Control method for removing acetonitrile in low-temperature polypeptide medicine

Abstract

The invention relates to a method for removing and controlling acetonitrile in low-temperature polypeptide medicines, which comprises the following steps of: removing acetonitrile in a refined peptide aqueous solution to a control range A at a low temperature of 10-20 DEG C from a polypeptide raw material medicine which contains acetonitrile solvent residue and has the mass volume concentration of 10-50g / L, namely the refined peptide aqueous solution, and then removing the acetonitrile by a primary freeze-drying method, the acetonitrile content in the freeze-dried peptide bulk drug is qualified; wherein A is liquid acetonitrile residue detected by a gas phase method, and A is less than or equal to 410 * E / C / 1000. According to the method, the acetonitrile can be quickly removed at low temperature by adopting efficient low-temperature concentration equipment, and the acetonitrile residue can be ensured to be qualified after freeze-drying by cooperating with gas phase detection and process control, so that the method is suitable for mass production of products.

Description

Technical Field

[0001] This invention relates to a method for removing organic solvents, particularly a method for controlling the removal of acetonitrile in low-temperature polypeptide drugs. Background Technology

[0002] Peptide drugs are heat-sensitive, and high temperatures can lead to a significant increase in hydrolysis and racemic impurities. Their purification process requires the use of acetonitrile as the organic phase in chromatographic separation, and subsequent purification makes it difficult to remove trace amounts of acetonitrile from the liquid. Furthermore, the pharmacopoeia requires that acetonitrile residues not exceed 410 ppm.

[0003] Currently, the industry has methods such as salting out and freeze-drying, but due to limitations in product properties and equipment capacity, it has not yet been able to achieve large-scale production.

[0004] The method of concentration and freeze-drying depends on the configuration of the concentration equipment and freeze dryer. For small-batch production, a glass rotary evaporator concentration followed by one freeze-drying process can be used. As production increases, the limits of the concentration equipment may prevent the acetonitrile residue from being qualified in one freeze-drying process, so two freeze-drying processes are required. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to address the shortcomings of the prior art by providing a method for controlling the removal of acetonitrile in low-temperature polypeptide drugs. This method uses a high-efficiency low-temperature concentration device to quickly remove acetonitrile at low temperatures, and combines it with gas phase detection to control acetonitrile residues during the process, ensuring that the acetonitrile residues in the freeze-dried product are within acceptable limits.

[0006] The technical problem to be solved by this invention is achieved through the following technical solution. This invention is a method for controlling the removal of acetonitrile in low-temperature polypeptide drugs, characterized by:

[0007] The technical problem to be solved by this invention can also be further achieved through the following technical solutions. The acetonitrile removal and control method for low-temperature polypeptide drugs described above is characterized by: a polypeptide raw material (i.e., an aqueous solution containing acetonitrile solvent residue at a mass-volume concentration of 10-50 g / L) is subjected to low-temperature treatment (10℃-20℃) to remove acetonitrile from the aqueous solution to a control range A, followed by a single freeze-drying process to remove acetonitrile completely, achieving a qualified acetonitrile content in the freeze-dried polypeptide raw material; wherein A is the liquid acetonitrile residue detected by a gas phase method, in ppm; and A≤410×E / C / 1000;

[0008] In the formula:

[0009] A represents the residual acetonitrile in liquid as measured by a gas chromatography method (ppm); [The result was obtained from the detection].

[0010] E represents the concentration of the peptide, in g / L; the concentration is between 10 and 50 g / L; it is obtained through detection; C represents the acetonitrile removal residual rate threshold during the freeze-drying process, and C = 1%-5%.

[0011] A further preferred embodiment of the method for controlling acetonitrile removal in low-temperature polypeptide drugs according to the present invention comprises the following specific steps:

[0012] (1) Set the acetonitrile removal residual rate threshold for the freeze-drying process to C, and C = 1% - 5%;

[0013] (2) A linear curve was prepared by using gas chromatography to determine the acetonitrile content A in the aqueous solution of the peptide;

[0014] (3) The formula was used to estimate the residual acetonitrile B2 in the solid after the lyophilization of the peptide;

[0015] (4) Based on the final requirement that the acetonitrile residue B2 is less than 410 ppm, the upper limit of the acetonitrile residue in the previous step of the peptide aqueous solution is calculated in reverse, which is the upper limit of A.

[0016] (5) The product is processed by freeze drying and tested until the acetonitrile residue rate is qualified.

[0017] Once liquid acetonitrile residue A is confirmed, the solid acetonitrile content B2 after freeze-drying can be calculated using the following formula: A≤410×E / C / 1000;

[0018] D1 = F × A / 1000;

[0019] D2 = D1 × C;

[0020] B2 = D2 / (E × F) * 1000000;

[0021] In the formula:

[0022] F represents the volume of the sperm peptide, in L; this was detected.

[0023] D1 represents the amount of acetonitrile in the peptide, in grams; this was determined by detection.

[0024] Acetonitrile content in D2 lyophilized peptide, in g;

[0025] B2 is the residual solid acetonitrile value in the lyophilized peptide. B2 is less than 410 ppm.

[0026] A further preferred technical solution of the method for controlling the removal of acetonitrile in a low-temperature polypeptide drug according to the present invention is that when the residual acetonitrile in the liquid (ppm, i.e., the A value) meets the requirements, the A value is reduced by distillation.

[0027] A further preferred embodiment of the method for controlling the removal of acetonitrile in a low-temperature polypeptide drug according to the present invention is that when the concentration of E (acetonitrile) does not meet the requirement of 10-50 g / L, its concentration is increased or decreased by adding water or by distillation.

[0028] Compared with the prior art, the present invention has the following beneficial effects: The acetonitrile removal and control method in the low-temperature polypeptide drugs of the present invention uses a high-efficiency low-temperature concentration device to quickly remove acetonitrile at low temperature. Combined with gas phase detection, the process control of acetonitrile residue can ensure that the acetonitrile residue of the finished product after freeze-drying is qualified. Detailed Implementation

[0029] The specific technical solutions of the present invention are further described below to enable those skilled in the art to further understand the present invention, without constituting a limitation on its rights.

[0030] Example 1: A method for controlling acetonitrile removal in a low-temperature peptide drug. The method involves treating a peptide raw material (i.e., an aqueous solution containing acetonitrile solvent residue at a concentration of 30 g / L) at 15°C. The acetonitrile in the aqueous solution is then removed to a control range A, followed by a single lyophilization process to remove all remaining acetonitrile, achieving a qualified acetonitrile content in the lyophilized peptide raw material. Here, A represents the residual liquid acetonitrile detected by a gas chromatography method, in ppm, and A ≤ 410 × E / C / 1000.

[0031] In the formula:

[0032] A represents the residual acetonitrile in liquid as measured by a gas chromatography method (ppm); [The result was obtained from the detection].

[0033] E represents the concentration of the peptide, in g / L; the concentration is between 10 and 50 g / L; it is obtained through detection; C represents the acetonitrile removal residual rate threshold during the freeze-drying process, and C = 1%-5%.

[0034] The specific steps are as follows:

[0035] (1) Set the acetonitrile removal residual rate threshold for the freeze-drying process to C, and C = 1% - 5%;

[0036] (2) A linear curve was prepared by using gas chromatography to determine the acetonitrile content A in the aqueous solution of the peptide;

[0037] (3) The formula was used to estimate the residual acetonitrile B2 in the solid after the lyophilization of the peptide;

[0038] (4) Based on the final requirement that the acetonitrile residue B2 is less than 410 ppm, the upper limit of the acetonitrile residue in the previous step of the peptide aqueous solution is calculated in reverse, which is the upper limit of A.

[0039] (5) The product is processed by freeze drying and tested until the acetonitrile residue rate is qualified.

[0040] Once liquid acetonitrile residue A is confirmed, the solid acetonitrile content B2 after freeze-drying can be calculated using the following formula: A≤410×E / C / 1000;

[0041] D1 = F × A / 1000;

[0042] D2 = D1 × C;

[0043] B2 = D2 / (E × F) * 1000000;

[0044] In the formula:

[0045] F represents the volume of the sperm peptide, in L; this was detected.

[0046] D1 represents the amount of acetonitrile in the peptide, in grams; this was determined by detection.

[0047] Acetonitrile content in D2 lyophilized peptide, in g;

[0048] B2 is the residual solid acetonitrile value in the lyophilized peptide. B2 is less than 410 ppm.

[0049] When the residual acetonitrile (ppm), i.e., value A, meets the requirements, distillation is used to reduce the value A. When the concentration of peptide E (E) does not meet the requirement of 30 g / L, water is added or distillation is used to increase or decrease its concentration.

[0050] Example 2, Acetonitrile Removal Control Method in Low-Temperature Peptide Drugs

[0051] 1.1 Experiments were conducted to determine the acetonitrile removal rate of the existing freeze-drying process;

[0052] 1.2 Develop a gas phase detection method, generate a linear curve, and calibrate the acetonitrile content in the concentrate;

[0053] 1.3 Calculation of acetonitrile residue in the solid after drying of the peptide;

[0054] 1.4 Based on the final requirement that the acetonitrile residue should not exceed 410 PPM, the upper limit of the acetonitrile residue in the concentrate from the previous step is calculated in reverse.

[0055] 1.5 After the freeze-drying process, the acetonitrile residue rate of the product passed the test.

[0056] The relationship values ​​are shown in the table below:

[0057]

[0058] In the table, A represents detection using a gas chromatography method;

[0059] Acetonitrile removal rate C varies depending on the equipment and freeze-drying process, with a normal range of 1%-5%.

[0060] Once liquid acetonitrile residue is confirmed, the solid acetonitrile content of the peptide can be calculated using the following formula:

[0061] The final result B2 should be less than 410 ppm;

[0062] A≤410×E / C / 1000;

[0063] D1 = F × A / 1000;

[0064] D2 = D1 × C;

[0065] B2 = D2 / (E×F)×1000000;

[0066] Then we can deduce that A≤D1 / F, A≤B2×E / C, and by substituting B2 into 410, A≤410×E / C.

[0067] The following are experiments conducted by the inventor:

[0068] Batch number: 24601

[0069]

[0070]

[0071] The acetonitrile content in the liquid was tested to be 351 ppm, and the acetonitrile content in the final product was 170 ppm. The lyophilized acetonitrile removal rate was 1.45% (by reverse calculation), and the acetonitrile content in the finished product met the requirements.

[0072] Batch number: 240701

[0073]

[0074] The acetonitrile content in the liquid was tested to be 381 ppm, and the acetonitrile content in the final product was 158.75 ppm. The lyophilized acetonitrile removal rate was 1.5% (by reverse calculation), and the acetonitrile content in the finished product met the requirements.

[0075] Batch number: 240801

[0076]

[0077] The acetonitrile content in the liquid was tested to be 500 ppm, and the acetonitrile content in the final product was 250 ppm. The lyophilized acetonitrile removal rate was 2% (by reverse calculation), and the acetonitrile content in the finished product met the requirements.

[0078] Based on the data summary from the previous three steps, an acetonitrile removal residual rate of 2% is within the safe range. At a freeze-drying concentration of 30g / L-40g / L in the workshop and with a full load, the acetonitrile content of the peptide can be calculated to be at the lower limit concentration A ≤ 615ppm. This enables large-scale batch production.

Claims

1. A method for controlling the removal of acetonitrile in a low-temperature polypeptide drug, characterized in that: A polypeptide raw material containing acetonitrile solvent residue at a concentration of 10-50 g / L (i.e., an aqueous solution of crystalline peptide) is subjected to low-temperature treatment (10℃-20℃) to remove acetonitrile to the control range A. Then, the acetonitrile is removed again by a single freeze-drying process to achieve the required acetonitrile content in the freeze-dried peptide raw material. Here, A represents the residual liquid acetonitrile detected by gas chromatography, in ppm, and A ≤ 410 × E / C / 1000. In the formula: A represents the residual acetonitrile in liquid as measured by a gas chromatography method (ppm); [The result was obtained from the detection]. E represents the concentration of the peptide, in g / L; the concentration is between 10 and 50 g / L; it is obtained by detection; C represents the acetonitrile removal residual rate threshold during the freeze-drying process, and C = 1%-5%.

2. The method for controlling acetonitrile removal in a low-temperature polypeptide drug according to claim 1, characterized in that, The specific steps are as follows: (1) Set the acetonitrile removal residual rate threshold for the freeze-drying process to C, and C = 1%-5%; (2) A linear curve was constructed using gas chromatography to calibrate the acetonitrile content A in the aqueous solution of the peptide; (3) The residual acetonitrile B2 in the lyophilized solid of the peptide was calculated using the formula; (4) Based on the final requirement that the acetonitrile residue B2 is less than 410 ppm, the upper limit of the acetonitrile residue in the previous step of the peptide aqueous solution is calculated in reverse, which is the upper limit of A. (5) The product is processed by freeze drying and tested until the acetonitrile residue rate is qualified; Once the residual liquid acetonitrile A is confirmed, the content of solid acetonitrile B2 after lyophilization can be calculated using the following formula: A≤410×E / C / 1000; D1 = F × A / 1000; D2 = D1 × C; B2 = D2 / (E × F) * 1000000; In the formula: F represents the volume of the sperm peptide, in L; this was detected. D1 represents the amount of acetonitrile in the peptide, in grams. Detected; Acetonitrile content in D2 lyophilized peptide, in g; B2 is the residual solid acetonitrile value in the lyophilized peptide. B2 is less than 410 ppm.

3. The method for controlling acetonitrile removal in a low-temperature polypeptide drug according to claim 1 or 2, characterized in that: When the residual acetonitrile in liquid (ppm), i.e., the A value, meets the requirements, the A value is reduced by distillation.

4. The method for controlling acetonitrile removal in a low-temperature polypeptide drug according to claim 1 or 2, characterized in that: When the concentration of E (the peptide) is not within the range of 10-50 g / L, its concentration can be increased or decreased by adding water or by distillation.

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