Mercaptopurine micro-tablet as well as preparation method and application thereof

Abstract

The invention discloses a mercaptopurine micro-tablet as well as a preparation method and application thereof. The diameter of the mercaptopurine microchip is 2 to 3mm; the ratio of the diameter to the thickness is 0.85 to 1.15; comprising the following components in parts by weight: 15-30 parts of mercaptopurine bulk drug; 40 to 80 parts of a filling agent; 3-8 parts of an adhesive; 1-3 parts of a lubricant; 1-3 parts of a disintegrating agent; the D90 particle size of the mercaptopurine bulk drug is 40 [mu] m-125 [mu] m; the D50 particle size of the mercaptopurine bulk drug is 5-35 [mu] m. The mercaptopurine micro-tablet disclosed by the invention is excellent in in-vivo pharmacokinetic performance, good in dissolution effect and excellent in friability, hardness and content uniformity; the absorption speed and the absorption degree of the pharmaceutical composition are bioequivalent to those of a tablet Purinethol serving as a control preparation, and the pharmaceutical composition has a good application prospect in tumor treatment, especially in the aspect of acute lymphocytic leukemia of children.

Description

Technical Field

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

[0002] Mercaptopurine (English name: Mercaptopurine, chemical name: 6-purinethiol, 6-mercaptopurine, 6-Mercaptopurine, 6-MP) is a yellow crystalline powder. It is a cell cycle-specific drug that inhibits the purine synthesis pathway. Its chemical structure is similar to that of hypoxanthine, so it can competitively inhibit the transformation process of hypoxanthine. Mercaptopurine enters the body and must be converted into 6-mercaptopurine ribonucleotide by phosphoribosyltransferase in the cell before it becomes active. Mercaptopurine inhibits the synthesis and metabolism of purine nucleotides by inhibiting phosphoribosyl pyrophosphate amidotransferase (PRPP amidotransferase), which is the rate-limiting enzyme for purine synthesis. Mercaptopurine can change the synthesis and function of RNA and DNA and is a widely used anti-tumor agent.

[0003] The prior art reports that mercaptopurine has an effect 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 mercaptopurine was used to treat different malignant tumor cell lines, 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 concentration of mercaptopurine increased, the protein expression of GLT8D1 decreased, and at the same time, the protein level and protein molecular weight of PD-L1 in the cell changed significantly, the total amount of PD-L1 protein decreased significantly, the molecular weight of PD-L1 protein decreased 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-cultured with activated PBMC cells (peripheral blood mononuclear cells), the killing effect of PBMC 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, with children aged 1-4 years accounting 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 acute lymphoblastic leukemia in children, but they face many difficulties in clinical application. The only tablet size on the domestic market is 50 mg, which cannot meet the personalized dosing requirements for children. When children take the drug, the 50 mg tablet needs to be manually split, which may cause changes in the physical and chemical properties of the drug and increase the risk of contamination. For children, there is also the risk of inaccurate drug division and use, which increases the hidden dangers of safety and effectiveness. For medical operators or patients' families, since this product is a cytotoxic drug, there is also a risk of drug exposure. 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, since the specifications of the microtablets are not strictly controlled, it still cannot meet the personalized dosing requirements for children and there are problems with in vivo absorption, and the prepared microtablets also have poor dissolution effects, 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.

[0006] At the same time, microtablets cannot be simply regarded as drugs that are proportionally reduced 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 the quality requirements by directly referring to the formula and process of existing mercaptopurine tablets.

[0007] 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

[0008] 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.

[0009] The second aspect of the present invention provides a method for preparing the above-mentioned mercaptopurine micro-tablets.

[0010] The third aspect of the present invention provides an application of the above-mentioned mercaptopurine micro-tablet.

[0011] According to 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: 15 to 30 parts of mercaptopurine API; 40 to 80 parts of filler; 3 to 8 parts of binder; 1 to 3 parts of lubricant; 1 to 3 parts of disintegrant; 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.

[0012] According to some preferred embodiments of the present invention, the mercaptopurine micro-tablet is preferably planar, planar beveled, planar curved or biconvex.

[0013] According to some preferred embodiments of the present invention, the diameter-to-thickness ratio of the mercaptopurine micro-flakes is preferably 0.9-1.1; for example, it may be 0.9, 0.95, 1, 1.05, 1.1 or a sub-range consisting of any two of these values.

[0014] 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, 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 of these values.

[0015] 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, it includes 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 sub-range consisting of any two values ​​therein.

[0016] 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, it includes 5 μm, 10 μm, 15 μm, 15.8 μm, 16.5 μm, 17 μm, 18 μm, 19 μm, 20 μm, 30 μm, or a sub-range consisting of any two values ​​therein.

[0017] According to some preferred embodiments of the present invention, the labeled amount of the active ingredient in the mercaptopurine micro-tablet is 5 mg.

[0018] 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.

[0019] 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 of these values.

[0020] According to some preferred embodiments of the present invention, the filler comprises lactose and microcrystalline cellulose.

[0021] 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.

[0022] According to some preferred embodiments of the present invention, there is no particular limitation on the binder used in the mercaptopurine microtablets, preferably at least one of povidone, polyvinyl alcohol, hypromellose, carbomer, hydroxypropyl cellulose, and copovidone; more preferably at least one of povidone, hypromellose, and copovidone.

[0023] 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.

[0024] 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.

[0025] 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, calcium carboxymethyl cellulose, and sodium carboxymethyl starch; more preferably at least one of cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, and sodium carboxymethyl starch.

[0026] 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 appreciate that in order to prepare the mercaptopurine microtablets into the dosage form, other necessary excipients commonly used in the art may also be included, such as flavoring agents, colorants, antioxidants, coating materials, sustained-release / controlled-release materials, etc.

[0027] According to some preferred embodiments of the present invention, the mercaptopurine micro-tablets may further contain other pharmaceutically active ingredients, and the other pharmaceutically active ingredients may be substances that reduce 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.

[0028] The mercaptopurine micro-tablets according to the present invention have at least the following beneficial effects: The diameter of the mercaptopurine microtablet provided by the present invention is 2 mm to 3 mm, and the diameter-to-thickness ratio is 0.85 to 1.15. The quality problems of the microtablet being fragile, easy to crack, and having a large weight difference, which are easily caused when the diameter-to-thickness ratio is relatively large or small, are solved, so that the microtablet has excellent uniformity and good in vivo drug absorption, and is consistent with the quality and efficacy of a control preparation (mercaptopurine tablets (50 mg) with a diameter of 9.5 mm under the trade name of Purinethol).

[0029] Furthermore, for microtablets, the present invention uses a mercaptopurine API with a D90 particle size limited to a range of 40 μm to 125 μm and a D50 particle size limited to a range of 5 μm to 35 μm, 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, lower friability and excellent content uniformity.

[0030] According to the second aspect of the present invention, there is provided a method for preparing the above-mentioned mercaptopurine micro-tablets, comprising the following steps: 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 for 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 total mixed granules; S5, compressing the total mixed granules into tablets to obtain.

[0031] According to some preferred embodiments of the present invention, in step S2, the parameters of the wet granulation include: The pressure of the atomizing spray gun is 1bar~1.5bar; the speed of the peristaltic pump is 20rpm~80rpm; the spraying speed is 0.34kg / min~0.54kg / min.

[0032] According to some preferred embodiments of the present invention, in step S3, the moisture content of the dry particles is no higher than 2%.

[0033] 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.

[0034] 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 sub-range consisting of any two values ​​therein.

[0035] 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 size of less than 150 μm is 20% to 75%; the weight proportion of particles with a size of 150 μm to 250 μm is 15% to 35%; the weight proportion of particles with a size of more than 250 μm is 5% to 55%.

[0036] 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 size of less than 150 μm is 55% to 75%; the weight proportion of particles with a size of 150 μm to 250 μm is 20% to 35%; the weight proportion of particles with a size of more than 250 μm is 5% to 15%.

[0037] 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 size less than 150 μm is 61.8%; the weight proportion of particles with a size between 150 μm and 250 μm is 27.3%; the weight proportion of particles with a size greater than 250 μm is 10.9%.

[0038] 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.

[0039] According to some preferred embodiments of the present invention, in step S5, the tabletting pressure is 12 kN to 15 kN. For example, it includes 12 kN, 13 kN, 14 kN, 15 kN or a sub-range consisting of any two values ​​thereof. Thus, when the tabletting pressure is controlled within the above range, the prepared mercaptopurine micro-tablets can have excellent hardness and friability.

[0040] The preparation method according to the embodiment of the present invention has at least the following beneficial effects: The present invention first premixes the mercaptopurine API, filler and binder, mixes the obtained premix with water for wet granulation, granulates the wet granules, dries and granulates the wet granules to obtain dry granules, mixes the dry granules, lubricant and disintegrant to obtain total mixed granules, and then compresses the total mixed granules to obtain mercaptopurine microtablets. The process sequence of the specific steps of the present invention is used for wet granulation, and the obtained mercaptopurine microtablets have excellent in vivo pharmacokinetics, good dissolution effect, excellent friability, hardness and content uniformity. Compared with the known method, the preparation process is simpler, green and environmentally friendly, and has high safety.

[0041] According to an embodiment of the third aspect of the present invention, there is provided the application of the mercaptopurine micro-tablet; according to a preferred embodiment of the present invention, the mercaptopurine micro-tablet is used for treating cancer.

[0042] According to some preferred embodiments of the present invention, the cancer includes choriocarcinoma, malignant hydatidiform mole, acute lymphocytic leukemia (ALL), acute non-lymphocytic leukemia, blast crisis of chronic myeloid leukemia, lung cancer, colorectal cancer, liver cancer, breast cancer, and melanoma.

[0043] According to some preferred embodiments of the present invention, the cancer is most preferably childhood acute lymphoblastic leukemia.

[0044] In the description of the present invention, D50 refers to Dv50, and D90 refers to Dv90.

[0045] Additional aspects and advantages of the present invention will be given 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

[0046] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 A physical picture of the mercaptopurine micro-tablet prepared in Example 1 of the present invention; Figure 2 The graph is a blood drug concentration curve of Example 1 of the present invention and the control preparation under fasting conditions. DETAILED DESCRIPTION

[0047] The instruments and equipment used in the present invention, their models, main reagents and their sources are as follows: Wet granulator, instrument model is HLSG LAB; Fluidized bed, instrument model is LGL010; Lifting granulator, the instrument model is ZLJ-125; Mixer, instrument model is HSD100; Rotary tablet press, instrument model is S250W; Dissolution apparatus, the instrument model is Hanson Elite8; High performance liquid chromatography, instrument model: Waters Alliance 2695; Friability tester, instrument model is FRV2000; Laser particle size analyzer, instrument model is Mastersizer 3000; Intelligent disintegration instrument, instrument model is ZB-IE.

[0048] Mercaptopurine API, production unit: 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 Rule 0982 Method 3 Dry 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; Lactose Granulac 200, produced by: Megler GmbH & Co. KG; Microcrystalline cellulose 101, produced by: Redenmeier & Sons; Povidone K29 / 32, produced by ISP Technologies Inc.; Sodium carboxymethyl starch, produced by Anhui Shanhe Pharmaceutical Excipients Co., Ltd. Magnesium stearate, manufacturer: Anhui Shanhe Pharmaceutical Excipients Co., Ltd.

[0049] Other reagents not mentioned here are conventional commercial reagents used in pharmaceutical preparation processing.

[0050] The present invention is further described below by way of examples, but the examples are not intended to limit the present invention in any way.

[0051] Example 1 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: S1. Put mercaptopurine API (D90 particle size of mercaptopurine API is 66.7 μm, D50 particle size is 15.8 μm), lactose Granulac 200, microcrystalline cellulose 101 and povidone K29 / 32 into a wet granulator for premixing, set the stirring motor to 150 rpm, the chopping motor to 1800 rpm, mix for 10 min, and obtain a premix; S2, adjusting the pressure of the atomizing spray gun to 1.2 bar, adjusting the speed of the peristaltic pump to 50 rpm, adjusting the spraying speed to between 0.34 kg / min and 0.54 kg / min, mixing the premix with water, performing wet granulation, and using a 4×4 mm square hole sieve to granulate to obtain wet granules; S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is not higher than 2%, and the dry granules are obtained by a lifting granulator using a 0.8 mm round hole sieve; S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 min at a mixer speed of 15 rpm to obtain total mixed granules.

[0052] The particle size distribution of the total mixed particles is as follows: The weight proportion of particles with a size of less than 150μm is 61.8%; the weight proportion of particles with a size of 150μm to 250μm is 27.3%; and the weight proportion of particles with a size of more than 250μm is 10.9%.

[0053] S5. Tablets were pressed using a 3 mm diameter, 9-head punch. The main pressing force of the tablet press was 15 kN and the tablet press speed was 30 rpm. The diameter of the obtained microtablets was 3.0 mm and the thickness was 3.0 mm.

[0054] The actual mercaptopurine micro-tablets prepared in this example are as follows Figure 1 shown.

[0055] Example 2 This embodiment provides a mercaptopurine micro-tablet, the dosage of its components is shown in Table 1, and its preparation method is as follows: S1. Put mercaptopurine API (D90 particle size of mercaptopurine API is 83 μm, D50 particle size is 7.34 μm), lactose Granulac 200, microcrystalline cellulose 101 and povidone K29 / 32 into a wet granulator for premixing, set the stirring motor to 150 rpm, the chopping motor to 1800 rpm, mix for 10 min, and obtain a premix; S2, adjusting the pressure of the atomizing spray gun to 1.2 bar, adjusting the speed of the peristaltic pump to 50 rpm, adjusting the spraying speed to between 0.34 kg / min and 0.54 kg / min, mixing the premix with water, performing wet granulation, and using a 4×4 mm square hole sieve to granulate to obtain wet granules; S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is not higher than 2%, and the dry granules are obtained by a lifting granulator using a 0.8 mm round hole sieve; S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 min at a mixer speed of 15 rpm to obtain total mixed granules.

[0056] S5. Tablets were pressed using a 3 mm diameter, 9-head punch. The main pressing force of the tablet press was 15 kN and the tablet press speed was 30 rpm. The diameter of the obtained microtablets was 3.0 mm and the thickness was 3.0 mm.

[0057] Example 3 This embodiment provides a mercaptopurine micro-tablet, the dosage of its components is shown in Table 1, and its preparation method is as follows: S1. Put mercaptopurine API (D90 particle size of mercaptopurine API is 108 μm, D50 particle size is 14.1 μm), lactose Granulac 200, microcrystalline cellulose 101 and povidone K29 / 32 into a wet granulator for premixing, set the stirring motor to 150 rpm, the chopping motor to 1800 rpm, mix for 10 min, and obtain a premix; S2, adjusting the pressure of the atomizing spray gun to 1.2 bar, adjusting the speed of the peristaltic pump to 50 rpm, adjusting the spraying speed to between 0.34 kg / min and 0.54 kg / min, mixing the premix with water, performing wet granulation, and using a 4×4 mm square hole sieve to granulate to obtain wet granules; S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is not higher than 2%, and the dry granules are obtained by a lifting granulator using a 0.8 mm round hole sieve; S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 min at a mixer speed of 15 rpm to obtain total mixed granules.

[0058] S5. Tablets were pressed using a 3 mm diameter, 9-head punch. The main pressing force of the tablet press was 15 kN and the tablet press speed was 30 rpm. The diameter of the obtained microtablets was 3.0 mm and the thickness was 3.0 mm.

[0059] Example 4 This embodiment provides a mercaptopurine micro-tablet, the dosage of its components is shown in Table 1, and its preparation method is as follows: S1. Put mercaptopurine API (D90 particle size of mercaptopurine API is 66.7 μm, D50 particle size is 15.8 μm), lactose Granulac 200, microcrystalline cellulose 101 and povidone K29 / 32 into a wet granulator for premixing, set the stirring motor to 150 rpm, the chopping motor to 1800 rpm, mix for 10 min, and obtain a premix; S2, adjusting the pressure of the atomizing spray gun to 1.2 bar, adjusting the speed of the peristaltic pump to 50 rpm, adjusting the spraying speed to between 0.34 kg / min and 0.54 kg / min, mixing the premix with water, performing wet granulation, and using a 5×5 mm square hole sieve to granulate to obtain wet granules; S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is not higher than 2%, and the dry granules are obtained by a lifting granulator using a 0.8 mm round hole sieve; S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 min at a mixer speed of 15 rpm to obtain total mixed granules.

[0060] The particle size distribution of the total mixed particles is as follows: The weight proportion of particles with a size of less than 150μm is 21.3%; the weight proportion of particles with a size of 150μm to 250μm is 24.4%; and the weight proportion of particles with a size of more than 250μm is 54.3%.

[0061] S5. Tablets were pressed using a 3 mm diameter, 9-head punch. The main pressing force of the tablet press was 15 kN and the tablet press speed was 30 rpm. The diameter of the obtained microtablets was 3.0 mm and the thickness was 3.0 mm.

[0062] Embodiment 5~7 Examples 5 to 7 provide a series of mercaptopurine micro-tablets, the component amounts of which are shown in Table 1, and the preparation method is the same as that of Example 1.

[0063] Table 1

[0064] Comparative Example 1 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: S1. Put mercaptopurine API (D90 particle size of mercaptopurine API is 15.7 μm, D50 particle size is 4.51 μm), lactose Granulac 200, microcrystalline cellulose 101 and povidone K29 / 32 into a wet granulator for premixing, set the stirring motor to 150 rpm, the chopping motor to 1800 rpm, mix for 10 min, and obtain a premix; S2, adjusting the pressure of the atomizing spray gun to 1.2 bar, adjusting the speed of the peristaltic pump to 50 rpm, adjusting the spraying speed to between 0.34 kg / min and 0.54 kg / min, mixing the premix with water, performing wet granulation, and using a 4×4 mm square hole sieve to granulate to obtain wet granules; S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is not higher than 2%, and the dry granules are obtained by a lifting granulator using a 0.8 mm round hole sieve; S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 min at a mixer speed of 15 rpm to obtain total mixed granules.

[0065] S5. Tablets were pressed using a 3 mm diameter, 9-head punch. The main pressing force of the tablet press was 15 kN and the tablet press speed was 30 rpm. The diameter of the obtained microtablets was 3.0 mm and the thickness was 3.0 mm.

[0066] Comparative Example 2 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: S1. Put mercaptopurine API (D90 particle size of mercaptopurine API is 22.1 μm, D50 particle size is 5.83 μm), lactose Granulac 200, microcrystalline cellulose 101 and povidone K29 / 32 into a wet granulator for premixing, set the stirring motor to 150 rpm, the chopping motor to 1800 rpm, mix for 10 min, and obtain a premix; S2, adjusting the pressure of the atomizing spray gun to 1.2 bar, adjusting the speed of the peristaltic pump to 50 rpm, adjusting the spraying speed to between 0.34 kg / min and 0.54 kg / min, mixing the premix with water, performing wet granulation, and using a 4×4 mm square hole sieve to granulate to obtain wet granules; S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is not higher than 2%, and the dry granules are obtained by a lifting granulator using a 0.8 mm round hole sieve; S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 min at a mixer speed of 15 rpm to obtain total mixed granules.

[0067] S5. Tablets were pressed using a 3 mm diameter, 9-head punch. The main pressing force of the tablet press was 15 kN and the tablet press speed was 30 rpm. The diameter of the obtained microtablets was 3.0 mm and the thickness was 3.0 mm.

[0068] Comparative Example 3 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: S1. Put mercaptopurine API (D90 particle size of mercaptopurine API is 139 μm, D50 particle size is 70.4 μm), lactose Granulac 200, microcrystalline cellulose 101 and povidone K29 / 32 into a wet granulator for premixing, set the stirring motor to 150 rpm, the chopping motor to 1800 rpm, mix for 10 min, and obtain a premix; S2, adjusting the pressure of the atomizing spray gun to 1.2 bar, adjusting the speed of the peristaltic pump to 50 rpm, adjusting the spraying speed to between 0.34 kg / min and 0.54 kg / min, mixing the premix with water, performing wet granulation, and using a 4×4 mm square hole sieve to granulate to obtain wet granules; S3, after the wet granules are dried in a fluidized bed, the moisture content of the granules is not higher than 2%, and the dry granules are obtained by a lifting granulator using a 0.8 mm round hole sieve; S4. Mix the dry granules with magnesium stearate and sodium carboxymethyl starch for 7 min at a mixer speed of 15 rpm to obtain total mixed granules.

[0069] S5. Tablets were pressed using a 3 mm diameter, 9-head punch. The main pressing force of the tablet press was 15 kN and the tablet press speed was 30 rpm. The diameter of the obtained microtablets was 3.0 mm and the thickness was 3.0 mm.

[0070] Comparative Example 4 This comparative example provides a mercaptopurine micro-tablet, and the dosage of its components is shown in Table 2.

[0071] Table 2

[0072] The preparation method is as follows: 1. Put mercaptopurine, lactose, microcrystalline cellulose and sodium carboxymethyl starch into 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; 2. Slowly pour the ethanol solution of povidone into the wet granulator and mix with the premixed materials, granulate for 1 minute, bottom paddle 300rpm, side paddle 1000rpm; then perform wet granulation, pass through a 40-mesh sieve to obtain wet granules; wherein the concentration of povidone in the ethanol solution of povidone is 5wt%.

[0073] 3. Dry the wet granules by fluidized bed drying (the air inlet temperature is 50°C and the material temperature does not exceed 50°C). After drying, pass through a 40-mesh sieve to obtain dry granules. 4. Mix the dry granules with magnesium stearate to obtain the total mixed granules.

[0074] 5. Use a 3 mm diameter, 9-point punch to press the tablets. The punch pressure is 6.5 kN and the tablet press speed is 14.5 rpm.

[0075] Performance Testing 1. Dissolution test The mercaptopurine microtablets prepared in Examples 1 to 4 and Comparative Examples 1 to 4 and the control preparation of mercaptopurine tablets (50 mg) with a diameter of 9.5 mm under the trade name of Purinethol were taken respectively, and the dissolution test was carried out using a dissolution apparatus with a paddle method, 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. The results of the dissolution test are shown in Table 3.

[0076] Table 3

[0077] As a cytotoxic drug, mercaptopurine has a too fast dissolution rate, which will expose children to high concentrations of drugs, may produce obvious adverse reactions, increase drug safety issues, and shorten the time to maintain drug 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, which has the characteristics of low solubility-low permeability, and the release characteristics of tablets are usually affected by the particle size distribution of the raw material drug. Generally speaking, reducing the particle size of the raw material drug will increase the drug dissolution rate. Unexpectedly, when the particle size of the mercaptopurine raw material drug selected by the present invention is 40μm≤D90≤125μm and 5μm≤D50≤35μm, no matter how the particle size of the mercaptopurine raw material drug 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 will occur when the particle size of the raw material is too small or too large, resulting in problems such as particle adsorption, poor fluidity, and uneven particle distribution. When the above-mentioned raw material particle size range of the present invention is selected, neither adhesion nor aggregation will occur, and the content uniformity is good, which can take into account drug safety and has good quality controllability.

[0078] At the same time, the particle size after granulation in the preparation process usually also affects the dissolution rate of the drug. However, in the present invention, although screens with different aperture sizes were used in Example 1 and Example 4 when preparing wet granules, there was no obvious difference in the dissolution effect of the obtained mercaptopurine microtablets.

[0079] It can be seen that the present invention selects a suitable particle size of the mercaptopurine API so that the mercaptopurine microtablets prepared by the present invention have a suitable dissolution effect and excellent processability.

[0080] The control preparation dissolves slowly in 0.1 mol / L hydrochloric acid medium, and the dissolution at 15 min is as low as 43%. The dissolution behavior of each embodiment is significantly different from that of the control preparation. In comparison, the dissolution behavior of Comparative Examples 3 and 4 is closer to that of the control preparation and the dissolution is slower.

[0081] 2. Disintegration time test The thiopurine microtablets prepared in Example 1 and Example 4 were taken respectively, and the disintegration time limit test method (General Rule 0921) of Part IV of the 2020 edition of the Chinese Pharmacopoeia was used to investigate the disintegration time limit using an intelligent disintegration instrument. The results of the disintegration time limit test are shown in Table 4.

[0082] Table 4

[0083] From the data in Table 4, it can be seen that the disintegration time of Examples 1 and 4 both meet the relevant requirements of the Pharmacopoeia.

[0084] 3. Hardness and friability test The pressure that breaks the microtablets during tableting corresponds to the hardness of the corresponding microtablets. The mercaptopurine microtablets prepared in Example 1 of the present invention and Comparative Example 4 were taken respectively, and the friability test was performed using a friability tester in accordance with the friability test method for tablets in Part IV of the 2020 edition of the Chinese Pharmacopoeia (General Rule 0923). The results of the hardness test and the friability test are shown in Table 5.

[0085] Table 5

[0086] From the data in Table 5, it can be seen that the present invention controls the diameter of the mercaptopurine microtablet to 3 mm, and when the diameter-to-thickness ratio is about 1, the friability of Example 1 can be as low as 0.03%, and the hardness is about 30N. Compared with Comparative Example 4, Example 1 has a higher hardness, but a much lower friability, that is, the mercaptopurine microtablet of Example 1 can take into account both hardness and friability. It can be seen that the present invention obtains mercaptopurine microtablets with better formulation performance.

[0087] 4. Content uniformity determination The mercaptopurine microtablets prepared in Example 1 of the present invention and Comparative Example 4 were respectively taken, and the content uniformity was investigated with reference 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.

[0088] Table 6

[0089] It can be seen from Table 6 that 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.

[0090] 5. In vivo pharmacokinetic experiments The in vivo pharmacokinetic parameters of Example 1 and Comparative Example 3 were compared with the control preparation of mercaptopurine tablets (50 mg) with a diameter of 9.5 mm under the trade name Purinethol. A single-center, randomized, open, single-dose, two-preparation, four-cycle, two-sequence complete repeated crossover design was used to study the bioequivalence of fasting administration in humans. The pharmacokinetic data of Example 1, Comparative Example 3 and the control preparation are shown in Table 7 below. The blood drug concentration curves of Example 1 and the control preparation under fasting conditions are shown in Table 7 below. Figure 2 In Table 7, T represents Example 1, and R represents the control preparation.

[0091] Table 7

[0092] 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 preparation. 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 preparation are very different, and the bioequivalence is not good. 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 max The pharmacokinetic parameters were evaluated for bioequivalence using the reference preparation corrected average bioequivalence method (RSABE), and the least squares geometric mean ratio fell within the range of 80.00%-125.00%, and the upper limit of the one-sided 95% confidence interval was ≤ 0. The above results show that under fasting conditions, Example 1 and the reference preparation are bioequivalent in terms of absorption rate and absorption extent.

[0093] It can be seen that for mercaptopurine microtablets, the conventional thinking of using in vitro dissolution behavior to guide bioequivalence studies cannot achieve the expected bioequivalence, and the in vitro dissolution behavior of ordinary tablets cannot simply be used to guide and indirectly evaluate the in vivo release of microtablets, that is, mercaptopurine microtablets have an in vivo release law different from that of ordinary tablets. The present invention achieves the bioequivalence of the technical solution of the present invention and the control preparation by controlling the particle size, formula, and process of the raw material.

[0094] The above is a detailed description of the embodiments of the present invention, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the features of different embodiments and comparative examples of the present invention can be combined with each other without conflict.

Claims

1. A mercaptopurine micro-tablet, wherein the micro-tablet 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 to 30 parts of mercaptopurine API; 40 to 80 parts of filler; 3 to 8 parts of binder; 1 to 3 parts of lubricant; 1 to 3 parts of disintegrant; Among them, the D90 particle size of the mercaptopurine raw material is 40μm~125μm; the D50 particle size is 5μm~35μm.

2. The mercaptopurine micro-tablet according to claim 1, characterized in that: 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-tablet according to claim 1 or 2, characterized in that: The filler comprises at least one of lactose, pregelatinized starch, mannitol, sorbitol, maltodextrin, and microcrystalline cellulose; And / or, the binder comprises at least one of povidone, polyvinyl alcohol, hydroxypropyl methylcellulose, carbomer, hydroxypropyl cellulose, and copovidone; And / or, the lubricant includes at least one of magnesium stearate, calcium stearate, micro powder silica gel, and sodium stearyl fumarate; And / or, the disintegrant comprises at least one of cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, calcium carboxymethyl cellulose, and sodium carboxymethyl starch.

4. The mercaptopurine micro-tablet according to claim 1 or 2, characterized in that: The dosage form of the mercaptopurine micro-tablets includes at least one of lozenges, sublingual tablets, orally disintegrating tablets, and dispersible tablets.

5. The mercaptopurine micro-tablet according to claim 1 or 2, characterized in that: The micro-tablet 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.

6. A method for preparing the mercaptopurine micro-tablet according to any one of claims 1 to 5, 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 for 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 total mixed granules; S5, compressing the total mixed granules into tablets to obtain.

7. The method for preparing the mercaptopurine micro-tablet according to claim 6, characterized in that: In step S2, the parameters of the wet granulation include: The pressure of the atomizing spray gun is 1 bar ~ 1.5 bar; the speed of the peristaltic pump is 20 rpm ~ 80 rpm; the spray speed 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 is 20% to 75%; the weight of particles with a diameter of 150 μm to 250 μm is 15% to 35%; the weight of particles with a diameter of more than 250 μm is 5% to 55%; And / or, in step S5, the tableting pressure is 12 kN~15 kN.

8. Use of the mercaptopurine microtablets according to any one of claims 1 to 5 in the preparation of drugs for treating cancer.

9. The use according to claim 8, 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.

10. The use according to claim 9, characterized in that: The drug is used for childhood acute lymphoblastic leukemia.

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