Synthetic process for an ecteinascidin

Abstract

The invention relates to a synthetic process for the synthesis of an ecteinascidin of formula (I) and to solid state forms of such ecteinascidin that are useful for the manufacture of medicaments.

Description

SYNTHETIC PROCESS FOR AN ECTEINASCIDINFIELD OF THE INVENTION

[0001] The present invention relates to the field of organic chemical synthesis, in particular to synthetic processes for the manufacture of an ecteinascidin compound and to solid-state forms of such ecteinascidin useful for the manufacture of a medicament.BACKGROUND OF THE INVENTION

[0002] International patent application publication WO 2018 / 197663 A1 discloses several synthetic analogs of ecteinascidin-736, a natural product isolated from Ecteinascidia turbinata. One of these compounds, of formula:is currently under clinical trials for the treatment of cancer under the international non-proprietary name (INN) ecubectedin.

[0003] As many ecteinascidins, it is obtained by a Pictet-Spengler reaction between compound 1 and tryptamine 2 in presence of acetic acid, followed by a CN-OH exchange promoted by silver salts according to Scheme I:3 X = CN -J AgNO3ecubectedin X = OHScheme I

[0004] WO 2018 / 197663 A1 also explored the synthesis and biological activity of a new kind of ecteinascidin analogs where a benzofuro[2,3-c]pyridine ring system is used.

[0005] Initial attempts to obtain one of such compounds, a compound of formula I:were unsuccessful because the Pictet-Spengler reaction between compound 1 and benzofuran 4 in acetic acid did not progress after adduct 5 was formed according to Scheme II:Scheme II

[0006] Finally, compound I was synthetized as described in WO 2018 / 197663 A1 by using cyanuric chloride (TCT) as the acid source and employing an excess of benzofuran 4 to give compound 6, followed by a CN-OH exchange promoted by silver salts according to Scheme III:Scheme III

[0007] The form of compound I obtained by the process described in WO 2018 / 197663 A1 , in the following named form A of compound I, is amorphous.

[0008] Although this process provides compounds 6 and I in a reproducible way, it presents several drawbacks such as the formation of a number of impurities including significant amounts of compounds II and III, resulting from the reaction between benzofuran 4 and TCT.

[0009] The formation of compounds II and III not only reduces the amount of benzofuran 4 available to react with compound 1 but also requires a specific chromatographic step to remove them. Benzofuran 4 is a valuable intermediate whose manufacture requires a multistep process and a resolution step as described in WO 2018 / 197663 A1 .

[0010] Compound I is currently under Phase I clinical trials for the treatment of cancer (NCT05841563).

[0011] Pharmaceutical-grade active substances must comply a number of quality requirements such a having a high purity and a controlled related substances profile.The need remains for an improved synthesis of compound I that reduces the excess of benzofuran 4 required and maintains the reaction yield when the manufacture scale is increased and provides compound I with higher purity and / or a simplified impurity profile.SUMMARY OF THE INVENTION

[0012] In a first aspect of the present invention there is provided a synthetic process, the process comprising reacting compound 1 with benzofuran 4 in presence of trifluoroacetic acid to give compound 6.

[0013] In a second aspect there is provided a synthetic process for the synthesis of compound I or a pharmaceutically acceptable salt thereof, the process comprising reacting compound 6 with water in presence of AgNOa6 I wherein the number of equivalents of AgNOs is from about 6 to about 15 and compound 6 is dissolved in a mixture of acetonitrile / water at an initial concentration from about 40 g / L to about 120 g / L.

[0014] In a third aspect, there is provided a method for reducing residual silver in compound I, the process comprising: a) dissolving compound I in a pharmaceutically acceptable, water-immiscible, polar organic solvent or a mixture of pharmaceutically acceptable, water-immiscible, polar organic solvents; b) washing such solution with an aqueous solution of a bromide or iodide salt, or mixtures thereof. c) decanting the organic phase, drying and filtering it; and d) evaporating the solvent to dryness.

[0015] In a fourth aspect, there is provided a method for crystallization of compound I using the addition of an antisolvent to obtain the desired crystal morphology and the voluminous physical properties necessary for a successful drug preparation process.

[0016] The process for the crystallization of compound I comprises the following steps: a) dissolving I in CH3CN at a temperature from about 75 °C to about 85 °C wherein the concentration of I is from about 0.020 and about 0.025 g / mL; b) cooling the solution to a temperature of from about 20 °C to about 30 °C; c) adding water until a dispersion of particles that does not re-dissolve is observed; d) maintaining the mixture at a temperature range of from about 0 °C to about 5 °C for a period of from about 15 to about 30 hours; wherein steps a) to d) are carried out with stirring.

[0017] In a fifth aspect, there is provided form B of compound I.

[0018] In a further aspect, there is provided compound I having a silver residual content of less than 1400 ppm.

[0019] In a further aspect, there is provided a pharmaceutical composition comprising form B of compound I or manufactured via form B of compound I.

[0020] In a further aspect, there is provided a process for the manufacture of pharmaceutical compositions comprising compound I, said process employing form B of compound I, preferably as starting material.

[0021] In a further aspect, there is provided form B of compound I for use in the manufacture of a pharmaceutical composition comprising compound I.

[0022] In a further aspect, there is provided the use of form B of compound I in the manufacture of a pharmaceutical composition comprising compound I.

[0023] In another aspect, there is provided form B of compound I for use as a medicament.

[0024] In another aspect, there is provided form B of compound I for use as a medicament for the treatment of cancer.

[0025] In another aspect, there is provided a method of treating an individual affected by cancer comprising administering to said affected individual a therapeutically effective amount of form B of compound I, or a therapeutically affective amount of compound I which has been manufactured via form B of compound I.

[0026] In another aspect, there is provided compound I comprising at least a detectible amount of form B, up to 1 % w / w form B, up to 5% w / w form B, up to 10% w / w form B, up to 50% w / w form B, up to 90% w / w form B, or being substantially pure form B.

[0027] In another aspect, there is provided a pharmaceutical composition comprising compound I comprising at least a detectible amount of form B, up to 1% w / w form B, up to 5% w / w form B, up to 10% w / w form B, up to 50% w / w form B, up to 90% w / w form B, or being substantially pure form B.BRIEF DESCRIPTION OF THE FIGURESFigure 1. Chromatographic profile of crude compound 6 obtained by the process described in WO2018 / 197663 A1.Figure 2. Chromatographic profile of crude compound 6 obtained by the synthetic process of this patent application.Figure 3. Chromatographic profile of purified compound 6 obtained by the process described in WO2018 / 197663 A1.Figure 4. Chromatographic profile of purified compound 6 obtained by the synthetic process described in this patent application.Figure 5 (a). X-ray powder diffractogram (XRPD) of form B of compound I. (b) enlarged section of the same diffractogram.DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

[0028] The term “related substance” refers to impurities and degradation products of Compound I.

[0029] The term “pharmaceutically acceptable salt” refers to any pharmaceutically acceptable salt which, upon administration to the patient is capable of providing (directly or indirectly) a compound as described herein. However, it will be appreciated that non-pharmaceutically acceptable salts also fall within the scope of the invention since those may be useful in the preparation of pharmaceutically acceptable salts. The preparation of salts can be carried out by methods known in the art.

[0030] For instance, pharmaceutically acceptable salts of the compounds provided herein are synthesized from the parent compound, which contain a basic or acidic moiety, by conventional chemical methods. Generally, such salts are, for example, prepared by reacting the free acid or base of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of both. Generally, nonaqueous media like ether, ethyl acetate, ethanol, 2-propanol or acetonitrile are preferred. Examples of acid addition salts include mineral acid addition salts such as, for example, hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate, and organic acid additions salts such as, for example, acetate, trifluoroacetate, maleate, fumarate, citrate, succinate, tartrate, malate, mandelate, benzenesulfonate, camphorsulfonate, sterylsulfate, 4-chlorobenzenesulfonate, 6,7- dihydroxycoumarin-4-methanosulfonate, 1 ,2-ethanedisulfonate, laurylsulfate, ethanesulfonate, 1 ,5-naphthalenedisulfonate, methanesulfonate and p-toluenesulfonate. Examples of the alkali addition salts include inorganic salts such as, for example, sodium, potassium, calcium and ammonium salts, and organic alkali salts such as, for example, ethylenediamine, ethanolamine, A / ,A / -dialkylenethanolamine, triethanolamine and basic aminoacids salts.

[0031] Pharmaceutically acceptable solvents in the context of the present invention are those classified under classes 2 and 3 of the guideline “Impurities: Guideline for residual solvents Q3C(R6) of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use.

[0032] The calculation of chromatographic content of compound I and any individual related substance, either as % area, % (w / w) or as % (respect to label), is carried out by HPLC using a DAD detector at / . = 254 nm (Bandwidth: 16 nm) and Reference off or using a UV detector at / . = 254 nm.

[0033] The calculation of the % area of compound I or any of its related substances is performed using the following formula:Aj x 100where

[0034] The calculation of the % w / w of compound I and each of its related substances in the drug substance is performed using the following formula:where

[0035] The calculation of the % w / w of compound I and of each of its related substances in the drug product is performed using the following formula:ATx VTx PRSx WR% (w / w) =ARx VRx F where

[0036] The concentration / content of compound I in the bulk solution is calculated using the following equationsATx VFTx WRx PRSConcentration (ma / J) = - ARX VBX VRX 100% (respect to label') = Concentration (mg / mL) x 100 where

[0037] Total related substances should be calculated as the sum of reported individual impurities.

[0038] For measuring chromatographic content of compound I or of any related substance in samples containing additional materials, such as pharmaceutical excipients, peaks arising from the additional materials are discarded. Additionally, peaks from a blank chromatogram, also called system peaks, are also discarded. In order to discard these peaks, blank and placebo chromatograms are obtained in addition to the test solution chromatogram.

[0039] The calculation of % w / w in mixtures of solid-state forms of compound I is carried out by collecting powder X-ray data from essentially homogeneous samples of known forms of compound I and from the problem sample. X-ray data from known forms is used to identify distinguishing peaks for each polymorph. The relative amount of each polymorph in the problem sample can be determined by integrating peak heights or peak areas of reflections that are unique to each compound I polymorph in the sample, according to standard quantitative X-ray methods, with integration of peak areas generally been preferred.

[0040] Typically, the XRPD analysis can be simplified by visually inspecting the diffractogram to determine which polymorphs are present at detectable levels, and then limiting integration to the unique reflections corresponding to those polymorphs.

[0041] To provide a more concise description, some of the quantitative expressions given herein are not qualified with the term “about”. It is understood that, whether the term “about” is used explicitly or not, every quantity given herein is meant to refer to the actual given value, and it is also meant to refer to the approximation to such given value that would reasonably be inferredbased on the ordinary skill in the art, including equivalents and approximations due to the experimental and / or measurement conditions for such given value.

[0042] In a first aspect of the present invention, there is provided a synthetic process, the process comprising reacting compound 1 with benzofuran 4 in presence of trifluoroacetic acid to give compound 6.Preferred embodiments for the first aspect of the present invention:

[0043] In a preferred embodiment, the process is carried out in neat TFA or in a solvent selected from acetonitrile, toluene, isopropanol, hexafluoroisopropanol, and ethanol. Preferably, the reaction is carried out in a solvent selected from acetonitrile and toluene, being acetonitrile the most preferred solvent.

[0044] In another preferred embodiment, the process is carried out under water-removing conditions. Examples of water-removing conditions include, but are not limited to, azeotropic removal of water and water adsorption with molecular sieves.

[0045] In another preferred embodiment, the process is carried out using between about 2 and about 10 equivalents of trifluoroacetic acid, preferably between about 2 and about 5 equivalents of trifluoroacetic acid, more preferably between about 2.5 and about 4 equivalents of trifluoroacetic acid, being most preferred about 3 equivalents of trifluoroacetic acid.

[0046] In another preferred embodiment, the initial concentration of compound 1 is from about 40 g / L to about 80 g / L, being more preferred from about 50 g / L to about 70 g / L. Even more preferably from about 60 g / L to about 65 g / L. Most preferred initial concentration of compound 1 is about 62.5 g / L.

[0047] In another preferred embodiment, the process is carried out at a temperature of from about 80 °C to about 90 °C, being more preferred from about 83 °C to about 87 °C. Most preferred reaction temperature is about 85 °C.

[0048] In another preferred embodiment, the reaction time for the synthesis of compound 6 is between about 8 to about 24 hours, preferably between about 12 and about 20 hours and most preferably about 16 hours.

[0049] In another preferred embodiment, the process is carried out using from about 1 .5 to about 5 equivalents of benzofuran 4, being more preferred using between about 1.5 and about 3 equivalents of benzofuran 4. It is most preferred to use about 2 equivalents of benzofuran 4.

[0050] In a preferred embodiment, the reaction mixture was quenched at ambient temperature by adding an aqueous NH4OH / NH4CI buffer solution until the pH of the mixture is about 9.

[0051] In a preferred embodiment, crude compound 6 is purified by flash chromatography under silica gel using a mixture of hexane:ethyl acetate:dichloromethane:methanol in a 6:2:2:0.1 v / v ratio as eluent.

[0052] In a second aspect, there is provided a synthetic process for the synthesis of compound I or a pharmaceutically acceptable salt thereof, the process comprising reacting compound 6 with water in presence of AgNOa6 I wherein the number of equivalents of AgNOs is from about 6 to about 15 and compound 6 is dissolved in a mixture of acetonitrile / water at an initial concentration from about 40 g / L to about 120 g / L.Preferred embodiments for the second aspect of the present invention:

[0053] In a preferred embodiment, compound 6 used for the manufacture of compound I is obtained following the procedure described in the first aspect of the present invention, optionally including one or more of its preferred embodiments.

[0054] In a preferred embodiment, the number of equivalents of AgNOs is from about 7 to about 14, being more preferred from about 8 to about 12 and even more preferred from about 9 to about 11. Most preferably, the number of equivalents of AgNOs is about 10.

[0055] In a preferred embodiment, the v / v ratio acetonitrile / water is from about 3:1 to about 1 :1 , more preferably from about 2:1 to about 1 :1 , even more preferably from about 1 .5:1 to about 1 .3:1 and most preferably about 1 .4:1 .

[0056] In a preferred embodiment, the initial concentration of compound 6 in the acetonitrile / water mixture is from about 60 g / L to about 100 g / L, being more preferred from about 70 g / L to about 90 g / L and even more preferred from about 75 g / L to about 80 g / L. Most preferably, the initial concentration of compound 6 is about 79 g / L.

[0057] In a preferred embodiment, crude compound I is purified by chromatography under silica gel using as eluent a gradient of CH2Cl2:MeOH from 99:1 to 94:6.Preferred embodiments of the process for reducing residual silver in compound I:

[0058] In a preferred embodiment, compound I used in this purification method is prepared according to the method defined in the second aspect of the invention, optionally including one or more of its preferred embodiments.

[0059] In a more preferred embodiment, compound I used in this purification method is prepared according to the methods defined in the first and the second aspects of the present invention, optionally including one or more of their preferred embodiments.

[0060] Examples of pharmaceutically acceptable, water immiscible, polar solvents suitable for this step are chloroform, 1 -butanol, 2-butanol, butyl acetate, ethyl acetate, methyl acetate, 1- pentanol, propyl acetate and dichloromethane or mixtures thereof. More preferred pharmaceutically acceptable, water-immiscible, polar solvents for this step are chloroform, ethyl acetate and dichloromethane or mixtures thereof. Most preferably, the pharmaceutically acceptable, water immiscible, polar solvent is dichloromethane.

[0061] Examples of bromide and iodide salts are NaBr, Nal, KBr, KI, MgBr2, Mgh, CaBr2, Cab, ammonium bromide, ammonium iodide and mono-, di-, tri- and tetraalkyl ammonium bromide or iodide. Preferably silver is removed by washing with an aqueous solution of an iodide salt, more preferably Nal or KI, most preferably Nal.

[0062] In a preferred embodiment, the concentration of the bromide or iodide salt in the aqueous solution is from about 5% w / w to the saturation concentration of such agent in water, being more preferred from about 10% to about 30% w / w and most preferably about 20% w / w.

[0063] In a preferred embodiment, the washing step is repeated from about 2 to about 10 times, being more preferred from about 3 to about 8 times, being more preferred from about 3 to about 6 times, being most preferred from 4 to 5 times.

[0064] In a preferred embodiment, the process for reducing residual silver in compound I is carried out after the chromatographic purification of compound I.

[0065] In a preferred embodiment, the compound I obtained after the chromatographic purification is dissolved in dichloromethane and washed repeatedly with aqueous 20% w / w sodium iodide, being preferred repeating the washing step between about 1 to about 10 times, more preferably between about 3 to about 8 times and most preferably about 4-5 times.Preferred embodiments of the process for the crystallization of compound I:

[0066] In a preferred embodiment, compound I used in this crystallization process has been previously purified as defined in the third aspect of the present invention, optionally including one or more of its preferred embodiments.

[0067] In a preferred embodiment, compound I used in this crystallization process has been obtained as defined in the second aspect of the present invention, optionally including one or more of its preferred embodiments.

[0068] In a preferred embodiment, compound I used in this crystallization process has been obtained as defined in the second aspect of the present invention and has been previously purified as defined in the third aspect of the present invention, optionally including one or more of their preferred embodiments.

[0069] In a preferred embodiment, the concentration of compound I in the CH3CN solution of step a) is about 0.022 g / mL.

[0070] In a preferred embodiment, the temperature of step a) is about 82 °C.

[0071] In a preferred embodiment, stirring is carried out by mechanical or magnetic means.

[0072] The process involves adding water in step c) until a dispersion of particles that does not re-dissolve is observed. By “dispersion of particles that does not re-dissolve is observed” it is understood to mean an amount of water is added such that a dispersion of particles is observed in the solution that does not disappear with stirring. Having carried out the process, it will be possible to determine how much water will achieve the dispersion of particles that does not redissolve for a given amount of compound I and given solution volume. Once the amount of water is known, it may be possible to proceed to the next step without observing the solution to confirm the presence of the dispersion.

[0073] In a preferred embodiment, the amount of water added in step c) is from about 25 and about 35 mL / g of compound I, more preferably about 30 mL / g of compound I.

[0074] In a preferred embodiment, the mixture of step d) is maintained at a temperature range of from about 0 °C to about 5 °C for about 20 to about 24 hours.

[0075] In a preferred embodiment, the crystallization process further comprises: e) separating crystalline compound I; f) washing the crystalline material with water; and g) drying the crystalline compound I under vacuum at a temperature of from about 25 °C to about 30 °C.

[0076] The obtained crystalline form of compound I can be separated by isolation operations such as filtration or centrifugation, preferably by filtration.

[0077] In a preferred embodiment, drying step g) is carried out under vacuum where the resulting pressure is lower than about 10 mbar, preferably lower than 5 mbar.

[0078] In a preferred embodiment, drying step g) is carried out at a temperature of about 27 °C.

[0079] In a preferred embodiment, drying step g) last from about 36 to about 72 hours, more preferably about 48 h.Preferred embodiments related to Form B of compound I

[0080] Form B of compound I can be characterized by showing an X-ray powder diffractogram pattern using Cu-Kaiand Cu-Kce radiation comprising three of more characteristic peaks at 2- theta angles selected from 9.5 ± 0.2°, 12.8 ± 0.2°, 14.8 ± 0.2°, 15.4 ± 0.2°, 16.2 ± 0.2° and 22.0 ± 0.2°. Form B may alternatively be characterized by showing an X-ray powder diffractogram pattern using Cu-Kai and Cu-Kce radiation comprising four or more of said characteristic peaks. Alternatively, form B may be characterized by showing an X-ray powder diffractogram pattern using Cu-Kai and Cu-Kce radiation with five or more of said characteristic peaks. Alternatively, form B may be characterized by showing an X-ray powder diffractogram pattern using Cu-Kai and Cu-Ka2 radiation comprising all six of said characteristic peaks.

[0081] Particularly, form B of compound I can be characterized by an X-ray powder diffractogram pattern using Cu-Kai and Cu-Ka2 radiation comprising peaks and intensities as shown in the following table:

[0082] In a preferred embodiment, further peaks may be found at 2-theta angles of 10.0 ± 0.2°, 19.2 ± 0.2°, 19.9 ± 0.2°, and 21.2 ± 0.2°. Particularly, form B of compound I can be characterizedby an X-ray powder diffractogram pattern using Cu-Kaiand Cu-Kce radiation comprising characteristic peaks and intensities as shown in the following table:

[0083] In a more preferred embodiment, further peaks may be found at 2-theta angles of 7.0 ± 0.2° and 7.5 ± 0.2°. Particularly, form B of compound I can be characterized by an X-ray powder diffractogram pattern using Cu-Kai and Cu-Ka2 radiation comprising characteristic peaks and intensities as shown in the following table:

[0084] In a most preferred embodiment, the present invention relates to form B of compound I that exhibits an X-ray powder diffraction pattern using Cu-Kai and Cu-Kce radiation substantially the same as the X-ray powder diffraction patterns depicted in Figure 5.

[0085] In a further aspect, there is provided compound I having a residual silver content of less than 1400 ppm.

[0086] In a preferred embodiment, the residual content is of less than 1000 ppm, preferably less than 500 ppm, more preferably less than 100 ppm.Pharmaceutical compositions and processes for their manufacture

[0087] Pharmaceutical compositions of compound I that can be used include solutions, lyophilized compositions, etc., with suitable excipients for intravenous administration.

[0088] In embodiments according to the present invention the pharmaceutical composition of compound I is a ready-to-use injectable compound I solution, which is preferably easy to administer without need for reconstitution.

[0089] In some embodiments, there is provided a storage-stable, ready-to-use, injectable liquid parenteral composition including compound I and one or more pharmaceutically acceptable solvents, co-solvents and / or solubilizing agents.

[0090] In some embodiments, the pharmaceutically acceptable solvent is selected from ethanol, polyethylene glycol and propylene glycol.

[0091] In some embodiments, the ready-to-use injectable compound I solution further includes a pharmaceutically acceptable stabilizing agent. Preferably, the stabilizing agent is an aminoacid selected from glycine, arginine, histidine, alanine, isoleucine, leucine, asparagine, lysine, aspartic acid, methionine, cysteine, phenylalanine, glutamic acid, threonine, glutamine, tryptophan, valine, ornithine, proline, selenocysteine, serine, tyrosine, citrulline, preferably glycine or L-arginine.

[0092] In some embodiments, the ready-to-use injectable compound I solution further comprises a buffer.

[0093] In some embodiments, the pH of the ready-to-use injectable compound I solution is from about 2.5 to about 5.0.

[0094] In some embodiments, the ready-to-use injectable compound I solution further includes an antioxidant such a BHT, BHA or bisulfite.

[0095] In embodiments, compound I is supplied and stored as a stable and sterile lyophilized product comprising compound I, a buffer, a bulking agent and a sufficient amount of acid or base to provide an appropriate pH for injection when the composition is reconstituted in an appropriate solvent.

[0096] In embodiments according to the present invention, the pre-lyophilized compound I or pre-solution compound I comprises at least some crystalline material. The pre-lyophilized / presolution compound I may be crystalline. The pre-lyophilized / pre-solution compound I may be a solid-state crystalline form as described herein. The pre-lyophilized / pre-solution of compound I may contain form B as described herein. Using compound I as defined in the present invention leads to advantages, for example better control of related substances.

[0097] As such, the present invention provides pharmaceutical compositions and their methods of preparation wherein the composition manufacturing process utilized a solid-state form as disclosed herein. In an embodiment, this is form B.

[0098] In some embodiments, the buffer is an organic carboxylic acid buffer that is derived form an organic acid selected from the group consisting of lactic acid, butyric acid, propionic acid, acetic acid, chloroacetic acid, succinic acid, citric acid, ascorbic acid, tartaric acid, malic acid, maleic acid, fumaric acid, glutamic acid, aspartic acid, gluconic acid, glycine, histidine, phthalic acid, and o-ketoglutaric acid. In some embodiments, the organic carboxylic acid buffer is derived form an organic acid selected from lactic acid or succinic acid. In some embodiments, the organic acid carboxylic acid buffer is derived from histidine or glycine. In some embodiments, the organic carboxylic acid buffer is derived from lactic acid.

[0099] In some embodiments, the inorganic acid buffer is derived from an inorganic acid selected from the group consisting of phosphoric acid and hydrochloric acid. In some embodiments, the inorganic acid buffer is phosphate buffer prepared with potassium dihydrogen phosphate and phosphoric acid with potassium hydroxide to adjust the pH as required.

[0100] In some embodiments, the bulking agent is selected from aminoacids, cyclodextrins, polyols, disaccharides and high polymer materials; or mixtures thereof. In some embodiments the bulking agent is a disaccharide.

[0101] In some embodiments, the aminoacid is selected from the group consisting of glycine, arginine, histidine, alanine, isoleucine, leucine, asparagine, lysine, aspartic acid, methionine, cysteine, acetylcysteine, selenocysteine, phenylalanine, glutamic acid, threonine, glutamine, tryptophan, valine, ornithine, proline, serine and tyrosine. In some embodiments, the aminoacid is L-arginine.

[0102] In some embodiments, the cyclodextrin is selected from the group consisting of hydroxypropyl-p-cyclodextrin, hydroxyethyl-p-cyclodextrin, 2-sulfobutyl-p-cyclodextrin, methylated-p-cyclodextrin, glucosyl-p-cyclodextrin, maltosyl-p-cyclodextrin and carboxymethyl-p- cyclodextrin. In some embodiments, the cyclodextrin is hydroxypropyl-p-cyclodextrin.

[0103] In some embodiments, the polyol is selected from the group consisting of sorbitol, mannitol, glycerol, and lactitol. In some embodiments, the polyol is mannitol.

[0104] In some embodiments, the disaccharide is selected form the group consisting of sucrose, trehalose or lactose, or a combination thereof. In some embodiments, the disaccharide is sucrose.

[0105] In some embodiments, the high polymer material is selected from the group consisting of albumin, dextran, hydroxyethyl starch, sodium carboxymethyl cellulose, maltodextrin, polyvinylpyrrolidone (PVP) and polyethylene glycol. In some embodiments, the high polymer material is dextran 40.

[0106] In some embodiments, the base is selected from the group consisting of carbonates, hydroxides, hydrogen carbonates and ammonium salts. Particularly preferred bases are sodium carbonate, potassium carbonate, calcium carbonate, NH4OH, sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium hydrogen carbonate, potassium hydrogen carbonate and calcium hydrogen carbonate. In some embodiments, the base is potassium hydroxide.

[0107] In some embodiments, the pH of the reconstituted lyophilized composition is about 3. In some embodiments, the pH of the reconstituted lyophilized composition is from about 2.5 to about 5. In some embodiments, the pH of the reconstituted lyophilized composition is from about 2.5 to about 3.5. In some embodiments, the pH of the reconstituted lyophilized composition is 2.8 to 3.3.

[0108] In some embodiments, the stable lyophilized product comprises compound I, phosphate and sucrose and the pH of the reconstituted lyophilized composition is 2.8 to 3.3. In some embodiments, the stable lyophilized product comprises about 3 mg compound I, about 10.2 mg of phosphate, wherein said phosphate is calculated as potassium dihydrogen phosphate, and 300 mg of sucrose.

[0109] The compound I containing formulations of this invention can be made by freeze-frying a composition of this invention in the form of a buffered bulk solution including compound I, a buffer, and a disaccharide. The disaccharide is preferably sucrose. Usually, the bulk solution will be buffered, for example to a pH of about 2.5 to 5, preferably about 2.5 to 3.5, more preferably pH 2.8 to 3.3. The preferred buffering agent is phosphate buffer. In preferred embodiments, the phosphate buffer comprises potassium dihydrogen phosphate and phosphoric acid with potassium hydroxide to adjust the pH, if needed.

[0110] As such, in embodiments of the present invention there is provided a buffered lyophilized composition comprising compound I, a buffer such as phosphate buffer and a disaccharide; wherein the buffer is configured such that upon reconstitution the pH of the reconstituted lyophilized composition is from about 2.5 to about 5, preferably from about 2.5 to about 3.5, more preferably 2.8 to 3.3.

[0111] The present invention has identified methodologies that allow for complete dissolution of compound I in desired buffers whilst minimizing impurity generation. In embodiments, the use of phosphoric acid allows for direct dissolution of compound I, preferably at pH about 1 to about 2, followed by addition of water for injection, potassium dihydrogen phosphate (preferably to have a pH about 2.5 to 5, preferably 2.5 to 3.5, most preferably 2.8 to 3.3), bulking agent such as disaccharide, preferably sucrose. Such a formulation strategy enables a rapid dissolution into the bulk formulation limiting the formation of impurity H during the manufacture. In an embodiment, there is provided direct dissolution of compound I, comprising dissolving compound I in an inorganic buffer (preferably at pH about 1 to 5, about 2.5 to 5, about 2.5 to 3.5 or about 2.8 to about 3.3), followed by addition of bulking agent such as disaccharide, preferably sucrose, to form a bulk solution. The bulk solution may undergo sterilizing filtration. The bulk solution may then be filled in vials according to the desired dose. The bulk solution in vials may be lyophilized to form a lyophilized buffered compound I formulation. The lyophilized formulation may then be reconstituted to form a reconstituted solution. The reconstituted solution may be diluted to form an injection solution. Preferably, with direct dissolution the compound I is amorphous or substantially amorphous.

[0112] In an embodiment the disaccharide is previously dissolved in acidified water.

[0113] In an embodiment, the pH of the injection solution is from about 3 to about 5.

[0114] As explained herein, compound I has limited aqueous solubility. It was found that compound I solubility is improved in the bulk solution by first forming a concentrated pre-solution of compound I in an inorganic acid, for example phosphoric acid, which is further diluted with water for injection. A basic ingredient, such as potassium dihydrogen phosphate, is then added followed by a disaccharide such as sucrose and upon adjusting the pH to a set value of between about 2.8 to about 3.3 to obtain the compound I bulk solution in a phosphate buffer, pH = 2.8-3.3, containing a disaccharide (for example, sucrose).

[0115] Provided are processes useful for improving the solubility of compound I in the bulking solution that comprise dissolving compound I in phosphoric acid, for example 0.1 N phosphoric acid and subsequent dilution of the solution with water for injection to yield a compound I concentrated solution, dissolving in such solution potassium dihydrogen phosphate and a disaccharide, and, optionally, adjusting the pH. In some illustrative, but not limiting embodiments of this invention, pH adjustment is accomplished with phosphoric acid or potassium hydroxide.

[0116] Illustrative embodiments of bulk solution for freeze-drying according to the present invention are provided by a solution of compound I buffered at pH 2.8-3.3 with phosphoric acid and potassium dihydrogen phosphate with sucrose as bulking agent.

[0117] An illustrative embodiment of the methodology according to this invention provides as follows: compound I is dissolved in 0.1 N phosphoric acid to yield a compound I solution in a ratio of 1 g compound I / 175 g phosphoric acid (0.1 N), and, after complete dissolution, subsequently diluted with water for injection (ca. 525 mL per gram of compound I). To the resulting solution, it is added potassium dihydrogen phosphate (3.4 g per gram of compound I) and sucrose (100 g per gram of compound I). Dissolution is visually checked at all steps before continuing, and dissolution is considered complete when it is so appreciated visually. The pH of the solution is checked and, if needed, adjusted to a value in the range from about 1 to about 5, more preferably in the range from about 2.5 to about 5, even more preferably in the range from about 2.5 to about 3.5, and most preferably in the range from about 2.8 to about 3.3 by slow addition of a suitable acid or base. A preferred embodiment of such acid is phosphoric acid, in which case a preferred concentration is about 0.1 N. A preferred embodiment of such base is potassium hydroxide, preferably in solution, in which case a preferred concentration is about 0.1 N. The volume is finally adjusted by addition of a suitable, biocompatible fluid, preferably water for injection. The bulk solution is then filed in vials according to the desired dose.

[0118] Compound I may be pre-dissolved in high concentration inorganic acid. In a preferred embodiment, the pre-dissolution step is at least 20 minutes, at least 30 minutes, at least 60 minutes or at least 90 minutes, between 20-90 minutes, between 30-90 minutes, between 20-40 minutes, between 20-30 minutes or around 20 minutes. Following dissolution, the pre-dissolution solution can be diluted to form the required concentration. Dilution may involve x1 , x2, x3 or moredilutions with water for injection to obtain the target concentration. In embodiments, dilutions are carried out to achieve the desired concentration at appropriate molarity.

[0119] During manufacture, reducing the dissolution time of compound I is advantageously achieved with limited phosphoric acid. Reducing the dissolution time results in lower levels of impurity H during manufacture. As such using high molarity phosphoric acid can achieve a high compound I concentration in a limited organic acid volume.

[0120] In embodiments, a multi-step compounding strategy is used to prepare compositions comprising compound I. Step 1 is the pre-dissolution step described above, for example: predissolving crystalline compound I in phosphoric acid 0.1 N and diluting with water for injection (WFI). The remaining excipients are added sequentially in solid form and stirred until total dissolution of each of them. The final bulk solution composition is obtained after adjustment of water for injection to final volume. The concentration of active compound in the final bulk solution is 1 mg / mL. The present invention therefore identifies a compounding strategy to formulate crystalline compound I.

[0121] In one embodiment, the lyophilized composition comprises or consists of 3 mg of active compound, 300 mg of sucrose, 10.2 mg of phosphate calculated as potassium dihydrogen phosphate. In some embodiments, the weight ratio in the lyophilized composition is between 0.9% and 1.1 % (w / w) of active compound, 95% to 97% (w / w) sucrose, 3% phosphate (w / w) calculated as potassium dihydrogen phosphate. In preferred embodiments, the weight ratio in the lyophilized composition is 1% (w / w) active compound, 96% (w / w) sucrose, 3% (w / w) phosphate calculated as potassium dihydrogen phosphate. When reconstituted to 10 mL in the vial the resulting solution has a pH of about 3 (range of pH 2.5 to 3.5, preferably 2.8 to 3.3).

[0122] The lyophilized material is usually present in a vial, which contains a specified amount of compound I. Preferably the lyophilized composition of compound is provided in a 10 mL vial. The specified amount of compound I in a lyophilized composition can be from 0.2 to 15 mg / vial, preferably from 1 to 10 mg / vial; or about 1 mg / vial, about 2 mg / vial, about 3 mg / vial, about 4 mg / vial, about 5 mg / vial, about 6 mg / vial, about 7 mg / vial, about 8 mg / vial, about 9 mg / vial or about 10 mg / vial. The specified amount of compound I in a lyophilized composition is preferably from about 3 mg / vial to about 10 mg / vial, being more preferred from about 3 mg / vial to about 7 mg / vial. In lyophilized embodiments, the composition contains between 0.9% w / w and 1.1 % w / w of compound I, preferably it is 1 % w / w.

[0123] It is necessary to ensure that the composition of compound I is sterile and is aseptically filled into vials. This is critical for parenteral drugs. According to embodiments of the present invention, terminal sterilization by heat or gamma irradiation are not used to avoid degradation of compound I. Instead, according to embodiments of the present invention, a sterilization filtration of the bulk compound I solution is carried out before aseptic vial filling. In embodiments, the filter may be filters such as PVDF or PES. In embodiments the filter may be a 0.2 pm filter.Storage of Pharmaceutical compound I Formulations

[0124] Embodiments of this invention also provide a method of storing a lyophilized compound I composition, wherein the lyophilized compound I composition is manufactured from a solid-state form as disclosed herein - form B. The use of solid-state forms of the present invention may lead to advantageous storage properties as further discussed below. As discussed herein, using compound I as defined in the present invention can lead to advantages, including better control of impurities and / or degradation products.

[0125] It is advantageous to ensure that compound I is stable during at least 24 months. The compound I lyophilized formulations are storage stable such that after prolonged storage a 5 °C ± 3 °C, the compound I retains its therapeutic effectiveness and exhibits minimal chemical degradation (e.g degradation is minimized and within acceptable tolerance; for example, the related substances profile of the compound I, amount of each related substance, compound I content, as determined by HPLC analysis, are substantially the same before and after prolonged storage).

[0126] In embodiments, the lyophilized compound I compositions of the present disclosure minimize the amount of a compound I degradation product resulting from deacetylation of compound I (Impurity H) when the composition is stored for prolonged times (e.g. at least 24 months). In some embodiments, the amount of impurity H present is less than 0.1 %, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1 .0% w / w of the total compound I weight in the formulation after prolonged storage at 5 °C ± 3 °C. In other embodiments, the amount of impurity H present is less than 0.1 %, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1.0% area in the formulation after prolonged storage at 5 °C ± 3 °C. Impurities B, D, E and H have the following structures:

[0127] Impurities B, D, E and F are formed during the manufacture of compound I. Impurity H was formed during the manufacture of the lyophilized composition and it is also a degradation product when the lyophilized composition is stored a 25 °C.

[0128] In a preferred embodiment the method of storing a lyophilized compound I composition comprises storing a lyophilized composition comprising 3 mg / vial compound I, phosphate bufferand a disaccharide at a temperature of 5 °C ± 3 °C for at least 24 months, wherein the lyophilized composition is formulated such that reconstitution with 10 mL of water for injection will yield a solution having a pH of 2.8 to 3.3 and a compound I concentration of 0.3 mg / mL and wherein after the at least 24 months storage, the amount of Impurity H present in the composition is not more than 1 .0% w / w of the total compound I weight or not more than 1 .0% area. In some embodiments, the lyophilized compound I composition is stored at a temperature of 5 °C ± 3 °C for, or for at least, 24 months, 30 months, 36 months, 42 months, 48 months or 60 months, wherein after 24 months, 30 months, 36 months, 42 months, 48 months or 60 months of storage, the amount of a compound I degradation product Impurity H present in the composition is not more than 1 .0% w / w of the total compound I weight or not more than 1 .0% area. In some embodiments, the amount of Impurity H present in the composition after storage at about 5 °C ± 3 °C for 60 months is not more than 1 .0% w / w, or is less than 0.9% w / w, less than 0.8% w / w, less than 0.7% w / w, less than 0.6% w / w, less than 0.5% w / w, less than 0.4% w / w, less than 0.3% w / w, less than 0.2% w / w or less than 0.1% w / w of the total compound I weight or it is not more than 1.0% area, or is less than 0.9% area, less than 0.8% area, less than 0.7% area, less than 0.6% area, less than 0.5% area, less than 0.4% area, less than 0.3% area, less than 0.2% area or less than 0.1 % area. In one embodiment, the amount of compound I degradation product Impurity H present in the composition is not more than 1 .0% w / w of the total compound I weight or not more than 1 .0% area after at least 36 months of storage. In some embodiments, the total related substances (as % area) after storage at about 5 °C ± 3 °C for 24 months, 30 months or 36 months is not more than 0.6%, 0.7%, 0.8%, 0.9% or 1.0% (% area). In some embodiments, the total related substances (as % w / w) after storage at about 5 °C ± 3 °C for 24 months, 30 months or 36 months is not more than 0.6%, 0.7%, 0.8%, 0.9% or 1.0% (% w / w). In some embodiments, the initial amount of Impurity H present in the composition (i.e. one day of lyophilization) is less than 0.4% w / w of the total compound I weight or less than 0.4% area. In some embodiments, the initial amount of Impurity H present in the composition is not more than 1 .0% w / w, not more than 0.9% w / w, not more than 0.8% w / w, not more than 0.7% w / w, not more than 0.6% w / w, not more than 0.5% w / w, not more that 0.4% w / w, not more than 0.3% w / w, not more than 0.2% w / w or not more than 0.1 % w / w. In some embodiments, the initial amount of Impurity H present in the composition is not more than 1 .0% area, not more than 0.9% area, not more than 0.8% area, not more than 0.7% area, not more than 0.6% area, not more than 0.5% area, not more than 0.4% area, not more than 0.3% area, not more than 0.2% area or not more than 0.1 % area.

[0129] In some embodiments, after storage at about 5 °C ± 3 °C for 24 months, 30 months, 36 months, 48 months or 60 months the stable, lyophilized, compound I formulation shows negligible degradation of compound I assay content, for example, a decrease within 1.5%, 1.0%, 0.5% or 0.2% (% respect to label) of the total amount of compound I in the bulk solution from which the formulation is made.

[0130] Accordingly, provided are stable, lyophilized compound I formulations comprising a buffer derived from an inorganic acid (e.g. phosphate buffer) at a molar ratio of buffer to compound I ofabout 20:1 , including the molar ratio 40:1 to 10:1 , 30:1 to 15:1 , and sucrose as a bulking agent, which, when reconstituted in 10 mL of water has a pH of about 3.0, including pH 2.5-3.5 or pH 2.8-3.3, which comprises Impurity H at no more than 1.0% w / w, or is less than 0.9% w / w, or is less than 0.8% w / w, or is less than 0.7% w / w, or is less than 0.6% w / w, less than 0.5% w / w, or is less than 0.4% w / w, or is less than 0.3% w / w, or is less than 0.2% w / w or less than 0.1 % w / w of the total weight of compound I and, preferably, Impurity H does not increase to more than 1.0% w / w of the total weight of compound I after storage at 5 °C ± 3 °C for 12 months, 24 months, 30 months, 36 months, 48 months or 60 months; or storage at 25 °C / 60% RH for 3 months, 6 months or 12 months. In these embodiments, the compound I is 95 to 105%, or 96 to 100% of 3-10 mg compound I or of the amount of compound I by assay at day 1.

[0131] In some embodiments, provided are stable, lyophilized compound I formulations comprising a buffer derived from an inorganic acid (e.g. phosphate buffer) at a molar ratio of buffer to compound I of about 20:1 , including the molar ratio 40:1 to 10:1 , 30:1 to 15:1 , and sucrose as bulking agent, which, when reconstituted in 10 mL of water has a pH of about 3.0, including pH 2.5-3.5 or pH 2.8-3.3, which comprises Impurity H at no more than 0.8% area, or is less than 0.7% area, or is less than 0.6% area, less than 0.5% area or less than 0.4% area and, preferably, Impurity H does not increase to more than 0.8% area after storage at 5 °C ± 3 °C for 12 months, 24 months, 30 months, 36 months, 48 months or 60 months; or storage at 25 °C / 60% RH for 3 months, 6 months or 12 months. In these embodiments, the compound I is 95 to 105%, or 96 to 100% of 3 mg compound I or of the amount of compound I by assay at day 1 .

[0132] Also provided are methods of reducing compound I degradation in a lyophilized formulation by incorporating a buffer derived from an inorganic acid, preferably a phosphate buffer, in the lyophilized formulation with the compound I such that the impurity H in the formulation does not exceed 0.1 % w / w, 0.2% w / w, 0.3% w / w, 0.4% w / w, 0.5% w / w, 0.6% w / w, 0.7% w / w, 0.8% w / w, 0.9% w / w or 1 .0% w / w of the total compound I weight or does not exceed 0.1 % area, 0.2% area, 0.3% area, 0.4% area, 0.5% area, 0.6% area, 0.7% area, 0.8% area, 0.9% area or 1.0% area after storage at 5 °C ± 3 °C for 12 months, 24 months, 30 months, 36 months, 48 months or 60 months; or storage at 25 °C / 60% RH for 1 month, 3 months, 6 months or 12 months, particularly when the amount of compound I is 95% to 105%, or 96% to 100% of 3 mg of compound I or of the amount of compound I by assay at day 1.

[0133] Other impurities or degradation products that may be minimized in the storage of the stable, lyophilized compound I formulation may be the degradation products with the following relative retention time on the commercial HPLC method: rrt 0.12, rrt 0.20-0.23, rrt 0.85, rrt 0.93- 0.94 and rrt 1.04 (Impurity D).

[0134] In further embodiments, the total residual water content for the lyophilized compound I formulation is not more than 3% (w / w), preferably not more than 2.5% (w / w).

[0135] Embodiments of the invention further provide a pharmaceutical product comprising a vial containing a lyophilized compound I composition. In a preferred embodiment, the pharmaceutical product comprises a vial containing a lyophilized composition consisting of from about 3 to about 10 mg compound I, from about 10.2 to about 33.4 mg phosphate calculated as potassium dihydrogen phosphate and from about 300 to about 1000 mg sucrose and a label affixed to the vial comprising an expiration date at least 48 months from the date of manufacture; being more preferred a pharmaceutical product comprising a vial containing a lyophilized composition consisting of 3 mg compound I, 10.2 mg phosphate calculated as potassium dihydrogen phosphate and 300 mg of sucrose. In some embodiments, the label affixed to the vial comprises an expiration date that is at least 24 months, at least 30 months, at least 36 months, at least 42 months, or at least 48 months from the date of manufacture. In some embodiments, the vial has a size of 5 mL to 50 mL, preferably 10 mL to 30 mL, such as 5 mL, 10 mL, 15 mL, 20 mL, 25 ml, 30 mL, 35 mL, 40 mL, 45 mL or 50 mL. In a preferred embodiment, the vial is a 10-30 mL vial, being more preferred a 10 mL vial. A vial size of 10 ml is optimized to overcome limitations of larger vial sizes which lead to production capacity reduction due to reduced freeze dryer capacity and also adequate extractable volumes due to size. A vial size of 10 mL overcomes both of these limitations.

[0136] As described herein, the solid-state forms of compound I as disclosed in the present invention may be used in compositions of the present invention. The compositions may be prelyophilization compositions. The solid-state forms of compound I as disclosed in the present invention may be used to manufacture compositions of the present invention. The compound I may comprise form B. The amount of form B as disclosed herein may vary and can be mixed with an amorphous form. Form B as disclosed herein may be used in the manufacturing process to prepare lyophilized bulk product. In some embodiments, the crystalline mixture may comprise other crystalline compound I (e.g. non form B crystalline compound I).

[0137] In further embodiments, the present invention relates to a pharmaceutical composition comprising compound I manufactured using form B of compound I and a pharmaceutically acceptable carrier. The pharmaceutical composition may no longer contain any form B compound I, however the composition manufacturing process utilized at least some form B in one or more steps. In further embodiments, the present invention relates to form B of compound I of use in the manufacture of a pharmaceutical composition comprising compound I. In yet further embodiments, the present invention relates to the use of form B of compound I in the manufacture of a pharmaceutical composition comprising compound I. In yet further embodiments, the present invention relates to form B of compound I for use as a medicament. Again, form B may no longer be present in the final composition but may be utilized during manufacturing. In yet further embodiments, the present invention further provides a method of treating any mammal, notably a human, affected by cancer which comprises administering to the affected individual a therapeutically effective amount of form B or compound I or of a pharmaceutical compositioncomprising form B of compound I and a pharmaceutically acceptable carrier; or a pharmaceutical composition made from a process utilizing form B of compound I.

[0138] In a further embodiment, the present invention relates to compound I having residual solvents of not more than 1 % w / w, 0.5% w / w, 0.1 % w / w or substantially not detected. In a further embodiment, the present invention relates to compound I having a water content of above 1.6% w / w, or of 1.7-5% w / w. In a further embodiment, the present invention relates to compound I having a water content of not more than 5%, 4% or 3% w / w.

[0139] The present invention encompasses compound I comprising at least a detectible amount of form B, up to 1 % w / w form B, up to 5% w / w form B, up to 10% w / w form B, up to 20% w / w form B, up to 30% w / w form B, up to 40% w / w form B, up to 50% w / w form B, up to 60% w / w form B, up to 70% w / w form B, up to 80% w / w form B, up to 90% w / w form B, up to 95% w / w form B, up to 98% w / w form B, or be substantially pure form B. In an embodiment, crystalline compound I as described herein may comprise at least a detective amount of form B, up to 1 % w / w form B, up to 5% w / w form B, up to 10% w / w form B, up to 20% w / w form B, up to 30% w / w form B, up to 40% w / w form B, up to 50% w / w form B, up to 60% w / w form B, up to 70% w / w form B, up to 80% w / w form B, up to 90% w / w form B, up to 95% w / w form B, up to 98% w / w form B, or be substantially pure form B. w / w is intended to mean the amount of compound I which is in the form B state. As such, purely by way of example, 50% w / w means the compound I API comprises 50% by weight form B and 50% by weight another form, for example amorphous form A.

[0140] In an embodiment, the present invention relates to pharmaceutical compositions comprising form B of compound I and a pharmaceutically acceptable carrier or manufactured from compound I comprising form B. The compound I used in the compositions or using during the manufacture of the compositions may comprise compound I comprising at least a detectible amount of form B, up to 1 % w / w form B, up to 5% w / w form B, up to 10% w / w form B, up to 20% w / w form B, up to 30% w / w form B, up to 40% w / w form B, up to 50% w / w form B, up to 60% w / w form B, up to 70% w / w form B, up to 80% w / w form B, up to 90% w / w form B, up to 95% w / w form B, up to 98% w / w form B, or be substantially pure form B. Crystalline compound I as disclosed herein may in embodiments comprise at least a detectible amount of form B, up to 1 % w / w form B, up to 5% w / w form B, up to 10% w / w form B, up to 20% w / w form B, up to 30% w / w form B, up to 40% w / w form B, up to 50% w / w form B, up to 60% w / w form B, up to 70% w / w form B, up to 80% w / w form B, up to 90% w / w form B, up to 95% w / w form B, up to 98% w / w form B, or be substantially pure form B. In alternative embodiments, other non-form B crystalline compound I may form crystalline compound I at the same w / w amounts.

[0141] The crystalline compound I as disclosed herein may be used to form pharmaceutical compositions according to the present invention. Accordingly, in embodiments crystalline compound I is used in the manufacture of a bulk compound I solution which is thereafter lyophilized to form the lyophilized compound I formulation. The crystalline compound I maycomprise form B as disclosed herein. Thus, in embodiments where reference is made to crystalline compound I it is intended to mean at least some form B.

[0142] Although the crystalline compound I may not be present in the final dosage form (due to the dissolution and subsequent lyophilization steps), it nevertheless may affect the properties of the final dosage form. By way of example, using crystalline compound I can reduce and / or simplify the total impurities including degradation products. Characteristic related substances profiles may demonstrate the use of crystalline compound I during manufacture. According to an embodiment, the total amount of related substances in the final lyophilized product may be not more that (NMT) 5.0% w / w, 4.0% w / w, 3.0% w / w, 2.5% w / w, 2.4% w / w, 2.3% w / w, 2.2% w / w, 2.1 % w / w, or 2.0% w / w. In another embodiment, the total amount of related substances in the final lyophilized product may be not more than (NMT) 5.0% area, 4.0% area, 3.0% area, 2.5% area, 2.4% area, 2.3% area, 2.2% area, 2.1 % area, or 2.0% area. In a preferred embodiment, the total related substances are NMT than 1 .9% w / w or NMT 1 .9% area. According to a further embodiment, the final lyophilized product comprises NMT 1 % w / w or NMT 1 % area of any unspecified impurity.

[0143] In an embodiment, the crystalline compound I used in the manufacture of the compositions disclosed herein may have an assay (% w / w) not less than 90% and a total impurities level lower than 5.0% area or lower 5.0% w / w. Specified impurities and their limits may be impurity A (<1.0% area), impurity B (<1.0% area), impurity D (<1.0% area), impurity E (<1.0% area), Impurity F (<1.0% area). Any other individual non-specified impurity may have a limit of <0.5% area.

[0144] Further specified impurities and their limits may be impurity A (<1.0% w / w), impurity B (<1 .0% w / w), impurity D (<1 .0% w / w), impurity E (<1 .0% w / w), Impurity F (<1 .0% w / w). Any other individual non-specified impurity may have a limit of <0.5% w / w.

[0145] In a particular embodiment according to the present invention, the crystalline compound I used in the manufacture of the compositions disclosed herein have an assay (% w / w) not less than 90% and a total impurities level lower than 5.0% area or lower 5.0% w / w. Specified impurities and their limits may be impurity A (<0.9% area), impurity B (<1 .0% area), impurity D (<0.7% area), impurity E (<0.7% area), Impurity F (<0.6% area). Any other individual non-specified impurity may have a limit of <0.5% area.

[0146] Further specified impurities and their limits may be impurity A (<0.9% w / w), impurity B (<1 .0% w / w), impurity D (<0.7% w / w), impurity E (<0.7% w / w), Impurity F (<0.6% w / w). Any other individual non-specified impurity may have a limit of <0.5% w / w.

[0147] In particular embodiments according to the present invention, lyophilized composition according to the present (utilizing the solid state form of compound I disclosed herein) comprise less than about 0.4% of impurity H (w / w based on compound I) when the composition ispackaged, and wherein upon storage at about 5 °C for about 24, 30 or 36 months the composition comprises less than about 1 .0% of impurity H (w / w based on compound I).

[0148] In particular embodiments according to the present invention, lyophilized composition according to the present (utilizing the solid-state form of compound I disclosed herein) comprise less than about 0.4% area of impurity H when the composition is packaged, and wherein upon storage at about 5 °C for about 24, 30 or 36 months the composition comprises less than about 1.0% area of impurity H.EXAMPLESAbbreviations abs AbsoluteAPI Active Pharmaceutical IngredientDAD Diode array detectorNMT No more thanXRPD X-ray powder diffractogramTLC Thin layer chromatographyUV Ultraviolet v / v volume ratio w / w weight ratioWFI Water for injection

[0149] The X-ray powder diffractogram (XRPD) was obtained with a D5000 diffractometer (Siemens) in Bragg-Brentano geometry with a graphite secondary monochromator, equipped with a Cu-Kai - Cu-Ka2 radiation source. The pattern was recorded at a tube voltage of 40 kV, tube current of 30 mA, applying a stepsize of 0.02002-theta with 10 seconds per step in the angular range of 5.0° to 100° 2-theta. Optics (splits): divergence (1 mm), antidispersion (0.2 mm), monochromator (0.6 mm). Primary and secondary Soller (2.3 °). A typical precision of the 2-theta values is in the range of about ± 0.202-theta. Thus a diffraction peak that appears at 5.0° 2-theta can appear between 4.8 and 5.22-theta on most X-ray diffractometers under standard conditions.

[0150] Compound 1 was prepared as described in Example 20 of WO 01 / 87895 A1.

[0151] Compound 4 was prepared as described in Example 19 of WO 2018 / 197663 A1.COMPARATIVE EXAMPLE 1

[0152] To a solution of 1 (1 equiv.) in acetic acid (12.5 mL / mmol) at 23 °C was added (S)-2-amino-3- (benzofuran-3-yl)-propan-1-ol (4) (12 equiv.). The reaction mixture was stirred during 72 hours and then the acetic acid was evaporated. Saturated aqueous solution of NaHCOs was added and the mixture was extracted with CH2CI2 and the combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under vacuum. Flash chromatography gave compound 5 (yield 20%). Table 1 summarizes the structural elucidation of compound 5.Table 1 .1H and13C NMR data of compound 5 in CDCH.a6 = Chemicals shifts are reported in ppm using residual CHCH (7.26 for1H and 77.0 for13C) as internal reference,bm= multiplicity,CJ = coupling constants are reported in Hz.ESI-MS m / z: 795.2 (M)+.COMPARATIVE EXAMPLE 2A) Synthesis of compound 6

[0153] 1 6To a solution of compound 1 (20.43 g, 32.86 mmol) in CH3CN (2 L, 0.16 M) was added (S)-2- amino-3-(benzofuran-3-yl)-propan-1-ol (4) (15.75 g, 82.36 mmol, 2.5 eq.) and cyanuric chloride (TCT) (6.31 g, 30% w / w). The reaction mixture was stirred at 85 °C for 23 h, cooled to 23 °C, and treated with a saturated aqueous solution of NaHCOs until pH 8-9 (1650 mL). The reaction mixture was extracted with CH2CI2 several times and the combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under vacuum to give crude 6 (42.83 g, chromatographic purity 33.1%). Flash chromatography with a gradient of Hex / EtOAc from 9:1 to 1 :9 followed by flash chromatography with Hex:EtOAc / CH2Cl2 / MeOH 6:2:2:0.1 gave compound 6 (12.47 g, 48% yield, chromatographic purity 97.7%, Batch A).

[0154] Table 2 summarizes the chromatographic impurity profile of crude 6 obtained in this example. Peaks with % area < 0.4 are not listed. The complete chromatographic profile is showed in Figure 1 .Table 2

[0155] Chromatographic analysis of crude 6 allowed the identification of compounds II and III as byproducts of the reaction.

[0156] Compound II was isolated by chromatography and characterized by NMR and mass spectrometry.Comp II1H NMR (500 MHz, CD3OD): 6 7.79 (dd, J = 12.8, 7.6 Hz, 1 H), 7.65 (d, J = 7.5 Hz, 1 H), 7.54 (m, 4H), 7.40 (dd, J = 8.2, 4.9 Hz, 2H), 7.21 (ddd, J = 8.4, 7.1 , 1 .3 Hz, 2H), 7.08 (t, J = 7.5 Hz, 2H), 4.37 (m, 1 H), 4.24 (dq, J = 7.8, 5.2 Hz, 1 H), 3.58 (m, 3H), 3.48 (dd, J = 11.0, 5.0 Hz, 1 H), 3.00(m, 2H), 2.84 (m, 2H).13C NMR (126 MHz, CD3OD): 5 169.3, 166.7, 156.7, 143.9, 143.8, 129.5, 125.2, 123.4, 123.4, 121.2, 120.7, 118.2, 118.2, 112.2, 1 12.1 , 64.0, 63.9, 62.5, 61.5, 53.5, 53.4, 26.1.ESI-MS m / z: 494.3 (M+H)+.

[0157] The structure of compound III was deduced from HPLC-MS data where the corresponding peak had a mass of 339.0 (M+H)+and its isotopic distribution showed the presence of two chlorine atoms in its structure.Comp III

[0158] The formation of compounds II and III significantly affect the purification and the yield of compound 6 when the reaction is scaled up because two chromatographic purifications were required to obtain it. Compounds II and III are removed with the first chromatography and other impurities of compound 6 are significantly reduced with the second chromatography.

[0159] Table 3 summarizes the chromatographic impurity profile of chromatographed 6. Peaks with %area < 0.4 are not listed. The complete chromatographic profile is shown in Figure 3.Table 3

[0160] Table 4 shows the yields of two additional batches obtained with this process.Table 4B) Synthesis of compound I.

[0161] 6To a solution of compound 6 (12.44 g, 15.65 mmol) in acetonitrile (620 mL) was added water (440 mL) and AgNOs (79.35 g, 467.1 mmol, 29.85 equiv.). The reaction mixture was stirred at 23 °C for 18 h in darkness and, after checking reaction completion by TLC, treated with a 1 :1 mixture of saturated aqueous solutions of NaHCOs and NaCI (1.38 L) for 15 min. The reaction mixture wasdiluted with CH2CI2 (1 .38 L) and stirred for 5 min before filtering over Celite 545 (540 g). The filtrate was decanted and the salts retained over the Celite were washed sequentially with CH2CI2 (2 x 1 .38 L) and EtOAc (3 x 1 .38 L). The filtrate was extracted with EtOAc (4 x 0.7 L). Combined organic layers were dried over Na2SO4 and evaporated to dryness under vacuum to give crude compound I (9.13 g, chromatographic purity 85.1 %). Crude I was purified by flash chromatography using a gradient of CH2Cl2:MeOH from 99:1 to 94:6). Selected fractions were combined and evaporated to dryness to give compound I (7.21 g, 59% yield, Batch AA, chromatographic purity 95.45%).

[0162] Table 5 shows the details of two additional batches obtained with this process Table 5COMPARATIVE EXAMPLE 3

[0163] Compound I (1.45 g, 18.45 mmol, chromatographic purity 97.6%) was dissolved in aqueous HCI (0.5 M, 175 mL) to give a cloudy solution that was treated with an aqueous solution of NH4CI / NH4OH (pH 9, 120 mL) to precipitate compound I, that was filtered and dried under vacuum to give 1 .33 g, 16.9 mmol, yield 91 .6%, chromatographic purity 96.8%.

[0164] Table 6 shows the impurity profile of compound I before and after precipitation.Table 6means not detected or lower than 0.05

[0165] The crystallized product has a lower purity than the starting material. Therefore, no improvement is achieved by this process.

[0166] Attempts of washing other acidic solutions of compound I with organic solvents such as dichloromethane resulted in a very significant transfer of compound I into the organic phase resulting in low recovery rates and lower purity of precipitated compound I.

[0167] From this data, it can be concluded that the acid-base precipitation that allows the purification of lurbinectedin (as described in WO2021 / 099635) does not provide a compound I form with a higher purity or a simplified impurity profile.EXAMPLE 1A) Synthesis of compound 6

[0168] 1 6

[0169] To a solution of compound 1 (20.08 g, 32.30 mmol) in CH3CN (325 mL, 16 mL / g) was added (S)-2-amino-3-(benzofuran-3-yl)-propan-1-ol (4) (12.35 g, 64.60 mmol, 2 equiv.) and trifluoroacetic acid (7.4 mL, 96.90 mmol, 3 eq.). The reaction mixture was stirred at 85 °C for 16.5 h and, after checking reaction completion by TLC, cooled to 23 °C, and treated with NH4OH / NH4CI buffer to pH 9 (500 mL). The reaction mixture was extracted with CH2CI2 three times and the combined organic layers were dried over Na2SO4, filtered and concentrated under vacuum to give crude 6 (33.40 g, chromatographic purity 63.5%). Flash chromatography(Hex:EtOAc / CH2Cl2 / MeOH 6:2:2:0.1) gives compound 6 (17.91 g, 70% yield, chromatographic purity 97.4%, Batch D).

[0170] Table 7 summarizes the chromatographic impurity profile of crude 6 obtained in this example. Peaks with % area < 0.4 are not listed. The complete chromatographic profile is showed in Figure 2.Table 7

[0171] Chromatographic analysis of crude and / or purified 6 allowed the identification of compounds 7, 8 and 9 as byproducts of this reaction.

[0172] Compound 7 was isolated by chromatography and characterized by NMR and mass spectrometry.1H NMR (400 MHz, CD3OD) 59.09 (brd, J= 8.5 Hz, 2H), 7.66 (m, 1H), 7.56 (d, J= 1.1 Hz, 1H),7.44 (dt, J= 8.4, 0.8 Hz, 1H), 7.26 (m, 2H), 4.31 (tq, J= 8.3, 6.0 Hz, 1H), 3.67 (m, 2H), 3.06 (ddd, J= 14.8, 6.0, 1.0 Hz, 1H),2.89 (ddd, J= 14.8, 8.3, 1.0 Hz, 1H).13C NMR (101 MHz, CD3OD) 6156.8, 143.8, 129.2, 125.4, 123.6, 120.6, 117.6, 112.2, 63.7, 53.6,53.5, 25.5.ES-API-MS m / z: 310.0 (M+Na)+, 288.0 (M+H)+.

[0173] Compounds 8 and 9 of formula:8 and 9were detected by HPLC-MS analysis of the reaction mixtures. The amount of compound 9 formed during the Pictet-Spengler reaction was significantly reduced by basic hydrolysis with NH4OH / NH4CI buffer at pH 9. Impurity levels of compounds 7 and 8 were not affected by this treatment.

[0174] Table 8 summarizes the profile of 6 after chromatographic purification. Peaks with % area < 0.4 are not listed. The complete profile is showed in Figure 4.Table 8

[0175] These results are substantially similar to those obtained in the manufacture of another three batches of compound 6 as showed in Table 9.Table 9B) Synthesis of compound I.

[0176] To a solution of compound 6 (17.9 g, 22.52 mmol, Batch D) in acetonitrile (135 mL, 7.35 mL / g) was added water (95 mL, 5.25 mL / g) and AgNOs (38.25 g, 225.19 mmol, 10 equiv.). The reaction mixture was stirred at 23 °C for 2.5 h in darkness and, after checking reaction completion by TLC, treated with a 1 :1 mixture of saturated aqueous solutions of NaHCOs and NaCI (1 .8 L) for 15 min. The reaction mixture was diluted with CH2CI2 (1 .8 L) and stirred for 5 min before filtering over Celite® 545. The filtrate was decanted and the salts retained over the Celite® were washed with mixtures of CH2Cl2:EtOAc 2:1 that were used afterwards to extract the aqueous phase of the filtrate. Combined organic layers were dried over anhydrous Na2SO4, filtered and evaporated to dryness under vacuum to give crude I (16.35 g, chromatographic purity 90.7%). Crude I waspurified by flash chromatography using a gradient of CH2Cl2:MeOH from 99:1 to 94:6) to afford 13.47 g of I, 76% yield with a chromatographic purity of 98.1% (Batch DD).

[0177] These results were substantially maintained in another batch obtained following the same process, as summarized in Table 10. Table 10

[0178] Table 11 compares the yields of Form A of compound I obtained by the process of Comparative Example 2 and the process of Example 1Table 11

[0179] It is remarkable that the improvements in the two synthetic steps described in Example 1 result in almost doubling the yields obtained in Comparative Example 2.

[0180] Table 12 compares the chromatographic profiles of Form A of compound I obtained as described in Comparative Example 2 and in Example 1.Table 12Not detected or lower than 0.05

[0181] Form A of compound I has a better chromatographic purity when obtained by the process of the present invention.Example 2. Further purification of compound I and manufacture of Form B of compound I

[0182] The process described in this example can be applied to Compound I obtained by the processes described in Comparative Example 2 or in Example 1 . a) Silver removal a1) with FeCIs

[0183] Compound I from Comparative Example 3 (50 mg, chromatographic purity 96.8%, silver content 1400 ppm) was dissolved in dichloromethane (20 mL) and the resulting solution waswashed with aqueous saturated FeCh (4 x 20 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated under vacuum to give 20 mg of compound I with a 91.3% chromatographic purity and a silver content of 19 ppm.

[0184] Although the use of FeCh resulted in remarkable reduction of silver content, it is accompanied by a poor recovery and a diminished chromatographic purity, 91.3% vs 96.8% in the starting material. This may be due to oxidation of compound I in presence of FeCh. a2) with Nal

[0185] Compound I from Comparative Example 3 (50 mg, chromatographic purity 96.8%, silver content 1400 ppm) was dissolved in dichloromethane (20 mL) and the resulting solution was washed with aqueous saturated Nal (4 x 20 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated under vacuum to give 49 mg of compound I with 97.5% chromatographic purity and a silver content of 5 ppm.

[0186] The use of Nal resulted in a very significant reduction of silver content with very good recovery accompanied by a slight increase of the chromatographic purity of compound I. a3) with Nal 20% w / w

[0187] Compound I from Example 1 B (13.47 g, Batch DD) was dissolved in dichloromethane (1340 mL) and the resulting solution was washed with 20% aqueous Nal (4 x 1340 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated under vacuum. b) Crystallization

[0188] The solid obtained in step a3) was dissolved in hot CH3CN (at 82 °C) at a concentration of 0.022 g / mL. The resulting solution was allowed to cool to room temperature (T < 30 °C) and treated with water until a permanent turbidity is visually detected (typically after addition of c.a. 30 mL of water per gram of I). The mixture was cooled to a temperature between 0 °C and 5 °C under magnetic agitation for 22 h. The mixture was filtered through a 0.45 pm PVDF membrane under vacuum and the resulting solid was dried under vacuum (p < 5 mbar) at 27 °C for 48 h to give 10.39 g of compound I with a chromatographic purity of 98.5% and 77% purification yield (Batch DD’).

[0189] Several batches of form A of compound I were processed using the method described in this example to give new batches of form B of compound I.

[0190] Table 13 compares the chromatographic profiles (% of area) of several batches of Forms A (AA, BB and CC) and B (AA’, BB’ and CC’) of compound I obtained by the process of comparative Example 2 and processed according to this example.Table 13‘Impurities at RRT 0.91 have different mass spectra.- Not detected or lower than 0.05% area

[0191] Table 14 summarizes the chromatographic profiles (% of area) of two batches of FormsA (FF and GG) and B (FF’ and GG’) of compound I obtained by the process of Example 1 and processed according to this example.Table 14

[0192] Form B of Compound I consistently presents fewer impurities than form A. This also results in higher chromatographic purity of form B compared with form A.

[0193] To sum up, the processes of the present invention present the following independent advantages over the prior art:Significantly increases the yield of compound 6 when prepared from 20 g of compound 1 ; 65-75% in Example 1 (A) compared to 48-52% of Comparative Example 2 (A). This yield improvement is achieved even with a reduction of the number of equivalents of benzofuran 4 from 2.5 to 2.Avoids sub-products II and III, whose formation during the Pictet-Spengler reaction consumes a significant amount of benzofuran 4, a valuable chemical intermediate, and require an additional chromatographic purification to obtain pharmaceuticalgrade compound 6.Removes a substantial amount of residual silver impurities.Significantly improves the yield of form A of compound I from compound 1 (53-59% vs. 20-30% of the processes described in comparative Example 2) while the scale is increased. This yield improvement is accompanied by an improvement in the chromatographic purity.Provides pharmaceutical-grade compound I as form B.Solid State Characterization:

[0194] Form B of compound I was characterized by XRPD.

[0195] XRPD angles 2-theta and their relative intensities of form B of compound I are summarized in Table 15. Peaks with Relative Intensity < 20% are not listed.Table 15 XRPD angles 2-theta, relative intensity of one batch of form B of compound I.Example 3 Process for the manufacture of a pharmaceutical composition using form B of compound I as starting material.

[0196] Form B of compound I (9.5 g) obtained by crystallization of a batch of compound I prepared as described in Comparative Example 2 and purified according to Example 2 (a3) was dissolved in a phosphoric acid solution (0.1 N) at a concentration of 5.7 mg / g. Then, this solution was diluted with water for injection (WFI) (5 L) and to this solution was added potassium dihydrogen phosphate (32.3 g) and sucrose (950 g). If required, the resulting solution can be adjusted to pH 2.8 - 3.3 with phosphoric acid solution (0.1 N) or potassium hydroxide solution (0.1 N).

[0197] Then, the bulk solution was brought to final volume or weight (considering a density value of 1.04 g / mL), generating the final bulk solution (1 mg / mL compound I, 3.4 mg / mL phosphate, expressed as potassium dihydrogen phosphate, 100 mg / mL sucrose).

[0198] Then, the bulk solution was then filtered through two sterilizing PVDF filters (0.2 pm) and filled into 10 mL vials at 3 mL / vial.

[0199] The vials were lyophilised according to a cycle detailed in Table 16.Table 16NA: Not applicable

[0200] After lyophilization, vials were sealed with flip-off seals and stored at +5 °C.Example 4 Stability of pharmaceutical composition of compound I manufactured as described in Example 3.

[0201] The purpose of this study was to investigate the stability of compound I in a formulation manufactured as described in Example 3.

[0202] A batch of 241 vials of the formulation of compound I was manufactured according to example 3.

[0203] Stability testing was carried out at a temperature of 5 °C ± 3 °C, 25 °C ± 2 °C / 60% RH ± 5% RH and 40 °C ± 2 °C / 75% RH ± 5% RH.

[0204] Table 17 shows the evolution of compound I content and total related substances at 5 °C ± 3 °CTable 17

[0205] Table 18 shows the evolution of the related-substance profile at 5 °C ± 3 °C expressed as % area Table 18- not detected or lower than 0.05% area

[0206] Table 19 shows the related substance profile at 5 °C ± 3 °C on month 28 expressed as % w / w.Table 19- not detected or lower than 0.05% w / w

[0207] Table 20 shows the evolution of compound I content and total related substances at 25°C ± 2 °C / 60% RH ± 5% RH. Table 20

[0208] Table 21 shows the evolution of the related-substance profile at 25 °C ± 2 °C / 60% RH ±5% RH expressed as % area.Table 21- not detected or lower than 0.05% area

[0209] Table 22 shows the evolution of compound I content and total related substances at 40 °C ± 2 °C / 75% RH ± 5% RH expressed as % respect to labelTable 22

[0210] Table 23 shows the evolution of the related-substance profile at 40 °C ± 2 °C / 75% RH ± 5% RH expressed as % areaTable 23- not detected or lower than 0.05% area.Conclusions

[0211] Stability data shows that a lyophilised composition of compound I prepared as described in Example 3 is stable after 28 months of storage at 5 °C ± 3 °C, after 12 months at 25 °C ± 2 °C / 60% RH ± 5% RH and after 6 months at 40 °C ± 2 °C / 75% RH ± 5% RH.CLAUSES1. A synthetic process comprising reacting compound 1 with benzofuran 4 in presence of trifluoroacetic acid to give compound 6.2. The process according to clause 1 , wherein the reaction is carried out using a solvent selected from acetonitrile and toluene.3. The process according to clause 2, wherein the solvent is acetonitrile.4. The process according to any preceding clause, wherein from about 2 to about 5 equivalents of trifluoroacetic acid are employed.5. The process according to clause 4, wherein about 3 equivalents of trifluoroacetic acid are employed.6. The process according to any preceding clause, wherein the reaction temperature is between about 80 °C and about 90 °C.7. The process according to clause 6, wherein the reaction temperature is about 85 °C.8. The process according to any preceding clause, wherein the reaction time is from about8 to about 24 hours.9. The process according to clause 8, wherein the reaction time is about 16 hours.10. The process according to any preceding clause, wherein the number of equivalents of benzofuran 4 is between about 1 .5 and about 5.11 . The process according to clause 10, wherein the number of equivalents of benzofuran 4 is about 2.12. The process according to any preceding clause, wherein the reaction mixture is treated with NH4OH / NH4CI buffer until the pH of the mixture is about 9.13. The process according to any preceding clause, wherein crude compound 6 is purified by flash chromatography under silica using as eluent a mixture of hexane, ethyl acetate, dichloromethane, and methanol in a 6:2:2:0.1 v / v ratio.14. A synthetic process for the synthesis of compound I or a pharmaceutically acceptable salt thereof, the process comprising reacting compound 6 with water in presence of AgNOs.6 I wherein the number of equivalents of AgNOs is from about 6 to about 15 and compound 6 is dissolved in a mixture of acetonitrile / water at an initial concentration from about 40 g / L to about 120 g / L.15. The process according to clause 14, wherein compound 6 is obtained as described in any one of clauses 1 to 13.16. The process according to clause 14 or clause 15, wherein the number of equivalents of AgNOs is from about 8 to about 12.17. The process according to clause 16, wherein the number of equivalents of AgNOs is about 10.18. The process according to any one of clauses 14 to 17, wherein the v / v ratio of acetonitrile / water is from about 3:1 to about 1 :1 .19. The process according to clause 18, wherein the acetonitrile / water v / v ratio is from about 1.5:1 to about 1.3:1.20. The process according to clause 19, wherein the acetonitrile / water v / v ratio is about 1 .4:1 .21. The process according to any one of clauses 14 to 20, wherein the initial concentration of compound 6 is from about 60 g / L to about 100 g / L.22. The process according to clause 18, wherein the initial concentration of compound 6 is about 79 g / L.23. A process for reducing silver impurities of compound I, the process comprising: a) dissolving compound I in a pharmaceutically acceptable, water-immiscible, polar organic solvent or a mixture of pharmaceutically acceptable, water-immiscible, polar organic solvents;b) washing such solution with an aqueous solution of a bromide or iodide salt, or mixtures thereof. c) decanting the organic phase, drying and filtering it; and d) evaporating the solvent to dryness.24. The process according to clause 23 wherein compound I is obtained by the process described in any one of clauses 14 to 22.25. The process according to clause 23 or clause 24, wherein the pharmaceutically acceptable, water-immiscible, polar organic solvent is dichloromethane.26. The process according to any one of clauses 23 to 25, wherein the bromide or iodide salt is an iodide salt or a mixture of iodide salts.27. The process according to clause 26, wherein the iodide salt is selected from Nal and KI.28. The process according to clause 27, wherein the iodide salt is Nal.29. The process according to any one of clauses 23 to 27, wherein the concentration of bromide or iodide salt in the aqueous solution if from about 10% to about 30% w / w.30. The process according to clause 29, wherein the concentration of the bromide or iodide salt in the aqueous solution is about 20% w / w.31. The process according to any one of clauses 23 to 30, wherein the washing step is repeated from about 3 to about 8 times.32. The process according to clause 31 , wherein the washing step is repeated from about 4 to about 5 times.33. The process according to any one of clauses 23 to 32, wherein the washing step is carried out after the chromatographic purification of compound I.34. A method of crystallization of compound I using the addition of an antisolvent.35. The method according to clause 34, wherein compound I has been previously obtained as defined in any one of clauses 14 to 22.36. The method according to clause 34, wherein compound I has been previously purified as defined in any one of clauses 23 to 33.37. The method according to clause 34, wherein compound I has been previously obtained as defined in any one of clauses 14 to 22 and it has been purified as defined in any one of clauses 23 to 33.38. The method according to any one of clauses 34 to 37, comprising the following steps:a) dissolving I in CH3CN at a temperature from about 75 °C to about 85 °C wherein the concentration of I is from about 0.020 to about 0.025 g / mL; b) cooling the solution to a temperature of from about 20 °C to about 30 °C; c) adding water until a dispersion of particles that does not re-dissolve is observed; d) maintaining the mixture at a temperature range of from about 0 °C to about 5 °C for a period of from about 15 to about 30 hours; wherein steps a) to d) are carried out with stirring.39. The method according to clause 38, further comprising the steps of e) separating crystalline compound I; f) washing the crystalline material with water; and g) drying the crystalline compound I under vacuum at a temperature of from about 25 °C to about 30 °C.40. A crystalline form of compound I of formula (I):l characterized by an X-ray powder diffraction pattern using Cu-Kaiand Cu-Kce radiation comprising three or more peaks at 2-theta angles selected from 9.5 ± 0.2°, 12.8 ± 0.2°, 14.8 ± 0.2°, 15.4 ± 0.2°, 16.2 ± 0.2° and 22.0 ± 0.2°.41 . The crystalline form according to clause 40, wherein the X-ray powder diffraction pattern using Cu-Kai and Cu-Kce radiation comprises four or more peaks at 2-theta angles selected from 9.5 ± 0.2°, 12.8 ± 0.2°, 14.8 ± 0.2°, 15.4 ± 0.2°, 16.2 ± 0.2° and 22.0 ± 0.2°.42. The crystalline form according to clause 41 , wherein the X-ray powder diffraction pattern using Cu-Kai and Cu-Kce radiation comprises five or more peaks at 2-theta angles selected fromthe group consisting of 9.5 ± 0.2°, 12.8 ± 0.2°, 14.8 ± 0.2°, 15.4 ± 0.2°, 16.2 ± 0.2° and 22.0 ± 0.2°.43. The crystalline form according to clause 40, wherein the X-ray powder diffraction pattern using Cu-Kaiand Cu-Ka2 radiation comprises peaks at 2-theta angles of 9.5 ± 0.2°, 14.8 ± 0.2° and 15.4 ± 0.2°.44. The crystalline form according to clause 41 , wherein the X-ray powder diffraction pattern using Cu-Kai and Cu-Kce radiation comprises peaks at 2-theta angles of 9.5 ± 0.2°, 12.8 ± 0.2°,14.8 ± 0.2° and 15.4 ± 0.2°.45. The crystalline form according to clause 42, wherein the X-ray powder diffraction pattern using Cu-Kai and Cu-Kce radiation comprises peaks at 2-theta angles of 9.5 ± 0.2°, 12.8 ± 0.2°,14.8 ± 0.2°, 15.4 ± 0.2° and 16.2 ± 0.2°.46. The crystalline form according to clause 40, wherein the X-ray powder diffraction pattern using Cu-Kai and Cu-Kce radiation comprises peaks at 2-theta angles of 9.5 ± 0.2°, 12.8 ± 0.2°,14.8 ± 0.2°, 15.4 ± 0.2°, 16.2 ± 0.2° and 22.0 ± 0.2°.47. The crystalline form according to clause 46, wherein the X-ray powder diffraction pattern using Cu-Kai and Cu-Kce radiation further comprises peaks at 2-theta angles of 10.0 ± 0.2°, 19.2 ± 0.2°, 19.9 ± 0.2°, and 21 .2 ± 0.2°.48. The crystalline form according to clause 46, wherein the X-ray powder diffraction pattern using Cu-Kai and Cu-Kce radiation further comprises peaks at 2-theta angles of 7.0 ± 0.2° and 7.5 ± 0.2°.49. The crystalline form according to clause 40, wherein the X-ray powder diffraction pattern using Cu-Kai and Cu-Kce radiation exhibits an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in Figure 5a and / or Figure 5b.50. The crystalline form according to clause 40 prepared by a process comprising the steps of a) dissolving I in CH3CN at a temperature from about 75 °C to about 85 °C wherein the concentration of I is from about 0.020 to about 0.025 g / mL; b) cooling the solution to a temperature of from about 20 °C to about 30 °C; c) adding water until a dispersion of particles that does not re-dissolve is observed; d) maintaining the mixture at a temperature range of from about 0 °C to about 5 °C for a period of from about 15 to about 30 hours;wherein steps a) to d) are carried out with stirring.51 . Compound I comprising at least a detectible amount of the crystalline form defined in any one of clauses 39 to 49, up to 1 % w / w of such form, up to 5% w / w of such form, up to 10% w / w of such form, up to 50% w / w of such form, up to 90% w / w of such form, or be substantially pure the crystalline form defined in any one of clauses 40 to 50.52. A composition comprising the crystalline form of compound I according to any one of clauses 40 to 51 and amorphous compound I.53. A pharmaceutical composition comprising the crystalline form of compound I as defined in any one of clauses 40 to 51 , and a pharmaceutically acceptable carrier; and / or a pharmaceutical composition comprising compound I manufactured via the crystalline form of compound I as defined in any one of clauses 40 to 51 , and a pharmaceutically acceptable carrier.54. A process for the manufacture of pharmaceutical compositions comprising compound I, said process employing the crystalline form of compound I defined in any one of clauses 40 to 51 , including as starting material.55. The process according to clause 54, comprising preparing a bulk solution for lyophilizing by dissolving the crystalline form of compound I defined in any one of clauses 40 to 51 in an acidic medium, adding the other components of the bulking solution and, optionally, adjusting the pH of the final solution.56. The process according to clause 55, further comprising freeze-drying the bulk solution.57. A method of manufacturing a pharmaceutical composition, comprising using the crystalline form of compound I as defined in any one of clauses 40 to 51 , to manufacture a pharmaceutical composition comprising compound I.58. A method of treating an individual affected by cancer comprising administering to said affected individual a therapeutically effective amount of the crystalline form of compound I defined in any one of claims 40 to 51 or a therapeutically effective amount of compound I which has been manufactured via the crystalline form B of compound I defined in any one of claims 40 to 51 .59. Crystalline compound I.60. Crystalline compound I according to clause 59, wherein the crystalline compound I comprises the crystalline form as defined in any one of clauses 40 to 51 ; and / or comprising at least a detectible amount of crystalline compound I, up to 1 % w / w crystalline compound I, up to 5% w / w crystalline compound I, up to 10% w / w crystalline compound I, up to 20% w / w crystalline compound I, up to 30% w / w crystalline compound I, up to 40% w / w crystallinecompound I, up to 50% w / w crystalline compound I, up to 60% w / w crystalline compound I, up to 70% w / w crystalline compound I, up to 80% w / w crystalline compound I, up to 90% w / w crystalline compound I, up to 95% w / w crystalline compound I, up to 98% w / w crystalline compound I, or substantially pure crystalline compound I; and / or comprising at least a detectible amount of the crystalline form defined in any one of clause 40 to 51 , up to 1% w / w of such form, up to 5% w / w of such form, up to 10% w / w of such form, up to 20% w / w of such form, up to 30% w / w of such form, up to 40% w / w of such form, up to 50% w / w of such form, up to 60% w / w of such form, up to 70% w / w of such form, up to 80% w / w of such form, up to 90% w / w of such form, up to 95% w / w of such form, up to 98% w / w of such form, or substantially pure the crystalline form defined in any one of clauses 40 to 51 ; and / or comprising at least a detectible amount of amorphous compound I, up to 1% w / w amorphous compound I, up to 5% w / w amorphous compound I, up to 10% w / w amorphous compound I, up to 20% w / w amorphous compound I, up to 30% w / w amorphous compound I, up to 40% w / w amorphous compound I, up to 50% w / w amorphous compound I, up to 60% w / w amorphous compound I, up to 70% w / w amorphous compound I, up to 80% w / w amorphous compound I, up to 90% w / w amorphous compound I, up to 95% w / w amorphous compound I, or up to 98% w / w amorphous compound I.61. A pharmaceutical composition or a pharmaceutical intermediate comprising crystalline compound I according to any one of clauses 40 to 51 or made from a process including crystalline compound I according to any one of clauses 40 to 51 .62. The composition of clause 61 , wherein the composition has a total water content of not more than 3%; and / or total related substances of not more than 5.0% w / w, 4.0% w / w, 3.0% w / w, 2.5% w / w, 2.3% w / w, 2.2% w / w, 2.1 % w / w or 2.0% w / w; and / or not more than 0.8% w / w of impurity H; and / or any unspecified related substances (highest) not more than 1.0% w / w; and / or wherein the pharmaceutical composition is a lyophilized composition.63. The composition of clause 61 , wherein the composition has a total water content of not more than 3%; and / or total related substances of not more than 5.0% area, 4.0% area, 3.0% area, 2.5% area, 2.3% area, 2.2% area, 2.1 % area or 2.0% area; and / or not more than 0.8% area of impurity H; and / or any unspecified related substances (highest) not more than 1 .0% area; and / or wherein the pharmaceutical composition is a lyophilized composition.64. A process for the manufacture of a compound I composition, said process employing compound I as defined in any one of clauses 40 to 51 ; including as a starting material.65. The process according to clause 64, wherein the process comprises pre-dissolving compound I in an inorganic acid, including wherein the inorganic acid has a pH less than 4, less than 3.5, less than 3, less than 2 or around 1 ; and / orwherein the inorganic acid has a normality of around 0.05 N to 0.5 N, around 0.05 N to 0.3 N, around 0.05 N to 0.15 N, or 0.1 N; and / or wherein the pre-dissolution step is at least 20 minutes, at least 30 minutes, at least 60 minutes, between 20-90 minutes, between 20-40 minutes; and / or wherein the solution is diluted with water for injection (WFI) to form a target concentration; and / or wherein the inorganic acid is phosphoric acid; and / or wherein to this solution it is added a basic ingredient and a bulking agent (e.g. disaccharide including sucrose) to form a buffered solution, including wherein the buffer solution has a pH of about 5.0 or less, about 2.5 to about 5.0, or about 2.5 to about 3.5, or about 2.8 to about 3.3; and further wherein the buffer is derived from phosphoric acid and potassium dihydrogen phosphate; and / or wherein the resulting solution is further diluted with water for injection (WFI) to form a target concentration of compound I, wherein the target concentration of compound I is optionally 1 mg / mL.66. The process according to clause 65, wherein the final target composition comprises 1 mg / mL of compound I, in phosphate buffer pH 2.8-3.3 and 100 mg / mL sucrose; and / or wherein a bulk solution is formed which undergoes sterilization filtrating before filing into vials; and / or wherein the composition undergoes freeze-drying to form a lyophilized formulation.67. The process according to clause 66, wherein the lyophilized composition is labelled for use; and / or wherein the lyophilized composition is reconstituted for use; and / or wherein the composition is reconstituted with from about 5 to about 50 mL of 0.9% sodium chloride solution to yield a solution having a pH of 2.8 to 3.3 and a compound I concentration of from about 0.06 to about 1 .0 mg / mL; and / or wherein a volume of reconstituted solution calculated to adjust the patient dosage is diluted to form an infusion solution with 0.9% sodium chloride solution, further optionally wherein the reconstituted solution is diluted with at least 100 mL or at least 250 mL to prepare a compound I infusion solution.68. The process as defined in clause 67, wherein the composition is reconstituted with from about 10 to about 30 mL of 0.9% sodium chloride solution to yield a solution having a pH of 2.8 to 3.3 and a compound I concentration of from about 0.06 to about 0.5 mg / mL.69 A compound I infusion solution or a reconstituted solution of a lyophilized composition or a bulk composition made according to the process of clause 65.70. A method of treating an individual affected by cancer comprising administering to said affected individual a therapeutically effective amount of crystalline compound I as defined in any one of clauses 40 to 51 or administering to said affected individual a therapeutically effective amount of a compound I composition manufacture using crystalline compound I as defined in any one of clauses 40 to 51 .71. A lyophilized pharmaceutical composition comprising compound I, wherein the composition is prepared by a process comprising:(a) dissolving a crystalline form of compound I in an aqueous solution having a pH less than 3.5;(b) dissolving a basic ingredient and a bulking agent;(c) freeze-drying the bulk solution to provide the lyophilized pharmaceutical composition comprising compound I, and wherein the crystalline form of compound I of step (a) is as defined in any one of clauses 40 to 51.72. The lyophilized pharmaceutical composition according to clause 71 , wherein the process further comprises diluting the solution resulting from step (a) with water for injection.73. The lyophilized pharmaceutical composition according to clause 71 or 72, wherein the process further comprises sterilizing the bulk solution by sterilization filtration.74. The lyophilized pharmaceutical composition according to any one of clauses 71 to 73, wherein the pH of the bulk solution is adjusted to a pH of 2.8 to 3.3 prior to lyophilization.75. The lyophilized pharmaceutical composition according to any one of clauses 71 to 74, wherein the aqueous solution having a pH less than 3.5 is a phosphoric acid solution.76. The lyophilized pharmaceutical composition according to any one of clauses 71 to 75, wherein the basic ingredient is potassium dihydrogen phosphate.77. The lyophilized pharmaceutical composition according to any one of clauses 71 to 76, wherein the bulking agent is a disaccharide.78. The lyophilized pharmaceutical composition according to clause 77, wherein the bulking agent is sucrose.79. The lyophilized pharmaceutical composition according to any one of clauses 71 to 78, wherein the lyophilized pharmaceutical compositions comprises no more than 1.0% w / w of impurity H based on the total weight of compound I.80. The lyophilized pharmaceutical composition according to clause 79, wherein total related substances of the lyophilized pharmaceutical composition is not more than about 5.0% w / w.81 . The lyophilized pharmaceutical composition according to clause 80, wherein total related substances of the lyophilized pharmaceutical composition is not more than 3.0% w / w.82. The lyophilized pharmaceutical composition according to clause 81 , wherein total related substances of the lyophilized pharmaceutical composition is not more than 2.5% w / w.83. The lyophilized pharmaceutical composition according to clause 82, wherein total related substances of the lyophilized pharmaceutical composition is not more than 2.0% w / w.84. The lyophilized pharmaceutical composition according to any one of clauses 71 to 83, wherein the lyophilized pharmaceutical composition has total related substances of no more than 5.0% w / w and not more than 1 % w / w of any unspecified substance.85. The lyophilized pharmaceutical composition according to any one of clauses 71 to 78, wherein the lyophilized pharmaceutical compositions comprises no more than 1.0% area of impurity H.86. The lyophilized pharmaceutical composition according to clause 85, wherein total related substances of the lyophilized pharmaceutical composition is not more than about 5.0% area.87. The lyophilized pharmaceutical composition according to clause 86, wherein total related substances of the lyophilized pharmaceutical composition is not more than 3.0% area.88. The lyophilized pharmaceutical composition according to clause 87, wherein total related substances of the lyophilized pharmaceutical composition is not more than 2.5% area.89. The lyophilized pharmaceutical composition according to clause 88, wherein total related substances of the lyophilized pharmaceutical composition is not more than 2.0% area.90. The lyophilized pharmaceutical composition according to any one of clauses 71 to 78, wherein the lyophilized pharmaceutical composition has total related substances of no more than 5.0% area and not more than 1 % area of any unspecified substance.91. A lyophilized pharmaceutical composition comprising compound I, wherein the composition is prepared by a process comprising: a) dissolving I in CH3CN at a temperature from about 75 °C to about 85 °C wherein the concentration of I is from about 0.020 to about 0.025 g / mL; b) cooling the solution to a temperature of from about 20 °C to about 30 °C; c) adding water until a dispersion of particles that does not re-dissolve in observed; d) maintaining the mixture at a temperature range of from about 0 °C to about 5 °C for a period of from about 15 to about 30 hours to precipitate a crystalline form of compound I, wherein steps a) to d) are carried out with stirring; and e) solubilizing the crystalline form of compound I and freeze drying the solubilized compound I to convert the crystalline form of compound I to an amorphous form to produce the lyophilized pharmaceutical composition.92. The lyophilized pharmaceutical composition according to clause 91 , wherein step e) comprises(1) dissolving the crystalline form of compound I in an inorganic acid solution;(2) dissolving a basic ingredient and a bulking agent; and(3) freeze-drying the bulk solution to provide the lyophilized pharmaceutical composition comprising amorphous compound I.93. The lyophilized pharmaceutical composition according to clause 91 or clause 92, wherein the process further comprises diluting the solution resulting from step (1) with water for injection.94. The lyophilized pharmaceutical composition according to any one of clauses 91 to 93, wherein the process further comprises sterilizing the bulk solution by sterilization filtration.95. The lyophilized pharmaceutical composition according to any one of clauses 91 to 94, wherein the pH of the bulk solution is adjusted to a pH of 2.8 to 3.3 prior to lyophilization.96. The lyophilized pharmaceutical composition according to any one of clauses 91 to 95, wherein the inorganic acid is phosphoric acid.97. The lyophilized pharmaceutical composition according to any one of clauses 91 to 96, wherein the basic ingredient is potassium dihydrogen phosphate.98. The lyophilized pharmaceutical composition according to any one of clauses 91 to 97, wherein the bulking agent is a disaccharide.99. The lyophilized pharmaceutical composition according to clause 98, wherein the bulking agent is sucrose.100. The lyophilized pharmaceutical composition according to any one of clauses 91 to 99, further comprising adding water to the bulk solution to obtain 1 mg / mL of compound 1 in a phosphate buffer at pH of 2.8-3.3 with 100 mg / mL sucrose.101. The lyophilized pharmaceutical composition according to any one of clauses 91 to 100, which comprises no more than 1 .0% w / w of impurity H based on the total weight of compound I.102. The lyophilized pharmaceutical composition according to any one of clauses 91 to 101 , wherein total related substances of the lyophilized pharmaceutical composition is not more than about 5.0% w / w.103. The lyophilized pharmaceutical composition according to clause 102, wherein total related substances of the lyophilized pharmaceutical composition is not more than 3.0% w / w.104. The lyophilized pharmaceutical composition according to clause 103, wherein total related substances of the lyophilized pharmaceutical composition is not more than 2.5% w / w.105. The lyophilized pharmaceutical composition according to clause 104, wherein total related substances of the lyophilized pharmaceutical composition is not more than 2.0% w / w.106. The lyophilized pharmaceutical composition according to any one of clauses 91 to 105, wherein the lyophilized pharmaceutical composition has total related substances of no more than 5.0% w / w and not more than 1 % w / w of any unspecified substance.107. The lyophilized pharmaceutical composition according to any one of clauses 91 to 100, which comprises no more than 1 .0% area of impurity H.108. The lyophilized pharmaceutical composition according to any one of clauses 91 to 100 and 107, wherein total related substances of the lyophilized pharmaceutical composition is not more than about 5.0% area.109. The lyophilized pharmaceutical composition according to clause 108, wherein total related substances of the lyophilized pharmaceutical composition is not more than 3.0% area.110. The lyophilized pharmaceutical composition according to clause 109, wherein total related substances of the lyophilized pharmaceutical composition is not more than 2.5% area.111. The lyophilized pharmaceutical composition according to clause 110, wherein total related substances of the lyophilized pharmaceutical composition is not more than 2.0% area.112. The lyophilized pharmaceutical composition according to any one of clauses 107 to 111 , wherein the lyophilized pharmaceutical composition has total related substances of no more than 5.0% area and not more than 1 % area of any unspecified substance.113. Compound I having a silver residual content of less than 1400 ppm.114. The compound according to clause 113 wherein the residual silver content is less than 1000 ppm.115. The compound according 'to clause 113 or clause 114, wherein the residual silver content is less than 500 ppm.116. The compound according to any one of clauses 113 to 115, wherein the residual silver content is less than 100 ppm.117. The compound according to any one of clauses 113 to 116 wherein the residual silver content is less than 10 ppm.

Claims

CLAIMSA synthetic process comprising reacting compound 1 with benzofuran 4 in presence of trifluoroacetic acid to give compound 6.

2. A synthetic process for the synthesis of compound I or a pharmaceutically acceptable salt thereof, the process comprising reacting compound 6 with water in presence of AgNOs.wherein the number of equivalents of AgNOs is from about 6 to about 15 and compound 6 is dissolved in a mixture of acetonitrile / water at an initial concentration from about 40 g / L to about 120 g / L.

3. The process according to claim 2, wherein compound 6 is obtained as described in claim 1.

4. A process for reducing silver impurities of compound I, the process comprising: a) dissolving compound I in a pharmaceutically acceptable, water-immiscible, polar organic solvent or a mixture of pharmaceutically acceptable, water-immiscible, polar organic solvents; b) washing such solution with an aqueous solution of a bromide or iodide salt, or mixtures thereof. c) decanting the organic phase, drying and filtering it; andd) evaporating the solvent to dryness.

5. The process according to claim 4, wherein compound I has been obtained as defined in claim 2 or 3.

6. A method of crystallization of compound I using the addition of an antisolvent.

7. The method according to claim 6, wherein the compound I used in the crystallization has been purified as defined in claim 4 or 5.

8. The method according to claim 6 or 7, comprising the following steps: a) dissolving I in CH3CN at a temperature from about 75 °C to about 85 °C wherein the concentration of I is from about 0.020 to about 0.025 g / mL; b) cooling the solution to a temperature of from about 20 °C to about 30 °C; c) adding water until a dispersion of particles that does not re-dissolve is observed; d) maintaining the mixture at a temperature range of from about 0 °C to about 5 °C for a period of from about 15 to about 30 hours; wherein steps a) to d) are carried out with stirring.

9. A crystalline form of compound I of formula (I):characterized by an X-ray powder diffraction pattern using Cu-Kaiand Cu-Kce radiation comprising three or more peaks at 2-theta angles selected from 9.5 ± 0.2°, 12.8 ± 0.2°, 14.8 ± 0.2°, 15.4 ± 0.2°, 16.2 ± 0.2° and 22.0 ± 0.2°.

10. A pharmaceutical composition comprising the crystalline form of compound I as defined in claim 9, and a pharmaceutically acceptable carrier; and / or a pharmaceutical composition comprising compound I manufactured via the crystalline form of compound I as defined in claim 9, and a pharmaceutically acceptable carrier.

11. A process for the manufacture of pharmaceutical compositions comprising compound I, said process employing the crystalline form of compound I defined in claim 9, including as starting material.

12. The process according to claim 11 , comprising preparing a bulk solution for lyophilizing by dissolving the crystalline form of compound I defined in claim 9 in an acidic medium, adding the other components of the bulking solution and, optionally, adjusting the pH of the final solution.

13. The process according to claim 12, comprising preparing a bulk solution for lyophilizing by dissolving the crystalline form of compound I defined in claim 9 in an acidic medium, adding the other components of the bulking solution and, optionally, adjusting the pH of the final solution.

14. The process according to claim 13, further comprising freeze-drying the bulk solution.

15. A lyophilized pharmaceutical composition comprising compound I, wherein the composition is prepared by a process comprising:(a) dissolving a crystalline form of compound I in an aqueous solution having a pH less than 3.5;(b) dissolving a basic ingredient and a bulking agent;(c) freeze-drying the bulk solution to provide the lyophilized pharmaceutical composition comprising compound I, and wherein the crystalline form of compound I of step (a) exhibits an X-ray power diffraction pattern using CuKai and CuKce radiation comprising three or more peaks at 2-theta angles selected from 9.5 ± 0.2°, 12.8 ± 0.2°, 14.8 ± 0.2°, 15.4 ± 0.2°, 16.2 ± 0.2° and 22.0 ± 0.2°.

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