A preparation method of maraviroc tablets and application thereof

Abstract

This invention belongs to the field of pharmaceutical technology, specifically relating to a method for preparing mabaloxavir tablets and its application. This invention uses lactose dodecyl ester and microcrystalline cellulose to microcrystallize mabaloxavir, reducing its particle size and increasing its specific surface area, thereby accelerating the drug's dissolution rate in vivo, improving its bioavailability, allowing the drug to exert its effects more quickly, and shortening the patient's treatment cycle. The use of lactose dodecyl ester, microcrystalline cellulose, croscarmellose sodium, and povidone K25 as excipients helps form a uniform and stable carrier solution and mixture, and also improves the tablet's formability, disintegration, and dissolution properties during tableting, improving tablet quality and stability, reducing gastrointestinal irritation, and enhancing patient medication adherence.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and particularly relates to a preparation method of marbofloxacin tablets and application thereof. BACKGROUND

[0002] Marbofloxacin is a new type of anti-influenza drug, which belongs to RNA polymerase inhibitors and can directly inhibit the synthesis of influenza virus RNA. Marbofloxacin is a prodrug which is converted into active metabolite baloxavir through hydrolysis, and marbofloxacin inhibits the endonuclease activity of polymerase acid (PA) protein (an influenza virus specific enzyme in the RNA polymerase complex required for viral gene transcription), thereby inhibiting influenza virus replication.

[0003] Marbofloxacin tablets were approved for marketing in Japan and the United States in 2018, and in China in 2021. In the same year, it was included in the national medical insurance list and belongs to medical insurance class B drugs. In 2022, it was included in the Expert Consensus on Antiviral Therapy for Adult Influenza. Marbofloxacin dry suspension was approved for marketing by FDA in November 2020, and was approved for marketing in China on December 29, 2023. However, the existing marbofloxacin tablets are expensive, and many patients have difficulty in long-term burden. Due to technical monopoly, there is no record of marbofloxacin tablet preparation in the existing technology, which limits the large-scale industrial production of marbofloxacin tablets. Therefore, it is of great practical significance to develop a more efficient, more environmentally friendly and more economical preparation process. SUMMARY

[0004] The purpose of the present application is to provide a preparation method of marbofloxacin tablets and application thereof.

[0005] The present application provides a preparation method of marbofloxacin tablets, which comprises the following steps:

[0006] Marbofloxacin raw materials are completely dissolved using ethanol to obtain a marbofloxacin solution; lactose dodecyl ester and microcrystalline cellulose are mixed to form a carrier solution by adding water; the marbofloxacin solution is slowly added to the carrier solution, stirred uniformly, and then ultrasonically treated for 10-30 min. After removing the solvent by rotary evaporation, microcrystalline marbofloxacin is obtained; the microcrystalline marbofloxacin is uniformly mixed with croscarmellose sodium, povidone K25 and sodium stearyl fumarate to obtain a mixture, and the mixture is compressed into tablets to obtain marbofloxacin coated tablets.

[0007] Preferably, the amount of each substance is 15-25 parts by mass of baloxavir marboxil raw material, 5-15 parts by mass of lactosyl dodecyl ester, 5-15 parts by mass of cross-linked sodium carboxymethyl cellulose, 5-15 parts by mass of povidone K25, 20-40 parts by mass of microcrystalline cellulose, 0.5-2 parts by mass of sodium stearyl fumarate, 5-15 parts by mass of hydroxypropyl methyl cellulose, 0.5-2 parts by mass of talc, and 0.5-2 parts by mass of titanium dioxide.

[0008] Preferably, the purity of baloxavir in the baloxavir marboxil raw material is greater than or equal to 99.0%.

[0009] Preferably, the preparation method of the lactosyl dodecyl ester comprises the following steps: mixing lactose and dodecyl methyl ester according to a molar ratio of 1:1-1.5, adding 1%-5% of the total moles of the reactants of sulfuric acid, and reacting at 60-90°C for 2-4h to obtain lactosyl dodecyl ester.

[0010] Preferably, the concentration of lactosyl dodecyl ester in the carrier solution is 5%wt-15%wt.

[0011] Preferably, the ultrasonic conditions are 50-100W, 20-40kHz ultrasonic for 15-20min, and each ultrasonic is 1-2min, and the interval is 30-60s.

[0012] Preferably, the rotary evaporation conditions are 40-50°C, 50-150rpm evaporation for 1-1.5h.

[0013] Preferably, the specification of the baloxavir marboxil tablet is 20mg / tablet or 40mg / tablet.

[0014] The application also provides the use of the above-mentioned technical solution in the preparation of anti-influenza drugs.

[0015] The application has the following beneficial effects:

[0016] Baloxavir marboxil, chemical formula C 27 H 23 F2N3O7S, structural formula as follows, white to light yellowish white powder, easily soluble in dimethyl sulfoxide, slightly soluble in acetonitrile, not easily soluble in methanol or ethanol (99.5), and almost insoluble in water. The application adopts lactosyl dodecyl ester and microcrystalline cellulose to microcrystallize baloxavir marboxil, which can reduce the particle size and increase the specific surface area of baloxavir marboxil, thereby accelerating the dissolution rate of the drug in the body, improving the bioavailability of the drug, making the drug act more quickly, and shortening the treatment cycle of the patient.

[0017]

[0018] The application selects lactose dodecyl ester, microcrystalline cellulose, crosslinked sodium carboxymethyl cellulose, povidone K25 and other excipients, and limits the dosage range thereof, which not only helps to form a uniform and stable carrier solution and mixture, but also improves the tablet forming property, disintegrating property and dissolution property of the tablet during the tablet pressing process, improves the quality and stability of the tablet, reduces the irritation of the drug in the gastrointestinal tract, and enhances the medication compliance of patients. BRIEF DESCRIPTION OF DRAWINGS

[0019] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the drawings needed to be used in the embodiments will be briefly introduced below.

[0020] Figure 1 The dissolution curve of the marboxalvir tablet with a specification of 20mg;

[0021] Figure 2 The dissolution curve of the marboxalvir tablet with a specification of 40mg. DETAILED DESCRIPTION

[0022] In order to further illustrate the application, a coffee acid taste control method provided by the application is described in detail below in combination with the drawings and embodiments, but they should not be understood as limiting the protection scope of the application.

[0023] In the embodiments of the application, the marboxalvir tablet imported by Roche Pharma (Schweiz) AG in China is used as a reference preparation, with a trade name of Xofluza (Xofluza) and a specification of 40mg.

[0024] Example 1

[0025] Lactose and dodecyl methyl ester are mixed according to a molar ratio of 1:1.2, 2% of sulfuric acid of the total moles of the reactants is added, and the reaction is carried out at 80℃ for 2h. After the reaction is completed, the generated methanol and unreacted lactose are removed by water washing, and then lactose dodecyl ester is further purified by vacuum evaporation.

[0026] Take 20 parts of marboxalvir raw material, 10 parts of lactose dodecyl ester, 10 parts of crosslinked sodium carboxymethyl cellulose, 5 parts of povidone K25, 20 parts of microcrystalline cellulose, 1 part of sodium stearyl fumarate, 8 parts of hydroxypropyl methyl cellulose, 1 part of talc and 1 part of titanium dioxide by mass fraction for standby.

[0027] The maraviroc raw material is completely dissolved using ethanol to obtain a maraviroc solution; the lactose dodecyl ester and microcrystalline cellulose are mixed to form a carrier solution; the maraviroc solution is slowly added to the carrier solution, stirred uniformly, then ultrasonically treated for 15 min, and after the solvent is removed by rotary evaporation, microcrystallized maraviroc is obtained; the microcrystallized maraviroc is uniformly mixed with croscarmellose sodium, povidone K25, and sodium stearyl fumarate to obtain a mixture, and the mixture is tableted to obtain maraviroc coated tablets.

[0028] Example 2

[0029] The lactose and dodecyl methyl ester are mixed at a molar ratio of 1:1.5, 2% of sulfuric acid based on the total moles of reactants is added, and the reaction is carried out at 70°C for 3 h. After the reaction is completed, the generated methanol and unreacted lactose are removed by water washing, and then further purified by vacuum evaporation to obtain lactose dodecyl ester.

[0030] The maraviroc raw material 25 parts, lactose dodecyl ester 15 parts, croscarmellose sodium 10 parts, povidone K25 5 parts, microcrystalline cellulose 20 parts, sodium stearyl fumarate 1 part, hydroxypropyl methyl cellulose 8 parts, talc 2 parts, and titanium dioxide 2 parts are weighed in parts by mass for standby use.

[0031] The maraviroc raw material is completely dissolved using ethanol to obtain a maraviroc solution; the lactose dodecyl ester and microcrystalline cellulose are mixed to form a carrier solution; the maraviroc solution is slowly added to the carrier solution, stirred uniformly, then ultrasonically treated for 20 min, and after the solvent is removed by rotary evaporation, microcrystallized maraviroc is obtained; the microcrystallized maraviroc is uniformly mixed with croscarmellose sodium, povidone K25, and sodium stearyl fumarate to obtain a mixture, and the mixture is tableted to obtain maraviroc coated tablets.

[0032] Example 3

[0033] The lactose and dodecyl methyl ester are mixed at a molar ratio of 1:1, 2% of sulfuric acid based on the total moles of reactants is added, and the reaction is carried out at 80°C for 2 h. After the reaction is completed, the generated methanol and unreacted lactose are removed by water washing, and then further purified by vacuum evaporation to obtain lactose dodecyl ester.

[0034] The maraviroc raw material 15 parts, lactose dodecyl ester 5 parts, croscarmellose sodium 10 parts, povidone K25 8 parts, microcrystalline cellulose 20 parts, sodium stearyl fumarate 1 part, hydroxypropyl methyl cellulose 5 parts, talc 0.5 parts, and titanium dioxide 0.5 parts are weighed in parts by mass for standby use.

[0035] The maraviroc raw material is completely dissolved using ethanol to obtain a maraviroc solution; the lactose dodecyl ester and microcrystalline cellulose are mixed to form a carrier solution by adding water; the maraviroc solution is slowly added to the carrier solution, stirred uniformly, then ultrasonically treated for 10 min, and after the solvent is removed by rotary evaporation, the microcrystallized maraviroc is obtained; the microcrystallized maraviroc, croscarmellose sodium, povidone K25 and sodium stearyl fumarate are uniformly mixed to obtain a mixture, and the mixture is tabletted to obtain the maraviroc coated tablet.

[0036] Comparative Example 1

[0037] The difference from Example 1 is that the maraviroc is not microcrystallized, and the specific process is as follows:

[0038] The lactose and dodecyl methyl ester are mixed according to a molar ratio of 1:1.2, 2% of sulfuric acid of the total moles of the reactants is added, and the reaction is carried out at 80°C for 2h. After the reaction is completed, the generated methanol and unreacted lactose are removed by water washing, and then further purified by vacuum evaporation to obtain lactose dodecyl ester.

[0039] The maraviroc raw material 20 parts, lactose dodecyl ester 10 parts, croscarmellose sodium 10 parts, povidone K25 5 parts, microcrystalline cellulose 20 parts, sodium stearyl fumarate 1 part, hydroxypropyl methyl cellulose 8 parts, talc 1 part and titanium dioxide 1 part are weighed by mass parts for standby.

[0040] The maraviroc, lactose dodecyl ester, croscarmellose sodium, povidone K25, microcrystalline cellulose and sodium stearyl fumarate are uniformly mixed to obtain a mixture, and the mixture is tabletted to obtain the maraviroc coated tablet.

[0041] Comparative Example 2

[0042] The difference from Example 1 is that the lactose is not modified, and lactose monohydrate is used instead of lactose dodecyl ester, and the specific process is as follows:

[0043] The maraviroc raw material 20 parts, lactose monohydrate 10 parts, croscarmellose sodium 10 parts, povidone K25 5 parts, microcrystalline cellulose 20 parts, sodium stearyl fumarate 1 part, hydroxypropyl methyl cellulose 8 parts, talc 1 part and titanium dioxide 1 part are weighed by mass parts for standby.

[0044] The maraviroc raw material is completely dissolved using ethanol to obtain a maraviroc solution; lactose monohydrate and microcrystalline cellulose are mixed to form a carrier solution; the maraviroc solution is slowly added to the carrier solution, stirred uniformly, then ultrasonically treated for 15 min, and after the solvent is removed by rotary evaporation, microcrystallized maraviroc is obtained; the microcrystallized maraviroc is uniformly mixed with croscarmellose sodium, povidone K25 and sodium stearyl fumarate to obtain a mixture, and the mixture is tableted to obtain maraviroc coated tablets.

[0045] Test Example 1

[0046] The bioavailability of maraviroc tablets prepared in Comparative Examples 1-3 and Comparative Examples 1-2 is tested.

[0047] Healthy adult C57BL / 6 mice are selected and then divided into five test groups, i.e., Groups 1-5, and a control group, with 6 mice in each group.

[0048] Group 1: maraviroc tablets prepared in Example 1 are used to gavage the mice, with a dosage of 0.5 mg / kg of body weight;

[0049] Group 2: maraviroc tablets prepared in Example 2 are used to gavage the mice, with a dosage of 0.5 mg / kg of body weight;

[0050] Group 3: maraviroc tablets prepared in Example 3 are used to gavage the mice, with a dosage of 0.5 mg / kg of body weight;

[0051] Group 4: maraviroc tablets prepared in Comparative Example 1 are used to gavage the mice, with a dosage of 0.5 mg / kg of body weight;

[0052] Group 5: maraviroc tablets prepared in Comparative Example 2 are used to gavage the mice, with a dosage of 0.5 mg / kg of body weight;

[0053] Control group: the mice are gavaged with a reference preparation, with a dosage of 0.5 mg / kg of body weight.

[0054] The blood of the mice is collected 6 hours after administration, and the concentration of the drug in the plasma is determined by HPLC method, and the relative bioavailability of the maraviroc tablets prepared in Groups 1-5 in the mice is calculated, with the results shown in Table 1.

[0055] Relative bioavailability F = test group C max / control group C max x 100%

[0056] Table 1: Relative bioavailability results

[0057]

[0058] From Table 1, the relative bioavailability of the tablets prepared in Example, Comparative Example 1 and Comparative Example 2 is significantly reduced, indicating that the microcrystallization of maraviroc with lactose dodecyl ester as an excipient in the present application can effectively improve the solubility and bioavailability of maraviroc.

[0059] Test Example 2

[0060] The dissolution of maraviroc tablets prepared in the present application was determined by the paddle method, and the specific steps were as follows:

[0061] For tablets with a specification of 20 mg, 900 mL of degassed pH 6.8 phosphate buffer was measured and injected into the operating container of the dissolution tester, and the dissolution medium was heated to 37°C; 0.07% cetyltrimethylammonium bromide (CTAB) was added to 900 mL of pH 6.8 phosphate buffer, and the rotation speed was set to 50 rpm. The maraviroc tablets were placed in the operating container of the dissolution tester, ensuring that the sample was completely immersed in the dissolution medium, and the dissolution tester was started, and the timing was started. The experimental time was 30 minutes. At 30 minutes, an appropriate amount of solution was drawn from the specified sampling point, and the dissolution amount of maraviroc was determined, and the results are shown in Table 2, wherein Batch number S16009-HAL, Batch number S16010-HAL, and Batch number S16016-HAL represent maraviroc coated tablets prepared in the present application, and Batch number 0001, Batch number 0002, and Batch number 0003 represent reference preparations. Figure 1

[0062] For tablets with a specification of 40 mg, 900 mL of degassed pH 6.8 phosphate buffer was measured and injected into the operating container of the dissolution tester, and the dissolution medium was heated to 37°C; 0.16% cetyltrimethylammonium bromide (CTAB) was added to 900 mL of pH 6.8 phosphate buffer, and the rotation speed was set to 50 rpm. The maraviroc tablets were placed in the operating container of the dissolution tester, ensuring that the sample was completely immersed in the dissolution medium, and the dissolution tester was started, and the timing was started. The experimental time was 30 minutes. At 30 minutes, an appropriate amount of solution was drawn from the specified sampling point, and the dissolution amount of maraviroc was determined, and the results are shown in Table 3, wherein Batch number S17002, Batch number S17003, and Batch number S17004 represent three different batches of maraviroc coated tablets prepared in the present application, respectively. Figure 2

[0063] From Table 1, the relative bioavailability of the tablets prepared in Example, Comparative Example 1 and Comparative Example 2 is significantly reduced, indicating that the microcrystallization of maraviroc with lactose dodecyl ester as an excipient in the present application can effectively improve the solubility and bioavailability of maraviroc. Figure 1 and Figure 2 ​​It can be known that the dissolution curve of the maraviroc coated tablets prepared in the embodiment of the present application is consistent with the reference preparation, which indicates that the compatibility of the excipients and the drug selected in the present application is good, and the excipients in the tablets can play a similar role to the excipients in the reference preparation, such as promoting the dispersion of the drug, increasing the dissolution rate of the drug, etc., and there is no adverse interaction with the drug to affect the dissolution of the drug; the dissolution curves of tablets of different batches also show consistency, which indicates that the preparation method provided in the present application is strictly controlled and optimized in each link of production, so that each batch of drug tablets produced can maintain a similar dissolution curve, thereby ensuring the stability and consistency of product quality, and precise process parameter control can ensure that drug tablets meeting the quality requirements can be produced in different production batches and production environments.

[0064] Test Example 2

[0065] Clinical Trial

[0066] The maraviroc tablets provided by the present application are suitable for 12-year-old and above simple type A and B influenza patients, including previously healthy patients and patients with high risk of influenza complications. A single dose of the product is taken within 48 hours after the onset of symptoms, which can be taken with or without food. The product should be avoided to be taken at the same time with dairy products, calcium fortified beverages, laxatives containing high valence cations, antacids or oral supplements (such as calcium, iron, magnesium, selenium or zinc). The product is suitable for adults and adolescents (≥12 years old), and the body weight-based dosing regimen is shown in Table 2.

[0067] Table 2 Maraviroc tablet body weight-based dosing regimen

[0068]

[0069] The test drug is the test preparation (T) and the reference preparation (R), and the bioequivalence test in the Chinese healthy subjects is carried out in a single center in an open, randomized, single dose, double cycle, double cross-over fasting and postprandial state. After signing the informed consent, the subjects will receive a series of inquiries and examinations, including: demographic data, body weight and height measurement, medical history, medication history, history of allergies, history of blood donation and blood loss, smoking and drinking, history of participating in clinical trials, history of drug abuse, physical examination, vital sign examination, 12-lead electrocardiogram examination, laboratory examination, etc. Each screening examination is carried out within 14 days before the administration in the first cycle. The researcher verifies the inclusion / exclusion criteria according to the screening results.

[0070] Inclusion criteria: (1) Subjects fully understand the purpose, nature, procedure and possible adverse reactions of the trial, voluntarily participate as subjects, and sign informed consent forms before the start of all research procedures; (2) Healthy male or female subjects aged ≥18 years; (3) Male weight ≥50.0 kg, female weight ≥45.0 kg; Body mass index (BMI) between 19.0 and 26.0 kg / m² 2 (4) Subjects voluntarily take effective contraceptive measures during the trial and within 3 months after the last administration of the study drug, and have no plans to have children, donate sperm or eggs; (5) Subjects are able to communicate well with the researchers and understand and comply with all the requirements of this study.

[0071] Exclusion criteria: (1) Those with known allergies to baloxavir and its excipients, or those with allergic constitutions (e.g., known history of allergies to two or more substances) (medical history); (2) Those with a history of chronic or severe diseases of the respiratory, circulatory, digestive, urinary, hematologic, lymphatic, endocrine, immune, mental, or nervous systems within one year prior to screening and before randomization, or those with a history of gastrointestinal diseases (including current ones) within one year prior to screening and before randomization, including chronic or active upper gastrointestinal diseases such as esophageal diseases, gastritis, duodenitis, peptic ulcers, or active gastrointestinal bleeding or gastrointestinal surgery (medical history); (3) Those who developed an acute illness within two weeks prior to screening and before randomization (medical history); (4) Those who used P-glycoprotein blockers (itraconazole), probenecid, warfarin, antacids, or oral supplements (e.g., calcium, iron, magnesium, selenium, or zinc) within the previous 28 days (medical history). (5) Those who have undergone surgery within 6 months prior to screening that, according to the investigator, would affect drug absorption, distribution, metabolism, or excretion, or those who plan to undergo surgery during the study period (inquiry); (6) Those whose vital signs, physical examination, clinical laboratory tests (complete blood count, complete urine count, blood biochemistry, coagulation tests, virology), and 12-lead electrocardiogram at the time of screening show abnormalities and whose abnormalities are clinically significant according to the investigator (examination); (7) Creatinine clearance rate <90 mL / min (creatinine clearance rate is calculated using the Cockcroft-Gault formula: CrCl=[(140-age)×weight(kg)] / [0.814×Scr(umol / L)], or CrCl=(140-age)×weight(kg) / 72×Scr(mg / dL).(8) Those who received live attenuated vaccine 2 weeks before screening to randomization, or those who need to receive live attenuated vaccine during the trial (medical history); (9) Those who used any drugs (including prescription drugs, over-the-counter drugs, Chinese herbal preparations and prescriptions, etc.) and health products 2 weeks before screening to randomization (medical history); (10) Those who were exposed to baloxavir within 3 months before administration of the study drug; (11) Those who participated in other clinical trials 3 months before screening to randomization (medical history, plagiarism check); (12) Those who donated blood 3 months before screening to randomization, or those whose total blood loss from blood donation or other reasons reached or exceeded 400 mL 6 months before screening to randomization (excluding blood loss during menstruation in women) (medical history); (13) Those who received live attenuated vaccine 48 weeks before administration each cycle. Those who cannot stop consuming caffeinated or alcoholic beverages and foods (including chocolate, tea, coffee, cola, etc.) or foods that affect drug metabolism, such as grapefruit, grapefruit products, dragon fruit, mango, pomelo, orange, star fruit, guava, etc., from 48 hours before administration to discharge from the hospital in each cycle, or those who cannot stop smoking from 48 hours before administration to discharge from the hospital in each cycle (patient visit); (14) Those with a history of alcohol abuse in the year prior to screening to randomization, i.e., an average weekly alcohol consumption of more than 2 units of alcohol (1 unit = 360 mL beer or 45 mL of 40% alcohol liquor or 150 mL of wine) (patient visit), or an alcohol breath test result greater than 0.0 mg / 100 (15) Individuals who smoke an average of more than 5 cigarettes per day from 3 months prior to screening to randomization (inquiry); (16) Individuals with a history of drug abuse (including the use of various narcotic drugs and psychotropic substances for non-medical purposes) from one year prior to screening to randomization (inquiry), or individuals who have tested positive for drug abuse screening (including: morphine, methamphetamine, ketamine, ecstasy, marijuana, etc.) (inquiry); (17) Individuals with poor vascular puncture conditions, who cannot tolerate venous puncture, or who have a history of fainting or dizziness, or who have difficulty swallowing (inquiry); (18) Individuals with hereditary fructose intolerance, glucose / galactose malabsorption syndrome, or sucrose-isomaltase deficiency (inquiry); (19) Individuals with special dietary requirements who cannot accept a uniform diet (inquiry); (20) Individuals who, according to the researcher's judgment, have other circumstances that make them unsuitable for enrollment.

[0072] In addition to the above requirements, female subjects should also be excluded if they meet the following conditions: (1) those who used oral contraceptives 30 days before screening to before randomization (medical history); (2) those who used long-acting estrogen or progestin injections (including progestin-type intrauterine devices) or implants 6 months before screening to before randomization (medical history); (3) those who had unprotected sexual intercourse with their partners 14 days before screening to before randomization (medical history); (4) those who tested positive for pregnancy or whose test results were outside the normal range or outside the range of non-pregnancy (test); (5) pregnant or lactating women (medical history).

[0073] Qualified subjects were admitted to the Phase I clinical research laboratory of the hospital on day -1 and underwent the following assessments: (1) Health status survey: latest medical history, medication history, smoking and drinking status, etc.; (2) Vital signs measurement; (3) Alcohol breath test; (4) Drug abuse screening; (5) Blood pregnancy test for women; (6) Re-verification of inclusion / exclusion criteria and final determination.

[0074] A total of 24 healthy subjects (including male and female subjects) were selected for the preliminary trial. Ten subjects (including male and female subjects) were enrolled in the fasting administration trial and ten subjects (including male and female subjects) were enrolled in the postprandial administration trial.

[0075] The formal trial enrolled 68 healthy subjects (including male and female subjects), with 34 subjects (including male and female subjects) enrolled in the fasting administration trial and 34 subjects (including male and female subjects) enrolled in the postprandial administration trial.

[0076] The test formulation (T) was mabaloxavir tablets of 40 mg prepared according to this invention, and the reference formulation (R) was commercially available mabaloxavir tablets of 40 mg (licensed by Roche Registration Ltd.). Each cycle of the study involved one dose, and healthy subjects were randomly assigned to two groups (one for fasting and one for postprandial administration), with equal numbers in each group. The washout period was 35 days. The dosing order is shown in Table 3. Fasting for at least 10 hours was required before administration or before consuming a high-fat meal. Strenuous exercise and prolonged bed rest were prohibited throughout the study. Blood samples were collected under clinical monitoring by experienced physicians and nurses. On the day of each administration cycle, subjects were required to remain upright for 2 hours after administration and avoid going to the toilet within 2 hours. If necessary, the researcher would accompany the subject. If any discomfort occurred, the research physician would promptly assess its clinical significance and provide appropriate treatment.

[0077] Table 3 Cross-dosing regimen for mabaloxavir tablets

[0078]

[0079] Fasting administration experiment

[0080] Subjects were randomly assigned to two groups, TR and RT, with equal numbers in each group. Enrolled subjects entered the Phase I clinical trial laboratory the day before each trial cycle. They ate a standard dinner and fasted for at least 10 hours before administration. On the morning of the trial, they took one tablet of the investigational drug, mabaloxavir (T, 40 mg), or one tablet of the reference drug, mabaloxavir (trade name: Sufuda®) (R, 40 mg), on an empty stomach, with approximately 240 mL of warm water. Subjects were prohibited from drinking water for 1 hour before and 1 hour after administration (except for drinking water during medication and management of adverse events). After 35 days, subjects were crossovered and began the second cycle of the study. The Phase I clinical trial laboratory drug administrator allocated the investigational drug for each phase of the study according to the randomization list.

[0081] Upper limb venous blood samples were collected at 21 time points: within 1.50 h before administration (0.00 h) and at 1 h, 2 h, 2.5 h, 3 h, 3.33 h, 3.67 h, 4 h, 4.33 h, 4.67 h, 5 h, 5.5 h, 6 h, 7 h, 8 h, 10 h, 12 h, 24 h, 36 h, 48 h, and 72 h after administration. Approximately 4 mL of blood was collected and placed into vacuum blood collection tubes containing an anticoagulant confirmed by the testing unit. After all subjects' blood samples were collected, they were transported to the analytical testing center for blood drug concentration determination according to the testing unit's transportation requirements. Specific sample pretreatment was based on the SOP issued by the final testing unit. The blood collection points for the formal trial were adjusted appropriately based on the results of the preliminary trial.

[0082] Subjects' sitting vital signs (including body temperature, pulse, and blood pressure) were measured within 1.0 h before each administration cycle and at 2.0±0.5 h, 4.0±0.5 h, 12.0±1.0 h, 24.0±1.0 h, 48.0±1.0 h after administration, and at discharge from the hospital or the group.

[0083] After all pharmacokinetic blood samples were collected in the first cycle, the subjects were allowed to leave the Phase I clinical trial center. After all pharmacokinetic blood samples were collected in the second cycle, the subjects underwent a safety check on the same day. After the check was completed (laboratory check after sample collection), they could leave the Phase I clinical trial center. Other procedures in the second cycle were the same as in the first cycle.

[0084] Postprandial Dosing Trial

[0085] A suitable number of qualified healthy subjects (including both male and female subjects) were randomly assigned to two groups: TR and RT groups, with equal numbers in each group. Enrolled subjects entered the Phase I clinical trial laboratory the day before each trial cycle. They ate a standard dinner, fasting for at least 10 hours before consuming the high-fat meal. On the morning of the trial, following the protocol, they took one tablet of the test drug, mabaloxavir (T, 40 mg), or one tablet of the reference drug, mabaloxavir (trade name: Sufuda®) (R, 40 mg), orally after the high-fat meal, with approximately 240 mL of warm water. The high-fat meal was to be consumed within 30 minutes of the start of the meal, and the test drug was to be taken 30 minutes after the start of the meal. Subjects were prohibited from drinking water for 1 hour before meals and 1 hour after drug administration (except for water consumed during drug administration, beverage formulation during the high-fat meal, and adverse event management). After 35 days, subjects were crossover-administered for the second cycle of the study. Phase I clinical trial administrators assign investigational drugs to each phase of the study based on the randomization list.

[0086] Upper limb venous blood samples were collected at 21 time points: within 1.50 h before administration (0.00 h) and at 1 h, 2 h, 2.5 h, 3 h, 3.33 h, 3.67 h, 4 h, 4.33 h, 4.67 h, 5 h, 5.5 h, 6 h, 7 h, 8 h, 10 h, 12 h, 24 h, 36 h, 48 h, and 72 h after administration. Approximately 4 mL of blood was collected and placed in vacuum blood collection tubes containing an anticoagulant confirmed by the testing unit. Sample pretreatment was the same as for the fasting test. After all subjects' blood samples were collected, they were transported to the analytical testing center for blood drug concentration determination according to the testing unit's transportation requirements. The blood collection points for the formal test were adjusted appropriately based on the results of the preliminary test.

[0087] Subjects' sitting vital signs (including body temperature, pulse, and blood pressure) were measured within 1.0 h before each administration cycle and at 2.0±0.5 h, 4.0±0.5 h, 12.0±1.0 h, 24.0±1.0 h, 48.0±1.0 h after administration, and at discharge from the hospital or the group.

[0088] After all pharmacokinetic blood samples were collected in the first cycle, the subjects were allowed to leave the Phase I clinical trial center. After all pharmacokinetic blood samples were collected in the second cycle, the subjects underwent a safety check on the same day. After the check was completed (laboratory check after sample collection), they could leave the Phase I clinical trial center. Other procedures in the second cycle were the same as in the first cycle.

[0089] Discharge: On the day the sampling ended in the second cycle, the subjects underwent physical examination, vital sign examination, electrocardiogram and relevant laboratory tests. Adverse events, concomitant medications and non-drug treatments were recorded during the trial.

[0090] Follow-up: When an adverse medical event occurs after randomization during the trial, the investigator must follow up the subject until the subject recovers, the condition improves, stabilizes, is lost to follow-up, or the event can be explained in another way.

[0091] Before statistical analysis, if any of the following situations occur, the principal investigator, sponsor, and biostatistician will make a comprehensive judgment based on factors such as the participant's completion of the trial and the reason for withdrawal, and will provide relevant explanations.

[0092] (1) If the selection of individual subjects violates the inclusion criteria or meets the exclusion criteria, they should not be included in the trial;

[0093] (2) During the trial, adverse events prevented the subject from continuing or completing the trial as scheduled;

[0094] (3) After the subject was given the drug, at T max Vomiting or diarrhea occurred within twice the median time;

[0095] (4) The blood concentration before administration was greater than the corresponding C after administration. max 5%;

[0096] (5) The first sample was C max Furthermore, no subject data was collected from early samples (5-15 minutes after administration);

[0097] (6) During the trial, the subjects did not comply with the trial plan and had poor compliance;

[0098] (7) Data from subjects who did not meet the high-fat, high-calorie requirements after taking a high-fat meal in the postprandial administration trial;

[0099] (8) There are special reasons, including but not limited to violations of the administration protocol, incorrect blood collection time, sample processing, improper storage, or taking drugs that affect the pharmacokinetics of the test drug, which cause a large deviation in the test data.

[0100] The concentration of baloxavir in plasma was determined by HPLC-MS / MS. The validation of the analytical method and the detection of biological samples from subjects should comply with the requirements of Appendix 9012 of the 2020 edition of the Chinese Pharmacopoeia, Volume IV, "Guiding Principles for Validation of Quantitative Analysis Methods for Biological Samples", compiled by the National Pharmacopoeia Commission, and the relevant guiding principles and technical guidelines of the National Medical Products Administration.

[0101] The following pharmacokinetic parameters were determined or calculated using WinNonlin (version 8.2 or later):

[0102] Main parameters: C max AUC 0-72 Secondary parameter: T max , t 1 / 2 , λz .

[0103] Safety evaluation indicators: Observe all subjects for any adverse medical events that occur after administration during the trial, including abnormal clinical symptoms and vital signs, abnormal laboratory test results, and abnormal electrocardiogram results. Record their clinical characteristics, severity, time of occurrence, time of end, duration, treatment measures, and outcomes, and determine their correlation with the study drug.

[0104] Key pharmacokinetic (PK) data point conventions: a) Sampling time points are calculated based on the actual sampling time. b) All blood drug concentrations below the lower limit of quantitation (LOQ) are included in the T1 timeframe for pharmacokinetic analysis. max Previously, values ​​were treated as "0". After Tmax, values ​​were treated as missing. However, for descriptive statistical analysis, values ​​were treated as "0".

[0105] Blood drug concentration (c)-time (t) data analysis: According to the planned blood collection time, individual and mean CT curves and mean semi-log CT curves for the test formulation and the reference formulation were plotted for the subjects; the mean, standard deviation, median, minimum, maximum, lower quartile, upper quartile, geometric mean and coefficient of variation of drug concentration at time points were listed.

[0106] PK parameter analysis: Pharmacokinetic parameters for each subject were calculated using a non-compartmental model. The arithmetic mean, standard deviation, coefficient of variation, median, minimum, maximum, lower quartile, upper quartile, and geometric mean of each parameter were also calculated.

[0107] Analysis of key performance indicators: Using Bioestimated Equation (BES), the key pharmacokinetic parameters (Cmax, AUC) were logarithmically transformed and then subjected to Analysis of Variance (ANOVA). In the ANOVA model, order, drug, and period were treated as fixed effects, while subject (order) was treated as a random effect. The 90% confidence interval of the geometric mean ratio (test formulation / reference formulation) of the key performance indicators was calculated. If it was within the equivalence interval (80.00%~125.00%), it was considered bioequivalent. The results of the two-tailed t-tests were also presented, and a nonparametric test was performed on Tmax.

[0108] Bioequivalence criteria: When the 90% confidence intervals of the geometric mean ratio of the main pharmacokinetic parameters (AUC0-72 and Cmax) of the test formulation and the reference formulation are both within the 80.00% to 125.00% equivalence interval, the two formulations are considered to be bioequivalent.

[0109] Safety analysis: A safety analysis set was used, including adverse events, serious adverse events, changes in clinical laboratory results (complete blood count, blood biochemistry, coagulation parameters, urinalysis, etc.), changes in electrocardiogram, clinical symptoms, and vital signs. Adverse event rates were calculated and systematically categorized. Cross-sectional analysis was performed on laboratory screening and exit test results. Adverse events were coded using the International Medical Terminology Dictionary (MedDRA 26.0 or later) and their incidence rates were grouped and summarized according to the investigational and reference formulations. All adverse events, their severity, and their relevance to the drug were described in tabular form.

[0110] The results are shown in Table 4.

[0111] Table 4 Pharmacokinetic Parameter Results

[0112]

[0113] As shown in Table 4, the absorption rate and extent of the mabaloxavir tablets prepared in this invention are not significantly different from those of the reference preparation in the human body, and they have good bioequivalence.

[0114] In this safety analysis, a total of 100 subjects were included. The results showed that the overall incidence of adverse events (AEs) was 10% (10 / 100), of which 9 were mild, 1 was moderate, and no severe adverse events occurred. The incidence of treatment-related adverse events (TEAEs) was 8% (8 / 100), and the incidence of serious adverse events (SAEs) was 1% (1 / 100). Regarding laboratory tests, compared with the discharge period, the abnormal rate of white blood cell count in complete blood count decreased from 5% to 3%, the abnormal rate of ALT in blood biochemistry decreased from 2% to 1%, the abnormal rate of PT in coagulation parameters increased from 1% to 2%, and the abnormal rate of proteinuria in urinalysis increased from 3% to 4%. In electrocardiogram examination, the abnormal rate of arrhythmia decreased from 2% in the screening period to 1%, and the incidence of QT interval prolongation was 0.5% in both cases. Vital signs examination showed that the incidence of abnormal blood pressure (elevated systolic blood pressure), abnormal heart rate (tachycardia), and abnormal body temperature (fever) was 4%, 3%, and 2% during the screening period, and 3%, 2%, and 1% during the discharge period, respectively. Regarding clinical symptoms, the incidence of headache was 5%, with 70% being mild and 30% being moderate. Overall, among drug-related adverse events (AEs), moderate severity accounted for 20%, the discontinuation rate due to AEs was 0.5% (0.5 / 100), and the average duration of adverse events was 5 days (range 1–14 days). In summary, the investigational formulation demonstrated good safety in the clinical trial, with all indicators within acceptable ranges and no serious safety issues observed.

[0115] Stability test

[0116] High-temperature test: The drug tablets are placed at 60℃, and samples are taken at 10, 20, and 30 days to observe changes in appearance and determine the content of active ingredients, related substances, and other indicators. If there are no significant changes, the drug can be considered stable under high-temperature conditions.

[0117] High humidity test: The drug is placed in a constant temperature and humidity chamber at a relative humidity of 75%. Samples are taken at 10, 20 and 30 days to observe moisture absorption and deliquescence, and to test the drug content, dissolution rate, etc.

[0118] The results are shown in Table 5.

[0119] Table 5 Stability test results

[0120]

[0121] As shown in Table 5, the mabaloxavir tablets prepared in this invention are stable under high temperature and high humidity conditions.

[0122] The mabaloxavir tablets provided by this invention were stored in a light-proof, sealed, cool and dry place (temperature 25℃±2℃, humidity 60%±10%) for long-term testing.

[0123] Samples were taken and tested at the 3rd, 6th, 9th, 12th, 24th, and 36th months. The results showed that the mabaloxavir tablets of this invention exhibited no change in appearance during storage, maintained a content ≥98%, and met the standards for related substances and dissolution. This indicates that the mabaloxavir tablets provided by this invention are stable under long-term conditions and have a shelf life of at least 36 months.

[0124] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for preparing mabaloxavir tablets, characterized in that, The method includes the following steps: The mabaloxavir raw material was completely dissolved in ethanol to obtain a mabaloxavir solution; lactose dodecyl ester and microcrystalline cellulose were mixed and water was added to form a carrier solution; the mabaloxavir solution was slowly added to the carrier solution, stirred evenly, and then sonicated for 10-30 minutes. After removing the solvent by rotary evaporation, microcrystalline mabaloxavir was obtained. Microcrystalline mabaloxavir was mixed evenly with croscarmellose sodium, povidone K25 and stearyl fumarate to obtain a mixture. The mixture was then compressed into tablets to obtain mabaloxavir coated tablets. The amounts of each substance, by weight, are as follows: 15-25 parts mabaloxavir raw material, 5-15 parts lactose dodecyl ester, 5-15 parts croscarmellose sodium, 5-15 parts povidone K25, 20-40 parts microcrystalline cellulose, 0.5-2 parts sodium stearate fumarate, 5-15 parts hydroxypropyl methylcellulose, 0.5-2 parts talc, and 0.5-2 parts titanium dioxide; The preparation method of the lactose dodecyl ester includes the following steps: mixing lactose and dodecyl methyl ester in a molar ratio of 1:1 to 1.5, adding 1% to 5% sulfuric acid of the total molar amount of the reactants, and reacting at 60℃ to 90℃ for 2 to 4 hours to obtain lactose dodecyl ester; The concentration of lactose dodecyl ester in the carrier solution is 5%wt to 15%wt.

2. The preparation method according to claim 1, characterized in that, The purity of the mabaloxavir raw material is ≥99.0%.

3. The preparation method according to claim 1, characterized in that, The ultrasound conditions are 50~100W, 20~40kHz for 15~20 minutes, with each ultrasound session lasting 1~2 minutes and an interval of 30~60 seconds.

4. The preparation method according to claim 1, characterized in that, The conditions for rotary evaporation are 40℃~50℃, 50~150rpm for 1~1.5h.

5. The preparation method according to claim 1, characterized in that, The mabaloxavir tablets are available in 20mg / tablet or 40mg / tablet specifications.

6. The application of the preparation method according to any one of claims 1 to 5 in the preparation of anti-H1N1 drugs.

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