Crystalline Eravacycline Bis-Hydrochloride

Abstract

The invention relates to crystalline eravacycline Ns-hydrochloride and to a process for its preparation. Furthermore, the invention relates to the use of crystalline eravacycline bis-hydrochloride for the preparation of pharmaceutical compositions. The invention further relates to pharmaceutical compositions comprising an effective amount of crystalline eravacycline bis-hydrochloride. The pharmaceutical compositions of the present invention can be used as medicaments, in particular for treatment and/or prevention of bacterial infections e.g. caused by Gram negative pathogens or Gram positive pathogens, in particular caused by multidrug resistant Gram negative pathogens. The pharmaceutical compositions of the present invention can thus be used as medicaments for e.g. the treatment of complicated intra-abdominal and urinary tract infection

1 - 13 . (canceled) 
     
     
         14 . A process for the preparation of crystalline eravacycline bis-hydrochloride of formula (III) 
       
         
           
           
               
               
           
         
       
       or a solvate or a hydrate thereof,
 wherein n is in the range from 1.8 to 2.2, and 
 wherein m is in the range from about 0.0 to 9.0, the process comprising: 
 (i) providing an aqueous solution of eravacycline bis-hydrochloride in water and at least one organic antisolvent selected from the group consisting of ketones and ethers, wherein the aqueous solution is characterized by a water activity of at least 0.1 and, 
 (ii) adding eravacycline bis-hydrochloride seed crystals to the solution provided in (i), wherein the seed crystals of (ii) are prepared according to a process comprising step A, or step B, or step C:
 Step A:
 a) Preparing a suspension of amorphous eravacycline bis-hydrochloride in 1-propanol and stirring, while cycling between a temperature of 25° C. and 0° C.; and 
 b) Isolating the solid of the suspension, thereby obtaining a sample containing seed crystals of eravacycline bis-hydrochloride; or 
 
 Step B:
 a) Dissolving amorphous eravacycline bis-hydrochloride in aqueous hydrochloric acid, thereby obtaining a solution; 
 b) Adding acetone to the solution until a milky solution is obtained; 
 c) Seeding the solution of step B-b) with the eravacycline bis-hydrochloride crystals of step A; 
 d) Storing the mixture of step B-c) at a temperature of 0 to 5° C., thereby obtaining a suspension; 
 e) Adding acetone to the suspension of step B-d); 
 f) Storing the mixture of step B-e) at a temperature of 0 to 5° C.; 
 g) Adding acetone to the mixture of step B-f), thereby obtaining a solid; and 
 h) Isolating the solid of step B-g), thereby obtaining the seed crystal of eravacycline bis-hydrochloride; or 
 
 Step C:
 a) Suspending amorphous eravacycline free base in water; 
 b) Adding concentrated hydrochloric acid, thereby obtaining a clear solution; 
 c) Adding acetone to the solution of step b), thereby obtaining a milky solution; 
 d) Seeding the milky solution with the seed crystal of step A-b) or of step B-h); 
 e) Adding acetone to the seeded solution of step C-d), thereby obtaining a suspension; 
 f) Stirring the suspension of step C-e), thereby obtaining a solid; and 
 g) Isolating the solid of step C-f), thereby obtaining the seed crystal of eravacycline bis hydrochloride. 
 
 
 
     
     
         15 . Process for the preparation of crystalline eravacycline bis-hydrochloride of formula III of  claim 14 , wherein in Step A-a) the cycling comprises stirring the suspension at 25° C., cooling the suspension to 0° C., stirring the suspension at 0° C., and heating the suspension to 25° C. 
     
     
         16 . Process for the preparation of crystalline eravacycline bis-hydrochloride of  claim 15 , wherein the stirring step, the cooling step, and the heating step are each carried out for about 1 hour. 
     
     
         17 . Process for the preparation of crystalline eravacycline bis-hydrochloride of  claim 16 , wherein the cycling step is repeated over a period of about 48 hours in total. 
     
     
         18 . Process for the preparation of crystalline eravacycline bis-hydrochloride of formula III of  claim 14 , wherein in Step A-b) the isolating step is carried out by centrifugation. 
     
     
         19 . Process for the preparation of crystalline eravacycline bis-hydrochloride of  claim 14 , wherein in Step B-a) the amorphous eravacycline bis-hydrochloride is obtained according to Step A. 
     
     
         20 . Process for the preparation of crystalline eravacycline bis-hydrochloride of  claim 14 , wherein in Step B-d) the storing is carried out for about 120 hours, and wherein in Step B-f) the storing is carried out for about 96 hours. 
     
     
         21 . Process for the preparation of crystalline eravacycline bis-hydrochloride of  claim 14 , wherein in step B-h) the solid is isolated by filtration, and washed with acetone, followed by a drying step, preferably wherein the drying step is carried out by leaving the washed isolated solid at room temperature for 3 hours. 
     
     
         22 . Process for the preparation of crystalline eravacycline bis-hydrochloride of formula III of  claim 14 , wherein in Step C-e) the acetone is added over a period of about 6 hours, and in step C-f) the suspension is stirred for about 24 hours at room temperature. 
     
     
         23 . Process for the preparation of crystalline eravacycline bis-hydrochloride of formula III of  claim 14 , wherein in step C-g) the solid is isolated by filtration, followed by a drying step. 
     
     
         24 . Process for the preparation of crystalline eravacycline bis-hydrochloride of formula III of  claim 23 , wherein the drying step is carried out at room temperature under vacuum. 
     
     
         25 . Process for the preparation of crystalline eravacycline bis-hydrochloride of formula III of  claim 24 , wherein the drying step is carried out at a vacuum of 30 mbar for about 20 hours. 
     
     
         26 . Process for the preparation of crystalline eravacycline bis-hydrochloride of formula III of  claim 14 , wherein the amount of seed crystals applied in step (ii) is in the range of from 1 to 10 weight %, preferably 2 to 5 weight %, based on the amount of eravacycline-bis-hydrochloride present in the aqueous solution provided in step (i). 
     
     
         27 . Process for the preparation of crystalline eravacycline bis-hydrochloride of formula III of  claim 14 , wherein the crystalline eravacycline bis-hydrochloride or the seed crystal is characterized by having a powder X-ray diffractogram comprising reflections at 2-theta angles of (5.6±0.2)°, (6.0±0.2)°, (7.1±0.2)°, (7.6±0.2)° and (8.6±0.2)°, when measured with Kalpha 1,2  radiation having a wavelength of 0.15419 nm and a temperature in the range of from 20 to 30° C. 
     
     
         28 . Process for the preparation of crystalline eravacycline bis-hydrochloride of formula III of  claim 14 , wherein the crystalline eravacycline bis-hydrochloride or the seed crystal is a non-stoichiometric hydrate. 
     
     
         29 . Process for the preparation of crystalline eravacycline bis-hydrochloride of formula III of  claim 14 , wherein the crystalline eravacycline bis-hydrochloride or the seed crystal is characterized by a water content in the range of from 8.0% to 12.5%, the crystalline eravacycline bis-hydrochloride being equilibrated at a relative humidity of 30% and 25.0±0.1° C., as determined by Karl-Fischer-Coulometry. 
     
     
         30 . Crystalline eravacycline bis-hydrochloride of formula (III) 
       
         
           
           
               
               
           
         
         or a solvate or a hydrate thereof, 
         wherein n is in the range from 1.8 to 2.2, and 
         wherein m is in the range from about 0.0 to 9.0, 
         wherein the crystalline eravacycline bis-hydrochloride is obtained by using seed crystals, 
         wherein said seed crystals are prepared by applying the process as defined in  claim 14 . 
       
     
     
         31 . The crystalline eravacycline bis-hydrochloride of  claim 30 , wherein the crystalline eravacycline bis-hydrochloride is a non-stoichiometric hydrate. 
     
     
         32 . The crystalline eravacycline bis-hydrochloride of  claim 30 , characterized by having a powder X-ray diffractogram comprising reflections at 2-theta angles of (5.6±0.2)°, (6.0±0.2)°, (7.1±0.2)°, (7.6±0.2)° and (8.6±0.2)°, when measured with Cu-Kalpha 1,2  radiation having a wavelength of 0.15419 nm and a temperature in the range of from 20 to 30° C. 
     
     
         33 . The crystalline eravacycline bis-hydrochloride of  claim 30 , characterized by a water content in the range of from 8.0% to 12.5%, the crystalline eravacycline bis-hydrochloride being equilibrated at a relative humidity of 30% and 25.0±0.1° C., as determined by Karl-Fischer-Coulometry. 
     
     
         34 . A pharmaceutical composition comprising an effective amount of crystalline eravacycline bis-hydrochloride as defined in  claim 30  and at least one pharmaceutically acceptable excipient. 
     
     
         35 . The pharmaceutical composition of  claim 34  which is an oral solid dosage form. 
     
     
         36 . The pharmaceutical composition of  claim 35  which is a tablet or a capsule. 
     
     
         37 . Process for the preparation of a pharmaceutical composition comprising eravacycline, wherein the pharmaceutical composition is intended for parenteral use, comprising the steps of:
 (i) providing the crystalline eravacycline bis-hydrochloride as defined in  claim 30  and optionally at least one stabilizer selected from the group of sugars and/or sugar alcohols;   (ii) dissolving or suspending crystalline eravacycline bis-hydrochloride and optionally the at least one stabilizer provided in step (i) in a solvent comprising water;   (iii) adjusting the pH of the solution or suspension obtained in step (ii) by adding at least one acid and/or base;   (iv) optionally filtering the solution or suspension obtained in step (iii) and   (v) lyophilizing the solution or suspension obtained in any one of steps (ii) to (iv) to give a pharmaceutical composition comprising eravacycline.   
     
     
         38 . A method for the treatment of bacterial infections, comprising administering an effective dosage of the pharmaceutical composition of  claim 34 .