Method for pulverizing polypeptide drug

Abstract

A method for pulverizing a polypeptide drug, which method comprises the following steps: S1) lyophilization: obtaining a polypeptide drug lyophilized powder by means of lyophilization; S2) sieving: taking the polypeptide drug lyophilized powder, and subjecting same to a sieving treatment using a sieve mesh of 5-200 mesh to obtain a sieved polypeptide drug powder; and S3) jet milling: processing the sieved polypeptide drug powder using a jet mill to obtain a polypeptide drug powder, wherein the milling air pressure in the jet milling step is set to 1-10 bar. Compared with the freeze-dried samples in the prior art, the pulverized product obtained by means of pulverization exhibits more uniform characteristics, a smaller particle size, a narrower particle size distribution, and a higher intrinsic dissolution rate, and thus can meet the requirements of oral polypeptide solid preparations.

Description

A method for pulverizing a polypeptide drug TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical preparations, in particular to a method for pulverizing a polypeptide drug. BACKGROUND

[0002] Glucagon-like peptide-1 (GLP-1) is a hormone mainly secreted by intestinal cells, which belongs to the incretins. GLP-1 receptor agonists (GLP-1RAs) are a class of drugs for reducing blood glucose and weight that have emerged in recent years. The chemical nature of this class of drugs is polypeptide. By activating the GLP-1 receptor (GLP-1R), it enhances insulin secretion and suppresses glucagon secretion in a glucose concentration-dependent manner, and can reduce food intake by delaying gastric emptying, enhancing satiety, and suppressing appetite, thereby achieving the effects of reducing blood glucose and weight loss. Typical GLP-1RAs include liraglutide, dulaglutide, semaglutide, tirzepatide, etc. On the basis of the GLP-1R target, many pharmaceutical research institutions have further modified GLP-1RAs at the molecular level to make them become dual-target or triple-target drugs. For example: GLP-1 and GIP dual-target receptor agonists (GIPR / GLP-1RAs), a typical drug is tirzepatide from Eli Lilly; GLP-1 and GCGR dual-target receptor agonists (GCGR / GLP-1RAs), a typical drug is semaglutide developed by Eli Lilly and Sinotech; GLP-1 and AMY dual-target receptor agonists (AMY / GLP-1RAs), a typical drug is Amycretin from Novo Nordisk; GLP-1R / GIPR / GCGR triple-target receptor agonists, a typical drug is Retatrutide from Eli Lilly. Although polypeptide drugs have advantages such as low toxicity, low immunogenicity, and high tissue permeability, they also have disadvantages such as easy enzymatic hydrolysis, poor stability, and short half-life. According to the physicochemical properties of polypeptides, pharmaceutical research institutions continue to improve the molecular structure and preparation process of GLP-1RAs to improve their resistance to enzymes and prolong their half-life, achieve breakthroughs from daily preparations to weekly preparations, and improve the effects of reducing blood glucose and weight loss. It is worth noting that the current research and development of polypeptide GLP-1RAs, whether single-target or multi-target drugs, their main dosage form is injection. The only oral version of semaglutide (trade name Rybelsus) can achieve oral administration, while the oral versions of Amycretin from Novo Nordisk and VK-2735 from Viking Therapeutics are still in the clinical research stage. Compared with injection, oral administration has obvious advantages in patient compliance.

[0003] The oral delivery of polypeptide drugs is affected not only by the preparation process, but also by the process of the bulk drug. Because of the poor stability of polypeptide drugs, freeze-drying is often preferred as the drying method during the preparation of polypeptide bulk drugs. Although freeze-drying can avoid or reduce the degradation of polypeptide drugs to the greatest extent, the polypeptide bulk drugs obtained by this method are often in the form of loose blocks, flakes, needles or irregular shapes. For polypeptide injections, since the solid bulk drug needs to be finally dissolved into a liquid, the properties of the bulk drug have less impact on the preparation and drug efficacy. For polypeptide solid preparations, since it is administered orally, the irregular properties of the bulk drug often affect the uniformity of the mixing of the raw and auxiliary materials, the content uniformity, the granulation formability, etc., which affects the in vitro and in vivo dissolution behavior and bioavailability of polypeptide drugs, and ultimately affects the drug efficacy. Therefore, polypeptide bulk drugs obtained by freeze-drying are usually not suitable for the preparation of oral polypeptide solid preparations. In summary, for oral polypeptide solid preparations, special attention should be paid to the impact of the properties of polypeptide bulk drugs on the preparation and drug efficacy.

[0004] In order to obtain polypeptide bulk drugs with more uniform properties, Novo Nordisk prepared semaglutide tablet bulk drugs by spray drying (patent numbers 201980084815.3, 202080021713.X and 202080061445.4). Spray drying is a kind of thermal drying method. The feed liquid is atomized in the drying tower through an atomizer, forming tiny droplets and contacting with high-temperature gas. Because of the large evaporation area, solvent evaporation and drying can be achieved in a short time to obtain the finished product. Spray drying is efficient and can obtain polypeptide bulk drugs with uniform properties, small particle size and narrow particle size distribution. However, because high-temperature gas above 100°C is needed, sample degradation and purity reduction are inevitable during the drying process. For example, patent 202080021713.X introduces a spray drying process for GLP-1 peptide, and the purity of the sample will decrease by about 0.2-2.0% during the spray drying process. Qilu Pharmaceutical prepared semaglutide tablet bulk drugs by spray freeze-drying (patent number 202010911687.X). Spray freeze-drying is a kind of cold drying method. The feed liquid is atomized through an atomizer while contacting with low-temperature gas, instantaneously forming tiny ice crystal particles, and then freeze-drying to obtain the finished product. Spray freeze-drying can realize sample preparation at low temperature without causing sample degradation, and the obtained polypeptide bulk drugs have uniform properties. However, there is a problem of low recovery rate and large loss. For example, patent 202010911687.X introduces a spray freeze-drying method for preparing semaglutide microparticles, which can obtain spherical semaglutide bulk drugs with a particle size of 1-100 μm, a purity of >99.0%, and a single impurity of <0.10%, but the recovery rate is only 90-94%.

[0005] In addition to the above-mentioned spray drying and spray freeze drying methods, the most commonly used polypeptide bulk drug is freeze drying technology. Freeze drying technology has mild conditions, and the sample is not usually degraded during the drying process, and is particularly suitable for polypeptide drugs with heat sensitivity, but the properties of polypeptide freeze-dried powder are often uneven, in the form of loose blocks, sheets, needles or irregular shapes, etc. The particle size is large, and the particle size distribution is wide. Since the dosage form of polypeptide drugs is mainly injection, polypeptide freeze-dried powder needs to be reconstituted before it can be prepared into a preparation, so the above characteristics have little effect on polypeptide injection. Oral solid preparations have higher requirements for the properties of raw materials, and usually require uniform properties, small particle size and narrow particle size distribution of raw materials, so polypeptide freeze-dried powder cannot meet the needs of polypeptide oral solid preparations. SUMMARY

[0006] In view of the above technical deficiencies, the present application provides a multi-step crushing method for polypeptide samples, which comprises sieving, freeze drying and airflow crushing technology to crush the sample, and the obtained polypeptide bulk drug has uniform properties, small particle size, narrow particle size distribution, high purity, no degradation, high yield, and faster inherent dissolution rate, and is suitable for the preparation of oral polypeptide solid preparations.

[0007] The first aspect of the present application provides a crushing method for polypeptide drugs, which comprises the following steps:

[0008] S1) Freeze drying: obtaining polypeptide drug freeze-dried powder by freeze drying;

[0009] S2) Sieving: taking the polypeptide drug freeze-dried powder, sieving the polypeptide drug freeze-dried powder with a 5-200 mesh sieve to obtain sieved polypeptide drug powder;

[0010] S3) Airflow crushing: treating the sieved polypeptide drug powder with an airflow crusher to obtain polypeptide drug powder;

[0011] In the airflow crushing step, the crushing gas pressure is set to 1 bar-10 bar.

[0012] Further, the gas source used in the airflow crusher is compressed air or inert gas.

[0013] Further, the crushing gas pressure is set to 1 bar-5 bar, for example, 1 bar, 2 bar, 3 bar, 4 bar.

[0014] Further, the crushing gas pressure is set to 2.5 bar-3.5 bar.

[0015] Further, the crushing gas pressure is set to 3 bar.

[0016] Further, the sieve is 5 mesh, 10 mesh, 20 mesh, 50 mesh, 100 mesh or 200 mesh.

[0017] Further, the polypeptide drug is a GLP-1 analogue.

[0018] Still further, the GLP-1 analogue is selected from the group consisting of liraglutide, losepramide, dulaglutide, semaglutide, exenatide, albiglutide, medinadide, amycretin, rithaleride.

[0019] Further, the yield of the pulverization method is ≥ 95.0%. The calculation method of the yield is as follows:

[0020] Yield / % = (obtained polypeptide drug powder / polypeptide drug lyophilized powder) x 100%.

[0021] The second aspect of the present application provides a polypeptide drug powder obtained by the above-mentioned pulverization method, wherein the polypeptide drug powder has a purity ≥ 99.5% and a single impurity < 0.10%.

[0022] Further, the particle size distribution of the polypeptide drug is D10 of 0.5 μm-6 μm, D50 of 2 μm-22 μm, and D90 of 4 μm-35 μm.

[0023] Further, the particle size distribution of the polypeptide drug is D10 of 1 μm-3 μm, D50 of 8 μm-15 μm, and D90 of 15 μm-28 μm.

[0024] Further, the inherent dissolution rate of the air flow pulverized polypeptide drug is significantly faster than that of the lyophilized polypeptide drug.

[0025] The third aspect of the present application provides the use of the above-mentioned polypeptide drug powder as a bulk drug in the preparation of an oral polypeptide solid drug. Advantages

[0026] The polypeptide drug powder prepared by the present application has uniform properties, small particle size, narrow particle size distribution, high recovery rate, high purity, and faster inherent dissolution rate, which can meet the needs of oral polypeptide solid preparations. Specifically, compared with the freeze-dried sample in the prior art, the pulverized product of the present application has more uniform properties, smaller particle size, narrower particle size distribution, and faster inherent dissolution rate, which can meet the needs of oral polypeptide solid preparations; compared with the spray-dried sample in the prior art, the pulverized product of the present application has higher purity, and the purity does not decrease; at the same time, the recovery rate is higher, and the sample loss is less. In addition, compared with the freeze-dried sample, the specific surface area of the air flow pulverized sample is larger, which is beneficial to faster dissolution and dissolution rate.

[0027] The pulverization process of the present application can realize step-by-step scaling from small-scale test, pilot-scale test to production, and can simultaneously consider product quality, yield, recovery rate, and efficiency. Attached Figure Description

[0028] Figure 1 shows typical particle size and particle size distribution spectra of the air-jet pulverized sample (top) of experimental group 14 and the freeze-dried sample (bottom) of experimental group 26.

[0029] Figure 2 shows a comparison of the properties of the air-jet pulverized sample (left) of experimental group 14, smegglutinin, and the freeze-dried sample (right) of experimental group 26.

[0030] Figure 3 shows a comparison of the purity test results of the air-jet pulverized sample (top) of experimental group 14 and the freeze-dried sample (bottom) of experimental group 26.

[0031] Figure 4 shows a comparison of the inherent dissolution curves of the air-jet pulverized sample (Group 14, Smegglutinin) and the freeze-dried sample (Group 26, Experimental Group 26). Detailed Implementation

[0032] The present invention will be further described below with reference to embodiments. It should be understood that the following embodiments are only used to explain the present invention and are not intended to limit the scope of protection of the present invention.

[0033] Example 1: Pulverization of Smegglutinin

[0034] S1) Freeze-drying: Obtaining smegglutinin freeze-dried powder by freeze-drying;

[0035] S2) Sieving: Take the lyophilized powder of smegglutinin and sieve it through a 10-200 mesh sieve to obtain the sieved powder of smegglutinin.

[0036] S3) Air jet milling: The sieved powder of smegglutide is processed by an air jet mill. The air source used in the air jet mill is compressed air, nitrogen or common inert gas. The pulverizing air pressure is set to 0.2 to 10 bar, and the pulverizing is carried out at a feed rate of 0.1 to 500 g / min. The final product is smegglutide powder with uniform properties, small particle size and narrow particle size distribution, which can meet the requirements of oral polypeptide solid dosage forms.

[0037] According to the general method described above, air jet milling experiments were conducted on smegglutinin. The specific sieve mesh size, milling air pressure, feed rate, and the results of yield, purity, maximum single impurity, particle size, and particle size distribution are shown in Table 1. The smegglutinin produced by air jet milling was a uniform powder with small particle size and narrow particle size distribution, a yield ≥95.0%, purity ≥99.0%, and single impurity <0.10%.

[0038] Table 1. Results of smegglutinin air jet milling experiments under different conditions

[0039] The comparison is as follows:

[0040] Comparative Example 1: Smeltide under different pulverization gas pressure

[0041] The pulverization gas pressure has an impact on the effect of air flow pulverization, and the results are shown in Table 2. When the pulverization gas pressure is lower than 0.2 bar, the pulverization effect is not obvious, and the particle size and particle size distribution are large; when it is higher than 10.0 bar, the yield is lower than 95%.

[0042] Table 2: Impact of different pulverization gas pressures on air flow pulverization of smeltide

[0043] Comparative Example 2: Impact of sieving treatment on air flow pulverization

[0044] The sample properties before air flow pulverization after lyophilization are not uniform, and the non-uniform sample properties have an impact on the effect of air flow pulverization, and the results are shown in Table 3. Although sieving treatment and direct air flow pulverization can obtain a sample with uniform properties, the particle size and particle size distribution are slightly larger. With the increase of the mesh number of the screen, the particle size and particle size distribution of the air flow pulverized sample gradually become smaller, but too large mesh number of the screen will lead to difficulty in sieving, time extension, and impact on efficiency.

[0045] Table 3: Impact of different mesh numbers of screens on air flow pulverization of smeltide

[0046] Comparative Example 3: Lyophilized sample of smeltide

[0047] The results are shown in Table 4. Compared with the air flow pulverized sample, the particle size and particle size distribution of the lyophilized sample are obviously larger, and the sample properties are not uniform.

[0048] Table 4: Test results of smeltide lyophilized powder

[0049] The specific surface area of smeltide prepared by different methods was analyzed, and the results are shown in Table 5. Compared with the lyophilized sample, the specific surface area of the air flow pulverized sample is larger, which is beneficial to faster dissolution and dissolution speed.

[0050] Table 5: Comparison results of specific surface area of smeltide air flow pulverized sample and lyophilized sample

[0051] The inherent dissolution of the sample was detected according to the pharmacopoeia guide (USP <1087>), and the results are shown in Table 6. Compared with the lyophilized sample, the dissolution amount of the air flow pulverized sample is more in a given time, and the inherent dissolution rate is faster.

[0052] Table 6: Inherent dissolution test results of smeltide air flow pulverized sample and lyophilized sample

[0053] The present invention relates to a Chinese meaning table for abbreviations

Claims

1. A method for pulverizing a polypeptide drug, characterized by, The pulverization method comprises the following steps: S1) freeze-drying: obtaining a polypeptide drug freeze-dried powder by freeze-drying; S2) sieving: taking the polypeptide drug freeze-dried powder, sieving the polypeptide drug freeze-dried powder with a 5-200 mesh sieve to obtain a sieved polypeptide drug sieved powder; S3) air flow pulverization: treating the polypeptide drug sieved powder with an air flow pulverizer to obtain a polypeptide drug powder; In the pulverization step, the pulverization air pressure is set to 1-10 bar.

2. The pulverization method according to claim 1, characterized by, The air source used in the air flow pulverizer is compressed air or inert gas.

3. The pulverization method according to claim 1, characterized by, The pulverization air pressure is set to 2-5 bar; Preferably, the pulverization air pressure is set to 2.5-3.5 bar; Preferably, the pulverization air pressure is set to 3 bar.

4. The pulverization method according to claim 1, characterized by, The sieve is 5 mesh, 10 mesh, 20 mesh, 50 mesh, 100 mesh, or 200 mesh.

5. The pulverization method according to claim 1, characterized by, The polypeptide drug is a GLP-1 analogue; Preferably, the GLP-1 analogue is selected from liraglutide, losemannide, dulaglutide, semeglutide, exenatide, albiglutide, medinadide, amycretin, rithaleride, CJC-1134-PC, BPI-3016, tirzepatide, exenatide, albiglutide, medinadide, amycretin, rithaleride.

6. The pulverization method according to claim 1, characterized by, The yield of the pulverization method is ≥95.0%. The yield is calculated as follows:

7. The pulverization method according to claim 1, characterized by, Yield / %=(obtained polypeptide drug powder / polypeptide drug freeze-dried powder)×100%. The particle size distribution of the polypeptide drug is D10 0.5-6 μm, D50 2-22 μm, and D90 4-35 μm.

8. The polypeptide drug powder obtained by the pulverization method according to any one of claims 1 to 7, characterized in that, Preferably, the particle size distribution of the polypeptide drug is D10 0.5-6 μm, D50 2-22 μm, and D90 4-35 μm. The polypeptide drug powder has a purity ≥99.5% and single impurities <0.10%.

9. The polypeptide drug powder of claim 8, characterized in that, Preferably, the particle size distribution of the polypeptide drug is D10 0.5-6 μm, D50 2-22 μm, and D90 4-35 μm. The particle size distribution of the polypeptide drug is D10 1-3 μm, D50 8-15 μm, and D90 15-28 μm.

10. Use of the polypeptide drug powder of claim 8 or 9 as a raw material drug in the preparation of an oral polypeptide solid drug.

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