Compositions and methods for pretreatment of cancer

Abstract

The present invention relates to a pharmaceutical composition for oral administration, the pharmaceutical composition comprising: a) immediate-release granules comprising i. an active ingredient selected from the group consisting of valproic acid, semi-sodium valproic acid, sodium valproic acid, and magnesium valproic acid, and ii. a filler; and b) sustained-release pellets comprising: i. a pellet core comprising (1) an active ingredient selected from the group consisting of valproic acid, semi-sodium valproic acid, sodium valproic acid, and magnesium valproic acid, and (2) a filler; and ii. on the pellet core iii. A sustained-release pellet comprising a subcoat provided on the subcoat, the content of which is 10 to 20% by weight based on the weight of the pellet core and contains a film-forming agent, and iii. a sustained-release coating provided on the subcoat, the content of which is 25 to 100% by weight based on the weight of the pellet core coated with the subcoat and contains a film-forming agent, wherein the amount of active ingredient in the immediate-release granules accounts for 70 to 80% by weight of the total weight of active ingredients in the pharmaceutical composition, and the amount of active ingredient in the sustained-release pellets accounts for 20 to 30% by weight of the total weight of active ingredients in the pharmaceutical composition. In yet another aspect, the present invention relates to a method for producing a pharmaceutical composition and a pharmaceutical composition for use in a method of pretreatment of cancer.

Description

Technical Field

[0001] Field of the Invention The present invention relates to compositions and methods for pre-treatment of cancer, i.e., administering one or more compositions as pre-treatment prior to other treatments to enhance the effectiveness of treatment or reduce the side effects of treatment for individuals requiring cancer treatment.

Background Art

[0002] Background of the Invention Cancer can be defined as abnormal cell growth that exhibits signs of uncontrolled proliferation and impairment of programmed cell death. From a classical perspective, a series of genetic events cause malignant transformation, resulting in cell clones that do not respect the integrity of other cells and tissues and can ultimately metastasize. Cancer is involved in every tissue of the body and can have many different forms at each site.

[0003] Malignant lymphoma can be defined as the malignant transformation of hematopoietic lymph cells. Lymphomas can be classified into aggressive lymphomas and indolent lymphomas. Aggressive lymphomas are characterized by a rapid growth pattern and can have dramatic clinical features. However, aggressive lymphomas can be cured by chemotherapy, radiotherapy, and treatment with monoclonal antibodies. In contrast, indolent lymphomas (e.g., follicular lymphoma) have a slow growth pattern and usually have milder clinical findings. However, indolent lymphomas cannot be cured by standard lymphoma treatments and may be treatable by allogeneic stem cell transplantation. The median survival period of humans suffering from follicular lymphoma is 8 - 10 years. Diffuse large B-cell lymphoma and Hodgkin lymphoma belong to the group of aggressive lymphomas, and follicular lymphoma and chronic lymphocytic leukemia are indolent lymphomas. Myeloma consists of malignantly transformed plasma cells. These lymphomas are related to indolent lymphomas but are usually considered to exist on their own. The prognosis is pessimistic, and the median survival period is 5 - 7 years.

[0004] One of the most common subtypes of malignant lymphoma is diffuse large B-cell lymphoma (DLBCL), with an incidence of approximately 500 cases per year in Sweden. DLBCL accounts for 60–70% of invasive lymphomas. The median age at diagnosis is 70 years, and DLBCL is slightly more common in men than in women.

[0005] The standard first-line treatment for DLBCL is chemotherapy consisting of a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP). In recent years, the addition of the CD20 antibody rituximab has become the international clinical standard (R-CHOP), improving improved progression-free survival, asymptomatic survival, disease-free survival, and overall survival (Morrison, Expert Rev Anticancer Ther, 2008;8(10):pp.1651-1658). Nevertheless, with 45% of patients still dying from the disease, there is a clear clinical need to increase progression-free survival in DLBCL patients.

[0006] One crucial area in cancer research is the regulation of DNA transcription. This is a complex process, and the mechanisms involved are only partially understood. Histone deacetylases (HDACs) can control the expression of tumor suppressor genes, as well as the activity of transcription factors involved in both cancer development and progression. HDACs act through changes in either DNA or chromatin components via histone deacetylation, affecting the three-dimensional conformation of DNA (epigenetic modification) without altering or interfering with its sequence. It has also been suggested that they may alter sensitivity to DNA-damaging chemotherapy by regulating chromatin structure. From these perspectives, several in vitro studies have suggested that HDAC inhibition may have synergistic effects with chemotherapy.

[0007] Therefore, numerous HDAC inhibitors have been developed in recent years. These can be divided into four classes: hydroxamic acid / carbamic acid, cyclic peptides, fatty acids, and benzamides. Examples of HDAC inhibitors approved for cancer treatment include vorinostat and romidepsin, which are approved by the FDA (Food and Drug Administration) for the treatment of cutaneous T-cell lymphoma and are currently being evaluated for the treatment of other malignancies.

[0008] Clinically, the most well-known HDAC inhibitor is valproic acid, an anticonvulsant that has been used to treat epilepsy since the 1970s. Valproic acid belongs to the fatty acid class of inhibitors.

[0009] The inventors have already shown that HDAC inhibitors combined with steroids are useful when administered as a pretreatment prior to other therapies to people requiring cancer treatment, and that the therapeutic effect is enhanced (see WO2012 / 128709).

[0010] Despite the prospect of more effective cancer treatment resulting from the emergence of prior therapies using HDAC inhibitors as taught in WO2012 / 128709, further development is still needed to fully realize the potential of prior cancer treatment with HDAC inhibitors.

[0011] One of the challenges that arises is how to effectively administer HDAC inhibitors to meet the requirements for their pre-treatment use. Valproic acid, one of the HDAC inhibitors, has been in clinical use since the 1970s, but this clinical use is related to a different medical field, namely the treatment of epilepsy, and not to the new field of pre-cancer treatment. Therefore, previously obtained methods for administering HDAC inhibitors such as valproic acid or its derivatives, and the compositions used in such administrations, do not need to be the most effective when administering HDAC inhibitors such as valproic acid or its derivatives as pre-cancer treatment. [Overview of the project] [Problems that the invention aims to solve]

[0012] Therefore, an object of the present invention is to provide a pharmaceutical composition that provides efficient pretreatment for cancer.

[0013] Another object of the present invention is to provide a pharmaceutical composition that can be manufactured simply and efficiently.

[0014] Another object of the present invention is to provide a method for producing the pharmaceutical composition. Another object of the present invention is to provide a pharmaceutical composition for use in a pretreatment method for cancer.

[0015] A further object of the present invention is to provide a method for pre-treating cancer by administering the pharmaceutical composition. [Means for solving the problem]

[0016] Summary of the Invention According to corresponding first and second aspects of the present invention, at least one of the above objectives is achieved by a pharmaceutical composition for oral administration. The pharmaceutical composition is a) Immediate-release granules, i. An active ingredient selected from the group consisting of valproic acid, semisodium valproate, sodium valproate, and magnesium valproate, ii. Immediate-release granules containing a filler, b) A sustained-release pellet, i. A pellet core, (1) An active ingredient selected from the group consisting of valproic acid, semisodium valproate, sodium valproate, and magnesium valproate, (2) A pellet core containing a filler, ii. A subcoat provided on the pellet core, having a content of 10-20% by weight based on the weight of the pellet core, and containing a film-forming agent, iii. It is provided on the subcoat, and the content is 25 to 100% by weight based on the weight of the pellet core coated with the subcoat, and it includes a sustained-release coating containing a film-forming agent. The amount of the active ingredient in the immediate-release granules accounts for 70 to 80% by weight of the total weight of the active ingredient in the pharmaceutical composition, and the amount of the active ingredient in the sustained-release pellets accounts for 20 to 30% by weight of the total weight of the active ingredient in the pharmaceutical composition. The film-forming agent of the sustained-release coating is different from that of the subcoat. The method for manufacturing the pharmaceutical composition according to the first aspect is as follows. i. The step of manufacturing immediate-release granules. ii. The step of manufacturing pellet cores. iii. The pellet cores are a. The substep of suspending or dissolving the subcoat in an aqueous solvent to form an aqueous solution or dispersion of the subcoat. b. The step of coating with the subcoat by performing the substep of contacting the pellet core with the aqueous solution or dispersion of the subcoat to produce a pellet core provided with the subcoat. iv. The pellet cores provided with the subcoat are a. The substep of suspending or dissolving the sustained-release coating in an aqueous solvent to form an aqueous solution or dispersion of the sustained-release coating. b. The step of further coating with the sustained-release coating by performing the substep of contacting the pellet core coated with the subcoat with the aqueous solution or dispersion of the sustained-release coating to produce a sustained-release pellet.

[0017] Thus, the present invention is based on the discovery that when the subcoat is provided on the pellet core, the subsequent sustained-release coating (which is preferably applied using an aqueous solution or dispersion) does not interfere with the structure of the pellet core, so the release rate of the active ingredient from the sustained-release pellet will not surely become an unexpected rate.

[0018] In other words, as found in Example 1, directly applying a sustained-release coating to the pellet core may lead to an unexpected release rate of the active ingredient. It has been found that this unexpected release rate may be due to the aqueous solution or dispersion used for applying the sustained-release coating. Specifically, it has been found that the combination of the high solubility of the API, the swelling property of MCC, and the porosity of the sustained-release coating allows water to penetrate the coating and reach the pellet core, causing swelling and rupture of the core and rapid release of all the active ingredients.

[0019] This problem of the unexpected release rate was solved by the inventors when a subcoat was applied to the pellet core before applying the sustained-release coating. This subcoat is thought to play a role in protecting the pellet core from the aqueous solution or dispersion, making it possible to apply the sustained-release coating using an aqueous solution or dispersion.

[0020] Furthermore, as shown in Example 2, the pharmaceutical composition according to the first aspect of the present invention has several advantages over known compositions. These advantages include a decrease in the maximum plasma concentration (C max ) of the active ingredient, a reduction in the difference in plasma concentration profiles between postprandial and fasting administrations, a reduction in the risk of accumulation of the active ingredient, and stability of exposure with the maximum concentration of the active ingredient maintained low even when the dosing frequency of the pharmaceutical composition is reduced, such as twice a day. This was different from a reference composition that was thought to show a higher C max when administered twice a day at the corresponding dose.

[0021] Finally, as shown in Example 3, the pharmaceutical composition according to the first aspect of the present invention inhibits histone deacetylation over a wider range than known compositions.

[0022] The third aspect of the present invention relates to a pharmaceutical composition according to the first aspect of the present invention for use in a pre-treatment method for cancer, the pharmaceutical composition being administered to a human suffering from cancer.

Brief Description of the Drawings

[0023] [Figure 1A] This figure shows the dissolution results of a subcoat with a certain content, a sustained-release coating with a certain content, a pore-forming material with a certain content, and sustained-release pellets using a certain drying method. [Figure 1B] This figure shows the dissolution results of a subcoat with a certain content, a sustained-release coating with a certain content, a pore-forming material with a certain content, and sustained-release pellets using a certain drying method. [Figure 1C] This figure shows the dissolution results of a subcoat with a certain content, a sustained-release coating with a certain content, a pore-forming material with a certain content, and sustained-release pellets using a certain drying method. [Figure 1D] This figure shows the dissolution profile of the completed pharmaceutical composition. [Figure 2A] This figure shows the mean (± standard deviation) plasma concentration (μM) of total valproate over nominal time (h). [Figure 2B] This figure shows the data from Figure 2A on a semi-logarithmic graph. [Figure 3A] This figure shows the mean (± standard deviation) plasma concentration (μM) of free (unbound) valproate against nominal time (h). [Figure 3B] This figure shows the data from Figure 3A on a semi-logarithmic graph. [Figure 4A] This figure shows the mean (± standard deviation) plasma concentration (μM) of total valproate as a percentage of nominal time (h) when 30 mg of valproate per kg of body weight is administered twice daily. [Figure 4B] This figure shows the mean (± standard deviation) plasma concentration (μM) of free valproate as a percentage of nominal time (h) after administering 20 mg of valproate per kg of body weight three times a day. [Figure 5] This box plot shows the acetylation of lysine H3 to lysine 9 (acH3K9) in peripheral blood mononuclear cells (PBMCs) compared to baseline expression. [Modes for carrying out the invention]

[0024] Detailed explanation Corresponding first and second aspects of the present invention relate to pharmaceutical compositions for oral administration, wherein the pharmaceutical composition is: a) Immediate-release granules, i. An active ingredient selected from the group consisting of valproic acid, semisodium valproate, sodium valproate, and magnesium valproate, ii. Immediate-release granules containing a filler, b) A sustained-release pellet, i. A pellet core, (1) An active ingredient selected from the group consisting of valproic acid, semisodium valproate, sodium valproate, and magnesium valproate, (2) A pellet core containing a filler, ii. A subcoat provided on the pellet core, with a content of 10-20% by weight based on the weight of the pellet core, containing a film-forming agent, iii. A sustained-release pellet comprising a sustained-release coating provided on a subcoat, the content of which is 25 to 100% by weight based on the weight of the pellet core coated with the subcoat, and which contains a film-forming agent, The amount of active ingredient in immediate-release granules accounts for 70-80% by weight of the total weight of active ingredients in the pharmaceutical composition, and the amount of active ingredient in sustained-release pellets accounts for 20-30% by weight of the total weight of active ingredients in the pharmaceutical composition. Unlike the film-forming agents used in subcoats, the film-forming agents in sustained-release coatings are different. A method for producing a pharmaceutical composition according to the first embodiment is: i. Steps for producing immediate-release granules, ii. Steps for manufacturing the pellet core, iii. The pellet core, a. A substep of suspending or dissolving the subcoat in an aqueous solvent to form an aqueous solution or dispersion of the subcoat, b. A step of coating with a subcoat by performing a substep of bringing a pellet core into contact with an aqueous solution or dispersion of the subcoat to produce a pellet core with a subcoat, iv. A pellet core with a subcoat is provided. a. A substep of suspending or dissolving the sustained-release coating in an aqueous solvent to form an aqueous solution or dispersion of the sustained-release coating, The method comprises a substep of further coating a pellet core coated with subcoat b with an aqueous solution or dispersion of a sustained-release coating to produce sustained-release pellets, and a substep of further coating with a sustained-release coating.

[0025] The pharmaceutical composition is for oral administration, i.e., suitable for oral administration. In other words, the pharmaceutical composition is formulated to be taken orally by the subject. The pharmaceutical composition can be made suitable for oral administration by packaging the immediate-release granules and sustained-release pellets so that they can be swallowed or ingested by the subject. The pharmaceutical composition may include a capsule containing immediate-release granules and sustained-release pellets. The capsule may be of swallowable size and may be made of a material that dissolves in the stomach or intestines to release the immediate-release granules and sustained-release pellets. As an alternative to a capsule, the pharmaceutical composition may include a pouch or bag that holds the immediate-release granules and sustained-release pellets. The pouch or bag may be made of a material that dissolves in the stomach or intestines to release the immediate-release granules and sustained-release pellets. Alternatively, the capsule, pouch or bag may be opened to release the immediate-release granules and sustained-release pellets, which are then ingested directly.

[0026] The immediate-release granules and sustained-release pellets may be packaged in separate capsules, pouches, or bags, and each may be swallowed or ingested, or opened as described above to release the immediate-release granules and sustained-release pellets.

[0027] The pharmaceutical composition is suitable for pretreatment of cancer, and the cancer is preferably selected from the group consisting of diffuse large B cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia, T-cell lymphoma, myeloma, and Hodgkin lymphoma.

[0028] Immediate-release granules are granules configured to release the active ingredient immediately, or at least immediately after ingestion. This allows for a rapid or abrupt release of the active ingredient, which in turn allows for rapid uptake into the target bloodstream. This reduces the time it takes for the active ingredient to reach an effective concentration in the target bloodstream after administration of the pharmaceutical composition.

[0029] Preferably, the immediate-release granules release at least 75% by weight, for example at least 80% by weight, preferably at least 95% by weight, for example 100% by weight, of the active ingredient contained in the granules within 15 minutes in a pH 1 solvent.

[0030] The release test of the active ingredient in a pH 1 solvent was performed by placing immediate-release granules in a basket and lowering the basket into a 0.1N HCl solvent. Samples were taken at 5, 15, 30, 45, and 60 minutes, and the amount of active ingredient released from the immediate-release granules was measured. During the test, the solvent was stirred using a paddle or by rotating the basket. The stirring speed (paddle or basket) was 100 rpm.

[0031] The number of immediate-release granules in a pharmaceutical composition is determined by the amount of active ingredient in each granule, the total amount of active ingredient in the pharmaceutical composition, and the ratio of the amount of active ingredient in the immediate-release granules to the amount of active ingredient in the sustained-release pellets. Typically, a pharmaceutical composition contains multiple immediate-release granules. Immediate-release granules are usually not spherical.

[0032] The active ingredient is selected from the group consisting of valproic acid, semisodium valproate, sodium valproate, and magnesium valproate.

[0033] Valproic acid is a histone deacetylation inhibitor, or HDAC inhibitor. As shown in WO2012 / 128709, HDAC inhibitors are useful when administered as a precursor therapy to other treatments to patients requiring cancer treatment, particularly in combination with steroids.

[0034] Chemically, valproic acid (VPA) is an organic weak acid. Its conjugate base is valproate. The sodium salt of this acid is sodium valproate, and the magnesium salt is magnesium valproate. The coordination complex of valproic acid and sodium valproate is known as semisodium valproate. The latter is sold under various trade names: Depakote®, Depakote ER, Depakene®, Depacon®, Depakine®, Valparin®, Stavzor®, and Ergenyl®. Sodium valproate is sold in Sweden as Absenor®, Depakine, and Orfiril®. Valproic acid is sold in Sweden as Ergenyl and Depakine.

[0035] The filler aims to provide a base material to which the active ingredient can be mixed. The filler further contributes to providing a compressible mass from which ready-release granules can be produced. The filler may contain, or be composed of, for example, cellulose (preferably microcrystalline cellulose, etc.) and / or calcium phosphate.

[0036] Sustained-release pellets are pellets designed to release the active ingredient over a sustained-release period.

[0037] Preferably, the sustained-release pellets release 95-100%, preferably 100%, of the active ingredients within 8 hours at pH 6.8.

[0038] Furthermore, it is desirable that the sustained-release pellets release 55-65%, preferably 60%, of the active ingredients within 3 hours at pH 6.8.

[0039] Furthermore, it is desirable that the sustained-release pellets release 25-35%, preferably 30%, of the active ingredients they contain within 2 hours.

[0040] The release of the active ingredient in a pH 6.8 solvent was tested by placing a 100 mg sustained-release pellet in a basket, immersing the basket in 0.1 N HCl solvent for 15 minutes, and then removing it. The container holding the pH 1 solvent was washed and filled with a pH 6.8 phosphate buffer solution equilibrated at 37°C. Samples were taken at 30 minutes, 1 hour, 2 hours, 3 hours, 5 hours, 8 hours, and 12 hours. During the test, the solvent was stirred using a paddle or by rotating the basket. The stirring speed (paddle or basket) was maintained at 100 rpm throughout the test, and the test was finally terminated by continuous rotation at 250 rpm for 15 minutes.

[0041] The term "sustained-release" is understood to encompass the term "sustained-release." In the context of this invention, the terms "granules" and "pellets" are used to distinguish between the immediate-release portion and the sustained-release portion of a pharmaceutical composition. Therefore, sustained-release pellets may have the same size and shape as granules and may be formed by the same process. However, generally, immediate-release granules are smaller than sustained-release pellets. Furthermore, immediate-release granules may have a lower degree of sphericity than sustained-release pellets. Moreover, immediate-release granules may be formed by a different process than sustained-release pellets.

[0042] The pellet core is the innermost part of a sustained-release pellet. Because the pellet core is formed from the active ingredient and filler, it can generally release the active ingredient rapidly or immediately. The pellet core becomes a sustained-release pellet mainly by applying a sustained-release coating, because the sustained-release coating slows down the release of the active ingredient from the pellet core.

[0043] As described above, the active ingredients in the pellet core are selected from the same active ingredients as those contained in the immediate-release granules. Preferably, the active ingredients are the same in both the immediate-release granules and the pellet core of the sustained-release pellets, but they may be different. Furthermore, the active ingredients may include mixtures of the described forms of valproic acid, and the pellet cores of the immediate-release granules and the sustained-release pellets may contain different active ingredients or mixtures of different active ingredients.

[0044] However, preferably, in both the immediate-release granules and the pellet core of the sustained-release pellets, the active ingredient is sodium valproate.

[0045] Similar to the fillers in immediate-release granules, the fillers in pellet cores serve the purpose of providing a base material to which active ingredients can be mixed. The fillers further help to provide a mass suitable for processing into which pellet cores can be manufactured. The fillers may contain, or be composed of, for example, cellulose (preferably microcrystalline cellulose) and / or calcium phosphate.

[0046] The subcoat is provided on the pellet core. That is, the surface of the pellet core is covered at least partially, preferably completely, with the subcoat. Thus, the subcoat is applied between the pellet core surface and the sustained-release coating. As is evident from the examples, the purpose of the subcoat is to ensure that the sustained-release coating can be reliably applied using an aqueous solution or dispersion. Although not intended to be theoretically bound, the presence of the subcoat is thought to prevent excessive wetting, swelling, and rupture of the pellet core, and the subsequent release of the active ingredients, when the pellet core is brought into contact with the aqueous solution or dispersion containing the sustained-release coating for application. Therefore, the subcoat is particularly configured to limit or reduce wetting of the pellet core during the application of the sustained-release coating.

[0047] By using a subcoat in this way, it becomes possible to apply the sustained-release coating in an aqueous solution or dispersion. Another effect obtained by the subcoat is the smoothing of the pellet core surface. This helps to ensure that the pellet core is well coated with the sustained-release coating because the surface to which the sustained-release coating adheres is smoother. Furthermore, the smoother surface provided by the subcoat allows for the application of the sustained-release coating with a more uniform layer thickness, further improving the uniformity of the sustained-release coating and the release rate of the sustained-release pellets. The subcoat is provided in a content of 10-20% by weight based on the weight of the pellet core. This range has been found to provide the desired protection against wetting of the pellet core while keeping the total content of the subcoat low enough so as not to unnecessarily reduce the active ingredient content in the pellet core and / or unnecessarily increase the size of the sustained-release pellets. The film-forming agent is any agent that, when mixed with an aqueous solution and in contact with the pellet core, forms a film or coating on the pellet core. The film-forming agent may include cellulose such as hydroxypropyl cellulose and / or hydroxypropyl methylcellulose. Preferably, the subcoat and / or film-forming agent is more or less hydrophobic. Hydrophobicity can be increased by increasing the number of aliphatic groups in the film-forming agent, i.e., by substituting the polar hydroxyl groups of cellulose with nonpolar substituents. This is demonstrated by the increase in the number of aliphatic groups in hydroxypropyl cellulose and hydroxypropyl methyl cellulose compared to cellulose. Alternatively, hydrophobicity can be increased by providing a hydrophobic compound or substance such as a mineral like talc (magnesium silicate).

[0048] Preferably, the subcoat contains hydroxypropyl cellulose and / or hydroxypropyl methylcellulose as a film-forming agent. Preferably, the subcoat consists of one or more film-forming agents and one or more hydrophobic compounds or substances.

[0049] The sustained-release coating is provided on the subcoat. This means that the sustained-release coating covers the subcoat, and thereby the pellet core, at least partially, preferably completely. The purpose of the sustained-release coating is to dissolve gradually in the small intestine so that the active ingredient is released slowly from the sustained-release pellet. This ensures that the active ingredient is released steadily and slowly, and as a result, is absorbed steadily and slowly into the target bloodstream. This helps to maintain the concentration of the active ingredient in the bloodstream above the effective concentration during the administration interval of the pharmaceutical composition.

[0050] The sustained-release coating is provided in a content of 25-100% by weight, based on the weight of the pellet core coated with the subcoat. This range has been found to provide the desired release profile for the sustained-release pellets. The film-forming agent may be any agent that, when mixed with an aqueous solution and in contact with the pellet core coated with the subcoat, forms a film or coating on the subcoat. For example, the film-forming agent may contain cellulose such as ethylcellulose. The film-forming agent of the sustained-release coating is different from the film-forming agent of the subcoat. Preferably, the film-forming agent of the sustained-release coating has higher hydrophobicity, i.e., a higher content of aliphatic groups, compared to the film-forming agent of the subcoat. As an example, the sustained-release coating preferably contains ethylcellulose having a higher content of aliphatic groups compared to the preferred hydroxypropylcellulose and hydroxypropylmethylcellulose in the subcoat. The film-forming agent of the sustained-release coating is defined as being different from the film-forming agent of the subcoat. In other words, the subcoat is chemically different from the sustained-release coating, for example, by containing different compounds. In yet another way of putting it, the coating is not a layer of sustained-release coating, such as the first layer of sustained-release coating applied to the pellet core. In yet another way of putting it, the sustained-release coating is not a layer of subcoats, such as the second layer of subcoat applied to the first layer of subcoat (the first layer is applied to the pellet core). In yet another way of putting it, the subcoat and the sustained-release coating are neither two layers of subcoat nor two layers of sustained-release coating.

[0051] The sustained-release coating may further contain 0-10%, preferably 0-5%, more preferably 1-4% (e.g., 2-3%) of pore-forming material. By supplying pore-forming material to the sustained-release coating, it is possible to further adjust the release rate of the active ingredient from the sustained-release pellets.

[0052] Pore-forming agents are agents that are more hydrophilic than the film-forming agents in sustained-release coatings. When the pore-forming agent is dispersed in the sustained-release coating and comes into contact with an aqueous solvent, it forms pores in the coating due to its high hydrophilicity.

[0053] The sustained-release coating preferably contains ethylcellulose as a film-forming agent. The sustained-release coating preferably consists of one or more film-forming agents and optionally one or more pore-forming agents.

[0054] The amount of active ingredient in the immediate-release granules accounts for 70-80% by weight of the total weight of active ingredients in the pharmaceutical composition, while the amount of active ingredient in the sustained-release pellets accounts for 20-30% by weight of the total weight of active ingredients in the pharmaceutical composition. The sum of these percentages is 100% by weight. In other words, 70-80% by weight of the total weight of active ingredients in the pharmaceutical composition is present in the immediate-release granules, and the remaining active ingredient is present in the sustained-release pellets.

[0055] The inventors have discovered that by distributing the active ingredient in these immediate-release granules and sustained-release pellets, a good balance is achieved between rapidly reaching an effective concentration of the active ingredient in the bloodstream and maintaining the concentration of the active ingredient above the effective concentration during the administration interval of the pharmaceutical composition.

[0056] Step i of the production of immediate-release granules may include a substep of mixing the active ingredient with a filler, followed by a substep of compressing the mixture to form granules. Compression is preferably carried out by slugging and grinding, and more preferably by roller compression. The resulting granules may be further dried.

[0057] Step ii of manufacturing the pellet core may include a substep of mixing the active ingredients with the filler, followed by a substep of compressing the mixture to form pellets. The pellets may be manufactured in particular by extrusion and spheroidization. The resulting pellets are preferably dried. Preferably, the pellets are manufactured and / or dried so that the moisture content is less than 5%, preferably less than 3.5%.

[0058] Step iii... of coating the pellet core with a subcoat is performed on the manufactured pellet core. The coating step includes a substep of suspending or dissolving the subcoat in an aqueous solvent to form an aqueous solution or dispersion of the subcoat, followed by a substep of contacting the pellet core with the aqueous solution or dispersion of the subcoat. Contact may be performed by mixing the pellet core with the aqueous solution or dispersion of the subcoat. After contact, a drying step may be performed to dry the pellet core. By coating the pellet core with the subcoat, a pellet core with a subcoat is obtained. Step iv... of further coating the pellet core with a sustained-release coating is performed, including a substep of suspending or dissolving the sustained-release coating in an aqueous solvent to form an aqueous solution or dispersion, and a substep of contacting the pellet core with the subcoat with the aqueous solution or dispersion of the sustained-release coating to produce a sustained-release pellet. As described above, contact may be performed by mixing the pellet core with the subcoat with the aqueous solution or dispersion of the sustained-release coating. The aqueous solvent preferably contains at least 90%, preferably at least 95%, and more preferably 99-100% water. Typically, the aqueous solution or dispersion is an aqueous dispersion.

[0059] After step iv, it is preferable to dry the sustained-release pellets until the moisture content is less than 5%, preferably less than 3.5%. This may be done in a fluidized bed at 55-65°C, preferably 50-60°C for 60-150 minutes, preferably 80-150 minutes, more preferably 90-150 minutes. However, it is more preferable to dry the sustained-release pellets in an oven at 35-45°C, preferably 40°C for 18-30 hours, preferably 22-26 hours (e.g., 24 hours).

[0060] Generally, sustained-release pellets do not need to have an enteric coating in addition to the sustained-release coating.

[0061] More preferably, the sustained-release pellets after manufacturing should not be re-wetted with an aqueous solvent. This is because the active drug component may migrate through the sustained-release coating.

[0062] The amount of active ingredient in immediate-release granules is preferably 72 to 77% by weight, preferably 75% by weight, of the total weight of active ingredients in the pharmaceutical composition, and the amount of active ingredient in sustained-release pellets is preferably 23 to 27% by weight, preferably 25% by weight, of the total weight of active ingredients in the pharmaceutical composition.

[0063] These ranges further reduce the time from administration of the pharmaceutical composition to reaching effective plasma concentration. Furthermore, these ranges further reduce the maximum plasma concentration.

[0064] Preferably, the maximum diameter of the immediate-release granules is 710 to 1000 μm, and the maximum diameter of the sustained-release pellets is 1250 to 1700 μm. Here, the maximum diameter corresponds to the maximum dimensions such as length, width, and height of the immediate-release granules and sustained-release pellets.

[0065] This maximum diameter range facilitates the passage of granules through the stomach in a manner similar to that of a liquid, that is, without being obstructed by the action of the pyloric sphincter.

[0066] Immediate-release granules and sustained-release pellets may be manufactured using sieves to have their maximum diameters. A first sieve with an opening corresponding to the upper limit of these ranges can be used to selectively remove immediate-release granules and sustained-release pellets having a maximum diameter exceeding this upper limit, respectively. A second sieve with an opening corresponding to the lower limit of these ranges can be used to remove immediate-release granules and sustained-release pellets having a maximum diameter below this lower limit, respectively.

[0067] The active ingredient content in the immediate-release granules is 75 to 95% by weight, preferably 77 to 83% by weight, based on the weight of the immediate-release granules, and the active ingredient content in the pellet core is 35 to 45% by weight, more preferably 37 to 43% by weight, based on the weight of the pellet core.

[0068] This has the advantage of reducing the amount of pharmaceutical composition that must be administered to obtain an effective concentration, because it contains a higher amount of the active ingredient compared to other components such as fillers and other excipients.

[0069] These ratios and ranges have further been shown to provide a good balance in maintaining a high content of active ingredients in the pharmaceutical composition while maintaining good compressibility and moldability during the production of immediate-release granules and sustained-release pellets.

[0070] Preferably, the filler content in the immediate-release granules is at least 5% by weight based on the weight of the immediate-release granules, and the filler content in the pellet core is at least 50% by weight based on the weight of the pellet core.

[0071] These filler values ​​are preferable in that they contribute to maintaining good compressibility and moldability during the manufacturing process of immediate-release granules and sustained-release pellets.

[0072] Preferably, the filler in the immediate-release granules is dicalcium phosphate, and the filler in the pellet core is microcrystalline cellulose.

[0073] It is clear that dicalcium phosphate is an effective filler in immediate-release granules, which generally have a low filler content. In contrast, microcrystalline cellulose is clearly an effective filler in pellet cores, where the filler content is high and the wettability of microcrystalline cellulose can contribute to controlling the release rate of active ingredients during the dissolution process of sustained-release pellets.

[0074] Preferably, the immediate-release granules further contain 0.3 to 1% by weight, preferably 0.5% by weight, of the lubricant based on the weight of the immediate-release granules, and the pellet core further contains 0.5 to 2% by weight, preferably 1% by weight, of the lubricant based on the weight of the pellet core.

[0075] Adding these amounts of lubricant further facilitates the production of immediate-release granules and sustained-release pellets.

[0076] In either case, magnesium stearate is preferred as the lubricant. Preferably, the subcoat content in the sustained-release pellet is 14 to 18% by weight, preferably 15% by weight, based on the weight of the pellet core.

[0077] This amount of subcoat is advantageous because it achieves the objective of preventing wetting of the pellet core when applying the sustained-release coating, while being low enough not to interfere with the release rate determination function of the sustained-release coating. The stated content corresponds to coating the pellet core until the corresponding weight increase is obtained.

[0078] Preferably, the subcourt is the weight of the subcourt. i. 35-40% by weight, preferably 37.5% by weight of hydroxypropyl cellulose, ii. 35-40% by weight, preferably 37.5% by weight of hydroxypropyl methylcellulose, and iii. Contains 20-30% by weight, preferably 25% by weight, of talc.

[0079] This combination of ingredients includes added talc (chemical formula Mg3Si4O 10 It has been shown that a subcoat with good hydrophobic properties, partially provided by (OH)2 hydrated magnesium silicate, is obtained, further protecting the pellet core during the application of the sustained-release coating. In these embodiments, hydroxypropyl cellulose and hydroxypropyl methylcellulose are film-forming agents.

[0080] Preferably, the sustained-release coating is provided by the weight of the sustained-release coating. i. 97-100% by weight of a film-forming agent, and ii. A pore-forming material comprising 0-3% by weight of hydroxypropyl methylcellulose or made thereof.

[0081] These ranges enable sustained-release coatings that provide a favorable release profile. Furthermore, a porosity-forming material content of 0-3% by weight contributes to adjusting the release rate from the sustained-release pellets.

[0082] If there are two or more film-forming agents, the 97-100% by weight range for film-forming agents refers to the total content of film-forming agents.

[0083] Preferably, the film-forming agent is ethylcellulose. Preferably, the pore-forming agent contains hydroxypropylmethylcellulose.

[0084] Preferably, the content of the sustained-release coating in the sustained-release pellet is 65 to 80% by weight, preferably 68 to 75% by weight, and more preferably 70% by weight, based on the weight of the pellet core provided with the subcoat.

[0085] These ranges have been shown to favor the release rate of the active ingredient. These amounts of sustained-release coating correspond to the corresponding weight increase when the sustained-release coating is applied to a pellet core coated with a subcoat.

[0086] Preferably, a) Immediate-release granules are, i. 89-91% by weight of valproic acid or valproate, ii. 8.5-10.5% by weight of anhydrous dicalcium phosphate, and iii. Composed of 0.4-0.6% by weight of magnesium stearate, b) The pellet core is i. 39-41% by weight of valproic acid or valproate, ii. 58-60% by weight of microcrystalline cellulose, and iii. Composed of 0.5 to 1.5% by weight of magnesium stearate, c) The sub-court is i. Provided in an amount equivalent to 16-17% of the weight of the pellet core. ii. The weight of the sub-court, 1) 36.5-38.5% by weight of hydroxypropyl cellulose, 2) 36.5-38.5% by weight of hydroxypropyl methylcellulose, and 3) Contains 24-26% by weight of talc, d) The sustained-release coating is i. Provided in an amount equivalent to 69-71% of the weight of the pellet core coated with subcoat. ii. By weight of the sustained-release coating, 1) 97-99% ethylcellulose, and 2) Contains a pore-forming material containing 1-3% hydroxypropyl methylcellulose, The amount of active ingredient in the immediate-release granules accounts for 74-76% by weight of the total weight of the active ingredient in the pharmaceutical composition. The active ingredient content in the sustained-release pellets accounts for 24-26% by weight of the total weight of the active ingredient in the pharmaceutical composition.

[0087] more, a) Immediate-release granules are, i. 90% by weight of valproic acid or valproate, ii. 9.5% by weight of anhydrous dicalcium phosphate, and iii. Composed of 0.5% by weight of magnesium stearate, b) The pellet core is i. 40% by weight of valproic acid or valproate, ii. 59% by weight of microcrystalline cellulose, and iii. Composed of 1% by weight of magnesium stearate, c) The sub-court is i. Provided in an amount equivalent to 15% of the weight of the pellet core, ii. The weight of the sub-court, 1) 37.5% by weight of hydroxypropyl cellulose, 2) 37.5% by weight of hydroxypropyl methylcellulose, and 3) Contains 25% by weight of talc, d) The sustained-release coating is i. Provided in an amount equivalent to 70% of the weight of the pellet core coated with subcoat, ii. By weight of the sustained-release coating, 1) 98% ethylcellulose, and, 2) Contains a pore-forming material containing 2% hydroxypropyl methylcellulose, The amount of active ingredient in the immediate-release granules accounts for 75% by weight of the total weight of active ingredients in the pharmaceutical composition. The amount of active ingredient in the sustained-release pellets accounts for 25% by weight of the total weight of active ingredients in the pharmaceutical composition.

[0088] This embodiment generally corresponds to the formulation obtained in Example 1. Preferably, the pharmaceutical composition further comprises a capsule for containing immediate-release granules and sustained-release pellets, the capsule defining a unit dose of the pharmaceutical composition.

[0089] If the capsule is a size 0 capsule, each unit dose of the pharmaceutical composition may contain 250 to 285 mg of the active ingredient. The capsule is preferably made of a material such as hydroxypropyl methylcellulose that dissolves in the stomach or intestines to release immediate-release granules and sustained-release pellets, as discussed earlier. The amount of active ingredient in each capsule may be selected so that each dose of two daily doses consists of 5 to 12 capsules, for example, 7 to 10 capsules.

[0090] A third aspect of the present invention relates to a pharmaceutical composition according to the first aspect of the present invention, which is used in a pretreatment method for cancer, wherein the pharmaceutical composition is administered to a person suffering from cancer.

[0091] A third aspect of the present invention relates to a pre-treatment method for cancer, comprising administering a pharmaceutical composition according to the first aspect of the present invention to a person suffering from cancer.

[0092] In the context of the present invention, cancer pretreatment includes treatments performed to enhance the sensitivity of the cancer to further treatments. Therefore, cancer pretreatment includes administering one or more compositions to an individual in need of cancer treatment as a pretreatment prior to other treatments to enhance the effect of the treatment or reduce side effects.

[0093] A typical dose of the active ingredient is up to 60 mg per kg of body weight per day, for example, 30 to 60 mg per kg of body weight per day.

[0094] A typical dose of the active ingredient may be 1500-2500 mg per day, more preferably 1800-2200 mg, and most preferably 2000 mg.

[0095] Preferably, the cancer is selected from the group consisting of diffuse large B cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia, T-cell lymphoma, myeloma, and Hodgkin lymphoma.

[0096] As shown in WO2012 / 128709, these cancer types are particularly favorable to prior treatment with active ingredients.

[0097] Preferably, the pharmaceutical composition is administered to a human before treatment with chemotherapy and / or immunotherapy.

[0098] Chemotherapy and / or immunotherapy should be selected based on the type of cancer a person has.

[0099] Chemotherapy may include the administration of CHOP, a combination of cyclophosphamide, doxorubicin, vincristine, and prednisone, which may consist of 750+ / -10% mg / m2 of cyclophosphamide, 50+ / -10% mg / m2 of doxorubicin, 1.4+ / -10% mg / m2 of vincristine, and 50+ / -10% mg / m2 of prednisone. m2 refers to the surface area of ​​a human body.

[0100] Alternatively, chemotherapy may include the administration of R-CHOP, a combination of the antibody rituximab and cyclophosphamide, doxorubicin, vincristine, and prednisone.

[0101] Each of CHOP and R-CHOP may also contain etoposide, and therefore the corresponding chemotherapy regimens are called CHOEP or R-CHOEP.

[0102] Immunotherapy may involve the administration of antibodies, monoclonal antibodies, or functional fragments thereof, such as rituximab, ofatumumab, GA101, tocitumomab, ibritumomab, ocralzumab, vertuzumab, epratuzumab, FTBA05, AME-133V, or R603. All of the antibodies mentioned above bind to CD20 present on B cells. These antibodies may be administered in doses of 375 + / - 10% mg / m2.

[0103] Other treatments that may be performed after prior treatment include surgery, radiation therapy, and gene therapy.

[0104] Preferably, the pharmaceutical composition is administered twice a day. Since the active ingredient is preferably taken with or in connection with food, it is advantageous to administer the pharmaceutical composition twice a day. This increases the likelihood of administration with meals.

[0105] Preferably, a steroid selected from the group consisting of prednisone, prednisolone, dexamethasone, and betamethasone is also administered to humans. The steroid may be administered simultaneously with the pharmaceutical composition or during the administration interval of the pharmaceutical composition.

[0106] Prednisone or prednisolone may be administered in doses of 20 to 200 mg per day, for example, 50 to 200, 100 to 150, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg. This dose may be administered in one dose or in divided doses.

[0107] Betamethasone can be administered in doses ranging from 4 to 32 mg per day, for example, 10-25, 10-20, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or 32 mg. This dose may be administered in one dose or in multiple divided doses.

[0108] Dexamethasone may be administered in doses of 10 to 80 mg per day, for example, 20 to 70, 10, 20, 30, 40, 50, 60, 70, or 80 mg. This dose may be administered in one dose or in divided doses.

[0109] Another aspect of the present invention is -Pharmaceutical compositions in multiple unit doses, -The present invention relates to a kit containing multiple doses of steroids selected from the group consisting of prednisone, prednisolone, dexamethasone, and betamethasone.

[0110] The kit is useful for providing and maintaining an effective concentration of the active ingredient during a single session of pre-cancer therapy. A single session of pre-cancer therapy may include, for example, a period of at least 12 hours (e.g., 24 hours, or at least 36 hours, or at least 48 hours) and less than 96 hours (e.g., less than 84 hours, or less than 72 hours).

[0111] Furthermore, the kit facilitates the implementation of pre-cancer treatment and improves adherence to prior treatment. The kit preferably contains enough unit doses for two or three doses during each 24-hour period of a pre-cancer treatment session. The number of unit doses in the kit may be 2 to 8 doses.

[0112] The pre-cancer treatment session is preferably initiated 48 hours before the start of cancer treatment. Typically, the dose is 6 or 9 units.

[0113] At least one dose may further contain a steroid selected from the group consisting of prednisone, prednisolone, dexamethasone, and betamethasone. In this case, the steroid does not need to be present in each dose.

[0114] In some embodiments of the kit, the number of unit doses is provided in appropriate containers and / or packaging, and the kit preferably further includes instructions regarding the method and timing of administration.

[0115] Therefore, the kit may, for example, include one or more blisters for containing the dosage.

[0116] Preferably, the kit includes a blister pack containing multiple foldable blisters sealed with a breakable seal sheet, each blister containing one or more doses.

[0117] In some embodiments, the kit contains a sufficient number of unit doses to administer multiple sessions (e.g., six sessions) of pre-treatment for cancer. Thus, one kit can be used for all the unit doses required for a typical cancer treatment cycle consisting of six sessions administered at intervals of 14 to 21 days, in which the pharmaceutical composition is administered as pre-treatment prior to other chemotherapy or immunotherapy. [Examples]

[0118] Example 1 - Towards the development of a sustained-release composition of valproate 1.1 Development of immediate-release granules The immediate-release granules were developed to contain the components shown in Table 1.

[0119] [Table 1]

[0120] Immediate-release granules are formed by mixing the materials in a planet blender or high-shear blender. The resulting mixture is roll-compressed to form a solid ribbon, which is then ground through a 1 mm screen to form granules.

[0121] 1.2 Development of Pellet Cores The pellet core was developed to contain the following components (see Table 2).

[0122] [Table 2]

[0123] Pellet cores were manufactured by mixing materials in a planet blender or high-shear blender. Water was added to the mixture, and the resulting wet paste was extruded to form a mass of material, which was then shaped into a pellet core through spheroidization and drying.

[0124] 1.3 Sub-court Development In the initial development stages of sustained-release pellets, pellet cores were coated with various sustained-release aqueous-based coating polymers such as Surelease®, Eudragit® NM30D, and AcryCoat®. These polymers were designed to release over 8-12 hours under basic pH conditions. However, a problem arose because the combination of the highly soluble API in the core and the swelling properties of microcrystalline cellulose in aqueous solvents resulted in the sustained-release coating becoming too porous. This porosity allowed water to permeate the coating and penetrate the core, causing it to swell. This was visually observed as swelling and rupture of the coated core, resulting in the complete release of the API within one hour.

[0125] Various attempts were made to address this problem. Surprisingly, introducing a subcoat yielded an effective reaction that limited the dissolution of the pellet core in acidic solvents. By adding the subcoat at a weight increase of 10% (i.e., 10% by weight relative to the pellet core weight), the sustained-release coating not only performed as intended, but the non-uniformity of the pellet surface was also smoothed out.

[0126] The initial subcoat was applied as an aqueous formulation containing the following components (see Table 3A).

[0127] [Table 3A]

[0128] The subcoat was applied until a 10% weight increase relative to the pellet core weight was obtained. Furthermore, the subcoat was further optimized and its hydrophobicity enhanced by adding talc mineral (magnesium silicate) using hydroxypropyl cellulose and hydroxypropyl methylcellulose. The optimized subcoat contained the following components (see Table 3B).

[0129] [Table 3B]

[0130] An optimized subcoat was provided in an amount equivalent to 14–18%, preferably 15%, of the weight of the pellet core. These amounts of subcoat are particularly advantageous in preventing the active ingredients from migrating from the pellet core and provide a smooth surface for a sustained-release coating.

[0131] 1.4 Development of sustained-release coatings The sustained-release coating was applied as an aqueous formulation containing the following components (see Table 4).

[0132] [Table 4]

[0133] The sustained-release coating was applied until a weight increase of at least 25% by weight was obtained relative to the weight of the subcoated pellet core. Increasing the weight increase of the sustained-release coating from 25% to 35%, 60%, and 100% further reduced the release rate.

[0134] Furthermore, when the concentration of the pore-forming agent was reduced from 20% to 10%, 5%, and 0%, the release rate decreased significantly, with minimal release in acidic solvents and dissolution lasting several hours in phosphate buffer pH 6.8.

[0135] Furthermore, it was confirmed that drying of the sustained-release pellets also affects the release rate. Various pellets were produced. Figures 1A, 1B, and 1C show the dissolution results of sustained-release pellets with varying amounts of sustained-release coating, pore-forming material, and drying / curing methods. Two-step dissolution was used where necessary, with 0.1N HCl in the acidic phase and the aforementioned pH 6.8 phosphate buffer in the alkaline phase to simulate in vitro conditions. Symbols and legends are explained in the table below.

[0136] [Table 5A]

[0137] [Table 5B]

[0138] [Table 5C]

[0139] Two-stage dissolution data for pellets showed that applying a 10% w / w Opadry® subcoat containing a sustained-release Surelease film of up to 60% w / w without porosizing agent resulted in a sustained-release profile exceeding 12 hours, while thinner films (35% w / w) completed release within 6 hours. However, drying / curing the coated pellets at 40°C for 24 hours sometimes resulted in coating smoothing and a significant change in the release profile (35% weight increase, 68% release in 12 hours; 60% weight increase, 31% release in 12 hours). This indicated that drying / curing affected the release profile by agglomerating the film over time, reducing and refining the pores within the film. Pellets coated with 10% and 5% porosizing agent showed faster release rates than those without porosizing agent. Pellets coated with Surelease containing 10% porosity material released 100% within 6 hours after applying a 60% weight increase. A 100% weight increase extended release to over 12 hours, but considering the capsule size limitations, the coated pellets would be excessive. Pellets coated with Surelease containing 5% porosity material released 100% within 2 hours with a 35% weight increase. When coated pellets with a 60% weight increase were oven-dried / cured, they did not release completely even after 12 hours. This data suggests that 10% porosity material may provide a suitable dissolution profile, but the coated pellets are too large (>1.4mm), and that using 5% porosity material with a 60% weight increase, an ideal profile can be achieved with appropriate drying / curing time.

[0140] 1.5 Further Optimization The completed sustained-release pellets were subjected to fluidized bed drying / curing at 55°C for 1 hour and oven drying / curing at 40°C for 24 hours. Significant differences were observed in the release profiles between the oven-dried pellets and the fluidized bed-dried pellets. The oven-dried pellets exhibited a slow release rate over 8 or 12 hours, demonstrating remarkably good performance.

[0141] Samples showing the greatest coating weight increase in each pellet batch (60% or 100%) were further coated with the enteric polymer AcrylEZE® (methacrylic acid copolymer type C). However, the addition of the enteric coating did not further improve the release profile.

[0142] 1.6 Development of Unit-Volume Capsules Immediate-release granules and sustained-release pellets were filled into capsules to obtain a total active ingredient amount of 250 mg or 285 mg. 75% by weight of the active ingredient was supplied by immediate-release granules, and 25% by weight was supplied by sustained-release pellets.

[0143] 1.7 Final Pharmaceutical Composition and Dissolution Test Batch production of the final pharmaceutical composition was carried out. The pharmaceutical composition is as follows:

[0144] Pellet cores as shown in Table 2 were subcoated using the optimized subcoat shown in Table 3B until the weight increased by 15% by weight. The subcoated pellets were further coated (850-1400 μm) using the sustained-release coating shown in Table 4 until the weight increased by 70%, thereby producing sustained-release pellets, which were then filled into capsules.

[0145] The 710-1000 μm immediate-release granules listed in Table 1 were manufactured and the capsules were filled with them.

[0146] When HPMC capsules (size 0EL) were filled with 208.3 mg of immediate-release granules and 301.5 mg of sustained-release pellets, 187.5 mg (75% by weight) of the immediate-release active ingredient and 62.5 mg (25% by weight) of the sustained-release active ingredient were supplied. The total amount of active ingredient in the capsule was 250.0 mg.

[0147] Figure 1D shows the two-stage dissolution profile of the final capsule. As can be seen from the figure, the capsule containing the pharmaceutical composition released rapidly initially (approximately 70% released in 1 hour), and then slowly (approximately 90% released in 4 hours, and approximately 95% released in 6 hours).

[0148] The final capsule samples were further stored for one or three months under various conditions (e.g., various temperatures (5°C, 25°C, and 40°C), humidity (60%, 75% RH)). Storage of the pharmaceutical formulation under these various conditions did not significantly affect its dissolution profile. Further storage tests showed no significant degradation even after 24 months of storage.

[0149] Example 2 - A Phase 1, open-label, randomized trial investigating the pharmacokinetics of pharmaceutical composition VAL001 under fasting and postprandial conditions in healthy subjects. In this study, 25 subjects were administered either the final composition described in 1.7 above, known as VAL001 (N=12), or a reference composition known as Absenor manufactured by Orion Pharma (N=12).

[0150] 2.1 Unit volume (N=12) In the first part of the study, healthy subjects received a single dose of the final pharmaceutical composition (VAL001) described in 1.7 above (equivalent to 30 mg of valproate per kg of body weight) and a single dose of the reference composition (Absenor) in both fasted and postprandial states. Blood samples were then collected at predetermined intervals to measure plasma valproate concentrations. The results are listed in Table 6 below and shown in Figures 2A, 2B, 3A, and 3B.

[0151] [Table 6]

[0152] As shown in the table, the VAL001 composition provides equivalent exposure, but the maximum blood concentration (C) is equivalent. maxThe peak concentration was low. This means that, considering the high concentrations reached at the doses required to achieve the desired effect, the peak concentration was low, and therefore the risk of side effects was low. Thus, given that high doses of valproic acid are generally required to achieve effective pretreatment for cancer, a low peak concentration is particularly advantageous.

[0153] This behavior is also shown in Figures 2A and 3A, along with the corresponding semi-logarithmic graphs, Figures 2B and 3B. Specifically, Figure 2A shows the mean (± standard deviation) plasma concentration (μM) of total valproate against nominal time (h). As is clear from Figures 2A and 2B, the VAL001 composition reacts more rapidly than the reference composition than the reference composition, especially during fasting. max It reaches.

[0154] Correspondingly, Figure 3A shows the mean (± standard deviation) plasma concentration (μM) of free (unbound) valproate against nominal time (h).

[0155] In summary, Table 6 and Figures 2A, 2B, 3A, and 3B show that the VAL001 composition provides an efficient plasma profile when administered with valproate in the case of pre-cancer treatment. In other words, the VAL001 composition is designed to provide a good synergistic effect between valproate pre-treatment and subsequent cancer treatment. max It is particularly suitable for reaching high concentrations of valproate relatively quickly without excessively raising the levels, and for maintaining high concentrations throughout the administration period.

[0156] 2.2 Two doses vs. three doses (N=13) In the second part of the study, the composition / formulation was administered to healthy subjects after feeding either 30 mg of valproate (VAL001) per body weight in two doses, or 20 mg of valproate (Absenor) per body weight in three doses.

[0157] Subsequently, blood samples were taken at designated times to measure the plasma concentration of valproate. The results are listed in Table 7 below and shown in Figures 4A and 4B.

[0158] [Table 7]

[0159] Figure 4A shows the mean (± standard deviation) total plasma concentration (μM) of valproate as a percentage of nominal time (h) when 30 mg of valproate per kg of body weight is administered twice daily for 3 days. On the other hand, Figure 4B shows the mean (± standard deviation) plasma concentration (μM) of free valproate as a percentage of nominal time (h) when 20 mg of valproate per kg of body weight is administered three times daily for 3 days.

[0160] Figure 4A specifically shows that VAL001 provides a more stable concentration profile and exposure of valproate at the same total daily dose, even when administered twice daily instead of three times daily, compared to the reference composition Absenor.

[0161] Similar results are shown in Figure 4B, which further indicates that the VAL001 composition results in less valproate accumulation in plasma compared to the reference Absenor composition.

[0162] Example 3 - Semi-quantitative Western blotting evaluation of acetylated histone (K9) in cytolysate of isolated peripheral blood mononuclear cells. Analysis was conducted by the Institute for Bioanalysis at the University of Turku, Finland, to determine the degree of histone acetylation in the cytolysate of peripheral blood mononuclear cells collected from subjects who had received repeated administration of the final pharmaceutical composition (referred to as VAL001) or the reference composition (Absenor) described in 1.7 above. Histone acetylation was measured by acetylation of lysine 9 of histone H3 (acH3K9) compared to baseline expression.

[0163] Samples were obtained from subjects participating in the clinical trial described in Example 1. PBMC samples were collected three times daily before administration and during the treatment period of Part 2 of the clinical trial. A total of 96 test samples were analyzed, including samples from 24 subjects at four different time points.

[0164] For each sample, the experimental signal of acH3K9 was normalized against the loading control GAPDH to correct for loading variability, and it was confirmed that the observed changes reflected actual differences between samples. The fold change showing the relative acH3K9 expression of the test samples compared to baseline expression before administration was comparable across different blots.

[0165] 3.1 Sample materials and control samples The sample material was a frozen cell pellet of human PBMCs supplied to the bottom of a cryotube stored in an upright position. PBMCs were isolated from whole blood samples using Cell Preparation Tubes® (CPT, BD Biosciences). The cell pellet was washed and diluted with phosphate-buffered saline (PBS) and then diluted to an equivolute cell concentration (0.75–1.0 × 10⁶ per tube). 6 The cells were dispensed into cryotubes to form individual cells. The PBS was discarded, and the cell pellets were stored frozen at a nominal -80°C until they were transported (with dry ice) to the bioanalytical laboratory.

[0166] Prior to the initiation of the investigational treatment, PBMC samples were also collected from each subject. These samples were used as control samples to determine each subject's baseline acH3K9 protein expression. The cryopreserved test samples (each sample divided into three parts (3 × 96 = 288)) were received on dry ice from CRST Oy on September 15, 2022. The test samples were subsequently stored at a nominal -80°C in a freezer at the Bioanalytical Laboratory.

[0167] 3.2 Overview of the experimental treatment The test treatment administered to subjects from whom samples were collected involved repeated administration of either the sustained-release composition or the reference composition according to the present invention.

[0168] 3.3 Method The cell pellet was dissolved in 200 μl of Laemmli sample buffer containing SDS, the thiol reducing agent β-mercaptoethanol, the protease inhibitor (Roche), and the phosphatase inhibitor (Roche). The sample was sonicated for 10 minutes in a TPX tube (Diagenode) using the Diagenode Bioruptor system, heated (95°C, 5 minutes), and centrifuged at high speed (4°C, 5 minutes). The supernatant was separated into a new tube.

[0169] Soluble proteins were separated by molecular weight using SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Before loading, each well of the sample (15 μl of each sample, equivalent to approximately 75,000 cells) was heated at 95°C for 5 minutes and then loaded into Any kD® Mini-PROTEAN® TGX® Precast Protein Gel (Bio-Rad).

[0170] All sample gels analyzed included two lanes of protein references (Precision Plus Protein 2-Color Standards) as molecular weight references. One protein reference lane per gel was displayed overlaid on the ECL channel in the raw data.

[0171] After electrophoresis, proteins were transferred to PVDF membranes using a semi-dry transfer turbo cell (Bio-Rad) with a transfer time of 30 minutes and a constant voltage of 25V, using the "Standard SD" program. The transfer quality of each PVDF membrane was confirmed by Ponceau S staining and visual inspection. After destaining, the membranes were blocked for 1 hour on a plate shaker at room temperature (RT) using a 5% skim milk (Valio) solution in TBST buffer.

[0172] Primary antibody incubation was performed overnight at +4°C on a roller tube. Rabbit anti-acH3K9 antibody (Cell Signaling Technologies) 1:1000 dilution (0.042 μg / ml) and rabbit anti-GAPDH antibody (Abcam) 1:10,000 dilution (0.1 μg / ml) were used. For detection of the primary antibodies, anti-rabbit HRP-conjugated secondary antibody (Cell Signaling Technologies) was used. Specifically, a 1:10,000 dilution (0.006 μg / ml) was used for acH3K9 detection, and a 1:3000 dilution (0.020 μg / ml) was used for GAPDH detection. All antibodies used were diluted with 1% skim milk in TBST buffer.

[0173] The chemiluminescent signal was induced using SuperSignal West Dura Extended Duration Substrate ECL reagent and captured using a Sapphire imaging system (Azure Biosystems) with a CCD camera. The signal was quantified by concentration measurement using Image J version 1.53c, and the analysis lane, signal density plot, and measured peak area were recorded.

[0174] The measured signals (peak areas) were imported into an Excel file. The acH3(K9) signal was normalized against the GAPDH signal, a loading control, to mathematically correct for the unavoidable inter-sample and inter-lane variations in protein loading. The lane normalization coefficient was calculated for each lane by dividing the GAPDH signal value observed in the lane by the highest GAPDH signal observed in the blot. To calculate the normalized experimental signal for acH3K9, the observed experimental signal was divided by the lane normalization coefficient for that lane.

[0175] 3.4 Results The study samples were analyzed using 12 independent 10-well SDS-PAGE gels.

[0176] All Western blots were of good quality and did not require retesting. Reference digital ECL images with protein markers, signal density plots, and measured signals (peak area) were saved and printed as raw data and confirmed to be of visually appropriate quality.

[0177] The acH3K9 signal was normalized relative to the expression of the housekeeping protein GAPDH. Cell / protein loading and antibody dilution were optimized to ensure signal normalization. However, this normalization is assumed to be best performed when the difference between protein loading and GAPDH signaling is small. Therefore, the results should be evaluated in combination with visual examination of ECL images. It should also be noted that normalized acH3K9 signal values ​​are comparable only within samples on the same blot and should not be compared with signals from other blots. However, the reported magnification changes, which show the relative acH3K9 expression of study samples compared to baseline expression before administration, are well comparable between different blots. Relative acH3K9 expression after valproate (VAL001 or Absenor) administration, measured as the mean of magnification changes, was 1.38 (+38% change) at 23.5 hours, 1.49 (+49% change) at 47.5 hours, and 1.12 (+12% change) at 80 hours. The standard deviation (SD) and coefficient of variation (CV) of the change in magnification were 0.50 and 36% at 23.5 hours, 0.64 and 43% at 47.5 hours, and 0.59 and 53% at 80 hours, respectively.

[0178] In the VAL001 treatment group, the mean change in magnification was 1.44 times (+44%) at 23.5 hours, 1.69 times (+69% change) at 47.5 hours, and 1.26 times (+26% change) at 80 hours. The standard deviation (SD) and coefficient of variation (CV%) of the change in magnification were 0.51 and 36% at 23.5 hours, 0.79 and 47% at 47.5 hours, and 0.74 and 59% at 80 hours, respectively.

[0179] In the Absenor treatment group, the mean change in magnification was 1.32 (+32% change) at 23.5 hours, 1.29 (+29% change) at 47.5 hours, and 0.98 (-2% change) at 80 hours. The standard deviation (SD) and coefficient of variation (CV%) of the change in magnification were 0.48 and 36% at 23.5 hours, 0.34 and 26% at 47.5 hours, and 0.33 and 34% at 80 hours, respectively.

[0180] The mean value, percentage change, standard deviation (SD), and coefficient of variation (CV) of the magnification change were calculated in Excel and reported individually for each time point. The standard deviation and CV of the magnification change represent the variability caused by both biological differences between subjects and assay variability, and should be carefully considered when reporting differences between the experimental group and the control sample.

[0181] Figure 5 is a box plot showing the acetylation of lysine 9 on histone H3. 2.5 Discussion As the results show, the final pharmaceutical composition (VAL001) described in 1.7 above resulted in a higher degree of histone acetylation than the reference composition (Absenor), and therefore exhibited a more efficient histone deacetylation inhibitory effect. Accordingly, the pharmaceutical composition according to the present invention makes cancer pretreatment more efficient.

[0182] The present invention is further described by the following items. 1. A pharmaceutical composition for oral administration, a) Immediate-release granules, i. An active ingredient selected from the group consisting of valproic acid, semisodium valproate, sodium valproate, and magnesium valproate, ii. Immediate-release granules containing a filler, b) A sustained-release pellet, i. A pellet core, (1) An active ingredient selected from the group consisting of valproic acid, semisodium valproate, sodium valproate, and magnesium valproate, (2) A pellet core containing a filler, ii. A subcoat provided on the pellet core, with a content of 10-20% by weight based on the weight of the pellet core, containing a film-forming agent, iii. A sustained-release pellet comprising a sustained-release coating provided on a subcoat, the content of which is 25 to 100% by weight based on the weight of the pellet core coated with the subcoat, and which contains a film-forming agent, A pharmaceutical composition in which the amount of active ingredient in immediate-release granules accounts for 70-80% by weight of the total weight of active ingredients in the pharmaceutical composition, and the amount of active ingredient in sustained-release pellets accounts for 20-30% by weight of the total weight of active ingredients in the pharmaceutical composition.

[0183] 2. The amount of active ingredient in the immediate-release granules shall account for 72 to 77% by weight, preferably 75% by weight, of the total weight of the active ingredient in the pharmaceutical composition. The pharmaceutical composition according to item 1, wherein the amount of active ingredient in the sustained-release pellets accounts for 23 to 27% by weight, preferably 25% by weight, of the total weight of the active ingredient in the pharmaceutical composition.

[0184] 3. The content of the active ingredient in the immediate-release granules is 75-95% by weight, preferably 77-83% by weight, based on the weight of the immediate-release granules. The pharmaceutical composition according to any one of the preceding items, wherein the content of the active ingredient in the pellet core is 35 to 45% by weight, preferably 37 to 43% by weight, based on the weight of the pellet core.

[0185] 4. The pharmaceutical composition according to any one of the preceding items, wherein the subcoat content in the sustained-release pellet is 14 to 18% by weight, preferably 15% by weight, based on the weight of the pellet core.

[0186] 5. Sub-courts are based on the weight of the sub-court. i. 35-40% by weight, preferably 37.5% by weight of hydroxypropyl cellulose, ii. 35-40% by weight, preferably 37.5% by weight of hydroxypropyl methylcellulose, iii. A pharmaceutical composition according to any one of the preceding items, comprising 20-30% by weight, preferably 25% by weight, of talc.

[0187] 6. The sustained-release coating is measured by the weight of the sustained-release coating. 1) 97-100% by weight of a film-forming agent, 2) A pharmaceutical composition according to any one of the preceding items, comprising 0 to 3% by weight of a pore-forming material.

[0188] 7. The pharmaceutical composition according to any one of the preceding items, wherein the content of the sustained-release coating in the sustained-release pellet is 65 to 80% by weight, preferably 68 to 75% by weight, and more preferably 70% by weight, based on the weight of the pellet core provided with the subcoat.

[0189] 8. Immediate release granules are i. 90% by weight of valproic acid or valproate, ii. 9.5% by weight of anhydrous dicalcium phosphate, iii. Composed of 0.5% by weight of magnesium stearate, b) The pellet core is i. 40% by weight of valproic acid or valproate, ii. 59% by weight of microcrystalline cellulose, iii. Composed of 1% by weight of magnesium stearate, c) The sub-court is i. Provided in an amount equivalent to 15% of the weight of the pellet core, ii. The weight of the sub-court, 1) 37.5% by weight of hydroxypropyl cellulose, 2) 37.5% by weight of hydroxypropyl methylcellulose, 3) Containing 25% by weight of talc, d) The sustained-release coating is i. Provided in an amount equivalent to 70% of the weight of the pellet core coated with subcoat, ii. By weight of the sustained-release coating, 1) 98% ethylcellulose and 2) Composed of 2% pore-forming material containing hydroxypropyl methylcellulose, The amount of active ingredient in the immediate-release granules accounts for 75% by weight of the total weight of active ingredients in the pharmaceutical composition. The pharmaceutical composition according to any one of the preceding items, wherein the amount of active ingredient in the sustained-release pellets accounts for 25% by weight of the total weight of the active ingredient in the pharmaceutical composition.

[0190] 9. A pharmaceutical composition according to any one of the preceding items, further comprising a capsule for containing immediate-release granules and sustained-release pellets, wherein the capsule defines the unit volume of the pharmaceutical composition.

[0191] 10. A method for producing a pharmaceutical composition described in any one of the preceding items, i. Steps for producing immediate-release granules, ii. Steps for manufacturing the pellet core, iii. The pellet core, a. A substep of suspending or dissolving the subcoat in an aqueous solvent to form an aqueous solution or dispersion of the subcoat, b. A step of coating with a subcoat by performing a substep of bringing a pellet core into contact with an aqueous solution or dispersion of the subcoat to produce a pellet core with a subcoat, iv. A pellet core with a subcoat is provided. a. A substep of suspending or dissolving the sustained-release coating in an aqueous solvent to form an aqueous solution or dispersion of the sustained-release coating, A method comprising the steps of further coating a pellet core coated with a subcoat b with a sustained-release coating by performing a substep of contacting the pellet core coated with a subcoat b with an aqueous solution or dispersion of a sustained-release coating to produce a sustained-release pellet.

[0192] 11. A pharmaceutical composition used in a method of pretreatment of cancer, administered to a person with cancer, as described in any one of items 1 to 10.

[0193] 12. A pharmaceutical composition for use as described in item 11, wherein the cancer is selected from the group consisting of diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia, T-cell lymphoma, myeloma, and Hodgkin lymphoma.

[0194] 13. A pharmaceutical composition for use as described in item 11 or 12, which is administered to a human before treatment with chemotherapy and / or immunotherapy.

[0195] 14. A pharmaceutical composition for use as described in any one of items 11 to 13, administered twice daily.

[0196] 15. A pharmaceutical composition for use according to any one of items 11-14, wherein a steroid selected from the group consisting of prednisone, prednisolone, dexamethasone, and betamethasone is also administered to humans.

Claims

1. A pharmaceutical composition for oral administration, a) Immediate-release granules, i. An active ingredient selected from the group consisting of valproic acid, semisodium valproate, sodium valproate, and magnesium valproate, ii. Immediate-release granules containing a filler, b) A sustained-release pellet, i. A pellet core, (1) An active ingredient selected from the group consisting of valproic acid, semisodium valproate, sodium valproate, and magnesium valproate, (2) A pellet core containing a filler, ii. A subcoat provided on the pellet core, having a content of 10 to 20% by weight based on the weight of the pellet core, and containing a film-forming agent, iii. A sustained-release pellet comprising a sustained-release coating provided on the subcoat, the content of which is 25 to 100% by weight based on the weight of the pellet core coated with the subcoat, and which contains a film-forming agent, The amount of the active ingredient in the immediate-release granules accounts for 70 to 80% by weight of the total weight of the active ingredients in the pharmaceutical composition, and the amount of the active ingredient in the sustained-release pellets accounts for 20 to 30% by weight of the total weight of the active ingredients in the pharmaceutical composition. The film-forming agent of the sustained-release coating is a pharmaceutical composition that is different from the film-forming agent of the subcoat.

2. The amount of the active ingredient in the immediate-release granules is 72 to 77% by weight, preferably 75% by weight, of the total weight of the active ingredient in the pharmaceutical composition. The pharmaceutical composition according to claim 1, wherein the amount of the active ingredient in the sustained-release pellets accounts for 23 to 27% by weight, preferably 25% by weight, of the total weight of the active ingredient in the pharmaceutical composition.

3. The content of the active ingredient in the immediate-release granules is 75 to 95% by weight, preferably 77 to 83% by weight, based on the weight of the immediate-release granules. The pharmaceutical composition according to any one of the preceding claims, wherein the content of the active ingredient in the pellet core is 35 to 45% by weight, preferably 37 to 43% by weight, based on the weight of the pellet core.

4. The pharmaceutical composition according to any one of the preceding claims, wherein the subcoat content in the sustained-release pellet is 14 to 18% by weight, preferably 15% by weight, based on the weight of the pellet core.

5. The aforementioned subcoat is, by the weight of the aforementioned subcoat, i. 35-40% by weight, preferably 37.5% by weight of hydroxypropyl cellulose, ii. 35-40% by weight, preferably 37.5% by weight of hydroxypropyl methylcellulose, iii. A pharmaceutical composition according to any one of the preceding claims, comprising 20 to 30% by weight, preferably 25% by weight, of talc.

6. The sustained-release coating is, by weight of the sustained-release coating, 1) 97-100% by weight of a film-forming agent, 2) A pharmaceutical composition according to any one of the preceding claims, comprising 0 to 3% by weight of a pore-forming material.

7. The pharmaceutical composition according to any one of the preceding claims, wherein the content of the sustained-release coating in the sustained-release pellet is 65 to 80% by weight, preferably 68 to 75% by weight, and more preferably 70% by weight, based on the weight of the pellet core provided with the subcoat.

8. a) The immediate-release granules are, i. 90% by weight of valproic acid or valproate, ii. 9.5% by weight of anhydrous dicalcium phosphate, iii. Composed of 0.5% by weight of magnesium stearate, b) The pellet core is i. 40% by weight of valproic acid or valproate, ii. 59% by weight of microcrystalline cellulose, iii. Composed of 1% by weight of magnesium stearate, c) The subcoat is, i. Provided in an amount equivalent to 15% of the weight of the pellet core, ii. The weight of the aforementioned subcoat, 1) 37.5% by weight of hydroxypropyl cellulose, 2) 37.5% by weight of hydroxypropyl methylcellulose, 3) Contains 25% by weight of talc, d) The sustained-release coating is i. Provided in an amount equivalent to 70% of the weight of the pellet core coated with the subcoat, ii. By weight of the sustained-release coating, 1) 98% ethylcellulose and 2) Composed of 2% pore-forming material containing hydroxypropyl methylcellulose, The amount of the active ingredient in the immediate-release granules accounts for 75% by weight of the total weight of the active ingredient in the pharmaceutical composition. The pharmaceutical composition according to any one of the preceding claims, wherein the amount of the active ingredient in the sustained-release pellets accounts for 25% by weight of the total weight of the active ingredient in the pharmaceutical composition.

9. The pharmaceutical composition according to any one of the preceding claims, further comprising a capsule for containing the immediate-release granules and the sustained-release pellets, wherein the capsule defines a unit volume of the pharmaceutical composition.

10. A method for producing a pharmaceutical composition according to any one of the preceding claims, i. The step of producing the immediate-release granules, ii. The step of manufacturing the pellet core, iii. The pellet core, a. A substep of suspending or dissolving the subcoat in an aqueous solvent to form an aqueous solution or dispersion of the subcoat, b. A step of coating with the subcoat by performing a substep of bringing the pellet core into contact with an aqueous solution or dispersion of the subcoat to manufacture the pellet core having the subcoat provided, iv. The pellet core provided with the subcoat, a. A substep of suspending or dissolving the sustained-release coating in an aqueous solvent to form an aqueous solution or dispersion of the sustained-release coating, A method comprising the steps of: b) bringing the pellet core coated with the subcoat into contact with the aqueous solution or dispersion of the sustained-release coating to produce the sustained-release pellets; and b) further coating with the sustained-release coating.

11. The pharmaceutical composition according to any one of claims 1 to 10, used in a method of pretreatment for cancer, administered to a person suffering from cancer.

12. The pharmaceutical composition for use according to claim 11, wherein the cancer is selected from the group consisting of diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia, T-cell lymphoma, myeloma, and Hodgkin lymphoma.

13. The pharmaceutical composition for use according to claim 11 or 12, which is administered to a human before treatment with chemotherapy and / or immunotherapy.

14. The pharmaceutical composition for use according to any one of claims 11 to 13, wherein the pharmaceutical composition is administered twice a day.

15. A pharmaceutical composition for use according to any one of claims 11 to 14, wherein a steroid selected from the group consisting of prednisone, prednisolone, dexamethasone, and betamethasone is also administered to the human.

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