Freeze-dried preparation containing high-purity pth and method for producing same

Abstract

[Problem] Provided is a freeze-dried preparation containing high-purity PTH peptide and a method for the production thereof. Also provided is a test method for PTH analogs to confirm the purity of a freeze-dried preparation containing PTH peptide, and the like. [Solution] In the present invention, the presence of PTH analogs produced during the manufacturing process of a freeze-dried preparation containing PTH peptide was confirmed. The production of these PTH analogs was also discovered to be markedly prevented or reduced by controlling exposure of the solution containing PTH peptide and the like to air environments within a pharmaceutical production facility.

1 . A freeze-dried preparation containing high-purity PTH peptide as an active ingredient, wherein “high-purity” means at least that the amount of at least one PTH analog versus the sum of the amount of PTH peptide and the total amount of PTH analogs in the preparation is 1.0% or less and/or that the total amount of PTH analogs versus the sum of the amount of PTH peptide and the total amount of PTH analogs is 5.0% or less; the freeze-dried preparation containing PTH peptide being produced by a method characterized in that the exposure of the solution containing PTH peptide prior to freeze drying to air environments within a pharmaceutical production facility is controlled. 
     
     
         2 . The freeze-dried preparation containing PTH peptide set forth in  claim 1  wherein the PTH analog is at least one or more among
 1) analog 1: 
 oxide of PTH peptide having a mass number 64 Da larger than the mass number of the PTH peptide contained in the preparation and producing digestion products corresponding to the following fragments (1-a) to (1-c) when the analog is digested by trypsin, 
 (1-a) Mass number of Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys (SEQ ID NO: 1)+16 Da, 
 (1-b) Mass number of His-Leu-Asn-Ser-Met-Glu-Arg (SEQ ID NO: 2)+16 Da, and 
 (1-c) Mass number of Val-Glu-Trp-Leu-Arg (SEQ ID NO: 3)+4 Da; 
 2) analog 2: 
 oxide of PTH peptide having a mass number 36 Da larger than the mass number of the PTH peptide contained in the preparation and producing digestion products corresponding to the following fragments (2-a) to (2-c) when the analog is digested by trypsin, 
 (2-a) Mass number of Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys (SEQ ID NO: 1)+16 Da, 
 (2-b) Mass number of His-Leu-Asn-Ser-Met-Glu-Arg (SEQ ID NO: 2)+16 Da, and 
 (2-c) Mass number of Val-Glu-Trp-Leu-Arg (SEQ ID NO: 3)+4 Da; 
 3) analog 3: 
 oxide of PTH peptide having a mass number 32 Da larger than the mass number of the PTH peptide contained in the preparation and producing digestion products corresponding to the following fragments (3-a) and (3-b) when the analog is digested by trypsin, 
 (3-a) Mass number of Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys (SEQ ID NO: 1)+16 Da and 
 (3-b) Mass number of His-Leu-Asn-Ser-Met-Glu-Arg (SEQ ID NO: 2)+16 Da; 
 4) analog 4: 
 oxide of PTH peptide having a mass number 48 Da larger than the mass number of the PTH peptide contained in the preparation and producing digestion products corresponding to the following fragments (4-a) and (4-b) when the analog is digested by trypsin, 
 (4-a) Mass number of Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys (SEQ ID NO: 1)+16 Da, 
 (4-b) Mass number of Val-Glu-Trp-Leu-Arg (SEQ ID NO: 3)+4 Da; 
 5) analog 5: 
 oxide of PTH peptide having a mass number 48 Da larger than the mass number of the PTH peptide contained in the preparation and producing digestion products corresponding to the following fragments (5-a) and (5-b) when the analog is digested by trypsin, 
 (5-a) Mass number of His-Leu-Asn-Ser-Met-Glu-Arg (SEQ ID NO: 2)+16 Da and 
 (5-b) Mass number of Val-Glu-Trp-Leu-Arg (SEQ ID NO: 3)+4 Da; 
 6) analog 6: 
 oxide of PTH peptide having a mass number 20 Da larger than the mass number of the PTH peptide contained in the preparation and producing digestion products corresponding to the following fragments (6-a) and (6-b) when the analog is digested by trypsin, 
 (6-a) Mass number of His-Leu-Asn-Ser-Met-Glu-Arg (SEQ ID NO: 2)+16 Da and 
 (6-b) Mass number of Val-Glu-Trp-Leu-Arg (SEQ ID NO: 3)+4 Da; 
 7) analog 7: 
 oxide of PTH peptide having a mass number 16 Da larger than the mass number of the PTH peptide contained in the preparation and producing a digestion product corresponding to the following fragment (7-a) when the analog is digested by trypsin, 
 (7-a) Mass number of Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys (SEQ ID NO: 1)+16 Da; 
 8) analog 8: 
 oxide of PTH peptide having a mass number 16 Da larger than the mass number of the PTH peptide contained in the preparation and producing a digestion product corresponding to the following fragment (8-a) when the analog is digested by trypsin, 
 (8-a) Mass number of His-Leu-Asn-Ser-Met-Glu-Arg (SEQ ID NO: 2)+16 Da; 
 9) analog 9: 
 oxide of PTH peptide having a mass number 32 Da larger than the mass number of the PTH peptide contained in the preparation and producing a digestion product corresponding to the following fragment (9-a) when the analog is digested by trypsin, 
 (9-a) Mass number of Val-Glu-Trp-Leu-Arg (SEQ ID NO: 3)+4 Da; 
 10) analog 10: 
 oxide of PTH peptide having a mass number 16 Da larger than the mass number of the PTH peptide contained in the preparation and producing a digestion product corresponding to the following fragment (10-a) when the analog is digested by trypsin, 
 (10-a) Mass number of Val-Glu-Trp-Leu-Arg (SEQ ID NO: 3)+16 Da; or 
 11) analog 11: 
 oxide of PTH peptide having a mass number 4 Da larger than the mass number of the PTH peptide contained in the preparation and producing a digestion product corresponding to the following fragment (11-a) when the analog is digested by trypsin, 
 (11-a) Mass number of Val-Glu-Trp-Leu-Arg (SEQ ID NO: 3)+4 Da. 
 
     
     
         3 . A freeze-dried preparation containing PTH peptide set forth in  claim 1  wherein the PTH analog is at least one or more among
 1) analog 1′: 
 oxide of PTH peptide in which residues corresponding to the position 8 and position 18 methionine of human PTH (1-34) have been changed into methionine sulfoxide residues and the residue corresponding to the position 23 tryptophan has been changed into a residue shown by the following structural formula (a); 
 
       
         
           
           
               
               
           
         
         2) analog 2′: 
         oxide of PTH peptide in which residues corresponding to the position 8 and position 18 methionine of human PTH (1-34) have been changed into methionine sulfoxide residues and the residue corresponding to the position 23 tryptophan has been changed into a residue shown by the following structural formula (b); 
       
       
         
           
           
               
               
           
         
         3) analog 3′: 
         oxide of PTH peptide in which residues corresponding to the position 8 and position 18 methionine of human PTH (1-34) have been changed into methionine sulfoxide residues; 
         4) analog 4′: 
         oxide of PTH peptide in which the residue corresponding to the position 8 methionine of human PTH (1-34) has been changed into a methionine sulfoxide residue and the residue corresponding to the position 23 tryptophan has been changed into a residue shown by the above structural formula (a); 
         5) analog 5′: 
         oxide of PTH peptide in which the residue corresponding to the position 18 methionine of human PTH (1-34) has been changed into a methionine sulfoxide residue and the residue corresponding to the position 23 tryptophan has been changed into a residue shown by the above structural formula (a); 
         6) analog 6′: 
         oxide of PTH peptide in which the residue corresponding to the position 18 methionine of human PTH (1-34) has been changed into a methionine sulfoxide residue and the residue corresponding to the position 23 tryptophan has been changed into a residue shown by the above structural formula (b); 
         7) analog 7′: 
         oxide of PTH peptide in which the residue corresponding to the position 8 methionine of human PTH (1-34) has been changed into a methionine sulfoxide residue; 
         8) analog 8′: 
         oxide of PTH peptide in which the residue corresponding to the position 18 methionine of human PTH (1-34) has been changed into a methionine sulfoxide residue; 
         9) analog 9′: 
         oxide of PTH peptide in which the residue corresponding to the position 23 tryptophan of human PTH (1-34) has been changed into a residue shown by the above structural formula (a); 
         10) analog 10′: 
         oxide of PTH peptide in which the residue corresponding to the position 23 tryptophan of human PTH (1-34) has been changed into a tryptophan monoxide residue shown by the following structural formula (c-1) or (c-2); 
       
       
         
           
           
               
               
           
         
       
       or
 11) analog 11′: 
 oxide of PTH peptide in which the residue corresponding to the position 23 tryptophan of human PTH (1-34) has been changed into a residue shown by the above structural formula (b). 
 
     
     
         4 . The freeze-dried preparation containing PTH peptide set forth in  claim 2  wherein high-purity means that the amount of at least one of the above analogs 1 to 11 versus the sum of the amount of PTH peptide and the total amount of PTH analogs in the preparation is 1.0% or less and/or that the total amount of the above analogs 1 to 11 versus the sum of the amount of PTH peptide and the total amount of PTH analogs is 5.0% or less. 
     
     
         5 . The freeze-dried preparation containing PTH peptide set forth in  claim 3  wherein high-purity means that the amount of at least one of the above analogs 1′ to 11′ versus the sum of the amount of PTH peptide and the total amount of PTH analogs in the preparation is 1.0% or less and/or that the total amount of the above analogs 1′ to 11′ versus the sum of the amount of PTH peptide and the total amount of PTH analogs is 5.0% or less. 
     
     
         6 . The freeze-dried preparation containing PTH peptide set forth in  claim 1  wherein the PTH peptide is human PTH (1-34). 
     
     
         7 . The freeze-dried preparation containing PTH peptide set forth in  claim 1  wherein the freeze-dried preparation containing PTH peptide is housed in a glass vial. 
     
     
         8 . The freeze-dried preparation containing PTH peptide set forth in  claim 1 , characterized in that exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying is controlled in any one or more steps selected from a step for preparing a solution containing PTH peptide, an aseptic filtration step, a drug solution dispensing step, and a step for loading into a freeze drying means. 
     
     
         9 . The freeze-dried preparation containing PTH peptide set forth in  claim 8  characterized in being produced using a method that also includes control of exposure of the freeze-dried product to air environments within a pharmaceutical production facility in a vial sealing step after freeze drying. 
     
     
         10 . The freeze-dried preparation containing PTH peptide set forth in  claim 1  characterized in that exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying is controlled in the step for loading into the freeze drying means. 
     
     
         11 . The freeze-dried preparation containing PTH peptide set forth in  claim 10  characterized in that the above exposure is controlled by using a freeze-drying chamber equipped with means for controlling the entrance of air within a pharmaceutical production facility into the freeze-drying means. 
     
     
         12 . The freeze-dried preparation containing PTH peptide set forth in  claim 11  characterized in that the freeze-drying means is a freeze-drying chamber having an easily openable and closable sub-door provided in an opening created in a small door unit opened when containers housing the solution containing PTH peptide prior to freeze drying are loaded into and unloaded from this means, thereby controlling exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying by opening this sub-door only during container loading and quickly closing the sub-door after loading. 
     
     
         13 . The freeze-dried preparation containing PTH peptide set forth in  claim 11  wherein the freeze-drying means is a freeze-drying chamber having an opening created in a small door unit opened when containers housing the solution containing PTH peptide prior to freeze drying are loaded into and unloaded from this means, and the means for controlling the ingress of air within a pharmaceutical production facility into the freeze-drying means is an airflow-adjusting cover that can change the air flow to a direction not directed from this opening to the inside of the chamber. 
     
     
         14 . The freeze-dried preparation containing PTH peptide set forth in  claim 10  characterized in that the loading step controls exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying by purging the inside of the freeze-drying means with an inert gas. 
     
     
         15 . The freeze-dried preparation containing PTH peptide set forth in  claim 10  wherein the freeze-drying means is a freeze-drying chamber having an easily openable and closable sub-door provided in an opening created in a small door unit opened when containers housing the solution containing PTH peptide prior to freeze drying are loaded into and unloaded from this means, thereby controlling exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying by opening this sub-door only during container loading and quickly closing the sub-door after loading and purging the inside of the freeze-drying means with an inert gas in the loading step. 
     
     
         16 . The freeze-dried preparation containing PTH peptide set forth in  claim 10  wherein the freeze-drying means is a freeze-drying chamber having an opening created in a small door unit opened when containers housing the solution containing PTH peptide prior to freeze drying are loaded into and unloaded from this means, this opening being equipped with an airflow-adjusting cover, thereby controlling exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying by changing the direction of the airflow-adjusting cover so that air is not directed into the chamber and purging the inside of the freeze-drying means with an inert gas in the loading step. 
     
     
         17 . The freeze-dried preparation containing PTH peptide set forth in  claim 10  wherein the loading step is a step that spans three or more hours. 
     
     
         18 . The freeze-dried preparation containing PTH peptide set forth in  claim 8  wherein time from the beginning of the step for preparing a solution containing PTH peptide to the end of the step for loading into the freeze-drying means spans three or more hours, and production is performed using a method for controlling exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility in one or more steps during the time. 
     
     
         19 . The freeze-dried preparation containing PTH peptide set forth in  claim 14  wherein the inert gas is nitrogen gas. 
     
     
         20 . A freeze-dried preparation containing high-purity PTH peptide as an active ingredient, the PTH-peptide-containing freeze-dried preparation manufactured using a method characterized in that exposure of a solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying is controlled during loading into the freeze-drying means; wherein “high-purity” means at least that the amount of at least one PTH analog versus the sum of the amount of PTH peptide and the total amount of PTH analogs in the preparation is 1.0% or less and/or that the total amount of PTH analogs versus the sum of the amount of PTH peptide and the total amount of PTH analogs is 5.0% or less; the loading step is a step that spans three or more hours; the air environment is an environment that maintains one-way air flow of clean air that has passed through an HEPA filter downward from above; and the velocity of the air flow 20 cm directly under the HEPA filter is 0.2-1.0 m/s. 
     
     
         21 . A method for producing a freeze-dried preparation containing PTH peptide, the method being characterized in that exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility is controlled in one or more steps from the beginning of the step for preparing a solution containing PTH peptide to the end of the step for loading into the freeze-drying means. 
     
     
         22 . The method set forth in  claim 21  wherein exposure of the freeze-dried product to air environments within a pharmaceutical production facility is also controlled in the step for sealing vials after freeze drying. 
     
     
         23 . The method set forth in  claim 21  characterized in that exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility is controlled in the step for loading into the freeze-drying means. 
     
     
         24 . The method set forth in  claim 23  characterized in that exposure is controlled using a freeze-drying chamber equipped with means for controlling the inflow of air within a pharmaceutical production facility into the freeze-drying means. 
     
     
         25 . The method set forth in  claim 23  wherein the step for loading into the freeze-drying means is a step that spans three or more hours. 
     
     
         26 . The method set forth in  claim 24  wherein the freeze-drying means is a freeze-drying chamber having an easily openable and closable sub-door provided in an opening created in a small door unit opened when containers housing the solution containing PTH peptide prior to freeze drying are loaded into and unloaded from this means, thereby controlling exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying by opening this sub-door only during container loading and quickly closing the sub-door after loading. 
     
     
         27 . The method set forth in  claim 24  wherein the freeze-drying means is a freeze-drying chamber having an opening created in a small door unit opened when containers housing the solution containing PTH peptide prior to freeze drying are loaded into and unloaded from this means, and the means for controlling the ingress of air within a pharmaceutical production facility into the freeze-drying means is an airflow-adjusting cover that can change the air flow to a direction not directed from this opening to the inside of the chamber. 
     
     
         28 . The method set forth in  claim 23  characterized in that exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying is controlled by purging the inside of the freeze-drying means with an inert gas. 
     
     
         29 . The method set forth in  claim 23  characterized in that the freeze-drying means is a freeze-drying chamber having an easily openable and closable sub-door provided in an opening created in a small door unit opened when containers housing the solution containing PTH peptide prior to freeze drying are loaded and unloaded, thereby controlling exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying by opening this sub-door only during container loading and quickly closing the sub-door after loading and purging the inside of the freeze-drying means with an inert gas in the loading step. 
     
     
         30 . The method set forth in  claim 23 , characterized in that the freeze-drying means is a freeze-drying chamber having an opening created in a small door unit opened when containers housing the solution containing PTH peptide prior to freeze drying are loaded into and unloaded from this means, the opening being equipped with an airflow-adjusting cover, thereby controlling exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying by changing the airflow-adjusting cover to a direction in which the air flow is not directed into the chamber and purging the inside of the freeze-drying means with an inert gas in the loading step. 
     
     
         31 . The method set forth in  claim 28  wherein the inert gas is nitrogen. 
     
     
         32 . The method set forth in  claim 21  wherein the container housing the solution containing PTH peptide is a glass vial. 
     
     
         33 . The method set forth in  claim 21  wherein the PTH is human PTH (1-34). 
     
     
         34 . The method set forth in  claim 21  wherein the air environment within a pharmaceutical production facility is an air environment in which 1) the air is of grade A, 2) clean air that has passed through an HEPA filter having the ability to trap particles having a particle size of 0.3 μm at an efficiency of 99.97% or higher is maintained as a one-way air flow downward from above, and 3) the ozone concentration is 0.001-0.1 ppm. 
     
     
         35 . The method set forth in  claim 21  wherein the air environment within a pharmaceutical production facility is an air environment containing a formaldehyde concentration of 0.1 ppm or less. 
     
     
         36 . The method set forth in  claim 21  wherein the amount of at least one PTH analog versus the sum of the amount of PTH peptide and total amount of PTH analogs is 1.0% or less and/or the total amount of PTH analogs versus the sum of the amount of PTH peptide and total amount of PTH analogs is 5.0% or less in the freeze-dried preparation containing PTH peptide. 
     
     
         37 . A method for producing a freeze-dried preparation containing PTH peptide, the method being characterized in that exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility is controlled in one or more steps from the beginning of the step for preparing a solution containing PTH peptide to the end of the step for loading into the freeze-drying means, for controlling the production of PTH analogs 1 to 11 set forth in  claim 2 . 
     
     
         38 . A method for producing a freeze-dried preparation containing PTH peptide, the method being characterized in that exposure of the solution containing PTH peptide to air environments within a pharmaceutical production facility is controlled in one or more steps from the beginning of the step for preparing a solution containing PTH peptide to the end of the step for loading into the freeze-drying means, to control the production of PTH analogs to 1′ 11′ set forth in  claim 3 . 
     
     
         39 . A freeze-dried preparation containing PTH peptide manufactured using the method set forth in  claim 21 . 
     
     
         40 . A method for producing a freeze-dried preparation containing high-purity PTH peptide as an active ingredient, the method characterized in that exposure of a solution containing PTH peptide to air environments within a pharmaceutical production facility prior to freeze drying is controlled during loading into the freeze-drying means; wherein “high-purity” means at least that the amount of at least one PTH analog versus the sum of the amount of PTH peptide and the total amount of PTH analogs is 1.0% or less and/or that the total amount of PTH analogs versus the sum of the amount of PTH peptide and the total amount of PTH analogs is 5.0% or less in the preparation; the loading step is a step that spans three or more hours; the air environment is an environment in which clean air that has passed through an HEPA filter is maintained as a one-way air flow downward from above; and the velocity of the air flow 20 cm directly under the HEPA filter is 0.2-1.0 m/s. 
     
     
         41 . A method for testing a freeze-dried preparation containing PTH peptide, the method being characterized in confirming the presence of at least one or more of the PTH analogs 1 to 11 of  claim 2  and/or determining the amounts present in the freeze-dried preparation containing PTH peptide. 
     
     
         42 . A method for testing a freeze-dried preparation containing PTH peptide, the method being characterized in confirming the presence of at least one or more of the PTH analogs 1′ to 11′ of  claim 3  and/or determining the amounts present in the freeze-dried preparation containing PTH peptide. 
     
     
         43 . The method set forth in  claim 41  wherein determination of the PTH analogs includes calculating the area of the peak corresponding to the PTH analog on a chromatogram when the ultraviolet absorption of a sample derived from a freeze-dried preparation containing PTH peptide is measured by high-performance liquid chromatography. 
     
     
         44 . The method set forth in  claim 43  including calculation of the purity of the PTH peptide in the freeze-dried preparation containing PTH peptide by comparing the area of a peak corresponding to a PTH analog on a chromatogram and the peak area corresponding to PTH peptide or the sum of the peak area of PTH peptide and the peak area of all other PTH analogs detected on the same chromatogram when the ultraviolet absorbance of a sample derived from a freeze-dried preparation containing PTH peptide is measured by high-performance liquid chromatography. 
     
     
         45 . The method set forth in  claim 44  including calculation of the purity of the PTH peptide in a freeze-dried preparation containing PTH peptide by comparing the area of that peak and the peak area corresponding to PTH peptide or the sum of the peak area of PTH peptide and the peak area of all other PTH analogs detected on the same chromatogram when using chromatography conditions such that any two or more PTH analogs are detected as one or more single peaks on the chromatogram. 
     
     
         46 . The method set forth in  claim 41  for ensuring that the amount of at least one PTH analog versus the sum of the amount of PTH peptide and the total amount of PTH analogs is 1.0% or less and/or the total amount of PTH analogs versus the sum of the amount of PTH peptide and the total amount of PTH analogs is 5.0% or less in a freeze-dried preparation containing PTH peptide. 
     
     
         47 . The method set forth in  claim 41  including the detection of the mass number of the PTH analogs using a high-performance liquid chromatograph-mass spectrometer. 
     
     
         48 . The method set forth in  claim 41  including fractionating a substance that gives a single peak on the chromatogram and identifying the mass number of the fragments produced by digesting this substance using trypsin. 
     
     
         49 . A method for producing a pharmaceutical comprising a freeze-dried preparation containing PTH peptide including a step to carry out the test method of  claim 41 .