Common tablet containing high-load sargassum pallidum polypeptide and wet preparation method thereof

Abstract

The invention relates to the technical field of pharmaceutical preparations, in particular to a high-load sargassum pallidum polypeptide tablet and a wet preparation method thereof. The tablet comprises the following components: sargassum pallidum polypeptide, lactose 312, microcrystalline cellulose M103, sodium carboxymethyl starch, hydroxypropyl methyl cellulose and magnesium stearate. The preparation method comprises the following steps: mixing the main drug, the filler, the disintegrating agent and the adhesive, carrying out wet granulation, drying until the water content is less than or equal to 3%, finishing granules, adding the lubricant, and tabletting. Through combination of a specific prescription and a wet granulation process, the technical problem that high-load sargassum pallidum polypeptide cannot be directly tableted and formed is successfully solved, and the prepared tablet is moderate in hardness, rapid in disintegration, stable in process and suitable for industrial production.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical preparations, in particular to a common tablet containing high-load Sargassum pallidum polypeptide and a wet preparation method thereof. BACKGROUND

[0002] Sargassum pallidum is a traditional marine medicinal material, which is rich in various bioactive substances. In recent years, Sargassum pallidum polypeptide prepared from it has become a research hotspot due to its small molecular weight and high biological activity, and has shown application potential in the fields of anti-tumor, antioxidant and immune regulation.

[0003] However, its development into stable and effective solid oral preparations faces significant challenges, which restricts its clinical application. In particular, for high drug load tablets, the main difficulties in development exist in the following aspects: first, the physical properties of polypeptide components often have defects. For example, poor flowability and insufficient compressibility, which leads to problems such as cracking and loose tablets when using conventional powder direct compression technology, making it difficult to form. When the polypeptide drug load is high, this problem is particularly prominent, and conventional excipients cannot make up for the lack of formability. Second, to solve the compressibility problem, wet granulation is a commonly used technical means. The selection of the type and amount of binder is crucial in the granulation process. Excessive or improper selection of the binder may improve the compressibility, but it may also excessively prolong the disintegration time of the tablet, even leading to disintegration failure; while insufficient binder cannot improve the compressibility and cannot solve the root problem. Therefore, developing a special wet granulation process suitable for high drug load polypeptide tablets that can simultaneously consider compressibility and good disintegration performance presents a significant technical obstacle. Third, to ensure the content uniformity of high drug load prescriptions, the total mixed granules must have excellent flowability. However, a high proportion of polypeptide drug substances often affects the flow properties of the overall mixture, which raises more precise requirements for the selection and amount of lubricants and flow aids.

[0004] Currently, the development of Sargassum pallidum polypeptide dosage forms is severely lagging behind, especially there is no mature and reliable solution on how to successfully prepare high drug load Sargassum pallidum polypeptide into common tablets that meet the standards of the Chinese Pharmacopoeia and achieve industrial production.

[0005] Therefore, there is an urgent need in the art to develop a tablet formulation and preparation method for high drug load Sargassum pallidum polypeptide to break through the industrialization bottleneck. SUMMARY

[0006] The technical problem solved by the present application is to provide a common tablet containing high-load Sargassum pallidum polypeptide and a wet preparation method thereof to overcome the deficiencies of the prior art.

[0007] To achieve the purpose of the present application, the following technical means are specifically adopted:

[0008] A high-loading sea-buckthorn polypeptide tablet, characterized in that it is composed of the following components in mass percentage:

[0009] sea-buckthorn polypeptide 25-35%,

[0010] disintegrant 0.1-1.5%,

[0011] binder 1.5-4.0%,

[0012] lubricant 0.1-1.0%;

[0013] filler the rest;

[0014] wherein the filler comprises lactose 312 and microcrystalline cellulose M103;

[0015] the tablet is prepared by a wet granulation process.

[0016] The high-loading sea-buckthorn polypeptide tablet, characterized in that the content of sea-buckthorn polypeptide is 31.5%.

[0017] The high-loading sea-buckthorn polypeptide tablet, characterized in that the mass ratio of lactose 312 to microcrystalline cellulose M103 is 1:1 to 4:1.

[0018] The high-loading sea-buckthorn polypeptide tablet, characterized in that the mass ratio of lactose 312 to microcrystalline cellulose M103 is 3:1.

[0019] The high-loading sea-buckthorn polypeptide tablet, characterized in that the disintegrant is sodium carboxymethyl starch, and the content is 0.5%.

[0020] The high-loading sea-buckthorn polypeptide tablet, characterized in that the binder is hydroxypropyl methyl cellulose, and the content is 2.0-3.0%.

[0021] The high-loading sea-buckthorn polypeptide tablet, characterized in that the lubricant is magnesium stearate, and the content is 0.3-0.7%.

[0022] The method for preparing the high-loading sea-buckthorn polypeptide tablet, characterized in that it comprises the following steps:

[0023] (1) uniformly mixing sea-buckthorn polypeptide, disintegrant, binder and filler, wherein the filler comprises lactose 312 and microcrystalline cellulose M103;

[0024] (2) adding 95% ethanol solution to prepare soft material, and sieving and granulating;

[0025] (3) drying at below 50°C to reduce the water content of the granules to 0-3%;

[0026] (4) sieving and sizing, adding lubricant, mixing evenly;

[0027] (5) tabletting, controlling the tablet hardness between 30-60N.

[0028] The high-loading sea-buckthorn polypeptide tablet preparation method is characterized in that the water content of the granules in step (3) is 0-3%.

[0029] The sea-buckthorn polypeptide in the application is a white-like powder, the particle size standard is that 100% can pass through an 80-mesh sieve, the peptide content (based on dry basis) is ≥99%, the molecular weight is ≤1000 Da, the loss on drying is ≤5.0%, the ash content is ≤5.0%, the pesticide residue cannot be detected, the total amount of heavy metals is ≤10 ppm, the arsenic content is ≤10 ppm, the lead content is ≤10 ppm, the total number of colonies is ≤1000 cfu / g, the yeast and mold is ≤100 cfu / g, the E. coli cannot be detected, the Salmonella cannot be detected, and the Staphylococcus cannot be detected.

[0030] Beneficial effects

[0031] Beneficial effects of the application:

[0032] 1. The application surprisingly solves the technical problem that high-loading sea-buckthorn polypeptide cannot be directly tabletted by combining a specific prescription combination with a wet granulation process, and successfully prepares a tablet with good forming.

[0033] 2. The optimized process parameters of the application make the obtained tablet have a disintegration time significantly shorter than 15 minutes specified in the Chinese Pharmacopoeia under the premise of ensuring sufficient hardness, and the process is robust, has good reproducibility, and the product has good stability. DETAILED DESCRIPTION

[0034] The technical solutions of the application will be described below in conjunction with the embodiments of the application. Obviously, the described embodiments are only a part of the embodiments of the application, rather than all the embodiments. The description of the at least one exemplary embodiment is actually only illustrative, but not as any limitation on the application and its application or use. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the application.

[0035] The sea buckthorn polypeptide used in the present application is explained as follows: the manufacturer is Xi'an Muguo Biotechnology Co., Ltd., the product batch number is MG-2024081502, the appearance is white powder, the particle size standard is 100% passing through an 80-mesh sieve, the peptide content (on a dry basis) is 99.07%, the molecular weight is ≤1000 Da, the loss on drying is 0.79%, the ash content is 1.42%, the pesticide residue is not detected, the total amount of heavy metals is ≤10 ppm, the arsenic content is ≤10 ppm, the lead content is ≤10 ppm, the total number of colonies is ≤1000 cfu / g, the yeast and mold is ≤100 cfu / g, the Escherichia coli is not detected, the Salmonella is not detected, and the Staphylococcus is not detected.

[0036] Prescription, preparation and evaluation method of Example 1

[0037] 1. The prescription is initially determined as follows:

[0038] Ingredients Effects Prescription amount ratio (%) Sea buckthorn polypeptide Main drug 31.5％ Lactose, microcrystalline cellulose, corn starch Filling agent 65％ Sodium carboxymethyl starch Disintegrating agent 0.5％ Hydroxypropyl methyl cellulose Binder 2.5％ Magnesium stearate Lubricant 0.5％

[0039] 2. The preparation method is initially determined as follows:

[0040] Each material is passed through an 80-mesh sieve, and the prescription amount of material is weighed. After the main drug, the filler, the disintegrant and the binder in the above prescription are uniformly mixed, 95% ethanol is added, and the material is manually granulated until it is "hand-formed into a group and scattered by light touch". The granules are dried in an oven at 50°C until the water content of the granules is ≤3%, and then the granules are passed through a 20-mesh sieve. The lubricant is added and uniformly mixed to obtain total mixed granules. A 6mm round punch is used, and the target tablet weight is 120mg. The tablets are pressed, and the tablets are obtained.

[0041] 3. The calculation and determination method of part of the evaluation indexes are as follows:

[0042] 3.1 Determination of polypeptide content in the preparation

[0043] The determination method of the sea buckthorn polypeptide in the present application is the biuret method.

[0044] First, the biuret solution is prepared. Copper sulfate 1.59g, potassium sodium tartrate 6.0g and potassium iodide 5.0g are weighed, and water 500ml is added to dissolve. While stirring, 10% sodium hydroxide solution 300ml is added, and water is diluted to 1000ml. After mixing, the biuret solution is obtained.

[0045] Then, the reference solution is prepared. Unless otherwise specified, the bovine serum albumin reference substance is dissolved in water to prepare a solution containing 10mg per 1ml.

[0046] Accurately measure 0.0 ml, 0.2 ml, 0.4 ml, 0.6 ml, 0.8 ml, and 1.0 ml of the reference solution (the volume of the reference solution can be adjusted appropriately within the range of this method), and place them into stoppered test tubes. Add water to each tube to a final volume of 1.0 ml, then add 4.0 ml of biuret reagent to each tube. Mix immediately and incubate at room temperature for 30 minutes. Measure the absorbance at 540 nm using UV-Vis spectrophotometry. Use tube 0 as a blank. Calculate the linear regression equation using the concentration of the reference solution and its corresponding absorbance. Separately measure an appropriate amount of the test solution and perform the same procedure. Calculate the protein concentration in the test solution from the linear regression equation and multiply by the dilution factor to obtain the final concentration.

[0047] 3.2 Angle of repose

[0048] The angle of repose of the particles was measured using the BT-1001 intelligent powder property tester from Dandong Better Instruments Co., Ltd., as a characterization of particle flowability.

[0049] 3.3 Disintegration Time Limit

[0050] According to the 2020 edition of the Chinese Pharmacopoeia, section 0921, the disintegration time test method is used. A lift-type disintegration apparatus is employed. Six tablets of the test sample are taken, and the disintegration apparatus is started for testing. Each tablet should disintegrate completely within 15 minutes. If one tablet fails to disintegrate completely, another six tablets should be taken for retesting, and all of them should meet the requirements.

[0051] Example 2: Investigation of Single-Factor Prescription Composition

[0052] 1. Filler type screening

[0053] Prepare 14 samples according to the formulation in the table below, and control the same tablet hardness range of 30-60N. Screen the filler composition by comparing the preparation process, the angle of repose of the total mixed particles, and the disintegration time.

[0054]

[0055] Based on the above results, products can be successfully prepared using lactose 312 as a filler, or a mixture of lactose 312 and microcrystalline cellulose M101. The total particle flowability is good, and the disintegration time of the prepared tablets meets the requirements of the 2020 edition of the Chinese Pharmacopoeia.

[0056] 2. Lactose type screening

[0057] Lactose 312 is lactose monohydrate, while lactose 310 is anhydrous lactose. Prepare formulations 5-8 according to the table below, controlling the same tablet hardness range of 30-60N. Screen the lactose type by comparing the preparation process, total particle repose angle, and disintegration time.

[0058]

[0059] Results from formulations 5 and 6 show that the product prepared using lactose 312 exhibits excellent total particle flowability, and its disintegration time meets the standards of the 2020 edition of the Chinese Pharmacopoeia. The product prepared using lactose 310 also shows good total particle flowability, but it is lower than that of the lactose 312 group (formulation 5), and its disintegration time does not meet the standards of the 2020 edition of the Chinese Pharmacopoeia. Comparison of data from formulations 7 and 8 further confirms that, compared to lactose 310, the total particle flowability of the mixture prepared using lactose 312 and microcrystalline cellulose M101 is better, and the disintegration time of the compressed tablets is also shorter. In conclusion, lactose 312 is the preferred formulation for this product.

[0060] 3. Screening of microcrystalline cellulose types

[0061] This study compared the differences between M101 and M103, commonly used microcrystalline cellulose grades in tablets. Samples of formulations 9–12 were prepared according to the table below, maintaining the same tableting hardness range of 30–60 N. The microcrystalline cellulose grade was screened by comparing the preparation process, the angle of repose of the total mixed particles, and the disintegration time.

[0062]

[0063] Results from formulations 9 and 10 show that using microcrystalline cellulose M101 alone as a filler resulted in a product with a disintegration time exceeding 15 minutes, failing to meet the standards of the 2020 edition of the Chinese Pharmacopoeia. However, using microcrystalline cellulose M103 alone as a filler resulted in good particle flowability and a product with a disintegration time of less than 15 minutes, meeting the standards of the 2020 edition of the Chinese Pharmacopoeia. Therefore, microcrystalline cellulose M103 is the preferred filler for this product.

[0064] The results from formulations 11 and 12 show that when used in combination with lactose 312 as a filler, the total mixed particles prepared from microcrystalline cellulose M101 (formulation 11) and microcrystalline cellulose M103 (formulation 12) exhibit good flowability, and the disintegration time of the prepared products is significantly improved. This data supports the use of lactose and microcrystalline cellulose as mixed fillers in the formulation of this product.

[0065] 4. Screening of lactose to microcrystalline cellulose ratio

[0066] This study compared the ratios of lactose 312 to microcrystalline cellulose M103 at 1:1, 2:1, 3:1, and 4:1. Samples 13–16 were prepared according to the formulations listed in the table below, with the same tableting hardness range of 30–60 N. The ratio of lactose to microcrystalline cellulose was screened by comparing the preparation process, the angle of repose of the total mixed particles, and the disintegration time.

[0067]

[0068] The results above show that when the ratio of lactose 312 to microcrystalline cellulose M103 in this product is 1:1, 2:1, 3:1, and 4:1, the prepared total mixed particles have good flowability, disintegration time of less than 15 min, and a certain safety range. Therefore, the preferred ratio range of lactose 312 to microcrystalline cellulose M103 is (1-4):1.

[0069] 5. Screening of Lubricant Dosage

[0070] The differences in the amounts of magnesium stearate lubricant in the formulations were compared when they were 0.3%, 0.5%, and 0.7%. Samples 17–19 were prepared according to the formulations in the table below, with the same tablet hardness range of 30–60 N controlled. The amount of lubricant was screened by comparing the preparation process, the angle of repose of the total mixed particles, and the disintegration time.

[0071]

[0072]

[0073] The results above show that the optimal dosage range for the lubricant magnesium stearate in the formulation is 0.3%–0.7%. Sample preparation was smooth, the total mixed particles exhibited good flowability, and the disintegration times of the compressed tablets all met the standards of the 2020 edition of the Chinese Pharmacopoeia. Therefore, the preferred dosage range for the lubricant magnesium stearate is 0.3%–0.7%.

[0074] 6. Adhesive dosage screening

[0075] The differences in the amount of hydroxypropyl methylcellulose binder in the formulations were compared when they were 2%, 2.5%, and 3%. Samples 20–22 were prepared according to the formulations listed in the table below, maintaining the same tableting hardness range of 30–60 N. The amount of binder was screened by comparing the preparation process, the angle of repose of the total mixed particles, and the disintegration time.

[0076]

[0077] The results above show that the optimal range for the amount of hydroxypropyl methylcellulose (HPMC) binder in the formulation is 2.0%–3.0%. Sample preparation was smooth, the total mixed particles exhibited good flowability, and the disintegration times of the compressed tablets all met the standards of the 2020 edition of the Chinese Pharmacopoeia. Therefore, the preferred range for the amount of HPMC binder is 2.0%–3.0%.

[0078] Example 3: Screening of Preparation Process

[0079] 1. Screening of tablet hardness range

[0080] The differences in tablet hardness ranges of 20-30N, 30-60N, and 60-90N were compared. Samples 23-25 ​​of the formulation were prepared according to the table below, and the appropriate tablet hardness range was screened by comparing the preparation process and disintegration time.

[0081]

[0082] The data above indicates that the preferred tablet hardness range is 30-60N.

[0083] 2. Screening of moisture content range for particle drying

[0084] Compare the differences in the dry moisture content ranges of 0%-3% and 3%-6%. Prepare samples 26-27 according to the composition of the following table, and screen the appropriate dry moisture content range of the particles by comparing the preparation process, the flowability of the total mixed particles, and the disintegration time.

[0085]

[0086]

[0087] The data above shows that the preferred moisture content range for drying particles is 0%-3%.

[0088] Example 4 Quality Evaluation

[0089] Formula 26 was used, with a batch size of 10,000 tablets. After tablet compression, samples were taken for the following tests.

[0090] 1. Appearance

[0091] This product is a light yellow, round tablet with a smooth surface and uniform color.

[0092] 2. Tablet weight difference

[0093] According to the 2020 edition of the Chinese Pharmacopoeia, 20 tablets of Artemisia annua polypeptide were taken, and the total weight was accurately weighed. After obtaining the average tablet weight, the weight of each tablet was accurately weighed. The weight of each tablet was compared with the average tablet weight. According to the table, no more than 2 tablets exceeded the weight difference limit, and no tablet exceeded the limit by more than 1 time.

[0094] Average tablet mass or labeled tablet mass / g Mass difference limit / % 0.30 g or less ±7.5％ 0.30 g and 0.30 g or more ±5％

[0095] Following the above method, 20 tablets were weighed as follows (unit: mg): 129.1, 116.3, 119.5, 121.8, 112.0, 124.2, 120.6, 117.9, 125.2, 114.3, 122.0, 123.5, 116.7, 123.1, 126.5, 119.8, 115.3, 128.9, 121.1, 113.3. Based on the above criteria, the tablet weight variation of this product is acceptable.

[0096] 3. Disintegration time limit

[0097] The disintegration time limit test method is determined according to the 2020 edition of the Chinese Pharmacopoeia, 0921. Take 6 tablets of the test sample and place them in the glass tubes of the above-mentioned basket. Start the disintegration tester and check that each tablet disintegrates completely within 15 minutes.

[0098] According to the above regulations, 6 tablets were randomly sampled, and the disintegration times were 9 min 32 s, 10 min 18 s, 8 min 51 s, 9 min 27 s, 9 min 03 s, and 9 min 21 s, which meet the requirements of the Chinese Pharmacopoeia and are qualified for the disintegration time limit.

[0099] 4. Hardness

[0100] Six pieces of this product were randomly selected, and the hardness was measured as follows: 42.6N, 37.1N, 35.5N, 50.0N, 41.8N, and 39.1N.

[0101] 5. Friability

[0102] The tablet friability test should be performed according to the 2020 edition of the Chinese Pharmacopoeia. For tablets weighing 0.65g or less, take a number of tablets to bring the total weight to approximately 6.5g. Blow away any loose powder using a blower, weigh accurately, place in a cylinder, and rotate 100 times. Remove, remove powder using the same method, weigh accurately, and the weight loss should not exceed 1%, and no broken, cracked, or pulverized tablets should be detected.

[0103] Results: The weight loss of this product was 0.2%, and no broken, cracked, or pulverized pieces were detected, indicating that the friability was acceptable.

[0104] 6. Stability Test

[0105] The prepared samples were sealed and packaged, and placed at 40℃±2℃ and relative humidity 75%±5% for 6 months to examine the quality of the products. The results are as follows:

[0106] Time (month) Appearance Moisture (%) Disintegration time Hardness (N) Polypeptide content in tablets determined by biuret method (%) 0 Smooth and complete 1.69 9 min 51 s 40.5 6.70 1 Smooth and complete 1.72 9 min 32 s 39.7 6.75 2 Smooth and complete 1.81 10 min 21 s 42.8 6.63 3 Smooth and complete 1.78 10 min 37 s 41.6 6.79 6 Smooth and complete 1.85 10 min 56 s 42.3 6.66

[0107] Note: In the biuret method experiment used in this invention, the reference standard is bovine serum albumin. Therefore, the polypeptide content in the seaweed polypeptide tablets is calculated based on bovine serum albumin, which differs from the theoretical content of seaweed polypeptide.

[0108] Comparative Example 1: Direct compression of powder into tablets

[0109] Prepare samples according to prescriptions 26-30 in the table below. Select the direct powder compression method to prepare samples. The specific method is as follows: all materials are passed through an 80-mesh sieve. Weigh the prescribed amount of materials. Mix the main drug, filler, disintegrant, binder and lubricant in the above prescriptions evenly. Use a 6mm round die to compress the tablets. The target tablet weight is 120mg.

[0110]

[0111] The results above show that, using the same formulation as this invention (Formulation 26), direct powder compression is completely impossible. This demonstrates that the wet granulation process used in this invention is not obvious but crucial for this specific high-loading formulation.

[0112] Comparative Example 2: Comparison of Different Adhesives

[0113] Prepare samples according to prescriptions 26-34 in the table below and compare the effects of different adhesives.

[0114]

[0115] The results above show that when hydroxypropyl methylcellulose is used as the binder, the disintegration time of the compressed tablets is less than 15 minutes.

[0116] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A high-load Artemisia annua polypeptide tablet, characterized in that, It consists of the following components by mass percentage: Artemisia annua polypeptide 25-35%, Disintegrant 0.1-1.5%, Adhesive 1.5-4.0%, Lubricant 0.1-1.0%; Filler allowance; The filler contains lactose 312 and microcrystalline cellulose M103; The tablets are prepared using a wet granulation process.

2. The high-load Artemisia annua polypeptide tablet according to claim 1, characterized in that, The content of the seaweed polypeptide is 31.5%.

3. The high-load Artemisia annua polypeptide tablet according to claim 1 or 2, characterized in that, The mass ratio of lactose 312 to microcrystalline cellulose M103 is 1:1 to 4:

1.

4. The high-load Artemisia annua polypeptide tablet according to claim 3, characterized in that, The mass ratio of lactose 312 to microcrystalline cellulose M103 is 3:

1.

5. The high-load Artemisia annua polypeptide tablet according to claim 1, characterized in that, The disintegrant is sodium carboxymethyl starch, with a content of 0.5%.

6. The high-load Artemisia annua polypeptide tablet according to claim 1, characterized in that, The adhesive is hydroxypropyl methylcellulose, with a content of 2.0-3.0%.

7. The high-load Artemisia annua polypeptide tablet according to claim 1, characterized in that, The lubricant is magnesium stearate, with a content of 0.3-0.7%.

8. A method for preparing high-load Artemisia annua polypeptide tablets as described in any one of claims 1-7, characterized in that, Includes the following steps: (1) The seaweed polypeptide, disintegrant, binder and filler are mixed evenly, wherein the filler contains lactose 312 and microcrystalline cellulose M103; (2) Add 95% ethanol solution to prepare a soft mass, then sieve and granulate; (3) Dry at a temperature below 50°C to reduce the moisture content of the particles to 0-3%; (4) Sieve and granulate, add lubricant, and mix evenly; (5) Compress the tablets and control the tablet hardness between 30-60N.

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