Artificial saliva, artificial saliva bursting beads, preparation method thereof, and application thereof in xerostomia

Abstract

The present invention belongs to the technical field of artificial saliva, and specifically discloses an artificial saliva, which is an aqueous solution containing cetylpyridinium chloride, HPMC, glycerol, electrolytes, sweeteners, preservatives, and pH regulators; wherein cetylpyridinium chloride is 0.01-0.12wt.%; HPMC is 0.2-2.5wt.%; and the content of glycerol is 1-5wt.%. In addition, the present invention also includes popping beads containing the artificial saliva and the preparation and application thereof. The artificial saliva of the present invention, based on the combination of the components, can synergistically improve the antibacterial and oral adhesion effects, and is innovatively prepared into a popping bead preparation, which can further improve its oral local application effect.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and in particular relates to the field of artificial saliva for treating xerostomia. Background Art

[0002] Saliva is a kind of body fluid secreted by oral mucosa that has the functions of moisturizing, lubricating, inhibiting bacteria, protecting the soft and hard tissues in the oral cavity and promoting digestion. It is one of the most important components in the oral cavity. The total amount of saliva secreted by a normal person every day is about 1000-1500 ml. Its composition includes more than 99% water; organic matter is mainly salivary amylase, mucin, lysozyme and mucopolysaccharide; inorganic ions are mainly potassium, sodium and calcium.

[0003] Xerostomia, also known as dry mouth, refers to the subjective feeling of dry mouth and the objective reduction of saliva secretion, including reduced saliva flow and changes in saliva composition. The prevalence of xerostomia is about 5.5% to 46%, and the prevalence increases with age. There are many causes of xerostomia, which generally include drugs, aging, radiotherapy, chemotherapy, and systemic diseases such as rheumatoid, endocrine, neurological, genetic, metabolic and infectious diseases. Qualitative and quantitative salivary secretion dysfunction leads to the feeling of dry mouth, which brings great discomfort and even pain to patients, has a great negative impact on the quality of life of patients, leads to changes in oral flora and saliva composition, and causes caries, halitosis, periodontitis, mucositis, and candidal infection.

[0004] If dry mouth is not properly treated, it will lead to more serious other symptoms and seriously affect the quality of life of the population. For those patients who endure dry mouth, it is difficult for them to get treatment and maintain the efficacy. According to the causes and symptoms of dry mouth, in order to prevent and correct the possible consequences of lack of natural saliva, treatment can be roughly divided into two categories: endogenous and exogenous. Endogenous methods include drug therapy, mechanical stimulation, gene therapy, etc., which stimulate the secretion of water, electrolytes and macromolecules by replacing or enhancing the function of salivary glands, or preventive protection against harmful factors such as ionizing radiation. Exogenous methods include local application of saliva substitutes to replace the lost or enhance the existing functions of natural saliva, such as drinking water, moisturizers, saliva substitutes, etc. When the symptoms of dry mouth are not obvious, patients will drink a lot of water. But water itself cannot provide sufficient hydration and lubrication, nor can it provide antibacterial properties. Artificial saliva contains substances of natural origin and has good biocompatibility. It can alleviate symptoms, replace frequent drinking of water, moisturize mucosa, and relieve oral discomfort. Its viscosity is higher than that of water, similar to that of natural saliva, which can provide tissue protection, facilitate speaking and eating, and alleviate the symptoms of dry mouth and reduce caries and mucosal inflammation.

[0005] In the prior art, there are some reports on artificial saliva containing dissolved inorganic salts, thickeners, flavoring agents and other ingredients. However, the oral adhesion ability, antibacterial properties, bioavailability and other aspects of the existing artificial saliva need to be improved. In addition, the existing artificial saliva is mainly solution-type and not very targeted, which can easily lead to compliance and is not easy to use for a long time. Summary of the invention

[0006] In view of the defects of the prior art, the first purpose of the present invention is to provide an artificial saliva, which aims to synergistically improve the antibacterial, adhesion and xerostomia treatment effects based on the combination of the new artificial saliva formula.

[0007] The second purpose of the present invention is to provide the use of the artificial saliva in xerostomia.

[0008] The third object of the present invention is to provide bursting beads (also referred to as bursting beads or gel beads in the present invention) encapsulating the artificial saliva, aiming to fill the technical gap of artificial saliva bursting beads, and through the combination of the artificial saliva and the bursting bead structure, synergistically improve the physicochemical properties and efficacy of the new artificial saliva preparation.

[0009] The fourth object of the present invention is to provide a method for preparing the artificial saliva bursting beads, aiming to provide a preparation method that can achieve artificial saliva encapsulation, improve encapsulation rate, improve bursting bead properties, oral adaptability and efficacy.

[0010] The fifth purpose of the present invention is to provide the use of the artificial saliva bursting beads in treating dry mouth.

[0011] An artificial saliva is an aqueous solution containing cetylpyridinium chloride, HPMC, glycerol, electrolytes, sweeteners, preservatives and pH regulators; wherein the cetylpyridinium chloride content is 0.01-0.12wt.%, the HPMC content is 0.2-2.5wt.%, and the glycerol content is 1-5wt.%.

[0012] The present invention provides a new artificial saliva formula. In the artificial saliva of the present invention, the combination of components such as cetylpyridinium chloride, HPMC, glycerol and the like and the content thereof can unexpectedly achieve synergy, significantly improve antibacterial properties, improve local adhesion ability, and improve the therapeutic effect of xerostomia.

[0013] In the present invention, the electrolyte is at least one of potassium chloride and sodium chloride; preferably, it contains potassium chloride and sodium chloride; more preferably, the weight ratio of potassium chloride to sodium chloride is 1 to 2:1.

[0014] Preferably, the sweetener is at least one of xylitol and steviol glycoside, preferably including xylitol and steviol glycoside.

[0015] Preferably, the preservative is at least one of sorbic acid and sodium benzoate, preferably sorbic acid and sodium benzoate.

[0016] Preferably, the pH regulator is at least one of potassium dihydrogen phosphate, sodium hydroxide, and potassium hydroxide;

[0017] Preferably, the molecular weight of HPMC is 100,000 to 1.5 million.

[0018] In the present invention, the weight percentage of each component of the artificial saliva is: cetylpyridinium chloride 0.08-0.12wt.%; HPMC 1.2-2.5wt.%; glycerol content is 1.5-5.5wt.%; electrolyte 0.16-0.25%; sweetener 2.4-3.6%; preservative: 0.1-0.3%, pH 6-8; the balance is water. In the present invention, the artificial saliva under the preferred ratio can not only synergistically improve the antibacterial performance and adhesion, but also facilitate the successful encapsulation, improve the physicochemical properties of the encapsulated popping beads, and improve their oral local release effect.

[0019] Preferably, the weight percentage of each component of the artificial saliva is: cetylpyridinium chloride 0.08-0.12wt.%; HPMC 1.6-1.8wt.%; glycerol content is 1-3wt.%; electrolyte 0.2-0.25%; sweetener 2.4-3.2%; preservative: 0.1-0.2%, pH 6-7; the balance is water. In the present invention, under the preferred ratio, it helps to ensure good synergistic antibacterial and oral local applicability, and also helps to unexpectedly further improve the encapsulation performance and avoid breakage. In addition, it also helps to improve the lithiation properties of the encapsulated material, making it suitable for oral local release.

[0020] The present invention also provides a method for preparing the artificial saliva, which is formed by compounding various components.

[0021] The preferred preparation steps are, for example, dispersing cetylpyridinium chloride, glycerol, electrolytes, sweeteners, preservatives, and pH adjusters in water, then adding HPMC, and then making up to volume with water to obtain the artificial saliva.

[0022] In the present invention, the compounding process is carried out under stirring, ultrasonic and other conditions.

[0023] The present invention also provides an application of the artificial saliva, which is used for preparing a medicine for preventing and / or treating xerostomia.

[0024] Preferably, the composition is used for preparing a medicament for local oral release for preventing and / or treating xerostomia.

[0025] The present invention also provides an artificial saliva bursting bead, comprising a wall material and a core material coated in the wall material, wherein the core material is the artificial saliva.

[0026] The present invention provides a new type of artificial saliva preparation of bursting beads, which fills the gap in the technology of artificial saliva bursting beads. In the present invention, thanks to the combination of the artificial saliva components and the bursting beads preparation type, unexpected synergy can be achieved, which can not only improve the atomization properties of the artificial saliva bursting beads, such as improving the stability and surface gloss of the artificial saliva bursting beads; in addition, it also helps to synergistically improve the antibacterial and drug efficacy properties of the artificial saliva bursting beads, not only that, but also improve the stability of artificial saliva and reduce toxic side effects.

[0027] Preferably, in the artificial saliva of the artificial saliva bursting beads core material, the weight percentage of each component in the artificial saliva as the core material is: cetylpyridinium chloride 0.08-0.12wt.%; HPMC 1.2-2.5wt.%; glycerol content is 1-5wt.%; electrolyte 0.16-0.25%; sweetener 2.4-3.6%; preservative: 0.1-0.3%, pH 6-8, and the balance is water. Further preferably, cetylpyridinium chloride 0.08-0.12wt.%; HPMC 1.6-1.8wt.%; glycerol content is 1-3wt.%; electrolyte 0.2-0.25%; sweetener 2.4-3.2%; preservative: 0.1-0.2%, pH 6-7; the balance is water;

[0028] In the present invention, the wall material is a cross-linked product of alginic acid and its salts;

[0029] Preferably, the wall material is a cross-linked product of sodium alginate in calcium salt.

[0030] The artificial saliva bursting beads of the present invention have a particle size of 12-16 mm and an artificial saliva content of 90%-98% by weight.

[0031] Preferably, the particle size of the artificial saliva bursting beads is 13-15 mm; the content of artificial saliva is 93-96 wt%.

[0032] The present invention also provides a method for preparing the artificial saliva bursting beads, which is prepared by reverse spherification, secondary gelation and combined modification process, and the steps include:

[0033] Step (1): adding a raw material solution containing artificial saliva and a calcium source dropwise into a wall material solution, performing a reverse spheroidization treatment, and then separating to obtain primary explosive beads;

[0034] Step (2): placing the primary bursting beads in a calcium source solution for secondary gelation, and then soaking and modifying them in glycerol to obtain the artificial saliva bursting beads;

[0035] The time of immersion modification treatment is 10 to 50 seconds.

[0036] Different contents of the popping beads have different encapsulation difficulties and need to overcome different encapsulation problems caused by different contents. For example, in order to fill the gap in the industry, the present invention successfully encapsulates artificial saliva for the first time in the industry, which needs to overcome many technical problems caused by artificial saliva components, such as difficulty in encapsulation, loose and fragile encapsulation structure, insufficient hardness and roundness, easy dehydration and wrinkling of the surface, unsatisfactory smoothness, bitterness caused by encapsulation means, and low encapsulation rate. In response to the problems existing in the preparation of bursting beads by encapsulating artificial saliva, the present invention has innovatively discovered through research that under the coordinated control of the components of artificial saliva, further combined with reverse spheroidization, two-stage gelation and glycerol immersion modification treatment, the encapsulation of artificial saliva can be successfully achieved. Not only that, it helps to improve the compactness of the encapsulated structure and reduce the breakage rate. In addition, it can also improve the spheroidization degree, reduce dehydration wrinkles, and improve smoothness. Not only that, it can also improve the encapsulation rate and avoid the problem of unsuitable taste caused by encapsulation. In addition, it can also synergistically improve the efficacy of the prepared artificial saliva bursting beads.

[0037] In the present invention, the combination of the artificial saliva components and the reverse spherification, secondary gelation and glycerol modification process is the key to successfully achieve artificial saliva encapsulation, solve process problems, and improve the properties and efficacy of the explosive bead preparation. On this basis, further controlling the content of the components in the artificial saliva helps to further synergistically improve the encapsulation physicochemical properties and improve the oral local release effect of the explosive beads.

[0038] In the present invention, the combination of the components of the artificial saliva can be effectively embedded into the structure of the wall material, improve the structural density, toughness and hardness of the popping beads, improve the storage stability, reduce the water loss rate, and improve the surface smoothness.

[0039] In the present invention, in order to obtain a better artificial saliva encapsulation effect, preferably, the artificial saliva is controlled to have 0.08-0.12wt.% of cetylpyridinium chloride; 1.2-2.5wt.% of HPMC; 1-5wt.% of glycerol; 0.16-0.25% of electrolyte; 2.4-3.6% of sweetener; 0.1-0.3% of preservative, pH 6-8, and the balance is water. It is further preferred to control the artificial saliva to have 0.08-0.12wt.% of cetylpyridinium chloride; 1.6-1.8wt.% of HPMC; 1-3wt.% of glycerol; 0.2-0.25% of electrolyte; 2.4-3.2% of sweetener; 0.1-0.2% of preservative, pH 6-7; and the balance is water. Combined with reverse spheroidization, secondary coagulation and glycerol modification processes, artificial popping beads with better physicochemical properties and better encapsulation rate are prepared.

[0040] The reverse spheroidization described in the present invention refers to a technology of dropping artificial saliva into a wall material solution for spheroidization. The present invention has found that the reverse spheroidization process can unexpectedly achieve the encapsulation of artificial saliva, and is beneficial to the coordination with other processes to improve the physicochemical properties and efficacy of artificial saliva bursting beads, such as improving their spheroidization degree, surface smoothness, structural density, hardness and toughness, and facilitating their local release in the oral cavity.

[0041] In the present invention, the calcium source is a water-soluble calcium salt, preferably at least one of calcium chloride, calcium lactate, calcium nitrate and calcium acetate.

[0042] Preferably, in step (1), the concentration of the calcium source in the raw material solution is 4% to 6%.

[0043] The alginate is sodium salt, potassium salt, etc. of alginic acid. Preferably, the concentration of the solute in the alginate solution is 0.25% to 1%.

[0044] Preferably, in step (2), the calcium source solution is an aqueous solution of a water-soluble calcium salt, preferably an aqueous solution of at least one of calcium chloride, calcium lactate, calcium nitrate and calcium acetate, and the concentration of the solute in the calcium source solution is 4% to 6%.

[0045] Preferably, the temperature of the secondary gelation process is 35-37° C. and the time is 4-10 min.

[0046] During the titration process, control the titration distance to 6-8 cm. After titration, harden for 4-10 minutes. Use ultrasonic and magnetic stirrers to stir / oscillate continuously to avoid sticking.

[0047] Preferably, the glycerol soaking modification treatment time is 15s to 30s, preferably 12 to 18s. In the present invention, under the preferred process parameters, the sphericity, encapsulation efficiency, surface smoothness, and physical and chemical properties suitable for oral local release of the prepared popping beads can be unexpectedly improved.

[0048] The present invention also provides an application of the artificial saliva bursting beads, which is used for preparing a medicine for preventing and / or treating dry mouth.

[0049] Beneficial effects:

[0050] 1. The present invention provides an artificial saliva formula, which is based on the combination of cetylpyridinium chloride, HPMC, glycerol and other ingredients in the formula, which can achieve synergy, synergistically improve antibacterial properties, and improve the adhesion of antibacterial drugs to the oral epithelial mucosa. In addition, further controlling the content of the ingredients is helpful to further improve the encapsulation rate and loading rate, and improve the physicochemical properties of the encapsulation.

[0051] 2. The present invention also provides an artificial saliva bursting bead dosage form. The present invention has found that, thanks to the combination of the artificial saliva components and the bursting bead dosage form, further synergy can be achieved to provide a new artificial saliva dosage form, improve the activity and stability of antibacterial drugs, synergistically improve the therapeutic effect of xerostomia, and reduce dependence.

[0052] 3. The present invention innovatively provides an artificial saliva encapsulation method based on artificial saliva formula, reverse spheroidization, secondary coagulation and glycerol-HPMC synergistic modification. Based on the combination of the above ingredients and processes, the present invention can achieve synergy and solve the problems of unsatisfactory preparation morphology and performance faced by the artificial saliva encapsulation process. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 : Product diagram of orthogonal experiment 4 of embodiment 2;

[0054] Figure 2 : The product image of orthogonal experiment 1 of Example 2 has certain tailing deformation, and the product shape is not as good as that of Example 2-Group 4;

[0055] Figure 3 Diameter and weight analysis diagram of the orthogonal experiment of Example 2;

[0056] Figure 4 It is the sphericity analysis diagram of the orthogonal experiment of Example 2;

[0057] Figure 5 It is the load capacity analysis diagram of the orthogonal experiment of Example 2;

[0058] Figure 6 This is the hardness-film analysis diagram of the orthogonal experiment of Example 2;

[0059] Figure 7 This is the product picture of Comparative Example A;

[0060] Figure 8 This is the product picture of group b of comparative example C;

[0061] Fig. 9 Comparison of the number of micronuclei in each dose group and the negative group with that in the positive group (red indicates micronuclei, magnification 1000×);

[0062] Fig.10 This is a specific experimental diagram of Example 4; DETAILED DESCRIPTION

[0063] Main materials and reagents

[0064]

[0065] Quality evaluation of artificial saliva gel beads containing cetylpyridinium chloride requires determination of the incoming content of cetylpyridinium chloride and microbiological and toxicological studies of the preparation.

[0066] The analytical methods for determining the content of active ingredients in compound preparations are mainly instrumental analysis, such as chromatography, optical analysis, chemical analysis (those that have been proven to be reliable can also be used). When measuring cetylpyridinium chloride, the selection of chromatographic conditions is diverse. This study selected a detection wavelength of 258nm and a chromatographic column of CN. 18 When the column was used, the chromatographic peak was severely tailed and the peak elution time was late. Acetonitrile was selected as the extraction solvent mainly because of its good peak shape, fewer impurity peaks, and little effect on the main peak (Xu Fanghui, Zhang Xianchong, Xiao Benyou, et al. Preparation and content determination of compound cetylpyridinium chloride gel [J]. Central South Pharmacy, 2017 (11): 1532-1536.).

[0067] At the same time, this analytical method needs to undergo a series of reliability demonstration tests. The specificity test is to prove that, except for the active ingredient, all other ingredients will not interfere with the determination of the active ingredient. The linearity of the regression equation is determined by using this method to determine a series of five simulated samples of known concentration (the multiple difference is mostly 5-10 times), and using the measurement signal (such as chromatographic peak area or peak height, etc.) to perform linear regression on its concentration or mass (correlation coefficient r≥0.999, n≥5). The recovery test is a type of "control test" and is expressed by the spiked recovery percentage (average value ±10.0%, n≥6), which is called "percent recovery rate" or "recovery rate". When the components of the sample analyzed in the recovery test are complex and not yet fully understood, a known amount of the component to be tested is added to the sample for determination to check whether the added component can be quantitatively recovered, and finally determine whether there is a systematic error in the analysis process. The repeatability test is to measure the simulated sample of known content six times in a row (RSD≤5.0%, n≥6). In the preparation stability determination, the appearance inspection should observe and record whether the preparation color changes, whether there are sediments or suspended matter (liquids), and whether the appearance properties are intact (tablets). Among them, the accelerated test method or accelerated method is to place the drug under simulated extreme climate conditions for stability research to determine the content of the active ingredient in the preparation. Its purpose is to explore the stability of the drug by accelerating the physical or chemical changes of the drug, and to provide the necessary basis for preparation design, packaging, transportation and storage. When the sample is stored at 37°C for 3 months and the decrease rate of the effective bactericidal component content is less than 10%, the shelf life can be set to 2 years; when stored at 54°C for 14 days, the decrease rate of the effective bactericidal component content is less than 10%, the shelf life can be set to 2 years (Ministry of Health of the People's Republic of China, Technical Specifications for Disinfection [S].).

[0068] According to regulations, all disinfectants should be tested for microbial killing (Ministry of Health of the People's Republic of China, Technical Specifications for Disinfection [S]). In simulated field tests or field tests, skin can be used to replace mucosa for mucosal preparations (Gao Dongqi. The Ministry of Health issued a new "Technical Specifications for Disinfection" [J]. Medical Animal Control, 2003, 19 (6): 384-384). The practical dose of the preparation for hand and skin disinfection is verified by measuring the killing effect of the preparation on natural skin surface bacteria. In order to ensure the safety of the preparation, in addition to strictly implementing the relevant technical regulations, relevant mandatory standards and / or the prohibition or restriction requirements of the relevant national departments on the content of each component or impurity (i.e., contaminant) in the preparation, corresponding safety and toxicology evaluations should also be carried out. The main purpose of safety research is to determine the possible pharmacological effects of the preparation that are related to human safety but are not expected; to evaluate the adverse reactions and / or pathophysiological effects observed in toxicological and / or clinical studies; to study the observed adverse reactions and / or speculate on the possible adverse reaction mechanisms of the preparation. Liu Mei, Tian Xin. Comparative study and implementation suggestions of "Technical Guidelines for Drug Safety Pharmacology Research" and "Technical Guidelines for Nonclinical Studies of Potential Effects of Drugs Prolonging QT Interval" with ICH S7A / S7B[J]. Modern Drugs and Clinics, 33(10):2745-2748.

[0069] Guang Hongmei, Wang Qingli, Wang Haixue. Introduction to the revision of the Technical Guidelines for Drug Irritation, Allergy and Hemolysis Studies[J]. Chinese Journal of Clinical Pharmacology, 2015, 31(6): 482-484.)

[0070] Part I: Artificial saliva composition and performance screening:

[0071] 1.1: Artificial saliva formula

[0072] Embodiment 1:

[0073] Artificial saliva is an aqueous solution containing an antibacterial agent, a thickener, glycerin, an electrolyte, a sweetener, a preservative, and a pH regulator. The preparation steps are: dispersing the antibacterial agent, glycerin, an electrolyte (sodium chloride and potassium chloride in a weight ratio of 2.11:3), a sweetener (sorbic acid and sodium benzoate in a weight ratio of 0.08:0.1), a preservative (sorbic acid and sodium benzoate in a weight ratio of 3:4), and a pH regulator (potassium dihydrogen phosphate) in water, then adding a thickener, and then diluting with water to obtain the artificial saliva.

[0074] Based on Example 1, the ingredients and proportions were further adjusted. The ingredients and proportions of each case are shown in Table 1:

[0075] Table 1

[0076]

[0077]

[0078] (1) Antibacterial performance data

[0079] The antibacterial activity of the artificial saliva provided in different cases (experiments 1 to 3) of Example 1 and comparative examples 1 to 5 was studied.

[0080] Common tests for determining the inhibitory effect of antimicrobial preparations on bacteria and fungi include the antibacterial ring test, the residual antibacterial effect test, and the minimum inhibitory concentration test. The principle of the antibacterial ring test is that after the antibacterial preparation is continuously dissolved, agar diffusion can form different concentration gradients. The preparation can determine whether the preparation has antibacterial ability based on the size of the ring.

[0081] Before the test, appropriate representatives of the neutralization test microorganisms should be selected for microbiological research. Among the bacterial propagules, Staphylococcus aureus ATCC 6538 is a representative of pyogenic cocci; Escherichia coli 8099 is a representative of intestinal bacteria; Pseudomonas aeruginosa ATCC15442 can be used as a representative of the most common isolated bacterial propagules in hospital infections; other bacteria, such as airborne bacteria, are represented by Staphylococcus albus 8032; pathogenic fungi are represented by Candida albicans ATCC10231 and Aspergillus niger ATCC16404.

[0082] The test method can refer to the Ministry of Health of the People's Republic of China, Disinfection Technical Specification Method:

[0083] The experimental results are shown in Table 2:

[0084] Antibacterial performance table 2:

[0085]

[0086]

[0087] It can be seen from Table 2 that the composition of the present invention can unexpectedly achieve synergy, unexpectedly obtain better antibacterial performance, and improve broad-spectrum antibacterial performance.

[0088] (2) Adhesion data

[0089] Adhesion force was studied using artificial saliva provided in different cases (experiments 1 to 3) of Example 1 and comparative examples 1 to 5.

[0090] Test methods for adhesion studies:

[0091] Test conditions: temperature 25±1℃.

[0092] Test tool: NDJ-8s viscometer.

[0093] Test process: Take a sample of the preparation and put it into the container of the viscometer. After it stabilizes, select the appropriate rotor and speed (rotor: No. 2, speed: 6 rpm). The range percentage reading should be between 10% and 100%, with 50% (50% ± 10%) being the most suitable.

[0094] The results of the experiment are as follows: Table 3

[0095] Table 3

[0096] Case Average viscosity (mPa·s) Example 1 - Experiment 1 506.5 Comparative Example 1 lacks CPC 375 Comparative Example 2 lacks HPMC 120 Comparative Example 3 lacks glycerol 603 Comparative Example 4: Chlorhexidine 413.5 Comparative Example 5: CMC 422

[0097] As shown in Table 3, the combination of components of the present invention can unexpectedly achieve synergy, so that the artificial saliva component can reach the use requirement of 500-550 mPa·s, making it meet the needs of oral use.

[0098] Part II: Preparation and property study of explosive beads

[0099] 2.1: Bead blasting case preparation case:

[0100] Example 2

[0101] Step 1): Preparation of artificial saliva

[0102] The ingredients were prepared and mixed according to the formula of Example 1 to prepare artificial saliva (the concentrations of HPMC and glycerol were used as orthogonal design factors (specific orthogonal assignments are shown in Table 2)), and then allowed to stand overnight in a refrigerator at 4±2°C to remove bubbles generated during the stirring process.

[0103] Step 2): Preparation of alginate solution

[0104] A total of 5 g of sodium alginate was dissolved in 1000 ml of distilled water and stirred using a magnetic stirrer until a clear solution appeared. Thereafter, it was filtered.

[0105] Place the mixture in a refrigerator at 4±2℃ overnight to remove bubbles generated during stirring.

[0106] Step 3): Preparation of stock solution.

[0107] Adding calcium chloride to the artificial saliva prepared in step 1) to control the concentration of calcium chloride in the mixed solution to be 4 wt.%;

[0108] Ultrasonic vibration was performed for 30 min and the mixture was placed in a 4°C refrigerator overnight.

[0109] Step 4): Preparation of artificial saliva LHBs

[0110] Step 4-a): The alginate solution (concentration 0.5 wt.%) was transferred into a transparent container and kept on a magnetic stirrer.

[0111] Step 4-b): Use a titrator to drop the stock solution of step 3) into the alginate bath for reverse spheroidization treatment, wherein the titration height is fixed at 8 cm and the speed is 1 drop / s, and the volume of the total stock solution is 0.5 times that of the alginate solution; the stirring speed during the dropwise addition is 500-600 rpm, and after the dropwise addition is completed, the droplets are allowed to harden for 10 minutes.

[0112] Step 4-c): Separate the beads from the alginate bath and rinse them alternately with anhydrous ethanol and deionized water to remove excess alginate.

[0113] Step 4-d): Separate the burst beads and place them in a 6% by mass calcium chloride solution for secondary curing for 10 minutes.

[0114] Step 4-e): Rinse alternately with anhydrous ethanol and deionized water to remove excess calcium salt and ethanol.

[0115] Step 4-f): The product obtained in step 4-e): is placed in glycerol for soaking and modification, then separated and rinsed with purified water, packaged, and stored for later use.

[0116] The concentrations of HPMC and glycerol in the artificial saliva and the time of glycerol soaking modification in step 4-f) were orthogonally studied. The orthogonal design is shown in Table 4:

[0117] Table 4:

[0118]

[0119] Preparation effect analysis:

[0120] The fourth group was selected as the optimal combination (final formula), at which the HPMC concentration was controlled at 1.75%, the glycerol concentration was 1%, and the glycerol soaking time was 15s. The advantages are as follows:

[0121] ① Diameter, weight (storage performance) (results see Figure 3 );

[0122] Store at 4±2°C, extract the gel beads after 1, 2, 3 and 4 weeks, and measure the sample diameter and weight twice (the results are taken as the arithmetic mean). Diameter / weight of group in week i and week j The mean difference is d ijk . Define the maximum mean difference of the kth group as d max =maxd ijk .

[0123] definition f e The smaller it is, the less variability there is in diameter / weight within the group.

[0124] The largest differences within the groups were found, and the fourth group had the smallest values ​​compared with the other groups, so the storage performance was the best at this time.

[0125] ②Sphericity (Outer roundness (results see Figure 4 )).

[0126] Calculate the maximum and minimum diameters of each group of gel beads after preparation.

[0127] The larger the SF, the greater the deformation; SF=0, standard sphere; SF<0.5, approximately spherical; SF=1, straight line.

[0128] Among the 9 groups of tests: the sphericity of the 4th group was the smallest and the deformation was the smallest.

[0129] ③ Load capacity LC = (W1-W2) / W1 (see the results Figure 5 ).

[0130] Fixing the concentration ratio of the fourth group helps to encapsulate more artificial saliva contents in the gel beads. This may be because HPMC and glycerol act as a fence during the formation of the gel beads, hindering the diffusion of the encapsulated artificial saliva contents into the surrounding solution.

[0131] Reason: During the formation of gel beads, HPMC and glycerol act as a fence, hindering the diffusion of the contents into the surrounding solution.

[0132] Average value: Group 4 = Group 5 = 0.946,

[0133] Standard deviation: Group 4 (0.012) < Group 5 (0.016).

[0134] Note: The load capacity of the fourth group is more stable. More artificial saliva contents are wrapped in the gel beads.

[0135] Table 5: Gel bead loading capacity values ​​in nine tests

[0136]

[0137] Note: The data in the table are mean ± standard deviation.

[0138] ④ Hardness-film diameter (results see Figure 6 )

[0139] Average hardness level (3.643): The fourth group (3.56) and the fifth group (3.667) are close.

[0140] Standard deviation: Group 4 (0.282) < Group 5 (0.611).

[0141] Note: The process of making gel beads in the fourth group is more stable;

[0142] Group 4 was at an upper-middle level among the 9 groups of experiments.

[0143] Note: If the membrane diameter is too thick, the taste is poor and it is difficult to bite; if it is too thin, it is easy to break and difficult to store. Therefore, the fourth group of experiments produced gel beads with better physical and chemical properties.

[0144] 2.2: Comparative case of bead explosion preparation:

[0145] Comparative Example A

[0146] Compared with the fourth group of experiments in Example 2, the only difference is that the forward spheroidization technology is used, that is, the alginate solution is added dropwise to the stock solution of artificial saliva and calcium salt. As a result, solid particles are obtained and the artificial saliva encapsulation fails.

[0147] Comparative Example B

[0148] Compared with the fourth group of experiments in Example 2, the only difference is that the second stage of calcium salt solidification treatment was not performed. As a result, the artificial saliva encapsulation failed and the bursting bead preparation failed.

[0149] Comparative Example C

[0150] Compared with the fourth group of experiments in Example 2, the only difference is that in step 4-f), the soaking modification time is: (a): soaking time is 0 min; (b): soaking time is 120 s.

[0151] The results show that group (a) is very brittle and easily broken; group (b) loses water and shrinks, indicating preparation failure.

[0152] Comparative Example D

[0153] Compared with the fourth group of experiments in Example 2, the only difference is that the artificial saliva components of the present invention are not used, and the artificial saliva components in Comparative Examples 1 to 5 are used for encapsulation.

[0154] The results of hardness and encapsulation efficiency of the materials prepared in Example 2 and Comparative Examples A to E are shown in Table 6

[0155] Table 6:

[0156]

[0157] It can be seen that the use of the synergistic artificial saliva of the present invention, in combination with the reverse spheroidization, two-stage coagulation, HPMC-glycerol joint immersion modification and joint control of the immersion time, can unexpectedly achieve synergy, can prepare and achieve encapsulation, and successfully prepare the popping beads. Not only that, the hardness of the popping beads can also be controlled before 3.5 to 3.7 suitable for the oral cavity, and it helps to improve the encapsulation rate.

[0158] Part III: Bead blasting performance data

[0159] Embodiment 3:

[0160] The artificial saliva bursting beads (HPMC concentration 1.75%, glycerol 1%; glycerol soaking time 15s) prepared in the fourth group of experiments in Example 2 were measured.

[0161] ①Stability test: accelerated method.

[0162] Before stabilization: average effective content 0.111%,

[0163] After being stabilized at 37°C for 90 days: the average effective content is 0.105%.

[0164] Description: The effective ingredient cetylpyridinium chloride in the prepared gel beads decreased by 5.41%, which is less than 10%. The storage period of the gel beads is 2 years.

[0165] ②Tolerance test: change the mobile phase ratio and column temperature while keeping other conditions unchanged.

[0166] The RSD of cetylpyridinium chloride was 0.122% (<2%).

[0167] Description: The preparation method has good tolerance and can withstand environmental changes.

[0168] ③Acute eye irritation test (Table 7):

[0169] No abnormality was found in the drugged eyes of all the test animals at 1h, 24h and 48h after administration; no abnormality was found in the drugged eyes of all the test animals until 72h. The average scores of the three animals at 24h, 48h and 72h were: conjunctival congestion = 0, conjunctival edema = 0, corneal damage = 0, iris damage = 0. There was no abnormality in the control eyes.

[0170] Table 7

[0171]

[0172] Description: The prepared gel beads are non-irritating

[0173] ④Acute oral toxicity test (Table 8): Oral administration at a dose of 5000 mg / kg body weight

[0174] In order to verify the safety of the preparation prescription, a control group was set according to the requirements of the acute oral toxicity test under the 2002 edition of the Technical Specifications for Disinfection.

[0175] 14d: No obvious poisoning symptoms or death were observed, and no abnormal body weight was found.

[0176] Observation period expired: gross anatomical examination and naked eye observation of the main organs revealed no abnormal changes

[0177] Table 8

[0178]

[0179] Description: The product obtained has a 0% mortality rate in the acute oral toxicity test on KM mice (greater than 5000 mg / kg body weight), which is practically non-toxic and meets the safety requirements of 2.3.13.1 of the 2002 edition of the Technical Specifications for Disinfection.

[0180] ⑤ Mouse bone marrow polychromatic erythrocyte micronucleus test (Table 9)

[0181] According to the 2002 edition of the Technical Specifications for Disinfection, the evaluation requirements for the micronucleus test of mouse bone marrow polychromatic erythrocytes stipulate that a control

[0182] According to the results of acute oral toxicity test: the LD50 of the test substance is greater than 5000mg / kg body weight. The dosage group is 5000mg / kg body weight as the high dose, and 2000mg / kg and 500mg / kg body weight as the medium and low doses.

[0183] Micronucleus rate: There was no significant difference between each dose group and the negative control group (P>0.05). The positive control group (cyclophosphamide) was significantly higher than the negative control group and each dose group, and the difference was significant (P<0.01).

[0184] PCE / NCE ratio: There was no significant difference among the negative control group, positive control group and the names of the tested samples in each dosage group.

[0185] Table 9

[0186]

[0187] Note: The sample was negative in the micronucleus test of mouse bone marrow polychromatic erythrocytes, which means it is practically non-toxic. It meets the safety requirements of 2.3.13.2 of the 2002 edition of the Technical Specifications for Disinfection.

[0188] Part 4: Determination of stability and safety of active ingredients in Explosive Beads:

[0189] Example 4

[0190] The active ingredients, safety, etc. of the explosive beads prepared in the fourth group of experiments in Example 2 were measured, and the results were:

[0191] ①Linear relationship:

[0192] In the prepared gel beads, it can be measured that when the concentration of cetylpyridinium chloride is 4.814-63.379 μg / mL, the peak area of ​​cetylpyridinium chloride has a good linear relationship, and the corresponding relationship between the peak area and the concentration is:

[0193] Y=13.90321X-3.54804, r=0.994

[0194] Description: The prepared product (Group 4) can be successfully prepared in a wide range before and after the effective concentration of the main drug CPC (cetylpyridinium chloride) (10μg / mL), and the performance is stable and recyclable.

[0195] ②Specificity experiment:

[0196] From the chromatogram of the reference substance, it can be seen that the retention time of cetylpyridinium chloride is about 12.939 min.

[0197] Note: Compared with the chromatograms of the standard and blank excipients, it can be seen that the standard and blank excipients do not interfere with the determination of the concentration of the main drug in our sample. Its specificity is good.

[0198] ③Sample content determination: external standard method

[0199] The content was calculated by peak area, and the results showed that the content of the active ingredient CPC in the three batches of gel beads was between 90.0% and 110.0%, which was in line with the requirements of the Disinfection Technical Specifications 2002 Edition.

[0200] ④ Microbial contamination test (Table 10): GB 15979-2002, ambient temperature 21.7°C, relative humidity 56% test conditions

[0201] Cetylpyridinium chloride gel beads are colorless and transparent.

[0202] The total bacterial colony count, total fungal colony count, coliform group, Staphylococcus aureus, Pseudomonas aeruginosa and beta-hemolytic Streptococcus were not detected.

[0203] Table 10

[0204]

[0205] Note: The test results of microbiological indicators meet the requirements of GB 15979-2002 "Hygiene Standards for Disposable Sanitary Products", and the microbiological indicators of the prepared product are qualified.

[0206] ⑤ Antibacterial test (Table 11)

[0207] According to the requirements of the standard, disinfection products need to undergo pharmacological verification of the finished products and evaluate their efficacy.

[0208] When the sample was exposed to water for 2 minutes, the average inhibition rates against Escherichia coli, Staphylococcus aureus, Candida albicans and Pseudomonas aeruginosa were 95.65%, 90.31%, 84.39% and 92.43%, respectively.

[0209] Table 11

[0210]

[0211] Description: The antibacterial effect of the prepared product meets the requirements of the appendix of GB 15979-2002 "Hygiene Standards for Disposable Sanitary Products" (when the antibacterial rate is 50-90%, it has an antibacterial effect; when the antibacterial rate is >90%, the product has a strong antibacterial effect).

Claims

1. A method for preparing artificial saliva bursting beads, characterized in that the steps include: Step (1): adding a raw material solution containing artificial saliva and a calcium source dropwise into a wall material solution, performing a reverse spheroidization treatment, and then separating to obtain primary popping beads; the wall material is a cross-linked product of alginic acid and its salts; Step (2): placing the primary bursting beads in a calcium source solution for secondary gelation, and then soaking and modifying them in glycerol to obtain the artificial saliva bursting beads; The time of immersion modification treatment is 12~18s; The artificial saliva has the following weight percentages of components: cetylpyridinium chloride 0.08-0.12wt.%, HPMC 1.6-1.8wt.%, glycerol content 1wt.%, electrolyte 0.2-0.25wt.%, sweetener 2.4-3.2wt.%, preservative: 0.1-0.2wt.%, pH 6-7; and the balance is water.

2. The method for preparing artificial saliva bursting beads according to claim 1, characterized in that, The electrolyte is at least one of potassium chloride and sodium chloride.

3. The method for preparing artificial saliva bursting beads as claimed in claim 1, characterized in that, The sweetener is at least one of xylitol and steviol glycoside.

4. The method for preparing artificial saliva bursting beads according to claim 1, characterized in that, The preservative is at least one of sorbic acid and sodium benzoate.

5. The method for preparing artificial saliva bursting beads according to claim 1, characterized in that, The pH adjuster is at least one of potassium dihydrogen phosphate, sodium hydroxide and potassium hydroxide.

6. The method for preparing artificial saliva bursting beads according to claim 1, characterized in that: The molecular weight of HPMC is 1~1.5 million.

7. The method for preparing artificial saliva bursting beads according to claim 1, characterized in that: The concentration of calcium source in the raw material solution is 4%~6%.

8. The method for preparing artificial saliva bursting beads according to claim 1, characterized in that: The alginate is at least one of sodium salt and potassium salt of alginic acid, and the concentration of the solute in the alginate solution is 0.25% to 1%.

9. The method for preparing artificial saliva bursting beads according to claim 1, characterized in that: In step (2), the calcium source solution is an aqueous solution of a water-soluble calcium salt.

10. The method for preparing artificial saliva bursting beads according to claim 9, characterized in that: In step (2), the calcium source solution is an aqueous solution of at least one of calcium chloride, calcium lactate, calcium nitrate, and calcium acetate, and the concentration of the solute in the calcium source solution is 4% to 6%.

11. The method for preparing artificial saliva bursting beads according to claim 1, characterized in that: The temperature of the secondary gelation process is 35~37℃; the time is 4~10min.

12. Artificial saliva bursting beads prepared by the preparation method according to any one of claims 1 to 11.

13. The artificial saliva bursting beads according to claim 12, characterized in that: The particle size of the artificial saliva bursting beads is 12-16 mm; the content of artificial saliva is 90-98 wt.%.

14. The artificial saliva bursting beads according to claim 12, characterized in that: The particle size of the artificial saliva bursting beads is 13-15 mm; the content of artificial saliva is 93-96 wt.%.

15. An application of artificial saliva bursting beads prepared by the preparation method according to any one of claims 1 to 11, characterized in that: The invention is used for preparing medicine for preventing and / or treating dry mouth.

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