Patent Publication Number: US-2006014734-A1

Title: Immediate release pharmaceutical formulation

Description:
This application is a national stage filing under 35 U.S.C. 371 of International Application No. PCT/SE03/00857, filed May 27, 2003, which claims priority from Sweeden Application No. 0201658-2, filed May 31, 2002, the specification of which is incorporated by reference herein. International Application No. PCT/SE03/00857 was published under PCT Article 21(2) in English. 
    
    
      This invention relates to a novel immediate release pharmaceutical formulation that provides for the delivery of particular pharmaceuticals, to the manufacture of such a formulation, and to the use of such a formulation in the treatment or prevention of thrombosis.  
      It is often desirable to formulate pharmaceutically active compounds for immediate release following oral and/or parenteral administration with a view to providing a sufficient concentration of drug in plasma within the time-frame required to give rise to a desired therapeutic response.  
      Immediate release may be particularly desirable in cases where, for example, a rapid therapeutic response is required (for example in the treatment of acute problems), or, in the case of parenteral administration, when peroral delivery to the gastrointestinal tract is incapable of providing sufficient systemic uptake within the required time-frame.  
      In the case of the treatment or prophylaxis of thrombosis, immediate release formulations may be necessary to ensure that a sufficient amount of drug is provided in plasma within a relatively short period of time to enable quick onset of action. Immediate release formulations are also typically simpler to develop than modified release formulations, and may also provide more flexibility in relation to the variation of doses that are to be administered to patients. Immediate release formulations are superior when multiple doses are not required and where it is not necessary to keep the plasma concentration at a constant level for an extended time.  
      International Patent Application No. PCT/SE01/02657 (WO 02/44145, earliest priority date 1 Dec. 2000, filed 30 Nov. 2001, published 6 Jun. 2002) discloses a number of compounds that are, or are metabolised to compounds which are, competitive inhibitors of trypsin-like proteases, such as thrombin. The following three compounds are amongst those that are specifically disclosed: 
      (a) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(OMe):  
                 
 
 which compound is referred to hereinafter as Compound A; 
    (b) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF)(OMe):  
                 
 
 which compound is referred to hereinafter as Compound B; and 
    (c) Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)—(S)Aze-Pab(OMe):  
                 
 
 which compound is referred to hereinafter as Compound C. 
   

      The methoxyamidine Compounds A, B and C are metabolised following oral and/or parenteral administration to the corresponding free amidine compounds, which latter compounds have been found to be potent inhibitors of thrombin. Thus: 
          Compound A is metabolized to Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab (which compound is referred to hereinafter as Compound D) via a prodrug intermediate Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(OH) (which compound is referred to hereinafter as Compound G);     Compound B is metabolized to Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF) (which compound is referred to hereinafter as Compound E) via a prodrug intermediate Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF)(OH) (which compound is referred to hereinafter as Compound H); and,     Compound C is metabolized to Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)—(S)Aze-Pab (which compound is referred to hereinafter as Compound F) via a prodrug intermediate Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)—(S)Aze-Pab(OH) (which compound is referred to hereinafter as Compound J).        

      Processes for the synthesis of Compounds A, B, C, D, E, F, G and J are described in Examples 12, 40, 22, 3, 39, 21, 2 and 31 (respectively) of international patent application No. PCT/SE01/02657. An immediate release formulation of these compounds, or their metabolites has yet to be described in the literature. We have found that the compounds of formula (I) and their salts can be formulated as immediate release pharmaceutical formulations which are easy to administer, for example by oral or parenteral administration.  
      According to a first aspect of the invention, there is provided an immediate release pharmaceutical formulation comprising, as active ingredient, a compound of formula (I):  
                 
 
 wherein 
      R 1  represents C 1-2  alkyl substituted by one or more fluoro substituents;     R 2  represents hydrogen, hydroxy, methoxy or ethoxy; and     n represents 0, 1 or 2; 
 
 or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable diluent or carrier; provided that the formulation does not solely contain: 
        a solution of one active ingredient and water;     a solution of one active ingredient and dimethylsulphoxide; or,     a solution of one active ingredient in a mixture of ethanol:PEG 660 12-hydroxy stearate:water 5:5:90; 
 
 which formulations are referred to hereinafter as “the formulations of the invention”. 
   
       

      PEG 660 12-hydroxy stearate is a non-ionic surfactant and is better known as Solutol K™.  
      According to a second aspect of the present invention there is provided Compound H, Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF)(OH) which can be prepared by methods similar to those described below for the preparation of Compounds G and J.  
      The compounds of formula (I), or a pharmaceutically acceptable salt thereof, may be in the form of a solvate, a hydrate, a mixed solvate/hydrate or, preferably, an ansolvate, such as an anhydrate. Solvates may be of one or more organic solvents, such as lower (for example C 1-4 ) alkyl alcohols (for example methanol, ethanol or iso-propanol), ketones (such as acetone), esters (such as ethyl acetate) or mixtures thereof.  
      In one particular aspect of the invention R 1  is CHF 2  or CH 2 CH 2 F.  
      The variable n is preferably 0 or 2.  
      More preferred compounds of formula (I) include those in which n represents 0, or those in which n represents 2, so providing two fluoro atoms located at the 2- and 6-positions (that is the two ortho-positions relative to the point of attachment of the benzene ring to the —NH—CH 2 — group).  
      The compound of formula (I) is especially Compound A, Compound B or Compound C.  
      Preferred salts of the compounds of formula (I) are acid addition salts. Acid addition salts include inorganic acid addition salts, such as those of sulphuric acid, nitric acid, phosphoric acid and hydrohalic acids, such as hydrobromic acid and hydrochloric acid. More preferred acid addition salts include those of organic acids, such as those of dimethylphosphoric acid; saccharinic acid; cyclohexylsulfamic acid; those of carboxylic acids (such as maleic acid, fumaric acid, aspartic acid, succinic acid, malonic acid, acetic acid, benzoic acid, terephthalic acid, hippuric acid, 1-hydroxy-2-naphthoic acid, pamoic acid, hydroxybenzoic acid and the like); those of hydroxy acids (such as salicylic acid, tartaric acid, citric acid, malic acid (including L-(−)-malic acid and, D,L-malic acid), gluconic acid (including D-gluconic acid), glycolic acid, ascorbic acid, lactic acid and the like); those of amino acids (such as glutamic acid (including D-glutamic, L-glutamic, and D,L-glutamic, acids), arginine (including L-arginine), lysine (including L-lysine and L-lysine hydrochloride), glycine and the like); and, particularly, those of sulfonic acids, (such as 1,2-ethanedisulfonic acid, camphorsulfonic acids (including 1S-(+)-10-camphorsulfonic acid and (+/−)-camphorsulfonic acids), ethanesulfonic acid, a propanesulfonic acid (including n-propanesulfonic acid), a butanesulfonic acid, a pentanesulfonic acid, a toluenesulfonic acid, methanesulfonic acid, p-xylenesulfonic acid, 2-mesitylenesulfonic acid, naphthalenesulfonic acids (including 1,5-naphthalenesulfonic acid and naphthalenesulfonic acid), benzenesulfonic acid, hydroxybenzenesulfonic acids, 2-hydroxyethanesulfonic acid, 3-hydroxyethanesulfonic acid and the like).  
      Particularly preferred salts include those of C 1-6  (for example C 1-4 ) alkanesulfonic acids, such as ethanesulfonic acid (esylate) and propanesulfonic acid (for example n-propanesulfonic acid) and optionally substituted (for example with one or more C 1-2  alkyl groups) arylsulfonic acids, such as benzenesulfonic acid (besylate) and naphthalenedisulfonic acid.  
      Suitable stoichiometric ratios of acid to free base are in the range 0.25:1.5 to 3.0:1, such as 0.45:1.25 to 1.25:1, including 0.50:1 to 1:1.  
      According to a further aspect of the invention there is provided formulation comprising a compound of formula (I) in substantially crystalline form.  
      Although we have found that it is possible to produce compounds of the invention in forms which are greater than 80% crystalline, by “substantially crystalline” we include greater than 20%, preferably greater than 30%, and more preferably greater than 40% (e.g. greater than any of 50, 60, 70, 80 or 90%) crystalline.  
      According to a further aspect of the invention there is also provided a compound of the invention in partially crystalline form. By “partially crystalline” we include 5% or between 5% and 20% crystalline.  
      The degree (%) of crystallinity may be determined by the skilled person using X-ray powder diffraction (XRPD). Other techniques, such as solid state NMR, FT-IR, Raman spectroscopy, differential scanning calorimetry (DSC) and microcalorimetry, may also be used.  
      Preferred compounds of formula (I) that may be prepared in crystalline form include salts of C 1-6  (for example C 2-6 , such as C 2-4 ) alkanesulfonic acids, such as ethanesulfonic acid, propanesulfonic acid (for example n-propanesufonic acid) and optionally substituted arylsulfonic acids, such as benzenesulfonic acid and naphthalenedisulfonic acid.  
      The term “immediate release” pharmaceutical formulation includes any formulation in which the rate of release of drug from the formulation and/or the absorption of drug, is neither appreciably, nor intentionally, retarded by galenic manipulations. In the present case, immediate release may be provided for by way of an appropriate pharmaceutically acceptable diluent or carrier, which diluent or carrier does not prolong, to an appreciable extent, the rate of drug release and/or absorption. Thus, the term excludes formulations which are adapted to provide for “modified”, “controlled”, “sustained”, “prolonged”, “extended” or “delayed” release of drug.  
      In this context, the term “release” includes the provision (or presentation) of drug from the formulation to the gastrointestinal tract, to body tissues and/or into systemic circulation. For gastrointestinal tract release, the release is under pH conditions such as pH=1 to 3, especially at, or about, pH=1. In one aspect of the invention a formulation as described herein with a compound of formula (I), or an acid addition salt thereof, in crystalline form releases drug under a range of pH conditions. In another aspect of the invention a formulation as described herein with a compound of formula (I), or an acid addition salt thereof, releases drug under pH conditions such as pH=1 to 3, especially at, or about, pH=1. Thus, formulations of the invention may release at least 70% (preferably 80%) of active ingredient within 4 hours, such as within 3 hours, preferably 2 hours, more preferably within 1.5 hours, and especially within an hour (such as within 30 minutes), of administration, whether this be oral or parenteral.  
      The formulations of the invention may be formulated in accordance with a variety of known techniques, for example as described by M. E. Aulton in “ Pharmaceutics: The Science of Dosage Form Design ” (1988) (Churchill Livingstone), the relevant disclosures in which document are hereby incorporated by reference.  
      Formulations of the invention may be, or may be adapted in accordance with standard techniques to be, suitable for peroral administration, for example in the form of an immediate release tablet, an immediate release capsule or as a liquid dosage form, comprising active ingredient. These formulation types are well known to the skilled person and may be prepared in accordance with techniques known in the art.  
      Suitable diluents/carriers (which may also be termed “fillers”) for use in peroral formulations of the invention, for example those in the form of immediate release tablets, include monobasic calcium phosphate, dibasic calcium phosphate (including dibasic calcium phosphate dihydrate and dibasic calcium phosphate anhydrate), tribasic calcium phosphate, lactose, microcrystalline cellulose, silicified microcrystalline cellulose, mannitol, sorbitol, starch (such as maize, potato or rice), glucose, calcium lactate, calcium carbonate and the like. Preferred diluents/carriers include dibasic calcium phosphate and microcrystalline cellulose, which may be used alone or in combination with another diluent/carrier such as mannitol.  
      A formulation of the invention in the form of an immediate release tablet may comprise one or more excipients to improve the physical and/or chemical properties of the final composition, and/or to facilitate the process of manufacture. Such excipients are conventional in the formulation of immediate release formulations for peroral drug delivery, and include one or more of the following: one or more lubricants (such as magnesium stearate, stearic acid, calcium stearate, stearyl alcohol or, preferably, sodium stearyl fumarate); a glidant (such as talc or a colloidal silica); one or more binders (such as polyvinylpyrrolidone, microcrystalline cellulose, a polyethylene glycol (PEG), a polyethylene oxide, a hydroxypropylmethylcellulose (HPMC) of a low molecular weight, a methylcellulose (MC) of a low molecular weight, a hydroxypropylcellulose (HPC) of a low molecular weight, a hydroxyethylcellulose (HEC) of a low molecular weight, a starch (such as maize, potato or rice) or a sodium carboxymethyl cellulose of a low molecular weight; (preferred binders are polyvinylpyrrolidone or a HPMC of a low molecular weight); one or more pH controlling agents (such as an organic acid (for example citric acid) or an alkali metal (for example sodium) salt thereof, an oxide of magnesium, an alkali or alkaline earth metal (for example sodium, calcium or potassium) sulphate, metabisulphate, propionate or sorbate); one or more disintegrant (for example sodium starch glycollate, a crosslinked polyvinylpyrrolidone, a crosslinked sodium carboxymethyl cellulose, a starch (such as maize, potato or rice) or an alginate); a colourant, a flavouring, a tonicity-modifying agent, a coating agent or a preservative.  
      It will be appreciated that some of the above mentioned excipients which may be present in a final immediate release oral (for example tablet) formulation of the invention may have more than one of the above-stated functions.  
      In a further aspect of the invention a liquid formulation of the invention is adapted to be suitable for oral administration.  
      Suitable liquid formulations that are to be administered orally include those in which a compound of formula (I) especially Compound A, Compound B or Compound C, or a pharmaceutically acceptable salt thereof is presented together with an aqueous carrier, such as water. It will be noted however, that certain specific formulations are not claimed (see particular aspects and the claims).  
      A formulation of the present invention comprising an aqueous carrier may further comprise one or more excipients, such as an antimicrobial preservative; a tonicity modifier (for example sodium chloride, mannitol or glucose); a pH adjusting agent (for example a common inorganic acid or base, including hydrochloric acid or sodium hydroxide); a pH controlling agents (that is, a buffer; for example tartaric acid, acetic acid or citric acid); a surfactant (for example Sodiun dodecyl sulphate (SDS) or Solutol™); a solubiliser which serves to help solubilise the active ingredient (for example ethanol, a polyethylene glycol or hydroxypropyl-β-cyclodextrin or polyvinyl chloride (PVP)); or an antioxidant.  
      Liquid oral formulations may be in the form of suspensions of active ingredient in association with an aqueous solvent or, more preferably aqueous solutions (that is, solutions of active compound including water as a solvent). In this context, the term “aqueous solution” includes formulations in which at least 99% of active ingredient is in solution at above 5° C. and atmospheric pressure, and the term “suspension” means that more than 1% of active ingredient is not in solution under such conditions. Typical dispersion agents for suspensions are hydroxypropyl methylcellulose, AOT (dioctylsulfosuccinate), PVP and SDS. Other alternatives may be possible.  
      In another aspect the present invention provides a liquid oral formulation comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, water and at least one additional agent. The additional agents include: 
          i. polyethylene glycol (PEG) and optionally also ethanol and/or tartaric acid and/or citric acid and/or hydrochloric acid; or     ii. sodium chloride (which will be dissolved in the formulation), and optionally also ethanol; or     iii. hydrochloric acid and/or sodium hydroxide to bring the pH to a suitable value (preferably in the range 3-8 for a compound of formula (I) wherein R 2  is methoxy or ethoxy, such as Compound A, B or C); or     iv. DMA (dimethyl acetamide) and optionally also a medium chain triglyceride (such as miglyol); or     v. a β-cyclodextrin (such as hydroxypropyl-β-cyclodextrin);     vi. a tonicity modifier such as sodium chloride and/or mannitol.        

      In a further aspect the present invention provides an oral solution comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, (preferably Compound A, B or C) water and at least one additional agents as recited in (i) to (vi) above.  
      In another aspect the invention provides an aqueous formulation of a compound of formula (I) (such as Compound A, B or C) comprising a solubilising agent such as a polyethylene glycol, β-cyclodextrin (such as hydroxypropyl-β-cyclodextrin), sorbitol or ethanol.  
      In a further aspect the present invention provides an oral solution formulation comprising a compound of formula (I) and ethanol. This formulation can further comprise a medium chain triglyceride (such as miglyol).  
      In a still further aspect the present invention provides an oral solution formulation comprising a compound of formula (I) and DMA. This formulation can further comprise a medium chain triglyceride (such as miglyol).  
      In another aspect the compound of formula (I) is crystalline (especially a salt of Compound A; preferably a C 1-6  (for example C 2-6 , such as C 2-4 ) alkanesulfonic acid salt, such as ethanesulfonic acid, propanesulfonic acid (for example n-propanesufonic acid) or an optionally substituted arylsulfonic acid salt, such as benzenesulfonic acid or naphthalenedisulfonic acid salt).  
      A particular liquid immediate release oral pharmaceutical formulation as claimed in claim  1  is provided wherein the active ingredient is: 
      Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(OMe),     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF)(OMe),     Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)—(S)Aze-Pab(OMe), 
 
 or a pharmaceutically acceptable salt thereof. 
   

      A further particular liquid immediate release oral pharmaceutical formulation as claimed in claim  1  is provided wherein the active ingredient is: Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(OMe) or a C 1-4  alkanesulfonic acid or an optionally substituted arylsulfonic acid salt thereof  
      A yet further particular liquid immediate release oral pharmaceutical formulation as claimed in claim  1  is provided wherein the active ingredient is: Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF)(OMe) or an optionally substituted arylsulfonic acid salt thereof (such as the naphthalene-1,5-disulphonic acid salt).  
      It will be noted however, that certain specific formulations are not claimed (see particular aspects and the claims).  
      In a further aspect of the invention a formulation of the invention is adapted to be suitable for parenteral administration. The term “parenteral” includes any mode of administration that does not comprise peroral administration to the gastrointestinal tract and includes administration subcutaneously, intravenously, intraarterially, transdermally, intranasally, intrabuccally, intracutaneously, intramuscularly, intralipomateously, intraperitoneally, rectally, sublingually, topically, by inhalation, or by any other parenteral route.  
      Suitable formulations of the invention that are to be administered parenterally include those in which a compound of formula (I) or a pharmaceutically acceptable salt thereof is presented together with an aqueous carrier, such as water.  
      A formulation of the present invention comprising an aqueous carrier may further comprise one or more excipients, such as an antimicrobial preservative; a tonicity modifier (for example sodium chloride, mannitol or glucose); a pH adjusting agent (for example a common inorganic acid or base, including hydrochloric acid or sodium hydroxide); a pH controlling agents (that is, a buffer; for example tartaric acid, acetic acid or citric acid); a surfactant (for example sodium dodecyl sulphate (SDS) or Solutol™); a solubiliser which serves to help solubilise the active ingredient (for example ethanol, a polyethylene glycol or hydroxypropyl-β-cyclodextrin or polyvinyl chloride (PVP)); or an antioxidant.  
      Parenteral formulations may be in the form of suspensions of active ingredient in association with an aqueous solvent or, more preferably aqueous solutions (that is, solutions of active compound including water as a solvent). In this context, the term “aqueous solution” includes formulations in which at least 99% of active ingredient is in solution at above 5° C. and atmospheric pressure, and the term “suspension” means that more than 1% of active ingredient is not in solution under such conditions. Typical dispersion agents for suspensions are hydroxypropyl methylcellulose, AOT, PVP and SDS, but other alternatives are possible.  
      The number of excipients employed in the peroral and parenteral formulations of the invention depends upon many factors, such as the nature and amount of active ingredient present, and the amount of diluent/carrier (aqueous solvent or otherwise) that is included.  
      In another aspect the present invention provides a parenteral formulation comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, water and at least one additional agents. The additional agents include: 
          i. polyethylene glycol (PEG) and optionally also ethanol and/or tartaric acid and/or hydrochloric acid; or     ii. sodium chloride (which will be dissolved in the formulation), and optionally also ethanol; or     iii. hydrochloric acid and/or sodium hydroxide to bring the pH to a suitable value (preferably in the range 3-8 for a compound of formula (I) wherein R 2  is hydrogen, such as Compound D, E or F; or preferably in the range 3.5-8 for a compound of formula (I) wherein R 2  is methoxy or ethoxy, such as Compound A, B or C); or     iv. DMA (dimethyl acetamide) and optionally also a medium chain triglyceride (such as miglyol); or     v. a β-cyclodextrin (such as hydroxypropyl-β-cyclodextrin);     vi. a tonicity modifier such as sodium chloride and/or mannitol.        

      In a further aspect the present invention provides an injectable solution comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, (preferably Compound D, E or F) water and at least one additional agents as recited in (i) to (vi) above.  
      In another aspect the invention provides an aqueous formulation of a compound of formula (I) (such as Compound D, E or F) comprising a solubilising agent such as a polyethylene glycol, β-cyclodextrin (such as hydroxypropyl-β-cyclodextrin), sorbitol or ethanol.  
      In a further aspect the present invention provides a parenteral formulation comprising a compound of formula (I) and ethanol. This formulation can further comprise a medium chain triglyceride (such as miglyol).  
      In a still further aspect the present invention provides a parenteral formulation comprising a compound of formula (I) and DMA. This formulation can further comprise a medium chain triglyceride (such as miglyol).  
      In another aspect the compound of formula (I) is crystalline (especially a salt of Compound A; preferably a C 1-6  (for example C 2-6 , such as C 2-4 ) alkanesulfonic acid salt, such as ethanesulfonic acid, propanesulfonic acid (for example n-propanesufonic acid) or an optionally substituted arylsulfonic acid salt, such as benzenesulfonic acid salt).  
      In yet another aspect the formulation of the present invention is in a solid dosage form wherein R 2  is hydroxy, methoxy or ethoxy (preferably methoxy) (the compound of formula (I) is especially Compound A, Compound B or Compound C).  
      In yet another aspect the present invention provides a parenteral formulation (especially a water-based, injectable solution) comprising a compound of formula (I) in free base form.  
      In a further aspect the present invention provides a parenteral formulation comprising a compound of formula (I) in free base form wherein R 2  is hydrogen.  
      In a still further aspect the present invention provides a solid formulation comprising microcrystalline cellulose and polyvinyl pyrrolidone (PVP); or comprising microcrystalline cellulose and sodium starch glycollate.  
      Formulations of the invention, such as parenteral formulations, comprising salts may be prepared by addition of diluent/carrier to the appropriate pre-prepared salt.  
      Compositions including active ingredient may also be provided in solid form suitable for use in the preparation of a formulation of the invention (for example a solution, such as an aqueous solution, for example for parenteral adminstration) ex tempore. Such compositions may be in the form of a solid comprising active ingredient, optionally in the presence of one or more further excipients as hereinbefore defined and, optionally, up to 10% (w/w) of diluent and/or carrier as hereinbefore defined, which compositions are hereinafter referred to as “the solid compositions of the invention”.  
      Solid compositions of the invention may be made by removal of diluent/carrier (for example solvent) from a formulation of the invention, or a concentrated formulation of the invention, which may for example be in the form of a solution, such as an aqueous solution.  
      In another aspect the present invention provides an orally administerable, immediate release formulation comprising a compound of formula (I), or a salt thereof, a carrier (such as microcrystalline cellulose), a disintegrant (such as sodium starch glycollate), a binder (such as polyvinyl pyrrolidone) and a lubricant (such as sodium stearyl fumarate). Such a formulation may also comprise an additional carrier (or filler) such as mannitol.  
      Formulations of the invention that are in the form of immediate release tablets may be prepared by bringing active ingredient into association with diluent/carrier using standard techniques, and using standard equipment, known to the skilled person, including wet or dry granulation, direct compression/compaction, drying, milling, mixing, tableting and coating, as well as combinations of these processes, for example as described hereinafter. In one aspect of the invention, acid addition salts of compounds of formula (I) in crystalline form are formulated in tablets.  
      There is thus provided a process for the formation of a solid composition suitable for use in the preparation of a formulation of the invention (for example a solution, such as an aqueous solution) ex tempore, which process comprises removal of diluent/carrier (for example solvent) from a formulation of the invention, or a concentrated formulation of the invention.  
      Solvent may be removed by way of a variety of techniques known to those skilled in the art, for example evaporation (under reduced pressure or otherwise), freeze-drying, or any solvent removal (drying) process that removes solvent (such as water) while maintaining the integrity of the active ingredient. An example of drying is freeze-drying.  
      Thus according to a further aspect of the invention there is provided a freeze-dried (lyophilised) solid composition of the invention.  
      In the preparation of solid compositions of the invention, the skilled person will appreciate that appropriate additional excipients may be added at a suitable stage prior to removal of diluent/carrier. For example, in the case of aqueous solutions, pH may be controlled and/or adjusted as hereinbefore described. Furthermore, an appropriate additional excipient may be added with a view to aiding the formation of a solid composition of the invention during the process of diluent/carrier removal (for example mannitol, sucrose, glucose, mannose or trehalose).  
      A solid composition of a compound of formula (I) or a salt thereof, thus includes a composition in which the solvent (for example water) content, other than a solvent of crystallization, is no more than 10%, such as less than 2% unbound solvent, such as water.  
      Formulations of the invention may be sterilised, for example by sterile filtration or autoclavation, and/or filled into primary packages, such as vials, cartridges and pre-filled syringes. Such processing steps may also take place prior to drying to form a solid composition of the invention.  
      Before administration, the dried solid composition may be reconstituted and/or diluted in, for instance, water, physiological saline, glucose solution or any other suitable solution.  
      The amount of diluent/carrier in an oral (for example immediate release tablet) formulation of the invention depends upon many factors, such as the nature and amount of the active ingredient that is employed and the nature, and amounts, of any other constituents (for example further excipients) that are present in the formulation, but is typically up to 40% (w/w), preferably up to 30%, more preferably up to 20%, and particularly up to 10% (w/w) of the final composition. The amount of additional excipients in such an oral formulation of the invention also depends upon factors, such as the nature and amount of the active ingredient that is employed, as well as the nature, and amounts, of any other constituents (for example diluents/carriers and/or other further excipients) that are present in the formulation, but, for lubricants and glidants is typically up to 5% (w/w), and for binders and disintegrants is typically up to 10% (w/w) of the final composition.  
      The formulations of the invention are administered to mammalian patients (including humans), and, for compounds of formula (I) wherein R 2  is not hydrogen, are thereafter metabolised in the body to form compounds of formula (I) wherein R 2  is hydrogen that are pharmacologically active.  
      According to a further aspect of the invention there is thus provided a formulation of the invention for use as a pharmaceutical.  
      In particular, the compounds of formula (I) are, or are metabolised following administration to form, potent inhibitors of thrombin, for example as may be demonstrated in the tests described in inter alia international patent application No. PCT/SE01/02657, as well as international patent applications WO 02/14270, WO 01/87879 and WO 00/42059, the relevant disclosures in which documents are hereby incorporated by reference.  
      By “prodrug of a thrombin inhibitor”, we include compounds that are metabolised following administration and form a thrombin inhibitor, in an experimentally-detectable amount, following administration.  
      By “active ingredient” and “active substance” we mean the pharmaceutical agent (covering thrombin inhibitor and prodrugs thereof) present in the formulation.  
      The formulations of the invention are thus expected to be useful in those conditions where inhibition of thrombin is required, and/or conditions where anticoagulant therapy is indicated, including the following:  
      The treatment and/or prophylaxis of thrombosis and hypercoagulability in blood and/or tissues of animals including man. It is known that hypercoagulability may lead to thrombo-embolic diseases. Conditions associated with hypercoagulability and thrombo-embolic diseases which may be mentioned include inherited or acquired activated protein C resistance, such as the factor V-mutation (factor V Leiden), and inherited or acquired deficiencies in antithrombin III, protein C, protein S, heparin cofactor II. Other conditions known to be associated with hypercoagulability and thrombo-embolic disease include circulating antiphospholipid antibodies (Lupus anticoagulant), homocysteinemi, heparin induced thrombocytopenia and defects in fibrinolysis, as well as coagulation syndromes (for example disseminated intravascular coagulation (DIC)) and vascular injury in general (for example due to surgery).  
      The treatment of conditions where there is an undesirable excess of thrombin without signs of hypercoagulability, for example in neurodegenerative diseases such as Alzheimer&#39;s disease.  
      Particular disease states which may be mentioned include the therapeutic and/or prophylactic treatment of venous thrombosis (for example DVT) and pulmonary embolism, arterial thrombosis (e.g. in myocardial infarction, unstable angina, thrombosis-based stroke and peripheral arterial thrombosis), and systemic embolism usually from the atrium during atrial fibrillation (for example non-valvular atrial fibrillation) or from the left ventricle after transmural myocardial infarction, or caused by congestive heart failure; prophylaxis of re-occlusion (that is thrombosis) after thrombolysis, percutaneous trans-luminal angioplasty (PTA) and coronary bypass operations; the prevention of re-thrombosis after microsurgery and vascular surgery in general.  
      Further indications include the therapeutic and/or prophylactic treatment of disseminated intravascular coagulation caused by bacteria, multiple trauma, intoxication or any other mechanism; anticoagulant treatment when blood is in contact with foreign surfaces in the body such as vascular grafts, vascular stents, vascular catheters, mechanical and biological prosthetic valves or any other medical device; and anticoagulant treatment when blood is in contact with medical devices outside the body such as during cardiovascular surgery using a heart-lung machine or in haemodialysis; the therapeutic and/or prophylactic treatment of idiopathic and adult respiratory distress syndrome, pulmonary fibrosis following treatment with radiation or chemotherapy, septic shock, septicemia, inflammatory responses, which include, but are not limited to, edema, acute or chronic atherosclerosis such as coronary arterial disease and the formation of atherosclerotic plaques, cerebral arterial disease, cerebral infarction, cerebral thrombosis, cerebral embolism, peripheral arterial disease, ischaemia, angina (including unstable angina), reperfusion damage, restenosis after percutaneous trans-luminal angioplasty (PTA) and coronary artery bypass surgery.  
      The formulation of the present invention may also comprise any antithrombotic agent(s) with a different mechanism of action to that of the compounds of formula (I), such as one or more of the following: the antiplatelet agents acetylsalicylic acid, ticlopidine and clopidogrel; thromboxane receptor and/or synthetase inhibitors; fibrinogen receptor antagonists; prostacyclin mimetics; phosphodiesterase inhibitors; ADP-receptor (P 2 T) antagonists; and inhibitors of carboxypeptidase U (CPU).  
      Compounds of formula (I) that inhibit trypsin and/or thrombin may also be useful in the treatment of pancreatitis.  
      The formulations of the invention are thus indicated both in the therapeutic and/or prophylactic treatment of these conditions.  
      According to a further aspect of the present invention, there is provided a method of treatment of a condition where inhibition of thrombin is required which method comprises administration of a therapeutically effective amount of a formulation of the invention to a person suffering from, or susceptible to, such a condition.  
      In a still further aspect the present invention provides a formulation of the invention in the manufacture of a medicament for use in the treatment of thrombosis.  
      According to a further aspect of the invention, there is provided a method of treatment of thrombosis which method comprises administration of a formulation of the invention to a person suffering from, or susceptible to, such a condition.  
      For the avoidance of doubt, by “treatment” we include the therapeutic treatment, as well as the prophylaxis, of a condition.  
      Suitable amounts of active ingredient in formulations (oral or parenteral), concentrated formulations, and solid compositions, of the invention depend upon many factors, such as the nature of that ingredient (free base/salt etc), the dose that is required in an oral formulation or in a final “ready to use” parenteral formulation that is, or is to be, prepared, and the nature, and amounts, of other constituents of the formulation. However, a typical daily dose of a compound of formula (I), or a pharmaceutically acceptable salt thereof, is in the range 0.001-100 mg/kg body weight at peroral administration and 0.001-50 mg/kg body weight at parenteral administration, excluding the weight of any acid counter-ion, irrespective of the number of individual doses that are administered during the course of that day. In the case of an immediate release parenteral formulation administration may be continuous (for example by way of infusion). A preferred daily oral dose is 20-500 mg and a preferred parenteral dose is in the range 0.1-50 mg.  
      General Procedures  
      TLC was performed on silica gel. Chiral HPLC analysis was performed using a 46 mm×250 mm Chiralcel OD column with a 5 cm guard column. The column temperature was maintained at 35° C. A flow rate of 1.0 mL/min was used. A Gilson 115 UV detector at 228 nm was used. The mobile phase consisted of hexanes, ethanol and trifluroacetic acid and the appropriate ratios are listed for each compound. Typically, the product was dissolved in a minimal amount of ethanol and this was diluted with the mobile phase.  
      In Preparations A to I below, LC-MS/MS was performed using a HP-1100 instrument equipped with a CTC-PAL injector and a 5 Tm, 4×100 mm ThermoQuest, Hypersil BDS-C18 column. An API-3000 (Sciex) MS detector was used. The flow rate was 1.2 mL/min and the mobile phase (gradient) consisted of 10-90% acetonitrile with 90-10% of 4 mM aq. ammonium acetate, both containing 0.2% formic acid. Otherwise, low resolution mass spectra (LRMS) were recorded using a Micromass ZQ spectrometer in ESI posneg switching ion mode (mass range m/z 100-800); and high resolution mass spectra (HRMS) were recorded using a Micromass LCT spectrometer in ES negative ionization mode (mass range m/z 100-1000) with Leucine Enkephalin (C 28 H 37 N 5 O 7 ) as internal mass standard.  
       1 H NMR spectra were recorded using tetramethylsilane as the internal standard.  
      Processes for the synthesis of compounds of formula (I) are contained in International Patent Application No. PCT/SE01/02657 (WO 02/44145, earliest priority date 1 Dec. 2000, filed 30 Nov. 2001, published 6 Jun. 2002)), relevant information from which is incorporated herein.  
      Preparation A: Preparation of Compound A  
     (i) 3-Chloro-5-methoxybenzaldehyde  
      3,5-Dichloroanisole (74.0 g, 419 mmol) in THF (200 mL) was added dropwise to magnesium metal (14.2 g, 585 mmol, pre-washed with 0.5 N HCl) in THF (100 mL) at 25° C. After the addition, 1,2-dibromoethane (3.9 g, 20.8 mmol) was added dropwise. The resultant dark brown mixture was heated at reflux for 3 h. The mixture was cooled to 0° C., and N,N-dimethylformamide (60 mL) was added in one portion. The mixture was partitioned with diethyl ether (3×400 mL) and 6N HCl (500 mL). The combined organic extracts were washed with brine (300 mL), dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give an oil. Flash chromatography (2×) on silica gel eluting with Hex:EtOAc (4:1) afforded the sub-title compound (38.9 g, 54%) as a yellow oil.  
       1 H NMR (300 MHz, CDCl 3 ) δ 9.90 (s, 1H), 7.53 (s, 1H), 7.38 (s, 1H), 7.15 (s, 1H), 3.87 (s, 3H).  
     (ii) 3-Chloro-5-hydroxybenzaldehyde  
      A solution of 3-chloro-5-methoxybenzaldehyde (22.8 g, 134 mmol; see step (i) above) in CH 2 Cl 2  (250 mL) was cooled to 0° C. Boron tribromide (15.8 mL, 167 mmol) was added dropwise over 15 min. After stirring, the reaction mixture for 2 h, H 2 O (50 mL) was added slowly. The solution was then extracted with Et 2 O (2×100 mL). The organic layers were combined, dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with Hex:EtOAc (4:1) afforded the sub-title compound (5.2 g, 25%).  
       1 H NMR (300 MHz, CDCl 3 ) δ 9.85 (s, 1H), 7.35 (s, 1H), 7.20 (s, 1H), 7.10 (s, 1H), 3.68 (s, 1H)  
     (iii) 3-Chloro-5-difluoromethoxybenzaldehyde  
      A solution of 3-chloro-5-hydroxybenzaldehyde (7.5 g, 48 mmol; see step (ii) above) in 2-propanol (250 mL) and 30% KOH (100 mL) was heated to reflux. While stirring, CHClF 2  was bubbled into the reaction mixture for 2 h. The reaction mixture was cooled, acidified with 1N HCl and extracted with EtOAc (2×100 mL). The organics were washed with brine (100 mL), dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with Hex:EtOAc (4:1) afforded the sub-title compound (4.6 g, 46%).  
       1 H NMR (300 MHz, CDCl 3 ) δ 9.95 (s, 1H), 7.72 (s, 1H), 7.52 (s, 1H), 7.40 (s, 1H), 6.60 (t, J H-F =71.1 Hz, 1H)  
     (iv) Ph(3-Cl)(5-OCHF 2 )—(R,S)CH(OTMS)CN  
      A solution of 3-chloro-5-difluoromethoxybenzaldehyde (4.6 g, 22.3 mmol; see step (iii) above) in CH 2 Cl 2  (200 mL) was cooled to 0° C. ZnI 2  (1.8 g, 5.6 mmol) and trimethylsilyl cyanide (2.8 g, 27.9 mmol) were added and the reaction mixture was allowed to warm to room temperature and stirred for 15 h. The mixture was partially concentrated in vacuo yielding the sub-title compound as a liquid, which was used directly in step (v) below without further purification or characterization.  
     (v) Ph(3-Cl)(5-OCHF 2 )—(R,S)CH(OH)C(NH)OEt  
      Ph(3-Cl)(5-OCHF 2 )—(R,S)CH(OTMS)CN (6.82 g, assume 22.3 mmol; see step (iv) above) was added dropwise to HCl/EtOH (500 mL). The reaction mixture was stirred 15 h, then partially concentrated in vacuo yielding the sub-title compound as a liquid, which was used in step (vi) without further purification or characterization.  
     (vi) Ph(3-Cl)(5-OCHF 2 )—(R,S)CH(OH)C(O)OEt  
      Ph(3-Cl)(5-OCHF 2 )—(R,S)CH(OH)C(NH)OEt (6.24 g, assume 22.3 mmol; see step (v) above) was dissolved in THF (250 mL), 0.5M H 2 SO 4  (400 mL) was added and the reaction was stirred at 40° C. for 65 h, cooled and then partially concentrated in vacuo to remove most of the THF. The reaction mixture was then extracted with Et 2 O (3×100 mL), dried (Na 2 SO 4 ), filtered and concentrated in vacuo to afford the sub-title compound as a solid, which was used in step (vii) without further purification or characterization.  
     (vii) Ph(3-Cl)(5-OCHF 2 )—(R,S)CH(OH)C(O)OH  
      A solution of Ph(3-Cl)(5-OCHF 2 )—(R,S)CH(OH)C(O)OEt (6.25 g, assume 22.3 mmol; see step (vi) above) in 2-propanol (175 mL) and 20% KOH (350 mL) was stirred at room temperature 15 h. The reaction was then partially concentrated in vacuo to remove most of the 2-propanol. The remaining mixture was acidified with 1M H 2 SO 4 , extracted with Et 2 O (3×100 mL), dried (Na 2 SO 4 ) and concentrated in vacuo to give a solid. Flash chromatography on silica gel eluting with CHCl 3 :MeOH:concentrated NH 4 OH (6:3:1) afforded the ammonium salt of the sub-title compound. The ammonium salt was then dissolved in a mixture of EtOAc (75 mL) and H 2 O (75 mL) and acidified with 2N HCl. The organic layer was separated and washed with brine (50 mL), dried (Na 2 SO 4 ) and concentrated in vacuo to afford the sub-title compound (3.2 g, 57% from steps (iv) to (vii)).  
       1 H NMR (300 MHz, CD 3 OD) δ 7.38 (s, 1H), 7.22 (s, 1H), 7.15 (s, 1H), 6.89 (t, J H-F =71.1 Hz, 1H), 5.16 (s, 1H)  
     (viii) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)OH (a) and Ph(3-Cl)(5-OCHF 2 )—(S)CH(OAc)C(O)OH (b)  
      A mixture of Ph(3-Cl)(5-OCHF 2 )—(R,S)CH(OH)C(O)OH (3.2 g, 12.7 mmol; see step (vii) above) and Lipase PS “Amano” (˜2.0 g) in vinyl acetate (125 mL) and MTBE (125 mL) was heated at reflux for 48 h. The reaction mixture was cooled, filtered through Celite® and the filter cake washed with EtOAc. The filtrate was concentrated in vacuo and subjected to flash chromatography on silica gel eluting with CHCl 3 :MeOH:concentrated NH 4 OH (6:3:1) yielding the ammonium salts of the sub-title compounds (a) and (b). Compound (a) as a salt was dissolved in H 2 O, acidified with 2N HCl and extracted with EtOAc. The organic layer was washed with brine, dried (Na 2 SO 4 ), filtered and concentrated in vacuo to afford the sub-title compound (a) (1.2 g, 37%).  
      For Sub-Title Compound (a)  
       1 H NMR (300 MHz, CD 3 OD) δ 7.38 (s, 1H), 7.22 (s, 1H), 7.15 (s, 1H), 6.89 (t, J H-F =71.1 Hz, 1H), 5.17 (s, 1H)  
     (ix) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(Teoc)  
      To a solution of Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)OH (1.1 g, 4.4 mmol; see step (viii) above) and H-Aze-Pab(Teoc) (see international patent application WO 00/42059, 2.6 g, 5.7 mmol) in DMF (50 mL) at 0° C. was added PyBOP (2.8 g, 5.3 mmol) and collidine (1.3 g, 10.6 mmol). The reaction was stirred at 0° C. for 2 h and then at room temperature for an additional 15 h. The reaction mixture was concentrated in vacuo and flash chromatographed on silica gel (3×), eluting first with CHCl 3 :EtOH (9:1), then with EtOAc:EtOH (20:1) and finally eluting with CH 2 Cl 2 :CH 3 OH (95:5) to afford the sub-title compound (1.0 g, 37%) as a white solid.  
       1 H NMR (300 MHz, CD 3 OD, mixture of rotamers) δ 7.79-7.85 (d, J=8.7 Hz, 2H), 7.15-7.48 (m, 5H), 6.89 and 6.91 (t, J H-F =71.1 Hz, 1H), 5.12 and 5.20 (s, 1H), 4.75-4.85 (m, 1H), 3.97-4.55 (m, 6H), 2.10-2.75 (m, 2H), 1.05-1.15 (m, 2H), 0.09 (s, 9H)  
      MS (m/z) 611 (M+1) +   
     (x) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(OMe, Teoc)  
      Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(Teoc) (0.40 g, 0.65 mmol; see step (ix) above), was dissolved in 20 mL of acetonitrile and 0.50 g (6.0 mmol) of O-methyl hydroxylamine hydrochloride was added. The mixture was heated at 70° C. for 2 h. The solvent was evaporated and the residue was partitioned between water and ethyl acetate. The aqueous phase was extracted twice more with ethyl acetate and the combined organic phase was washed with water, brine, dried (Na 2 SO 4 ), filtered and evaporated. Yield: 0.41 g (91%).  
       1 H-NMR (400 MHz; CDCl 3 ): δ 7.83 (bt, 1H), 7.57 (bs, 1H), 7.47 (d, 2H), 7.30 (d, 2H), 7.20 (m, 1H), 7.14 (m, 1H), 7.01 (m, 1H), 6.53 (t, 1H), 4.89 (s, 1H), 4.87 (m, 1H), 4.47 (m, 2H), 4.4-4.2 (b, 1H), 4.17-4.1 (m, 3H), 3.95 (s, 3H), 3.67 (m, 1H), 2.68 (m, 1H), 2.42 (m, 1H) 0.97 (m, 2H), 0.01 (s, 9H).  
     (xi) Compound A  
      Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(OMe, Teoc) (0.40 g, 0.62 mmol; see step (x) above), was dissolved in 5 mL of TFA and allowed to react for 30 min. TFA was evaporated and the residue was partitioned between ethyl acetate and NaHCO 3  (aq.). The aqueous phase was extracted twice more with ethyl acetate and the combined organic phase was washed with water, brine, dried (Na 2 SO 4 ), filtered and evaporated. The product was freeze dried from water/acetonitrile. No purification was necessary. Yield: 0.28 g (85%).  
       1 H-NMR (600 MHz; CDCl 3 ): δ 7.89 (bt, 1H), 7.57 (d, 2H), 7.28 (d, 2H), 7.18 (m, 1H), 7.13 (m, 1H), 6.99 (m, 1H), 6.51 (t, 1H), 4.88 (s, 1H), 4.87 (m, 1H), 4.80 (bs, 2H), 4.48 (dd, 1H), 4.43 (dd, 1H), 4.10 (m, 1H), 3.89 (s, 3H), 3.68 (m, 1H), 2.68 (m, 1H), 2.40 (m, 1H).  
       13 C-NMR (125 MHz; CDCl 3 ): (carbonyl and/or amidine carbons, rotamers) δ 172.9, 170.8, 152.7, 152.6  
      HRMS calculated for C 22 H 23 ClF 2 N 4 O 5  (M−H) −  495.1242, found 495.1247  
      Preparation B: Preparation of Compound B  
     (i) 2,6-Difluoro-4[(methylsulfinyl)(methylthio)methyl]benzonitrile  
      (Methylsulfinyl)(methylthio)methane (7.26 g, 0.0584 mol) was dissolved in 100 mL of dry THF under argon and was cooled to −78° C. Butyllithium in hexane (16 mL 1.6M, 0.0256 mol) was added dropwise with stirring. The mixture was stirred for 15 min. Meanwhile, a solution of 3,4,5-trifluorobenzonitrile (4.0 g, 0.025 mmol) in 100 mL of dry THF was cooled to −78° C. under argon and the former solution was added through a cannula to the latter solution over a period of 35 min. After 30 min, the cooling bath was removed and when the reaction had reached room temperature it was poured into 400 mL of water. The THF was evaporated and the remaining aqueous layer was extracted three times with diethyl ether. The combined ether phase was washed with water, dried (Na 2 SO 4 ) and evaporated. Yield: 2.0 g(30%).  
       1 H NMR (500 MHz, CDCl 3 ) δ 7.4-7.25 (m, 2H), 5.01 (s, 1H, diasteromer), 4.91 (s, 1H, diasteromer), 2.88 (s, 3H, diasteromer), 2.52 (s, 3H, diasteromer), 2.49 (s, 3H, diasteromer), 2.34 (s, 3H, diasteromer), 1.72 (broad, 1H)  
     (ii) 2,6-Difluoro-4-formylbenzonitrile  
      2,6-Difluoro-4[(methylsulfinyl)(methylthio)methyl]benzonitrile (2.17 g, 8.32 mmol; see step (i) above) was dissolved in 90 mL of THF and 3.5 mL of concentrated sulfuric acid was added. The mixture was left at room temperature for 3 days and subsequently poured into 450 mL of water. Extraction three times with EtOAc followed and the combined ethereal phase was washed twice with aqueous sodium bicarbonate and with brine, dried (Na 2 SO 4 ) and evaporated. Yield: 1.36 g (98%). The position of the formyl group was established by  13 C NMR. The signal from the fluorinated carbons at 162.7 ppm exhibited the expected coupling pattern with two coupling constants in the order of 260 Hz and 6.3 Hz respectively corresponding to an ipso and a meta coupling from the fluorine atoms.  
       1 H NMR (400 MHz, CDCl 3 ) δ 10.35 (s, 1H), 7.33 (m, 2H)  
     (iii) 2,6-Difluoro-4-hydroxymethylbenzonitrile  
      2,6-Difluoro-4-formylbenzonitrile (1.36 g, 8.13 mmol; see step (ii) above) was dissolved in 25 mL of methanol and cooled on an ice bath. Sodium borohydride (0.307 g, 8.12 mmol) was added in portions with stirring and the reaction was left for 65 min. The solvent was evaporated and the residue was partitioned between diethyl ether and aqueous sodium bicarbonate. The ethereal layer was washed with more aqueous sodium bicarbonate and brine, dried (Na 2 SO 4 ) and evaporated. The crude product crystallised soon and could be used without further purification. Yield: 1.24 g (90%).  
       1 H NMR (400 MHz, CDCl 3 ) δ 7.24 (m, 2H), 4.81 (s, 2H), 2.10 (broad, 1H)  
     (iv) 4-Cyano-2,6-difluorobenzyl methanesulfonate  
      To an ice cooled solution of 2,6-difluoro-4-hydroxymethylbenzonitrile (1.24 g, 7.32 mmol; see step (iii) above) and methanesulfonyl chloride (0.93 g, 8.1 mmol) in 60 mL of methylene chloride was added triethylamine (0.81 g, 8.1 mmol) with stirring. After 3 h at 0° C., the mixture was washed twice with 1M HCl and once with water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification. Yield: 1.61 g (89%).  
       1 H NMR (300 MHz, CDCl 3 ) δ 7.29 (m, 2H), 5.33 (s, 2H), 3.07 (s, 3H)  
     (v) 4-Azidomethyl-2,6-difluorobenzonitrile  
      A mixture of 4-cyano-2,6-difluorobenzyl methanesulfonate (1.61 g, 6.51 mmol; see step (iv) above) and sodium azide (0.72 g, 0.0111 mol) in 10 mL of water and 20 mL of DMF was stirred at room temperature overnight. The resultant was subsequently poured into 200 mL of water and extracted three times with diethyl ether. The combined ethereal phase was washed five times with water, dried (Na 2 SO 4 ) and evaporated. A small sample was evaporated for NMR purposes and the product crystallised. The rest was evaporated cautiously but not until complete dryness. Yield (theoretically 1.26 g) was assumed to be almost quantitative based on NMR and analytical HPLC.  
       1 H NMR (400 MHz, CDCl 3 ) δ 7.29 (m, 2H), 4.46 (s, 2H)  
     (vi) 4-Aminomethyl-2,6-difluorobenzonitrile  
      This reaction was carried out according to the procedure described in  J. Chem. Res . ( M ) (1992) 3128. To a suspension of 520 mg of 10% Pd/C (50% moisture) in 20 mL of water was added a solution of sodium borohydride (0.834 g, 0.0221 mol) in 20 mL of water. Some gas evolution resulted. 4-Azidomethyl-2,6-difluorobenzonitrile (1.26 g, 6.49 mmol; see step (v) above) was dissolved in 50 mL of THF and added to the aqueous mixture on an ice bath over 15 min. The mixture was stirred for 4 h, whereafter 20 mL of 2M HCl was added and the mixture was filtered through Celite. The Celite was rinsed with more water and the combined aqueous phase was washed with EtOAc and subsequently made alkaline with 2M NaOH. Extraction three times with methylene chloride followed and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. Yield: 0.87 g (80%).  
       1 H NMR (400 MHz, CDCl 3 ) δ 7.20 (m, 2H), 3.96 (s, 2H), 1.51 (broad, 2H)  
     (vii) 2,6-Difluoro-4-tert-butoxycarbonylaminomethylbenzonitrile  
      A solution of 4-aminomethyl-2,6-difluorobenzonitrile (0.876 g, 5.21 mmol; see step (vi) above) was dissolved in 50 mL of THF and di-tert-butyl dicarbonate (1.14 g, 5.22 mmol) in 10 mL of THF was added. The mixture was stirred for 3.5 h. The THF was evaporated and the residue was partitioned between water and EtOAc. The organic layer was washed three times with 0.5 M HCl and water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification. Yield: 1.38 g (99%).  
       1 H NMR (300 MHz, CDCl 3 ) δ 7.21 (m, 2H), 4.95 (broad, 1H), 4.43 (broad, 2H), 1.52 (s, 9H)  
     (viii) Boc-Pab(2,6-diF)(OH)  
      A mixture of 2,6-difluoro-4-tert-butoxycarbonylaminomethylbenzonitrile (1.38 g, 5.16 mmol; see step (vii) above), hydroxylamine hydrochloride (1.08 g, 0.0155 mol) and triethylamine (1.57 g, 0.0155 mol) in 20 mL of ethanol was stirred at room temperature for 36 h. The solvent was evaporated and the residue was partitioned between water and methylene chloride. The organic layer was washed with water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification. Yield: 1.43 g (92%).  
       1 H NMR (500 MHz, CD 3 OD) δ 7.14 (m, 2H), 4.97 (broad, 1H), 4.84 (broad, 2H), 4.40 (broad, 2H), 1.43 (s, 9H)  
     (ix) Boc-Pab(2,6-diF)xHOAc  
      This reaction was carried out according to the procedure described by Judkins et al,  Synth. Comm.  (1998) 4351. Boc-Pab(2,6-diF)(OH) (1.32 g, 4.37 mmol; see step (viii) above), acetic anhydride (0.477 g, 4.68 mmol) and 442 mg of 10% Pd/C (50% moisture) in 100 mL of acetic acid was hydrogenated at 5 atm pressure for 3.5 h. The mixture was filtered through Celite, rinsed with ethanol and evaporated. The residue was freeze-dried from acetonitrile and water and a few drops of ethanol. The sub-title product could be used without further purification. Yield: 1.49 g (99%).  
       1 H NMR (400 MHz, CD 3 OD) δ 7.45 (m, 2H), 4.34 (s, 2H), 1.90 (s, 3H), 1.40 (s, 9H)  
     (x) Boc-Pab(2,6-diF)(Teoc)  
      To a solution of Boc-Pab(2,6-diF)xHOAc (1.56 g, 5.49 mmol; see step (ix) above) in 100 mL of THF and 1 mL of water was added 2-(trimethylsilyl)ethyl p-nitrophenyl carbonate (1.67 g, 5.89 mmol). A solution of potassium carbonate (1.57 g, 0.0114 mol) in 20 mL of water was added dropwise over 5 min. The mixture was stirred overnight. The THF was evaporated and the residue was partitioned between water and methylene chloride. The aqueous layer was extracted with methylene chloride and the combined organic phase was washed twice with aqueous sodium bicarbonate, dried (Na 2 SO 4 ) and evaporated. Flash chromatography on silica gel with heptane/EtOAc=2/1 gave 1.71 g (73%) of pure compound.  
       1 H NMR (400 MHz, CDCl 3 ) δ 7.43 (m, 2H), 4.97 (broad, 1H), 4.41 (broad, 2H), 4.24 (m, 2H), 1.41 (s, 9H), 1.11 (m, 2H), 0.06 (s, 9H)  
     (xi) Boc-Aze-Pab(2,6-diF)(Teoc)  
      Boc-Pab(2,6-diF)(Teoc) (1.009 g, 2.35 mmol; see step (x) above) was dissolved in 50 mL of EtOAc saturated with HCl(g). The mixture was left for 10 min., evaporated and dissolved in 18 mL of DMF, and then cooled on an ice bath. Boc-Aze-OH (0.450 g, 2.24 mmol), PyBOP (1.24 g, 2.35 mmol) and lastly diisopropylethyl amine (1.158 g, 8.96 mmol) were added. The reaction mixture was stirred for 2 h and then poured into 350 mL of water and extracted three times with EtOAc. The combined organic phase was washed with brine, dried (Na 2 SO 4 ) and evaporated. Flash chromatography on silica gel with heptane:EtOAc (1:3) gave 1.097 g (96%) of the desired compound.  
       1 H NMR (500 MHz, CDCl 3 ) δ 7.46 (m, 2H), 4.65-4.5 (m, 3H), 4.23 (m, 2H), 3.87 (m, 1H), 3.74 (m, 1H), 2.45-2.3 (m, 2H), 1.40 (s, 9H), 1.10 (m, 2H), 0.05 (s, 9H)  
     (xii) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(2,6-diF)(Teoc)  
      Boc-Aze-Pab(2,6-diF)(Teoc) (0.256 g, 0.500 mmol; see step (xi) above) was dissolved in 20 mL of EtOAc saturated with HCl(g). The mixture was left for 10 min. and evaporated and dissolved in 5 mL of DMF. Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)OH (0.120 g, 0.475 mmol; see Preparation A(viii) above), PyBOP (0.263 g, 0.498 mmol) and lastly diisopropylethyl amine (0.245 g, 1.89 mmol) were added. The reaction mixture was stirred for 2 h and then poured into 350 mL of water and extracted three times with EtOAc. The combined organic phase was washed with brine, dried (Na 2 SO 4 ) and evaporated. Flash chromatography on silica gel with EtOAc gave 0.184 g (60%) of the desired sub-title compound.  
       1 H NMR (400 MHz, CD 3 OD, mixture of rotamers) δ 7.55-7.45 (m, 2H), 7.32 (m, 1H, major rotamer), 7.27 (m, 1H, minor rotamer), 7.2-7.1 (m, 2H), 6.90 (t, 1H, major rotamer), 6.86 (t, 1H, minor rotamer), 5.15 (s, 1H, major rotamer), 5.12 (m, 1H, minor rotamer), 5.06 (s, 1H, minor rotamer), 4.72 (m, 1H, major rotamer), 4.6-4.45 (m, 2H), 4.30 (m, 1H, major rotamer), 4.24 (m, 2H), 4.13 (m, 1H, major rotamer), 4.04 (m, 1H, minor rotamer), 3.95 (m, 1H, minor rotamer), 2.62 (m, 1H, minor rotamer), 2.48 (m, 1H, major rotamer), 2.22 (m, 1H, major rotamer), 2.10 (m, 1H, minor rotamer), 1.07 (m, 2H), 0.07 (m, 9H)  
     (xiii) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(2,6-diF)(OMe,Teoc)  
      A mixture of Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(2,6-diF)(Teoc) (64 mg, 0.099 mmol; see step (xii) above) and O-methyl hydroxylamine hydrochloride (50 mg, 0.60 mmol) in 4 mL of acetonitrile was heated at 70° C. for 3 h. The solvent was evaporated and the residue was partitioned between water and EtOAc. The aqueous layer was extracted twice with EtOAc and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification. Yield: 58 mg (87%).  
       1 H NMR (400 MHz, CDCl 3 ) δ 7.90 (bt, 1H), 7.46 (m, 1H), 7.25-6.95 (m, 5H), 6.51, t, 1H), 4.88 (s, 1H), 4.83 (m, 1H), 4.6-4.5 (m, 2H), 4.4-3.9 (m, 4H), 3.95 (s, 3H), 3.63 (m, 1H), 2.67 (m, 1H), 2.38 (m, 1H), 1.87 (broad, 1H), 0.98 (m, 2H), 0.01, s, 9H)  
     (xiv) Compound B  
      Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(2,6-diF)(OMe,Teoc) (58 mg, 0.086 mmol; see step (xiii) above) was dissolved in 3 mL of TFA, cooled on an ice bath and allowed to react for 2 h. The TFA was evaporated and the residue dissolved in EtOAc. The organic layer was washed twice with aqueous sodium carbonate and water, dried (Na 2 SO 4 ) and evaporated. The residue was freeze-dried from water and acetonitrile to give 42 mg (92%) of the title compound.  
       1 H NMR (300 MHz, CDCl 3 ) δ 7.95 (bt, 1H), 7.2-7.1 (m, 4H), 6.99 (m, 1H), 6.52 (t, 1H), 4.88 (s, 1H), 4.85-4.75 (m, 3H), 4.6-4.45 (m, 2H), 4.29 (broad, 1H), 4.09 (m, 1H), 3.89 (s, 3H), 3.69 (m, 1H), 2.64 (m, 1H), 2.38 (m, 1H), 1.85 (broad, 1H)  
       13 C-NMR (100 MHz; CDCl 3 ): (carbonyl and/or amidine carbons) δ 172.1, 169.8, 151.9  
      APCI-MS: (M+1)=533/535 m/z  
      Preparation C: Preparation of Compound C  
     (i) (2-Monofluoroethyl) methanesulfonate  
      To a magnetically stirred solution of 2-fluoroethanol (5.0 g, 78.0 mmol) in CH 2 Cl 2  (90 mL) under nitrogen at 0° C. was added triethylamine (23.7 g, 234 mmol) and methanesulfonyl chloride (10.7 g, 93.7 mmol). The mixture was stirred at 0° C. for 1.5 h, diluted with CH 2 Cl 2  (100 mL) and washed with 2N HCl (100 mL). The aqueous layer was extracted with CH 2 Cl 2  (50 mL) and the combined organic extracts washed with brine (75 mL), dried (Na 2 SO 4 ), filtered and concentrated in vacuo to afford the sub-title compound (9.7 g, 88%) as a yellow oil which was used without further purification.  
       1 H NMR (300 MHz, CDCl 3 ) δ 4.76 (t, J=4 Hz, 1H), 4.64 (t, J=4 Hz, 1H), 4.52 (t, J=4 Hz, 1H), 4.43 (t, J=4 Hz, 1H), 3.09 (s, 3H).  
     (ii) 3-Chloro-5-monofluoroethoxybenzaldehyde  
      To a solution of 3-chloro-5-hydroxybenzaldehyde (8.2 g, 52.5 mmol; see Preparation A(ii) above) and potassium carbonate (9.4 g, 68.2 mmol) in DMF (10 mL) under nitrogen was added a solution of (2-monofluoroethyl) methanesulfonate (9.7 g, 68.2 mmol; see step (i) above) in DMF (120 mL) dropwise at room temperature. The mixture was heated to 1001° C. for 5 h and then stirred overnight at room temperature. The reaction was cooled to 0° C., poured into ice-cold 2N HCl and extracted with EtOAc. The combined organic extracts were washed with brine, dried (Na 2 SO 4 ), filtered and concentrated in vacuo. The brown oil was chromatographed on silica gel eluting with Hex:EtOAc (4:1) to afford the sub-title compound (7.6 g, 71%) as a yellow oil.  
       1 H NMR (300 MHz, CDCl 3 ) δ 9.92 (s, 1H), 7.48 (s, 1H), 7.32 (s, 1H), 7.21 (s, 1H), 4.87 (t, J=4 Hz, 1H), 4.71 (t, J=3 Hz, 1H), 4.33 (t, J=3 Hz, 1H), 4.24 (t, J=3 Hz, 1H).  
     (iii) Ph(3-Cl)(5-OCH 2 CH 2 F)—(R,S)CH(OTMS)CN  
      To a solution of 3-chloro-5-monofluoroethoxybenzaldehyde (7.6 g, 37.5 mmol; see step (ii) above) and zinc iodide (3.0 g, 9.38 mmol) in CH 2 Cl 2  (310 mL) was added trimethylsilyl cyanide (7.4 g, 75.0 mmol) dropwise at 0° C. under nitrogen. The mixture was stirred at 0° C. for 3 h and at room temperature overnight. The reaction was diluted with H 2 O (300 mL), the organic layer was separated, dried (Na 2 SO 4 ), filtered and concentrated in vacuo to afford the sub-title compound (10.6 g, 94%) as a brown oil that was used without further purification or characterisation.  
     (iv) Ph(3-Cl)(5-OCH 2 CH 2 F)—(R,S)CH(OH)C(O)OH  
      Concentrated hydrochloric acid (100 mL) was added to Ph(3-Cl)(5-OCH 2 CH 2 F)—(R,S)CH(OTMS)CN (10.6 g, 5.8 mmol; see step (iii) above) and the solution stirred at 100° C. for 3 h. After cooling to room temperature, the reaction was further cooled to 0° C., basified slowly with 3N NaOH (300 mL) and washed with Et 2 O (3×200 mL). The aqueous layer was acidified with 2N HCl (80 mL) and extracted with EtOAc (3×300 mL). The combined EtOAc extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to afford the sub-title compound (8.6 g, 98%) as a pale yellow solid that was used without further purification.  
      R f =0.28 (90:8:2 CHCl 3 :MeOH:concentrated NH 4 OH)  
       1 H NMR (300 MHz, CD 3 OD) δ 7.09 (s, 1H), 7.02 (s, 1H), 6.93 (s, 1H), 5.11 (s, 1H), 4.77-4.81 (m, 1H), 4.62-4.65 (m, 1H), 4.25-4.28 (m, 1H), 4.15-4.18 (m, 1H).  
     (v) Ph(3-Cl)(5-OCH 2 CH 2 F)—(S)CH(OAc)C(O)OH (a) and Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)OH (b)  
      A solution of Ph(3-Cl)(5-OCH 2 CH 2 F)—(R,S)CH(OH)C(O)OH (8.6 g, 34.5 mmol; see step (iv) above) and Lipase PS “Amano” (4.0 g) in vinyl acetate (250 mL) and MTBE (250 mL) was heated at 70° C. under nitrogen for 3 d. The reaction was cooled to room temperature and the enzyme removed by filtration through Celite®. The filter cake was washed with EtOAc and the filtrate concentrated in vacuo. Chromatography on silica gel eluting with CHCl 3 :MeOH:Et 3 N (90:8:2) afforded the triethylamine salt of sub-title compound (a) as a yellow oil. In addition, the triethylamine salt of sub-title compound (b) (4.0 g) was obtained. The salt of sub-title compound (b) was dissolved in H 2 O (250 mL), acidified with 2N HCl and extracted with EtOAc (3×200 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to yield the sub-title compound (b) (2.8 g, 32%) as a yellow oil.  
      Data for Sub-Title Compound (b):  
     
         
         
           
              R f =0.28 (90:8:2 CHCl 3 :MeOH:concentrated NH 4 OH)  
           
         
       
    
       1 H NMR (300 MHz, CD 3 OD) δ 7.09 (s, 1H), 7.02 (s, 1H), 6.93 (s, 1H), 5.11 (s, 1H), 4.77-4.81 (m, 1H), 4.62-4.65 (m, 1H), 4.25-4.28 (m, 1H), 4.15-4.18 (m, 1H).  
     (vi) Compound C  
      To a solution of Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)OH (818 mg, 3.29 mmol; see step (v) above) in DMF (30 mL) under nitrogen at 0° C. was added HAze-Pab(OMe).2HCl (1.43 g, 4.27 mmol, see international patent application WO 00/42059), PyBOP (1.89 g, 3.68 mmol), and DIPEA (1.06 g, 8.23 mmol). The reaction was stirred at 0° C. for 2 h and then at room temperature overnight. The mixture was concentrated in vacuo and the residue chromatographed two times on silica gel, eluting first with CHCl 3 :EtOH (15:1) and second with EtOAc:EtOH (20:1) to afford the title compound (880 mg, 54%).  
      R f =0.60 (10:1 CHCl 3 :EtOH)  
       1 H NMR (300 MHz, CD 3 OD, complex mixture of rotamers) δ 7.58-7.60 (d, J=8 Hz, 2H), 7.34 (d, J=7 Hz, 2H), 7.05-7.08 (m, 2H), 6.95-6.99 (m, 1H), 5.08-5.13 (m, 1H), 4.77-4.82 (m, 1H), 4.60-4.68 (m, 1H), 3.99-4.51 (m, 7H), 3.82 (s, 3H), 2.10-2.75 (m, 2H).  
       13 C-NMR (150 MHz; CD 3 OD): (carbonyl and/or amidine carbons) δ 173.3, 170.8, 152.5.  
      APCI-MS: (M+1)=493 m/z.  
     Preparation of Compound D (Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab) Compound D  
      Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(Teoc) (0.045 g, 0.074 mmol; see Preparation A (ix) above), was dissolved in 3 mL of TFA and allowed to react for 1 h. TFA was evaporated and the residue was freeze dried from water/acetonitrile to yield 0.043 g (100%) of the sub-title compound as its TFA salt.  
       1 H-NMR (400 MHz; CD 3 OD) rotamers: δ 7.8-7.75 (m, 2H), 7.55-7.5 (m, 2H), 7.35 (m, 1H, major rotamer), 7.31 (m, 1H, minor rotamer), 7.19 (m, 1H, major rotamer), 7.15 (m, 1H), 7.12 (m, 1H, minor rotamer), 6.89 (t, 1H, major rotamer), 6.87 (t, 1H, minor rotamer), 5.22 (m, 1H, minor rotamer), 5.20 (s, 1H, major rotamer), 5.13 (s, 1H, minor rotamer), 4.80 (m, 1H, major rotamer), 4.6-4.4 (m, 2H), 4.37 (m, 1H, major rotamer), 4.19 (m, 1H, major rotamer), 4.07 (m, 1H, minor rotamer), 3.98 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.55 (m, 1H, major rotamer), 2.29 (m, 1H, major rotamer), 2.15 (m, 1H, minor rotamer)  
       13 C-NMR (100 MHz; CD 3 OD): (carbonyl and/or amidine carbons, rotamers) δ 172.6, 172.5, 172.0, 171.7, 167.0  
      MS (m/z) 465 (M−1) − , 467 (M+1) +   
     Preparation of Compound E (Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(2,6-diF)) Compound E  
      Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(2,6-diF)(Teoc) (81 mg, 0.127 mmol; see Preparation B (xii) above) was dissolved in 0.5 mL of methylene chloride and cooled on an ice bath. TFA (3 mL) was added and the reaction was left for 75 min. The TFA was evaporated and the residue was freeze dried from water and acetonitrile. The crude product was purified by preparative RPLC with CH 3 CN:0.1M NH 4 OAc (35:65) to produce 39 mg (55%) of the title compound as its HOAc salt, purity: 99%.  
       1 H NMR (400 MHz, CD 3 OD mixture of rotamers) δ 7.5-7.4 (m, 2H), 7.32 (m, 1H, major rotamer), 7.28 (m, 1H, minor rotamer), 7.2-7.1 (m, 3H) 6.90 (t, 1H, major rotamer), 6.86 (t, minor rotamer), 5.15 (s, 1H, major rotamer), 5.14 (m, 1H, minor rotamer), 5.07 (s, 1H, minor rotamer), 4.72 (m, 1H, major rotamer), 4.65-4.45 (m, 2H), 4.30 (m, 1H, major rotamer), 4.16 (m, 1H, major rotamer), 4.03 (m, 1H, minor rotamer), 3.95 (m, 1H, minor rotamer), 2.63 (m, 1H, minor rotamer), 2.48 (m, 1H, major rotamer), 2.21 (m, 1H, major rotamer), 2.07 (m, 1H, minor rotamer), 1.89 (s, 3H)  
       13 C-NMR (75 MHz; CD 3 OD): (carbonyl and/or amidine carbons, mixture of rotamers) δ 171.9, 171.2, 165.0, 162.8, 160.4  
      APCI-MS: (M+1)=503/505 m/z.  
     Preparation of Compound F (Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)-Aze-PabxTFA)  
     (i) Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)-Aze-Pab(Teoc)  
      To a solution of Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)OH (940 mg, 3.78 mmol; see Preparation C (v) above) in DMF (30 mL) under nitrogen at 0° C. was added HAze-Pab(Teoc)-HCl (2.21 g, 4.91 mmol), PyBOP (2.16 g, 4.15 mmol), and DIPEA (1.22 g, 9.45 mmol). The reaction was stirred at 0° C. for 2 h and then at room temperature for 4 h. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl 3 :EtOH (15:1) and second with EtOAc:EtOH (20:1) to afford the sub-title compound (450 mg, 20%) as a crushable white foam.  
      Mp: 80-88° C.  
      R f =0.60 (10:1 CHCl 3 :EtOH)  
       1 H NMR (300 MHz, CD 3 OD, complex mixture of rotamers) δ 7.79 (d, J=8 Hz, 2H), 7.42 (d, J=8 Hz, 2H), 7.05-7.08 (m, 1H), 6.93-6.99 (m, 2H), 5.08-5.13 (m, 1H), 4.75-4.80 (m, 2H), 4.60-4.68 (m, 1H), 3.95-4.55 (m, 8H), 2.10-2.75 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s, 9H).  
      APCI-MS: (M+1)=607 m/z.  
     (ii) Compound F  
      Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)-Aze-Pab(Teoc) (0.357 g, 0.589 mmol; see step (i) above), was dissolved in 10 mL of TFA and allowed to react for 40 min. TFA was evaporated and the residue was freeze dried from water/acetonitrile to yield 0.33 g (93%) of the title compound as its TFA salt.  
       1 H-NMR (600 MHz; CD 3 OD) rotamers: δ 7.8-7.7 (m, 2H), 7.54 (d, 2H), 7.08 (s, 1H, major rotamer), 7.04 (s, 1H, minor rotamer), 6.99 (s, 1H, major rotamer), 6.95 (s, 1H), 6.92 (s, 1H, minor rotamer), 5.18 (m, 1H, minor rotamer), 5.14 (s, 1H, major rotamer), 5.08 (s, 1H, minor rotamer), 4.80 (m, 1H, major rotamer), 4.73 (m, 1H), 4.65 (m, 1H), 4.6-4.4 (m, 2H), 4.35 (m, 1H, major rotamer), 4.21 (doublet of multiplets, 2H), 4.12 (m, 1H, major rotamer), 4.06 (m, 1H, minor rotamer), 3.99 (m, 1H, minor rotamer), 2.69 (m, 1H, minor rotamer), 2.53 (m, 1H, major rotamer), 2.29 (m, 1H, major rotamer), 2.14 (m, 1H, minor rotamer).  
       13 C-NMR (150 MHz; CD 3 OD): (carbonyl and/or amidine carbons) δ 172.8, 172.1, 167.4.  
      ESI-MS+: (M+1)=463 (m/z)  
     Preparation of Compound G (Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(OH))  
     (i) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(OH, Teoc)  
      Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(Teoc) (0.148 g, 0.24 mmol; see Preparation A step (ix) above), was dissolved in 9 mL of acetonitrile and 0.101 g (1.45 mmol) of hydroxylamine hydrochloride was added. The mixture was heated at 70° C. for 2.5 h, filtered through Celite® and evaporated. The crude product (0.145 g; 75% pure) was used directly in the next step without further purification.  
     (ii) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(OH)  
      Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)-Aze-Pab(OH, Teoc) (0.145 g, 0.23 mmol; see step (i) above), was dissolved in 0.5 mL of CH 2 Cl 2  and 9 mL of TFA. The reaction was allowed to proceed for 60 minutes. TFA was evaporated and the residue was purified using preparative HPLC. The fractions of interest were pooled and freeze-dried (2×), yielding 72 mg (yield over two steps 62%) of the title compound.  
      MS (m/z) 482 (M−1) − ; 484 (M+1) +   
       1 H-NMR (400 MHz; CD 3 OD): δ 7.58 (d, 2H), 7.33 (m, 3H), 7.15 (m, 2H), 6.89 (t, 1H major rotamer), 6.86 (t, 1H minor rotamer), 5.18 (s, 1H major rotamer; and m, 1H minor rotamer), 5.12 (s, 1H minor rotamer), 4.77 (m, 1H major rotamer), 4.42 (m, 2H), 4.34 (m, 1H major rotamer), 4.14 (m, 1H major rotamer), 4.06 (m, 1H minor rotamer), 3.95 (m, 1H minor rotamer), 2.66 (m, 1H minor rotamer), 2.50 (m, 1H major rotamer), 2.27 (m, 1H major rotamer), 2.14 (m, 1H minor rotamer)  
       13 C-NMR (100 MHz; CD 3 OD): (carbonyl and/or amidine carbons, rotamers) δ 172.4, 172.3, 172.0, 171.4 152.3, 152.1  
     Preparation of Compound H: Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-duF)(OH)  
     
       
         
         
             
             
         
       
     
     (i) Boc-(S)Aze-NHCH 2 -Ph(2,6-diF, 4-CN)  
      Boc-(S)Aze-OH (1.14 g, 5.6 mmol) was dissolved in 45 mL of DMF. 4-Aminomethyl-2,6-difluorobenzonitrile (1.00 g, 5.95 mol, see Example 1(xiv) above), PyBOP (3.10 g, 5.95 mmol) and DIPEA (3.95 mL, 22.7 mmol) were added and the solution was stirred at room temperature for 2 h. The solvent was evaporated and the residue was partitioned between H 2 O and EtOAc (75 mL each). The aqueous phase was extracted with 2×50 mL EtOAc and the combined organic phase was washed with brine and dried over Na 2 SO 4 . Flash chromatography (SiO 2 , EtOAc/heptane (3/1)) yielded the sub-title compound (1.52 g, 77%) as an oil which crystallized in the refrigerator.  
       1 H-NMR (400 MHz; CD 3 OD): δ 7.19 (m, 2H), 4.65-4.5 (m, 3H), 3.86 (m, 1H), 3.73 (m, 1H), 2.45-2.3 (m, 2H), 1.39 (s, 9H)  
     (ii) H—(S)Aze-NHCH 2 -Ph(2,6-diF, 4-CN)xHCl  
      Boc-(S)Aze-NHCH 2 -Ph(2,6-diF, 4-CN) (0.707 g, 2.01 mmol, see step (i) above) was dissolved in 60 mL of EtOAc saturated with HCl(g). After stirring at room temperature for 15 minutes, the solvent was evaporated. The residue was dissolved in CH 3 CN/H 2 O (1/1) and was freeze-dried to give the sub-title compound (0.567 g, 98%) as an off-white amorphous powder.  
       1 H-NMR (400 MHz; CD 3 OD): δ 7.49 (m, 2H), 4.99 (m, 1H), 4.58 (m, 2H), 4.12 (m, 1H), 3.94 (m, 1H), 2.80 (m, 1H), 2.47 (m, 1H)  
      MS (m/z) 252.0 (M+1) +   
     (iii) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-NHCH 2 -Ph(2,6-diF, 4-CN)  
      Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)OH (0.40 g, 1.42 mmol, see Example l(viii) above) was dissolved in 10 mL of DMF and H—(S)Aze-NHCH 2 -Ph(2,6-diF, 4-CN)xHCl (0.43 g, 1.50 mmol, see step (ii) above) and PyBOP (0.779 g, 1.50 mmol) were added, followed by DIPEA (1.0 mL, 5.7 mmol). After stirring at room temperature for 2 h, the solvent was evaporated. The residue was partitioned between H 2 O (200 mL) and EtOAc (75 mL). The aqueous phase was extracted with 2×75 mL EtOAc and the combined organic phase was washed with brine and dried over Na 2 SO 4 . Flash chromatography (SiO 2 , EtOAc/heptane (4/1)) yielded the sub-title compound (0.56 g, 81%) as an oil.  
       1 H-NMR (400 MHz; CD 3 OD) rotamers: δ 7.43 (m, 2H), 7.31 (m, 1H, major rotamer), 7.26 (m, 1H, minor rotamer), 7.2-7.1 (m, 2H), 6.90 (t, 1H, major rotamer), 6.86 (t, 1H, minor rotamer), 5.14 (s, 1H, major rotamer), 5.11 (m, 1H, minor rotamer), 5.04 (s, 1H, minor rotamer), 4.71 (m, 1H, major rotamer), 4.6-4.45 (m, 2H), 4.30 (m, 1H, major rotamer), 4.2-3.9 (m, 1H; and 1H, minor rotamer), 2.62 (m, 1H, minor rotamer), 2.48 (m, 1H, major rotamer), 2.21 (m, 1H, major rotamer), 2.09 (m, 1H, minor rotamer)  
       13 C-NMR (100 MHz; CD 3 OD): (carbonyl carbons) δ 171.9, 171.8  
      MS (m/z) 484.0, 485.9 (M−1) − , 486.0, 487.9 (M+1) +   
     (iv) Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF)(OH)  
      Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-NHCH 2 -Ph(2,6-diF, 4-CN) (0.555 g, 1.14 mmol, from step (iii) above) was dissolved in 10 mL of EtOH (95%). To this solution was added hydroxylamine hydrochloride (0.238 g, 3.42 mmol) and Et 3 N (0.48 mL, 3.44 mmol). After stirring at room temperature for 14 h, the solvent was removed and the residue was dissolved in EtOAc. The organic phase was washed with brine and H 2 O and was dried over Na 2 SO 4 . The crude product was purified by preparative RPLC with CH 3 CN:0.1 M NH 4 OAc as eluent, yielding the title compound as an amorphous powder (0.429 g, 72%) after freeze-drying.  
       1 H-NMR (400 MHz; CD 3 OD) rotamers: δ 7.35-7.1 (m, 5H), 6.90 (t, 1H, major rotamer), 6.85 (t, 1H, minor rotamer), 5.15 (s, 1H, major rotamer), 5.12 (m, 1H, minor rotamer), 5.08 (s, 1H, minor rotamer), 4.72 (m, 1H, major rotamer), 4.6-4.4 (m, 2H), 4.30 (m, 1H, major rotamer), 4.12 (m, 1H, major rotamer), 4.04 (m, 1H, minor rotamer), 3.94 (m, 1H, minor rotamer), 2.62 (m, 1H, minor rotamer), 2.48 (m, 1H, major rotamer), 2.22 (m, 1H, major rotamer), 2.10 (m, 1H, minor rotamer)  
       13 C-NMR (100 MHz; CD 3 OD): (carbonyl and amidine carbons, rotamers) δ 172.4, 171.9, 171.0, 152.3, 151.5  
      MS (m/z) 517.1, 519.0 (M−1) − , 519.1, 521.0 (M+1) +   
     Preparation of Compound J (Ph(3-Cl)(5-OCH 2 CHF 2 )—(R)CH(OH)C(O)-Aze-Pab(OH))  
     (i) Ph(3-Cl)(5-OCH 2 CHF 2 )—(R)CH(OH)C(O)-Aze-Pab(Z)  
      Boc-Aze-Pab(Z) (see international patent application WO 97/02284, 92 mg, 0.197 mmol) was dissolved in 10 mL of EtOAc saturated with HCl(g) and allowed to react for 10 min. The solvent was evaporated and the residue was mixed with Ph(3-Cl)(5-OCH 2 CHF 2 )—(R)CH(OH)C(O)OH (50 mg, 0.188 mmol; see Preparation C (v) above), PyBOP (109 mg, 0.209 mmol) and finally diisopropylethyl amine (96 mg, 0.75 mmol) in 2 mL of DMF. The mixture was stirred for 2 h and then poured into 50 mL of water and extracted three times with EtOAc. The combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The crude product was flash chromatographed on silica gel with EtOAc:MeOH (9:1). Yield: 100 mg (87%).  
       1 H NMR (300 MHz, CD 3 OD, mixture of rotamers) δ 7.85-7.75 (m, 2H), 7.45-7.25 (m, 7H), 7.11 (m, 1H, major rotamer), 7.08 (m, 1H, minor rotamer), 7.05-6.9 (m, 2H), 6.13 (bt, 1H), 5.25-5.05 (m, 3H), 4.77 (m, 1H, partially hidden by the CD 3 OH signal), 4.5-3.9 (m, 7H), 2.64 (m, 1H, minor rotamer), 2.47 (m, 1H, major rotamer), 2.25 (m, 1H, major rotamer), 2.13 (m, 1H, minor rotamer)  
     (ii) Ph(3-Cl)(5-OCH 2 CHF 2 )—(R)CH(OH)C(O)-Aze-Pab(OH)  
      Hydroxylamine hydrochloride (65 mg, 0.94 mmol) and triethylamine (0.319 g, 3.16 mmol) were mixed in 8 mL of THF and sonicated for 1 h at 40° C. Ph(3-Cl)(5-OCH 2 CHF 2 )—(R)CH(OH)C(O)-Aze-Pab(Z) (96 mg, 0.156 mmol; see step (i) above) was added with 8 mL more of THF. The mixture was stirred at 40° C. for 4.5 days. The solvent was evaporated and the crude product was purified by preparative RPLC with CH 3 CN:0.1M NH 4 OAc (40:60).  
      Yield: 30 mg (38%). Purity: 99%.  
       1 H NMR (300 MHz, CD 3 OD, mixture of rotamers) δ 7.6-7.55 (m, 2H), 7.35-7.3 (m, 2H), 7.12 (m, 1H, major rotamer), 7.09 (m, 1H, minor rotamer), 7.05-6.9 (m, 2H), 6.15 (triplet of multiplets, 1H), 5.15 (m, 1H, minor rotamer), 5.13 (s, 1H, major rotamer), 5.08 (s, 1H, minor rotamer), 4.77 (m, 1H, major rotamer), 4.5-4.2 (m, 5H), 4.08 (m, 1H, major rotamer), 3.97 (m, 1H, minor rotamer), 2.66 (m, 1H, minor rotamer), 2.50 (m, 1H major rotamer), 2.27 (m, 1H, major rotamer), 2.14 (m, 1H, minor rotamer).  
       13 C-NMR (100 MHz; CD 3 OD): (carbonyl and/or amidine carbons, mixture of rotamers) δ 172.8, 172.2, 171.4, 159.1, 158.9, 154.2.  
      APCI-MS: (M+1)=497/499 m/z  
     Methods 1 and 2: Preparation of Salts of Compound A  
     Method 1: General Method for Salt Preparation  
      The following generic method was employed to prepare salts of Compound A: 200 mg of Compound A (see Preparation A above) was dissolved in 5 mL of MeOH. To this solution was added a solution of the relevant acid (1.0 molar equivalent) dissolved in 5 mL of MeOH. After stirring for 10 minutes at room temperature, the solvent was removed by way of a rotary evaporator. The remaining solid material was re-dissolved in 8 mL of acetonitrile:H 2 O (1:1). Freeze-drying afforded colorless amorphous material in each case.  
      Acids Employed:  
     
         
          (1S)-(+)-10-camphorsulfonic  
          malic  
          cyclohexylsulphamic  
          phosphoric  
          dimethylphosphoric  
          p-toluenesulphonic  
          L-lysine  
          L-lysine hydrochloride  
          saccharinic  
          methanesulphonic  
          hydrochloric  
       
    
      Appropriate characterising data are shown in Table 1.  
                               TABLE 1                                       δ ppm (MeOD)                       H18, H19, H24           Mw   Mw       (see structure at end       Salt   acid   salt   LRMS   of Method 9 below)                                                    (1S)-(+)-10-   232.20   729.20   230.9   7.57, 7.68, 3.97       camphorsulfonate           495.1                   497.0                   727.3       maleate   116.07   612.97   114.8   7.45, 7.64, 3.89                   495.1                   497.0       cyclohexylsulphamate   179.24   676.14   177.9   7.44, 7.64, 3.89                   495.1                   496.9                   674.3                   676.1       phosphate   97.99   594.89   495.1   7.37, 7.61, 3.84                   497.0                   593.1       dimethylphosphate   126.05   622.95   124.9   7.50, 7.66, 3.92                   495.1                   497.0                   621.2                   623.0       p-toluenesulphonate   172.20   669.10   170.9   7.54, 7.71, 3.95                   495.1                   497.0       L-lysine   146.19   643.09   145.0   7.36, 7.60, 3.83                   495.1                   497.0       L-lysine hydrochloride   182.65   679.55   495.1   7.36, 7.60, 3.83                   497.0                   531.1                   (HCl)       saccharinate   183.19   680.09   181.9   7.44, 7.64, 3.89                   495.1                   497.0       methanesulphonate   96.11   593.01   495.1   7.57, 7.68, 3.97                   497.0                   591.2                   593.1       hydrochloride   36.46   533.36   495.1   7.55, 7.67, 3.95                   496.9                   531.1                   532.5                   535.2                  
 
      All salts formed in this Method were amorphous.  
     Method 2  
      Further amorphous salts of Compound A were made using analogous techniques to those described in Method 1 above from the following acids: 
      hydrobromic acid (1:1 salt)     hydrochloric acid (1:1 salt)     sulphuric acid (1:0.5 salt)     1,2-ethanedisulfonic acid (1:0.5 salt)     1S-camphorsulfonic acid (1:1 salt)     (+/−)-camphorsulfonic acid (1:1 salt)     ethanesulfonic acid (1:1 salt)     nitric acid (1:1 salt)     toluenesulfonic acid (1:1 salt)     methanesulfonic acid (1:1 salt)     p-xylenesulfonic acid (1:1 salt)     2-mesitylenesulfonic acid (1:1 salt)     1,5-naphthalenesulfonic acid (1:0.5 salt)     naphthalenesulfonic acid (1:1 salt)     benzenesulfonic acid (1:1 salt)     saccharinic acid (1:1 salt)     maleic acid (1:1 salt)     phosphoric acid (1:1 salt)     D-glutamic acid (1:1 salt)     L-glutamic acid (1:1 salt)     D,L-glutamic acid (1:1 salt)     L-arginine (1:1 salt)     L-lysine (1:1 salt)     L-lysine hydrochloride (1:1 salt)     glycine (1:1 salt)     salicylic acid (1:1 salt)     tartaric acid (1:1 salt)     fumaric acid (1:1 salt)     citric acid (1:1 salt)     L-(−)-malic acid (1:1 salt)     D,L-malic acid (1:1 salt)     D-gluconic acid (1:1 salt)    

     Method 3: Preparation of Amorphous Compound A, ethanesulfonic acid salt  
      Compound A (203 mg; see Preparation A above) was dissolved in ethanol (3 mL) and ethanesulfonic acid (1 eq., 95%, 35 μL) was added to the solution. The mixture was stirred for a few minutes, and then the solvent was evaporated. The resulting oil was slurried in iso-octane and evaporated to dryness until a solid material was obtained. Finally, the substance was re-slurried in iso-octane and the solvent evaporated again resulting in a white, dry, amorphous solid. The substance was vacuum dried at 40° C. overnight.  
     Methods 4 to 9: Preparation of Crystalline Compound A, ethanesulfonic acid salt  
     Method 4: Crystallisation of Amorphous Material  
      Amorphous Compound A, ethanesulfonic acid salt (17.8 mg; see Method 3 above) was slurried in methyl iso-butyl ketone (600 μL). After 1 week, crystalline needles were observed, which were filtered off and air-dried.  
     Methods 5 to 7: Reaction Crystallisations (without Anti-solvent)  
     Method 5  
      Compound A (277 mg; see Preparation A above) was dissolved in methyl iso-butyl ketone (3.1 mL). Ethanesulfonic acid was added (1 eq., 95%, 48 μL). Precipitation of amorphous ethanesulfonate salt occurred immediately. More methyl iso-butyl ketone (6 mL) was added and the slurry was treated with ultrasound. Finally, a third portion of methyl iso-butyl ketone (3.6 mL) was added and then the slurry was left overnight with stirring (magnetic stirrer). The next day, the substance had transformed into crystalline needles. The slurry was filtered off, washed with methyl iso-butyl ketone (0.5 mL) and air dried.  
     Method 6  
      Compound A (236 mg; see Preparation A above) was dissolved at room temperature in methyl iso-butyl ketone (7 mL). Ethanesulfonic acid (1 eq., 41 μL) was mixed with 2 mL of methyl iso-butyl ketone in a vial. The solution of Compound A was seeded with crystalline Compound A, ethanesulfonic acid salt (see Methods 4 and 5 above). Then, 250 μL of the methyl iso-butyl ketone solution of ethanesulfonic acid was added in portions over 45 minutes. The solution was seeded again, and the temperature was increased to 30° C. Then, 500 μL of the methyl iso-butyl ketone solution was added over approximately 1 hour. The resulting slurry was left overnight before a final amount of the methyl iso-butyl ketone/acid solution was added over 20 minutes. The vial was rinsed with 1.5 mL of methyl iso-butyl ketone, which was added to the slurry. After a further 6 hours, the crystals were filtered off, washed with methyl iso-butyl ketone (2 mL) and dried under reduced pressure at 40° C. A total of 258 mg of crystalline salt was obtained which corresponds to a yield of approximately 87%.  
     Method 7  
      Compound A (2.36 g; see Preparation A above) was dissolved in methyl iso-butyl ketone (90 mL). Seed crystals (10 mg) of Compound A, ethanesulfonic acid salt (see Methods 4 to 6 above) were added to the solution, and then ethanesulfonic acid (40 μL) was added in two portions. Further seed crystals (12 mg) and two portions of ethanesulfonic acid (2×20 μL) were then added. The slurry was diluted with methyl iso-butyl ketone (15 mL) before the addition of ethanesulfonic acid was continued. A total amount of 330 μL ethanesulfonic acid was added, in portions, over 1 hour. A small amount of seed crystals was added and, finally, the slurry was left overnight with stirring. The next day, the crystals were filtered off, washed with methyl iso-butyl ketone (2×6 mL) and dried under reduced pressure at 40° C. After drying, a total of 2.57 g of white, crystalline product was obtained corresponding to a yield of 89%.  
     Methods 8 and 9: Reaction Crystallizations (with Anti-Solvent)  
     Method 8  
      Compound A (163 mg; see Preparation A above) was dissolved in iso-propanol (1.2 mL). The solution was heated to 35° C. Ethanesulfonic acid was added (28 μL). Then, ethyl acetate (4.8 mL) was added and the solution was seeded with crystalline Compound A, ethanesulphonic acid salt (see Methods 4 to 7 above). Crystallization started almost immediately. The slurry was left for about 80 minutes at 35° C. before being allowed to cool to ambient temperature (21° C.). Two hours later, the crystals were filtered off, washed three times with ethyl acetate (3×0.4 mL), and dried under reduced pressure at 40° C. A total of 170 mg of crystalline title product was obtained which corresponds to a yield of approximately 82%.  
     Method 9  
      Compound A (20.0 g; see Preparation A above) was dissolved in iso-propanol (146.6 mL) at 40° C. and ethanesulfonic acid (3.46 mL, 95%, 1 eq.) was added to the solution. To the resulting clear solution, seed crystals of Compound A, ethanesulfonic acid salt were added (50 mg; see Methods 4 to 8 above). Then, ethyl acetate (234 mL) was added over 10 minutes. The resulting slightly opaque solution was seeded once more (70 mg) and left for one hour at 40° C. with stirring to allow for crystallization to start. After this, a total of 352 mL of ethyl acetate was added at a constant rate over one hour. When all of the ethyl acetate had been added, the slurry was left for 1 hour, before being cooled to 21° C. over 2 hours. The crystallization was allowed to continue for 1 hour at 21° C. before the crystals were filtered off, washed twice with ethyl acetate (50 mL+60 mL) and finally, dried under reduced pressure at 40° C. overnight. A total of 21.6 g of a white, crystalline salt was obtained, corresponding to a yield of approximately 90%.  
      Compound A, ethanesulfonic acid salt was characterised by NMR as follows: 23 mg of the salt was dissolved in deuterated methanol (0.7 mL) troscopy. A combination of 1D ( 1 H,  13 C and selective NOE) and 2D (gCOSY, gHSQC and gHMBC) NMR experiments were used. All data were in good agreement with the theoretical structure of the salt, shown below. The molecule exists in two conformations in methanol. Based on the integral of the peak assigned to H5 (dominant conformer) and peak assigned to H5′ (other conformer), the ratio between the two conformers was found to be 70:30. H22 could not be observed as these protons were in fast exchange with the solvent CD 3 OD.  
                 
 
      Both the proton and the carbon resonance corresponding to position 1 are split due to the spin-coupling with the two fluorine nuclei in that position. The coupling constants are  2 J HF =73 Hz and  1 J CF =263 Hz.  
       1 H and  13 C NMR chemical shift assignment and proton-proton correlations are shown in Table 2.  
                               TABLE 2                       Atom         13 C shift/     1 H shift/ppm b  and           No.   Type   ppm a     multiplicity c     J HH /Hz                                                     1   CH   117.5 e     6.90 (t)   73 ( 2 J HF )            1′       117.5 e     6.88 (t)        2   C   153.5            2′       153.5        3   CH   120.0   7.15 (s)            3′       119.7   7.13 (s)        4   C   136.2            4′       135.9        5   CH   125.0   7.36 (s)            5′       124.9   7.31 (s)        6   C   144.5            6′       145.3        7   CH   117.3   7.20 (s)            7′       117.2   7.15 (s)        8   CH   72.0   5.20 (s)            8′       74.0   5.12 (s)        9   CO   173.1            9′       173.8       11   CH 2     51.6   a: 4.38 (m)                   b: 4.21 (m)           11′       49.0   a: 4.06 (m)                   b: 3.99 (m)       12   CH 2     21.7   a: 2.55 (m)                   b: 2.29 (m)           12′       23.2   a: 2.70 (m)                   b: 2.15 (m)       13   CH   63.1   4.80 (m)           13′       66.2   5.22 (m)       14   CO   172.9           14′       173.6       15   NH       8.76 (t, br)   5.2           15′           8.79 (t, br)   5.2       16   CH 2     43.5   4.59 (AB-pattern)   15.9                   4.46 (AB-pattern)   15.9           16′       43.6   4.53 (AB-pattern)   15.9                   4.49 (AB-pattern)   15.9       17   C   146.9           17′       147.0       18   CH   129.1   7.56 (d)   7.8           18′       129.1   7.57 (d)   7.8       19   CH   129.2   7.67 (d)   7.8           19′       129.4   7.70 (d)   7.8       20   C   124.9   —           20′       124.9       21   C   162.4           21′       162.3       22   NH 2         Not observed       24   CH 3     64.8   3.96 (s)       101    CH3       1.28 (t)   7.4       102    CH2       2.77 (m)   7.4                   a Relative to the solvent resonance at 49.0 ppm.              b Relative to the solvent resonance at 3.30 ppm.              c s = singlet, t = triplet, m = multiplet, br = broad, d = doublet              d Obtained in the gCOSY experiment.              e The resonance is a triplet due to coupling with the two fluorine nuclei.              1 J CF  = 263 Hz.               
      HRMS calculated for C 24 H 29 ClF 2 N 4 O 8 S (M−H)hu − 605.1284, found 605.1296.  
      Crystals of Compound A, ethanesulfonic acid salt (obtained by way of one or more of Examples 4 to 9 above) were analyzed by XPAD and the results are tabulated below (Table 3) and are shown in FIG. 1.  
                       TABLE 3                       d value (Å)   Intensity (%)   Intensity                                            16.5   10   m       12.2   74   vs       11.0   4   w       9.0   33   s       8.3   3   vw       7.6   6   w       6.4   4   w       6.2   12   m       6.0   7   m       5.9   10   m       5.5   15   m       5.4   100   vs       5.1   7   m       4.66   29   s       4.60   36   s       4.31   57   s       4.25   18   m       4.19   20   m       4.13   12   m       4.00   12   m       3.87   13   m       3.83   6   w       3.76   7   m       3.72   6   w       3.57   9   m       3.51   7   m       3.47   5   w       3.39   3   vw       3.31   11   m       3.26   10   m       3.21   8   m       3.16   4   w       3.03   8   m       2.78   4   w       2.74   5   w       2.67   3   vw       2.56   5   w       2.50   5   w       2.46   7   m       2.34   4   w       2.21   5   w       2.00   3   vw       1.98   3   vw                  
 
      DSC showed an endotherm with an extrapolated melting onset temperature of ca. 131° C. TGA showed a decrease in mass of ca. 0.2% (w/w) around the melting point. DSC analysis repeated with a sample of lower solvent content showed a melting onset temperature of ca. 144° C.  
     Method 10: Preparation of Amorphous Compound A, benzenesulfonic acid salt  
      Compound A (199 mg; see Preparation A above) was dissolved in ethanol (2 mL). Benzenesulfonic acid (1 eq. 90%, 70 mg) was dissolved in ethanol (1 mL) in a vial. The ethanol solution of the acid was added to the solution of Compound A and the vial was rinsed with 1 mL ethanol, which was then added to the mixture. The mixture was stirred for a few minutes, and then the ethanol was evaporated until an oil was formed. Ethyl acetate (3 mL) was added and the solvent was evaporated again to dryness. An amorphous solid was formed.  
     Methods 11 to 13: Preparation of Crystalline Compound A, benzenesulfonic acid salt  
     Method 11: Crystallisation of Amorphous Material  
      Amorphous Compound A benzenesulfonic acid salt (20.7 mg; see Method 10 above) was slurried in ethyl acetate (600 TL). After 5 days, crystalline needles were observed in the slurry.  
     Methods 12 and 13: Reaction Crystallisations  
     Method 12  
      Compound A (128 mg; see Preparation A above) was dissolved in ethyl acetate (3 mL). The solution was seeded with the slurry from Method 11 above. Then, benzenesulfonic acid was added (1 eq., 90%, 45 mg). Precipitation of benzenesulphonic acid salt occurred immediately. iso-Propanol was added to the slurry (0.8 mL) and the mixture was seeded again. Two days later, the substance had transformed into crystalline needles. The slurry was filtered off, washed with ethyl acetate (3×0.2 mL) and dried for a short time under vacuum at 40° C. A total of approximately 140 mg of white solid was obtained.  
     Method 13  
      Compound A (246 mg; see Preparation A above) was dissolved in iso-propanol (1.52 mL). Benzenesulfonic acid was added (88 mg, 90%). To the clear solution, ethyl acetate was added (3 mL), and then the mixture was seeded to initiate crystallisation. After 1 hour, more ethyl acetate was added (2.77 mL). Finally, the slurry was allowed to crystallise overnight before the crystals were filtered off, washed with ethyl acetate (3×0.3 mL) and dried at 40° C. under vacuum. A total of 279 mg salt was obtained which corresponds to a yield of approximately 86%.  
      Compound A, benzenesulfonic acid salt was characterised by NMR as follows: 20 mg of the salt was dissolved in deuterated methanol (0.7 mL). A combination of 1D ( 1 H,  13 C and selective NOE) and 2D (gCOSY, gHSQC and gHMBC) NMR experiments were used. All data were in good agreement with the theoretical structure of the salt, shown below. The molecule exists in two conformations in methanol. Based on the integral of the peak assigned to H12 (dominant conformer) and peak assigned to H12′ (other conformer), the ratio between the two conformers was found to be 70:30. H22 could not be observed as these protons were in fast exchange with the solvent CD 3 OD.  
                 
 
      Both the proton and the carbon resonance corresponding to position 1 are split due to the spin-coupling with the two fluorine nuclei in that position. The coupling constants are  2 J HF =74 Hz and  1 J CF =260 Hz.  
       1 H and  13 C NMR chemical shift assignment and proton-proton correlations are shown in Table 4.  
                               TABLE 4                       Atom             1 H shift/ppm b  and           No.   Type     13 C shift/ppm a     multiplicity c     J HH /Hz                                                     1   CH   117.5 e     6.89 (t)   74 ( 2 J HF )            1′       117.5 e     6.87 (t)        2   C   153.5            2′       153.5        3   CH   120.1   7.15 (s)            3′       119.7   7.12 (s)        4   C   136.2            4′       135.9        5   CH   125.1   7.35 (s)            5′       124.9   7.31 (s)        6   C   144.5            6′       145.3        7   CH   117.3   7.20 (s)            7′       117.2   7.14 (s)        8   CH   72.8   5.20 (s)            8′       74.0   5.12 (s)        9   CO   173.1            9′       173.8       11   CH 2     51.6   a: 4.37 (m)                   b: 4.20 (m)           11′       49.0   a: 4.05 (m)                   b: 3.98 (m)       12   CH 2     21.7   a: 2.53 (m)                   b: 2.28 (m)           12′       23.2   a: 2.69 (m)                   b: 2.14 (m)       13   CH   63.1   4.79 (m)           13′       66.2   5.22 (m)       14   CO   172.9           14′       173.6       15   NH       8.75 (t, br)   5.3           15′           8.78 (t, br)   5.3       16   CH 2     43.5   4.59 (AB-pattern)   16.0 and 5.2                   4.44 (AB-pattern)   16.0 and 4.8           16′       43.6   4.51 (AB-pattern)   16.0                    4.46 (AB-pattern)   16.0        17   C   146.9           17′       147.0       18   CH   129.2   7.54 (d)   8.3           18′       129.2   7.56 (d)   8.3       19   CH   129.3   7.66 (d)   8.3           19′       129.4   7.69 (d)   8.3       20   C   124.9   —           20′       124.9       21   C   162.4           21′       162.4       22   NH 2         Not observed       24   CH 3     64.8   3.95 (s)       101    CH   126.9   7.81 (m)       102    CH   129.1   7.41 (m)       103    CH   131.2   7.42 (m)       104    C   146.4                   a Relative to the solvent resonance at 49.0 ppm.              b Relative to the solvent resonance at 3.30 ppm.              c s = singlet, t = triplet, m = multiplet, br = broad, d = doublet.              d Obtained in the gCOSY experiment.              e The resonance is a triplet due to coupling with the two fluorine nuclei.              1 J CF  = 260 Hz.              f connectivity difficult to determine due to overlap between resonance 102 and 103               
      HRMS calculated for C 28 H 29 ClF 2 N 4 O 8 S (M−H) −  653.1284, found 653.1312.  
      Crystals of Compound A, benzenesulfonic acid salt (obtained by way of one or more of Examples 11 to 13 above) were analyzed by XRPD and the results are tabulated below (Table 5) and are shown in FIG. 2.  
                       TABLE 5                       d value (Å)   Intensity (%)   Intensity                                            14.2   12   m       12.6   55   s       10.2   49   s       7.5   8   m       6.4   5   w       6.3   30   s       6.1   5   w       5.9   100   vs       5.7   20   m       5.4   9   m       5.3   11   m       5.1   10   m       4.96   3   vw       4.83   27   s       4.73   72   vs       4.54   23   s       4.50   10   m       4.35   28   s       4.30   38   s       4.24   24   s       4.17   28   s       4.09   60   vs       4.08   61   vs       3.96   29   s       3.91   15   m       3.77   22   s       3.62   11   m       3.52   20   m       3.31   44   s       3.19   8   m       3.15   11   m       3.09   8   m       3.00   7   m       2.89   3   vw       2.86   4   w       2.79   7   m       2.76   6   w       2.72   5   w       2.59   6   w       2.56   9   m       2.54   9   m       2.49   7   m       2.38   8   m       2.16   4   w       2.03   3   vw                  
 
      DSC showed an endotherm with an extrapolated melting onset temperature of ca. 152° C. TGA showed a decrease in mass of ca. 0.1% (w/w) around the melting point.  
     Method 14: Preparation of Amorphous Compound A, n-propanesulfonic acid salt  
      Compound A (186 mg; see Preparation A above) was dissolved in iso-propanol (1.39 mL) and n-propanesulfonic acid (1 eq., 95%, 39 TL) was added. Ethyl acetate (5.6 mL) was added and the solvent was evaporated until a dry, amorphous solid was formed.  
     Methods 15 and 16: Preparation of Crystalline Compound A, n-propanesulfonic acid salt  
     Method 15: Crystallisation of Amorphous Material  
      Amorphous Compound A, n-propanesulfonic acid salt (20 mg; see Method 14 above) was dissolved in iso-propanol (60 TL) and iso-propyl acetate (180 TL) was added. After three days crystalline needles were observed.  
     Method 16: Reaction Crystallisation  
      Compound A (229 mg; see Preparation A above) was dissolved in iso-propanol (1.43 mL). n-Propanesulfonic acid was added (1 eq., 95%, 48 TL). Ethyl acetate was added (2 mL), and then the solution was seeded with crystalline salt from Method 15 above. Further ethyl acetate was added (5 mL) and the slurry was left overnight to crystallize. The crystals were filtered off, washed with ethyl acetate (3×0.3 mL) and dried under vacuum at 40° C.  
      Compound A, n-propanesulfonic acid salt was characterised by NMR as follows: 13 mg of the salt was dissolved in deuterated methanol (0.7 mL) troscopy. A combination of 1D ( 1 H,  13 C) and 2D (gCOSY) NMR experiments were used. All data were in good agreement with the theoretical structure of the salt, shown below. The molecule exists in two conformations in methanol. Based on the integral of the peak assigned to H12 (dominant conformer) and peak assigned to H12′ (other conformer), the ratio between the two conformers was found to be 65:35. H22 could not be observed as these protons were in fast exchange with the solvent  
                 
 
      Both the proton and the carbon resonance corresponding to position 1 are split due to the spin-coupling with the two fluorine nuclei in that position. The coupling constants are  2 J HF =74 Hz and  1 J CF =260 HZ.  
       1 H and  13 C NMR chemical shift assignment and proton-proton correlations are shown in Table 6.  
                               TABLE 6                       Atom             1 H shift/ppm b  and           No.   Type     13 C shift/ppm a     multiplicity c     J HH /Hz                                                     1   CH   117.5 e     6.89 (t)   74 ( 2 J HF )            1′       117.5 e     6.88 (t)        2   C   153.5            2′       153.5        3   CH   120.0   7.16 (s)            3′       119.7   7.13 (s)        4   C   136.2            4′       135.9        5   CH   125.1   7.36 (s)            5′       124.9   7.31 (s)        6   C   144.5            6′       145.3        7   CH   117.3   7.20 (s)            7′       117.2   7.16 (s)        8   CH   72.9   5.20 (s)            8′       74.1   5.12 (s)        9   CO   173.1            9′       173.8       11   CH 2     51.6   a: 4.37 (m)                   b: 4.20 (m)           11′       49.0   a: 4.06 (m)                   b: 3.98 (m)       12   CH 2     21.7   a: 2.53 (m)                   b: 2.29 (m)           12′       23.2   a: 2.69 (m)                   b: 2.15 (m)       13   CH   63.1   4.80 (m)           13′       66.2   5.22 (m)       14   CO   172.9           14′       173.8       15   NH       8.75 (t, br)   5.5           15′           8.79 (t, br)   5.5       16   CH 2     43.5   4.59 (AB-pattern)   16.0 and 6.6                   4.45 (AB-pattern)   16.0 and 5.3           16′       43.6   4.51                   4.50       17   C   146.9           17′       147.0       18   CH   129.1   7.54 (d)   8.5           18′       129.2   7.57 (d)   8.5       19   CH   129.2   7.67 (d)   8.5           19′       129.4   7.69 (d)   8.5       20   C   124.9   —           20′       124.9       21   C   162.4           21′       162.4       22   NH 2         Not observed       24   CH 3     64.7   3.96 (s)       101    CH   13.7   1.0 (t)       102    CH   19.6   1.78 (m)       103    CH   54.6   2.75 (m)                   a Relative to the solvent resonance at 49.0 ppm.              b Relative to the solvent resonance at 3.30 ppm.              c s = singlet, t = triplet, m = multiplet, br = broad, d = doublet.              d Obtained in the gCOSY experiment.              e The resonance is a triplet due to coupling with the two fluorine nuclei.              1 J CF  = 260 Hz.               
      HRMS calculated for C 25 H 31 ClF 2 N 4 O 8 S (M−H) −  619.1441, found 619.1436.  
      Crystals of Compound A, n-propanesulfonic acid salt (obtained by way of one or more of Examples 15 and 16 above) were analyzed by XRPD and the results are tabulated below (Table 7) and are shown in FIG. 3.  
                       TABLE 7                       d value (Å)   Intensity (%)   Intensity                                            14.0   4   w       12.4   87   vs       10.0   30   s       8.0   3   vw       7.5   7   m       7.0   0.6   vw       6.7   1   vw       6.4   1   vw       6.2   12   m       6.1   3   vw       5.8   100   vs       5.7   11   m       5.5   3   vw       5.4   5   w       5.3   5   w       5.2   2   vw       5.1   3   vw       4.94   3   vw       4.78   21   s       4.68   42   s       4.51   10   m       4.49   7   m       4.40   5   w       4.32   10   m       4.29   10   m       4.25   22   s       4.19   14   m       4.14   15   m       4.07   23   s       4.04   20   m       3.94   16   m       3.88   10   m       3.73   15   m       3.65   2   vw       3.59   3   vw       3.48   18   m       3.28   23   m       3.12   4   w       3.06   3   vw       2.97   6   w       2.84   2   vw       2.81   3   vw       2.76   2   vw       2.73   3   vw       2.70   2   vw       2.57   2   vw       2.54   6   w       2.51   6   w       2.46   8   m       2.42   2   vw       2.39   3   vw       2.36   3   vw       2.32   2   vw       2.14   3   vw       2.01   2   vw                  
 
      DSC showed an endotherm with an extrapolated melting onset temperature of ca. 135° C. TGA showed no decrease in mass around the melting point.  
     Method 17  
     Method 17-A: Preparation of amorphous Compound A n-butane sulfonic acid salt  
      Amorphous Compound A (277 mg) was dissolved in IPA (1.77 ml) and butane sulfonic acid (approx. 1 eq. 70 μL) was added. Ethyl acetate (6 ml) was added and the solvent was evaporated until dry, amorphous solid was formed.  
     Method 17-B: Preparation of Crystalline Compound A Butane sulfonic acid salt  
      Amorphous Compound A butane sulfonic acid salt (71.5 mg; see preparation above) was slurried in ethyl acetate (500 μl) over night. The crystals were filtered off and were air-dried.  
      Compound A, butanesulfonic acid salt was charaterised by NMR as follows:  
      21.6 mg of the salt was dissolved in deuterated dimethylsulfoxide (0.7 ml) and was investigated with  1 H and  13 C NMR spectroscopy.  
      The spectra are very similar to other salts of the same compound and in good agreement with the structure shown below. Most resonances in the spectra are present as sets of two peaks due to the slow rotation around the C9-N10 bond, which results in two atropisomers that simultaneously exist in the solution. This is shown for other salts of the same compound.  
                 
 
      The two fluorine nuclei in position 1 give rise to split resonances for the proton and the carbon in that position. The coupling constants are  2 J HF =73 Hz and  1 J CF =258 Hz. Chemical shifts for protons and carbons are presented in Table 1. Protons in position 22 and 24 are not detected due to chemical exchange. There is a very broad hump between 8 and 9 ppm in the proton spectrum corresponding to these protons.  
               TABLE 8                            1 H and  13 C NMR chemical shift assignment of Compound A       n-butanesulfonate salt in deuterated dimethylsulfoxide at 25° C.                                 Atom         13 C shift/     1 H shift/ppm b  and           No.   Typ   ppm a     multiplicity c     J HH /Hz                                          1   CHF 2     116.3 d     7.29 (t)   73 ( 2 J HF )            1′       116.3 d     7.28 (t)   73 ( 2 J HF )        2   C   151.5   na   na            2′       151.3   na   na        3   CH   118.0   7.25 (t) e     nd            3′       117.6   7.21 (t) e     nd        4   C   133.8   na   na            4′       133.4   na   na        5   CH   123.8   7.34 (t) e     nd            5′       123.6   7.25 (t) e     nd        6   C   144.5   na   na            6′       145.2   na   na        7   CH   116.3   7.19 (t) e     nd            7′       116.1   7.12 (t) e     nd        8   CH   70.9   5.13 (s)   na            8′       71.2   4.99 (s)   na        9   CO   170.6   na   na            9′       171.1   na   na       11   CH 2     50.0   a: 4.24 (m) b: 4.12 (m)   nd           11′       46.9   3.85 (m)   nd       12   CH 2     20.5   a: 2.41 (m) b: 2.10 (m)   nd           12′       21.7   a: 2.60 (m) b: 2.02 (m)   nd       13   CH   61.2   4.65 (dd)   5.6 and 8.9           13′       63.9   5.12 (m)   nd       14   CO   170.2   na   na           14′       171.0   na   na       16   CH 2     41.8   4.38 (m)   nd           16′       42.0   4.38 (m)   nd       17   C   144.7   na   na       18   CH   127.5   7.44 (d)   8.2               127.6   7.44   nd       19   CH   127.8   7.66 (d)   8.2       20   C   125.1   na   na       21   C   157.9   na   na       24   CH 3     63.3   3.83 (s)   na           24′       63.3   3.82 (s)   na       26   CH 2     51.4   2.41 (m)   nd       27   CH 2     27.3   1.52 (m)   nd       28   CH 2     21.7   1.30 (m)   nd       29   CH 3     14.0   0.83 (t)   7.3                   a Relative to the solvent resonance at 49.0 ppm.              b Relative to the solvent resonance at 3.30 ppm.              c s = singlet, d = doublet, dd = doublet of doublets, t = triplet, m = multiplet.              d The resonance is a triplet due to coupling with the two fluorine nuclei F1.              1 J CF  = 258 Hz.              e The  4 J HH  coupling with the meta-protons is not fully resolved.            na = not applicable,            nd = not determined             
 
      HRMS calculated for C 26 H 32 ClF 2 N 4 O 8 S (M−H) −  633.1597, found 633.1600  
      Crystals of Compound A n-butanesulfonic acid salt (obtained as described above in Method 17-B) were analyzed by XRPD and the results are tabulated below (Table 9) and are shown in FIG. 4.  
                       TABLE 9                       d-value (Å)   Intensity (%)   Intensity                                            14.3   8   m       12.8   81   vs       10.3   44   s       8.2   4   w       7.7   13   m       6.7   2   vw       6.4   8   m       6.2   18   m       6.0   100   vs       5.8   29   s       5.6   4   w       5.4   11   m       5.3   16   m       5.1   15   m       4.98   6.5   w       4.91   34   s       4.76   56   s       4.57   20   m       4.42   13   m       4.36   19   m       4.30   45   s       4.18   42   s       4.13   88   vs       4.01   34   s       3.92   28   s       3.82   18   m       3.64   6.6   w       3.58   16   m       3.47   5   w       3.44   6   w       3.38   12   m       3.35   32   s       3.32   22   s       3.29   12   m       3.20   8   m       3.17   9   m       3.02   12   m       2.90   6   w       2.81   3.9   vw       2.75   3   vw       2.64   3.5   vw       2.59   10   m       2.57   8   m       2.50   4   w       2.45   5   w       2.40   6   w       2.31   3   vw                  
 
      DSC showed an endotherm with an extrapolated melting onset temperature of ca 118° C. and TGA showed a 0.04% weight loss.  
     Method 18: Preparation of Salts of Compound B  
     Method 18-A: General Method for Salt Preparation  
      The following generic method was employed to prepare salts of Compound B: 200 mg of compound B (see Preparation B above) was dissolved in 5 mL of MIBK (methyl isobutyl ketone). To this solution was added a solution of the relevant acid (1.0 or 0.5 molar equivalent, as indicated in Table 10) dissolved in 1.0 mL of MIBK. After stirring for 10 minutes at room temperature, the solvent was removed by way of a rotary evaporator. The remaining solid material was re-dissolved in about 8 mL of acetonitrile:H 2 O (1:1). Freeze-drying afforded colorless amorphous material in each case.  
      Acid Employed:  
     
         
          Esylate (ethanesulfonic acid)  
          Besylate (benzene sulfonic acid)  
          Cyclohexylsulphamate  
          Sulphate  
          Bromide  
          p-Toluenesulphonate  
          2-Naphtalenesulfonate  
          Hemisulfate  
          Methanesulphonate  
          Nitrate  
          Hydrochloride  
       
    
      Appropriate characterising data are shown in Table 10 
                                   TABLE 10                                   Salt   Mw acid   Mw salt   MS ES-                                                            Esylate   110.13   643.01   108.8                       531.1                       641.0           Besylate   158.18   691.06   156.8                       531.1                       689.2           Cyclohexylsulphamate   179.24   712.12   177.9                       531.2                       710.4           Sulphate   98.08   630.96   531.1           Bromide   80.91   613.79   531.2                       613.1           p-Toluenesulphonate   172.20   705.08   170.9                       531.1                       703.1           2-   208.24   741.12   206.9           Naphtalenesulfonate           531.1                       739.3           Hemisulfate   98.07   1163.8    531.1                   (1:2)   631.0                   630.85                   (1:1)           Methanesulphonate   96.11   628.99   531.1                       627.1           Nitrate   63.01   595.89   531.0                       594.0           Hydrochloride   36.46   569.34   531.0                       569.0                      
 
      All salts formed in this Example were amorphous.  
     Method 18-B  
      Further amorphous salts of Compound B were made using analogous techniques to those described in Method 18-A above for the following acids: 
      1,2-Ethanedisulfonic (0.5 salt)     1S-Camphorsulfonic     (+/−)-Camphorsulfonic     p-Xylenesulfonic     2-Mesitylenesulfonic     Saccharin     Maleic     Phosphoric     D-glutamic     L-arginine     L-lysine     L-lysine*HCl    

     Method 18-C: Preparation of Amorphous Compound B, Hemi-1,5-naphtalenedisulfonic acid salt  
      Amorphous Compound B (110.9 mg) was dissolved in 2.5 mL 2-propanol and 0.5 equivalent of 1,5-naphthalene-disulfonic acid tetrahydrate was added (dissolved in 1 mL 2-propanol). The sample was stirred overnight. Only small particles (amorphous) or oil drops were observed by microscopy. The sample was evaporated to dryness.  
     Method 18-D: Preparation of Crystalline Compound B, hemi-1,5-naphtalenedisulfonic acid salt  
      The crystallization experiment was carried out at ambient temperature. Amorphous Compound B (0.4 gram) was dissolved in ethanol (1.5 mL) and 0.5 eq of 1,5-naphthalene-disulfonic acid tetrahydrate (1.35 gram, 10% in ethanol) was added. Heptane (0.7 mL) was then added until the solution became slightly cloudy. After about 15 minutes the solution became turbid. After about 30 minutes thin slurry was obtained and additional heptane (1.3 mL) was added. The slurry was than left overnight for ripening. To dilute the thick slurry, a mixture of ethanol and heptane (1.5 mL and 1.0 mL respectively) was added. After about 1 hour the slurry was filtered and the crystals were washed with a mixture of ethanol and heptane (1.5:1) and finally with pure heptane. The crystals were dried at ambient temperature in 1 day. The dry crystals weighed 0.395 g.  
     Method 18-E: Preparation of Crystalline Compound B, hemi-1,5-naphtalenedisulfonic acid salt  
      Amorphous Compound B (1.009 gr) was dissolved in 20 mL 2-propanol+20 mL ethyl acetate. 351.7 mg 1,5-naphtalene-disulfonic acid tetrahydrate, dissolved in 20 mL 2-propanol, was added drop by drop. Precipitation occurred in about 5 minutes. The slurry was stirred over night and then filtered.  
     Method 18-F: Preparation of Crystalline Compound B, hemi-1,5-naphtalenedisulfonic acid salt  
      430.7 mg of the 1,5-naphtalene-disulfonic acid salt was dissolved in 30 mL 1-propanol. The solution was heated to boiling in order to dissolve the substance. The solution was left over night at ambient temperature for crystallization and then the crystals were filtered off.  
     Method 18-G: Preparation of Crystalline Compound B, hemi-1,5-naphtalenedisulfonic acid salt  
      The mother liquid from Method 18-F was evaporated and the solid rest (61.2 mg) was dissolved in 6 mL acetonitrile/1-propanol, ratio 2:1. The solution was left overnight at ambient temperature to crystallize and then the crystals were filtered off.  
     Method 18-H: Preparation of Crystalline Compound B, hemi-1,5-naphtalenedisulfonic acid salt  
      The sample from Method 18-C was dissolved in about 2 mL methanol. Ethanol (about 3 mL) was added as anti-solvent at ambient temperature and seeds were added. No crystallization occurred, so solvents were evaporated (about half of the amount) and a new portion of ethanol (about 2 mL) and seeds were added. Crystalline particles were formed when stirred at ambient temperature during night.  
     Method 18-I: Preparation of Crystalline Compound B, hemi-1,5-naphtalenedisulfonic acid salt  
      Amorphous Compound B (104.1 mg) was dissolved in 2-propanol and 1 equivalent of 1,5-naphthalene-disulfonic acid tetrahydrate, dissolved in 2-propanol, was added In total, the 2-propanol amount was about 2.5 mL. The solution was stirred at 44° C. for about 80 minutes and a precipitate was formed. The particles were crystalline according to polarised light microscopy. The sample was filtered.  
     Method 18-J: Preparation of Crystalline Compound B, hemi-1,5-naphtalenedisulfonic acid salt  
      Compound B, hemi-1,5-naphtalenedisulfonic acid salt (56.4 mg) was dissolved in 1.5 mL methanol. Methyl ethyl ketone (3 mL) was added. Seeds were added to the solution and crystallization started. The crystals were filtered off, washed with methyl ethyl ketone and air dried.  
     Method 18-K: Preparation of crystalline Compound B, hemi-1,5-naphtalenedisulfonic acid salt  
      Amorphous Compound B (161,0 mg) was dissolved in 3.5 mL 1-Butanol and the solution was heated to 40° C. In another beaker 57.4 mg of naphthalene-disulfonic acid tetrahydrate was dissolved in 3 mL 1-Butanol. A couple of drops of the acid solution were added to the solution of compound B. Then seeds were added to the solution and after 2 hours the rest of the acid solution was added (at 40° C.) slowly. Then the temperature was slowly decreased to room temperature and the experiment was left under stirring overnight. The slurry was filtered, washed with 1-Butanol and dried under vacuum at 44° C. for 2 hours. The yield was 83%.  
      Characterisation  
      Crystals of Compound B, hemi-1,5-naphtalenedisulfonic acid salt, obtained by way of Method 18-D above, was charaterised by NMR as follows:  
      21.3 mg of the salt was dissolved in deuterated methanol, 0.7 ml was investigated with NMR spectroscopy. A combination of 1D ( 1 H,  13 C and selective NOE) and 2D (gCOSY, GHSQC and gHMBC) NMR experiments was used.  
      All data are in good agreement with the proposed structure, shown below. All carbons and the protons attached to carbons are assigned. Protons attached to heteroatoms are exchanged for deuterium from the solvent and are not detected. Most resonances in the 1D  1 H and  13 C NMR spectra are present as sets of two peaks. The reason for this is a slow rotation around the C9-N10 bond, which results in two atropisomers that simultaneously exist in the solution. The 1D NOE experiment is an evidence for this. When a resonance of one atropisomer is irradiated, the saturation is transferred to the corresponding peak of the other atropisomer. The resonances corresponding to the 1,5-naphtalenedisulfonate counter ion do not show atropisomerism.  
                 
 
      There are four fluorine atoms in the molecule. They give rise to split resonances for some protons and carbons. Both the proton and the carbon resonance corresponding to position 1 are split due to the spincoupling with the two fluorine nuclei in that position. The coupling constants are  2 J HF =73 Hz and  1 J CF =263 Hz. Further, the proton resonance corresponding to H19 is a distorted doublet with  3 J HF =6.9 Hz due to the spincoupling with the fluorine nuclei in position 18. Carbon resonances corresponding to C17, C18, C19 and C20 also exhibit couplings with these fluorine nuclei. The C17 and C20 resonances are triplets with  2 J CF =19 Hz and  3 J CF =11 Hz, respectively. The C18 resonance is a doublet of doublets with coupling constants  1 J CF =251 Hz and  3 J CF =8 Hz. The C19 resonance is a multiplet.  
      Comparing the magnitudes of integrals for resonances corresponding to the 1,5-naphtalenedisulfonate counter ion and the mother compound gives the stoichiometric relation of a single 1,5-naphtalenedisulfonate counter ion crystallized with two molecules of the mother compound.  
       1 H and  13 C NMR chemical shift assignment and proton-proton correlations are shown in Table 11.  
                                   TABLE 11                                 13 C           Through-bond       Atom       shift/     1 H shift/ppm b  and       correlation       No.   Typ   ppm a     multiplicity c     J HH /Hz   to  1 H d                                                           1   CHF 2     117.5 e     6.91 (t)   73 ( 2 J HF )   nd        1′       117.5 e     6.87 (t)   73 ( 2 J HF )   nd        2   C   153.5   na   na   na        2′       153.3   na   na   na        3   CH   120.0   7.14 (t) n     nd   5, 7        3′       119.6   7.11 (t) n     nd   5′, 7′        4   C   136.1   na   na   na        4′       135.8   na   na   na        5   CH   125.0   7.31 (t) n     nd   3, 7        5′       124.9   7.28 (t) n     nd   3′, 7′        6   C   144.4   na   na   na        6′       145.3   na   na   na        7   CH   117.2   7.16 (t) n     nd   3, 5        7′       117.1   7.12 (t) n     nd   3′, 5′        8   CH   72.9   5.15 (s)   na   nd        8′       73.6   5.07 (s)   na   nd        9   CO   173.0   na   na   na        9′       173.5   na   na   na       11   CH 2     51.5   a: 4.29 (m) b: 4.13   nd   12, 13       11′       48.6   (m)   nd   12′, 13′                   a: 4.01 (m) b: 3.93                   (m)       12   CH 2     21.7   a: 2.46 (m) b: 2.17   nd   11, 13       12′       22.8   (m)   nd   11′, 13′                   a: 2.61 (m) b: 2.03                   (m)       13   CH   62.8   4.70 (dd)   6.0 and   12       13′       65.8   5.14 (dd)   9.4   12′                       5.6 and                       9.1       14   CO   172.4   na   na   na       14′       173.2   na   na   na       16   CH 2     32.3   4.51 (m)   nd   nd       16′       32.5   4.51 (m)   nd   nd       17   C   121.0 f     na   na   na       18   CF   162.8 g     na   na   na       19   CH   112.7 i     7.35 (d)   6.9 ( 3 J HF )   nd       20   C   127.9 k     na   na   na       21   C   160.0   na   na   na       21′       159.9   na   na   na       24   CH 3     64.8   3.93 (s)   na   nd       24′       64.8   3.92 (s)   na   nd       25   C   142.4   na   na   na       26   CH   126.8   8.16 (d)   7.2   27, 28       27   CH   125.9   7.54 (dd)   8.6 and   26, 28                       7.2       28   CH   131.0   8.97 (d)   8.6   26, 27       29   C   131.1   na   na   na                   a Relative to the solvent resonance at 49.0 ppm.              b Relative to the solvent resonance at 3.30 ppm.              c s = singlet, d = doublet, dd = doublet of doublets, t = triplet, m = multiplet.              d Obtained in the gCOSY experiment.              e The resonance is a triplet due to coupling with the two fluorine nuclei F1.              1 J CF  = 263 Hz.              f The resonance is a triplet due to coupling to the two fluorine nuclei F18.              2 J CF  = 19 Hz.              g The resonance is a doublet of doublets due to coupling to the two fluorine nuclei F18.              1 J CF  = 251 Hz and              3 J CF  = 8 Hz.              i The resonance is a multiplet due to coupling to the two fluorine nuclei F18.              k The resonance is a triplet due to coupling to the two fluorine nuclei F18.              3 J CF  = 11 Hz.              n The  4 J HH  coupling with the meta-protons is not fully resolved.            na = not applicable,            nd = not determined               
      Crystals of Compound B, hemi-1,5-naphtalenedisulfonic acid salt (obtained by way of Method 18-I above, were analyzed by XRPD and the results are tabulated below (Table 12) and are shown in FIG. 5.  
                               TABLE 12                                       Intensity               d value (Å)   (%)   Intensity                                                        18.3   99   vs           12.5   22   s           9.9   22   s           9.1   67   vs           8.0   18   m           7.5   17   m           6.8   37   s           6.7   59   s           6.1   39   s           6.0   21   s           5.6   66   vs           5.5   98   vs           4.94   48   s           4.56   59   s           4.39   35   S           4.27   33   s           4.13   81   vs           4.02   87   vs           3.86   88   vs           3.69   69   vs           3.63   100   vs           3.57   49   s           3.48   53   s           3.23   35   s           3.19   43   s           3.16   38   s                      
 
      DSC showed an endotherm with an extrapolated melting onset temperature of ca 183° C. and TGA showed a 0.3% weight loss between 25-110° C.  
      Abbreviations  
     
         
          Ac=acetyl  
          APCI=atmospheric pressure chemical ionisation (in relation to MS)  
          API=atmospheric pressure ionisation (in relation to MS)  
          aq.=aqueous  
          Aze(&amp; (S)-Aze)=(S)-azetidine-2-carboxylate (unless otherwise specified)  
          Boc=tert-butyloxycarbonyl  
          br=broad (in relation to NMR)  
          CI=chemical ionisation (in relation to MS)  
          d=day(s)  
          d=doublet (in relation to NMR)  
          DCC=dicyclohexyl carbodiimide  
          dd=doublet of doublets (in relation to NMR)  
          DIBAL-H=di-isobutylaluminium hydride  
          DIPEA=diisopropylethylamine  
          DMAP=4-(N,N-dimethyl amino) pyridine  
          DMF=N,N-dimethylformamide  
          DMSO=dimethylsulfoxide  
          DSC=differential scanning colorimetry  
          DVT=deep vein thrombosis  
          EDC=1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride  
          eq.=equivalents  
          ES=electrospray  
          ESI=electrospray interface  
          Et=ethyl  
          ether=diethyl ether  
          EtOAc=ethyl acetate  
          EtOH=ethanol  
          Et 2 O=diethyl ether  
          HATU=O-(azabenzotriazol-1-yl)-N,N′,N′-tetramethyluronium hexafluorophosphate  
          HBTU=[N,N,N′,N′-tetramethyl-O-(benzotriazol-1-yl)uronium hexafluorophosphate] 
          HCl=hydrochloric acid, hydrogen chloride gas or hydrochloride salt (depending on context)  
          Hex=hexanes  
          HOAc=acetic acid  
          HPLC=high performance liquid chromatography  
          LC=liquid chromatography  
          m=multiplet (in relation to NMR)  
          Me=methyl  
          MeOH=methanol  
          min.=minute(s)  
          MS=mass spectroscopy  
          MTBE=methyl tert-butyl ether  
          NMR=nuclear magnetic resonance  
          OAc=acetate  
          Pab=para-amidinobenzylamino  
          H-Pab=para-amidinobenzylamine  
          Pd/C=palladium on carbon  
          Ph=phenyl  
          PyBOP=(benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate  
          q=quartet (in relation to NMR)  
          QF=tetrabutylammonium fluoride  
          rt/RT=room temperature  
          s=singlet (in relation to NMR)  
          solutol=PEG 660 12-hydroxy stearate (a non-ionic surfactant)  
          t=triplet (in relation to NMR)  
          TBTU=[N,N,N′,N′-tetramethyl-O-(benzotriazol-1-yl)uronium tetrafluoroborate] 
          TEA=triethylamine  
          Teoc=2-(trimethylsilyl)ethoxycarbonyl  
          TEMPO=2,2,6,6-tetramethyl-1-piperidinyloxy free radical  
          TFA=trifluoroacetic acid  
          TGA=thermogravimetric analysis  
          THF=tetrahydrofuran  
          TLC=thin layer chromatography  
          UV=ultraviolet  
       
    
      Prefixes n-, s-, i-, t- and tert- have their usual meanings: normal, secondary, iso, and tertiary.  
      The invention is illustrated by way of the following Examples. 
    
    
     EXAMPLE 1 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 30 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 50/5/45 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 50/5/45 (w/w) % followed by gently stirring. This composition was given to dogs orally by gavage once daily for 5 days. The dose 150 μmol/kg gave maximum plasma concentrations in the range 118-254 μM (118-254 μmol/L) after the first dose and 186-286 μM (186-286 μmol/L) after the fifth dose.  
     EXAMPLE 2 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 40 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 50/5/45 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 50/5/45 (w/w) % followed by gently stirring. This composition was given to rats orally by gavage once daily for 5 days. The dose 400 μmol/kg gave maximum plasma concentrations in the range 3.17-6.91 μM (3.17-6.91 μmol/L) after the first dose and 3.01-10.5 μM (3.01-10.5 μmol/L) after the fifth dose.  
     EXAMPLE 3 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 80 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 50/5/45 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 50/5/45 (w/w) % followed by gently stirring. This composition was given to rats orally by gavage once daily for 5 days. The dose 800 μmol/kg gave maximum plasma concentrations in the range 7.00-23.9 μM (7.00-23.9 μmol/L) after the first dose and 10.3-32.8 μM (10.3-32.8 μmol/L) after the fifth dose.  
     EXAMPLE 4 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 250 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 50/5/45 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 50/5/45 (w/w) % followed by gently stirring. The solubility of Compound A is at least 1000 times higher in this vehicle compared to water alone.  
     EXAMPLE 5 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 21 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 20/10/70 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 20/10/70 (w/w) % followed by gently stirring. The solubility of Compound A is at least 100 times higher in this vehicle compared to water alone.  
     EXAMPLE 6 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 51 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 20/10/70 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
               
                   
                   The water contained 50 μmol/mL Tartaric Acid    
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in acidified PEG 400/ethanol/water 20/10/70 (w/w) % that was followed by gently stirring. The pH of this solution was 3.6. The solubility of Compound A is at least 250 times higher in this vehicle compared to water alone.  
     EXAMPLE 7 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 44 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 30/5/65 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 30/5/65 (w/w) % followed by gently stirring. The solubility of Compound A is at least 200 times higher in this vehicle compared to water alone.  
     EXAMPLE 8 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 88 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 30/5/65 (w/w) % 
                 to 1 
                 mL 
               
            
           
           
               
               
               
            
               
                   
                 The water contained 50 μmol/mL Tartaric Acid 
                 q.s. 
               
               
                   
                 HCl to pH 3.6 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in acidified PEG 400/ethanol/water 30/5/65 (w/w) % followed by gently stirring. The pH of this solution was set to 3.6 by addition of HCl. The solubility of Compound A is at least 400 times higher in this vehicle compared to water alone.  
     EXAMPLE 9 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 120 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 40/5/55 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 40/5/55 (w/w) % followed by gently stirring. The solubility of Compound A is at least 600 times higher in this vehicle compared to water alone.  
     EXAMPLE 10 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 198 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 40/5/55 (w/w) % 
                 to 1 
                 mL 
               
            
           
           
               
               
               
            
               
                   
                 The water contained 50 μmol/mL Tartaric Acid 
                 q.s. 
               
               
                   
                 HCl to pH 3.8 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in acidified PEG 400/ethanol/water 40/5/55 (w/w) % followed by gently stirring. The pH of this solution was set to 3.8 by addition of HCl. The solubility of Compound A is at least 1000 times higher in this vehicle compared to water alone. Formulations of Compound A in this vehicle are stable for at least 3 months at &lt;−15° C.  
     EXAMPLE 11 
     
       
         
           
               
               
               
             
               
                   
               
               
                   
               
             
            
               
                 Compound A 
                 136 
                 μmol 
               
               
                 Hydroxypropyl-β-cyclodextrin/water 40/60 (w/w) % 
                 to 1 
                 mL 
               
            
           
           
               
               
            
               
                 HCl to pH 3.7 
                 q.s. 
               
               
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in Hydroxypropyl-β-cyclodextrin/water 40/60 (w/w) % followed by gently stirring. The pH of this solution was set to 4.7 by addition of HCl. The solubility of Compound A is at least 700 times higher in this vehicle compared to water alone.  
     EXAMPLE 12 
     
       
         
           
               
               
               
             
               
                   
               
               
                   
               
             
            
               
                 Compound A 
                 76 
                 μmol 
               
               
                 Hydroxypropyl-β-cyclodextrin/water 28/72 (w/w) % 
                 to 1 
                 mL 
               
               
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in Hydroxypropyl-β-cyclodextrin/water 28/72 (w/w) % followed by gently stirring. The solubility of Compound A is at least 400 times higher in this vehicle compared to water alone.  
     EXAMPLE 13 
     
       
         
           
               
               
               
             
               
                   
               
               
                   
               
             
            
               
                 Compound A 
                 40 
                 μmol 
               
               
                 PEG 400/ethanol/solutol ™/water 50/5/5/40 (w/w) % 
                 to 1 
                 mL 
               
               
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/solutol™/water 50/5/5/40 (w/w) % followed by gently stirring. The solubility of Compound A is at least 80 times higher in this vehicle compared to water alone.  
     EXAMPLE 14 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 40 
                 μmol 
               
               
                   
                 PEG 400/water 40/60 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400 followed by gently stirring for at least 1 hour, thereafter water was added to the final volume. The solubility of Compound A is at least 200 times higher in this vehicle compared to water alone.  
     EXAMPLE 15 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 52 
                 μmol 
               
               
                   
                 PEG 400/water 35/65 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                 The water contained 50 μmol/mL Tartaric Acid 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400 followed by gently stirring for at least 1 hour, thereafter water was added to the final volume. The solubility of Compound A is at least 250 times higher in this vehicle compared to water alone.  
     EXAMPLE 16 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 58 
                 μmol 
               
               
                   
                 PEG 400/water 50/50 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400 followed by gently stirring for at least 1 hour, thereafter water was added to the final volume. The solubility of Compound A is at least 300 times higher in this vehicle compared to water alone.  
     EXAMPLE 17 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 88 
                 μmol 
               
               
                   
                 PEG 400/water 67/33 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400 followed by gently stirring for at least 1 hour, thereafter water was added to the final volume. The solubility of Compound A is at least 400 times higher in this vehicle compared to water alone.  
     EXAMPLE 18 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 92 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 45/1/54 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 45/1/54 (w/w) % followed by gently stirring. The solubility of Compound A is at least 450 times higher in this vehicle compared to water alone.  
     EXAMPLE 19 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 159 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 45/1/54 (w/w) % 
                 to 1 
                 mL 
               
            
           
           
               
               
               
            
               
                   
                 The water contained 50 μmol/mL Tartaric Acid 
                 q.s. 
               
               
                   
                 HCl to pH 4.2 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in acidified PEG 400/ethanol/water 45/1/54 (w/w) % followed by gently stirring. The pH of this solution was set to 4.2 with HCl. The solubility of Compound A is at least 800 times higher in this vehicle compared to water alone.  
     EXAMPLE 20 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 101 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 45/2/53 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 45/2/53 (w/w) % followed by gently stirring. The solubility of Compound A is at least 500 times higher in this vehicle compared to water alone.  
     EXAMPLE 21 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 167 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 45/2/53 (w/w) % 
                 to 1 
                 mL 
               
            
           
           
               
               
               
            
               
                   
                 The water contained 50 μmol/mL Tartaric Acid 
                 q.s. 
               
               
                   
                 HCl to pH 4.3 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in acidified PEG 400/ethanol/water 45/2/53 (w/w) % followed by gently stirring. The pH of this solution was set to 4.3 by addition of HCl. The solubility of Compound A is at least 800 times higher in this vehicle compared to water alone.  
     EXAMPLE 22 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 46 
                 μmol 
               
               
                   
                 DMA/water 50/50 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in the vehicle followed by gently stirring for at least 1 hour. The solubility of Compound A is at least 230 times higher in this vehicle compared to water alone.  
     EXAMPLE 23 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 29 
                 μmol 
               
               
                   
                 DMA/water 25/75 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in the vehicle followed by gently stirring for at least 1 hour. The solubility of Compound A is at least 150 times higher in this vehicle compared to water alone.  
     EXAMPLE 24 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 5 
                 μmol 
               
               
                   
                 HCl 
                 10 
                 μmol 
               
               
                   
                 Water 
                 to 1 
                 mL 
               
            
           
           
               
               
               
            
               
                   
                 HCl/NaOH to pH 3.6 
                 q.s. 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in a lower volume of the double equimolar amount of HCl followed by gently stirring and dilution to 1 mL. The pH of the final solution was adjusted to 3.6. The solubility of Compound A is at least 20 times higher in this vehicle compared to water alone.  
     EXAMPLE 25 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 10 
                 μmol 
               
               
                   
                 Water 
                 to 1 
                 mL 
               
            
           
           
               
               
               
            
               
                   
                 HCl to pH 1.0 
                 q.s. 
               
               
                   
                 NaOH to pH 3.0 
                 q.s. 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A water and HCl was added to give pH 1 thereafter the solution was gently stirred. The pH of the final solution was adjusted to 3.0 with NaOH. The solubility of Compound A is at least 40 times higher in this vehicle compared to water alone. This formulation was given p.o to rats in a kinetic comparative study.  
     EXAMPLE 26 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 100 
                 μmol 
               
               
                   
                 Miglyol 
                 0.25 
                 g/g Compound A 
               
               
                   
                 DMA 
                 to 
                 1 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in 1 mL DMA/miglyol followed by gently stirring. The solubility of Compound A is at least 4000 times higher in this vehicle compared to water alone.  
     EXAMPLE 27 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 100 
                 μmol 
               
               
                   
                 Miglyol 
                 0.25 
                 g/g Compound A 
               
               
                   
                 Ethanol 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in 1 mL Ethanol/Miglyol followed by gently stirring. The solubility of Compound A is at least 4000 times higher in this vehicle compared to water alone.  
     EXAMPLE 28 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 130 
                 μmol 
               
               
                   
                 Ethanol 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in 1 mL ethanol followed by gently stirring. The substance is stable in this formulation more than 1 week.  
     EXAMPLE 29  
      In order to prepare nanoparticles a stock solution of Compound A of about 100 mM in ethanol was used. Included was also 25% (w/w) Miglyol, calculated on the amount of the substance. The solutions were diluted 1/10 with the stabilizer solution, consisting of 0.2% (w/w) PVP and 0.25 mM SDS in water. The mixing, which is considered as a critical parameter during the nanoparticle preparation, was rapid and instant. The drug solution was rapidly injected into the stabilizer solution during ultrasonication. After the 1/10 dilution in the aqeous solution, nanoparticles of about 150 nm were achieved. After 6 hours at room temperature, the particle sizes were unchanged.  
     EXAMPLE 30 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 4 
                 μmol 
               
               
                   
                 saline/ethanol/solutol 90/5/5 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in saline/ethanol/solutol 90/5/5 (w/w) % followed by gently stirring. The solution was given orally to rats and the plasma concentration of Compound D was 0.56 μmol/L after 1 hour. The solution was given subcutaneously to rats and the plasma concentrations of Compound D and A were 0.24 μmol/L and 0.6 μmol/L, respectively, after 1 hour.  
     EXAMPLE 31 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound B 
                 4 
                 μmol 
               
               
                   
                 saline/ethanol/solutol 90/5/5 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound B in saline/ethanol/solutol 90/5/5 (w/w) % followed by gently stirring. The solution was given orally to rats and the plasma concentrations of Compound B and Compound E were respectively 0.07 μmol/L and 0.65 μmol/L, after 1 hour. The solution was given subcutaneously to rats and the plasma concentrations of Compound B and E were 0.4 μmol/L and 0.3 μmol/L, respectively, after 1 hour.  
     EXAMPLE 32 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound C 
                 4 
                 μmol 
               
               
                   
                 saline/ethanol/solutol 90/5/5 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound C in saline/ethanol/solutol 90/5/5 (w/w) % followed by gently stirring. The solution was given orally to rats and the plasma concentrations of Compounds C and F were respectively 0.2 μmol/L and 0.5 μmol/L after 1 hour. The solution was given subcutaneously to rats and the plasma concentrations of Compounds C and F were 0.35 μmol/L and 0.5 μmol/L, respectively, after 1 hour.  
     EXAMPLE 33 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound D (trifluoroacetate salt) 
                 5 
                 μmol 
               
               
                   
                 Saline 9 mg/ml 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving the salt of Compound D in 1 mL saline followed by gently stirring.  
     EXAMPLE 34 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound D (trifluoroacetate salt) 
                 75 
                 μmol 
               
               
                   
                 EtOH 
                 0.05 
                 mL 
               
               
                   
                 Saline(9 mg/ml) 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving the salt of Compound D in 1 mL saline/ethanol solution followed by gently stirring.  
     EXAMPLE 35 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound D (trifluoroacetate salt) 
                 4 
                 μmol 
               
               
                   
                 EtOH 
                 0.02 
                 mL 
               
               
                   
                 saline 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving the salt of Compound D in 1 mL saline/etanol solution followed by gently stirring. The solution was given subcutaneously to rats and the plasma concentration of Compound D was 0.55 μmol/L after 1 hour.  
     EXAMPLE 36 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound E (acetate salt) 
                 4 
                 μmol 
               
               
                   
                 EtOH 
                 0.02 
                 mL 
               
               
                   
                 saline 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving the salt of Compound E in 1 mL saline/ethanol solution followed by gently stirring. The solution was given subcutaneously to rats and the plasma concentration of Compound E was 0.75 μmol/L after 1 hour.  
     EXAMPLE 37 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound F (trifluoroacetate salt) 
                 4 
                 μmol 
               
               
                   
                 EtOH 
                 0.02 
                 mL 
               
               
                   
                 saline 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving the salt of Compound F in 1 mL saline/ethanol solution followed by gently stirring. The solution was given subcutaneously to rats and the plasma concentration Compound F was 0.92 μmol/L after 1 hour.  
     EXAMPLE 38 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound E (acetate salt) 
                 22 
                 mg 
               
               
                   
                 Saline 9 mg/ml 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving the salt of Compound E in 1 mL saline followed by gently stirring.  
     EXAMPLE 39 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound F (trifluoroacetate salt) 
                 22 
                 mg 
               
               
                   
                 Saline 9 mg/ml 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving the salt of Compound F in 1 mL saline followed by gently stirring.  
     EXAMPLE 40 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A (as esylate salt) 
                 14 
                 mg 
               
               
                   
                 water 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A solution was prepared by dissolving excess of Compound A as esylate salt in 3 mL water followed by gently stirring over night. A final concentration of the solution after filtration was monitored to 14 mg/ml at a pH of 2.7.  
     EXAMPLE 41 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A (as esylate salt) 
                 33 
                 mg 
               
               
                   
                 Sodium phosphate buffer pH = 3.1 I = 0.1 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A solution was prepared by dissolving 112 mg of Compound A as esylate salt in 3 mL sodium phosphate buffer followed by gently stirring over night. A final concentration of the solution after filtration was monitored to 33 mg/ml at a pH of 2.7.  
     EXAMPLE 42 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A (as esylate salt) 
                 1.6 
                 mg 
               
               
                   
                 Sodium phosphate buffer pH = 6.9 I = 0.1 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A solution was prepared by dissolving 20 mg of Compound A as esylate salt in 3 mL sodium phosphate buffer followed by gently stirring over night. A final concentration of the solution after filtration was monitored to 1.6 mg/ml at a pH of 6.5.  
     EXAMPLE 43  
      The following freeze dried formulations can be made in accordance with techniques described in one or more of Examples 1-29 above:  
                                                          a.                   Compound A   10   μmol           Mannitol   10   mg           Water   to 1   mL                             HCl to pH 1.0   q.s.           NaOH to pH 3.0   q.s.                                 b.                   Compound D   10   μmol           Mannitol   10   mg           Water   to 1   mL                             HCl to pH 1.0   q.s.           NaOH to pH 3.0   q.s.                                 c.                   Compound E   10   μmol           Mannitol   10   mg           Water   to 1   mL                             HCl to pH 1.0   q.s.           NaOH to pH 3.0   q.s.                                 d.                   Compound F   10   μmol           Mannitol   10   mg           Water   to 1   mL                             HCl to pH 1.0   q.s.           NaOH to pH 3.0   q.s.                                 e.                   Compound B   10   μmol           Mannitol   10   mg           Water   to 1   mL                             HCl to pH 1.0   q.s.           NaOH to pH 3.0   q.s.                                 f.                   Compound C   10   μmol           Mannitol   10   mg           Water   to 1   mL                             HCl to pH 1.0   q.s.           NaOH to pH 3.0   q.s.                                 g.                   Compound A (as esylate salt)   14   mg           Mannitol   10   mg           Water   to 1   mL                             HCl to pH 1.0   q.s.           NaOH to pH 3.0   q.s.                                 h.                   Compound A (as besylate salt)   14   mg           Mannitol   10   mg           Water   to 1   mL                             HCl to pH 1.0   q.s.           NaOH to pH 3.0   q.s.                      
 
      The solutions are optionally sterile filtered, for example through a 0.22 μm membrane filter. Solutions (sterile or otherwise) are filled into appropriate vessels (e.g. vials) and the formulations are freeze-died using standard equipment. Vials may be sealed in freeze-dryer equipment under a nitrogen atmosphere.  
     EXAMPLE 44 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                   
               
               
                   
                 Weight 
                 Amount 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Compound A 
                 48 mg 
                 17% 
               
               
                   
                 Polyvinyl pyrrolidone K90 
                  8 mg 
                 3% 
               
               
                   
                 Mannitol 
                 21 mg 
                 7% 
               
               
                   
                 Microcrystalline cellulose 
                 187 mg  
                 65% 
               
               
                   
                 Sodium starch glycolate 
                 21 mg 
                 7% 
               
               
                   
                 Sodium stearyl fumarate 
                  3 mg 
                 1% 
               
               
                   
                   
               
            
           
         
       
     
      The excipients and drug were mixed and granulated with polyvinyl pyrrolidone K90 dissolved in water. The granules were then dried in a drying oven. The granulate was lubricated with sodiumstearylfumarate and compressed into tablets using an excenterpress.  
      Three individual tablets were tested for drug release in 900 ml media using a USP dissolution apparatus 2 (paddle+basket 1 ) at 50 rpm and 37° C. The dissolution media used were 0.1 M hydrochloric acid (pH 1) and 0.1 M sodium phosphate buffer (pH 6.8). In-line quantitation was performed using the C Technologies fibre optic system with 220 nm as the analytical wavelength when 0.1 M HCl was used as the dissolution media and with 260 nm as the analytical wavelength when phosphate buffer pH 6.8 was used as the dissolution media. 350 nm was used as the reference wavelength with both media. For the first two hours of the analysis the release value was measured every 15 minutes, and then every hour for the remainder of the analysis. The results are presented in the table below.  [ 1  A custom made quadrangular basket of mesh wire, soldered in one of its upper, narrow sides to the end of a steel rod. The rod is brought through the cover of the dissolution vessel and fixed by means of two Teflon nuts, 3.2 cm from the centre of the vessel. The lower edge of the bottom of the basket is adjusted to be 1 cm above the paddle. The basket is directed along the flow stream with the tablet under test standing on its edge].   
                                           % released in   % released in       Time (min)   buffer pH 1.1   buffer pH 6.8                                            0   0   0       15   100   44       30   100   49       45   100   51       60   100   53       120   100   57       180   100   61       240   100   63       360   100   67       480   100   70       600   100   75       720   100   77       840   100   79       960   100   82       1080   100   83       1200   100   86                  
 
     EXAMPLE 45 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                   
               
               
                   
                 Weight 
                 Amount 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Esylate salt of Compound A 
                 58 mg 
                 20% 
               
               
                   
                 Polyvinyl pyrrolidone K90 
                  8 mg 
                 3% 
               
               
                   
                 Mannitol 
                 21 mg 
                 7% 
               
               
                   
                 Microcrystalline cellulose 
                 177 mg  
                 62% 
               
               
                   
                 Sodium starch glycolate 
                 21 mg 
                 7% 
               
               
                   
                 Sodium stearyl fumarate 
                  3 mg 
                 1% 
               
               
                   
                   
               
            
           
         
       
     
      The excipients and drug were mixed and granulated with polyvinyl pyrrolidone K90 dissolved in water. The granules were then dried in a drying oven. The granulate was lubricated with sodium stearyl fumarate and compressed into tablets using an excenterpress.  
     EXAMPLE 46 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                   
               
               
                   
                 Weight 
                 Amount 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Compound B 
                 48 mg 
                 17% 
               
               
                   
                 Polyvinyl pyrrolidone K90 
                  8 mg 
                 3% 
               
               
                   
                 Mannitol 
                 21 mg 
                 7% 
               
               
                   
                 Microcrystalline cellulose 
                 187 mg  
                 65% 
               
               
                   
                 Sodium starch glycolate 
                 21 mg 
                 7% 
               
               
                   
                 Sodium stearyl fumarate 
                  3 mg 
                 1% 
               
               
                   
                   
               
            
           
         
       
     
      The excipients and drug were mixed and granulated with polyvinyl pyrrolidone K90 dissolved in water. The granules were then dried in a drying oven. The granulate was lubricated with sodium stearyl fumarate and compressed into tablets using an excenterpress  
     EXAMPLE 47 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                   
               
               
                   
                 Weight 
                 Amount 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Compound C 
                 48 mg 
                 17% 
               
               
                   
                 Polyvinyl pyrrolidone K90 
                  8 mg 
                 3% 
               
               
                   
                 Mannitol 
                 21 mg 
                 7% 
               
               
                   
                 Microcrystalline cellulose 
                 187 mg  
                 65% 
               
               
                   
                 Sodium starch glycolate 
                 21 mg 
                 7% 
               
               
                   
                 Sodium stearyl fumarate 
                  3 mg 
                 1% 
               
               
                   
                   
               
            
           
         
       
     
      The excipients and drug were mixed and granulated with polyvinyl pyrrolicdone K90 dissolved in water. The granules were then dried in a drying oven. The granulate was lubricated with sodium stearyl fumarate and compressed into tablets using an excenterpress  
     EXAMPLE 48 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 16 
                 μmol 
               
               
                   
                 PEG 414 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in acidified PEG414 followed by gently stirring.  
     EXAMPLE 49 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 16 
                 μmol 
               
               
                   
                 PEG 300 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in acidified PEG300 followed by gently stirring.  
     EXAMPLE 50 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 16 
                 μmol 
               
               
                   
                 PEG 200 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in acidified PEG200 followed by gently stirring.  
     EXAMPLE 51 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound G 
                 4 
                 μmol 
               
               
                   
                 saline/ethanol/solutol 90/5/5 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound G in saline/ethanol/solutol 90/5/5 (w/w) % followed by gently stirring.  
     EXAMPLE 52 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound J 
                 4 
                 μmol 
               
               
                   
                 saline/ethanol/solutol 90/5/5 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound J in saline/ethanol/solutol 90/5/5 (w/w) % followed by gently stirring.  
     EXAMPLE 53 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound H 
                 4 
                 μmol 
               
               
                   
                 saline/ethanol/solutol 90/5/5 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound H in saline/ethanol/solutol 90/5/5 (w/w) % followed by gently stirring.  
     EXAMPLE 54 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                   
               
               
                   
                 Weight 
                 Amount 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Compound A esylate salt 
                 500 mg 
                 66% 
               
               
                   
                 Polyvinyl pyrrolidone K30 
                 100 mg 
                 13% 
               
               
                   
                 Microcrystalline cellulose 
                 100 mg 
                 13% 
               
               
                   
                 Crosslinked sodium CMC 
                  50 mg 
                 7% 
               
               
                   
                 Magnesium stearate 
                  5 mg 
                 1% 
               
               
                   
                   
               
            
           
         
       
     
      Formulation can be prepared in accordance with Example 47 above.  
     EXAMPLE 55 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                   
               
               
                   
                 Weight 
                 Amount 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Compound A n-propane 
                 100 mg 
                 23% 
               
               
                   
                 sulphonic acid salt 
               
               
                   
                 Polyvinyl pyrrolidone K30 
                  60 mg 
                 14% 
               
               
                   
                 Lactose monohydrate 
                 100 mg 
                 23% 
               
               
                   
                 Microcrystalline cellulose 
                 150 mg 
                 34% 
               
               
                   
                 Polyvinyl pyrrolidone 
                  20 mg 
                 5% 
               
               
                   
                 crosslinked 
               
               
                   
                 Sodium stearyl fumarate 
                  10 mg 
                 2% 
               
               
                   
                   
               
            
           
         
       
     
      Formulation can be prepared in accordance with Example 47 above.  
     EXAMPLE 56 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                   
               
               
                   
                 Weight 
                 Amount 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Compound A besylate salt 
                 20 mg 
                 8% 
               
               
                   
                 Hydroxypropyl cellulose 
                 15 mg 
                 6% 
               
               
                   
                 Microcrystalline cellulose 
                 200 mg  
                 79% 
               
               
                   
                 Crosslinked sodium CMC 
                 15 mg 
                 6% 
               
               
                   
                 Sodium stearyl fumarate 
                  3 mg 
                 1% 
               
               
                   
                   
               
            
           
         
       
     
      Formulation can be prepared in accordance with Example 47 above.  
     EXAMPLE 57 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 24 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 25/10/65 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 25/10/65 (w/w) % followed by gently stirring. The solubility of Compound A is at least 100 times higher in this vehicle compared to water alone. The formulation is stable in a freezer for at least 2 months.  
     EXAMPLE 58 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 800 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 50/10/40 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 50/10/40 (w/w) % followed by gently stirring. The solubility of Compound A is at least 2000 times higher in this vehicle compared to water alone.  
     EXAMPLE 59 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 500 
                 μmol 
               
               
                   
                 Citric acid 
                 200 
                 μmol 
               
            
           
           
               
               
               
            
               
                   
                 HCl to pH 3.6 
                 q.s. 
               
            
           
           
               
               
               
               
            
               
                   
                 PEG 400/ethanol/9 mg/ml NaCl 40/10/50 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 40/10/50 (w/w) % followed by gently stirring. The solubility of Compound A is at least 1500 times higher in this vehicle compared to water alone.  
     EXAMPLE 60 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 24 
                 μmol 
               
               
                   
                 citric acid 
                 5 
                 μmol 
               
            
           
           
               
               
               
            
               
                   
                 HCl to pH 3.2 
                 q.s. 
               
            
           
           
               
               
               
               
            
               
                   
                 ethanol/water 12/88 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in ethanol followed by gently stirring, thereafter citric acid and water was added to final volume and the pH was set to 3.2. The solubility of Compound A is at least 100 times higher in this vehicle compared to water alone. The formulation is stable in a freezer for at least 1 month.  
     EXAMPLE 61 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 2 
                 μmol 
               
               
                   
                 citric acid 
                 5 
                 μmol 
               
            
           
           
               
               
               
            
               
                   
                 HCl to pH 3.6 
                 q.s. 
               
            
           
           
               
               
               
               
            
               
                   
                 9 mg/ml NaCl 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A and citric acid in physicological saline followed by gently stirring. The pH was set to 3.6. The formulation is stable in a freezer for at least 3 months.  
     EXAMPLE 62 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A (as besylate salt) 
                 140 
                 μmol 
               
               
                   
                 citric acid 
                 5 
                 μmol 
               
            
           
           
               
               
               
            
               
                   
                 HCl to pH 3.6 
                 q.s. 
               
            
           
           
               
               
               
               
            
               
                   
                 PEG 400/ethanol/water 40/5/55 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 40/5/55 (w/w) % containing citric acid followed by gently stirring and setting the pH to 3.6. The formulation is stable in a freezer for at least 1 month.  
     EXAMPLE 63 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A (as besylate salt) 
                 65 
                 μmol 
               
               
                   
                 citric acid 
                 5 
                 μmol 
               
            
           
           
               
               
               
            
               
                   
                 HCl to pH 3.3 
                 q.s. 
               
            
           
           
               
               
               
               
            
               
                   
                 PEG 400/ethanol/water 20/5/75 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A in PEG 400/ethanol/water 20/5/75 (w/w) % containing citric acid followed by gently stirring and the pH was set to 3.2.  
     EXAMPLE 64 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound D (as acetate salt) 
                 25 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 40/5/55 (w/w) % 
                 to 1 
                 mL 
               
            
           
           
               
               
               
            
               
                   
                 Tartaric Acid: Component A (acetate salt of D) 
                 q.s. 
               
               
                   
                 equimolar amount plus 5 mM excess 
               
               
                   
                 HCl to pH 3.6 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound D in acidified PEG 400/ethanol/water 40/5/55 (w/w) % followed by gently stirring. The pH of this solution was set to 3.6 by addition of HCl. Formulations of D in this vehicle are stable for at least 2 months at &lt;−15° C.  
     EXAMPLE 65 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A 
                 50 
                 mg 
               
               
                   
                 HPMC (15000 Cps) 
                 5 
                 mg 
               
               
                   
                 Solutol HS15 
                 20 
                 mg 
               
               
                   
                 Water 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      The HPMC was suspended in hot water and melted Solutol was added during vigourous stirring. This solution was chilled and Compound A was added under vigourous stirring to form a well dispersed suspension.  
     EXAMPLE 66 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound A (as besylate salt) 
                 50 
                 mg 
               
               
                   
                 HPMC (15000 Cps) 
                 5 
                 mg 
               
               
                   
                 Solutol HS15 
                 20 
                 mg 
               
               
                   
                 Water 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      The HPMC was suspended in hot water and melted Solutol was added during vigourous stirring. This solution was shilled and Compound A (besylate) was added under vigourous stirring to form a well dispersed suspension.  
     EXAMPLE 67 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound D (as acetate salt) 
                 2 
                 μmol 
               
               
                   
                 citric acid 
                 5 
                 μmol 
               
            
           
           
               
               
               
            
               
                   
                 HCl to pH 3.6 
                 q.s. 
               
            
           
           
               
               
               
               
            
               
                   
                 9 mg/ml NaCl 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound A and citric acid in physicological saline and stirring gently. The pH was set to 3.6. The formulation is stable in a freezer for at least 3 months.  
     EXAMPLE 68  
      To prepare nanoparticles a stock solution of Compound B of about 100 mM in ethanol was used. Included was also 25% (w/w) Miglyol, calculated on the amount of the substance. The solutions were diluted 1/10 with a stabilizer solution consisting of 0.2% (w/w) PVP and 0.25 mM SDS in water. The critical mixing stage was rapid and instant:—The drug solution was rapidly injected into the stabilizer solution during ultrasonication. After 1/10 dilution in the aqeous solution, nanoparticles of about 110 nm were obtained. After 6 hours at room temperature, the particle sizes were unchanged.  
      Optionally DMA may be used instead of ethanol, Miglyol may be excluded and the dilution may be larger (1/20). Particles in the size range 100 to 300 nm may be obtained by different combinations.  
     EXAMPLE 69 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound B 
                 200 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 50/5/45 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound B in PEG 400/ethanol/water 50/5/45 (w/w) % followed by gently stirring. Formulations of B (at 0.5 mg/mL) in this vehicle are stable for at least 1 month at &lt;−15° C.  
     EXAMPLE 70 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound B 
                 230 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 60/5/35 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound B in PEG 400/ethanol/60/5/35 (w/w) % followed by gently stirring.  
     EXAMPLE 71 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound B 
                 50 
                 mg 
               
               
                   
                 HPMC (15000 Cps) 
                 5 
                 mg 
               
               
                   
                 Solutol HS15 
                 20 
                 mg 
               
               
                   
                 Water 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      The HPMC was suspended in hot water and melted solutol was added during vigourous stirring. This solution was chilled and Compound B was added under vigourous stirring to form a well dispersed suspension.  
     EXAMPLE 72 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound E (as acetate salt) 
                 39 
                 μmol 
               
               
                   
                 9 mg/ml NaCl 
                 to 1 
                 mL  
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound E in 9 mg/ml NaCl by gently stirring. The pH obtained in this formulation is 8-9.  
     EXAMPLE 73 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound C 
                 400 
                 μmol 
               
               
                   
                 PEG 400/ethanol/water 50/5/45 (w/w) % 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound C in PEG 400/ethanol/water 50/5/45 (w/w) % followed by gently stirring. Formulations of C (at 0.5 mg/mL) in this vehicle are stable for at least 1 month at room temperature and below.  
     EXAMPLE 74 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound C 
                 16 
                 μmol 
               
               
                   
                 Hydroxypropyl-β-cyclodextrin/water 20/80 (w/w) % 
                 to 1 
                 mL  
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound C in Hydroxypropyl-β-cyclodextrin/water 20/80 (w/w) % followed by gently stirring. Formulations of C in this vehicle are stable for at least 2 weeks at &lt;8° C.  
     EXAMPLE 75 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
               
             
            
               
                   
                 Compound F (as trifluoroacetate salt) 
                 38 
                 μmol 
               
               
                   
                 9 mg/ml NaCl 
                 to 1 
                 mL 
               
               
                   
                   
               
            
           
         
       
     
      A formulation was prepared by dissolving Compound F in 9 mg/ml NaCl by gently stirring. The pH obtained in this formulation is 3-4. Formulations of F in this vehicle are stable for at least 2 weeks at at room temperature and below.  
     EXAMPLE 76  
      A tablet was prepared according to the general method of Example 44.  
                                                           Weight   Amount                                                        Besylate salt of Compound A   66 mg   17%           Polyvinyl pyrrolidone K90    9 mg   2%           Mannitol   29 mg   7%           Microcrystalline cellulose   256 mg    65%           Sodium starch glycolate   29 mg   7%           Sodium stearyl fumarate    4 mg   1%                      
 
     Release Data  
      Measured according to the general method of Example 44 but using 500 ml of media and 75 rpm.  
                                                       % released in           Time (min)   buffer pH 6.8                                                    0   0           5   90           10   94           15   96           20   96           30   98           45   98           60   100                      
 
     EXAMPLE 77  
      A tablet is prepared according to the general method of Example 44.  
                                                           Weight   Amount                                                        Besylate salt of Compound A   200 mg    40%           Polyvinyl pyrrolidone K30   10 mg   2%           Lactose   200 mg    40%           Microcrystalline cellulose   70 mg   14%           Polyvinylpolypyrrolidone CL   15 mg   3%           Magnesium stearate    5 mg   1%                      
 
      Other formulations in which the quantity of the besylate salt of Compound A is in the range 50-300 mg may be prepared; the ratio of other components being similar to those in Example 77.  
     EXAMPLE 78  
      A tablet is prepared according to the general method of Example 44.  
                                                           Weight   Amount                                                        Hemi-Naphthalene 1,5-disulphonic   48 mg   17%           acid salt of Compound B           Polyvinyl pyrrolidone K90    8 mg   3%           Mannitol   21 mg   7%           Microcrystalline cellulose   187 mg    65%           Sodium starch glycolate   21 mg   7%           Sodium stearyl fumarate    3 mg   1%                      
 
      Other formulations in which 100 mg or 200 mg of the hemi-naphthalene 1,5-disulphonic acid salt of Compound B is used may also be prepared; the ratio of other components being similar to those in Example 78.  
      Particular aspects of the invention are provided as follows:—
      1. An immediate release pharmaceutical formulation comprising, as active ingredient, a compound of formula (I):  
                 
        wherein     R 1  represents C 1-2  alkyl substituted by one or more fluoro substituents;     R 2  represents hydrogen, hydroxy, methoxy or ethoxy; and     n represents 0, 1 or 2;     or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable diluent or carrier;     provided that the formulation does not solely contain: 
            a solution of one active ingredient and water;     a solution of one active ingredient and dimethylsulphoxide; or,     a solution of one active ingredient in a mixture of ethanol:PEG 660 12-hydroxy stearate:water 5:5:90.    
           
        2. An immediate release pharmaceutical formulation as described in aspect 1 wherein the active ingredient is:     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(OMe);     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF)(OMe);     Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)—(S)Aze-Pab(OMe);     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab;     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(OH);     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF);     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF)(OH);     Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)—(S)Aze-Pab; or,     Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)—(S)Aze-Pab(OH).     3. A solid immediate release pharmaceutical formulation as described in aspect 1 wherein the active ingredient is:     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(OMe);     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF)(OMe); or,     Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)—(S)Aze-Pab(OMe),     or a pharmaceutically acceptable salt thereof.     4. A solid immediate release pharmaceutical formulation as described in aspect 1 wherein the active ingredient is Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(OMe) or a C 1-6  alkanesulfonic acid or an optionally substituted arylsulfonic acid salt thereof.     5. An injectable immediate release pharmaceutical formulation as described in aspect 1 wherein the active ingredient is:     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab;     Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF); or     Ph(3-Cl)(5-OCH 2 CH 2 F)—(R)CH(OH)C(O)—(S)Aze-Pab.     6. The use of a formulation as described in aspect 1 as a medicament.     7. The use of a formulation as described in aspect 1 in the manufacture of a medicament for the treatment of a cardiovascular disorder.     8. A method of treating a cardiovascular disorder in a patient suffering from, or at risk of, said disorder, which comprises administering to the patient a therapeutically effective amount of a pharmaceutical formulation as described in aspect 1.     9. A process for making an immediate release formulation as described in aspect 1.     10. The compound Ph(3-Cl)(5-OCHF 2 )—(R)CH(OH)C(O)—(S)Aze-Pab(2,6-diF)(OH).    

      Also provided is a formulation obtainable by any of the Methods and/or Examples described herein.