CN118021746A

CN118021746A - Preparation method and application of orally disintegrating tablet for treating dental ulcer

Info

Publication number: CN118021746A
Application number: CN202410245653.XA
Authority: CN
Inventors: 尹忠; 赵万洲
Original assignee: Nanjing Merck Sitai Pharmaceutical Technology Co ltd
Current assignee: Nanjing Merck Sitai Pharmaceutical Technology Co ltd
Priority date: 2024-03-05
Filing date: 2024-03-05
Publication date: 2024-05-14
Anticipated expiration: 2044-03-05

Abstract

The invention relates to a preparation method and application of orally disintegrating tablets containing thymol and derivatives thereof. The orally disintegrating tablet contains thymol and thymol derivatives, vitamin E and chondroitin sulfate derivatives (CST), vitamin E and polyethylene glycol derivatives (TPGS), mannitol, gelatin and water. The orally disintegrating tablet can be formed by freeze-drying. According to the invention, on one hand, thymol is prepared into the derivative, so that the pungency of the thymol is effectively reduced, and the gasoline taste of the thymol is shielded; the thymol and the derivative of thymol are coated by vitamin E derivatives, so that the two components are effectively solubilized, and the bioadhesion of the micelle preparation is greatly enhanced; furthermore, by preparing the orally disintegrating tablet, the use compliance is improved, and the healing time of the canker sore surface is shortened. The orally disintegrating tablet can be used for treating canker sore, oral care and correcting related oral abnormality.

Description

Preparation method and application of orally disintegrating tablet for treating dental ulcer

Technical Field

The invention relates to the field of natural medicines, in particular to a preparation method and application of an orally disintegrating tablet for treating canker sore.

Background

The oral inflammatory diseases are common diseases and frequently-occurring diseases of stomatology, and have high clinical morbidity and recurrence rate, such as pericoronitis, gingivitis, periodontitis, oral mucosa diseases and the like. Oral ulcers (oral ulcers) are one of the clinically common oral mucosal diseases, and are manifested by locally apparent recurrent pain, and the ulcers locally have the characteristics of redness, swelling and pain, affecting the patient's diet, pronunciation, swallowing and emotion. The canker sore has an ambiguous etiology and is caused by various factors such as immunity, heredity, systemic diseases, infection, tumor radiotherapy and chemotherapy, environment and the like. At present, no specific medicine for treating dental ulcer exists, and an inducing elimination method and symptomatic treatment are adopted clinically. The existing products for relieving symptoms comprise stomatitis spray, dexamethasone acetate patch, guilin watermelon frost and the like, and the medicines have certain treatment effects, but have respective obvious defects.

Thymol (thymol, THY) with CAS number of 89-83-8 has antibacterial, antioxidant, antiinflammatory, etc. activities. THY has been attracting attention in inhibiting inflammation of oral mucosa due to the above-mentioned pharmacodynamic action. There are reports in China that the oral ulcer is proved to have better effect after 0.25% domiphen and THY are combined for treatment of chemotherapy [ Shao Fang. Chinese community physicians (medical profession) [ 2011, 13 (12), 161]. There are some traditional therapies for relief of oral ulcers using THY-containing essential oils abroad, but no systematic study has yet been made. In addition, foreign studies have demonstrated that THY has therapeutic effects on animals in ulcerative colitis models, which also laterally demonstrates that THY has relatively significant anti-inflammatory activity [Tahmasebi P,Abtahi Froushani SM,Afzale Ahangaran N.Thymol has beneficial effects on the experimental model of ulcerative colitis.Avicenna J Phytomed.2019Nov-Dec;9(6):538-550].

THY has problems in oral use, mainly poor water solubility, strong peppery taste, and strong gasoline taste. This has led to liquid formulations containing THY having to use lower concentrations of THY to reduce the pungency, use a concentration of organic solvents to solubilize, or add surfactants such as tween to solubilize, and use aromatic substances with strong odors (e.g., menthol, eucalyptus oil, etc.) to mask the taste of THY. Although this improves to some extent the patient's compliance with THY for the treatment of oral diseases, there is still a need for further optimisation: organic solvents, menthol, eucalyptus oil, etc. contained in the solution may further irritate the canker sore face, and the lower concentration of THY in the solution also limits the efficacy. In addition, the oral ulcer is a local mucosa tissue lesion, when THY is used for treating the local ulcer, the medicine concentration at the wound surface is increased as much as possible, the dilution and loss speed of the medicine by saliva is reduced, and the common solution preparation has higher fluidity, so that the aim is not easy to achieve.

Disclosure of Invention

The invention aims to overcome the defects of the existing THY liquid preparation, increase the solubility of THY, improve the curative effect of THY, increase the residence time of THY on the surface of an ulcer, reduce the pungency of THY and mask the bad taste of THY. On this basis, the THY orally disintegrating tablet formulation was prepared so that it could form a viscous system after rapid disintegration on the ulcer surface, so as to cover the ulcer surface and form a local high concentration drug environment thereon. The invention aims to develop a preparation method and application of an orally disintegrating tablet containing THY and derivatives thereof.

In order to solve the technical problems, the invention provides the following technical scheme: a preparation method of an orally disintegrating tablet for treating canker sore, the orally disintegrating tablet comprises the following components in parts by weight:

The preparation method of the orally disintegrating tablet comprises the following steps: mixing THY, THY-Tos, CST and TPGS in the weight ratio, dissolving with ethanol, evaporating to remove ethanol, adding water into the flask, shaking, fully hydrating the film on the wall of the flask, and performing ultrasound to obtain light blue opalescent solution; adding mannitol and gelatin into the solution, dissolving thoroughly, adding the solution into a bubble cap mould, and lyophilizing to obtain orally disintegrating tablet;

The sum of the mass of THY and THY-Tos in the orally disintegrating tablet is not more than one fifth of the sum of the mass of CST and TPGS;

The structural formula (I) of the thymol derivative THY-Tos is as follows:

the structural formula (II) of thymol THY is as follows:

The structural formula (III) of the derivative CST formed by the vitamin E and the chondroitin sulfate is as follows:

wherein n=a positive integer from 5 to 500, r=so ₃ Na;

The structural formula (IV) of the derivative TPGS formed by the vitamin E and the polyethylene glycol is as follows:

where n=a positive integer from 100 to 2000.

The preparation method of the orally disintegrating tablet comprises the following steps: mixing THY, THY-Tos, CST and TPGS according to the weight ratio, stirring and dissolving with 50ml absolute ethyl alcohol until the mixture is clear, and removing the organic solvent by rotary evaporation; 5mL of water was added to the flask, and the flask wall was thoroughly hydrated by shaking; ultrasonic treatment (100W, 5s/5s,10 min) to obtain 5mL light blue opalescent solution; taking 5mL of the micelle preparation, adding a certain amount of mannitol and gelatin, stirring and dissolving, adding 0.5mL of the solution into a bubble cap mould, pre-freezing the bubble cap filled with the solution at the temperature of minus 20 ℃ for 1h for shaping, covering with aluminum foil, pre-freezing at the temperature of minus 80 ℃ for 1h, taking out, punching holes on the aluminum foil, and freeze-drying for 24h at the temperature of minus 60 ℃ and 5Pa in a freeze dryer to obtain the freeze-dried orally disintegrating tablet.

Preferably, the orally disintegrating tablet comprises the following components in percentage by mass:

The preparation method of the orally disintegrating tablet comprises the following steps: THY, THY-Tos, CST and TPGS in the weight ratio are mixed, stirred and dissolved with 50ml absolute ethyl alcohol until the mixture is clear, and the organic solvent is removed by rotary evaporation. 5mL of water was added to the flask, and the flask wall was thoroughly hydrated by shaking; ultrasonic treatment (100W, 5s/5s,10 min) to obtain 5mL light blue opalescent solution. Taking 5mL of the micelle preparation, adding a certain amount of mannitol and gelatin, stirring and dissolving, adding 0.5mL of the solution into a bubble cap mould, pre-freezing the bubble cap filled with the solution at the temperature of minus 20 ℃ for 1h for shaping, covering with aluminum foil, pre-freezing at the temperature of minus 80 ℃ for 1h, taking out, punching holes on the aluminum foil, and freeze-drying for 24h at the temperature of minus 60 ℃ and 5Pa in a freeze dryer to obtain the freeze-dried orally disintegrating tablet.

In order to solve the technical problem of the invention, the invention provides another technical scheme that: the orally disintegrating tablets are useful for treating canker sores, or for medical devices or other oral care products.

The beneficial effects are that:

We have found, by accident, that in the early work, when THY and vitamin E are combined on the model of canker sore of animals, better therapeutic effect can be obtained than THY alone or vitamin E alone.

Based on the above findings, we have esterified THY with vitamin E succinate (vitamin E succinate, tos) to form THY derivatives (THY-Tos) starting from therapeutic drugs on the one hand. Surprisingly we found that the efficacy of THY-Tos on animal-based canker sore models was not significantly different compared to the combination of THY with vitamin E. We have also surprisingly found that the spicy and petrol taste of THY-Tos formed after esterification of THY with Tos is almost eliminated, which is beneficial to improving compliance in patients.

Considering that both THY and THY-Tos have low solubility, on the other hand, from the point of view of therapeutic drug carriers, we esterified chondroitin sulfate (chondroitin sulfate, CS) with Tos, which has bioadhesive properties, to form chondroitin sulfate derivatives (CST) which can spontaneously form micelle formulations at normal temperature. Further, we have unexpectedly found that mixing a water-soluble derivative of natural vitamin E, namely polyethylene glycol 1000 vitamin E succinate (D- α -tocopherol olyethyleneglycol 1000succinate, TPGS), with CST results in the formation of mixed micelles (CST/TPGS) in water at normal temperature, and the Critical Micelle Concentration (CMC) of the mixed micelles is lower than that of the critical micelles formed by CST alone, which makes the CST/TPGS mixed micelles more physically stable. We find that the CST/TPGS micelle has good entrapment and solubilization effects on THY and THY-Tos, and the formed THY micelle or THY-Tos mixture micelle can increase the solubility of THY or THY-Tos and has good stability. The drug-loaded CST/TPGS micelle preparation has better bioadhesion performance. When THY is entrapped in the mixed micelle carrier, THY-Tos is entrapped in the mixed micelle carrier, or a mixture of THY and THY-Tos is entrapped in the mixed micelle carrier, after the preparation is retained in the mucous membrane tissue for a period of time, more medicine is found to be retained on the mucous membrane, which is beneficial to the medicine to stay on the affected part of the canker sore for a longer time.

Furthermore, after a certain amount of mannitol as a lyoprotectant and gelatin as an adhesive are added into the micelle preparation, an orally disintegrating tablet can be formed after lyophilization. The amount of mannitol added in the micelle preparation is between 3.5% and 4.5% (w/v), and the amount of gelatin added in the micelle preparation is between 2% and 3% (w/v).

When the gelatin concentration is 2.0% -3.0%, the disintegration time of the orally disintegrating tablet increases with increasing gelatin concentration, but no longer exceeds 60s. When the gelatin concentration is higher than 3.0%, the disintegration time of the orally disintegrating tablets exceeds 60s. According to general requirements, the disintegration time of orally disintegrating tablets should not exceed 60s. The amount of gelatin to be added is preferably controlled to 2.0% to 3.0%.

At a mannitol concentration of 3.0%, the orally disintegrating tablet hardness cannot be measured, which results in an extremely easy breakage of the tablet during transportation or use. The hardness of the tablets increased significantly when mannitol concentration increased from 3.0% by 3.5% and 4.0%. When the mannitol concentration exceeds 4.0%, the hardness of the tablet is substantially stable. The amount of mannitol is not excessive considering that excessive oral administration of mannitol may cause hypertonic environment in the intestinal tract, possibly causing diarrhea. The amount of mannitol to be added is preferably controlled to 3.5 to 4.5%.

Preferably, the amount of mannitol added in the micelle preparation is 4% (w/v) and the amount of gelatin added in the micelle preparation is 2.5% (w/v). After the freeze-drying protective agent and the adhesive are added, the orally disintegrating tablet is obtained by freeze-drying molding, which is convenient to use, can also improve the medicine concentration at the wound surface, reduce the dilution and loss speed of the medicine by saliva and improve the healing speed of the ulcer surface.

The orally disintegrating tablet has the advantages compared with the liquid micelle preparation that: the wound dressing is convenient to use and good in wound retention; (2) The animal has better drug effect and faster healing speed on ulcer surface. The ulcer inhibition rate of the once-daily group of the orally disintegrating tablet prescription 5 can reach 92.7 percent in the fifth day, and exceeds the ulcer inhibition rate of all water solution type, micelle preparation type and positive medicine groups in the fifth day, which shows that the orally disintegrating tablet prescription has faster ulcer inhibition capability, and can be related to the formation of a viscous coating layer on the surface of an ulcer by the orally disintegrating tablet, and the local formation of high concentration of the medicine and difficult loss; the inhibition rate of the orally disintegrating tablet at the ninth day was 100%.

Drawings

FIG. 1 synthetic roadmap of THY-Tos

FIG. 2THY-Tos nuclear magnetic resonance hydrogen spectrum

FIG. 3 is a synthetic roadmap of CST

FIGS. 4a, b, c are nuclear magnetic resonance hydrogen spectra of CS, tos and CST, respectively

Appearance of orally disintegrating tablet prepared in FIG. 5

Detailed Description

The invention will now be described in further detail with reference to the accompanying drawings. The following description of the preferred embodiments of the present invention is provided for the purpose of illustration and explanation only and is not intended to limit the present invention.

Example 1: aqueous solution of THY

THY has poor solubility in water and is reported to have a water solubility of not more than 1mg/ml at 25 ℃ (https:// echa. Europa. Eu/registration-dossier/-/registered-dossier/11030/4/1). Aqueous solutions of low, medium, and high concentrations of THY were prepared at ambient temperature for subsequent comparative evaluation with micelle formulations. The preparation method comprises the following steps: adding a proper amount of THY raw material medicine into a beaker, adding 50mL of water, and magnetically stirring for 72h at normal temperature to respectively form THY aqueous solutions with the concentration of 0.1 mg/mL, 0.5 mg/mL and 0.9 mg/mL.

TABLE 1 aqueous solution of THY

Solution numbering	Solution concentration (mg/ml)
		1 (Low concentration)	0.1
2 (Medium concentration)	0.5
		3 (High concentration)	0.9

Example 2: TPGS micelle preparation of THY

THY is poorly water soluble and can be solubilized by the addition of organic solvents or by the addition of surfactants. The surfactant TPGS was selected for solubilization. The preparation method comprises the following steps: 40mg of THY is weighed and placed in a beaker, a certain amount of water and TPGS are added into the beaker, and the mixture is magnetically stirred for 24 hours at normal temperature to obtain the THY. Wherein, the mass ratio of THY to TPGS in the micelle preparation 2 to the micelle preparation 8 is 2: 1. 1: 1. 1:2.5, 1: 5. 1: 7. 1: 10. 1:15 and 1:20. the above micelle preparation state was visually observed.

TABLE 2 TPGS micelle preparation of THY

This study showed that when the apparent concentration of THY was 2mg/ml, the formulation exhibited a cloudy state and a precipitate without the addition of TPGS, indicating poor water solubility of THY. After the TPGS is gradually added, the preparation system changes from milky turbid to milky turbid until becoming semitransparent and transparent, which shows that the TPGS has better solubilization effect on THY. In particular when the mass ratio of THY to TPGS is 1:7 or TPGS with larger ratio has better solubilization effect and stable micelle system.

Example 3: THY and Tos react through esterification to form THY-Tos

Tos (100 mg) was dissolved in 5mL of Dichloromethane (DCM), dicyclohexylcarbodiimide (DCC, 46.65 mg) and 4-dimethylaminopyridine (DMAP, 2.3 mg) were added thereto and stirred at room temperature for 2h to give solution A. Solution A was added dropwise to 2mL of DCM solution containing THY (42.45 mg), reacted at 40℃for 36h, the reaction solution was dried under reduced pressure, and the resulting residue was isolated by preparative silica gel plate chromatography to give THY-Tos 63mg as a colorless oil in 50% yield. The synthesis of THY-Tos is shown in FIG. 1. The nuclear magnetic resonance hydrogen spectrum (¹ H-NMR) of THY-Tos is shown in FIG. 2 .¹H NMR(400MHz,Chloroform-d)δ7.21(d,J＝7.9Hz,1H,H-3'),7.05(dd,J＝8.0,1.8Hz,1H,H-4'),6.83(d,J＝1.7Hz,1H,H-6'),3.07(s,4H,H-11,H-12),3.00(m,1H,H-7'),2.61(t,J＝6.8Hz,2H,H-7),2.32(s,3H,Ar-CH₃),2.11(s,3H,Ar-CH₃),2.04(s,3H,Ar-CH₃),2.00(s,3H,Ar-CH₃),1.89–1.73(m,2H,H-6),1.64–1.47(m,4H),1.46–1.25(m,14H),1.19(d,J＝6.9Hz,6H,7'-CH₃,7'-CH₃),1.17–1.02(m,6H),0.93–0.84(m,12H,Aliphatic-CH₃).

Example 4: TPGS micelle preparation of THY-Tos

The preparation method of the TPGS micelle preparation of THY-Tos was similar to that of the THY micelle preparation of example 2. 40mg of THY-Tos was weighed into a beaker, to which a certain amount of water and TPGS were added; wherein, the mass ratio of THY-Tos to TPGS in the preparation 2 to the preparation 5 is 1:1. 1: 7. 1:10 and 1:15. the micelle preparation is magnetically stirred for 24 hours, and the preparation is obtained. The micelle preparation state was visually observed.

TABLE 3 TPGS micelle preparation of THY-Tos

This study shows that THY-Tos has poor solubility, and TPGS is effective in increasing the solubility of THY-Tos, especially when the mass ratio of THY-Tos to TPGS is 1:7 or TPGS with larger ratio has better solubilization effect and stable micelle system.

Example 5: CS and Tos form CST by esterification

Tos (796.17 mg), 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI, 0.5751 g), N-hydroxysuccinimide (NHS, 0.3453 g) and DMAP (85.52 mg) were added to N, N-dimethylformamide (DMF, 12 mL) and stirred at 30 ℃ for 3h to activate the carboxyl group of Tos. CS (530.78 mg) in DMF (6 mL) was slowly added to the activated Tos solution. The reaction was stirred at 30℃for 48h. The solution was precipitated with ice-cold acetone to remove excess Tos. Dissolving the precipitate in pure water, dialyzing with dialysis membrane (molecular weight cut-off: 3.5 kDa), removing catalyst and solvent, and lyophilizing. The synthetic route for CST is shown in FIG. 3. The nuclear magnetic resonance hydrogen spectra of CS, tos and CST are shown in FIG. 4.

As can be seen from ¹ H-NMR, the hydrogen spectrum of the dialysis-purified CST shows a characteristic peak for Tos, indicating that Tos is attached to CS by esterification.

Example 6: CST/TPGS mixed preparation for preparing THY and/or THY-Tos

Appropriate amount of THY, THY-Tos, or THY in combination with THY-Tos, and appropriate amount of CST and TPGS were mixed, dissolved with 50ml of absolute ethanol under stirring until clear, and the organic solvent was removed by rotary evaporation. To the flask was added 5mL of water, and the flask wall was thoroughly hydrated by shaking. Ultrasonic treatment (100W, 5s/5s,10 min) to obtain 5mL light blue opalescent solution.

TABLE 4 CST/TPGS micelle solution of THY and/or THY-Tos

This study demonstrates that CST and TPGS can form mixed micelles and successfully solubilize THY and/or THY-Tos. In addition, the mixed micelle formed by CST and TPGS has better solubilization capacity compared with the TPGS micelle. This is shown in: when the ratio of the total mass of THY and THY-Tos to the total mass of CST and TPGS is 1:7, forming a system which is a light blue opalescent clear micelle solution system; from examples 2 and 4, it is understood that the ratio of the mass of THY or THY-Tos to the mass of TPGS is 1:7, the system is not completely clear and shows semitransparent characteristics. Further, according to the experiment of the solution 6 of this example, we have also surprisingly found that when the ratio of the total mass of THY and THY-Tos to the total mass of CST and TPGS is 1:5, a light blue opalescent clear micellar solution system was still formed, which is sufficient to demonstrate its greater solubilising power.

Example 7: comparison of pungency degree

6 Volunteers with good physical health, olfactory and gustatory sensitivity (ratio of male to female 1:1) were selected, and part of the formulations in examples 1, 2, 4 and 6 were tasted and scored, each volunteer was scored for micelle solution alone, and the evaluation criteria are shown in Table 5. The results of scoring the mouthfeel of all volunteers were recorded, and the statistical average was taken as the sensory evaluation result.

Table 5 taste scoring criteria

Table 6 piquancy degree test of each solution and formulation

This study demonstrates that THY has a relatively strong pungent taste when dissolved in water as a pure solution, either at low concentration (0.1 mg/ml) or at high concentration (0.9 mg/ml). Particularly high concentrations of THY in water, all volunteers considered to be unacceptably hot. After THY is prepared into TPGS micelle solution, the pungency is reduced, especially when the mass ratio of THY to TPGS is 1:10, or a larger TPGS duty cycle, the reduction in peppery taste is more pronounced, which is seen in all volunteers.

On the other hand, esterification of THY with Tos resulted in the formation of THY-Tos with all of its pungency removed. It should be noted that when THY-Tos is absorbed into the oral mucosa, the abundance of hydrolytic enzymes present in the oral tissue theoretically hydrolyzes the ester bonds formed by the association of THY and Tos, releasing THY and Tos; furthermore, tos can be further hydrolyzed to form vitamin E, so that THY and vitamin E synergistically interfere with canker sore, and the wound healing is promoted.

The THY-Tos needs time for hydrolysis, and the THY-Tos micelle is used for wound surface only, and can release THY in the internal tissue of the wound surface for a long time in theory, but the THY is helpful to take effect more quickly if the THY is contained in the preparation at the same time when the medicine is just started. Therefore, we co-load THY and THY-Tos in CST/TPGS mixed micelles and confirmed that the degree of pungency was significantly reduced in the volunteer's attempts at the degree of pungency of each mixed micelle. The reason for the reduced pungency is, on the one hand, because the micelles contain both non-pungent THY-Tos and small concentrations of THY, and, on the other hand, because of the effective encapsulation of THY and THY-Tos by CST/TPGS.

Example 8: bioadhesion contrast

The bioadhesion of the formulations in some of the examples was evaluated using the rat in vitro intestinal perfusion method. Sprague Dawley rats (220 g.+ -.20 g) were divided into 7 groups of 10 animals each. After the sacrifice, the colon was taken out, sheared down along the cecum, the contents were rinsed off with pH 7.4 phosphate buffer and attached to an inclined fixed tube, 10ml of each of the formulations in the examples in the table below was dropped from the upper port of the inclined tube, the eluted formulation was taken out from the lower port of the inclined tube with a beaker, and the slow dropping was performed for 5 minutes. After the dripping is completed, 20ml of pure water is dripped from the upper opening of the inclined tube again, the same beaker is used for receiving the eluent, and the eluent is slowly dripped for 10min. Methanol was added to the eluted solution to a volume of 50ml, shaken for 24 hours, centrifuged, and the supernatant was taken, quantified for THY by HPLC-UV, and bioadhesion was calculated according to the following formula.

Bioadhesion = (n ₁–n₂)/n₁ x 100%

Where n ₁ is the THY content in 10ml of the test sample before priming and n ₂ is the THY content in the eluted solution.

Table 7 bioadhesion test of each formulation

This example attempts to evaluate bioadhesion of each formulation with intestinal mucosa instead of buccal mucosa. It is clear that if a formulation has good bioadhesion, it is retained by the mucosa in a high proportion when it passes through the rat intestinal tract. Both solutions in example 1 were simple aqueous solutions of THY, with no significant bioadhesion. Both solutions in example 2 are TPGS micelle formulations of THY, which exhibit some bioadhesion due to the presence of polyethylene glycol (PEG) segments in the TPGS. Several of the solutions in example 6 were mixed micelle solutions of THY, THY-Tos, exhibiting significant bioadhesion; in particular, when the ratio of the total mass of THY and THY-Tos to the total mass of CST and TPGS is 1:15 (example 6 solution 8) and 1:20 (example 6 solution 9) all had bioadhesions exceeding 70%. The bioadhesion of the CST/TPGS mixed micelle preparation is obviously superior to that of a micelle preparation formed by TPGS alone.

Example 9: increasing the viscosity of the formulation

A thickening formulation is prepared. The preparation method comprises the following steps: the materials were weighed according to the following table, placed in a beaker, and water was added to the full volume. Magnetically stirring for 24h to obtain the final product. Each formulation viscosity was measured using an NDJ-5S rotary viscometer (Shanghai Changji geological instruments Co., ltd.) with a number 2 rotor.

TABLE 8 viscosity test of thickened formulations

As can be seen from the above table, without the addition of sodium carboxymethylcellulose, the viscosity of the formulation is very low, which gives the following results: when the preparation is applied to the affected part, the loss of the liquid medicine is easily caused due to the high fluidity of the preparation itself. After sodium carboxymethyl cellulose is added into the preparation system, the viscosity of the preparation increases along with the increase of the concentration of sodium carboxymethyl cellulose. When the preparation with certain viscosity is used on the affected part of the oral cavity, the fluidity of the liquid medicine is weakened, which is beneficial to improving the compliance of patients in use.

Example 10: preparation of orally disintegrating tablets

The following three micelle formulations were first prepared:

(I) TPGS micelle formulation group 0.2% thy: formulated as shown in example 2, with a THY concentration of 2mg/ml in the formulation (example 2 solution 8);

(II) CST/TPGS mixed micelle formulation group of 0.2% thy: formulated as shown in example 6, the concentration of THY in the formulation was 2mg/ml (example 6 formulation 1).

(III) CST/TPGS mixed micelle preparation group of 0.1% THY and 0.1% THY-Tos: formulated as shown in example 6, the concentrations of THY and THY-Tos in the formulation were 1mg/ml (example 6 formulation 8).

Taking 10ml of each micelle preparation, respectively adding mannitol with different amounts to ensure that the concentration of mannitol in the liquid micelle preparation is 3.0%, 3.5%, 4.0%, 4.5%, 5.0% and 5.5% (w/v), stirring and dissolving, and then adding gelatin to ensure that the concentration of gelatin in the liquid micelle preparation is 2.5% (w/v). Adding 0.5ml of the solution into a bubble cap mould, pre-freezing the bubble cap filled with the solution at the temperature of minus 20 ℃ for 1 hour for shaping, covering with aluminum foil, pre-freezing at the temperature of minus 80 ℃ for 1 hour, taking out, punching holes on the aluminum foil, and freeze-drying for 24 hours at the temperature of minus 60 ℃ under the condition of 5Pa in a freeze dryer to obtain the freeze-dried orally disintegrating tablet. The hardness of each group of orally disintegrating tablets was measured.

Table 9 hardness (N) of orally disintegrating tablets of THY micelle preparation

From the above results, it was found that when the mannitol concentration was 3.0%, the orally disintegrating tablet hardness could not be measured, which resulted in an extremely easy breakage of the tablet during transportation or use. The hardness of the tablets increased significantly when mannitol concentration increased from 3.0% by 3.5% and 4.0%. When the mannitol concentration exceeds 4.0%, the hardness of the tablet is substantially stable. The amount of mannitol is not excessive considering that excessive oral administration of mannitol may cause hypertonic environment in the intestinal tract, possibly causing diarrhea. The amount of mannitol to be added is preferably controlled to 3.5 to 4.5%.

Example 11: preparation of orally disintegrating tablets

The following three micelle formulations were first prepared:

Taking 10ml of each micelle preparation, respectively adding mannitol to ensure that the concentration of mannitol in the liquid micelle preparation is 4.0% (w/v), stirring and dissolving, and then adding gelatin to ensure that the concentration of gelatin in the liquid micelle preparation is 2%, 2.5%, 3%, 3.5% and 4% (w/v). Adding 0.5ml of the solution into a bubble cap mould, pre-freezing the bubble cap filled with the solution at the temperature of minus 20 ℃ for 1 hour for shaping, covering with aluminum foil, pre-freezing at the temperature of minus 80 ℃ for 1 hour, taking out, punching holes on the aluminum foil, and freeze-drying for 24 hours at the temperature of minus 60 ℃ under the condition of 5Pa in a freeze dryer to obtain the freeze-dried orally disintegrating tablet. The disintegration time of each group of orally disintegrating tablets was measured.

Table 10 disintegration time(s) of orally disintegrating tablets of the THY micelle preparation

From the above results, it was found that when the gelatin concentration was 2.0% to 3.0%, the disintegration time of the orally disintegrating tablet increased with increasing gelatin concentration, but none exceeded 60s. When the gelatin concentration is higher than 3.0%, the disintegration time of the orally disintegrating tablets exceeds 60s. According to general requirements, the disintegration time of orally disintegrating tablets should not exceed 60s. The amount of gelatin to be added is preferably controlled to 2.0% to 3.0%.

Example 12: preparation of orally disintegrating tablets and volunteer attempts

Firstly, preparing a TPGS micelle preparation group with the THY of 0.2 percent,

Orally disintegrating tablets were prepared according to the compositions of the substances in the following table, see table 10.

Table 11 preparation of orally disintegrating tablets

The preparation method of the orally disintegrating tablet corresponding to each prescription comprises the following steps: mixing THY, THY-Tos, CST and TPGS according to the weight ratio, stirring and dissolving with 50ml absolute ethyl alcohol until the mixture is clear, and removing the organic solvent by rotary evaporation; 5mL of water was added to the flask, and the flask wall was thoroughly hydrated by shaking; ultrasonic treatment (100W, 5s/5s,10 min) to obtain 5mL light blue opalescent solution; taking 5mL of the solution, adding a proper amount of mannitol and gelatin, stirring and dissolving, adding 0.5mL of the solution into a bubble cap mould, pre-freezing the bubble cap filled with the solution at the temperature of minus 20 ℃ for 1h for shaping, covering with aluminum foil, pre-freezing at the temperature of minus 80 ℃ for 1h, taking out, punching holes on the aluminum foil, and freeze-drying for 24h at the temperature of minus 60 ℃ and 5Pa in a freeze dryer to obtain the freeze-dried orally disintegrating tablet. As shown in fig. 5.

The orally disintegrating tablets of each group prepared in example 12 were taken by 8 volunteers, and preliminary evaluations were made on the convenience of use, the pungency and the disintegration situation of the orally disintegrating tablets. All volunteers feed back the disintegrating tablets of each group, so that the disintegrating tablets are convenient to use and have no obvious pungency, and can be completely disintegrated in the oral cavity within 1 minute.

In addition, all volunteers fed back that each group of orally disintegrating tablets was able to form a layer of viscous substance on the canker face after disintegration of the canker face, which was much lower in fluidity than the solution formulation and was able to remain on the canker face for a longer period of time.

Example 13: pharmacodynamic study

(1) Medicine preparation method

(I) 0.05% THY aqueous solution group: formulated as shown in example 1 with a THY concentration of 0.5mg/ml in the final solution (example 1 solution 2);

(II) 0.05% thy and 0.05% vitamin E in water: the preparation method comprises the following steps: adding appropriate amount of THY and vitamin E into beaker, adding 50mL of water (containing 10% and 5% ethanol and PEG400 by volume fraction respectively), magnetically stirring at normal temperature for 72h, and keeping THY and vitamin E concentration in the final solution at 0.5mg/mL.

(III) group of 0.05% THY-Tos aqueous solutions: the preparation method comprises the following steps: an appropriate amount of THY-Tos was added to the beaker, 50mL of water (containing 10% and 5% ethanol and PEG400 by volume fraction, respectively) was added, and magnetically stirred at room temperature for 72 hours, the concentration of THY-Tos in the final solution being 0.5mg/mL.

(IV) TPGS micelle formulation group 0.2% thy: formulated as shown in example 2, with a THY concentration of 2mg/ml in the formulation (example 2 solution 8);

(V) thickened 0.2% thy TPGS micelle formulation group: formulated as shown in example 9 with a THY concentration of 2mg/ml (example 9 solution 2);

(VI) CST/TPGS mixed micelle formulation group of 0.2% thy: formulated as shown in example 6, the concentration of THY in the formulation was 2mg/ml (example 6 formulation 1).

(VII) CST/TPGS mixed micelle preparation group of 0.2% THY-Tos: formulated as shown in example 6, the concentration of THY in the formulation was 2mg/ml (example 6 formulation 5).

(VIII) CST/TPGS mixed micelle preparation group of 0.1% THY and 0.1% THY-Tos: formulated as shown in example 6, the concentrations of THY and THY-Tos in the formulation were 1mg/ml (example 6 formulation 8).

(IX) orally disintegrating tablets prescription 5 groups: prepared according to prescription 5 of example 12.

(2) Animals

Source, germ line, strain: SPF-grade SD rats, offered by Peking Vitrendy laboratory animal technologies Inc. Experimental animal production license: SCXK (su) 2019-0001; experimental animal use license: SYXK (su) 2023-0031; animal body weight: 180-220g; sex of animals: the male and female parts are half.

(3) Experimental instrument

DK-8D constant temperature water bath, shanghai Jing HongJi laboratory Equipment Co., ltd; sartorius electronic balance, certolisco instruments (beijing) limited.

(4) Experimental method

After the rats are adaptively fed for 3 days, the rats are fixed on an operation table after anesthesia, SD rat labial mucosa is dried by using a sterile cotton ball, and the sterile cotton ball is used for moisture isolation, naOH crystals with the thickness of 3mm multiplied by 3mm are placed on the labial mucosa, after the NaOH crystals are maintained for 30 seconds, the treated part is dipped by using a sterile cotton ball containing physiological saline, and the residual NaOH is removed; after molding, the compositions were randomly divided into a normal group, a model group, a once-per-day 0.05% THY aqueous solution, a three-per-day 0.05% THY aqueous solution, a once-per-day 0.05% THY and 0.05% vitamin E aqueous solution, a once-per-day 0.05% THY-Tos aqueous solution, a once-per-day 0.2% THY micelle preparation, a three-per-day 0.2% THY micelle preparation, a once-per-day thickened TPGS micelle preparation of 0.2% THY, a twice-per-day thickened TPGS micelle preparation of 0.2% THY, a mixed micelle preparation of CST/TPGS of 0.2% THY-Tos, a once-per-day CST/TPGS of 0.1% THY and 0.1% THY-Tos, a once-per-day oral tablet prescription, a once-per-day 5-per-day, a once-per-day daily-disintegrating watermelon in a three-day set of a Guartum. The Guilin melon cream group is externally applied to the wound surface for 9 days, and the rest of the administration groups are instilled for 9 times and 9 days. The control group and the model group were respectively administered with physiological saline by gastric lavage.

The oral ulcer area was macroscopically assessed using an area method, and the maximum transverse and longitudinal diameters of the ulcer were measured with vernier calipers, labeled d ₁ and d ₂, respectively, with an ulcer area s=pi×d ₁×d₂ ×1/4 (pi=3.14).

Ulcer area inhibition = (1-mean ulcer area in dosing group/mean ulcer area in model group) ×100%

(5) Statistical method

Analysis was performed using t-test, and normal distribution quantitative data was obtained as mean ± standard deviationAnd the method is characterized in that the single-factor analysis of variance and the LSD method are adopted for pairwise comparison, and Wilcoxon rank sum test is adopted when the analysis of variance condition is not met. Taking alpha=0.05, P <0.05 is statistically significant for the difference, P <0.01 is statistically significant for the difference, and P >0.05 is statistically non-significant for the difference.

(6) Effects of each pharmaceutical formulation on oral mucosa of rats of each group

The ulcer areas of each group are shown in Table 12.

TABLE 12 Effect of THY formulations on the ulcerated area and ulceration inhibition rate of rats with canker sore N=10, ulcer area unit: mm ²)/>

^*P<0.05,^** P <0.01 compared to model set

Summarizing the study results: (1) Compared with the model group, the ulcer inhibition rate of the group with 0.05% THY water solution is 69.8% on the ninth day of administration; the 0.05% THY in water showed 87.7% inhibition of ulcers in the three-day group on day nine of dosing, and a significant reduction in the area of ulcers compared to the model group (P < 0.01). (2) The ulcer inhibition rate of the 0.05% THY and 0.05% vitamin E water solution once daily group and the 0.05% THY-Tos water solution once daily group exceeds 80% on the ninth day of administration, and the effect is superior to that of the single 0.05% THY water solution once daily group, which shows that THY and vitamin E can act synergistically on dental ulcer; (3) The 0.2% thy micelle formulation had an ulcer inhibition of 85.4% once daily for the ninth day of dosing; the 0.2% thy micelle formulation had 100% inhibition of ulcers three times daily in the ninth day of dosing. The therapeutic effect of the 0.2% THY micelle preparation in the three-times-daily group at the ninth day of administration is better than that of the 0.05% THY aqueous solution in the three-times-daily group at the ninth day of administration; (4) Thickening 0.2% THY micelle solution can reach 100% of ulcer inhibition rate in the second daily group on the ninth day of administration, which indicates that adding the thickener in the preparation is helpful for improving the treatment effect. (5) The treatment effects of the CST/TPGS mixed micelle preparation of 0.2% THY and the CST/TPGS mixed micelle preparation of 0.2% THY-Tos are good, the ulcer inhibition rate of the former is close to 94%, the ulcer inhibition rate of the latter is 100%, and the high ulcer inhibition rate of the latter and the former show that the mixed micelle can improve the curative effect of the medicament by increasing the bioadhesion of the preparation on the ulcer surface; (6) The once daily co-formulation of the CST/TPGS mixed micelle of 0.1% THY and 0.1% THY-Tos achieved 100% inhibition of ulcers on the ninth day of administration, and we note that the once daily co-formulation of the CST/TPGS mixed micelle of 0.1% THY and 0.1% THY-Tos showed a near 80% inhibition of ulcers on the third day of administration, whereas at the same time the once daily co-formulation of the CST/TPGS mixed micelle of 0.2% THY-Tos showed a near 65% inhibition of ulcers on the initial stages of administration when THY and THY-Tos co-exist, which showed a more rapid inhibition of ulcers; the inhibition rate of ulcers on the fifth day was comparable for the once daily group of the 0.1% THY and 0.1% THY-Tos CST/TPGS mixed micelle formulation and the once daily group of the 0.2% THY-Tos CST/TPGS mixed micelle formulation; (7) The ulcer inhibition rate of the once-daily group of the orally disintegrating tablet prescription 5 can reach 92.7 percent in the fifth day, and exceeds the ulcer inhibition rate of all water solution type, micelle preparation type and positive medicine groups in the fifth day, which shows that the orally disintegrating tablet prescription has faster ulcer inhibition capability, and can be related to the formation of a viscous coating layer on the surface of an ulcer by the orally disintegrating tablet, and the local formation of high concentration of the medicine and difficult loss; the inhibition rate of the orally disintegrating tablet on the ninth day is 100%; (8) The inhibition rate of ulcer is only 88.4% when the drug is used with Guilin watermelon frost as a positive drug group for three times daily on the ninth day.

Claims

1. A method for preparing orally disintegrating tablets for treating canker sore, which is characterized in that: the orally disintegrating tablet comprises the following components in parts by weight:

The structural formula (I) of the thymol derivative THY-Tos is as follows:

the structural formula (II) of thymol THY is as follows:

wherein n=a positive integer from 5 to 500, r=so ₃ Na;

where n=a positive integer from 100 to 2000.

2. The method for preparing orally disintegrating tablet for treating canker sore according to claim 1, wherein: the orally disintegrating tablet comprises the following components in percentage by mass:

3. The method for preparing orally disintegrating tablet for treating canker sore according to claim 2, wherein: the orally disintegrating tablet comprises the following components in percentage by mass:

4. a method for preparing orally disintegrating tablets for treating canker sores according to claim 3, characterized in that: the orally disintegrating tablet comprises the following components in percentage by mass:

5. Use of an orally disintegrating tablet for treating canker sores prepared according to any one of claims 1 to 4, characterized in that: the orally disintegrating tablet is used for preparing medicines for treating dental ulcers or used as medical equipment or other oral care products.