JP7740652B2 - Enteric-coated solid dosage forms - Google Patents

Description

The present invention relates to an enteric-coated solid preparation.

In recent years, commercially available enteric-coated solid formulations commonly contain lactoferrin and lactic acid bacteria as active ingredients. Lactoferrin is known to exhibit various physiological functions, including improving bowel movements, improving intestinal flora, and maintaining immunity, and a variety of products containing it, including functional food and health foods, are already on the market. Furthermore, lactic acid bacteria (e.g., Lacticaseibacillus casei) are known to exhibit physiological functions such as improving bowel movements, as well as preventing and alleviating allergic symptoms such as atopic dermatitis and hay fever. Because the shared physiological functions of lactoferrin and lactic acid bacteria often have different mechanisms of action, combining the two ingredients can simultaneously exhibit the desired physiological functions and synergistically promote health. Furthermore, because lactoferrin and lactic acid bacteria are easily decomposed by gastric acid and digestive enzymes such as pepsin, enteric-coated solid preparations that do not dissolve in the stomach but dissolve in the intestines have been developed to maximize their physiological functions.

In order to achieve excellent intestinal environment improvement effects, an intestinal environment improvement agent containing lactoferrin and killed Lactobacillus brevis bacteria as active ingredients has been proposed (see, for example, Patent Document 1). However, the intestinal environment improvement agent in Patent Document 1 suffers from the problem that, when stored in a high-temperature environment, it changes over time, reducing the lactoferrin's ability to dissolve in intestinal fluid. Furthermore, when lactoferrin is used in combination with lactic acid bacteria coated with peptidoglycan, an insoluble polymer, the lactoferrin's ability to dissolve in intestinal fluid over time is further reduced.

Furthermore, the intestinal environment improving agent in Patent Document 1 had problems with gastric juice resistance, such as elution of the active ingredient from pores in the enteric coating in acidic solutions such as gastric juice, and holes due to non-uniformity in the enteric coating leading to formulation disintegration. These problems are thought to be exacerbated by deterioration of the enteric coating components when stored in a high-temperature environment.

Therefore, an enteric-coated solid dosage form that has excellent gastric juice resistance and high intestinal juice dissolution even after aging has not yet been provided, and there is currently a strong demand for its prompt development.

JP 2014-111668 A

The present invention aims to solve the above-mentioned problems of the past and achieve the following objectives: Namely, the objective is to provide an enteric-coated solid preparation that has excellent resistance to gastric juice and high intestinal juice dissolution even after aging.

As a result of extensive research to achieve the above-mentioned object, the inventors have discovered that the enteric coated solid formulation of the present invention, which contains (A) lactoferrin, (B) lactic acid bacteria, and (C) dextrin having a dextrose equivalent (DE) of 30 or less, has excellent resistance to gastric juice and high intestinal juice dissolution even after aging.

The present invention is based on the above findings of the present inventors, and the means for solving the above problems are as follows.
<1> An enteric coated solid preparation containing (A) lactoferrin, (B) lactic acid bacteria, and (C) dextrin having a dextrose equivalent (DE) of 30 or less.
<2> The enteric coated solid preparation according to <1>, wherein the dextrose equivalent (DE) of the (C) component is 10 or more and 30 or less.
<3> The enteric coated solid preparation according to <1>, wherein the mass ratio of the content of the component (C) to the content of the component (A) [(C)/(A)] is 0.05 or more and 0.6 or less.
<4> The enteric coated solid preparation according to any one of <1> to <3>, wherein the component (B) contains 100 million to 300 billion bacterial cells per tablet.
<5> The enteric coated solid preparation according to any one of <1> to <4>, wherein the excipient comprises at least one selected from cellulose or a cellulose derivative, a sugar, or a sugar alcohol.

The present invention solves the above-mentioned problems of the prior art and achieves the above-mentioned objectives, thereby providing an enteric-coated solid preparation that has excellent gastric juice resistance and high intestinal juice dissolution even after aging.

(Enteric coated solid preparations)
The enteric coated solid preparation of the present invention contains (A) lactoferrin, (B) lactic acid bacteria, and (C) dextrin having a dextrose equivalent (DE) of 30 or less, and may further contain other ingredients as necessary.
In this specification, (A) lactoferrin may be referred to as "(A) component," (B) lactic acid bacteria may be referred to as "(B) component," and (C) dextrin with a dextrose equivalent (DE) of 30 or less may be referred to as "(C) component" or "(C) dextrin."

<(A) Lactoferrin>
The (A) lactoferrin is contained to exhibit physiological functions such as a function to improve lipid metabolism, a function to improve constipation, a function to improve intestinal flora, a function to improve sleep, a function to promote secretion of growth hormone, a function to improve dry eye and corneal epithelial abrasion, a function to improve eye disorders caused by dry eye and corneal epithelial abrasion, a function to lower blood pressure, a function to improve allergies, a function to maintain immune function, a function to improve immune function, a function to improve immune function, a function to improve liver function, etc. In particular, as a function to improve lipid metabolism, it is useful for reducing visceral fat and BMI, and for exhibiting the functions of maintaining, improving, and improving immune function.

The lactoferrin (A) is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include lactoferrin isolated from mammalian colostrum, transitional milk, normal milk, terminal milk, etc., lactoferrin isolated from processed mammalian milk products such as skim milk and whey, lactoferrin produced from plants, lactoferrin obtained by genetic recombination, etc. Among these, lactoferrin concentrates containing 50% by mass or more of lactoferrin monomer are preferred.
The mammal is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include humans, cows, sheep, goats, and horses.
The plant is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include rice.

The method for separating lactoferrin is not particularly limited and can be appropriately selected depending on the purpose. For example, ion exchange chromatography can be used.
The lactoferrin to be used may be a commercially available product, or may be prepared by a known method.

From the viewpoint of the expression of physiological functions, the content of (A) lactoferrin in the enteric solid preparation is preferably 1% by mass or more and 60% by mass or less, more preferably 4% by mass or more and 60% by mass or less, even more preferably 15% by mass or more and 56% by mass or less, and particularly preferably 18% by mass or more and 50% by mass or less.
When the content of (A) lactoferrin in the enteric coated solid preparation is 1% by mass or more, physiological functions are exhibited, tableting problems (binding) are suppressed, the number of tablets to be taken can be kept within an appropriate range, and ease of administration is improved, which is preferable.
When the content of the (A) lactoferrin in the enteric coated solid preparation is 60% by mass or less, the initial dissolution rate and disintegration rate of the lactoferrin, the retention rate of dissolution rate of lactoferrin, the retention rate of disintegration rate, the resistance to gastric juice after aging, and the moldability of the uncoated tablet are improved, which is preferable. Note that the "retention rate of disintegration rate" in the present invention indicates the degree to which the disintegration rate of the tablet after aging is maintained compared to that of the tablet immediately after production.

There are no particular restrictions on the daily intake of (A) lactoferrin and it can be selected appropriately depending on the purpose. However, from the viewpoint of the expression of physiological functions, it is preferably 50 mg to 1000 mg, more preferably 100 mg to 500 mg, and even more preferably 200 mg to 400 mg.

The method for producing the lactoferrin is not particularly limited, and lactoferrin produced by a conventional method can be used, which may include a drying step, for example.
Examples of drying steps in the lactoferrin production method include freeze-drying and spray-drying. The lactoferrin obtained by the drying step is preferably contained in the dried product at a purity of 85% or more. Furthermore, the loss on drying in the drying step is preferably 6% by mass or less.
The method for producing lactoferrin may further include processing steps such as granulation, particle coating, and pulverization.

The average particle size of the lactoferrin (A) is not particularly limited and can be appropriately selected depending on the purpose, but is preferably 20 μm or more and 300 μm or less, and more preferably 20 μm or more and 100 μm or less.
In the present invention, the average particle size refers to the 50% diameter (median diameter, volume basis) in the particle size distribution by laser diffraction scattering.

<(B) Lactic acid bacteria>
The (B) lactic acid bacteria are contained to exhibit functions such as improving bowel movements, improving intestinal flora, maintaining skin moisture, protecting skin from dryness, maintaining skin barrier function, improving skin texture, preventing rough skin, maintaining skin flexibility, protecting skin from ultraviolet rays, giving skin firmness, giving skin luster, smoothing skin, improving sleep quality, improving immunity, preventing allergic symptoms such as atopic dermatitis and hay fever, alleviating allergic symptoms, improving blood flow, lowering blood pressure, preventing periodontal disease, and preventing bad breath. In particular, ingestion of the (B) lactic acid bacteria prevents moisture loss from the skin, and is therefore useful for exhibiting the functions of protecting skin from dryness, maintaining skin barrier function, and improving bowel movements.

The lactic acid bacteria (B) are not particularly limited and can be appropriately selected depending on the purpose. Examples include lactic acid bacteria belonging to the genus Lactobacillus, Lacticaseibacillus, Lactiprantibacillus, Lactilactobacillus, Rigilactobacillus, Leviractobacillus, Rimosilactobacillus, Enterococcus, Lactococcus, Streptococcus, Staphylococcus, and Bifidobacterium.
Examples of the lactic acid bacteria belonging to the genus Lactobacillus include Lactobacillus gasseri, Lactobacillus paragasseri, Lactobacillus crispatus, and Lactobacillus helveticus.
Examples of the lactic acid bacteria belonging to the genus Lacticaseibacillus include Lacticaseibacillus casei, Lacticaseibacillus paracasei, Lacticaseibacillus rhamnosus, and Lacticaseibacillus curvatus.
Examples of the lactic acid bacteria belonging to the genus Lactiplantibacillus include Lactiplantibacillus plantarum and Lactiplantibacillus pentosus.
Examples of the lactic acid bacteria belonging to the genus Lactilactobacillus include Lactilactobacillus sakei.
Examples of the lactic acid bacteria belonging to the genus Ligilactobacillus include Ligilactobacillus salivarius.
Examples of the lactic acid bacteria belonging to the genus Levilactobacillus include Levilactobacillus brevis.
Examples of the lactic acid bacteria belonging to the genus Limosilactobacillus include Limosilactobacillus reuteri.
Examples of the lactic acid bacteria belonging to the genus Enterococcus include Enterococcus faecalis and Enterococcus faecium.
Examples of the lactic acid bacteria belonging to the genus Lactococcus include Lactococcus lactis and Lactococcus cremoris.
Examples of the lactic acid bacteria belonging to the genus Streptococcus include Streptococcus thermophilus.
Examples of the lactic acid bacteria belonging to the genus Staphylococcus include Staphylococcus xylosus and Staphylococcus carnosus.
Examples of the lactic acid bacteria belonging to the genus Bifidobacterium include Bifidobacterium animalis lactis, Bifidobacterium animalis longum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium adolescentis, and Bifidobacterium infantis. The lactic acid bacteria of the genus Bifidobacterium are sometimes called bifidobacteria.
In 2020, Zheng et al. reclassified the Lactobacillus genus into a total of 25 genera. For example, the classification name of Lactobacillus casei was changed to Lacticaseibacillus casei. (Int. J. Sys. Evol. Microbiol., 2020; 70: 2782-2858)
These may be used alone or in combination of two or more.

The (B) lactic acid bacteria are not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include those derived from the human intestinal tract, dairy products, and plants (plant-derived products including processed foods such as miso, soy sauce, pickles, bran, grass, rice, wheat, and malt). Among these, those isolated from rice or processed rice foods are preferred.
The (B) lactic acid bacteria may be viable cells or sterilized cells that have been subjected to a sterilization process.
The (B) lactic acid bacteria may be a dispersion with an additive such as an excipient, which will be described later. Details of the case where the (B) lactic acid bacteria is a dispersion with an additive will be described later.

The content of the (B) lactic acid bacteria in the enteric coated solid preparation of the present invention is not particularly limited and can be appropriately selected depending on the dosage form, administration mode, and recipient. However, the content is preferably such that 1 billion to 1 trillion bacteria can be ingested per day by an adult, more preferably 2 billion to 500 billion bacteria, and even more preferably 5 billion to 100 billion bacteria can be ingested per day by an adult.
It is preferable to contain the lactic acid bacteria (B) so that one adult can ingest 1 billion or more per day, as this will allow physiological functions to be expressed and resistance to gastric juices to be improved over time.
The content of the (B) lactic acid bacteria is preferably such that one adult can ingest 1 trillion or less per day, since this improves the initial disintegration property, disintegration property over time, initial dissolution rate, dissolution retention rate over time, and moldability of the preparation.

The number of cells of the (B) lactic acid bacteria per tablet is preferably 100 million to 300 billion, more preferably 500 million to 160 billion, and particularly preferably 1 billion to 100 billion.
The number of cells of the lactic acid bacteria (B) contained per tablet is preferably 100 million or more, as this allows physiological functions to be expressed and improves gastric juice resistance over time.
The number of cells of the (B) lactic acid bacteria per tablet is preferably 300 billion or less, since this improves the initial disintegration property and disintegration property over time, the initial dissolution rate, the maintenance rate of dissolution over time, and the moldability of the formulation.

The content of the (B) lactic acid bacteria in the enteric solid preparation is preferably 0.05% by mass or more and 35% by mass or less, more preferably 0.1% by mass or more and 30% by mass or less, even more preferably 0.2% by mass or more and 25% by mass or less, and particularly preferably 3% by mass or more and 20% by mass or less.
When the content of the (B) lactic acid bacteria in the enteric coated solid preparation is 0.05% by mass or more, physiological functions are exhibited and resistance to gastric juice is improved, which is preferable.
When the content of the (B) lactic acid bacteria in the enteric coated solid preparation is 35% by mass or less, the initial disintegration property, disintegration property over time, initial dissolution rate, dissolution retention rate over time, and moldability of the preparation are improved, which is preferable. Note that when the (B) component is a triturated product containing additives such as excipients described later, the content of the (B) lactic acid bacteria does not include the additives.

There are no particular limitations on the method for measuring the number of lactic acid bacteria contained in the raw material powder or solid preparation, and it can be selected appropriately depending on the purpose. Examples include hemocytometer measurement and DAPI staining. Of these, measuring the number of bacteria using DAPI staining is preferred.

Specific examples of measuring the number of bacteria by the DAPI staining method include the following method.
0.1 g of sample was weighed into a 15 ml centrifuge tube and diluted 100-fold with phosphate-buffered saline (34 g of anhydrous potassium dihydrogen phosphate was dissolved in purified water, and 1 M sodium hydroxide was added to adjust the pH to 7.2, bringing the total volume to 1000 mL). The mixture was then mixed. The mixture was homogenized using a vortex mixer and ultrasonic treatment, resulting in a stock sample solution. The stock sample solution was then diluted appropriately to prepare a measurement sample solution.
The sample solution for measurement was filtered through a membrane filter with a pore size of 0.20 μm, then reacted with a DAPI solution (reagent) for about 1 minute, and filtered through a filtration device. The cells captured on the membrane filter were photographed using an epifluorescence microscope. The number of cells in the photographed image and the average number of cells per image were counted, and the total number of bacteria per gram of sample was calculated using the following formula:
Total number of bacteria (cells/g) = average number of cells per image x effective filtration area / area of one image / filtration amount Note that the effective filtration area should be determined by checking the specifications of the filtration device to be used.

The (B) lactic acid bacteria may be commercially available products or those prepared by known methods.
The method for preparing the (B) lactic acid bacteria is not particularly limited and can be appropriately selected depending on the purpose, and can be obtained, for example, by freeze-drying a bacterial cell liquid containing live lactic acid bacteria. Furthermore, before freeze-drying, additives such as excipients, dispersants, and protective agents may be added as necessary to obtain a lactic acid bacteria dilution product.
In the method for preparing lactic acid bacteria (B) above, when sterilized lactic acid bacteria are used as a raw material, a bacterial cell liquid containing live lactic acid bacteria may be sterilized by, for example, a heat method such as moist heat sterilization, dry heat sterilization, or high-frequency sterilization; a gas method such as ethylene oxide gas sterilization or hydrogen peroxide sterilization; or a radiation method such as gamma ray irradiation sterilization or electron beam irradiation sterilization.
The (B) lactic acid bacteria may be stored frozen.

The additives are not particularly limited and can be selected appropriately depending on the purpose. Examples include sugars such as dextrin, trehalose, potato starch, sucrose, and oligosaccharides; proteins such as milk protein; and lipids. When dextrin is used as the additive, it is preferable to use dextrin with a dextrose equivalent (DE) of 30 or less, as described below. When dextrin is used as the additive, the additive may be the entire amount of dextrin contained in the enteric-coated solid formulation of the present invention, or it may be a combination of dextrin and other optional ingredients.

When the (B) lactic acid bacteria are dispersed with the additive, the content of the (B) lactic acid bacteria in the dispersion is not particularly limited and can be selected appropriately depending on the purpose, but it is preferably contained in an amount of 1% by mass or more and 90% by mass or less, and more preferably contained in an amount of 10% by mass or more and 60% by mass or less.
The content of the (B) lactic acid bacteria in the diluted product is preferably 1% by mass or more, since the physiological functions are easily expressed. The content of the (B) lactic acid bacteria in the diluted product is preferably 90% by mass or less, so that sufficient hardness and good disintegrability are obtained.

The lactic acid bacteria used to prepare the (B) lactic acid bacteria can be obtained, for example, from organizations such as ATCC (registered trademark) or IFO, the Japan Bifidobacteria Center, or the Patent Microorganisms Deposit Center of the National Institute of Technology and Evaluation.

<(C) Dextrin>
The dextrin of the component (C) is contained as an excipient, binder, etc. in the solid preparation, and in the present invention, improves the retention rate of dissolution in intestinal fluid and resistance to gastric juice.
Dextrins are obtained by hydrolyzing starch and are classified according to their dextrose equivalent (saccharification rate). The dextrose equivalent (saccharification rate) can be expressed as a DE value. For example, a DE value closer to 0 indicates that the dextrose has properties closer to those of starch, and a DE value closer to 100 indicates that the starch has been hydrolyzed to a greater extent.
In the present invention, the "maintenance rate of intestinal fluid dissolution" indicates the degree to which the intestinal fluid dissolution property of a tablet is maintained after aging compared to that of the tablet immediately after production.

The dextrin in the enteric coated solid preparation of the present invention has a dextrose equivalent (DE value) of 30 or less.
The dextrose equivalent (DE value) of the component (C) is preferably 10 or more and 30 or less, and more preferably 10 or more and 20 or less.
If the dextrose equivalent (DE value) of the component (C) is 10 or more, the retention rate of intestinal fluid elution is improved, which is preferable.
If the dextrose equivalent (DE value) of the component (C) is 30 or less, the retention rate of intestinal fluid elution and resistance to gastric juice over time are improved, which is preferable.
In addition, as long as the final DE value measured in the enteric coated solid preparation of the present invention is 30 or less, a plurality of dextrins having different DE values may be used in combination.

The method for measuring the dextrose equivalent (DE value) of the component (C) is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include the following methods (I) to (III). Among these, measurement by the measurement method (II) is preferred.
The DE value is not limited to dextrin alone, but can also be measured by the same method for dextrin in a preparation mixed with an excipient or a functional ingredient, and for dextrin in a lactic acid bacteria raw material produced by adding dextrin as a dispersion medium. However, when modified milk powder is contained in the preparation, it is preferable to measure the DE value by the measurement method (III).
(I) Consumer Affairs Agency, "Specifications and Standards for the Specified Health Foods (Standards and Standards Type) System," 2014. The measurement method for indigestible dextrin in the "(Appendix) Component Standards" (measurement temperature: 15-30°C)
(II) The measurement method described in the Central Customs Analysis Laboratory, "Quantitative Analysis Method for Reducing Sugars in Dextrin" (measurement temperature: 15-30°C)
(III) The measurement method described in the Central Customs Analysis Laboratory Report (33), "Measurement of DE of Dextrin in Formulated Powdered Milk," Central Customs Analysis Laboratory, 1994, pp. 23-28 (measurement temperature: 15-30°C).

A specific method for measuring (II) above is as follows.
(II-1) The analytical sample to be analyzed is prepared as follows, depending on the properties of the sample.
If the sample to be analyzed is a solid, it should be in powder or crystalline form, and any lumps present should be crushed and mixed thoroughly. If the sample to be analyzed is a liquid, and crystals or lumps have precipitated in the liquid, place it in a sealed container, immerse it in a water bath at 60-70°C to dissolve, shake well, mix, and then cool to room temperature.
(II-2) The moisture content M (%) is determined.
If the sample prepared in (II-1) above is a solid sample, accurately weigh out about 2 g into a weighing bottle adjusted to a constant weight in advance and dry in a vacuum dryer for 4 hours at 70°C. Next, allow to cool to room temperature in a desiccator, then weigh it, and repeat vacuum drying for 1 hour at a time. When the weight loss is 2 mg or less, it is considered to have reached a constant weight.
If the sample prepared in (II-1) above is a liquid sample, approximately 15 g of sea sand (or 5 g of Celite) is placed in a weighing bottle as a drying aid. This is then dried in a 105°C oven along with a glass rod large enough to fit in the bottle, and a constant weight is determined. Next, an amount equivalent to approximately 2 g of solids is accurately weighed out of the homogenous sample prepared in (II-1) above. If necessary, a small amount of water is added until the entire sample is submerged. Heat the sample in a water bath while occasionally stirring with a glass rod to volatilize most of the water. The sample is then dried in a 105°C oven with occasional stirring until nearly dry, after which it is transferred to a vacuum oven and dried at 70°C for 4 hours. Allow the sample to cool to room temperature in a desiccator and then weigh the sample. Repeat vacuum drying for one hour at a time; a constant weight is considered to have been reached when the weight loss is 2 mg or less.
When W ₀ is the amount of sample (g) and W ₀ is the weight (g) of the sample after drying, the moisture content M (%) can be calculated by the following formula (1).
Water content M (%)={(W ₀ −W ₁ )/W ₀ }×100 (1) Formula (II-3) Fehling's solution is prepared.
34.639 g of copper sulfate ( _CuSO4.5H2O ) was dissolved in water to make 500 mL, and _the solution was left to stand for two days and then filtered to prepare Fehling's solution A. Furthermore, 173 g of _potassium sodium tartrate ( _{KNaC4H4O6.4H2O} ) and ₅₀ g _of sodium hydroxide were dissolved in water to make 500 mL, and the solution was left to stand for two days and then filtered to prepare Fehling's solution B.
5.0 mL of Fehling's solution A and 5 mL of Fehling's solution B are placed in a 200 mL Erlenmeyer flask, and 19.5 mL of standard invert sugar solution is added using a 50 mL burette. After boiling for 2 minutes on an electric heater, 4 drops of methylene blue solution are added, and while boiling, the standard invert sugar solution is added dropwise. The endpoint is when the blue color disappears. The titration is completed within 3 minutes after the start of boiling. If A is the amount (mL) of standard invert sugar solution consumed, the titer of Fehling's solution can be calculated using the following formula (2).
Titer = 20.36/A (2) The standard invert sugar solution was prepared by accurately weighing 4.75 g of sucrose (reagent) and transferring it to a 500 mL measuring flask using 90 mL of water, adding 5 mL of hydrochloric acid (specific gravity 1.18), and leaving it at 20 to 30°C for 3 days, then adding water to the constant volume. This was stored in a cool, dark place, and then 50 mL was transferred to a 200 mL measuring flask, neutralized with 1 mol/L sodium hydroxide solution using phenolphthalein as an indicator, and added water to the constant volume.
(II-4) Quantifying the reducing sugar content.
About 10 g of the sample prepared in (II-1) above was accurately weighed, dissolved in water, transferred to a 500 mL volumetric flask, and water was added to the flask to make the volume constant, which was used as the test solution.
5.0 mL of Fehling's solution A and 5 mL of Fehling's solution B were placed in a 200 mL Erlenmeyer flask, and 15 mL of test solution was added using a 50 mL burette. After boiling for 2 minutes on an electric heater, 4 drops of methylene blue solution were added, and the test solution was added dropwise while boiling. The endpoint was the disappearance of the blue color. The titration was completed within 3 minutes after boiling began, and this was considered a preliminary titration. An additional amount of test solution approximately 1 mL less than the titration constant obtained in the preliminary titration was added, and titration was repeated in the same manner. The amount of test solution consumed was multiplied by the titer of Fehling's solution, and the reducing sugar concentration (mg/100 mg) was calculated as glucose using the rhein-enoin sugar table (glucose) shown in Table 1 below.

(II-5) The reducing sugar content of a sample in a dry state, calculated as glucose, can be calculated using the following formula (3): DE (%) is the reducing sugar content (%) of the sample in a dry state, calculated as glucose, Ds is the amount of glucose (mg) in 100 mL of test solution obtained using Table 1, M is the moisture content (%) of the sample, and S is the amount of sample collected (g).
DE (%) = [Ds/{2(100-M)S}]×100...Equation (3)

When measuring the DE value of dextrin in a lactic acid bacteria raw material produced by adding dextrin as a dispersion medium, it is preferable to dissolve the sample in the above-mentioned (II-4), filter the sample through filter paper in advance to remove impurities, and then filter the sample again through a membrane filter of 1 μm or less to remove insoluble matter such as lactic acid bacteria, and then perform the measurement on the test solution.
When the DE value is not measured for dextrin alone, it can be calculated using the following formula (4), which takes into account the mass ratio of dextrin in the sample, instead of formula (3) in (II-5) above, where R is the mass ratio (%) of dextrin in the sample.
DE (%) = [Ds/{2(100-M)S×R/100}]×100...Equation (4)

The content of the (C) dextrin in the enteric coated solid preparation is preferably 0.05% by mass or more and 35% by mass or less, more preferably 0.2% by mass or more and 30% by mass or less, even more preferably 0.75% by mass or more and 19% by mass or less, and particularly preferably 0.9% by mass or more and 15% by mass or less.
When the content of the dextrin (C) in the enteric coated solid preparation is 0.05% by mass or more, the maintenance rate of intestinal fluid dissolution is improved, and the moldability is also improved, which is preferable.
When the content of the dextrin (C) in the enteric coated solid preparation is 35% by mass or less, resistance to gastric juice after aging is improved, adhesion of the tablets during tableting is suppressed, and tableting problems such as binding are suppressed, which is preferable.

<Mass ratio [(C)/(A)]>
In the present invention, the mass ratio of the content of the component (C) to the content of the component (A) [(C)/(A)] is preferably 0.05 or more, more preferably 0.05 or more and 0.6 or less, and even more preferably 0.1 or more and 0.36 or less, from the viewpoints of intestinal fluid dissolution property and gastric juice resistance.
When the mass ratio [(C)/(A)] is 0.05 or more, the retention rate of intestinal fluid elution is improved, and moldability is also improved, which is preferable.
When the mass ratio [(C)/(A)] is 0.6 or less, resistance to gastric juice over time is improved, adhesion to disks during tableting is suppressed, and tableting problems such as binding are suppressed, which is preferable.

<Other ingredients>
Examples of the other components include excipients other than component (C), disintegrants, fluidizing agents, lubricants, binders, coating agents, enteric coating agents, and the like.
The other components may be used alone or in combination of two or more.

- (C) Excipients other than the component -
The excipient other than the component (C) (hereinafter, sometimes referred to as "excipient") is not particularly limited and can be appropriately selected depending on the purpose. Examples thereof include cellulose or cellulose derivatives, sugar alcohols, sugars (monosaccharides, disaccharides, oligosaccharides, polysaccharides), calcium hydrogen phosphate, sodium hydrogen phosphate, calcium carbonate, etc.

--Cellulose or cellulose derivatives--
The cellulose or cellulose derivative used as the excipient is contained to exhibit functions such as improving the tablet physical properties (moldability, disintegrability) immediately after production, promoting the dissolution of functional ingredients, improving the dissolution retention rate of functional ingredients, improving storage stability, suppressing tableting problems, etc. In particular, it is useful for exhibiting the functions of improving the tablet physical properties (moldability, disintegrability, dissolution), storage stability, and the dissolution retention rate of functional ingredients immediately after production, and the function of suppressing tableting problems.

The cellulose or cellulose derivative used as the excipient is not particularly limited and can be selected appropriately depending on the purpose. Examples include crystalline cellulose, methyl cellulose, microfibrous cellulose, hydroxypropyl cellulose, and ethyl cellulose. Among these, crystalline cellulose is preferred because of its excellent moldability and disintegration properties.

When the cellulose or cellulose derivative used as the excipient is crystalline cellulose, the bulk density of the crystalline cellulose is preferably 0.15 g/cm ³ or more and 0.40 g/cm ³ or less, and more preferably 0.20 g/cm ³ or more and 0.35 g/cm ³ or less.
The bulk density can be measured by the method described in "Microcrystalline Cellulose" in the Seventeenth Edition of the Japanese Pharmacopoeia using a Scott volume meter.

The average particle size of the cellulose or cellulose derivative used as the excipient is not particularly limited and can be selected appropriately depending on the purpose, but is preferably 20 μm or more and 180 μm or less, and more preferably 40 μm or more and 150 μm or less. The average particle size can be determined in the same manner as for (A) lactoferrin.

The content of the cellulose or cellulose derivative used as the excipient in the enteric coated solid preparation is preferably 15% by mass or more and 80% by mass or less, more preferably 20% by mass or more and 65% by mass or less, and even more preferably 25% by mass or more and 45% by mass or less.
When the content of the cellulose or cellulose derivative used as the excipient in the enteric coated solid preparation is 15% by mass or more, moldability, tablet disintegration property, dissolution property of the functional ingredient, disintegration property maintenance rate, and dissolution property maintenance rate are improved, and tableting problems are less likely to occur, which is preferable.
When the content of the cellulose or cellulose derivative used as the excipient in the enteric coated solid preparation is 80% by mass or less, sufficient hardness and good disintegrability can be obtained. In addition, the thickness can be reduced, which is preferable because the dosage is good and the mixed powder is less likely to adhere to the tablet press.

--Sugar alcohols--
The sugar alcohol is contained to exhibit functions such as improving tablet moldability, tablet physical properties (moldability, disintegrability), storage stability, and dissolution of functional ingredients immediately after production, as well as improving the dissolution maintenance rate of the functional ingredients, etc. In particular, the sugar alcohol is useful for exhibiting functions such as improving tablet physical properties (disintegrability), storage stability, and dissolution of functional ingredients immediately after production, as well as improving the dissolution maintenance rate of the functional ingredients.

The sugar alcohol is not particularly limited and can be selected appropriately depending on the purpose. Examples include maltitol, erythritol, xylitol, fructose, sucrose, glucose, sorbitol, and lactitol.

The average particle size of the sugar alcohol is not particularly limited and can be selected appropriately depending on the purpose, but is preferably 50 μm or more and 400 μm or less, and more preferably 200 μm or more and 300 μm or less. The average particle size can be determined in the same manner as for (A) lactoferrin.

The content of the sugar alcohol in the enteric coated solid preparation is preferably 5% by mass or more and 50% by mass or less, more preferably 10% by mass or more and 40% by mass or less, and even more preferably 12% by mass or more and 30% by mass or less.
When the content of the sugar alcohol in the enteric coated solid preparation is 5% by mass or more, moldability, disintegration property of the tablet, dissolution property of the functional ingredient, maintenance rate of disintegration property, and maintenance rate of dissolution property are improved, which is preferable.
When the content of the sugar alcohol in the enteric coated solid preparation is 50% by mass or less, sufficient hardness and good disintegrability can be obtained, and resistance to gastric juice can be ensured, which is preferable.

-Disintegrant-
The disintegrant is included to promote disintegration of the tablet.

The disintegrant is not particularly limited and can be selected appropriately depending on the purpose. Examples include carboxymethylcellulose (CMC) or its alkali metal salts, partially pregelatinized starch, etc. Among these, from the viewpoints of tablet disintegration property and dissolution property of functional ingredients, carboxymethylcellulose (CMC) or its alkali metal salts are preferred, and carboxymethylcellulose calcium is more preferred.

The content of the disintegrant is not particularly limited and can be appropriately selected depending on the purpose, but is preferably 0.1% by mass or more and 5% by mass or less, and more preferably 1% by mass or more and 2% by mass or less, in the enteric coated solid preparation.
When the content of the disintegrant in the enteric coated solid preparation is 0.1% by mass or more, the disintegration property of the tablet and the dissolution property of the functional ingredient are improved, which is preferable.
It is preferable that the content of the disintegrant in the enteric coated solid preparation is 5% by mass or less, since this reduces the delay in disintegration of the tablet during storage.

- Superplasticizer -
The fluidizing agent is contained to promote fluidity by preventing adhesion between powder particles, which is advantageous because the promotion of fluidity by the fluidizing agent reduces variation in tablet mass.

The fluidizing agent is not particularly limited and can be selected appropriately depending on the purpose. Examples include fine silicon dioxide (silica), light anhydrous silicic acid, calcium silicate, hydrous fine silicic acid powder, and talc.

When the fluidizing agent is silicon dioxide, the average particle size of the silicon dioxide is preferably 0.5 μm or more and 10 μm or less. The average particle size can be determined in the same manner as for (A) lactoferrin.
The silicon dioxide is preferably porous fine particle silicon dioxide.

The content of the fluidizer is not particularly limited and can be appropriately selected depending on the purpose, but from the viewpoint of suppressing variation in tablet mass, it is preferably 0.1% by mass or more and 2.0% by mass or less in the enteric coated solid preparation.
When the content of the fluidizing agent in the enteric coated solid preparation is 0.1% by mass or more, the fluidity of the mixed powder is improved and the variation in tablet mass is suppressed, which is preferable.
If the content of the fluidizing agent in the enteric coated solid preparation is 2.0% by mass or less, the hardness of the tablet does not become too high, which is preferable.

-lubricant-
The lubricant is added to prevent metal adhesion by reducing friction between the metal equipment and the powder particles, which is advantageous because it suppresses tableting problems.

The lubricant is not particularly limited and can be selected appropriately depending on the purpose. Examples include metal stearates and sucrose fatty acid esters. Of these, calcium stearate and magnesium stearate are preferred from the viewpoint of suppressing tableting problems (binding, sticking, and picking).

The content of the lubricant is not particularly limited and can be appropriately selected depending on the purpose. From the viewpoint of preventing adhesion to a tablet press and suppressing tableting problems (binding, sticking, picking), the content of the lubricant in the enteric coated solid preparation is preferably 0.01% by mass or more and 10% by mass or less, more preferably 0.05% by mass or more and 5.0% by mass or less, and even more preferably 0.10% by mass or more and 3.0% by mass or less.
When the content of the lubricant in the enteric coated solid preparation is 0.01% by mass or more, adhesion to the tablet press is prevented, and tableting problems (binding, sticking, picking) are less likely to occur, which is preferable.
If the content of the lubricant in the enteric coated solid preparation is 10% by mass or less, the tablet moldability, disintegrability, and dissolution properties are improved, which is preferable.

- Binder -
The binder is included to control the hardness of the tablet and the disintegration time of the tablet.

The binder is not particularly limited and can be selected appropriately depending on the purpose. From the viewpoints of preventing tableting problems (capping, lamination), tablet physical properties (hardness, disintegration time), and storage stability of tablet physical properties, examples of binders include starch such as corn starch, gum arabic, gelatin, sodium alginate, polyvinylpyrrolidone, polyvinyl alcohol, and hydroxypropyl methylcellulose (hereinafter sometimes referred to as "HPMC").

The content of the binder is not particularly limited and can be appropriately selected depending on the purpose. From the viewpoints of improving tablet formability, suppressing tableting problems, and improving the storage stability of the tablet physical properties, the content of the binder in the enteric coated solid preparation is preferably 0.01% by mass or more and 30% by mass or less, and more preferably 0.1% by mass or more and 10% by mass or less.
If the content of the binder in the enteric coated solid preparation is 0.01% by mass or more, the tablet moldability is improved and tableting problems are less likely to occur, which is preferable.
When the content of the binder in the enteric coated solid preparation is 30% by mass or less, the mixed powder is less likely to adhere to the tablet press, and the disintegration property of the tablet is improved, which is preferable.

- Coating agent -
The enteric coated solid preparation of the present invention may be a coated tablet provided with a coating layer containing a coating agent in order to improve ease of administration and mask the plain tablet (mainly the bitterness, odor, etc. caused by the functional ingredients).
The coating layer may contain a plasticizer in addition to the coating agent.
When the enteric coated solid preparation of the present invention is a coated tablet, the above-mentioned coating layer is present on the surface of the uncoated tablet.

The coating agent is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include cellulose, cellulose derivatives, methacrylic acid-based polymer compounds, shellac, zein, polyuronic acid and its salts, fucoidan and its salts, carrageenan and its salts, etc. Among these, HPMC, shellac, zein, polyuronic acid and its salts (particularly alginates), and pectin and its salts are preferred.
These may be used alone or in combination of two or more.

The cellulose derivative used in the coating agent is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include HPMC.
The polyuronic acid used in the coating agent is not particularly limited, and can be appropriately selected depending on the purpose, as long as it is a polymer of uronic acid such as guluronic acid, mannuronic acid, galacturonic acid, or glucuronic acid. Examples include alginic acid (a polymer of guluronic acid and mannuronic acid) and pectin (a polymer of galacturonic acid).

The plasticizer used in conjunction with the coating agent is not particularly limited and can be selected appropriately depending on the purpose. Examples include glycerin, polyethylene glycol, and sucrose fatty acid esters. Of these, glycerin and sucrose fatty acid esters are preferred.

When the enteric coated solid preparation of the present invention is a coated tablet, the content of the coating agent is not particularly limited and can be appropriately selected depending on the purpose. From the viewpoint of improving the ease of taking the tablet and masking the bitterness and odor derived from the functional ingredient, the content of the coating agent is preferably 0.5% by mass or more and 15% by mass or less, and more preferably 1% by mass or more and 10% by mass or less, per coated tablet.
When the content of the coating agent is 0.5% by mass or more per coated tablet, it is preferable because it improves ease of administration and masks the bitter taste and odor derived from the functional ingredients.
When the content of the coating agent is 15% by mass or less per coated tablet, the disintegration and dissolution properties of the tablet are good.

When the enteric coated solid preparation of the present invention is a coated tablet, the content of the coating agent is not particularly limited and can be appropriately selected depending on the purpose. However, from the viewpoint of improving the ease of taking the tablet and masking the bitterness and odor derived from the functional ingredient, the content of the coating agent in the enteric coated solid preparation is preferably 0.5% by mass or more and 15% by mass or less, and more preferably 1% by mass or more and 10% by mass or less.
When the content of the coating agent in the enteric coated solid preparation is 0.5% by mass or more, it is preferable because the ease of administration is improved and the bitter taste and odor derived from the functional ingredients are masked.
When the content of the coating agent in the enteric coated solid preparation is 15% by mass or less, the disintegration property and dissolution property of the tablet are good.

The content of the plasticizer used in the coating agent is not particularly limited and can be selected appropriately depending on the purpose. However, from the viewpoint of improving the disintegration and dissolution properties of the coated tablet, it is preferably 0.05% by mass or more per coated tablet. Furthermore, from the viewpoint of exerting the coating effect, the content of the plasticizer used in the coating agent is preferably 10% by mass or less per coated tablet, and more preferably 0.1% by mass or more and 5% by mass or less.

The content of the plasticizer used in the coating agent is not particularly limited and can be selected appropriately depending on the purpose. However, from the viewpoint of improving the disintegration and dissolution properties of the coated tablet, it is preferably 0.05% by mass or more in the enteric-coated solid formulation. Furthermore, from the viewpoint of exerting the coating effect, the content of the plasticizer used in the coating agent is preferably 10% by mass or less in the enteric-coated solid formulation, and more preferably 0.1% by mass or more and 5% by mass or less.

-Enteric coating agent-
The enteric coated solid preparation of the present invention may be an enteric coated tablet provided with a coating layer containing an enteric coating agent in order to inhibit the decomposition of ingredients that are susceptible to deterioration by gastric acid.
The enteric coating layer may contain a plasticizer in addition to the enteric coating agent.
When the enteric coated solid preparation of the present invention is an enteric coated tablet, the tablet has an enteric coating layer on the outside.

The enteric coating agent is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include methacrylic acid polymer compounds, shellac, zein, polyuronic acid and its salts, fucoidan and its salts, and carrageenan and its salts. Among these, shellac, zein, polyuronic acid and its salts (particularly alginates), and pectin and its salts are preferred. These may be used alone or in combination of two or more.
The polyuronic acid used in the enteric coating agent is not particularly limited as long as it is a polymer of uronic acid such as guluronic acid, mannuronic acid, galacturonic acid, or glucuronic acid, and examples thereof include alginic acid (a polymer of guluronic acid and mannuronic acid) and pectin (a polymer of galacturonic acid).
The enteric coating agent may be used in combination with a cellulose derivative (HPMC, etc.) exemplified as the coating agent.
The enteric coating agent may be used in combination with the coating agent.

When the enteric coated solid preparation of the present invention is an enteric coated tablet, the content of the enteric coating agent is not particularly limited and can be appropriately selected depending on the purpose. From the viewpoint of preventing the degradation of the functional ingredient in the stomach, the content is preferably 1% by mass or more and 30% by mass or less, and more preferably 1% by mass or more and 15% by mass or less, per enteric coated tablet.
It is preferable that the content of the enteric coating agent is 1% by mass or more per enteric coated tablet, since this can prevent the functional ingredient from being decomposed in the stomach.
When the content of the enteric coating agent is 30% by mass or less per one enteric coated tablet, the dissolution property in the intestinal environment is good.

When the enteric coated solid preparation of the present invention is an enteric coated tablet, the content of the enteric coating agent is not particularly limited and can be appropriately selected depending on the purpose. From the viewpoint of preventing the degradation of the functional ingredient in the stomach, the content of the enteric coating agent in the enteric coated solid preparation is preferably 0.1% by mass or more and 30% by mass or less, and more preferably 0.1% by mass or more and 15% by mass or less.
The content of the enteric coating agent in the enteric solid preparation is preferably 0.1% by mass or more, since this can prevent the functional ingredient from being decomposed in the stomach.
When the content of the enteric coating agent in the enteric solid preparation is 30% by mass or less, the dissolution property in the intestinal environment is good.
When the enteric coating agent is used in combination with a cellulose derivative (HPMC, etc.) exemplified as the coating agent, it is preferable that the amount of the coating agent be 0.1 parts by mass or more and 5 parts by mass or less per 1 part by mass of the enteric coating agent.

The plasticizer used in the enteric coating agent is not particularly limited and can be selected appropriately depending on the purpose. Examples include glycerin, polyethylene glycol, and sucrose fatty acid esters. Of these, glycerin and sucrose fatty acid esters are preferred.

The content of the plasticizer used in the enteric coating agent is not particularly limited and can be selected appropriately depending on the purpose. However, from the viewpoint of improving the disintegration and dissolution properties of the enteric coated tablet, it is preferably 0.05% by mass or more per one enteric coated tablet. Furthermore, from the viewpoint of exerting the coating effect, the content of the plasticizer used in the enteric coating agent is preferably 10% by mass or less per one enteric coated tablet, and more preferably 0.1% by mass or more and 5% by mass or less.

The content of the plasticizer used in the enteric coating agent is not particularly limited and can be selected appropriately depending on the purpose. However, from the viewpoint of improving the disintegration and dissolution properties of the enteric coated tablet, the content is preferably 0.05% by mass or more in the enteric coated solid formulation. Furthermore, from the viewpoint of exerting the coating effect, the content of the plasticizer used in the enteric coating agent is preferably 10% by mass or less in the enteric coated solid formulation, and more preferably 0.1% by mass or more and 5% by mass or less.

When the enteric coated solid preparation of the present invention is an enteric coated tablet, it is preferable to provide a pre-coating layer between the plain tablet and the enteric coating layer from the viewpoint of preventing a decrease in gastric juice resistance. Furthermore, an outermost layer may be provided outside the enteric coating layer to improve appearance, mouthfeel, and ease of administration.
Components contained in the pre-coating layer and the outermost layer include the coating agent, plasticizers used in the coating agent, etc. Among these, it is preferable to use the same components as the coating agent.
When the enteric coated solid preparation of the present invention has the pre-coating layer, the content of the pre-coating agent is not particularly limited and can be appropriately selected depending on the purpose, but it is preferably 0.1% by mass or more and 5% by mass or less in the enteric coated solid preparation.
When the enteric coated solid preparation of the present invention has the outermost layer, the content of the pre-coating agent is not particularly limited and can be appropriately selected depending on the purpose, but it is preferably 0.1% by mass or more and 5% by mass or less in the enteric coated solid preparation.

The dosage form of the enteric coated solid preparation of the present invention is not particularly limited and can be selected appropriately depending on the purpose. Examples include tablets (sugar-coated tablets, coated tablets, enteric-coated tablets, etc.), pills (sugar-coated tablets, coated tablets, enteric-coated tablets), and hard capsules (coated hard capsules, enteric-coated hard capsules). Of these, coated tablets and enteric-coated tablets are preferred because they allow for easier enjoyment of the effects of the present invention.

When the enteric coated solid preparation of the present invention is a tablet, the size of the tablet is not particularly limited and can be appropriately selected depending on the purpose. However, from the viewpoint of ease of handling and swallowing of the tablet, the diameter of the tablet is preferably 5 mmφ or more and 15 mmφ or less.
The thickness of the tablet is not particularly limited and can be appropriately selected depending on the purpose, but is preferably 3 mm or more and 10 mm or less.

When the enteric coated solid preparation of the present invention is a tablet, the mass of each tablet is not particularly limited and can be appropriately selected depending on the purpose, but is preferably 150 mg or more and 500 mg or less, and more preferably 250 mg or more and 400 mg or less.
A tablet mass of 150 mg or more is preferable because the required amount of functional ingredients can be blended in. A tablet mass of 500 mg or less is preferable because it improves ease of administration.

When the enteric coated solid formulation of the present invention is a tablet, the shape of the tablet is not particularly limited and can be selected appropriately depending on the purpose, but square flat tablets, round flat tablets, rounded R tablets, or two-stage R tablets are preferred.

When the enteric coated solid preparation of the present invention is a tablet, the structure of the tablet is not particularly limited and can be appropriately selected depending on the purpose, and may be a single-layer structure (single-layer tablet) or a laminated structure (laminated tablet). Among these, from the viewpoint of more easily enjoying the effects of the present invention, it is preferable to have a layer in which the component (A), the component (B), and the component (C) are used in combination (i.e., the component (A), the component (B), and the component (C) are present in the same layer). In this specification, the "layer in which the component (A), the component (B), and the component (C) are used in combination" may be referred to as the "active ingredient-containing layer."
When the enteric coated solid preparation of the present invention is a single-layer tablet, the enteric coated solid preparation is composed of an active ingredient-containing layer.When the enteric coated solid preparation of the present invention is a layered tablet, the enteric coated solid preparation is composed of an active ingredient-containing layer and a layer other than the active ingredient-containing layer (an optional layer).

When the enteric coated solid preparation of the present invention is a hard capsule, the size of the hard capsule is not particularly limited and can be appropriately selected depending on the purpose. The material of the capsule for the hard capsule is not particularly limited and can be appropriately selected depending on the purpose, and examples of materials that can be used include gelatin, hydroxypropyl methylcellulose, and pullulan.
Examples of the filler filled into the hard capsule include a mixed powder containing the component (A), the component (B), and the component (C), and granules obtained by granulation.

The dosage form of the enteric coated solid preparation of the present invention is not particularly limited and can be appropriately selected depending on the purpose. For example, it is preferably a conventional oral, ie, swallowed, tablet.
In the present invention, whether or not a preparation is an enteric coated solid preparation is determined by the disintegration test method of the Japanese Pharmacopoeia, 17th Edition.

Examples of the present invention will be described below, but the present invention is not limited to these examples. The contents of each component described in the examples and comparative examples are shown in "parts by mass" and are all values converted into pure contents.
The lactoferrin composition of the uncoated lactoferrin tablets in the examples of the present invention is the content of lactoferrin powder (raw material) (also listed is the pure lactoferrin content), and the content in the solid preparation is the content as pure lactoferrin. The pure lactoferrin content in lactoferrin powder is calculated according to the lactoferrin (in a dried product) quantification method and loss on drying (105°C, 5 hours) in the Japanese Food Additives Official Standards, 9th Edition, "Lactoferrin Concentrate" according to the following formula (5):
Pure lactoferrin content = lactoferrin content (in dry matter) (%) × (100 - loss on drying (%)) / 100 (5)

(Preparation of uncoated tablets)
Each of the raw tablet materials in Tables 2 to 7 was weighed and mixed so that the total mass of the mixed powder was 5 kg, and tableted using a rotary tablet press (LIBLA 2: manufactured by Kikusui Seisakusho) under the following conditions.
- Mortar and pestle: φ9.0 mm, two-stage R shape (R1 = 3.4 mm, R2 = 10 mm, land part (distance from the side of the tablet to the rising part of the R) = 1.4 mm)
・Feeder agitator blade rotation speed: 60 rpm
・Tablet pressure: 8kN or more and 11kN or less

(Preparation of first layer coating liquid)
The following materials were mixed to obtain a first layer coating liquid.
Hydroxypropyl methylcellulose (HPMC): 6% by mass
Glycerin: 2% by mass
Ion-exchanged water: 92% by mass

(Preparation of second layer coating liquid)
The following materials were mixed to obtain a second layer coating liquid.
Hydroxypropyl methylcellulose (HPMC): 2.1% by mass
Glycerin: 1.1% by mass
Sodium alginate: 3.2% by mass
Talc: 0.5% by mass
・Fine silicon dioxide: 0.5% by mass
Ion-exchanged water: 93.1% by mass

(Preparation of coated tablets)
Using a pan rotary coating machine (HiCoater FZ-Lab, manufactured by Freund Corporation), 33 g of the first layer coating solution (20°C) was sprayed onto 670 plain tablets at an average rate of 2 g/min, and coating was performed at a product temperature of approximately 50°C. Subsequently, 165 g of the second layer coating solution (60°C) was sprayed at an average rate of 2 g/min, and after spraying, the tablets were dried at approximately 45°C for 2 minutes to obtain enteric-coated solid preparations (coated tablets) of Examples 1 to 21 and Comparative Examples 1 to 4. The mass of the solid content derived from each coating layer per tablet is shown below.
<First Coating Layer (Solid Content by Mass)>
Hydroxypropyl methylcellulose (HPMC): 2.9% by mass
Glycerin: 1.0% by mass
<Second Coating Layer (Solid Content by Mass)>
Hydroxypropyl methylcellulose (HPMC): 4.6% by mass
Glycerin: 2.3% by mass
Sodium alginate: 7.1% by mass
・Fine silicon dioxide: 1.1% by mass
Talc: 1.1% by mass

The enteric-coated solid formulations of Examples 1 to 21 and Comparative Examples 1 to 4 were evaluated for "tabletting problems," "initial dissolution rate," "dissolution retention rate," and "acid resistance over time" using the following methods. The results are shown in Tables 2 to 7 below.

[Tableting problems]
When tableting was performed under the above-mentioned conditions for the enteric coated solid preparations of Examples 1 to 21 and Comparative Examples 1 to 4, 20 tablets were sampled from each sample. Of the obtained tablets, those with vertical lines (binding) on the side were visually inspected and counted. Evaluation was performed according to the following criteria.
[Criteria for determining tableting problems]
3 points: No binding observed 2 points: Binding observed with 1-2 tablets 1 point: Binding observed with 3-5 tablets 0 point: Binding observed with 6 or more tablets

<Dissolution test>
The lactoferrin dissolution rate (%) was measured according to the tablet dissolution test method described in the 17th edition of the Japanese Pharmacopoeia. Specifically, lactoferrin was dissolved into the test solution using the paddle method at 50 revolutions per minute. The test solution used was the second dissolution test solution with a pH of approximately 6.8, at 900 mL per tablet. Two hours after the start of the test, the test solution was sampled, and the amount of lactoferrin dissolved in the test solution was quantified to calculate the dissolution rate.
Furthermore, a test was also conducted using the first dissolution test fluid at a pH of about 1.2 to confirm that the coating had been formed normally. Therefore, if it is confirmed that the tablets of this example have dissolution properties in the second dissolution test fluid, it will be confirmed that the tablets of this example are bioavailable as enteric-coated solid preparations.
The amount of lactoferrin eluted into the test solution was determined as follows: First, 75 mg of lactoferrin standard (lactoferrin derived from milk, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was placed in a 50 mL volumetric flask, and the flask was then filled up to the specified volume with elution test fluid 2 to obtain a 1/1 standard solution. This standard solution was further diluted with elution test fluid 2 to prepare 1/5, 1/20, and 1/50 standard solutions. Lactoferrin in each standard solution was detected by HPLC. A calibration curve was created by determining the peak area of lactoferrin in each standard solution.
Next, lactoferrin in the test solutions collected as described above was detected by HPLC to determine the lactoferrin peak area. The lactoferrin concentration in each test solution was determined from the peak area based on the calibration curve, and the amount of lactoferrin eluted in the test solution was quantified.
The purity of the lactoferrin standard was quoted from the content (HPLC) value listed in the test report issued by Fujifilm Wako Pure Chemical Industries, Ltd.
The HPLC conditions were as follows:
- Detector: ultraviolet absorption photometer (measurement wavelength: 280 nm)
Column packing material: 5 μm butylated polyvinyl alcohol polymer gel for liquid chromatography (Shodex Asahipak C4P-50 4D)
- Column tube: stainless steel tube with an inner diameter of 4.6 mm and a length of 15 cm - Guard column: Shodex Asahipak C4P-50G 4A
- Column temperature: 35°C
Mobile phase A: 0.03 w/v weight % trifluoroacetic acid-containing acetonitrile/sodium chloride solution (3 → 100) mixture (10:90)
Mobile phase B: 0.03 w/v weight % trifluoroacetic acid-containing acetonitrile/sodium chloride solution (3 → 100) mixture (50:50)
- Concentration gradient: A linear concentration gradient from A:B (50:50) to A:B (0:100) is applied over 25 minutes.
-Flow rate: 0.8mL/min

[Initial lactoferrin dissolution rate]
The enteric-coated solid preparations of Examples 1 to 21 and Comparative Examples 1 to 4 were subjected to the dissolution test described above to determine the initial lactoferrin dissolution amount from the enteric-coated solid preparations. To obtain the initial lactoferrin dissolution rate, tablets were used within 7 days after tableting. The test was performed three times each, and the initial lactoferrin dissolution rate (%) was calculated by dividing the measured initial lactoferrin dissolution amount by the theoretical amount of lactoferrin (converted to pure content) calculated from the weighed amount of lactoferrin. The average initial lactoferrin dissolution rate was then calculated. Evaluation was performed according to the following criteria. None of the formulations in these Examples had an initial dissolution rate of less than 75%.
[Criteria for determining initial lactoferrin dissolution rate]
3 points: Initial lactoferrin dissolution rate is 96% or more. 2 points: Initial lactoferrin dissolution rate is 93% or more but less than 96%. 1 point: Initial lactoferrin dissolution rate is 90% or more but less than 93%. 0 point: Initial lactoferrin dissolution rate is less than 90%.

[Lactoferrin elution retention rate]
The enteric-coated solid preparations of Examples 1 to 21 and Comparative Examples 1 to 4 were filled into plastic bottles and stored at 60°C for 12 days, after which the above-mentioned dissolution test was conducted to determine the amount of lactoferrin eluted from the enteric-coated solid preparations. The test was conducted three times each, and the average lactoferrin elution retention rate was calculated from the measured values of the lactoferrin elution rate. Evaluation was conducted according to the following criteria. The lactoferrin elution retention rate (%) was calculated according to the following formula (6).
Lactoferrin elution retention rate (%) = 100 × {(elution amount after storage (mg)) / (initial elution amount (mg))} (6) [criteria for determining lactoferrin elution retention rate]
3 points: 85% or more 2 points: 78% or more but less than 85% 1 point: 71% or more but less than 78% 0 points: Less than 71%

[Gastric juice resistance over time]
The enteric-coated solid preparations of Examples 1 to 21 and Comparative Examples 1 to 4 were filled into plastic bottles and stored at 60°C for 12 days. A dissolution test was then conducted using the first dissolution test fluid (pH 1.2) in the dissolution test for tablets described in the Japanese Pharmacopoeia, Seventeenth Edition, in accordance with the general test method described in the Japanese Pharmacopoeia, Seventeenth Edition, by the paddle method (rotation speed: 50 rpm). The amount of lactoferrin in the dissolution test fluid sampled after 2 hours and 30 minutes was quantified according to the same test method. Six tablets were tested for each test. Evaluation was based on the following criteria:
[Criteria for determining gastric juice resistance over time]
3 points: Lactoferrin was below the detection limit for all 6 tablets. 2 points: 1-2 tablets had a lactoferrin elution rate of 5% or more. 1 point: 3-4 tablets had a lactoferrin elution rate of 5% or more. 0 point: 5 or more tablets had a lactoferrin elution rate of 5% or more.

[Overall evaluation as an enteric-coated solid preparation]
A product with a total score of 10 or more in the evaluation of the tableting trouble, the evaluation of the initial lactoferrin dissolution rate, the evaluation of the lactoferrin dissolution maintenance rate, and the evaluation of gastric juice resistance after time was deemed to be acceptable.

Details of the components used in Examples 1 to 21 and Comparative Examples 1 to 4 are shown in Table 8 below.

(*1): Lacticaseibacillus casei (Lactobacillus casei, manufactured by THT, Belgium, sold by Seti Co., Ltd.) was anaerobically cultured in MRS medium (Oxoid), harvested, sterilized by high-pressure steam at 121°C for 20 minutes, and then freeze-dried. 2 mg of this dried product was prepared to correspond to 40 billion lactic acid bacteria.
(*2): Lacticaseibacillus paracasei (Lactobacillus paracasei, manufactured by THT, Belgium, sold by Seti Co., Ltd.) was anaerobically cultured in MRS medium (Oxoid), collected, sterilized by high-pressure steam at 121°C for 20 minutes, and then freeze-dried. 2 mg of this dried product was prepared to correspond to 40 billion lactic acid bacteria.
(*3): Bifidobacterium longum (Bifidobacterium longum, Morinaga Milk Industry's Bifidus Drinking Yogurt) was used as a sample. The sample was anaerobically cultured in MRS medium (Oxoid), then collected and freeze-dried. 20 mg of this dried product was prepared to contain 40 billion bifidobacteria.
(*4): Lacticaseibacillus casei (Lactobacillus casei, manufactured by THT, Belgium, sold by Seti Co., Ltd.) was anaerobically cultured in MRS medium (manufactured by Oxoid), harvested, and sterilized by high-pressure steam at 121°C for 20 minutes. An equal amount of DE11.6 dextrin (Sandec #100, manufactured by Sanwa Starch Co., Ltd.) was added as a dilution excipient to the lactic acid bacteria, and the mixture was freeze-dried. The dried product was prepared so that 40 billion lactic acid bacteria were present in 40 mg of the dried product (20 mg of the 40 mg was dextrin).
(*5): Bifidobacterium longum (Bifidobacterium longum, Morinaga Milk Industry's Bifidus Drinking Yogurt) was used as a sample. After anaerobically culturing in MRS medium (Oxoid), the bacteria were harvested and an equal amount of dextrin with a DE of 11.6 (Sanwa Starch Industry's Sandec #100) was added as a dilution excipient to the lactic acid bacteria, followed by freeze-drying. 40 mg of this dried product was prepared to contain 40 billion bifidobacteria (20 mg of the 40 mg was dextrin).
(*6): The DE value of dextrin is the DE value recorded on the performance report of the purchased raw material (the DE value was measured by the measurement method described in the Central Customs Analysis Laboratory, "Quantitative Analysis Method for Reducing Sugars in Dextrin" in the report of the Central Customs Analysis Laboratory mentioned above (II)).
(*7): The dextrin having a DE value of 35 in Comparative Example 4 was a mixture of 10 mg of dextrin (GLUCIDEXIT 19) having a DE value of 28.9 and 10 mg of dextrin (GLUCIDEXIT 47) having a DE value of 44.

Claims (2)

(A) lactoferrin, (B) lactic acid bacteria, and (C) dextrin having a dextrose equivalent (DE) of 10 or more and 30 or less;
an enteric coating layer on the surface of the uncoated tablet;
An enteric solid preparation having a pre-coating layer between the uncoated tablet and the enteric coating layer,
the pre-coating layer and the enteric coating layer comprise an enteric coating agent and a plasticizer;
the enteric coating agent contains at least one selected from cellulose, hydroxypropyl methylcellulose, a methacrylic acid-based polymer compound, shellac, zein, polyuronic acid and its salt, fucoidan and its salt, and carrageenan and its salt;
the plasticizer includes at least one selected from glycerin, polyethylene glycol, and sucrose fatty acid ester;
An enteric coated solid preparation , wherein the mass ratio of the content of the component (C) to the content of the component (A) [(C)/(A)] is 0.1 or more and 0.36 or less . the pre-coating layer comprises hydroxypropyl methylcellulose and glycerin;
2. The enteric coated solid dosage form according to claim 1, wherein the enteric coating layer comprises hydroxypropyl methylcellulose, glycerin, sodium alginate, fine silicon dioxide, and talc .