Pharmaceutical composition containing recombinant human interleukin-2, pharmaceutical preparation and preparation method thereof

Abstract

The invention relates to the technical field of biological medicines, in particular to a pharmaceutical composition containing recombinant human interleukin-2, a pharmaceutical preparation and a preparation method of the pharmaceutical composition. Specifically, the pharmaceutical composition containing the recombinant human interleukin-2 is prepared from the following components in parts by weight: 0.05 to 0.3 part of recombinant human interleukin-2, 0.02 to 0.2 part of dissolution promoter, 0.1 to 0.4 part of sodium acetate, 0.2 to 0.8 part of acetic acid, 5 to 10 parts of sodium chloride and 500 to 2000 parts of water for injection, and the pH value of the recombinant human interleukin-2 solution is 3.6 to 4.5. By adopting the technical scheme, the stability of the recombinant human interleukin-2 solution can be improved, particularly, the effect is better under the scene of repeated use, and the product utilization rate is improved.

Description

Technical Field

[0001] The present application relates to the technical field of biopharmaceuticals, and in particular, to a pharmaceutical composition containing recombinant human interleukin-2, a pharmaceutical preparation and a preparation method thereof. Background Art

[0002] Interleukin-2 is a lymphokine produced by leukocytes. It is a multifunctional cytokine with anti-tumor, anti-viral and immunomodulatory effects. It is clinically used in the treatment of advanced renal cancer, malignant melanoma and cancerous pleural and abdominal effusions, and can also be tried in the comprehensive treatment of other malignant tumors. Low-dose interleukin-2 also has a good effect on immunotherapy of tuberous leprosy, mycoplasma infection, herpes and AIDS. It is one of the genetically engineered drugs with significant clinical efficacy.

[0003] Interleukin-2 is easily degraded in the presence of water and oxygen. According to previous reports, interleukin-2 can undergo chemical degradation and physical instability in solution. Although interleukin-2 injections have appeared in the prior art, the existing injections are difficult to preserve after one use, inconvenient to use and easy to cause waste.

[0004] In view of this, this application is filed. Summary of the invention

[0005] The purpose of the present application is to provide a pharmaceutical composition containing recombinant human interleukin-2, a pharmaceutical preparation and a preparation method thereof, so as to solve at least one technical problem described in the background technology.

[0006] Specifically, the first aspect of the present application provides a pharmaceutical composition containing recombinant human interleukin-2, comprising:

[0007] Recombinant human interleukin-20.05-0.3 copies,

[0008] 0.02-0.2 parts of solvent,

[0009] Sodium acetate 0.1-0.4 parts,

[0010] 0.2-0.8 parts of acetic acid,

[0011] Sodium chloride 5-10 parts,

[0012] Water for injection 500-2000 parts,

[0013] Wherein, the pH value of the recombinant human interleukin-2 solution is 3.6-4.5.

[0014] The above technical solution can improve the stability of the recombinant human interleukin-2 solution, especially its effect is better in the scenario of repeated use, thereby improving the utilization rate of the product.

[0015] Preferably, the pharmaceutical composition containing recombinant human interleukin-2 comprises the following components in parts by weight:

[0016] Recombinant human interleukin-20.1 copies,

[0017] 0.05 parts of solvent,

[0018] 0.15 parts of sodium acetate,

[0019] 0.5 parts of acetic acid,

[0020] 8 parts of sodium chloride,

[0021] 1000 parts of water for injection.

[0022] Preferably, the solubilizing agent is selected from at least one of polysorbate 80 and hydroxypropyl-β-cyclodextrin, wherein the degree of substitution of the hydroxypropyl-β-cyclodextrin is 0.6-10.0.

[0023] Preferably, the potency of the pharmaceutical composition containing recombinant human interleukin-2 is no higher than 1.5×10 6 IU / ml.

[0024] Preferably, the pharmaceutical composition containing recombinant human interleukin-2 further comprises the following components:

[0025] Mannitol 0.01-0.04 parts,

[0026] Methionine 400 0.01-0.02 parts.

[0027] Preferably, the pharmaceutical composition containing recombinant human interleukin-2 does not contain monosaccharides and / or amino acids. More preferably, the pharmaceutical composition containing recombinant human interleukin-2 does not contain glucose, fructose, arginine, or methionine.

[0028] Preferably, the pharmaceutical composition containing recombinant human interleukin-2 has a surface tension of 60-70 mN / m.

[0029] In a second aspect of the present application, a pharmaceutical preparation containing recombinant human interleukin-2 is provided, comprising:

[0030] The pharmaceutical composition of recombinant human interleukin-2 as described in the first aspect of the present application,

[0031] A container for directly storing a recombinant human interleukin-2 pharmaceutical composition,

[0032] The actuator is capable of performing a discharge operation on the recombinant human interleukin-2 pharmaceutical composition in the container according to the first discharge amount,

[0033] After a predetermined time has passed, the actuator can perform a discharging operation at a second discharging amount on the same container.

[0034] By adopting the above technical solution, the recombinant human interleukin-2 solution in the container can be discharged as needed for injection and other operations, and the remaining recombinant human interleukin-2 solution in the container can also be reused, thereby ensuring the ease of product operation and improving the utilization rate of the product.

[0035] Preferably, the container has a containing cavity made of a first material, and the containing cavity has a first end and a second end in its length direction;

[0036] The first end is provided with a cap body, the cap body having a membrane structure made of a second material, the membrane structure being capable of being pierced by a metal needle and restoring the closure of the first end after the metal needle is removed;

[0037] The second end is movably provided with a piston;

[0038] The cap body and the piston seal the interior of the container to form a sealing structure.

[0039] Preferably, the actuator comprises:

[0040] A receiving chamber, cooperating with the container, capable of fixing the container in the receiving chamber and exposing at least a portion of the cap of the container;

[0041] The push rod mechanism has a third end and a fourth end in its length direction, a pressing portion is provided at the third end, and a pressing plate is provided at the fourth end, and the push rod mechanism also includes a push rod connected to the pressing plate, when the pressing portion is pressed, the push rod is driven to drive the pressing plate to move along the length direction at the fourth end,

[0042] The pressure plate pushes the piston to move along the inner wall of the accommodating cavity to apply pressure to the recombinant human interleukin-2 pharmaceutical composition in the container.

[0043] Preferably, the actuator has an adjusting mechanism for controlling the stroke of the pressing portion to achieve the setting of the first discharge volume and the second discharge volume.

[0044] Furthermore, the actuator also includes a puncture needle, which can penetrate the membrane structure of the container and guide the recombinant human interleukin-2 pharmaceutical composition to be discharged when the recombinant human interleukin-2 solution is subjected to the pressure of the piston.

[0045] Preferably, the first material is medium borosilicate glass, and the silicone oil content is not higher than 0.2 mg / container. More preferably, the silicone oil content is 0.1-0.2 mg / container. The mass of the container is 15±2 g / container.

[0046] By adopting the above technical solution, when the concentration of the recombinant human interleukin-2 solution is controlled within a certain range and the container is made of a specific material, the recombinant human interleukin-2 has better stability in the container and can maintain good stability after multiple uses.

[0047] Preferably, the second material is nitrile rubber or chlorobutyl rubber.

[0048] Preferably, the pharmaceutical preparation further comprises:

[0049] Nitrogen, and the pressure in the container is 0.2-1.0Mpa.

[0050] The third aspect of the present application provides a method for preparing a pharmaceutical composition of recombinant human interleukin-2 as described in the first aspect of the present application, comprising the steps of:

[0051] Prepare a buffer solution, weigh sodium acetate according to the components described in the first aspect, add acetic acid and sodium chloride, and then add water for injection to prepare an acetate buffer solution;

[0052] Add a solvent in a corresponding proportion, wait for it to completely dissolve, and then filter to remove impurities in the solution;

[0053] Add recombinant human interleukin-2, add the solution obtained in the previous step to the recombinant human interleukin-2 lyophilized stock solution in proportion, filter after the protein is completely dissolved, and obtain a recombinant human interleukin-2 pharmaceutical composition.

[0054] Preferably, in the step of adding a dissolution agent, filtration is performed using a 0.22 μm filter membrane.

[0055] Preferably, in the step of adding recombinant human interleukin-2, filtration is performed using a 0.22 μm filter membrane.

[0056] Preferably, the time interval between the step of adding recombinant human interleukin-2 and the step of adding a dissolving agent does not exceed a second predetermined value, and the second predetermined value does not exceed 30 minutes.

[0057] Preferably, when the recombinant human interleukin-2 solution includes mannitol and methionine components, in the step of adding recombinant human interleukin-2, mannitol and methionine components are added first, and after the two are fully dissolved, the recombinant human interleukin-2 lyophilized stock solution is added.

[0058] In summary, this application has the following beneficial effects:

[0059] 1. The pharmaceutical composition containing recombinant human interleukin-2 provided in the present application can improve the stability of the recombinant human interleukin-2 solution, especially its effect is better in the scenario of repeated use, thereby achieving an increase in product utilization.

[0060] 2. The pharmaceutical preparation containing recombinant human interleukin-2 provided in the present application can improve the stability of recombinant human interleukin-2 in the container by controlling the concentration of the recombinant human interleukin-2 solution within a certain range and selecting a container made of a specific material, and can maintain good stability after multiple uses.

[0061] 3. The preparation method of the recombinant human interleukin-2 solution provided in the present application can improve the stability of the recombinant human interleukin-2 in the solution, and is more conducive to the multiple use of the pharmaceutical preparation. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 This is the activity change trend diagram of Scheme AD in the experimental example of this application under the stability test;

[0063] Figure 2 It is the activity change trend diagram of scheme AD in the experimental example of this application under accelerated test;

[0064] Figure 3 This is the activity change trend diagram of Scheme DG in the experimental example of this application under the stability test;

[0065] Figure 4 This is the activity change trend diagram of Scheme DG in the experimental example of this application under the accelerated test;

[0066] Figure 5 This is the activity change trend diagram of the scheme AFHIJ in the experimental example of this application under the stability test;

[0067] Figure 6 This is the activity change trend diagram of the scheme AFHIJ in the experimental example of this application under the accelerated test;

[0068] Figure 7 is a first angle schematic diagram of a container in an embodiment of the present application;

[0069] Figure 8 is a second angle schematic diagram of a container in one embodiment of the present application;

[0070] Fig. 9 This is a schematic diagram of the coordination state of a container and an actuator in one embodiment of the present application;

[0071] Fig.10is a schematic diagram of an actuator push rod mechanism in a first position in an embodiment of the present application;

[0072] Fig.11 is a schematic diagram of an actuator push rod mechanism in a second position in an embodiment of the present application;

[0073] Fig.12 is a schematic diagram of a puncture needle in one embodiment of the present application;

[0074] Fig.13 is a schematic diagram of the matching relationship between the puncture needle and the cap body in another embodiment of the present application;

[0075] Fig.14 It is a schematic diagram of a puncture needle in another embodiment of the present application.

[0076] Reference numerals:

[0077] 1. Container; 11. Cap body; 111. Membrane structure; 112. Second annular rib; 113. Second mark; 12. Piston; 21. Accommodating chamber; 22. Push rod mechanism; 221. Pressing portion; 23. Rod body; 24. Closing cap; 3. Puncture needle; 31. Second needle head; 32. First needle head; 33. Matching cavity; 331. First annular rib; 332. First mark. DETAILED DESCRIPTION

[0078] Exemplary embodiments will be described in detail herein, examples of which are shown in the accompanying drawings. When the following description refers to the drawings, the same numbers in different drawings represent the same or similar elements unless otherwise indicated. The implementations described in the following exemplary embodiments do not represent all implementations consistent with the present application. Instead, they are merely examples of devices and methods consistent with some aspects of the present application as detailed in the appended claims.

[0079] The terms used in this application are for the purpose of describing specific embodiments only and are not intended to limit this application. The singular forms of "a", "said" and "the" used in this application and the appended claims are also intended to include plural forms unless the context clearly indicates other meanings. It should also be understood that the term "and / or" used herein refers to and includes any or all possible combinations of one or more associated listed items.

[0080] The present application will be described in detail below through examples.

[0081] In the prior art, interleukin-2 injection is often used in anti-tumor, anti-viral or immunomodulatory treatments, but the existing injection is difficult to preserve after one use, is inconvenient to use and easily causes waste.

[0082] In view of this, the inventive concept of the present application is to provide a pharmaceutical preparation containing recombinant human interleukin-2 and a preparation method thereof, wherein the pharmaceutical preparation containing recombinant human interleukin-2 includes: a recombinant human interleukin-2 solution; a container for directly storing the recombinant human interleukin-2 solution; an actuator capable of setting a first discharge volume to perform a discharge operation on the recombinant human interleukin-2 solution in the container according to the first discharge volume; after a predetermined time, the actuator can perform a discharge operation on the same container according to the second discharge volume. By adopting the technical solution of the present application, the recombinant human interleukin-2 solution in the container can be discharged as needed for injection and other operations, and the remaining recombinant human interleukin-2 solution in the container can also be reused, which ensures that the product is easy to operate and improves the utilization rate of the product.

[0083] In order to better understand the above technical solution, the above technical solution will be described in detail below in conjunction with specific implementation methods. Those skilled in the art should also understand that the reaction time and drug feeding involved in this application cannot be absolutely accurate in the actual production process, but are all within the allowable error range. For example, if you expect to heat the sample for 30 minutes, the actual operation may be 1 second more or less than 1 second in 30 minutes; if you expect to weigh 30g of sample, you may actually weigh 30.001g or 29.998g.

[0084] Experimental Example 1 Stability Test

[0085] The container containing the recombinant human interleukin-2 solution was stored in the dark for a long time at 25±2℃ and relative humidity of 60±5%RH. Every 7 days, the rubber stopper was penetrated by an injection needle without contacting the recombinant human interleukin-2 solution for 1 minute, and the light of Lx was 1000±100Lx to simulate the multiple application scenario of this product. The activity of interleukin-2 was measured at 0th month, 3rd month, 6th month, 9th month, and 12th month.

[0086] Among them, the container has a maximum capacity of 3ml and a weight of 15±2g.

[0087] Experimental Example 2 Accelerated Test

[0088] The container containing the recombinant human interleukin-2 solution was stored in the dark for a long time at 37±2℃ and relative humidity 60±5%RH. Every 7 days, the rubber stopper was penetrated by an injection needle without contacting the recombinant human interleukin-2 solution for 1 minute, and the light of Lx was 1000±100Lx to simulate the multiple application scenario of this product. The activity of interleukin-2 was measured at 0th month, 1st month, 3rd month, and 6th month.

[0089] Among them, the container has a maximum capacity of 3ml and a weight of 15±2g.

[0090] Experimental Example 3: Lighting Test

[0091] The container containing the recombinant human interleukin-2 solution was stored in the dark for a long time at 4±2°C, 60±5%RH, and 1000±100Lx of light.

[0092] Experimental Example 4

[0093] This experimental example tests the interleukin-2 activity of various experimental schemes under stability test and accelerated test.

[0094] Plan A

[0095] In the step of preparing a buffer solution, 0.15 parts of sodium acetate are weighed, 0.5 parts of acetic acid and 8 parts of sodium chloride are added, and then 1000 parts of water for injection are added to prepare an acetate buffer solution;

[0096] In the step of adding a dissolution promoter, 0.05 parts of polysorbate 80 are added, and after it is completely dissolved, it is filtered to remove impurities in the solution;

[0097] In the step of adding recombinant human interleukin-2, 0.1 parts of recombinant human interleukin-2 lyophilized stock solution are added to the solution obtained in the previous step, and the solution is filtered after the protein is completely dissolved to obtain a recombinant human interleukin-2 solution.

[0098] 0.8 ml of the recombinant human interleukin-2 solution obtained above was injected into a container made of medium borosilicate glass, and the silicone oil content of the container was 0.6 mg / container.

[0099] Plan B

[0100] In the step of preparing a buffer solution, 0.15 parts of sodium acetate are weighed, 0.5 parts of acetic acid and 8 parts of sodium chloride are added, and then 1000 parts of water for injection are added to prepare an acetate buffer solution;

[0101] In the step of adding a dissolution promoter, 0.05 parts of polysorbate 80 are added, and after it is completely dissolved, it is filtered to remove impurities in the solution;

[0102] In the step of adding recombinant human interleukin-2, 0.02 parts of mannitol are added to the solution obtained in the previous step, and after it is fully dissolved, 0.1 parts of recombinant human interleukin-2 lyophilized stock solution are added, and after the protein is completely dissolved, the solution is filtered to obtain a recombinant human interleukin-2 solution.

[0103] 0.8 ml of the recombinant human interleukin-2 solution obtained above was injected into a container made of medium borosilicate glass, and the silicone oil content of the container was 0.6 mg / container.

[0104] Plan C

[0105] In the step of preparing a buffer solution, 0.15 parts of sodium acetate are weighed, 0.5 parts of acetic acid and 8 parts of sodium chloride are added, and then 1000 parts of water for injection are added to prepare an acetate buffer solution;

[0106] In the step of adding a dissolution promoter, 0.05 parts of polysorbate 80 are added, and after it is completely dissolved, it is filtered to remove impurities in the solution;

[0107] In the step of adding recombinant human interleukin-2, 0.01 parts of methionine are added to the solution obtained in the previous step, and after it is fully dissolved, 0.1 parts of recombinant human interleukin-2 lyophilized stock solution are added. After the protein is completely dissolved, it is filtered to obtain a recombinant human interleukin-2 solution.

[0108] 0.8 ml of the recombinant human interleukin-2 solution obtained above was injected into a container made of medium borosilicate glass, and the silicone oil content of the container was 0.6 mg / container.

[0109] Plan D

[0110] In the step of preparing a buffer solution, 0.15 parts of sodium acetate are weighed, 0.5 parts of acetic acid and 8 parts of sodium chloride are added, and then 1000 parts of water for injection are added to prepare an acetate buffer solution;

[0111] In the step of adding a dissolution promoter, 0.05 parts of polysorbate 80 are added, and after it is completely dissolved, it is filtered to remove impurities in the solution;

[0112] In the step of adding recombinant human interleukin-2, 0.02 parts of mannitol and 0.01 parts of methionine are added to the solution obtained in the previous step in sequence, and after they are fully dissolved, 0.1 parts of recombinant human interleukin-2 lyophilized stock solution is added, and after the protein is completely dissolved, the solution is filtered to obtain a recombinant human interleukin-2 solution.

[0113] 0.8 ml of the recombinant human interleukin-2 solution obtained above was injected into a container made of medium borosilicate glass, and the silicone oil content of the container was 0.6 mg / container.

[0114] Schemes A to D were tested for interleukin-2 activity after stability test and accelerated test according to the methods of Experimental Example 1 and Experimental Example 2, respectively. The test data are as follows: Figure 1 , Figure 2 shown.

[0115] Depend on Figure 1 , Figure 2 The experimental results show that:

[0116] First, the activity of Scheme AD decreased rapidly in both stability and accelerated tests, indicating that the stability of the product is easily adversely affected in a multiple-use test environment;

[0117] Second, the activity of Scheme D decreased more slowly than that of Schemes AC in both stability tests and accelerated tests, and its stability was better. This indicates that adding appropriate amounts of mannitol and methionine components to the recombinant human interleukin-2 solution can improve the stability of the product in multiple reuse scenarios to a certain extent.

[0118] Experimental Example 5

[0119] This experimental example tests the interleukin-2 activity of various experimental schemes under stability test and accelerated test.

[0120] Plan E

[0121] Plan E is basically the same as Plan D, except that:

[0122] 0.8 ml of the obtained recombinant human interleukin-2 solution was injected into a container made of medium borosilicate glass, and the silicone oil content of the container was 0.3 mg / container.

[0123] Plan F

[0124] Plan F is basically the same as Plan D, except that:

[0125] 0.8 ml of the obtained recombinant human interleukin-2 solution was injected into a container made of medium borosilicate glass with a silicone oil content of 0.1 mg / container.

[0126] Plan G

[0127] Plan G is basically the same as Plan D, except that:

[0128] 0.8 ml of the obtained recombinant human interleukin-2 solution was injected into a container made of medium borosilicate glass, and the silicone oil content of the container was 0.05 mg / container.

[0129] Scheme H

[0130] Plan H is basically the same as Plan A, except that:

[0131] 0.8 ml of the obtained recombinant human interleukin-2 solution was injected into a container made of medium borosilicate glass with a silicone oil content of 0.1 mg / container.

[0132] Scheme I

[0133] Plan I is basically the same as Plan A, except that:

[0134] In the step of adding recombinant human interleukin-2, 0.2 parts of recombinant human interleukin-2 lyophilized stock solution was added.

[0135] Plan J

[0136] Plan J is basically the same as Plan A, except that:

[0137] In the step of adding recombinant human interleukin-2, 0.2 parts of recombinant human interleukin-2 lyophilized stock solution were added;

[0138] The obtained recombinant human interleukin-2 solution was injected into a 0.8 ml container made of medium borosilicate glass, and the silicone oil content of the container was 0.1 mg / container.

[0139] Scheme D to Scheme G were tested for interleukin-2 activity after stability test and accelerated test according to the methods of Experimental Example 1 and Experimental Example 2, respectively. The test data are as follows Figure 3 , Figure 4 As shown;

[0140] Scheme A, Scheme F, Scheme H-Scheme I were tested for interleukin-2 activity after stability test and accelerated test according to the methods of Experimental Example 1 and Experimental Example 2. The test data are as follows Figure 5 , Figure 6 shown.

[0141] Depend on Figure 3-Figure 6 The experimental results show that:

[0142] First, reference Figure 3-Figure 4 Schemes D and G are mainly based on the different silicone oil contents of the container glass. It can be found that when the silicone oil content is low, the stability of interleukin-2 will be improved (such as Schemes F and G in the stability experiment); but when the silicone oil content is less than 0.1 mg, the stability does not show a significant improvement. This may be because the silicone oil content is too low, which leads to a decrease in the sliding properties of the product during use, thereby reducing the airtightness of the container and the rubber stopper.

[0143] Second, reference Figure 5-Figure 6 The results of stability tests and accelerated tests of Scheme A and Scheme H showed that under the formulation of Scheme A, the reduction in silicone oil content had little effect on the stability of interleukin-2; indicating that the reduction in silicone oil content under the formulation of Scheme D had a better effect on improving the stability of interleukin-2.

[0144] Third, reference Figure 5-Figure 6 The results of the stability test and accelerated test of Scheme I and Scheme J showed that when the interleukin-2 content was high, even if the components did not contain mannitol and methionine, the reduction in the silicone oil content would have a positive effect on the stability of interleukin-2. However, at this time, the activity of interleukin-2 decreased to a high degree, resulting in product waste and low utilization efficiency.

[0145] Therefore, based on the above experimental results, the potency of the recombinant human interleukin-2 solution in the product is controlled to be no higher than 1.5×10 6IU / ml, and the silicone oil content is controlled at 0.1-0.2 mg / container, at this time, a recombinant human interleukin-2 preparation with better stability can be obtained.

[0146] Example 1

[0147] This embodiment provides a container and an actuator that can be used in the present application.

[0148] Specifically, refer to Figure 7-Figure 8 The container 1 includes a first end provided with a cap body 11, and a second end provided with a piston 12. The piston 12 can slide along the length direction of the container 1 at the second end and seal the second end. A membrane structure 111 is provided inside the cap body 11 and seals the first end. The membrane structure can be made of nitrile rubber or chlorobutyl rubber. The membrane structure can be punctured and restore the sealing of the first end after the puncture device is removed. Therefore, the cap body 11 and the piston 12 seal the inside of the container 1 to form a sealing structure for storing the recombinant human interleukin-2 solution.

[0149] Further, refer to Figure 9-11 The actuator includes a storage chamber 21, a rod body 23, a push rod mechanism 22, a closure cap 24, and has an output end and a matching end in its length direction. The storage chamber 21 can be used to place the container 1, and after placing the container 1, the cap body 11 is partially exposed outside the output end to match the puncture needle. The storage chamber 21 is connected to the rod body 23 at its matching end, and the rod body 23 has a cavity for accommodating the push rod mechanism 22. The push rod mechanism 22 has a third end and a fourth end in its length, which is movably arranged at one end of the rod body 23 and penetrates the cavity of the rod body 23 to push the piston 12; further, a pressing portion 221 is arranged at the third end, and a pressing plate is arranged at the fourth end. The push rod mechanism also includes a push rod connected to the pressing plate. When the pressing portion 221 is pressed, the push rod is driven to drive the pressing plate to move along the length direction at the fourth end, and the pressing plate pushes the piston 12 to move along the inner wall of the cavity of the container 1 to apply pressure to the recombinant human interleukin-2 solution in the container 1. The closure cap 24 is used to be sleeved on the outside of the containing chamber 21 to play a role of protection and light shielding.

[0150] Continue to refer Figure 10-12The actuator also includes a puncture needle 3, which has a matching cavity 33, which matches with the cap body 11. In some embodiments, the matching cavity 33 is provided with a thread inside, which matches with the thread outside the cap body 11; a first needle 32 is extended from the matching cavity 33, and the first needle 32 can puncture the membrane structure 11 of the cap body when the matching cavity 33 matches with the cap body 11; a second needle 31 is provided at the end of the puncture needle 3 away from the first needle 32, and the second needle 31 is used to puncture the human body and inject the recombinant human interleukin-2 solution.

[0151] In some embodiments, Figure 10-11 As shown, since the recombinant human interleukin-2 solution in the container 1 of the present application may be used multiple times, the same position of the membrane structure 111 inside the cap body 1 may be deformed when punctured multiple times, resulting in the inability to seal the container very tightly.

[0152] Based on this, the present application proposes a Fig.13 The puncture needle 3, its first needle head 32 forms an angle with the horizontal position, and the angle is usually 5-10°. In this way, when the membrane structure 111 is repeatedly punctured, the puncture needle 3 can be rotated to different angles to prevent multiple punctures of the same position of the membrane structure 111, thereby ensuring the sealing of the product. In this embodiment, the puncture needle and the cap body 11 are matched in a snap-on manner. A first annular convex rib 331 is provided inside the puncture needle, and a second annular convex rib 112 that matches the first annular convex rib 331 is also provided on the outer wall of the cap body 11, thereby achieving snap-on connection. In order to further avoid multiple punctures of the same position of the membrane structure 111, the puncture needle and the outer wall of the cap body are respectively provided with a first mark 332 and a second mark 113. The user records the marks of each alignment, which can further avoid puncturing the same position of the membrane structure 111. In this embodiment, the first needle head 32 can also be other alternative forms, such as Fig.14 The bent structure shown.

[0153] In the present application, when operating the actuator, the recombinant human interleukin-2 solution in the container 1 can be discharged according to the first discharge volume; after the operation, the container 1 with the remaining recombinant human interleukin-2 solution can be preserved; after a period of time, if a discharge operation such as injection is still required, the container 1 can be reinstalled, and the actuator can be used to perform a discharge operation of the second discharge volume on the same container 1.

[0154] Example 2

[0155] In the step of preparing a buffer solution, 0.1 parts of sodium acetate are weighed, 0.2 parts of acetic acid and 5 parts of sodium chloride are added, and then 500 parts of water for injection are added to prepare an acetate buffer solution;

[0156] In the step of adding a dissolution promoter, 0.02 parts of polysorbate 80 are added, and after it is completely dissolved, it is filtered to remove impurities in the solution;

[0157] In the step of adding recombinant human interleukin-2, 0.05 parts of recombinant human interleukin-2 lyophilized stock solution are added to the solution obtained in the previous step, and the recombinant human interleukin-2 solution is obtained by filtering after the protein is completely dissolved.

[0158] The recombinant human interleukin-2 solution obtained above was injected into the container shown in Example 1, and the maximum capacity of the container was 3 ml, and 0.8 ml was injected into the container; the container was made of medium borosilicate glass, and the silicone oil content was 0.6 mg / container. The cap membrane structure of the container was made of chlorobutyl rubber.

[0159] Example 3

[0160] In the step of preparing a buffer solution, 0.15 parts of sodium acetate are weighed, 0.5 parts of acetic acid and 8 parts of sodium chloride are added, and then 1000 parts of water for injection are added to prepare an acetate buffer solution;

[0161] In the step of adding a dissolution promoter, 0.05 parts of polysorbate 80 are added, and after it is completely dissolved, it is filtered to remove impurities in the solution;

[0162] In the step of adding recombinant human interleukin-2, 0.1 parts of recombinant human interleukin-2 lyophilized stock solution are added to the solution obtained in the previous step, and the solution is filtered after the protein is completely dissolved to obtain a recombinant human interleukin-2 solution.

[0163] The recombinant human interleukin-2 solution obtained above was injected into the container shown in Example 1, and the maximum capacity of the container was 3 ml, and 0.8 ml was injected into the container; the container was made of medium borosilicate glass, and the silicone oil content was 0.6 mg / container. The cap membrane structure of the container was made of chlorobutyl rubber.

[0164] Example 4

[0165] In the step of preparing a buffer solution, 0.4 parts of sodium acetate are weighed, 0.8 parts of acetic acid and 10 parts of sodium chloride are added, and then 2000 parts of water for injection are added to prepare an acetate buffer solution;

[0166] In the step of adding a dissolution promoter, 0.2 parts of polysorbate 80 are added, and after it is completely dissolved, it is filtered to remove impurities in the solution;

[0167] In the step of adding recombinant human interleukin-2, 0.3 parts of recombinant human interleukin-2 lyophilized stock solution are added to the solution obtained in the previous step, and the solution is filtered after the protein is completely dissolved to obtain a recombinant human interleukin-2 solution.

[0168] The recombinant human interleukin-2 solution obtained above was injected into the container shown in Example 1, and the maximum capacity of the container was 3 ml, and 0.8 ml was injected into the container; the container was made of medium borosilicate glass, and the silicone oil content was 0.6 mg / container. The cap membrane structure of the container was made of chlorobutyl rubber.

[0169] Example 5

[0170] In the step of preparing a buffer solution, 0.1 parts of sodium acetate are weighed, 0.2 parts of acetic acid and 5 parts of sodium chloride are added, and then 500 parts of water for injection are added to prepare an acetate buffer solution;

[0171] In the step of adding a dissolution promoter, 0.02 parts of polysorbate 80 are added, and after it is completely dissolved, it is filtered to remove impurities in the solution;

[0172] In the step of adding recombinant human interleukin-2, 0.01 parts of mannitol and 0.01 parts of methionine are added to the solution obtained in the previous step in sequence, and after they are fully dissolved, 0.05 parts of recombinant human interleukin-2 lyophilized stock solution are added. After the protein is completely dissolved, the solution is filtered to obtain a recombinant human interleukin-2 solution.

[0173] The recombinant human interleukin-2 solution obtained above was injected into the container shown in Example 1, and the maximum capacity of the container was 3 ml, and 0.8 ml was injected into the container; the container was made of medium borosilicate glass, and the silicone oil content was 0.6 mg / container. The cap membrane structure of the container was made of chlorobutyl rubber.

[0174] Example 6

[0175] In the step of preparing a buffer solution, 0.15 parts of sodium acetate are weighed, 0.5 parts of acetic acid and 8 parts of sodium chloride are added, and then 1000 parts of water for injection are added to prepare an acetate buffer solution;

[0176] In the step of adding a dissolution promoter, 0.05 parts of polysorbate 80 are added, and after it is completely dissolved, it is filtered to remove impurities in the solution;

[0177] In the step of adding recombinant human interleukin-2, 0.02 parts of mannitol and 0.01 parts of methionine are added to the solution obtained in the previous step in sequence, and after they are fully dissolved, 0.1 parts of recombinant human interleukin-2 lyophilized stock solution is added, and after the protein is completely dissolved, the solution is filtered to obtain a recombinant human interleukin-2 solution.

[0178] The recombinant human interleukin-2 solution obtained above was injected into the container shown in Example 1, and the maximum capacity of the container was 3 ml, and 0.8 ml was injected into the container; the container was made of medium borosilicate glass, and the silicone oil content was 0.6 mg / container. The cap membrane structure of the container was made of chlorobutyl rubber.

[0179] Example 7

[0180] In the step of preparing a buffer solution, 0.4 parts of sodium acetate are weighed, 0.8 parts of acetic acid and 10 parts of sodium chloride are added, and then 2000 parts of water for injection are added to prepare an acetate buffer solution;

[0181] In the step of adding a dissolution promoter, 0.2 parts of polysorbate 80 were added, and after it was completely dissolved, it was filtered to remove impurities in the solution;

[0182] In the step of adding recombinant human interleukin-2, 0.04 parts of mannitol and 0.02 parts of methionine are added to the solution obtained in the previous step in sequence, and after they are fully dissolved, 0.3 parts of recombinant human interleukin-2 lyophilized stock solution are added. After the protein is completely dissolved, the solution is filtered to obtain a recombinant human interleukin-2 solution.

[0183] The recombinant human interleukin-2 solution obtained above was injected into the container shown in Example 1, and the maximum capacity of the container was 3 ml, and 0.8 ml was injected into the container; the container was made of medium borosilicate glass, and the silicone oil content was 0.6 mg / container. The cap membrane structure of the container was made of chlorobutyl rubber.

[0184] Example 8

[0185] The technical solution of Example 8 is basically the same as that of Example 6, except that:

[0186] The container is made of medium borosilicate glass and its silicone oil content is 0.2 mg / container.

[0187] Example 9

[0188] The technical solution of Example 10 is basically the same as that of Example 6, except that:

[0189] The container is made of medium borosilicate glass and its silicone oil content is 0.1 mg / container.

[0190] Example 10

[0191] The technical solution of Example 10 is basically the same as that of Example 9, except that:

[0192] Nitrogen was added into the container to make the pressure inside the container 0.2 MPa.

[0193] Embodiment 11

[0194] The technical solution of Example 11 is basically the same as that of Example 9, except that:

[0195] Nitrogen was added into the container to make the pressure inside the container 1.0 MPa.

[0196] Comparative Example 1

[0197] The technical solution of Comparative Example 1 is basically the same as that of Example 2, except that:

[0198] In the step of adding recombinant human interleukin-2, 0.5 parts of recombinant human interleukin-2 lyophilized stock solution was added.

[0199] Comparative Example 2

[0200] The technical solution of Comparative Example 2 is basically the same as that of Example 9, except that:

[0201] In the step of adding recombinant human interleukin-2, 0.02 parts of mannitol, 0.01 parts of methionine, and 0.02 parts of arginine are added to the solution obtained in the previous step in sequence, and after they are fully dissolved, 0.1 parts of recombinant human interleukin-2 lyophilized stock solution is added, and after the protein is completely dissolved, it is filtered to obtain a recombinant human interleukin-2 solution.

[0202] Comparative Example 3

[0203] The technical solution of Comparative Example 3 is basically the same as that of Example 9, except that:

[0204] In the step of adding recombinant human interleukin-2, 0.02 parts of mannitol, 0.01 parts of methionine, and 0.02 parts of methionine are added to the solution obtained in the previous step in sequence, and after it is fully dissolved, 0.1 parts of recombinant human interleukin-2 lyophilized stock solution is added, and after the protein is completely dissolved, it is filtered to obtain a recombinant human interleukin-2 solution.

[0205] Comparative Example 4

[0206] The technical solution of Comparative Example 4 is basically the same as that of Example 9, except that:

[0207] In the step of adding recombinant human interleukin-2, 0.02 parts of mannitol, 0.01 parts of methionine, and 0.02 parts of glucose are added to the solution obtained in the previous step in sequence, and after they are fully dissolved, 0.1 parts of recombinant human interleukin-2 lyophilized stock solution is added, and after the protein is completely dissolved, it is filtered to obtain a recombinant human interleukin-2 solution.

[0208] Comparative Example 5

[0209] The technical solution of Comparative Example 5 is basically the same as that of Example 9, except that:

[0210] In the step of adding recombinant human interleukin-2, 0.02 parts of mannitol, 0.01 parts of methionine, and 0.02 parts of fructose are added to the solution obtained in the previous step in sequence, and after they are fully dissolved, 0.1 parts of recombinant human interleukin-2 lyophilized stock solution is added, and after the protein is completely dissolved, it is filtered to obtain a recombinant human interleukin-2 solution.

[0211] Example 12

[0212] The properties and pH at day 0 in Examples 2-11 and Comparative Examples 1-5 were measured, and the experimental results are shown in Table 1.

[0213] The interleukin-2 biological activity of Examples 2-12 and Comparative Examples 1-5 after standing for 12 months was determined according to the method of Experimental Example 1; the interleukin-2 biological activity of Examples 2-12 and Comparative Examples 1-5 after standing for 12 months was determined according to the method of Experimental Example 3; the experimental results are shown in Table 2.

[0214] Table 1 Test results of Examples 2-11 and Comparative Examples 1-5

[0215]

[0216]

[0217] According to the results in Table 1, in the examples provided in this application, the properties and pH of the products meet the quality standards of the products and have relatively stable quality.

[0218] Table 2 Test results of Examples 2-11 and Comparative Examples 1-5

[0219]

[0220]

[0221] According to the experimental results in Table 2, for the stability test:

[0222] First, comparing Example 2 with Comparative Example 1, when the content of recombinant human interleukin-2 in the component is too high, the activity loss is high in the use scenario of the product of the present application, so the content of recombinant human interleukin-2 should be controlled within an appropriate range; comparing Example 4 with Examples 2-3, and Example 7 with Examples 5-6, although the activity loss rate of the product with a high recombinant human interleukin-2 concentration is also slightly higher, it is already controlled at a lower level compared to Comparative Example 1;

[0223] Second, by comparing Example 5-7 with Example 2-4, the activity loss rate of Example 5-7 is significantly lower than that of Example 2-4, indicating that the addition of mannitol and methionine components can reduce the activity loss rate of the product at an appropriate recombinant human interleukin-2 content;

[0224] Third, comparing Examples 8-11 with Examples 5-7, the activity loss rate of Examples 8-11 is lower than that of Examples 5-7, indicating that reducing the silicone oil content of the container can reduce the activity loss rate of the product;

[0225] Fourth, comparing Example 9 with Comparative Examples 2-5, the activity loss rate of Comparative Example 2-3 increased significantly, indicating that the addition of other components to the components may affect the stability of the product. Comparative Examples 2-3 are two amino acids, and Comparative Examples 4-5 are two monosaccharides, so the addition of amino acids and monosaccharides to the components may affect the stability of the product.

[0226] In addition, according to the experimental results in Table 2, for the illumination test:

[0227] First, the embodiments provided in this application are relatively stable under light exposure tests;

[0228] Second, comparing Example 7 with Example 4, when the components include mannitol and 0.01 parts of methionine, the high-concentration product is more sensitive to light and can be considered to be stored in a light-proof environment.

[0229] In summary, the pharmaceutical preparation containing recombinant human interleukin-2 provided in the present application can discharge the recombinant human interleukin-2 solution in the container as needed for injection and other operations, and the remaining recombinant human interleukin-2 solution in the container can also be reused, thereby ensuring that the product is easy to operate and improving the utilization rate of the product, and has a good commercial prospect.

[0230] For ordinary technicians in this field, the technical features in the above embodiments can be freely combined, and the formed technical solutions also belong to the embodiments disclosed in this application.

[0231] Furthermore, without departing from the principles of the present application, several improvements and modifications may be made to the present application, and these improvements and modifications also fall within the protection scope of the claims of the present application.

Claims

1. A pharmaceutical composition containing recombinant human interleukin-2, characterized in that: include: Recombinant human interleukin-2 0.05-0.3 copies, 0.02-0.2 parts of solvent, Sodium acetate 0.1-0.4 parts, 0.2-0.8 parts of acetic acid, Sodium chloride 5-10 parts, Water for injection 500-2000 parts, Wherein, the pH value of the recombinant human interleukin-2 solution is 3.6-4.

5.

2. The pharmaceutical composition according to claim 1, characterized in that: The pharmaceutical composition containing recombinant human interleukin-2 comprises the following components in parts by weight: Recombinant human interleukin-2 0.1 parts, 0.05 parts of solvent, 0.15 parts of sodium acetate, 0.5 parts of acetic acid, 8 parts of sodium chloride, 1000 parts of water for injection.

3. The pharmaceutical composition according to claim 1 or 2, characterized in that: The dissolution promoter is selected from at least one of polysorbate 80 and hydroxypropyl-β-cyclodextrin, wherein the degree of substitution of the hydroxypropyl-β-cyclodextrin is 0.6-10.

0.

4. The pharmaceutical composition according to claim 3, characterized in that: The pharmaceutical composition containing recombinant human interleukin-2 also includes the following components: Mannitol 0.01-0.04 parts, Methionine 4000.01-0.02 parts.

5. The pharmaceutical composition according to claim 3, characterized in that: The pharmaceutical composition containing recombinant human interleukin-2 does not contain monosaccharides and / or amino acids.

6. A pharmaceutical preparation containing recombinant human interleukin-2, characterized in that: include: The pharmaceutical composition of recombinant human interleukin-2 according to any one of claims 1 to 5, A container for directly storing a recombinant human interleukin-2 pharmaceutical composition, The actuator is capable of performing a discharge operation on the recombinant human interleukin-2 pharmaceutical composition in the container according to the first discharge amount, After a predetermined time has passed, the actuator can perform a discharging operation at a second discharging amount on the same container.

7. The pharmaceutical preparation according to claim 6, characterized in that: The container has a containing cavity made of a first material, and the containing cavity has a first end and a second end in its length direction; The first end is provided with a cap body, the cap body having a membrane structure made of a second material, the membrane structure being capable of being pierced by a metal needle and restoring the closure of the first end after the metal needle is removed; The second end is movably provided with a piston; The cap body and the piston seal the interior of the container to form a sealing structure.

8. The pharmaceutical preparation according to claim 7, characterized in that: The actuator comprises: A receiving chamber, cooperating with the container, capable of fixing the container in the receiving chamber and exposing at least a portion of the cap of the container; The push rod mechanism has a third end and a fourth end in its length direction, a pressing portion is provided at the third end, and a pressing plate is provided at the fourth end, and the push rod mechanism also includes a push rod connected to the pressing plate, when the pressing portion is pressed, the push rod is driven to drive the pressing plate to move along the length direction at the fourth end, The pressure plate pushes the piston to move along the inner wall of the accommodating cavity to apply pressure to the recombinant human interleukin-2 pharmaceutical composition in the container.

9. The pharmaceutical preparation according to claim 8, characterized in that: The actuator has an adjusting mechanism for controlling the stroke of the pressing portion to achieve the setting of the first discharge volume and the second discharge volume.

10. A method for preparing the pharmaceutical composition of recombinant human interleukin-2 according to any one of claims 1 to 5, comprising the steps of: Prepare a buffer solution, weigh sodium acetate according to the components described in any one of claims 1 to 5, add acetic acid and sodium chloride, and then add water for injection to prepare an acetate buffer solution; Add a solvent in a corresponding proportion, wait for it to completely dissolve, and then filter to remove impurities in the solution; Add recombinant human interleukin-2, add the solution obtained in the previous step to the recombinant human interleukin-2 lyophilized stock solution in proportion, filter after the protein is completely dissolved, and obtain a recombinant human interleukin-2 pharmaceutical composition.

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