A mercaptopurine microtablet, its preparation method and application

Abstract

The present invention discloses a mercaptopurine microtablet, its preparation method, and application. The mercaptopurine microtablet has a diameter of 2 to 3 mm and a diameter-to-thickness ratio of 0.85 to 1.15. It comprises the following components, calculated by weight: 15 to 30 parts of a mercaptopurine API; 40 to 80 parts of a filler; 3 to 8 parts of a binder; 1 to 3 parts of a lubricant; and 1 to 3 parts of a disintegrant. The D90 particle size of the mercaptopurine API is 40 to 125 μm; and the D50 particle size of the mercaptopurine API is 5 to 35 μm. The mercaptopurine microtablet of the present invention has excellent in vivo pharmacokinetic properties, good dissolution effect, excellent friability, hardness, and content uniformity. It is bioequivalent to the control tablet Purinethol in terms of absorption rate and extent, and has good application prospects in tumor treatment, especially in childhood acute lymphoblastic leukemia.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, in particular to a mercaptopurine micro-tablet and a preparation method and application thereof. Background Art

[0002] Mercaptopurine (6-purinethiol, 6-mercaptopurine, 6-Mercaptopurine, 6-MP) is a yellow crystalline powder. It is a cell cycle-specific drug that inhibits the purine biosynthesis pathway. Its chemical structure is similar to that of hypoxanthine, enabling it to competitively inhibit the conversion of hypoxanthine. Upon entry into the body, mercaptopurine must be converted to 6-mercaptopurine ribonucleotide by phosphoribosyltransferase within cells to become active. Mercaptopurine inhibits purine nucleotide synthesis and metabolism by inhibiting phosphoribosylpyrophosphate amidotransferase (PRPP amidotransferase), the rate-limiting enzyme in purine biosynthesis. Mercaptopurine can alter the synthesis and function of RNA and DNA and is a widely used anti-tumor agent.

[0003] Prior art reports show that mercaptopurine acts on a variety of malignant tumor cells by reducing the expression level of GLT8D1 protein, thereby reducing the level of PD-L1 protein, thereby relieving tumor immunosuppression and inhibiting the occurrence and development of tumors. Specifically, in the experiment, when different malignant tumor cell lines were treated with mercaptopurine, such as lung cancer cell lines H1299 and A549, colorectal cancer cell lines HCT116, DLD-1 and Lovo, liver cancer cell line Huh-7, breast cancer cell lines MDAMB-231 and SUM149 or MDA-MB-453, and melanoma cell lines A375 and SK-MEL-5, as the mercaptopurine concentration increased, the expression of GLT8D1 protein decreased, and at the same time, the protein level and protein molecular weight of PD-L1 in the cells changed significantly. The total amount of PD-L1 protein was significantly reduced, the molecular weight of PD-L1 protein became smaller due to the reduced glycosylation level, and the expression of PD-L1 on the cell membrane surface was also significantly reduced. When tumor cells were treated with mercaptopurine and co-incubated with activated PBMCs (peripheral blood mononuclear cells), the killing effect of PBMCs on tumor cells was significantly increased.

[0004] At present, mercaptopurine is mainly used in clinical practice for choriocarcinoma, malignant hydatidiform mole, acute lymphocytic leukemia (ALL), acute non-lymphocytic leukemia, and the blast crisis of chronic myeloid leukemia. Among them, leukemia is the malignant tumor with the highest incidence in children. Children aged 1-4 years old account for nearly 30% of all confirmed cases, and ALL accounts for more than 70% of childhood leukemia.

[0005] Mercaptopurine tablets are a backbone drug for the treatment of childhood acute lymphoblastic leukemia, but their clinical application faces numerous challenges. The only tablet strength available on the domestic market is 50mg, which cannot meet the personalized dosing requirements for children. Children must manually split the 50mg tablets, a process that can alter the drug's physical and chemical properties and increase the risk of contamination. For children, there is also the risk of inaccurate dispensing and administration, raising concerns about safety and efficacy. For healthcare professionals and patients' families, the cytotoxic nature of this drug poses a risk of drug exposure.

[0006] The prior art has disclosed a mercaptopurine microtablet and its preparation process, but the preparation process is cumbersome and complicated, and the use of organic solvents poses the risk of deflagration and explosion. At the same time, due to the lack of strict control over the specifications of the microtablets, it still cannot meet the requirements of personalized dosing for children and there are problems with in vivo absorption. In addition, the prepared microtablets also have poor dissolution effect and cannot take into account the hardness, friability and content uniformity of the microtablets, which has a great impact on the quality and efficacy of the drug.

[0007] At the same time, microtablets cannot be simply regarded as drugs that are proportionally scaled down from ordinary tablets, because the precision of ordinary tablet preparation equipment cannot meet the preparation requirements of microtablets. There are also significant differences in the physical behavior of materials, coating difficulty, and dispersion requirements during the molding process between ordinary tablets and microtablets. It is impossible to prepare mercaptopurine microtablets that meet quality requirements by directly referring to the formula and process of existing mercaptopurine tablets.

[0008] Therefore, it is necessary to develop a new mercaptopurine microtablet that has suitable dissolution behavior and in vivo absorption effect, and takes into account the requirements of hardness, friability and content uniformity. Summary of the Invention

[0009] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. To this end, the first aspect of the present invention provides a mercaptopurine microtablet, which has excellent in vivo pharmacokinetics, good dissolution effect, and excellent friability, hardness and content uniformity.

[0010] The second aspect of the present invention provides a method for preparing the above-mentioned mercaptopurine micro-tablets.

[0011] The third aspect of the present invention provides an application of the above-mentioned mercaptopurine micro-tablet.

[0012] According to an embodiment of the first aspect of the present invention, there is provided a mercaptopurine micro-tablet, which is circular, has a diameter of 2 mm to 3 mm, and a diameter-to-thickness ratio of 0.85 to 1.15; and comprises the following components calculated in parts by weight:

[0013] 15-30 parts of mercaptopurine API; 40-80 parts of filler; 3-8 parts of binder; 1-3 parts of lubricant; 1-3 parts of disintegrant;

[0014] Among them, the D90 particle size of the mercaptopurine raw material is 40μm~125μm; the D50 particle size of the mercaptopurine raw material is 5μm~35μm.

[0015] According to some preferred embodiments of the present invention, the mercaptopurine micro-tablet is preferably planar, planar with beveled edges, planar with arced edges or biconvex.

[0016] According to some preferred embodiments of the present invention, the diameter-to-thickness ratio of the mercaptopurine micro-sheet is preferably 0.9-1.1; for example, it can be 0.9, 0.95, 1, 1.05, 1.1 or a sub-range consisting of any two values therein.

[0017] According to some preferred embodiments of the present invention, in the mercaptopurine microtablets, the content of mercaptopurine raw material is preferably 20 to 30 parts by weight, for example, including 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, 30 parts or a sub-range consisting of any two parts thereof.

[0018] According to some preferred embodiments of the present invention, the D90 particle size of the mercaptopurine API is 40 μm to 120 μm, more preferably 40 μm to 110 μm, and most preferably 50 μm to 70 μm, for example, including 40 μm, 50 μm, 60 μm, 65 μm, 66.7 μm, 70 μm, 80 μm, 83 μm, 90 μm, 100 μm, 105 μm, 108 μm, 110 μm, 120 μm, or a subrange consisting of any two values therein.

[0019] According to some preferred embodiments of the present invention, the D50 particle size of the mercaptopurine API is 5 μm to 30 μm, more preferably 10 μm to 30 μm, and most preferably 15 μm to 30 μm, for example, including 5 μm, 10 μm, 15 μm, 15.8 μm, 16.5 μm, 17 μm, 18 μm, 19 μm, 20 μm, 30 μm, or a subrange consisting of any two values therein.

[0020] According to some preferred embodiments of the present invention, the labeled amount of the active ingredient of the mercaptopurine micro-tablet is 5 mg.

[0021] According to some preferred embodiments of the present invention, there is no particular limitation on the filler used in the mercaptopurine microtablets, preferably at least one of lactose, pregelatinized starch, mannitol, sorbitol, maltodextrin, and microcrystalline cellulose; more preferably at least one of lactose, mannitol, and microcrystalline cellulose.

[0022] According to some preferred embodiments of the present invention, for the mercaptopurine microtablets, the content of the filler used is preferably 50 to 80 parts by weight, for example, including 50 parts, 52 parts, 55 parts, 58 parts, 60 parts, 62 parts, 65 parts, 68 parts, 70 parts, 75 parts, 80 parts or a sub-range consisting of any two parts thereof.

[0023] According to some preferred embodiments of the present invention, the filler comprises lactose and microcrystalline cellulose.

[0024] According to some preferred embodiments of the present invention, the weight ratio of lactose to microcrystalline cellulose is (1~2):1, for example, including 1:1, 1.2:1, 1.4:1, 1.6:1, 1.8:1, 2:1 or a sub-range consisting of any two values therein.

[0025] According to some preferred embodiments of the present invention, there is no special limitation on the binder used in the mercaptopurine microtablets, preferably at least one of povidone, polyvinyl alcohol, hydroxypropyl methylcellulose, carbomer, hydroxypropyl cellulose, and copovidone; more preferably at least one of povidone, hydroxypropyl methylcellulose, and copovidone.

[0026] According to some preferred embodiments of the present invention, for the mercaptopurine microtablets, the content of the adhesive used is preferably 4 to 8 parts by weight, for example, including 4 parts, 5 parts, 6 parts, 7 parts, 8 parts or a sub-range consisting of any two of these values.

[0027] According to some preferred embodiments of the present invention, there is no particular limitation on the lubricant used in the mercaptopurine microtablets, and preferably at least one of magnesium stearate, calcium stearate, micropowder silica gel, and sodium stearyl fumarate.

[0028] According to some preferred embodiments of the present invention, there is no particular limitation on the disintegrant used in the mercaptopurine microtablets, preferably at least one of cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, carboxymethyl cellulose calcium, and sodium carboxymethyl starch; more preferably at least one of cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, and sodium carboxymethyl starch.

[0029] According to some preferred embodiments of the present invention, the dosage form of the mercaptopurine microtablets includes at least one of lozenges, sublingual tablets, orally disintegrating tablets, and dispersible tablets; those skilled in the art will understand that in order to prepare the mercaptopurine microtablets into the dosage form, they may also contain other necessary excipients commonly used in the art, such as flavoring agents, colorants, antioxidants, coating materials, sustained-release / controlled-release materials, etc.

[0030] According to some preferred embodiments of the present invention, the mercaptopurine micro-tablets may further contain other pharmaceutically active ingredients, which may be substances that reduce the possible side effects of mercaptopurine, such as hepatotoxicity, immunosuppression, and bone marrow suppression; or may be substances that enhance the efficacy of mercaptopurine, such as substances that have additive or even synergistic effects with it; the efficacy preferably inhibits or even treats cancer, and the other pharmaceutically active ingredients may be at least one of methotrexate, cytarabine, dihydroartemisinin, artemisinin, artesunate, artemether, and arteether.

[0031] The mercaptopurine micro-tablets according to the present invention have at least the following beneficial effects:

[0032] The thiopurine microtablets provided by the present invention have a diameter of 2 mm to 3 mm and a diameter-to-thickness ratio of 0.85 to 1.15. These microtablets solve quality problems such as fragility, cracking, and large weight variations that are easily caused by a large or small diameter-to-thickness ratio. These microtablets have excellent uniformity and good in vivo drug absorption, and are consistent in quality and efficacy with a control preparation (thiopurine tablets (50 mg) with a diameter of 9.5 mm, marketed under the trade name Purinethol).

[0033] Furthermore, for microtablets, the present invention uses a mercaptopurine raw material with a D90 particle size limited to the range of 40μm~125μm and a D50 particle size limited to the range of 5μm~35μm, and is combined with the adhesive, filler, lubricant and disintegrant of the present invention, so that the prepared mercaptopurine microtablets have the advantages of excellent dissolution behavior and in vivo absorption effect, suitable hardness, low friability and excellent content uniformity.

[0034] According to an embodiment of the second aspect of the present invention, there is provided a method for preparing the above-mentioned mercaptopurine micro-tablets, comprising the following steps:

[0035] S1. Premixing the mercaptopurine API, any other pharmaceutically active ingredients, a filler, and a binder to obtain a premix;

[0036] S2, mixing the premix and water to perform wet granulation, and then wet granulating to obtain wet granules;

[0037] S3, drying and granulating the wet granules to obtain dry granules;

[0038] S4, mixing the dry granules, lubricant and disintegrant to obtain mixed granules;

[0039] S5, compressing the total mixed granules into tablets to obtain the product.

[0040] According to some preferred embodiments of the present invention, in step S2, the parameters of the wet granulation include:

[0041] The atomizing spray gun pressure is 1 bar ~ 1.5 bar; the peristaltic pump speed is 20 rpm ~ 80 rpm; the spray speed is 0.34 kg / min ~ 0.54 kg / min.

[0042] According to some preferred embodiments of the present invention, in step S3, the moisture content of the dry particles is no more than 2%.

[0043] According to some preferred embodiments of the present invention, in step S4, the mixing time is 5 min to 8 min, for example, including 5 min, 6 min, 7 min, 8 min, or a sub-range consisting of any two values therein.

[0044] According to some preferred embodiments of the present invention, in step S4, the speed of the mixer used for mixing is 10 rpm to 50 rpm, for example, including 10 rpm, 15 rpm, 20 rpm, 25 rpm, 30 rpm, 35 rpm, 40 rpm, 45 rpm, 50 rpm, or a subrange consisting of any two values therein.

[0045] According to some preferred embodiments of the present invention, in step S4, the particle size distribution of the total mixed particles is as follows: the weight proportion of particles with a diameter less than 150 μm is 20% to 75%; the weight proportion of particles with a diameter between 150 μm and 250 μm is 15% to 35%; and the weight proportion of particles with a diameter greater than 250 μm is 5% to 55%.

[0046] According to some preferred embodiments of the present invention, in step S4, the particle size distribution of the total mixed particles is as follows: the weight proportion of particles with a diameter less than 150 μm is 55% to 75%; the weight proportion of particles with a diameter between 150 μm and 250 μm is 20% to 35%; and the weight proportion of particles with a diameter greater than 250 μm is 5% to 15%.

[0047] According to some preferred embodiments of the present invention, in step S4, the particle size distribution of the total mixed particles is as follows: the weight proportion of particles with a diameter less than 150 μm is 61.8%; the weight proportion of particles with a diameter between 150 μm and 250 μm is 27.3%; and the weight proportion of particles with a diameter greater than 250 μm is 10.9%.

[0048] By controlling the particle size distribution of the total mixed particles within the range of the present invention, the fluidity, content uniformity, etc. can be improved, and the mercaptopurine microtablets of the present invention can have suitable dissolution behavior.

[0049] According to some preferred embodiments of the present invention, in step S5, the tableting pressure is 12 kN to 15 kN. For example, it includes 12 kN, 13 kN, 14 kN, 15 kN, or a subrange consisting of any two values thereof. Thus, when the tableting pressure is controlled within the above range, the prepared mercaptopurine microtablets can have excellent hardness and friability.

[0050] The preparation method according to the embodiment of the present invention has at least the following beneficial effects:

[0051] The present invention first premixes a mercaptopurine API, a filler, and a binder. The resulting premix is then mixed with water for wet granulation. The wet granulation is then dried and granulated to obtain dry granules. The dry granules are then mixed with a lubricant and a disintegrant to obtain a total mixed granule. The total mixed granule is then compressed into tablets to obtain mercaptopurine microtablets. The wet granulation process using the specific steps of the present invention provides excellent in vivo pharmacokinetics, good dissolution, and excellent friability, hardness, and content uniformity. Compared to known methods, this preparation process is simpler, more environmentally friendly, and safer.

[0052] According to an embodiment of the third aspect of the present invention, there is provided a use of the mercaptopurine micro-tablet; according to a preferred embodiment of the present invention, the mercaptopurine micro-tablet is used for treating cancer.

[0053] According to some preferred embodiments of the present invention, the cancer includes choriocarcinoma, malignant hydatidiform mole, acute lymphoblastic leukemia (ALL), acute non-lymphocytic leukemia, blast crisis of chronic myeloid leukemia, lung cancer, colorectal cancer, liver cancer, breast cancer, and melanoma.

[0054] According to some preferred embodiments of the present invention, the cancer is most preferably childhood acute lymphoblastic leukemia.

[0055] In the description of the present invention, D50 refers to Dv50, and D90 refers to Dv90.

[0056] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or will be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] The present invention will be further described below with reference to the accompanying drawings and embodiments, in which:

[0058] Figure 1 A physical picture of the mercaptopurine micro-tablet prepared in Example 1 of the present invention;

[0059] Figure 2 Graph showing the blood drug concentrations of Example 1 of the present invention and the control preparation under fasting conditions. DETAILED DESCRIPTION

[0060] The instruments and equipment used in this invention, their models, main reagents and their sources are as follows:

[0061] Wet granulator, instrument model HLSG LAB;

[0062] Fluidized bed, instrument model is LGL010;

[0063] Lifting granulator, instrument model is ZLJ-125;

[0064] Mixer, instrument model is HSD100;

[0065] Rotary tablet press, instrument model is S250W;

[0066] Dissolution apparatus, instrument model: Hanson Elite8;

[0067] High performance liquid chromatography, instrument model: Waters Alliance 2695;

[0068] Friability tester, instrument model is FRV2000;

[0069] Laser particle size analyzer, instrument model is Mastersizer 3000;

[0070] Intelligent disintegration instrument, instrument model is ZB-IE.

[0071] Mercaptopurine API, produced by Zhejiang Chengyi Pharmaceutical Co., Ltd., with reference to the particle size and particle size distribution determination method of Part IV of the 2020 edition of the Chinese Pharmacopoeia (General Chapter 0982, Method 3, dry method determination), the particle size specifications of mercaptopurine API measured by laser particle size analyzer are: D90 is 66.7μm, D50 is 15.8μm; D90 is 83μm, D50 is 7.34μm; D90 is 108μm, D50 is 14.1μm; D90 is 15.7μm, D50 is 4.51μm; D90 is 22.1μm, D50 is 5.83μm; D90 is 139μm, D50 is 70.4μm;

[0072] Lactose Granulac 200, produced by: Megler GmbH & Co. KG, Germany;

[0073] Microcrystalline cellulose 101, produced by: Redenmeier & Sons;

[0074] Povidone K29 / 32, manufactured by ISP Technologies Inc.

[0075] Sodium carboxymethyl starch, produced by Anhui Shanhe Pharmaceutical Excipients Co., Ltd.

[0076] Magnesium stearate, manufacturer: Anhui Shanhe Pharmaceutical Excipients Co., Ltd.

[0077] Other reagents not mentioned here are conventional commercial reagents used in pharmaceutical preparation processing.

[0078] The present invention is further described below with reference to examples, but the examples are not intended to limit the present invention in any way.

[0079] Example 1

[0080] This embodiment provides a mercaptopurine micro-tablet, the mercaptopurine micro-tablet has an active ingredient labeled amount of 5 mg, the dosage of each component is shown in Table 1, and the preparation method is as follows:

[0081] S1. Premix the mercaptopurine API (D90 particle size of the mercaptopurine API is 66.7 μm and D50 particle size is 15.8 μm) with lactose Granulac 200, microcrystalline cellulose 101, and povidone K29 / 32 in a wet granulator. Set the stirring motor to 150 rpm and the chopping motor to 1800 rpm for 10 minutes to obtain a premix.

[0082] S2. Adjust the atomizing gun pressure to 1.2 bar, adjust the peristaltic pump speed to 50 rpm, and adjust the spray rate to between 0.34 kg / min and 0.54 kg / min. Mix the premix with water and perform wet granulation. Use a 4×4 mm square hole sieve to sieve the granules to obtain wet granules.

[0083] S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is no more than 2%, and the granules are passed through a 0.8 mm sieve to obtain dry granules;

[0084] S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 minutes at a mixer speed of 15 rpm to obtain mixed granules.

[0085] The particle size distribution of the total mixed particles is as follows:

[0086] The weight proportion of particles with a size of less than 150 μm is 61.8%; the weight proportion of particles with a size of 150 μm to 250 μm is 27.3%; and the weight proportion of particles with a size of more than 250 μm is 10.9%.

[0087] S5. Tablets were pressed using a 9-point punch with a diameter of 3 mm. The main pressing force of the tablet press was 15 kN and the speed of the tablet press was 30 rpm. The resulting microtablets had a diameter of 3.0 mm and a thickness of 3.0 mm.

[0088] The mercaptopurine micro-tablets prepared in this embodiment are as follows Figure 1 shown.

[0089] Example 2

[0090] This embodiment provides a mercaptopurine micro-tablet, the dosage of which is shown in Table 1, and the preparation method thereof is as follows:

[0091] S1. Premix the mercaptopurine API (D90 particle size of the mercaptopurine API is 83 μm and D50 particle size is 7.34 μm) with lactose Granulac 200, microcrystalline cellulose 101, and povidone K29 / 32 in a wet granulator. Set the stirring motor to 150 rpm and the chopping motor to 1800 rpm for 10 minutes to obtain a premix.

[0092] S2. Adjust the atomizing gun pressure to 1.2 bar, adjust the peristaltic pump speed to 50 rpm, and adjust the spray rate to between 0.34 kg / min and 0.54 kg / min. Mix the premix with water and perform wet granulation. Use a 4×4 mm square hole sieve to sieve the granules to obtain wet granules.

[0093] S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is no more than 2%, and the granules are passed through a 0.8 mm sieve to obtain dry granules;

[0094] S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 minutes at a mixer speed of 15 rpm to obtain mixed granules.

[0095] S5. Tablets were pressed using a 9-point punch with a diameter of 3 mm. The main pressing force of the tablet press was 15 kN and the speed of the tablet press was 30 rpm. The resulting microtablets had a diameter of 3.0 mm and a thickness of 3.0 mm.

[0096] Example 3

[0097] This embodiment provides a mercaptopurine micro-tablet, the dosage of which is shown in Table 1, and the preparation method thereof is as follows:

[0098] S1. Premix the mercaptopurine API (D90 particle size of the mercaptopurine API is 108 μm and D50 particle size is 14.1 μm) with lactose Granulac 200, microcrystalline cellulose 101, and povidone K29 / 32 in a wet granulator. Set the stirring motor to 150 rpm and the chopping motor to 1800 rpm for 10 minutes to obtain a premix.

[0099] S2. Adjust the atomizing gun pressure to 1.2 bar, adjust the peristaltic pump speed to 50 rpm, and adjust the spray rate to between 0.34 kg / min and 0.54 kg / min. Mix the premix with water and perform wet granulation. Use a 4×4 mm square hole sieve to sieve the granules to obtain wet granules.

[0100] S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is no more than 2%, and the granules are passed through a 0.8 mm sieve to obtain dry granules;

[0101] S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 minutes at a mixer speed of 15 rpm to obtain mixed granules.

[0102] S5. Tablets were pressed using a 9-point punch with a diameter of 3 mm. The main pressing force of the tablet press was 15 kN and the speed of the tablet press was 30 rpm. The resulting microtablets had a diameter of 3.0 mm and a thickness of 3.0 mm.

[0103] Example 4

[0104] This embodiment provides a mercaptopurine micro-tablet, the dosage of which is shown in Table 1, and the preparation method thereof is as follows:

[0105] S1. Premix the mercaptopurine API (D90 particle size of the mercaptopurine API is 66.7 μm and D50 particle size is 15.8 μm) with lactose Granulac 200, microcrystalline cellulose 101, and povidone K29 / 32 in a wet granulator. Set the stirring motor to 150 rpm and the chopping motor to 1800 rpm for 10 minutes to obtain a premix.

[0106] S2. Adjust the atomizing gun pressure to 1.2 bar, adjust the peristaltic pump speed to 50 rpm, and adjust the spray rate to between 0.34 kg / min and 0.54 kg / min. Mix the premix with water and perform wet granulation. Use a 5×5 mm square hole sieve to sieve the granules to obtain wet granules.

[0107] S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is no more than 2%, and the granules are passed through a 0.8 mm sieve to obtain dry granules;

[0108] S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 minutes at a mixer speed of 15 rpm to obtain mixed granules.

[0109] The particle size distribution of the total mixed particles is as follows:

[0110] The weight proportion of particles with a size of less than 150 μm is 21.3%; the weight proportion of particles with a size of 150 μm to 250 μm is 24.4%; and the weight proportion of particles with a size of more than 250 μm is 54.3%.

[0111] S5. Tablets were pressed using a 9-point punch with a diameter of 3 mm. The main pressing force of the tablet press was 15 kN and the speed of the tablet press was 30 rpm. The resulting microtablets had a diameter of 3.0 mm and a thickness of 3.0 mm.

[0112] Examples 5-7

[0113] Examples 5 to 7 provide a series of mercaptopurine microtablets, the component amounts of which are shown in Table 1, and the preparation method is the same as that of Example 1.

[0114] Table 1

[0115]

[0116] Comparative Example 1

[0117] This comparative example provides a mercaptopurine micro-tablet, the dosage of which is the same as that of Example 1, and the preparation method thereof is as follows:

[0118] S1. Premix the mercaptopurine API (D90 particle size of the mercaptopurine API is 15.7 μm and D50 particle size is 4.51 μm) with lactose Granulac 200, microcrystalline cellulose 101, and povidone K29 / 32 in a wet granulator. Set the stirring motor to 150 rpm and the chopping motor to 1800 rpm for 10 minutes to obtain a premix.

[0119] S2. Adjust the atomizing gun pressure to 1.2 bar, adjust the peristaltic pump speed to 50 rpm, and adjust the spray rate to between 0.34 kg / min and 0.54 kg / min. Mix the premix with water and perform wet granulation. Use a 4×4 mm square hole sieve to sieve the granules to obtain wet granules.

[0120] S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is no more than 2%, and the granules are passed through a 0.8 mm sieve to obtain dry granules;

[0121] S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 minutes at a mixer speed of 15 rpm to obtain mixed granules.

[0122] S5. Tablets were pressed using a 9-point punch with a diameter of 3 mm. The main pressing force of the tablet press was 15 kN and the speed of the tablet press was 30 rpm. The resulting microtablets had a diameter of 3.0 mm and a thickness of 3.0 mm.

[0123] Comparative Example 2

[0124] This comparative example provides a mercaptopurine micro-tablet, the components and dosages of which are the same as those in Example 1, and the preparation method thereof is as follows:

[0125] S1. Premix the mercaptopurine API (D90 particle size of the mercaptopurine API is 22.1 μm and D50 particle size is 5.83 μm) with lactose Granulac 200, microcrystalline cellulose 101, and povidone K29 / 32 in a wet granulator. Set the stirring motor to 150 rpm and the chopping motor to 1800 rpm for 10 minutes to obtain a premix.

[0126] S2. Adjust the atomizing gun pressure to 1.2 bar, adjust the peristaltic pump speed to 50 rpm, and adjust the spray rate to between 0.34 kg / min and 0.54 kg / min. Mix the premix with water and perform wet granulation. Use a 4×4 mm square hole sieve to sieve the granules to obtain wet granules.

[0127] S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is no more than 2%, and the granules are passed through a 0.8 mm sieve to obtain dry granules;

[0128] S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 minutes at a mixer speed of 15 rpm to obtain mixed granules.

[0129] S5. Tablets were pressed using a 9-point punch with a diameter of 3 mm. The main pressing force of the tablet press was 15 kN and the speed of the tablet press was 30 rpm. The resulting microtablets had a diameter of 3.0 mm and a thickness of 3.0 mm.

[0130] Comparative Example 3

[0131] This comparative example provides a mercaptopurine micro-tablet, the components and dosages of which are the same as those in Example 1, and the preparation method thereof is as follows:

[0132] S1. Premix the mercaptopurine API (D90 particle size of the mercaptopurine API is 139 μm and D50 particle size is 70.4 μm) with lactose Granulac 200, microcrystalline cellulose 101, and povidone K29 / 32 in a wet granulator. Set the stirring motor to 150 rpm and the chopping motor to 1800 rpm for 10 minutes to obtain a premix.

[0133] S2. Adjust the atomizing gun pressure to 1.2 bar, adjust the peristaltic pump speed to 50 rpm, and adjust the spray rate to between 0.34 kg / min and 0.54 kg / min. Mix the premix with water and perform wet granulation. Use a 4×4 mm square hole sieve to sieve the granules to obtain wet granules.

[0134] S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is no more than 2%, and the granules are passed through a 0.8 mm sieve to obtain dry granules;

[0135] S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 minutes at a mixer speed of 15 rpm to obtain mixed granules.

[0136] S5. Tablets were pressed using a 9-point punch with a diameter of 3 mm. The main pressing force of the tablet press was 15 kN and the speed of the tablet press was 30 rpm. The resulting microtablets had a diameter of 3.0 mm and a thickness of 3.0 mm.

[0137] Comparative Example 4

[0138] This comparative example provides a mercaptopurine micro-tablet, the dosage of which is shown in Table 2.

[0139] Table 2

[0140]

[0141] The preparation method is as follows:

[0142] 1. Place mercaptopurine, lactose, microcrystalline cellulose and sodium carboxymethyl starch in a wet granulator for premixing, with the bottom paddle at 300 rpm and the side paddle at 0 rpm, and mix for 10 minutes to obtain a premixed material;

[0143] 2. Slowly pour the ethanol solution of povidone into a wet granulator and mix with the premixed material. Granulate for 1 minute with a bottom paddle at 300 rpm and a side paddle at 1000 rpm. Then, wet granulate the mixture and pass it through a 40-mesh sieve to obtain wet granules. The concentration of povidone in the ethanol solution of povidone is 5 wt%.

[0144] 3. Dry the wet granules by fluidized bed drying method (the inlet air temperature is 50℃, and the material temperature does not exceed 50℃). After drying, pass through a 40-mesh sieve to obtain dry granules.

[0145] 4. Mix the dry granules with magnesium stearate to obtain the total mixed granules.

[0146] 5. Use a 3 mm diameter, 9-point punch to compress the tablets. The punch pressure is 6.5 kN and the tablet press speed is 14.5 rpm.

[0147] Performance Testing

[0148] 1. Dissolution test

[0149] The mercaptopurine microtablets prepared in Examples 1 to 4 and Comparative Examples 1 to 4, as well as a 9.5 mm diameter mercaptopurine tablet (50 mg) control preparation with the trade name Purinethol, were respectively taken. The dissolution test was performed using a paddle method with a medium volume of 900 mL, a rotation speed of 50 revolutions per minute, and 0.1 mol / L hydrochloric acid as the dissolution medium, in accordance with the dissolution test method of Part IV of the 2020 edition of the Chinese Pharmacopoeia (General Chapter 0931, Method 2). The results of the dissolution test are shown in Table 3.

[0150] Table 3

[0151]

[0152] Mercaptopurine is a cytotoxic drug. If the dissolution rate is too fast, the child will be exposed to high concentrations of the drug, which may cause obvious adverse reactions, increase the safety of medication, and shorten the time to maintain the efficacy. However, when the dissolution of the drug is too slow, the ideal drug absorption effect may not be achieved. Mercaptopurine is a BCS IV drug with the characteristics of low solubility and low permeability, and the release characteristics of the tablet are usually affected by the particle size distribution of the raw material. Generally speaking, reducing the particle size of the raw material will increase the dissolution rate of the drug. Unexpectedly, when the particle size of the mercaptopurine raw material is selected by the present invention to be 40μm≤D90≤125μm and 5μm≤D50≤35μm, no matter how the particle size of the mercaptopurine raw material is adjusted, the prepared mercaptopurine micro-tablets have suitable dissolution behavior, and the cumulative dissolution at different time points has good stability and minimal variability. At the same time, when preparing the mercaptopurine microtablets of Comparative Examples 1-3, drug adhesion and aggregation occurred when the particle size of the API was too small or too large, leading to problems such as particle adsorption, poor fluidity, and uneven particle distribution. However, when the API particle size range of the present invention is selected, adhesion and aggregation are avoided and content uniformity is achieved, thereby achieving both drug safety and good quality controllability.

[0153] At the same time, the particle size of the particles after granulation during the preparation process usually affects the dissolution rate of the drug. However, in the present invention, although sieves with different pore sizes were used in the preparation of wet granules in Examples 1 and 4, there was no significant difference in the dissolution effect of the mercaptopurine microtablets obtained.

[0154] It can be seen that the present invention selects a suitable particle size of the mercaptopurine raw material so that the mercaptopurine microtablets prepared by the present invention have a suitable dissolution effect and excellent processability.

[0155] The control preparation dissolves slowly in a 0.1 mol / L hydrochloric acid medium, with the dissolution rate at 15 min as low as 43%. The dissolution behavior of each example and the control preparation is significantly different. In comparison, the dissolution behavior of Comparative Examples 3 and 4 is closer to that of the control preparation and is slower.

[0156] 2. Disintegration time test

[0157] The disintegration time of the mercaptopurine microtablets prepared in Example 1 and Example 4 was investigated using an intelligent disintegration instrument, according to the disintegration time test method (General Rule 0921) in Part IV of the 2020 edition of the Chinese Pharmacopoeia. The results of the disintegration time test are shown in Table 4.

[0158] Table 4

[0159]

[0160] As can be seen from the data in Table 4, the disintegration time of Examples 1 and 4 both meet the relevant requirements of the Pharmacopoeia.

[0161] 3. Hardness and friability test

[0162] The pressure required to break a microtablet during tableting corresponds to its hardness. Mercaptopurine microtablets prepared in Example 1 and Comparative Example 4 were tested for friability using a friability tester, following the 2020 edition of the Chinese Pharmacopoeia, Part IV, Tablet Friability Test Method (General Rule 0923). The results of the hardness and friability tests are shown in Table 5.

[0163] Table 5

[0164]

[0165] As shown in Table 5, when the diameter of the mercaptopurine microtablets is controlled at 3 mm and the diameter-to-thickness ratio is approximately 1, the friability of Example 1 can be reduced to 0.03% and the hardness is approximately 30 N. Compared with Comparative Example 4, Example 1 has a higher hardness but a much lower friability. In other words, the mercaptopurine microtablets of Example 1 achieve both hardness and friability. Thus, the present invention provides mercaptopurine microtablets with superior formulation performance.

[0166] 4. Content uniformity determination

[0167] The mercaptopurine microtablets prepared in Example 1 of the present invention and Comparative Example 4 were respectively taken and the content uniformity was investigated according to the content uniformity test method (General Rule 0941) of Part IV of the 2020 edition of the Chinese Pharmacopoeia. The results of the content uniformity test are shown in Table 6.

[0168] Table 6

[0169]

[0170] As can be seen from Table 6, the content uniformity of Example 1 of the present invention is much better than that of Comparative Example 4, and the technical solution of the present invention shows more excellent formulation characteristics.

[0171] 5. In vivo pharmacokinetic experiments

[0172] In vivo pharmacokinetic parameters of Example 1 and Comparative Example 3 were compared with those of a 50mg thiopurine tablet (9.5mm diameter) control formulation, marketed as Purinethol. A single-center, randomized, open-label, single-dose, two-formulation, four-period, two-sequence, fully replicated crossover design was used for a human bioequivalence study with fasting administration. Pharmacokinetic data for Example 1, Comparative Example 3, and the control formulation are shown in Table 7 below. The plasma drug concentration curves for Example 1 and the control formulation under fasting conditions are shown in Table 7. Figure 2 In Table 7, T represents Example 1 and R represents the control preparation.

[0173] Table 7

[0174]

[0175] From the data in Table 3 above, it can be seen that the in vitro dissolution behavior of Comparative Example 3 is closest to that of the control formulation. However, from the data in Table 7, it can be seen that when the bioequivalence is evaluated by in vivo pharmacokinetic experiments, the absorption rate and absorption extent of Comparative Example 3 and the control formulation are very different, and they are not bioequivalent. 0-t and AUC 0-∞ The pharmacokinetic parameters were evaluated for bioequivalence using the average bioequivalence method (ABE), and the 90% confidence interval of the least squares geometric mean ratio fell within the judgment range of 80.00% to 125.00%; the C of Example 1 and the control preparation were max Pharmacokinetic parameters were evaluated for bioequivalence using the reference formulation-corrected average bioequivalence method (RSABE), with the least squares geometric mean ratio falling within the range of 80.00%-125.00%, and the upper limit of the one-sided 95% confidence interval was ≤ 0. These results demonstrate that, under fasting conditions, Example 1 and the reference formulation were bioequivalent in terms of both rate and extent of absorption.

[0176] It can be seen that for mercaptopurine microtablets, the conventional approach of using in vitro dissolution behavior to guide bioequivalence studies cannot achieve the expected bioequivalence. The in vitro dissolution behavior of conventional tablets cannot simply be used to guide and indirectly evaluate the in vivo release of the microtablets. In other words, mercaptopurine microtablets have an in vivo release profile that is different from that of conventional tablets. The present invention achieves bioequivalence between the technical solution of the present invention and the control formulation by controlling the particle size, formulation, and process of the API.

[0177] While the above description is in conjunction with the embodiments of the present invention, the present invention is not limited to the aforementioned embodiments. Various modifications may be made within the scope of knowledge possessed by a person skilled in the art without departing from the spirit of the present invention. Furthermore, features of different embodiments and comparative examples of the present invention may be combined with one another unless they conflict.

Claims

1. A mercaptopurine microtablet, wherein the microtablet is circular, has a diameter of 2 mm to 3 mm, and a diameter-to-thickness ratio of 0.85 to 1.15; It is characterized in that The mercaptopurine micro-tablets include the following components calculated by weight: 15-30 parts of mercaptopurine API; 40-80 parts of filler; 4-8 parts of binder; 1-3 parts of lubricant; 1-3 parts of disintegrant; Wherein, the D90 particle size of the mercaptopurine raw material is 40 μm~120 μm; the D50 particle size is 5 μm~30 μm; The filler is selected from at least one of lactose and microcrystalline cellulose; The binder is selected from povidone; The disintegrant is selected from sodium carboxymethyl starch.

2. The mercaptopurine micro-sheet according to claim 1, wherein The D90 particle size of the mercaptopurine API is 40 μm to 110 μm; And / or, the D50 particle size of the mercaptopurine bulk drug is 15 μm to 30 μm.

3. The mercaptopurine micro-sheet according to claim 1 or 2, wherein The lubricant includes at least one of magnesium stearate, calcium stearate, micropowder silica gel, and sodium stearyl fumarate.

4. The mercaptopurine micro-sheet according to claim 1 or 2, wherein The micro-sheet also includes other pharmaceutically active ingredients, and the other pharmaceutically active ingredients are selected from at least one of methotrexate, cytarabine, dihydroartemisinin, artemisinin, artesunate, artemether, and arteether.

5. A method for preparing the mercaptopurine micro-tablet according to any one of claims 1 to 4, characterized in that: The steps include: S1. Premixing the mercaptopurine API, any other pharmaceutically active ingredients, a filler, and a binder to obtain a premix; S2, mixing the premix and water to perform wet granulation, and then wet granulating to obtain wet granules; S3, drying and granulating the wet granules to obtain dry granules; S4, mixing the dry granules, lubricant and disintegrant to obtain mixed granules; S5, compressing the total mixed granules into tablets to obtain the product.

6. The method for preparing mercaptopurine micro-sheet according to claim 5, wherein In step S2, the parameters of the wet granulation include: The atomizing spray gun pressure is 1 bar ~ 1.5 bar; the peristaltic pump speed is 20 rpm ~ 80 rpm; the spray rate is 0.34 kg / min ~ 0.54 kg / min; And / or, in step S4, the particle size distribution of the total mixed particles is as follows: The weight of particles with a diameter of less than 150 μm accounts for 20% to 75%; the weight of particles with a diameter of 150 μm to 250 μm accounts for 15% to 35%; the weight of particles with a diameter of more than 250 μm accounts for 5% to 55%; And / or, in step S5, the tableting pressure is 12 kN to 15 kN.

7. Use of the mercaptopurine microtablet according to any one of claims 1 to 4 in the preparation of a drug for treating cancer.

8. The use according to claim 7, characterized in that The drug is used for choriocarcinoma, malignant hydatidiform mole, acute lymphocytic leukemia, acute non-lymphocytic leukemia, blast crisis of chronic myeloid leukemia, lung cancer, colorectal cancer, liver cancer, breast cancer, and melanoma.

9. The use according to claim 8, characterized in that The drug is used to treat childhood acute lymphoblastic leukemia.

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