Process for manufacturing orally disintegrating tablet comprising cytokine

Abstract

To provide a process for manufacturing an orally disintegrating tablet (ODT) comprising a cytokine as an active pharmaceutical ingredient.SOLUTION: The present invention provides a process for manufacturing an orally disintegrating tablet (ODT) comprising a cytokine as an active pharmaceutical ingredient, comprising: acidifying an excipient; conducting a first granulation step of the acidified excipient to obtain acidic powders; and conducting a second granulation step by mixing the acidic powders and the cytokine to obtain granules containing the cytokine. The present application also provides an ODT manufactured by the process.SELECTED DRAWING: Figure 3

Description

[Technical field]

[0001] The present application relates to a method for producing an orally disintegrating tablet (ODT), and in particular to a method for producing an ODT that includes a cytokine. [Background technology]

[0002] Cytokines are small molecular weight proteins (approximately 6 to 70 kDa) that are important in cell signaling. Due to their size, cytokines cannot cross the lipid bilayer of cells and enter the cytoplasm, and therefore typically exert their function by interacting with specific cytokine receptors on the surface of target cells. Cytokines have been shown to participate in autocrine, paracrine, and endocrine signaling as immunomodulatory agents.

[0003] Cytokines include chemokines, interferons, interleukins, lymphokines, and tumor necrosis factors. Cytokines are produced by a wide range of cells, including immune cells such as macrophages, B lymphocytes, T lymphocytes, and mast cells, as well as endothelial cells, fibroblasts, and various stromal cells. A given cytokine may be produced by more than one type of cell. They act through cell surface receptors and are particularly important in the immune system; cytokines regulate the balance between humoral and cellular immune responses, as well as control the maturation, proliferation, and responsiveness of specific cell populations. Some cytokines enhance or inhibit the action of other cytokines in complex ways. Cytokines are important in health and disease, particularly in the host immune response to infection, inflammation, trauma, sepsis, cancer, and reproduction.

[0004] Interferons (IFNs) are proteins made by host cells in response to the presence of pathogens such as viruses, bacteria, parasites, or tumor cells. IFNs enable the exchange of information between cells to trigger protective defenses of the immune system that eradicate the pathogen or tumor. Of the nine different human interferon families that have been identified, interferon-alpha (IFN-α) has been the most widely studied. The US FDA has approved the use of high doses of IFNα given by injection to treat hepatitis B and C as well as some cancers. High doses of interferon, for example more than 1 million IU, are traditionally administered by intramuscular injection to treat symptoms systemically.

[0005] In recent years, published studies have shown that oral administration of interferon activates dozens of immune system genes in peripheral blood without the side effects associated with high-dose injections, and is effective against viral and autoimmune diseases. However, the protein stability of orally administered interferon remains a challenge.

[0006] Oral administration can be via ODTs, etc., which can be rapidly disintegrated with saliva or a small amount of water. Compliance can be increased by the application of ODTs, as ODTs are convenient and easy to administer for patients. However, ODTs are a newly developed dosage form, and some technical issues, such as tablet properties, still exist.

[0007] As such, there remains a need for ODTs containing cytokines such as interferons that have good protein stability. Summary of the Invention

[0008] The present application provides a method for producing an ODT comprising a cytokine as an active ingredient, the method comprising: acidifying an excipient, carrying out a first granulation step of the acidified excipient to obtain an acidic powder, and carrying out a second granulation step by mixing the acidic powder with the cytokine to obtain granules containing the cytokine.

[0009] The present application also provides a method for preparing an ODT comprising a cytokine as an active ingredient, the method comprising: acidifying a first excipient; performing a first granulation step of the acidified first excipient to obtain a first acidic powder; sieving and granulating the first acidic powder to obtain a second acidic powder; and performing a second granulation step by mixing the second acidic powder with the cytokine to obtain a first granule. [Brief description of the drawings]

[0010] [Figure 1] FIG. 1 illustrates a method for producing an ODT containing a cytokine as an active ingredient according to one embodiment of the present application. [Diagram 2] FIG. 1 shows the protective effect of various agents against IFN-α. [Diagram 3] FIG. 1 shows the results of a study on the effect of the excipient acidification step of the method of the present application. IFN-α content was measured using ELISA at OD450, and five samples in each group were labeled as acidified (S1-S5) and not acidified (S1-S5). [Figure 4] FIG. 13 shows test results regarding the effect of a treatment step on a first acidic powder. [Diagram 5] FIG. 13 shows test results regarding the effectiveness of the device used in the second granulation step. [Figure 6] FIG. 1 shows the results of a study on the effect of tablet thickness on IFN-α potency. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] The present application provides a method for producing an ODT that contains a cytokine as an active ingredient.

[0012] The cytokine is selected from the group including chemokines, interferons, interleukins, lymphokines, and tumor necrosis factors. The cytokine can be a natural cytokine, a recombinant cytokine, or a mixture thereof. The dosage of the cytokine contained in the ODT is not particularly limited and can be determined according to the situation.

[0013] In some embodiments, the cytokine is selected from an interferon, such as type I INF, type II INF, and type III INF, etc. In one embodiment, the cytokine is interferon-alpha (INF-α), more preferably recombinant INF-α.

[0014] In one embodiment, the ODT may contain a dosage of 1-100,000 international units (IU) of INF-α. In one embodiment, the ODT may contain a dosage of 1,000 micrograms or less of INF-α. The dosage may be, for example, less than 75,000 IU, less than 50,000 IU, less than 25,000 IU, less than 10,000 IU, less than 7,500 IU, less than 5,000 IU, less than 2,500 IU, less than 1,000 IU, more than 10 IU, more than 100 IU, more than 250 IU, more than 500 IU, more than 750 IU, more than 900 IU, or a value between any two of the above values.

[0015] In the present application, the ODT includes at least one excipient. The excipient is a pharma- ceutically acceptable ingredient, and any known excipient can be used as long as it can function as an excipient in the ODT and does not adversely affect the cytokine. In an embodiment, the excipient may include, for example, sugars, sugar alcohols, cellulose, polysaccharides, stearic acid, magnesium stearate, calcium stearate, polyethylene glycol, sodium lauryl sulfate, glycerin, anhydrous dicalcium phosphate, magnesium aluminometasilicate, and silicon oxide. Examples of sugars include lactose, maltose, dextrin, trehalose, and sucrose. Examples of sugar alcohols include mannitol, erythritol, isomalt, lactitol, maltitol, sorbitol, and xylitol. Examples of celluloses include crystalline cellulose, silicified crystalline cellulose, and powdered cellulose. Examples of polysaccharides include starches such as corn starch, potato starch, and pregelatinized starch. In some embodiments, the excipient is a sugar. In a preferred embodiment, the excipient is maltose.

[0016] A method for producing an ODT containing a cytokine as an active ingredient includes: acidifying an excipient, performing a first granulation step of the acidified excipient to obtain an acidic powder, and performing a second granulation step by mixing the acidic powder with the cytokine to obtain granules containing the cytokine.

[0017] In the present application, the excipients are acidified before mixing with the active ingredient, i.e., cytokines. The acidified excipients can prevent proteolysis of cytokines during the preparation of ODT and maintain the stability and content of cytokines in the final product.

[0018] In some embodiments, the acidified excipient is prepared by: mixing an acid with water to obtain an acidic solution, and mixing the excipient with the acidic solution. The water can be purified water, sterile water, deionized water, etc.

[0019] The acid is added to adjust the pH of the excipient solution. In some embodiments, the acidification causes the excipient solution to have a pH of 3 to 10, such as a pH of 3.5 to 8, a pH of 4 to 7, or a pH of 5 to 6. Inorganic and / or organic acids can be used as long as they do not destroy or change the components or properties of the ODT. Examples of acids include, but are not limited to, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, carbonic acid, acetic acid, citric acid, malic acid, succinic acid, fumaric acid, and lactic acid. In some embodiments, the acid is citric acid.

[0020] The granulation step in the present application is not limited in any way, and can be applied with conventional granulation methods. Examples of granulation methods include high-speed shear mixing granulation, fluidized bed granulation, centrifugal granulation, roller granulation, and spray granulation. Considering the characteristics of the active ingredient contained in ODT, wet granulation method is preferred. In some embodiments, high-speed shear mixing granulation is applied. In some embodiments, fluidized bed granulation is applied.

[0021] After the excipient acidification step and the first granulation step, the obtained acidic powder is further processed. Hereinafter, the acidic powder obtained from the first granulation step may be referred to as the "first acidic powder". The first acidic powder may be sieved, for example, by using a 100 mesh screen, and the powder that passes through the screen (hereinafter referred to as the "sieved portion of the first acidic powder" or "undersized material") and the powder that remains on the screen (hereinafter referred to as the "remaining portion of the first acidic powder") are separated. The sieved portion of the first acidic powder is further mixed with non-acidified excipients and then granulated to obtain a second acidic powder.

[0022] The acidic powder can then be mixed with the cytokines.

[0023] The cytokine may be contained in a solution. The solution may further include a solvent and an acid. In one embodiment, the solution including the cytokine, the solvent, and the acid has a pH of 3 to 10, such as a pH of 3.5 to 8, a pH of 4 to 7, or a pH of 5 to 6. The solvent may be any pharma- ceutically acceptable solvent, so long as it does not adversely affect the cytokine. In one embodiment, the solvent may be water, such as purified water, deionized water, and / or sterile water. The acid may be an inorganic acid and / or an organic acid, so long as it does not adversely affect the cytokine. Examples of acids may include, but are not limited to, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, carbonic acid, acetic acid, citric acid, malic acid, succinic acid, fumaric acid, and lactic acid. In some embodiments, the acid is hydrochloric acid (HCl). In some embodiments, the solution further includes an excipient, which may be the same as or different from the excipient that underwent acidification. In some embodiments, the solution may further comprise an additive selected from the group including a buffering agent, a disintegrant, a binder, a coloring agent, a lubricant, a flow agent, a surfactant, a flavoring agent, an antioxidant, a preservative, or any combination thereof.

[0024] In some embodiments, a first acidic powder is mixed with a cytokine and a second granulation step is performed. In some embodiments, a second acidic powder is mixed with a cytokine and a second granulation step is performed.

[0025] In some preferred embodiments, a second acidic powder is mixed with the cytokine and a second granulation step is performed to obtain a first granule, and then the first granule is further mixed with the remaining portion of the first acidic powder and a third granulation step is performed to obtain a second granule, or a compression step is performed to obtain a tablet.

[0026] In some embodiments, one or more additional excipients can be added at any suitable step of the method.In some embodiments, additives such as buffers, disintegrants, binders, colorants, lubricants, flow agents, surfactants, flavoring agents, antioxidants, preservatives, fillers, or any combination thereof can be added at any suitable step of the method.Such additives are not particularly limited and can be applied or added based on the general knowledge in the relevant technical field.In one embodiment, the additive is magnesium stearate.

[0027] In accordance with the above, the present application provides, in one embodiment, a method for producing an ODT comprising a cytokine, the method comprising: acidifying a first excipient; performing a first granulation step of the acidified first excipient to obtain a first acidic powder; sieving and granulating the first acidic powder to obtain a second acidic powder; and performing a second granulation step by mixing the second acidic powder with the cytokine to obtain first granules.

[0028] The present application also provides, in one embodiment, a method for producing an ODT comprising a cytokine, the method comprising: acidifying a first excipient; performing a first granulation step of the acidified first excipient to obtain a first acidic powder; sieving and granulating the first acidic powder to obtain a second acidic powder; performing a second granulation step by mixing the second acidic powder with the cytokine to obtain first granules; and compressing the first granules to form a tablet.

[0029] Furthermore, in one embodiment, the present application also provides a method for preparing an ODT comprising a cytokine as an active ingredient, the method comprising: acidifying a first excipient, performing a first granulation step of the acidified first excipient to obtain a first acidic powder, sieving and granulating the first acidic powder to obtain a second acidic powder, performing a second granulation step by mixing the second acidic powder with the cytokine to obtain a first granule, and mixing the first granule with the first excipient and / or additive and compressing to form a tablet. In particular, the remaining portion of the first acidic powder can be mixed with the first granule.

[0030] In this application, the method may further include a drying step, a sieving step, and a compression step. The drying step may be performed after the first granulation step, the second granulation step, and / or the third granulation step. After the drying step, a sieving step may be optionally performed to improve the uniformity of the resulting granules. The mesh size of the screen used in the sieving step may be determined according to need. After all the granulation steps are completed, a compression step may be performed to compress the granules containing cytokines to form tablets.

[0031] In another aspect, the present application also provides an ODT prepared by the method described herein. The ODT can maintain good and sufficient efficacy and stability of the cytokines contained therein.

[0032] The present application provides a method for producing cytokine-containing ODTs with optimized processing steps suitable for industrial-scale production. The method can be applied to produce cytokine-containing ODTs with good protein stability and cytokine potency. EXAMPLES

[0033] Referring to Figure 1, the steps of the process for producing an ODT containing IFN-α are illustrated. In this example, maltose was used as the excipient to be acidified.

[0034] Preparation of adjusted activity bulk solution: HCl for pH adjustment, sodium acetate trihydrate, and sodium chloride (NaCl) were added to purified water. Stirring was continued for at least 10 minutes until a clear solution was obtained by visual observation. Human serum albumin was added and stirred for 10 minutes. INF-α solution was slowly added to this solution and stirring was continued for 10 minutes. As such, the adjusted active bulk solution was obtained.

[0035] Preparation of acid powder: Maltose was ground and passed through a No. 40 mesh. The maltose powder was sieved through a Comil (screen openings 0.032 inches, 032R).

[0036] Citric acid was slowly added to the purified water with stirring. Stirring was continued for 10 minutes to obtain a clear appearing acidic solution.

[0037] The acidic solution and maltose powder were mixed and fed into a high speed mixer granulator to produce an acidic powder. The acidic powder was dried by using a fluid bed granulator dryer / coater. Optionally, after the drying step and before the treatment step, the acidic powder was passed through a 30 mesh screen (vibro sieve separator). As such, a first acidic powder was obtained.

[0038] Acid powder treatment: The first acidic powder was sieved by using a 100 mesh screen (vibro sieve separator) to separate the undersized material (the acidic powder that passes through the screen) from the remaining portion of the first acidic powder (the acidic powder that does not pass through the screen).

[0039] The undersize material can be mixed with maltose powder in a ratio of 10-50 wt% to 90-50 wt% of the undersize material. In this Example 1, the undersize material is 20%. This mixture is fed into a high speed mixer granulator to obtain a second acidic powder.

[0040] Preparation of granules containing INF-α: The second acidic powder was mixed with the adjusted active bulk solution. This mixture was fed into a high-speed mixer granulator, and a binder-in and granulation step was carried out to obtain a first granule containing INF-α. Then, the first granule was dried at 30°C for 15 minutes by using a fluidized bed granulator dryer / coater, and sieved through a 60 mesh screen (vibrosieve separator).

[0041] The first granules were mixed with the remaining portion of the first acidic powder in a drum mixer for 10 minutes. Magnesium stearate was added as an additional excipient and mixed in a drum mixer for 5 minutes.

[0042] A high speed mixer granulator was applied to prepare the second granule containing INF-α.

[0043] In an alternative embodiment, magnesium stearate could be added before the mixture was subjected to tablet compression.

[0044] In the following examples, various parameters of the process were varied and evaluated to find optimal steps / parameters / conditions for the process. EXAMPLES

[0045] Evaluation of various factors for protection against INF-α: The experiment was to find the optimal solution for preparing the actively bulk solution, which is very important for the potency of the API. The optimal solution could preserve the potency of the API throughout the manufacturing process. Table 1 shows the two-factor experimental design.

[0046] [Table 1] Figure 2 shows that for the protective effects of various factors on INF-α, positive values ​​have a protective effect, negative values ​​have a destructive effect on INF-α, and there is an interaction between the two factors, with positive values ​​strengthening the protective effect of the interaction and negative values ​​weakening the protective effect.

[0047] result All cytokines need to stay in the buffer system with surfactants / stabilizers when dissolved, but not with stressing agents. Most biological medicines are blood products, serum products, vaccines, cell therapy products, gene therapy products, biotechnology products, human or animal extract products, allergen products, tissue engineering products. All the above biological medicines are sterile products. However, the cytokine-containing ODT produced by the method of the present application is a non-sterile product. The first objective was how to protect and maintain the efficacy and function of INF-α contained in ODT. The inventors not only set up a more suitable manufacturing process, but also found the optimal parameters for API. According to the inventors' research, referring to FIG. 2, HS (human serum albumin), NaCl, pH adjustment, WFI (water for injection), and EDTA have a protective effect. In contrast, the higher the IFN concentration, the lower the protective effect. It can be explained that the higher the IFN concentration, the lower the HS protective effect. In addition, salt and HS are antagonistic to each other, with the higher concentrations of both reducing the protective effect, and the other experimental factors interacting with each other more strongly. This knowledge, combined with the two manufacturing methods, suggests that biological solid dosage forms have beneficial effects. EXAMPLES

[0048] Evaluation of the excipient acidification step: In this example, a method for preparing a combination containing INF-α and maltose was demonstrated. First, 1 kg of maltose powder was acidified using 0.1% citric acid solution in a high shear mixer (SMG), then granulated and dried in a fluid bed granulator dryer (FBD) for 15 minutes to obtain an acidified powder. Next, 8.9 μg of API solution, 510 mg of sodium acetate, 2.76 g of NaCl, 450 mg of HSA, and 1.65 ml of HCl were mixed with sterile water or pure water to prepare 150 ml of active bulk solution. Then, 472.4 g of acidified powder and the active bulk solution were granulated and dried using FBD. The content of INF-α in the second granulated powder was measured by ELISA, and a higher retention rate was revealed in the acidified group when comparing the acidified group with the non-acidified group, as shown in Figure 3, indicating that the excipient acidification step can effectively protect the stability of IFN-α. EXAMPLES

[0049] Percentage of water in the acidifying solution: In this experiment, adjustment of the percentage of water in the acidification solution was tested to evaluate its impact on the granulation step. 0.5 kg of maltose powder was tested with different amounts of colored water. A more uniform particle distribution was observed when the water / maltose ratio ranged from 6 to 8 g / 100 g. However, when the water content was too low (5 g / 100 g), coarse and fine particles of different colors were observed after granulation, indicating a poor powder uniformity. On the other hand, excessive water content led to the dissolution of maltose and its adhesion to the walls of the container, resulting in the formation of large crystal aggregates that had a negative effect on the yield. EXAMPLES

[0050] Acidification granulation equipment: This study utilized 0.5 kg of maltose powder and 0.1% citric acid solution along with the tested equipment for granulation including SMG and FBD. After granulation, the particle size of the powders was examined. It was observed that the FBD group showed a more distinct difference in particle size and showed a significant rate of agglomeration. In contrast, the SMG group showed a more uniform particle size distribution. Therefore, SMG was preferred for granulation of powder forms.

[0051] On the other hand, FBD was preferred for the drying step with a temperature setting of 55-60°C and a drying time of 10-20 minutes. The moisture content analysis showed about 1 ± 0.5%. Similar results could be obtained using a vacuum pump, but it would take several hours. On the other hand, choosing FBD reduced the time cost and the probability of unexpected events during the production process. EXAMPLES

[0052] First acid powder treatment: In this experiment, 1 kg of maltose powder and 0.1% citric acid solution (61 g) were subjected to acidification granulation using SMG and then dried for 15 minutes using FBD. The acidified powder was sieved through a 100 mesh sieve, and 500 g of the undersized particles were mixed with the active bulk solution and granulated / dried using FBD to obtain the second granulated powder. Finally, the second granulated powder: acidified powder: magnesium stearate were mixed in a weight ratio of 1:0.98 to 1:0 to 0.02 to form tablets. The tablets were tested for INF-α potency, and the results were shown in Figure 4. In INF-α-containing ODT, INF-α is required to be uniformly distributed in each tablet. The content of INF-α cannot be directly reflected by the potency test results. As shown in FIG. 4, the sieved group had less than 15% variation in samples, indicating that adding a sieving step prior to the second granulation improves tablet uniformity. EXAMPLES

[0053] Comparison of the second granulator: In this experiment, the equipment tested for the second granulation included SMG and FBD. For FBD, tablets were prepared as in Example 6. For SMG, tablets were prepared as in Example 6, except that the undersize material and active bulk solution were granulated using SMG and then dried using FBD to obtain the second acidic powder. The results are shown in Figure 5 and had a sample-to-sample variability of less than 20%.

[0054] As shown in FIG. 5, granulation with the SMG machine resulted in a higher percentage of IFN adhering to the acidic powder and greater tablet uniformity than granulation with the FBD. EXAMPLES

[0055] Tablet thickness: In this example, the effect of tablet thickness was evaluated. The compression parameters were adjusted to increase the thickness from 3.6 mm to 4.0 mm, and the samples were tested for INF-α potency. The results are shown in Figure 6. The data showed that increasing the tablet thickness did not result in a change in uniformity, but improved the protection of INF-α potency.

[0056] Although the present invention has been disclosed by referring to the above preferred embodiments and examples, it should be understood that these examples are intended in an illustrative and not limiting sense. Modifications and combinations may easily occur to those skilled in the art, and are considered to be within the spirit and scope of the present invention and the scope of the claims, as well as equivalent systems and methods thereof.

Claims

1. 1. A method for producing an orally disintegrating tablet comprising a cytokine as an active ingredient, comprising the steps of: Acidifying the excipient; carrying out a first granulation step of said acidified excipients to obtain an acidic powder; carrying out a second granulation step by mixing the acidic powder with the cytokine to obtain granules containing the cytokine; The method includes:

2. 2. The method of claim 1, wherein the cytokine is selected from the group including chemokines, interferons, interleukins, lymphokines, and tumor necrosis factors.

3. 2. The method of claim 1, wherein the excipient is selected from the group comprising sugars, sugar alcohols, cellulose, polysaccharides, stearic acid, magnesium stearate, calcium stearate, polyethylene glycol, sodium lauryl sulfate, glycerin, anhydrous dicalcium phosphate, magnesium aluminometasilicate, and silicon oxide.

4. The excipient is preparing an acidic solution comprising an acid and water; and mixing the excipient with the acidic solution; It is acidified by 10. The method of claim 1, wherein the resulting solution comprising the excipient, the acid and water has a pH of from 3 to 10.

5. 2. The method of claim 1, wherein the cytokine is contained in a solution comprising a solvent and an additive, and the solution comprising the cytokine, the solvent and the additive has a pH of from 3 to 10.

6. 1. A method for producing an orally disintegrating tablet comprising a cytokine as an active ingredient, comprising the steps of: acidifying the first excipient; conducting a first granulation step of the acidified first excipient to obtain a first acidic powder; sieving and granulating the first acidic powder to obtain a second acidic powder; carrying out a second granulation step by mixing the second acidic powder with the cytokine to obtain a first granule; The method includes:

7. compressing the first granules to form a tablet; or mixing said first granules with said first excipient and / or additive and compressing to form a tablet; The method of claim 6 further comprising:

8. The first excipient is preparing an acidic solution comprising an acid and water; and mixing the excipient with the acidic solution; It is acidified by 7. The method of claim 6, wherein the resulting solution comprising the excipient, the acid and water has a pH of from 3 to 10.

9. 7. The method of claim 6, wherein the cytokine is selected from the group comprising chemokines, interferons, interleukins, lymphokines, and tumor necrosis factors, and the first, second, or third excipient is selected from the group comprising sugars, sugar alcohols, cellulose, polysaccharides, stearic acid, magnesium stearate, calcium stearate, polyethylene glycol, sodium lauryl sulfate, glycerin, anhydrous dicalcium phosphate, magnesium aluminometasilicate, and silicon oxide.

10. 10. An orally disintegrating tablet (ODT) comprising a cytokine as an active ingredient, characterized in that the ODT is produced by the method according to claim 1 or claim 6.

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