WO2025142803A1 - Pharmaceutical composition for oral administration of dnmt inhibitor - Google Patents

Abstract

The present disclosure provides a pharmaceutical composition for oral administration comprising core particles, a protective layer and an enteric layer, the pharmaceutical composition being characterized in that: (a) the core particles each comprise compound I and an additive; (b) the protective layer comprises an additive coated on the core particles; and (c) the enteric layer comprises an enteric polymer coated on the protective layer.

Description

Pharmaceutical compositions for oral administration of DNMT inhibitors

The present disclosure relates to an orally administered pharmaceutical composition that contains an orally administered DNMT inhibitor that has high stability against the hydrolytic metabolic enzyme cytidine deaminase and can replace 5-azacytidine or its 2'-deoxy derivative.

DNAMTs is an abbreviation for DNA-methyltransferases, a group of enzymes that catalyze the methylation of the amino group at position 6 of the adenine ring in DNA chains (Adenine N6-specific DNA-methyltransferase: EC 2.1.1.72), the methylation of the amino group at position 4 of the cytosine ring (Cytosine N4-specific DNA-methyltransferase: EC 2.1.1.113), or the methylation of the amino group at position 5 of the cytosine ring (Cytosine C5-specific DNA-methyltransferase: EC 2.1.1.37).
5-Azacytidine and its 2'-deoxy derivative (decitabine) are known as selective enzyme inhibitors (DNA methylation inhibitors, DNA demethylating agents) against DNA methyltransferase (DNMT).

Vidaza (registered trademark), a subcutaneous or intravenous formulation of 5-azacytidine indicated for the treatment of myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML), is usually administered to adults subcutaneously or by intravenous infusion over 10 minutes at 75 mg/ ^m2 (body surface area) of azacitidine once a day for seven days, followed by a three-week drug holiday, which constitutes one cycle, and this cycle is repeated.

Dacogen (registered trademark), an intravenous formulation of decitabine indicated for the treatment of myelodysplastic syndromes, is administered as a continuous intravenous infusion of 20 mg/ ^m2 of decitabine once a day over at least one hour for five consecutive days, followed by a 23-day rest period, in one cycle.

The above drugs are administered parenterally, either subcutaneously or intravenously, so patients who require treatment with the drugs must undergo multiple treatment cycles, which requires them to visit the hospital for long periods of time.

Clinical studies have been reported on administration methods that can maximize the demethylation effect while reducing side effects by reducing the dose of decitabine in one treatment cycle, shortening the intravenous administration time, and extending the administration interval (Non-Patent Documents 1 and 2). However, with the injectables used in these administration methods, it is still difficult to control the sudden rise in decitabine blood concentration or to maintain an effective blood concentration on a sustained basis.

In recent years, pharmaceutical compositions for oral administration have been discovered, such as oral combination drugs that combine decitabine with prodrugs that are metabolized to decitabine in the body after subcutaneous administration and have a pharmacological effect, or decitabine metabolic enzyme inhibitors that suppress the breakdown of decitabine when administered orally.

Guadecitabine (compound SGI-110) (Patent Documents 1-2), a prodrug of decitabine that is highly stable against the hydrolytic metabolic enzyme cytidine deaminase of decitabine, is highly polar due to its dinucleotide structure and is thought to have difficulty permeating cell membranes (Non-Patent Documents 3-4). Also, since its route of administration is subcutaneous, it is thought that there is still room for improvement as a pharmaceutical composition for oral administration.

It has been reported that the oral combination drug "INQOVI (registered trademark) (oral C-DEC, ASTX727)" (decitabine 35 mg/cedazuridine 100 mg) consisting of decitabine and the cytidine deaminase inhibitor cedazuridine, which is indicated for myelodysplastic syndrome and chronic myelomonocytic leukemia (CMML), shows pharmacokinetics, pharmacodynamics, safety and tolerability equivalent to that of decitabine administered intravenously at 20 mg/ ^m2 once a day for 5 days (Non-Patent Document 5). However, since the behavior of the blood concentration of this formulation is similar to that of Dacogen (registered trademark), an intravenous formulation, it is believed that there is still a need for a pharmaceutical composition for oral administration that can sustain the blood concentration.

U.S. Publication No. 2007072796 (Japanese Patent No. 5030958) International Publication No. 2013033176 (Japanese Patent No. 6038921 Specification

CANCER June 1, 2008/Volume112/Number 11 Blood (2007) 109(1):52-57 Oncotarget, 2017, Vol. 8, No. 2, pp. 2949-2959 Epigenetics, 2016, Vol. 11, No. 10, pp. 709-720 Blood, 2021, Vol. 138, No. 6, 3682-3685

The present disclosure aims to provide a pharmaceutical composition of a DNMT inhibitor that can solve the problems inherent in conventional tumor treatment methods using DNMT inhibitors, and that suppresses the decomposition of the compound under normal distribution and storage conditions for the formulation, since compound I represented by formula (I) is chemically unstable, and that is orally absorbable.

In order to solve the above problems, the present inventors prepared a composition (granules) in which core particles containing compound I represented by formula (I) and an additive are coated with a protective layer containing an additive, and this protective layer is coated with an enteric layer containing an enteric base. As a result, the inventors discovered for the first time that an oral pharmaceutical composition containing these granules exhibits good stability under storage conditions for distribution, and simultaneously achieves a favorable in vivo absorption rate and a reduced DNA methylation rate, thereby completing the present invention.

More surprisingly, they discovered that acid stability depends on the weight ratio of the additive and the main drug used in the core particles, and the thickness of the protective layer and the enteric layer, thus completing the present invention.

The present disclosure includes the following features.
[1]
An oral pharmaceutical composition having a core particle, a protective layer and an enteric layer, characterized in that (a) the core particle contains compound I represented by the following formula (I) and an additive, (b) the protective layer contains an additive coated on the core particle, and (c) the enteric layer contains an enteric polymer coated on the protective layer.
[2]
The oral pharmaceutical composition according to [1], wherein the additive of the core particle is at least one selected from the group consisting of excipients and binders.
[3]
The pharmaceutical composition for oral administration according to [2], wherein the excipient is at least one selected from the group consisting of lactose, D-mannitol, starch, partially pregelatinized starch, talc, low-substituted hydroxypropylcellulose, crystalline cellulose, and sucrose.
[4]

The pharmaceutical composition for oral administration according to [2], wherein the binder is at least one selected from the group consisting of polyethylene glycol, glyceryl monostearate, methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, povidone, sodium carmellose, pregelatinized starch, polyvinyl alcohol, and polyvinyl alcohol-acrylic acid-methacrylic acid copolymer.
[5]
The pharmaceutical composition for oral administration described in [1], wherein the additive of the protective layer is at least one selected from the group consisting of excipients and binders.
[6]
The pharmaceutical composition for oral administration according to [5], wherein the excipient is at least one selected from the group consisting of talc, titanium oxide, gelatin, light anhydrous silicic acid, hydrous silicon dioxide, and magnesium aluminometasilicate.
[7]
The pharmaceutical composition for oral administration according to [5], wherein the binder is at least one selected from the group consisting of methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, pregelatinized starch, povidone, sodium carmellose, polyvinyl alcohol, and polyvinyl alcohol-acrylic acid-methacrylic acid copolymer.
[8]
The oral pharmaceutical composition according to [1], wherein the enteric layer further contains an excipient and a plasticizer.
[9]
The oral pharmaceutical composition according to [1], wherein the enteric polymer is at least one selected from the group consisting of methacrylic acid copolymer LD, dry methacrylic acid copolymer LD, hypromellose acetate succinate, hypromellose phthalate, and carboxymethylethylcellulose.
[10]
The pharmaceutical composition for oral administration described in [8], wherein the excipient is selected from the group consisting of talc, titanium oxide, light anhydrous silicic acid, hydrous silicon dioxide, and magnesium aluminometasilicate.
[11]
The pharmaceutical composition for oral administration described in [8], wherein the plasticizer is at least one selected from the group consisting of triethyl citrate, glyceryl monostearate, triacetin, and polyethylene glycol.
[12]
The pharmaceutical composition for oral administration according to any one of [1] to [11], wherein the amount of compound I is 20 parts by mass or more per 100 parts by mass of the core particles.
[13]
The pharmaceutical composition for oral administration according to any one of [1] to [11], wherein the amount of the protective layer is 15 parts by mass or more per 100 parts by mass of the core particles.
[14]
The pharmaceutical composition for oral administration according to any one of [1] to [11], wherein the protective layer has a thickness of 15 μm or more.
[15]
The pharmaceutical composition for oral administration according to any one of [1] to [11], wherein the amount of the enteric layer is 14 parts by mass or more per 100 parts by mass of granules in which the core particle is coated with the protective layer.
[16]
The pharmaceutical composition for oral administration according to any one of [1] to [11], wherein the enteric layer has a thickness of 20 μm or more.
[17]
The pharmaceutical composition for oral administration according to any one of [1] to [11], wherein the amount of compound I is 20 parts by mass or more and the amount of the protective layer is 15 parts by mass or more relative to 100 parts by mass of the core particles, and the amount of the enteric layer is 14 parts by mass or more relative to 100 parts by mass of the granules in which the core particles are coated with the protective layer.
[18]
The pharmaceutical composition for oral administration according to any one of [1] to [11], wherein the amount of compound I is 20 parts by mass or more per 100 parts by mass of the core particles, the thickness of the protective layer is 15 μm or more, and the thickness of the enteric layer is 20 μm or more.
[19]
The pharmaceutical composition for oral administration according to any one of [1] to [18], wherein the composition having the core particle, the protective layer and the enteric layer is a granule.
[20]
The pharmaceutical composition for oral administration according to any one of [1] to [19], which contains the granules.
[21]
The pharmaceutical composition for oral administration according to any one of [1] to [20], which is a granule formulation.
[22]
The pharmaceutical composition for oral administration according to any one of [1] to [21], which is a capsule or stick formulation containing the granules or the granules.
[23]
The pharmaceutical composition for oral administration according to any one of [1] to [20], which is a tablet containing the granules.
[24]
The pharmaceutical composition for oral administration according to any one of [1] to [23], which is an agent for preventing or treating a tumor.
[25]
The pharmaceutical composition for oral administration according to any one of [1] to [23], which is an agent for preventing or treating myelodysplastic syndrome, chronic myelomonocytic leukemia, acute myelogenous leukemia, or chronic myelogenous leukemia.

According to the present disclosure, there is provided an oral pharmaceutical composition that is expected to reduce the DNA methylation rate while reducing the risk of side effects by being continuously absorbed into the body through oral administration in the treatment of tumors with DNMT inhibitors.
According to the present disclosure, in the treatment of tumors with DNMT inhibitors, an oral pharmaceutical composition is provided which is expected to provide significant convenience to patients who must repeatedly visit hospitals over a long period of time to receive treatment by intravenous administration.
According to the present disclosure, an oral pharmaceutical composition is provided that is expected to significantly improve the quality of life (QOL) of patients who need to undergo multiple treatment cycles in tumor treatment with a DNMT inhibitor by reducing the number of hospital visit cycles.

FIG. 1 shows the results of measuring the average plasma concentration of decitabine liberated from Compound I upon oral administration of Granule I of the present disclosure and the average plasma concentration of decitabine on the first day of intravenous administration (DAC in the figure is shown as an abbreviation for the generic name decitabine). FIG. 2 shows the results of LINE-1 methylation transition on the 5th day after oral administration of Granule I of the present disclosure and intravenous administration of decitabine (DAC in the figure is shown as an abbreviation for the generic name decitabine).

The oral pharmaceutical composition and the like of the present disclosure are described in detail below. However, the following description is merely an example for the purpose of explaining the present disclosure, and is not intended to limit the present disclosure to the scope of this description.

The present disclosure provides an oral pharmaceutical composition having a core particle, a protective layer, and an enteric layer, characterized in that (a) the core particle contains compound I represented by formula (I) and an additive, (b) the protective layer contains an additive coated on the core particle, and (c) the enteric layer contains an enteric polymer coated on the protective layer.

Core Particles The core particles of the present disclosure contain compound I represented by the following formula (I) and an additive.

Formula (I):

5'-O-triethylsilyl-2'-deoxy-5-azacytidine (also referred to as Compound I of the present disclosure, Compound I or OP-2100) represented by the formula (I) may be prepared, isolated or obtained by any method known to those skilled in the art. For example, it may be prepared according to the method described in Japanese Patent No. 6162349, the disclosure of which is incorporated herein by reference in its entirety. Compound I of the present disclosure is a prodrug that has high stability against the hydrolytic metabolic enzyme cytidine deaminase of decitabine and can slowly release the corresponding decitabine under physiological conditions.

Compound I of the present disclosure may be a crystal, and may have a single crystal form or a mixture of multiple crystal forms. Crystals of Compound I can be produced by crystallizing Compound I using a crystallization method known per se, a crystallization method disclosed in the present application, or a method equivalent thereto.

Compound I of the present disclosure may be a solvate (e.g., a hydrate, etc.), and both solvates and non-solvates (e.g., anhydrous, etc.) are included in Compound I. In some embodiments, Compound I of the present disclosure is a hemihydrate (also called a hemihydrate).

In some embodiments, the core particles of the present disclosure are disposed at the center of the oral pharmaceutical composition of the present disclosure, and the form of the core particles may be, but is not limited to, particles. In some embodiments, the average particle size of the core particles is preferably within the range of 50.0 to 950.0 μm, more preferably 400.0 to 900.0 μm, and even more preferably 700.0 to 850.0 μm.

The additives contained in the core particles of the present disclosure are not particularly limited, so long as they contain ingredients that are used as additives (inactive substances) in pharmaceuticals.

The additives contained in the core particles of the present disclosure may include at least one selected from the group consisting of excipients and binders.

In some embodiments, examples of excipients contained in the core particles of the present disclosure include, but are not limited to, lactose hydrate, anhydrous lactose, crystalline cellulose, D-mannitol, erythritol, xylitol, sorbitol, isomalt, maltitol, maltose, white sugar, sucrose, glucose, starch (corn starch, potato starch, rice starch, wheat starch, etc.), hydroxypropyl starch, pregelatinized starch, partially pregelatinized starch, talc, low-substituted hydroxypropyl cellulose, sodium carboxymethyl starch, dextrin, powdered reduced maltose syrup, ammonioalkyl methacrylate copolymer, ethyl cellulose, calcium hydrogen phosphate, etc.

In some embodiments, the excipient contained in the core particles of the present disclosure may include, but is not limited to, at least one selected from the group consisting of lactose, D-mannitol, starch, partially pregelatinized starch, talc, low-substituted hydroxypropyl cellulose, crystalline cellulose, and sucrose.

In some embodiments, examples of binders contained in the core particles of the present disclosure include, but are not limited to, alkyl celluloses (e.g., methyl cellulose), hydroxyalkyl celluloses (e.g., hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxybutyl cellulose), hydroxyalkyl alkyl celluloses (e.g., hydroxyethyl methyl cellulose, hypromellose), carboxyvinyl polymers, polyvinyl alcohol, polyvinyl alcohol-based copolymers (copolymers in which polyvinyl alcohol is one of the monomers, such as polyvinyl alcohol-acrylic acid-methyl methacrylate copolymers, polyvinyl alcohol-polyethylene glycol graft copolymers), polyvinylpyrrolidone, sodium carmellose, pregelatinized starch, and the like.

In some embodiments, examples of binders contained in the core particles of the present disclosure include, but are not limited to, at least one selected from the group consisting of polyethylene glycol, glyceryl monostearate, methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, povidone, sodium carmellose, pregelatinized starch, polyvinyl alcohol, and polyvinyl alcohol-acrylic acid-methacrylic acid copolymer.

In some embodiments, in the core particles of the present disclosure, the amount of compound I per 100 parts by weight of the core particles is, for example, 20 parts by weight or more, 30 parts by weight or more, or 40 parts by weight or more.

In some embodiments, in the core particle of the present disclosure, the amount of compound I is 20 parts by mass or more per 100 parts by mass of the core particle.
The upper limit of the amount of Compound I is preferably 60 parts by mass or less based on 100 parts by mass of the core particles.
In some embodiments, in the core particles of the present disclosure, the amount of Compound I is preferably 20 to 60 parts by mass, more preferably 25 to 55 parts by mass, relative to 100 parts by mass of the core particles.

Protective Layer The protective layer of the present disclosure is a suitable polymer intermediate layer coated on the surface of the core particle of the present disclosure (between the core particle and the enteric substrate) in order to suppress or prevent chemical decomposition of Compound I due to contact with the enteric substrate. The protective layer of the present disclosure is not particularly limited as long as it has the function of suppressing chemical decomposition of Compound I contained in the core particle of the present disclosure due to contact with the enteric substrate.

In some embodiments, in the protective layer of the present disclosure, the amount of protective layer per 100 parts by weight of core particles is, for example, 15 parts by weight or more, 20 parts by weight or more, or 25 parts by weight or more.

In some embodiments, in the protective layer of the present disclosure, the amount of the protective layer is 15 parts by mass or more per 100 parts by mass of the core particles.
The upper limit of the amount of the protective layer is preferably 45 parts by mass or less per 100 parts by mass of the core particles.
In some embodiments, in the protective layer of the present disclosure, the amount of the protective layer is preferably 15 to 45 parts by mass, and more preferably 15 to 40 parts by mass, per 100 parts by mass of the core particles.

In some embodiments, the protective layer of the present disclosure has a thickness of, for example, 15 μm or more, 20 μm or more, 25 μm or more, or 30 μm or more.
The upper limit of the thickness of the protective layer is preferably 50 μm or less.
In some embodiments, the thickness of the protective layer of the present disclosure is preferably 15 to 50 μm, and more preferably 20 to 40 μm.

In some embodiments, the thickness of the protective layer of the present disclosure may be, but is not limited to, about 32 μm.

In some embodiments, the protective layer of the present disclosure includes an additive coated on the core particles of the present disclosure.

In some embodiments, the protective layer of the present disclosure includes an additive coated directly onto the surface of the core particle of the present disclosure.

In some embodiments, the additive contained in the protective layer of the present disclosure is at least one selected from the group consisting of excipients and binders.

In some embodiments, the excipient contained in the protective layer of the present disclosure may include, but is not limited to, at least one selected from the group consisting of talc, titanium oxide, gelatin, light anhydrous silicic acid, hydrous silicon dioxide, and magnesium aluminometasilicate.

In some embodiments, the binder contained in the protective layer of the present disclosure may include, but is not limited to, at least one water-soluble polymer selected from the group consisting of methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, pregelatinized starch, povidone, sodium carmellose, polyvinyl alcohol, and polyvinyl alcohol-acrylic acid-methacrylic acid copolymer.

In some embodiments, the content of the excipient contained in the protective layer of the present disclosure is 60 parts by weight to 80 parts by weight per 100 parts by weight of the protective layer of the present disclosure.

The enteric layer of the present disclosure is a suitable polymer layer coated on the surface of the protective layer of the present disclosure in order to suppress or prevent the chemical decomposition of Compound I of the present disclosure from gastric acid. The enteric layer of the present disclosure is not particularly limited as long as it has the function of suppressing or preventing the chemical decomposition of Compound I contained in the core particle of the present disclosure from gastric acid.

In some embodiments, in the enteric layer of the present disclosure, the amount of the enteric layer is, for example, 14 parts by weight or more or 28 parts by weight or more per 100 parts by weight of granules in which a core particle is coated with a protective layer.

In some embodiments, in the enteric layer of the present disclosure, the amount of the enteric layer is 14 parts by mass or more per 100 parts by mass of the granules in which the core particles are coated with the protective layer.
The upper limit of the amount of the enteric layer is preferably 45 parts by weight or less per 100 parts by weight of granules in which the core particles are coated with a protective layer.
In some embodiments, in the enteric layer of the present disclosure, the amount of the enteric layer is preferably 14 to 45 parts by weight, more preferably 14 to 40 parts by weight, per 100 parts by weight of the granules in which the core particles are coated with the protective layer.

In some embodiments, the enteric layer of the present disclosure has a thickness of, for example, 20 μm or more, 30 μm or more, or 40 μm or more.
The upper limit of the thickness of the enteric layer is preferably 50 μm or less.
In some embodiments, the enteric layer of the present disclosure has a thickness preferably between 20 and 50 μm, more preferably between 20 and 40 μm.

In some embodiments, the enteric layer of the present disclosure has a thickness of, but is not limited to, about 38 μm.

The enteric layer of the present disclosure is a polymer layer containing an enteric polymer that is coated on the protective layer of the present disclosure.

In some embodiments, the enteric polymer contained in the enteric layer of the present disclosure may include, but is not limited to, at least one selected from the group consisting of methacrylic acid copolymer LD, dry methacrylic acid copolymer LD, hypromellose acetate succinate, hypromellose phthalate, and carboxymethylethylcellulose.

In some embodiments, the enteric layer of the present disclosure may further include excipients and plasticizers.

In some embodiments, the excipient contained in the enteric layer of the present disclosure may include, but is not limited to, at least one selected from the group consisting of talc, titanium oxide, light anhydrous silicic acid, hydrous silicon dioxide, and magnesium aluminometasilicate.

In some embodiments, the plasticizer contained in the enteric layer of the present disclosure may include, but is not limited to, at least one selected from the group consisting of triethyl citrate, glyceryl monostearate, triacetin, and polyethylene glycol.

In the present disclosure, the amount of the additives of the present disclosure is not particularly limited as long as it does not impair the effect of the oral pharmaceutical composition of the present disclosure.

The present disclosure provides a tumor prevention/treatment agent that includes the oral pharmaceutical composition of the present disclosure.

In some embodiments, the present disclosure provides a preventive or therapeutic agent for myelodysplastic syndrome, chronic myelomonocytic leukemia, acute myeloid leukemia, or chronic myeloid leukemia, comprising an oral pharmaceutical composition of the present disclosure.

The composition comprised of the core particles, protective layer and enteric layer of the present disclosure forms granules.
The pharmaceutical composition of the present disclosure is an oral pharmaceutical composition containing the granules.
The dosage form of the oral pharmaceutical composition of the present disclosure is not particularly limited as long as it contains the granules, and examples thereof include, but are not limited to, granules containing the granules, capsules encapsulating the granules, tablets encompassing the granules, or sticks encapsulating the granules.

In some embodiments, the present disclosure provides granules or capsules containing granules.

In some embodiments, the present disclosure provides a tablet containing granules and an additive.

In some embodiments, the additives contained in the tablets of the present disclosure may include commonly used stabilizers, lubricants, binders, excipients, disintegrants, etc., and other additives such as flavoring agents and colorants may also be used as needed.

In some embodiments, examples of stabilizers contained in the tablets of the present disclosure include, but are not limited to, hydrous silicon dioxide, citric acid hydrate, light anhydrous silicic acid, macrogol 4000, etc.

In some embodiments, lubricants contained in the tablets of the present disclosure may include, but are not limited to, hydrogenated oils, calcium stearate, magnesium stearate, talc, sodium stearyl fumarate, etc.

In some embodiments, binders contained in the tablets of the present disclosure may include, but are not limited to, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose, povidone, etc.

In some embodiments, excipients contained in the tablets of the present disclosure may include, but are not limited to, crystalline cellulose, lactose, crystalline cellulose, corn starch, potato starch, D-mannitol, white sugar, sucrose, glucose, etc.

In some embodiments, disintegrants contained in the tablets of the present disclosure include, but are not limited to, carboxymethylcellulose, calcium carboxymethylcellulose, sodium carboxymethylstarch, crospovidone, low-substituted hydroxypropylcellulose, croscarmellose sodium, partially pregelatinized starch, etc.

In some embodiments, flavoring agents contained in the tablets of the present disclosure include, but are not limited to, sucralose, aspartame, etc.

In some embodiments, colorants contained in the tablets of the present disclosure may include, but are not limited to, ferric oxide, yellow ferric oxide, etc.

The oral pharmaceutical composition disclosed herein can be prepared by common manufacturing methods in the art depending on the dosage form.

The oral pharmaceutical composition of the present disclosure can be prepared, for example, by the following steps.

(Step of Producing Core Particles)
In some embodiments, the core particles of the present disclosure can be produced by putting additives (e.g., excipients, etc.) into a fluidized bed granulator, and coating the suspension of compound I of the present disclosure and additives (e.g., binders, etc.) while spraying and drying.

(Step of coating the core particles with a protective layer)
In some embodiments, the protective layer of the present disclosure can be produced by coating the granules obtained above (core particles of the present disclosure) with a suspension of additives (e.g., excipients, binders, etc.) while spraying and drying.

(Step of coating the protective layer with an enteric layer)
In some embodiments, the enteric layer of the present disclosure can be prepared by coating the above obtained granules (the protective layer of the present disclosure) with a suspension containing additives (e.g., enteric polymers, excipients, plasticizers, etc.) while spraying and drying.

Granules of the present disclosure
The granules of the present disclosure are produced through a process of producing the above-mentioned core particles, a process of coating the periphery of the core particles with a protective layer, and a process of coating the periphery of the protective layer with an enteric layer.
The granules of the present disclosure can be used as an oral pharmaceutical composition by themselves.Also, the granules of the present disclosure can be used to manufacture oral pharmaceutical compositions having dosage forms such as granules, tablets, capsules, sticks, etc., using a general method in the technical field of pharmaceutical preparations.

The tablets of the present disclosure can be manufactured using the granules of the present disclosure by a common tablet manufacturing method in the art.

In some embodiments, the granules of the present disclosure can be mixed with additives (e.g., stabilizers, lubricants, binders, excipients, disintegrants, flavorings, colorants, etc.) and then compressed using a tablet press. The compression pressure when compressing using a tablet press is preferably 1 kN to 10 kN, and more preferably 1 kN to 5 kN.

The capsule formulation of the present disclosure can be produced by filling the granules of the present disclosure into capsules using a filling method commonly used in the art.

In some embodiments, the capsule formulation of the present disclosure can be prepared by filling the granules of the present disclosure into capsules using common filling methods in the art.

The capsules of the present disclosure may be conventional capsules used in the art. In some embodiments, the capsules of the present disclosure may be, but are not limited to, gelatin capsules, hydroxypropylmethylcellulose (HPMC) capsules, pullulan capsules, etc. (e.g., hydroxypropylmethylcellulose (HPMC) capsules), Licaps™ capsules, Vcaps™ capsules, Coni-Snap™ capsules, Press-fit™ capsules, and Xpress-fit™ capsules.

The capsule formulation of the present disclosure may be, for example, but not limited to, a capsule size of No. 1, a capsule size of No. 2, a capsule size of No. 3, a capsule size of No. 4, and a capsule size of No. 5. In some embodiments, a capsule size of No. 4 or a capsule size of No. 5 is selected from the viewpoint of ease of administration.

The stick formulation of the present disclosure can be produced by filling the granules of the present disclosure into a stick package as is common in the art.

The present disclosure will be explained below using examples, but the present disclosure is not limited to the following examples.

199.2 g of crystalline cellulose granules were placed in a fluidized bed granulator, and a solution of 40.0 g of methylcellulose dissolved in 1,240 g of purified water was dispersed and suspended in 182.4 g of compound I (as 1/2 hydrate), which was then sprayed and dried to obtain granule A. Next, a solution of 22.0 g of hydroxypropylmethylcellulose dissolved in 550 g of purified water was dispersed and suspended in 88.2 g of talc, which was then sprayed and dried to obtain granule B. Finally, a solution of 12.88 g of glycerin monostearate, 6.48 g of triethyl citrate, and 1.3 g of polysorbate 80 dissolved in 300 g of purified water was added with 430.7 g of methacrylic acid copolymer LD (solid content: 129.2 g), which was gently mixed and stirred, and then sprayed and dried to obtain granule I.

220.5 g of crystalline cellulose granules were placed in a fluidized bed granulator, and a solution of 25.0 g of methylcellulose dissolved in 775 g of purified water, into which 114.0 g of compound I (as 1/2 hydrate) was dispersed and suspended, was sprayed and dried to obtain granules A. Next, a solution of 19.9 g of hydroxypropylmethylcellulose dissolved in 497 g of purified water, into which 79.7 g of talc was dispersed and suspended, was sprayed and dried to obtain granules B. Finally, a solution of 11.85 g of glycerin monostearate, 5.9 g of triethyl citrate, and 1.2 g of polysorbate 80 dissolved in 273 g of purified water was added 394.3 g of methacrylic acid copolymer LD (solid content: 118.3 g), which was gently mixed and stirred, was sprayed and dried to obtain granules II.

240.0 g of crystalline cellulose granules were placed in a fluidized bed granulator, and a solution of 20.0 g of methylcellulose dissolved in 620 g of purified water and 91.2 g of compound I (as 1/2 hydrate) dispersed and suspended in the solution was sprayed and dried to obtain granule A. Next, a solution of 20.32 g of hydroxypropylmethylcellulose dissolved in 507 g of purified water and 81.24 g of talc dispersed and suspended in the solution was sprayed and dried to obtain granule B. Finally, a solution of 12.12 g of glycerin monostearate, 6.04 g of triethyl citrate, and 1.2 g of polysorbate 80 dissolved in 279 g of purified water was added with 403.3 g of methacrylic acid copolymer LD (solid content: 121.0 g), which was gently mixed and stirred, and the solution was sprayed and dried to obtain granule III.

204.0 g of crystalline cellulose granules were placed in a fluidized bed granulator, and 77.52 g of compound I (as 1/2 hydrate) was dispersed and suspended in a solution of 17.0 g of methylcellulose dissolved in 527 g of purified water, which was then sprayed and dried to obtain granule A. Next, 42.16 g of talc was dispersed and suspended in a solution of 10.54 g of hydroxypropylmethylcellulose dissolved in 263.5 g of purified water, which was then sprayed and dried to obtain granule B. Finally, 326.4 g of methacrylic acid copolymer LD (solid content: 97.92 g) was added to a solution of 9.79 g of glycerin monostearate, 4.87 g of triethyl citrate, and 0.98 g of polysorbate 80 dissolved in 225.8 g of purified water, and the mixture was gently mixed and stirred, and then sprayed and dried to obtain granule IV.

199.2 g of crystalline cellulose granules were placed in a fluidized bed granulator, and a solution of 40.0 g of methylcellulose dissolved in 1240 g of purified water was dispersed and suspended in 182.4 g of compound I (as 1/2 hydrate), which was then sprayed and dried to obtain granule A. Next, a solution of 22.0 g of hydroxypropylmethylcellulose dissolved in 550 g of purified water was dispersed and suspended in 88.2 g of talc, which was then sprayed and dried to obtain granule B. Finally, a solution of 6.4 g of glycerin monostearate, 3.2 g of triethyl citrate, and 0.64 g of polysorbate 80 dissolved in 148.1 g of purified water was added with 212.27 g of methacrylic acid copolymer LD (solid content: 63.68 g), which was gently mixed and stirred, and the solution was sprayed and dried to obtain granule V.

200.8 g of crystalline cellulose granules were placed in a fluidized bed granulator, and a solution of 60.0 g of methylcellulose dissolved in 1,860 g of purified water was dispersed and suspended in 205.3 g of compound I (as 1/2 hydrate), which was then sprayed and dried to obtain granule A. Next, a solution of 42.0 g of hydroxypropylmethylcellulose dissolved in 630 g of purified water was dispersed and suspended in 78.0 g of talc, which was then sprayed and dried to obtain granule B. Finally, a solution of 14.0 g of glycerin monostearate, 8.0 g of triethyl citrate, and 1.0 g of polysorbate 80 dissolved in 440 g of purified water was added with 470.0 g of methacrylic acid copolymer LD (solid content: 141.0 g), which was gently mixed and stirred, and the solution was sprayed and dried to obtain granule VI.

41.7 g of Granule VI (Example 6), 181.8 g of lactose hydrate, 75.0 g of crystalline cellulose, and 1.5 g of magnesium stearate were mixed in a polyethylene bag, and then compressed in a tablet press at a compression pressure of 3 kN to obtain tablets (circular tablets, diameter 12.0 mm, thickness 5.0 mm) each weighing 600 mg.

Compound I (as a hemihydrate) can be prepared, for example, by the following method.
A white solid of compound (I) was obtained according to the method described in Japanese Patent No. 6162349. 1.71 g (5 mmol) of the white solid of compound (I) was dissolved in 15 mL of acetone by heating, and then, while heating, 180 μL (10 mmol) of purified water and 75 mL of MTBE (methyl tert-butyl ether) were added and stirred overnight at room temperature. The resulting white solid was filtered to obtain 1.07 g (63% recovery) of crystalline powder (fine columnar crystals) of compound I (1/2 hydrate). The HPLC purity of the obtained crystalline powder was >99%. The crystalline powder exhibited a powder X-ray diffraction pattern with characteristic diffraction peaks at angles 2θ=5.6°±0.2°, 12.5°±0.2°, 12.9°±0.2°, 13.2°±0.2°, 15.0°±0.2°, 15.4°±0.2°, 15.8°±0.2°, 16.1°±0.2°, 17.1°±0.2°, 19.1°±0.2°, 20.4°±0.2°, 22.4°±0.2°, 23.4°±0.2°, 25.1°±0.2°, 25.9°±0.2°, and 28.1°±0.2° with CuKα radiation of wavelength 1.5419 Å.

Comparative Example 1

438.5 g of crystalline cellulose granules were placed in a fluidized bed granulator, and a solution of 5.0 g of methylcellulose dissolved in 155 g of purified water, into which 22.8 g of compound I (as 1/2 hydrate) was dispersed and suspended, was sprayed and dried to obtain granule A. Next, a solution of 30.0 g of hydroxypropylmethylcellulose dissolved in 750 g of purified water, into which 120.0 g of talc was dispersed and suspended, was sprayed and dried to obtain granule B. Finally, a solution of 19.2 g of glycerin monostearate, 9.6 g of triethyl citrate, and 1.92 g of polysorbate 80 dissolved in 450 g of purified water was added with 640.0 g of methacrylic acid copolymer LD (solid content: 192.0 g), which was gently mixed and stirred, and the solution was sprayed and dried to obtain granule VII.

Comparative Example 2

204.0 g of crystalline cellulose granules were placed in a fluidized bed granulator, and a solution of 17.0 g of methylcellulose dissolved in 527 g of purified water and 77.52 g of compound I (as 1/2 hydrate) dispersed and suspended in the solution was sprayed and dried to obtain granule A. Next, a solution of 4.08 g of hydroxypropylmethylcellulose dissolved in 102 g of purified water and 16.32 g of talc dispersed and suspended in the solution was sprayed and dried to obtain granule B. Finally, a solution of 9.18 g of glycerin monostearate, 4.59 g of triethyl citrate, and 0.92 g of polysorbate 80 dissolved in 211.5 g of purified water was added with 306.0 g of methacrylic acid copolymer LD (solid content: 91.80 g), which was gently mixed and stirred, and the solution was sprayed and dried to obtain granule VIII.

Comparative Example 3

438.5 g of crystalline cellulose granules were placed in a fluidized bed granulator, and a solution of 5.0 g of methylcellulose dissolved in 155 g of purified water, into which 22.8 g of compound I (as 1/2 hydrate) was dispersed and suspended, was sprayed and dried to obtain granule A. Next, a solution of 7.5 g of hydroxypropylmethylcellulose dissolved in 187.5 g of purified water, into which 30.0 g of talc was dispersed and suspended, was sprayed and dried to obtain granule B. Finally, a solution of 16.8 g of glycerin monostearate, 8.4 g of triethyl citrate, and 1.68 g of polysorbate 80 dissolved in 390 g of purified water was added 560.0 g of methacrylic acid copolymer LD (solid content: 168.0 g), which was gently mixed and stirred, was sprayed and dried to obtain granule IX.

The formulations of Examples 1 to 6 and Comparative Examples 1 to 3 are shown in Table 1.
Table 1

Test Example 1

Granule I (Example 1) was orally administered once a day for five days to male cynomolgus monkeys at four different doses (2.9 mg/kg, 5.8 mg/kg, 11.6 mg/kg, 23.2 mg/kg). 1 mL of blood was collected at 30, 60, 90, 120, 150, 180, 240, 300, 360, and 420 minutes on the first day of administration, and the average plasma concentrations of Compound I and released decitabine were measured.

For comparison, a study was conducted in which decitabine was administered intravenously (infusion) once daily for 1 hour to male cynomolgus monkeys at two doses (0.84 mg/kg and 1.67 mg/kg) for 5 consecutive days, and 1 mL of blood was collected at 30, 60, 90, 120, 150, 180, 240, 300, 360, and 420 minutes on the first day of administration to measure the average plasma concentration of decitabine. The dose of 0.84 mg/kg of decitabine was prepared by diluting a 16.7 mg/mL DMSO solution of decitabine 100-fold with a 1% cyclodextrin solution and administering it according to the body weight of the cynomolgus monkey. A dose of 1.67 mg/kg of decitabine was prepared by diluting a 33.4 mg/mL DMSO solution of decitabine 100-fold with a 1% cyclodextrin solution, and administered according to the body weight of the cynomolgus monkey.

The results of measuring the average plasma concentration of decitabine released from Compound I on the first day of oral administration of Granule I and the average plasma concentration on the first day of 1-hour intravenous administration of decitabine are shown in Figure 1. The average plasma concentration of decitabine released from Compound I when Granule I was administered orally showed a sustained behavior compared to when decitabine was administered intravenously, and the AUC and Cmax increased dose-dependently.

Test Example 2

In all treatment groups in Test Example 1, 2 mL of blood was collected prior to treatment and on days 1, 5, 8, 10, 12, 15, 19, 22, 26, and 29 of treatment. DNA was extracted from 200 μL of collected whole blood using a QIAamp DNA mini Kit (QIAGEN). The extracted DNA was processed according to the protocol for the EZ DNA Methylation Kit (ZYMO RESEARCH). PCR was performed on this sample according to the protocol of the Monkey Line-1 Methylation Assay Kit (EPIGEN Dx), and the CpG island cytosine methylation rate of LINE-1 (Long interspersed nucleotide factor-1) was measured by pyrosequencing. The methylation rates of four CpG sites were measured, and the average value was calculated as the methylation rate. The difference from the value before administration in each administration group, which was set to 0, was defined as the LINE-1 CpG demethylation.

The results of the transition in LINE-1 methylation on the fifth day after oral administration of Granule I and intravenous administration of decitabine are shown in Figure 2. The LINE-1 methylation level was observed over time after both oral administration of Granule I and intravenous administration of decitabine. The LINE-1 methylation level reached a maximum around 10 days after administration. Thereafter, the LINE-1 methylation level recovered to the same level as before administration. Furthermore, with oral administration of Granule I, the degree of methylation increased with increasing dose.

Test Example 3

Granule I (Example 1), Granule II (Example 2), Granule III (Example 3), Granule IV (Example 4), Granule V (Example 5), Granule VI (Example 6), Granule VII (Comparative Example 1), Granule VIII (Comparative Example 2) and Granule IX (Comparative Example 3) were filled into transparent glass bottles, then placed in aluminum bags and heat-sealed. After storing the bottles sealed at 50°C for 14 days, a purity test was performed. The total amount (total amount of related substances) (decomposition products of compound I) and the amount of decitabine produced in each granule were measured at the start of the test, and at 3, 7 and 14 days after the start of the test. The results are shown in Table 2.

The sample solution for the measurement of related substances was prepared by taking an amount corresponding to about 22.8 mg of Compound I of the present disclosure, adding 20 mL of N,N-dimethylformamide, and subjecting the solution to ultrasonic treatment for 10 minutes. The solution was then filtered through a membrane filter with a pore size of 0.45 μm or less, and the first 5 mL of the filtrate was removed to obtain the next filtrate. 10 μL of this solution was subjected to a test by liquid chromatography under the following conditions. The peak areas of each of the sample solutions were measured by the automatic integration method, and the total amount (total related amount) and the amount of decitabine were calculated by the area percentage method.
Test conditions: HPLC method (detector: ultraviolet spectrophotometer (measurement wavelength: 254 nm))
Table 2

Granules I, II, III, IV, V, and VI had significantly less increase in related substances (decomposition products of compound I) and suppressed the increase in related substances (decomposition products of compound I) compared to granule VII, which has a smaller amount of compound I relative to the amount of core particles, granule VIII, which has a thinner protective layer, and granule IX, which has a smaller amount of compound I relative to the amount of core particles and a thinner protective layer.

Test Example 4

An amount corresponding to about 22.8 mg of Compound I of the present disclosure was taken, 20 mL of N,N-dimethylformamide was added, and the mixture was sonicated for 10 minutes. The liquid was then filtered through a membrane filter with a pore size of 0.45 μm or less, the first 5 mL of the filtrate was removed, and the next filtrate was used as a sample solution. 10 μL of this liquid was subjected to a liquid chromatography test under the following conditions. Each peak area of the sample solution was measured by an automatic integration method, and the total amount (total related amount) and the amount of decitabine were calculated by the area percentage method.
Test conditions: HPLC method (detector: ultraviolet spectrophotometer (measurement wavelength: 254 nm))

Granules VI (Example 6) in an amount corresponding to about 41.05 mg of Compound I were filled into No. 4 hypromellose capsules, placed in polyethylene containers, and sealed with polypropylene caps equipped with desiccant. Purity tests were then carried out after storage at 40° C. and 75% RH for 6 months and at 25° C. and 60% RH for 12 months. The results of the purity tests, which measured the total amount (total amount of related substances) (decomposition products of Compound I) and the amount of decitabine produced in the granules at each time point, are shown in Table 3.
Table 3

Even in the commercial formulation of Granule VI, the increase in related substances (decomposition products of Compound I) was extremely small, demonstrating that the drug is stable even under market distribution.

Test Example 5

The dissolution rate of Compound I 120 minutes after the start of the test was determined for Granule VI (Example 6) and Tablet (Example 7) in an amount corresponding to approximately 22.8 mg of Compound I using the dissolution test method (paddle method) of the Japanese Pharmacopoeia, 18th Edition, General Test Methods, and the results (n=3) are shown in Table 4.

Test conditions: GC method (detector: FID)
Test solution: Japanese Pharmacopoeia No. 1 solution (dissolution test No. 1 solution, pH 1.2)
Test liquid volume: 900 mL
Paddle rotation speed: 50 rpm
Liquid temperature: 37°C
Table 4

The dissolution rate of compound I in the tablet in an acidic environment was suppressed to a low level similar to that of granules VI, demonstrating that the formulation containing the granules of the present disclosure has the function of suppressing or preventing the chemical decomposition of compound I in gastric acid even in tablet form.

Claims (25)

An oral pharmaceutical composition having a core particle, a protective layer and an enteric layer, characterized in that (a) the core particle contains compound I represented by formula (I) and an additive, (b) the protective layer contains an additive coated on the core particle, and (c) the enteric layer contains an enteric polymer coated on the protective layer.
The oral pharmaceutical composition according to claim 1, wherein the additive for the core particles is at least one selected from the group consisting of excipients and binders. The oral pharmaceutical composition according to claim 2, wherein the excipient is at least one selected from the group consisting of lactose, D-mannitol, starch, partially pregelatinized starch, talc, low-substituted hydroxypropyl cellulose, crystalline cellulose, and sucrose. The pharmaceutical composition for oral administration according to claim 2, wherein the binder is at least one selected from the group consisting of polyethylene glycol, glyceryl monostearate, methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, povidone, carmellose sodium, pregelatinized starch, polyvinyl alcohol, and polyvinyl alcohol-acrylic acid-methacrylic acid copolymer. The oral pharmaceutical composition according to claim 1, wherein the additive in the protective layer is at least one selected from the group consisting of excipients and binders. The oral pharmaceutical composition according to claim 5, wherein the excipient is at least one selected from the group consisting of talc, titanium oxide, gelatin, light anhydrous silicic acid, hydrous silicon dioxide, and magnesium aluminometasilicate. The pharmaceutical composition for oral administration according to claim 5, wherein the binder is at least one selected from the group consisting of methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, pregelatinized starch, povidone, carmellose sodium, polyvinyl alcohol, and polyvinyl alcohol-acrylic acid-methacrylic acid copolymer. The oral pharmaceutical composition of claim 1, wherein the enteric layer further comprises an excipient and a plasticizer. The oral pharmaceutical composition according to claim 1, wherein the enteric polymer is at least one selected from the group consisting of methacrylic acid copolymer LD, dry methacrylic acid copolymer LD, hypromellose acetate succinate, hypromellose phthalate, and carboxymethylethylcellulose. The oral pharmaceutical composition according to claim 8, wherein the excipient is selected from the group consisting of talc, titanium oxide, light anhydrous silicic acid, hydrous silicon dioxide, and magnesium aluminometasilicate. The oral pharmaceutical composition according to claim 8, wherein the plasticizer is at least one selected from the group consisting of triethyl citrate, glyceryl monostearate, triacetin, and polyethylene glycol. The oral pharmaceutical composition according to any one of claims 1 to 11, wherein the amount of compound I is 20 parts by mass or more per 100 parts by mass of the core particles. The oral pharmaceutical composition according to any one of claims 1 to 11, wherein the amount of the protective layer is 15 parts by mass or more per 100 parts by mass of the core particles. The oral pharmaceutical composition according to any one of claims 1 to 11, wherein the protective layer has a thickness of 15 μm or more. The oral pharmaceutical composition according to any one of claims 1 to 11, wherein the amount of the enteric layer is 14 parts by mass or more per 100 parts by mass of granules in which the core particles are coated with a protective layer. The oral pharmaceutical composition according to any one of claims 1 to 11, wherein the enteric layer has a thickness of 20 μm or more. The oral pharmaceutical composition according to any one of claims 1 to 11, wherein the amount of compound I is 20 parts by mass or more per 100 parts by mass of core particles, the amount of the protective layer is 15 parts by mass or more, and the amount of the enteric layer is 14 parts by mass or more per 100 parts by mass of granules in which the core particles are coated with the protective layer. The oral pharmaceutical composition according to any one of claims 1 to 11, wherein the amount of compound I is 20 parts by mass or more per 100 parts by mass of the core particles, the thickness of the protective layer is 15 μm or more, and the thickness of the enteric layer is 20 μm or more. The oral pharmaceutical composition according to any one of claims 1 to 18, wherein the composition having the core particle, the protective layer and the enteric layer is a granule. The oral pharmaceutical composition according to any one of claims 1 to 19, which contains the granules. The oral pharmaceutical composition according to any one of claims 1 to 20, which is a granule. The oral pharmaceutical composition according to any one of claims 1 to 21, which is a capsule or stick formulation containing the granules or granules. The oral pharmaceutical composition according to any one of claims 1 to 20, which is a tablet containing the granules. The oral pharmaceutical composition according to any one of claims 1 to 23, which is a tumor preventive or therapeutic agent. The oral pharmaceutical composition according to any one of claims 1 to 23, which is a preventive or therapeutic agent for myelodysplastic syndrome, chronic myelomonocytic leukemia, acute myeloid leukemia, or chronic myeloid leukemia.