CN114983982B

CN114983982B - Sustained-release type oral ulcer hydrogel ulcer patch

Info

Publication number: CN114983982B
Application number: CN202210598861.9A
Authority: CN
Inventors: 高长有; 汪凯; 董晓飞
Original assignee: Shaoxing Research Institute Of Zhejiang University; Zhejiang University ZJU
Current assignee: Shaoxing Research Institute Of Zhejiang University; Zhejiang University ZJU
Priority date: 2022-05-30
Filing date: 2022-05-30
Publication date: 2023-09-29
Anticipated expiration: 2042-05-30
Also published as: CN114983982A

Abstract

The invention discloses a slow-release type dental ulcer patch and a preparation method thereof, wherein the dental ulcer patch is applied by hydrogel containing hyperbranched polylysine (HBPL), wherein the content of the HBPL is 100 mug-200 mg, the hydrogel is hydrogel based on acrylic acid, acrylamide and derivatives thereof and polysaccharide hydrogel, the HBPL is loaded in the hydrogel through hydrogen bond action or Schiff base action, the hydrogen bond action or the Schiff base action damage is caused under the acidic environment of a dental ulcer wound, the slow release of the HBPL at the wound is realized, and the released HBPL can kill bacteria on the surface of the wound and simultaneously has the effect of eliminating inflammation. The slow-release type dental ulcer patch can accelerate the healing of dental ulcer wound surfaces, has no toxic or side effect, can not cause bacterial drug resistance after long-term use, and has obvious advantages compared with hormone drugs and immune promoters.

Description

Sustained-release type oral ulcer hydrogel ulcer patch

Technical Field

The invention belongs to the field of biomedical polymer materials, and particularly relates to a slow-release type canker sore hydrogel ulcer patch.

Background

Canker sore is a common oral mucosa disease, also called aphtha, and is obvious in pain during attack, and can be self-healed without treatment. Recurrent attacks, known as recurrent oral ulcers, have a great impact on the work and life of the patient. Clinically, ulcers are classified into three types, light, heavy and herpes-like, according to the size, number and duration of the ulcers. The light to moderate dental ulcer brings discomfort to the daily life of people, the eating of people can be influenced by the severe dental ulcer, and meanwhile, the dental ulcer has the characteristics of high morbidity and easy recurrence after healing, and the normal life of people is seriously influenced.

The current medicines for treating canker sore mainly comprise: immunopotentiators or hormonal drugs. The medicine has wide application in foreign countries, can effectively treat some ulcers which are deep and can not be healed for a long time, and can inhibit the recurrence of the ulcers for 3-6 months. However, the long-term use of hormones can produce certain serious adverse side effects. The immunopotentiator can also reduce the recurrence of ulcer, such as transfer factor, thymus peptide, etc., but the curative effect of the medicine is possibly reduced after long-term use. Up to now, no safe and effective therapeutic drug for treating the dental ulcer diseases exists, so that the development of a novel therapeutic drug for treating the dental ulcer with high curative effect is a difficult problem to be solved at present.

The film agent is a film-shaped preparation prepared by processing the medicine and a proper film-forming material, and is used for oral administration or external application of mucous membrane. In recent years, mucoadhesive formulations developed by bioadhesion technology at home and abroad have been rapidly developed. The film agent can be tightly adhered to the mucous membrane, so that the compactness and the persistence of the contact between the medicine and the mucous membrane are enhanced, the slow release effect of the medicine is achieved, and the medicine effect is greatly improved. For the special physiological environment of the oral cavity, the oral film can be attached to the surface of a mucous membrane, so that the ulcer can be mechanically protected, the medicine can directly reach the affected part, and the medicine can be released in a positioning way, so that the treatment effect can be greatly improved.

Chinese patent application 201410261996.1 discloses a patch for treating oral ulcer and a preparation method thereof, wherein the patch is a nanofiber non-woven fabric, the non-woven fabric is composed of core-shell structure nanofibers, a core-shell structure nanofiber shell layer is a polyethylene oxide shell layer, and a core-shell structure nanofiber core layer is a mixture of dexamethasone, vitamin C and vitamin B2. The prepared film can ensure enough medicine storage and release area, and the dissolution rate of the used polyethylene oxide in the oral cavity is moderate, so that the ulcer can be continuously kept at the effective medicine concentration, and the curing of the dental ulcer is facilitated. However, dexamethasone is used as a hormone drug, and long-term use can produce some serious adverse side effects.

Chinese patent CN103142562B discloses a slow release film for treating stomatocace, which has a three-layer structure, and the upper layer is a slow release coating contacting with the stomatocace surface; the lower layer is a polyacrylic acid film with a protective effect; the middle layer is composed of bacterial cellulose/drug-carrying substance composite membrane, and the drug-carrying substance is a mixture of one or more of borneol, artificial bezoar, dexamethasone acetate and vitamin B2, glycerol, dextrin, carbomer, polyethylene glycol and sodium carboxymethyl cellulose. The prepared slow-release film has good adhesiveness, air permeability and biocompatibility, large drug-loading rate and long local action time, and can relieve pain, promote wound healing and prevent wound infection. Dexamethasone acetate, which is also used, is a hormonal drug, and can produce certain serious adverse side effects after long-term application.

There is therefore an urgent need for a new canker sore medicament and patch that overcomes the above drawbacks. HBPL as a novel antibacterial and anti-inflammatory polymer material can not only effectively treat the canker sore by resisting bacteria and diminishing inflammation, but also overcome the defects of an immune accelerator or hormone medicines without bringing side effects.

Disclosure of Invention

The invention aims to solve the defects of the dental ulcer patch in the prior art and provides a slow-release hydrogel ulcer patch and a preparation method thereof. The dental ulcer patch provided by the invention has remarkable antibacterial, anti-inflammatory and analgesic effects, can be slowly released and treated for a long time without adding any hormone medicine, and has the advantages of high safety and low side effect.

The technical scheme adopted by the invention is as follows:

a slow-release type oral ulcer hydrogel ulcer patch consists of hydrogel, hyperbranched polylysine (HBPL) and other auxiliary materials, wherein the concentration of the HBPL is 10-200mg/mL, the hydrogel is hydrogel based on acrylic acid and acrylamide or derivatives thereof or is polysaccharide hydrogel, the HBPL is loaded in the hydrogel through hydrogen bond action or Schiff base action, and the HBPL causes the damage of the hydrogen bond action or the Schiff base action in the acidic environment of an oral ulcer wound, so that the HBPL is slowly released at the wound as required and is treated for a long time.

Furthermore, the hydrogel ulcer patch also contains auxiliary materials, wherein the auxiliary materials are one or more of analgesic, flavoring agent, coloring agent and antioxidant.

Further, the analgesic is one or more of ferulic acid, oleanolic acid and ursolic acid; the flavoring agent is one or more of sucrose, mannitol, sorbitol, peppermint essential oil and rose essential oil; the coloring agent is one or more of brilliant blue, lemon yellow, carmine and sunset yellow; the antioxidant is vitamins or natural plant extract, wherein the vitamins can be vitamin C or vitamin E, and the natural plant extract is tea polyphenols or grape seed extract.

Further, by adjusting the components of the hydrogel so as to adjust the strength of the hydrogen bonding or Schiff base, the release speed of the HBPL can be adjusted, and the HBPL is released more slowly when the hydrogen bonding or the Schiff base is enhanced. Thus, the ulcer patches with different components can be selected according to the severity of the dental ulcer and released as required.

According to one embodiment of the invention, a sustained-release type canker sore hydrogel ulcer patch comprises the following components: acrylamide, acrylic acid, amino-grafted hyaluronic acid (HA-ADH), aldehyde-grafted dextran (Dex-ALH), hyperbranched polylysine (HBPL), analgesics (ferulic acid, oleanolic acid, ursolic acid), flavoring agents (sucrose, mannitol, sorbitol, peppermint essential oil, rose essential oil); coloring agents (brilliant blue, lemon yellow, carmine, sunset yellow); antioxidants (vitamins can be vitamin C or vitamin E, tea polyphenols or grape seed extract);

further, the hydrogel is based on acrylic acid and acrylamide or derivatives thereof, and the preparation method comprises the following steps: acrylamide and acrylic acid are used as raw materials, ammonium Persulfate (APS)/tetramethyl diethylamine (TEMED) is used as an initiator, and dimethylformamide (MBA) is used as a cross-linking agent for synthesis; firstly, dissolving acrylamide and acrylic acid in water under magnetic stirring to obtain a solution with the weight percent of 10-30%, keeping the mass ratio of the acrylic acid to the acrylamide to be 2:10-2:20, then sequentially adding MBA, APS and TEMED, obtaining a uniform solution under magnetic stirring, and polymerizing to obtain the hydrogel.

Further, the hydrogel is polysaccharide hydrogel based on Schiff base effect, and the preparation method comprises the following steps: under the catalysis of (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride) and N-hydroxysulfosuccinimide, adipic acid hydrazine (ADH) is combined with carboxyl of hyaluronic acid to synthesize HA-ADH; oxidizing hydroxyl at ortho position of dextran with sodium periodate to prepare Dex-ALH; hA-ADH and Dex-ALH are respectively dissolved in a certain amount of water under magnetic stirring to obtain solutions with the weight percent of 1-10%, and the solutions are mixed and incubated to obtain the hydrogel.

Further, adding HBPL and auxiliary materials into the solution before gel forming of the hydrogel, spreading the film to obtain a medicinal film, cutting and sterilizing to obtain the hydrogel ulcer patch.

In the specific example, the concentration of the acrylamide and the acrylic acid is 10-30wt%, the mass ratio of the acrylic acid to the acrylamide is kept at 2:10-2:20, the concentration of HA-ADH is 1-10wt%, the concentration of Dex-ALH is 1-10wt%, the concentration of HBPL is 10-200mg/mL, the concentration of analgesic is 1-10mg/mL, the concentration of flavoring agent is 1-10mg/mL, the concentration of coloring agent is 1-10mg/mL, and the concentration of antioxidant is 1-10mg/mL.

Still further: the concentration of the acrylamide and the acrylic acid is 15-25wt%, the mass ratio of the acrylic acid to the acrylamide is kept at 2:12-2:15, the concentration of HA-ADH is 2-5wt%, the concentration of Dex-ALH is 2-5wt%, the concentration of HBPL is 20-100mg/mL, the concentration of analgesic is 2-5mg/mL, the concentration of flavoring agent is 2-5mg/mL, the concentration of coloring agent is 2-5mg/mL, and the concentration of antioxidant is 2-5mg/mL.

According to a specific example of the invention, a preparation method of the sustained-release type canker sore hydrogel ulcer patch comprises the following steps of;

dissolving acrylamide in distilled water in half amount, adding HBPL, analgesic, correctant, colorant and antioxidant, stirring, dissolving acrylic acid in distilled water in the other half amount, mixing, removing bubbles, polymerizing, spreading film, and drying to obtain medicinal film.

Dissolving HA-ALH or Dex-ALH in half amount of distilled water, adding HBPL, analgesic, correctant, colorant and antioxidant, stirring, dissolving HA-ADH or Dex-ADH in the other half amount of distilled water, mixing, removing bubbles, spreading film, and drying to obtain medicinal film.

Drying, cutting and sterilizing the obtained medicinal film to obtain the ulcer patch.

Experiments show that the acrylic acid acrylamide hydrogel and the Schiff base hydrogel provided by the invention can effectively load and slowly release HBPL. The release time of the acrylic acid acrylamide hydrogel and the Schiff base hydrogel to HBPL can last for one week. After one week, the amount of HBPL released by the acrylic acid acrylamide hydrogel reaches 70%, and the amount of HBPL released by the Schiff base hydrogel reaches 80%.

Experiments show that the HBPL provided by the invention has remarkable antibacterial, anti-inflammatory and analgesic effects. The oral ulcer patch prepared by the invention has 100 percent of antibacterial degree to staphylococcus aureus under the dosage of 20-200mg/mL of HBPL concentration.

The beneficial effects of the invention are as follows:

the dental ulcer patch prepared by the invention strengthens the combination capability and persistence between HBPL and the dental ulcer patch, inhibits the burst release of HBPL, thereby achieving the slow release effect, greatly improving the drug effect, utilizing HBPL to perform anti-inflammatory and sterilization, simultaneously having good biocompatibility, having no obvious reduction of the curative effect after being used as a treatment material for a long time, and having obvious advantages compared with hormone drugs and immune promoters. In addition, the release speed of HBPL can be realized by adjusting the content of the components according to the requirement, and the preparation can be effective for a long time without side effects.

Drawings

FIG. 1 is a release profile of HBPL over a week in example 1;

FIG. 2 shows the anti-Staphylococcus aureus effect of the ulcer patches of examples 1 and 2;

FIG. 3 is a release profile of HBPL over a week in example 3;

FIG. 4 shows the anti-Staphylococcus aureus effect of the ulcer patches of examples 3 and 4.

Detailed Description

The following examples further illustrate the technical aspects of the present invention, but are not intended to limit the present invention.

Example 1

The hydrogel is synthesized by taking acrylamide and acrylic acid as raw materials, ammonium Persulfate (APS)/tetramethyl diethylamine (TEMED) as an initiator and dimethylformamide (MBA) as a cross-linking agent. First, acrylamide and acrylic acid were dissolved in a certain amount of water under magnetic stirring to obtain a 20wt% solution. The mass ratio of acrylic acid to acrylamide was maintained at 2:13. MBA, APS and TEMED were then added sequentially and a homogeneous solution was obtained under magnetic stirring. The mass ratios of MBA, APS and TEMED to monomer were 0.1%, 0.9% and 0.6% by weight. Then adding 20mg/mL of HBPL, 5mg/mL of analgesic, 5mg/mL of flavoring agent, 5mg/mL of coloring agent and 5mg/mL of antioxidant, uniformly stirring, polymerizing to obtain hydrogel, spreading a film, drying to obtain a medicinal film, cutting, and sterilizing to obtain the hydrogel ulcer patch.

The release profile and antimicrobial effect of HBPL in this example over one week is shown in figures 1 and 2. As shown in fig. 1, slow release of HBPL is achieved due to hydrogen bonding between the hydrogel and HBPL. Meanwhile, as shown in fig. 2 (which shows different concentrations of HBPL), the antibacterial rate of staphylococcus aureus is 100%, and the ulcer patch prepared by the embodiment has excellent bactericidal performance.

Example 2

The hydrogel is synthesized by taking acrylamide and acrylic acid as raw materials, ammonium Persulfate (APS)/tetramethyl diethylamine (TEMED) as an initiator and dimethylformamide (MBA) as a cross-linking agent. First, acrylamide and acrylic acid were dissolved in a certain amount of water under magnetic stirring to obtain a 20wt% solution. The mass ratio of acrylic acid to acrylamide was maintained at 2:15. MBA, APS and TEMED were then added sequentially and a homogeneous solution was obtained under magnetic stirring. The mass ratios of MBA, APS and TEMED to monomer were 0.1%, 0.9% and 0.6% by weight. And then adding 100mg/mL of HBPL, 5mg/mL of analgesic, 5mg/mL of flavoring agent, 5mg/mL of coloring agent and 5mg/mL of antioxidant, uniformly stirring, polymerizing to obtain hydrogel, spreading a film, drying to obtain a medicinal film, cutting, and sterilizing to obtain the hydrogel ulcer patch. The increase of the mass ratio of the acrylamide can lead to the enhancement of hydrogen bonding effect, and the release of HBPL is slower, so that the acrylamide and the acrylic acid hydrogel disclosed by the invention can realize the controlled release of antibacterial components in the ulcer patch.

Example 3

Synthesis of HA-ADH: 1.0g of HA was dissolved in a buffer (pH=6) containing 0.5M NaCl and 100mM2- (N-morpholinoethanesulfonic acid (MES) to obtain a 10mg/mL solution, to which was added a 30-fold molar excess of hydrazine Adipate (ADH). The pH of the reaction mixture was maintained at 6. 3.9g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) (2.5 mmol) and 1.15g N-hydroxysuccinimide (NHS) (10 mmol) were dissolved in the reaction mixture. After the pH was adjusted to 7.0, the reaction was allowed to proceed overnight. The solution was poured into ice-ethanol to give a precipitate of HA-ADH, which was washed 3 times with water/ethanol, through a dissolution and precipitation process. Finally, after 7 days of the dialysis procedure, lyophilization was applied to obtain the final HA-ADH. Synthesis of Dex-ALH: 1.0g of dextran was dissolved in water to obtain a 10mg/mL solution, and oxidized with 2mL of sodium periodate solution at a concentration of 0.15-3.1M. After 20 hours, an equimolar amount of diethylene glycol was added and the oxidation reaction was stopped. The solution was poured into ice-ethanol to obtain a Dex-ALH precipitate, which was washed 3 times with water/ethanol, through a dissolution and precipitation process. Finally, after 7 days of the dialysis procedure, lyophilization was applied to obtain the final Dex-ALH. Preparation of the canker sore patch: respectively dissolving HA-ADH and Dex-ALH in water to obtain 5wt% solution, then adding 20mg/mL of HBPL, 5mg/mL of analgesic, 5mg/mL of flavoring agent, 5mg/mL of coloring agent and 5mg/mL of antioxidant, stirring uniformly to obtain hydrogel, spreading the hydrogel film, drying to obtain a medicinal film, cutting, and sterilizing to obtain the hydrogel ulcer patch. As shown in fig. 3, release of HBPL is achieved due to schiff base action between the hydrogel and HBPL.

The release profile and antimicrobial effect of HBPL in this example over one week is shown in figures 3 and 4. As shown in fig. 3, slow release of HBPL is achieved due to the schiff base action between the hydrogel and HBPL. Meanwhile, as shown in fig. 4 (which shows different concentrations of HBPL), the ulcer patch can achieve 100% resistance to staphylococcus aureus due to the excellent antibacterial property of HBPL.

Example 4

Synthesis of HA-ADH and Dex-ALH were performed as in example 3.

Preparation of the canker sore patch: and respectively dissolving HA-ADH and Dex-ALH in water to obtain 7 and 5wt% solutions, then adding 100mg/mL of HBPL, 5mg/mL of analgesic, 5mg/mL of flavoring agent, 5mg/mL of coloring agent and 5mg/mL of antioxidant, stirring uniformly to obtain hydrogel, spreading a film, drying to obtain a medicinal film, cutting, and sterilizing to obtain the hydrogel ulcer patch. As the content of HA-ADH is increased, the Schiff base effect is enhanced, HBPL is slowly released, and the polysaccharide hydrogel disclosed by the invention can realize the controlled release of the antibacterial component in the ulcer patch.

Claims

1. The slow-release type oral ulcer hydrogel ulcer patch is characterized by comprising hydrogel, hyperbranched polylysine, an analgesic, a flavoring agent, a coloring agent and an antioxidant; the concentration of the hyperbranched polylysine is 10-200 mg/mL; the hydrogel is hydrogel or polysaccharide hydrogel based on acrylic acid and acrylamide; the analgesic is one or more of ferulic acid, oleanolic acid and ursolic acid; the flavoring agent is one or more of sucrose, mannitol, sorbitol, peppermint essential oil and rose essential oil; the coloring agent is one or more of brilliant blue, lemon yellow, carmine and sunset yellow; the antioxidant is vitamins or natural plant extracts, wherein the vitamins are vitamin C or vitamin E, and the natural plant extracts are tea polyphenol or grape seed extracts;

the preparation method of the ulcer patch comprises the following steps:

acrylamide and acrylic acid are used as raw materials, ammonium persulfate/tetramethyl diethylamine is used as an initiator, and dimethylformamide is used as a cross-linking agent for synthesis; firstly, dissolving acrylamide and acrylic acid in water under magnetic stirring to obtain a solution with the concentration of 10-30wt%, keeping the mass ratio of the acrylic acid to the acrylamide at 2:10-2:20, sequentially adding MBA, APS and TEMED, obtaining a uniform solution under magnetic stirring, and polymerizing to obtain hydrogel; adding hyperbranched polylysine and auxiliary materials into the solution before gel forming of the hydrogel, spreading a film to obtain a medicinal film, cutting and sterilizing to obtain a hydrogel ulcer patch;

or is:

under the catalysis of (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride) and N-hydroxysulfosuccinimide, adipic acid hydrazine is combined with carboxyl of hyaluronic acid to synthesize HA-ADH; oxidizing hydroxyl at ortho position of dextran with sodium periodate to prepare Dex-ALH; respectively dissolving HA-ADH and Dex-ALH in a certain amount of water under magnetic stirring to obtain 1-10wt% solutions, mixing and incubating to obtain hydrogel; adding hyperbranched polylysine and auxiliary materials into the solution before gel forming of the hydrogel, spreading a film to obtain a medicinal film, cutting and sterilizing to obtain the hydrogel ulcer patch.