conduction limitations. Uiso (H) was established in the range of 1.2-1.5 times Uec of the precursor atom. The contributions of component twin 1, component twin 2, component inverted 1 and component inverted 2 were refined to 0.60 (4), 0.42 (3), 0.00 (4) and 0.03 (3), therefore, the assignment of the absolute configuration is correct. Accessible voids of residual solvent were not 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 structure and absolute configuration analysis: Crystals [Betteridge, PW, Carruthers, JR, Cooper, RI, Prout, K. & Watkin, DJ (2003). J. Appl. Cryst. 36, 1487]; Molecular graph: Mercury; Vacuum calculation was performed using PLaton [Spek, AL (2003). PLATON, A multi-purpose crystallographic tool, University of Utrecht, Utrecht, The Netherlands]

Example 22- Pharmaceutical compositions (e.g., tablet) of sofosbuvir

Preparation method:

The batches were prepared by a dry granulation method. The intragranular ingredients of 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 obtained mixture was granulated by roller compactor and ground through a 0.8 mm screen and mixed with Part II of extragranular excipients for 10 minutes. Magnesium Stearate from 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 ratio is often used as an indication of the fluidity of a powder. The Hausner factor is the ratio of volume in mass to volume compacted, calculated by the formula density in volume / density compressed. 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 a poor flow.

Claims (15)

1. A process for preparing the crystalline E form of sofosbuvir comprising: (a) combining sofosbuvir with a solvent system comprising an organic solvent and optionally water, wherein the organic solvent is selected from the group consisting of aliphatic ethers - preferably C4-C8 ethers, cyclic ethers, ketones - preferably C3-C8 ketones , alcohols - preferably C4-C8 alcohols and, preferably wherein the organic solvent is selected from the group consisting of methyl t-butyl ether, t-butanol, acetone, methyl isobutyl ketone and, preferably, methyl isobutyl ketone; (b) optionally seeding with crystalline E-form; (c) adding an anti-solvent; (d) optionally isolating the crystalline E form; Y (e) optionally deagglomerating or triturating or micronising to obtain the crystalline E form of sofosbuvir having a smaller particle size; wherein the crystalline E form of sofosbuvir is 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 absence of peaks at: 10 , 9 and 14.2 degrees two theta ± 0.2 degrees two theta, and combinations of these data. 2. A process according to claim 1, wherein the crystalline E-form is filtered and dried. 3. A process according to any one of claims 1-2, wherein the solvent in step (c) is selected from the group consisting of aliphatic ethers - preferably C4-C8 ethers, alkanes - preferably C3-C8 alkanes and cycloalkanes, and preferably wherein the anti-solvent is selected from the group consisting of diisopropyl ether, n-heptane and cyclohexane, and preferably, cyclohexane. 4. A process according to any of claims 1-3, wherein step (a) is carried out in a solvent system comprising an organic solvent and water, preferably, wherein the amount of water is in the itnervalo 0% -2% by volume, or where the amount of water is in the range of 0.5 2%, or where the amount of water is about 1% by volume. 5. A process according to any of claims 1-4, wherein the reaction mixture of step (a) includes from about 10% to about 50% Sofosbuvir, by volume. 6. A process according to any of claims 1-5, wherein the process is carried out at temperatures between 0 ° C to 20 ° C, more preferably between 0 ° C to 15 ° C and most preferably between 6 to 12 ° C. A process according to any of claims 1-6, wherein the crystalline E form of Sofosbuvir is isolated by filtering the suspension or decanting the solvent from the suspension and comprising, additionally, washing and drying the crystalline Form E of sofosbuvir , preferably 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. 8. A process according to any of claims 1-7, further comprising a step of grinding, micronization or deagglomeration. 9. A process for preparing the crystalline E form of sofosbuvir according to claim 1, comprising: (TO) : (a) combine Sofosbuvir, preferably crystalline form 1 of sofosbuvir, with isobutyl methyl ketone and water, (b) seed with crystalline E form of sofosbuvir, (c) add cyclohexane, (d) optionally isolate form E and (e) optionally deagglomerating, triturating or micronising to obtain the crystalline E form of sofosbuvir having a desired particle size; or (B): (a) combine Sofosbuvir with isobutyl methyl ketone and water, where the amount of water is 1% by volume and the temperature is 9 ± 3 ° C, (b) sow with crystalline E form of sofosbuvir, (c) add cyclohexane, (d) isolate form E and (e) optionally micronizing. 10. A process according to claim 9, option (A), wherein the amount of water in step (a) is in the range of 0.5-2% by volume, and the mixture of preferably, wherein the amount of water is 1% by volume and the reaction is stirred at 50-200 RPM. 11. A process according to any of claims 9-10, wherein step (a) is carried out at a temperature ranging from 0 to 20 ° C, wherein stage (a) is carried out at 9 ± 3 °. C. 12. Form E of sofosbuvir prepared by a process according to any of claims 1 11. 13. The crystalline E form of micronized sofosbuvir or small particle size, where the crystalline E form of sofosbuvir of small particle size or micronized: (a) has a particle size distribution (DTP) of d (0.1) 0.4-0.6 micrometers, d (0.5) 1.0-2.0 micrometers and d (0.9) 3 2-5.0 micrometers or (b) shows a flow rate of not more than 100 s / 100 g, preferably not more than 80 s / 100 g, more preferably not more than 65 s / 100 g, preferably as measured by a test system of Automatic powder flow to a hole in Pharma Test 41-03300, or (c) shows a constant density of not less than 0.4 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, or (d) shows a constant density of not less than 0.4 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, or (e) is characterized by a water sorption of not more than 0.5%, more preferably not more than 0.3%, as measured by dynamic vapor sorption and water content after equilibration at 90% RH , or (f) any one or any combination of the properties of (a) to (e); or (g) a flow rate as defined in (b), a constant density as defined in (d) and a water sorption as defined in (e) or (h) a flow rate of not 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 not more than 0.3% as measured by DVS and water content after equilibration at 90% RH; Y wherein the crystalline E form of sofosbuvir is 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 absence of peaks at: 10 , 9 and 14.2 degrees two theta ± 0.2 degrees two theta, and combinations of these data. 14. A pharmaceutical composition comprising crystalline form E of micronized sofosbuvir or small particle size according to claim 13. Either sofosbuvir form E as defined in any of claims 12-13 or a pharmaceutical composition according to claim 14 for use as a medicament, preferably for the treatment of hepatitis C.