WO2011075958A1

WO2011075958A1 - Use of toll-like receptor-3 agonist in the preparation of medicament for promoting wound healing

Info

Publication number: WO2011075958A1
Application number: PCT/CN2010/070270
Authority: WO
Inventors: 苏绍波; 林青; 王俐; 林有坤; 方丹; 房佳柱; 任向荣
Original assignee: 中山大学中山眼科中心
Priority date: 2009-12-22
Filing date: 2010-01-20
Publication date: 2011-06-30
Also published as: CN101780279B; CN101780279A

Abstract

The present invention discloses the use of a toll-like receptor-3 (TLR3) agonist in the preparation of medicament for promoting wound healing. A toll-like receptor-3 (TLR3) agonist, particularly polyriboinosinic polyribocytidylic acid, can promote wound healing and be used as the active ingredient of medicament for promoting wound healing.

Description

Application of Toll-like receptor-3 agonists in the preparation of drugs for promoting wound healing

Technical field:

The invention belongs to the field of biotechnology, and particularly relates to the use of a Toll-like receptor-3 agonist in the preparation of a medicament for promoting wound healing.

Background technique:

Self-owned human beings are traumatic. Although some diseases have been controlled and eliminated with the advancement of science and technology, the causes of trauma have increased, and trauma has become one of the main causes of harm to health and even life. Moreover, whether the wound is serious or not, there are problems of tissue defect and wound healing. At the same time, the medical treatment of the wound, except for the treatment of systemic reactions and complications, can only be completed or basicized by wound healing or wound healing. carry out. Therefore, how to speed up and high-quality wound healing has become a hot issue. At present, there are many researches on healing, especially growth factors, whose strong mitogen activity can promote wound healing. However, growth factors can also make collagen secretion strong, causing excessive hyperplasia and healing quality, and may even lead to gallbladder. Lipoma and cell malignant transformation, thus limiting its clinical application. As a complex biological process, the basic process of wound healing is roughly divided into three distinct and connected stages: 1 Inflammatory phase, which manifests as inflammatory reaction, dissolves and removes necrotic tissue; 2 Period of granulation tissue formation, this period It manifests as migration and proliferation of mesenchymal cells, fibroblasts and vascular endothelial cells in the surrounding normal tissues, and finally forms granulation tissue; 3 the stage of reconstruction of new tissue, which is characterized by fibroblasts and other proliferation and maturation. Matrix deposition and transformation and reconstruction of new tissue will eventually form fibrous scar tissue. The whole process is the result of a network of interactions between a range of different types of cells, structural proteins, growth factors and protein kinases. The local inflammatory reaction of wounds is the basis of wound healing or tissue repair, which is beneficial to the repair of wounds. Although inflammatory cells are not directly involved in filling defects, they can remove necrotic tissue and foreign bodies, prevent infection, and also secrete cells. Factors regulate the wound healing process, create conditions for repairing cell activity, regulate and guide the repair process, and there is no possibility of initiation and completion of the wound healing process without localized inflammatory response.

Infectious refractory wounds, vascular wounds and recurrent episodes of skin lesions in patients with pemphigus have been a difficult problem in clinical treatment. In addition to systemic medication to control the disease, conventional topical topical quinine peptide (recombinant human epidermal growth factor external solution) is used to accelerate healing, but the effect is limited. Current foreign source Although the growth factor has achieved initial success in clinical application, growth factors also limit the secretion of collagen, cause excessive scar hyperplasia and the quality of healing, and may even cause side effects of cholesteatoma and malignant transformation. In clinical applications. In addition, the lack of expression of refractory wound receptors, mutations, and elevated protease levels have also affected the effective role of growth factors in promoting wound healing. As a complex and continuous biological reaction process, wound healing is also an important subject involved in plastic surgery. How to accelerate the healing and inhibit the formation of abnormal structures such as pathological scars is a problem that plagues the surgical community.

Polyriboinosinic polyribocytidylic acid is a synthetic double-stranded RNA polymer. Davies and Rich first synthesized successfully in 1958. As a macropolynucleotide, olyriboinosinic polyribocytidylic acid is a double-stranded multinuclear composed of Polyinosinic acid (Fig. 1A) and Polycytidylic acid (Fig. 1 B) single-stranded according to the principle of base pairing. Sour acid chain. The basic molecular formula of polyriboinosinic polyribocytidylic acid is: (C ₁₉ H230 ₁₄ N ₉ P ₂ )n, and the molecular weight is between 3-16 s. Its structural formula is shown in Figure 1C. Field and other experiments in 1967 proved that polyriboinosinic polyribocytidylic acid is a synthetic interferon strong inducing agent. In 1969, the Chinese Academy of Sciences collaborated with relevant units to develop a new polyriboinosinic polyribocytidylic acid complex preparation, which is based on polyriboinosinic polyribocytidylic acid. Kanamycin sulfate and Cacl ₂ containing polyamine-based positive ions are added to protect the double-stranded structure from being rapidly hydrolyzed by nucleases, maintaining good stability and enabling it to perform its intended function. Currently, polyriboinosinic polyribocytidylic acid has been widely used in human medicine to treat a series of viral infections. Different viruses have different sensitivities to polyriboinosinic polyribocytidylic acid, which is recognized as the most sensitive to influenza and sore viruses. Polyriboinosinic polyribocytidylic acid has good control effects on hepatitis virus, HIV, conjunctivitis virus, foot-and-mouth disease virus, vaccinia virus, encephalomyelitis virus, encephalomyocarditis virus, Aleutian virus and Coxsackie virus. Tests have shown that polyriboinosinic polyribocytidylic acid has a better effect in preventing viral infection than treatment. With the deepening of research, it is generally believed that Polyl, in addition to indirectly producing antiviral effects as a high-efficiency interferon inducer, directly antiviral, while enhancing and regulating immune function, is also the main way to exert its antiviral effect. In addition, the pharmacological effects of polyriboinosinic polyribocytidylic acid have been found to include anti-tumor, anti-protozoal and antibacterial effects.

As a ligand for type 3 Toll-like receptor-3 (TLR3) in animals, it has been reported that polyriboinosinic polyribocytidylic acid can mediate a series of exemptions after activation of TLR3. Epidemiological reactions, such as the induction of interferon, tumor necrosis factor and other cytokines and secretion of chemokines such as IL-8, MCP-1 or RANTES, promote mononuclear macrophages, NK cells, T lymphocytes, neutrophils and trees. The proliferation and maturation of a cell, etc., promotes the production of antibodies in the body. Polyriboinosinic polyribocytidylic acid has a good effect on the specific immunity and non-specific immunity of the body.

As one of the largest and most important organs of the human body, the skin is the natural barrier of the body and has important protective functions. Mikiko Tohya et al found that the expression of the epidermal chemokine MIP-Ια was up-regulated in human herpes zoster, and further experiments suggested that polyriboinosinic polyribocytidylic acid can activate the TLR3 pathway in human normal epidermal layer to promote the secretion of ΜΙΡΙα and play a role in mediating the epidermal immune response. Similarly, it was found that stratum corneum epithelial cells isolated from the lesions of patients with psoriasis can release a series of pro-inflammatory cytokines and chemokines under the stimulation of polyriboinosinic polyribocytidylic acid. Recent animal experiments have found that polyriboinosinic polyribocytidylic acid stimulates primary cultured keratinocytes to secrete pro-inflammatory factors such as TNFalpha, further demonstrating that activation of the TLR3 pathway is required for normal immune responses in damaged skin in mice.

Summary of the invention:

It is an object of the present invention to provide the use of a Toll-like receptor-3 agonist for the preparation of a medicament for promoting wound healing.

The present inventors have experimentally confirmed that a Toll-like receptor-3 agonist can promote wound healing, and thus can be used for preparing a drug for promoting wound healing.

The Toll-like receptor-3 is preferably olyriboinosinic polyribocytidylic acid, which is most effective for skin or mucosal wounds.

The wound of the present invention may be a wound caused by skin or mucosal ulcer or erosion; or a wound caused by vasculitis, diabetes, pemphigus, acne, AIDS, or wound healing; or wounds caused by burns, burns, and surgery After the wound.

The medicament may be formulated into an injection, a spray, a drop, an emulsion or a cream.

Another object of the present invention is to provide a medicament for promoting wound healing, which comprises

A Toll-like receptor-3 (TLR3) agonist is used as an active ingredient.

The polyriboinosinic polyribocytidylic acid in the present invention belongs to the prior art and can be purchased from the market.

The inventors found in the C57BL/6 mouse animal model that surgical wounds were applied in C57BL/6 mice. In the group treated with polyriboinosinic polyribocytidylic acid, the polyriboinosinic polyribocytidylic acid treatment group significantly promoted wound healing compared with the vehicle control group, and no scar was found on the skin after healing.

The inventors also conducted clinical trials: after laser spotting in patients with pigmented plaques, they were divided into two groups. One group was treated with polyriboinosinic polyribocytidylic acid injection, and the other group was treated with normal saline as a control. During the period, the patient's wound was coated with chlortetracycline ointment to promote healing; patients with vasculitis were also divided into two groups, one group of skin lesions were treated with polyriboinosinic polyribocytidylic acid injection, and the other group was treated with extravasation of gold factor peptide. (Recombinant human epidermal growth factor external solution) for treatment; Peel wounds in patients with acne are also divided into two parts, part of the lesions are treated with polyriboinosinic polyribocytidylic acid injection, and another part of the lesions are externally applied with gold peptide (recombination) Human epidermal growth factor external solution) is treated. The experimental results showed that the polyriboinosinic polyribocytidylic acid injection group significantly shortened the complete healing time of the wound compared with the conventional medicinal chlortetracycline ointment or the golden factor peptide treatment group, and the wounded skin showed no scar and the wound healing quality was high.

The Toll-like receptor-3 (TLR3) agonist of the present invention can significantly promote wound healing, especially polyriboinosinic polyribocytidylic acid can significantly promote the healing of skin or mucosal wounds, not only the healing time is significantly shortened, but also in the course of clinical treatment. No itching, redness, heat pain or other adverse reactions, and no obvious scars and skin texture changes after healing, the wound healing quality is high, thus overcoming the shortage of drugs such as clinical use of exogenous growth factors, can be used as an alternative to recombinant growth factor . Moreover, the polyriboinosinic polyribocytidylic acid of the invention has abundant sources, simple preparation process, pleasant price and convenient use. Therefore, the polyriboinosinic polyribocytidylic acid of the present invention can be used as an agent for treating wound healing, and is expected to be used for normal individuals, individuals with difficult wounds, or conventional medicine for cosmetic surgery. BRIEF DESCRIPTION OF THE DRAWINGS:

Figure 1 is a chemical structure diagram of olyriboinosinic polyribocytidylic acid;

Figure 2 is a graph showing the effect of lmg/ml polyriboinosinic polyribocytidylic acid on wound healing in mice. Figure 3 is a graph showing the effect of anti-MIP-2 antibody inhibiting lmg/ml polyriboinosinic polyribocytidylic acid on wound healing in mice.

Figure 4 is a graph showing the effect of specific TRIF-siR A on delayed wound healing in mice.

Figure 5 is a graph showing the effect of TLR3 gene deletion on wound healing in mice. Figure 6 is a graph showing the effect of lmg/ml polyriboinosinic polyribocytidylic acid on the wound effect of TLR3 knockout mice.

Figure 7 shows the effect of continuous topical lmg/ml polyriboinosinic polyribocytidylic acid injection for 5 days after facial laser pigmentation in patients with facial pigmentation.

Figure 8 shows the results of the comparison between the efficacy of the vasculitis patients and the clinical routine medication after 5 days of injection of lmg/ml polyriboinosinic polyribocytidylic acid.

Figure 9 shows the results of the comparison between the efficacy and clinical routine use of lmg/ml polyriboinosinic polyribocytidylic acid injection in patients with pemphigus in the neck and trunk.

detailed description:

The invention is further illustrated by the following examples. These embodiments are only intended to illustrate the invention, and are not intended to limit the invention, and the invention is intended to be within the scope of the invention.

I. Animal wound healing model test:

1. Formation of surgical wounds:

The animal selected first in the present invention is a healthy C57BL/6 mouse, 6-8 weeks old, half male and half female. The entire surgical process is strictly in accordance with AREB 102 and is highly vigilant. The surgery was performed in the surgical room of CAF, and the mice were intraperitoneally injected with 4% chloral hydrate to anesthetize them. The back skin surface hair was removed by 8% sodium sulfide and then sterilized with 70% alcohol and povidone iodine. Then use a sterilized, disposable skin biopsy drill to drill the entire layer of the skin on the back of the anesthetized mouse, with the midline as the central axis, and drill the skin four times on the left and right sides to prepare a skin defect with a diameter of 4 mm. Four full-thickness skins are then sterilized. The wounds are at least 1 cm apart from the intact skin. The wounds are then disinfected once daily with povidone-iodine or iodophor.

In the present invention, a T57 knockout mouse of C57BL/6 background was also used to prepare a wound animal healing model, and the selected sex of the knocking gene mouse and the wound preparation method were the same as those of the above C57BL/6 wild type mouse.

2, polyriboinosinic polyribocytidylic acid (purchased from sigma company, Product Number:

P0913) Solution configuration:

Olyriboinosinic polyribocytidylic acid (sigma) 50mg was added to a 50ml solution of endotoxin deionized water in PBS, and then the solution was dispensed into an EP tube and stored at -20 ° C for low temperature. The reagent was rewarmed to room temperature about 25 °C before use for the animal wound healing model test. 3, data analysis

The extent of wound healing was expressed as a relative percentage of the relative closed wound and the original wound (the wound just prepared on the first day, without any treatment) for each wound in each mouse. The average percentage of wound healing closure was the same for the same treatment group. The average of the percentage of wound area per day of the mouse relative to the initial wound area. In this way, the average wound healing per day for each group of mice can be graphed and compared to other treatment groups. The standard error of the average daily result can also be calculated. Example 1

Healthy C57BL/6 mice that formed surgical wounds according to the above method were randomly assigned to different treatment groups, with 6 in each group, and the researchers were unable to distinguish them during the entire observation period. After surgery, 20 μl of lmg/ml polyriboinosinic polyribocytidylic acid solution was applied topically to the left wound surface of the back skin of each experimental rat with a pipette, and the agent was spread to the entire wound with the edge of the pipette, and the right two The wound was administered the same volume of control pure PBS solution and treated as described until the wound was completely healed. On the 1st, 3rd, 5th, 7th, 12th, and 14th day after the operation, an electronic photograph of the wound was taken macroscopically, and then the wound area was quantified by using Adobe PhotoShop (version 7.0; Adobe Systems) software, each wound area. The degree of healing is expressed as a percentage relative to the initial wound. All experiments were repeated 2-3 times to ensure the stability of the experimental results, because there were differences between the different groups of mice in terms of wound healing rate, and each experiment had a solvent (PBS) control group.

Histological research:

On days 3, 5, 7, 10, 12, 14 days, after the mice in the above wound healing model were anesthetized with carbon dioxide, the wound skin tissue was cut, including the entire injured area and the muscles of the lower layer (8 mm in diameter), and then Place in 10% formalin phosphate buffer for 1 week. Histological observation: Elevated concentrations of alcohol and xylene, tissue dehydration: embedded in paraffin; cut into 5 mm thick sections placed on glass slides: stained with hematoxylin and eosin (H&E). All sections were placed under a dark field microscope and all significant changes were photographed.

The results are shown in Fig. 2. As can be seen from Fig. 2, surgical wounds of C57BL/6 mice were treated with polyriboinosinic polyribocytidylic acid daily. Our experiments confirmed that topical administration of polyriboinosinic polyribocytidylic acid once a day compared with the vehicle control group (Fig. 2, p<0.05, n=12) enhanced wound healing. Figure 2 also shows that the polyriboinosinic polyribocytidylic acid treatment group It took 10 days for the wound to heal completely, while the solvent control group (p<0.01, n=12) took 12 days, and no scar was seen on the skin after healing. Compared with the mice in the vehicle control group, the daily application of lmg/ml of polyriboinosinic polyribocytidylic acid on the wound closure from the third day after the injury, the effect is very obvious, especially on the seventh day after injury, from the wound histology Observations can also confirm this finding.

The wound skin samples of mice were collected from different days after injury, and the number of leukocytes accumulated in the wound site was counted by immunostaining. The expression of mRNA and protein levels of chemokines involved in wound healing was detected by real-time quantitative PCR and ELISA. The results showed that the number of macrophages in the wound tissue of polyriboinosinic polyribocytidylic acid was significantly increased, and among many chemokines, polyriboinosinic polyribocytidylic acid significantly promoted the synthesis of MIP-2 in wounds. Pre-administration of 100 g/ml anti-MIP-2 antibody significantly inhibited the effect of polyriboinosinic polyribocytidylic acid on wound healing, as shown in Figure 3. The results indicated that polyriboinosinic polyribocytidylic acid mediates the inflammatory response of local tissue in the wound by promoting macrophage infiltration and inducing the production of chemokine MIP-2, which affects the whole healing process.

The MyD (myeloid differentiation factor) 88-dependent signaling pathway is a common pathway for TLR signaling, while the TLR3 signaling pathway is predominantly through another MyD88-independent pathway, where TRIF [TIR(Toll-IL-1 receptor) domain -containing adapter inducing IFN-[beta] is a key adaptor protein of this TLR3-specific signaling pathway. R A interference (RNAi) is a targeted post-transcriptional gene silencing mechanism that can effectively inhibit gene expression. At present, the local delivery mode of naked siRNA [referring to the lack of specially modified or coupled siRNA (small interfering RNA) directly dissolved in a simple solution] has become a major trend in the delivery of siRNA animals due to its ease of preparation and use. Significant targeted gene silencing has been reported by intraocular injection, nasal or tracheal local administration, and targeted neurological targeting, and intramuscular, subcutaneous, tympanic, and topical mucosal epithelial administrations have also been reported. An effective in vivo delivery route for naked siRNA.

Therefore, since polyriboinosinic polyribocytidylic acid is one of the agonists of TLR3, we next use the TRIF-siRNA local wound administration method that specifically interferes with the expression of TRIF gene to preliminarily elucidate whether the activation of TLR3 signaling pathway is in the process of wound healing. It plays an important role. The TRIF-specific targeting siRNA was designed against the mouse TRIF mRNA sequence (NCBI SEQ ID NO: NM-174989.3). The specific sequence of the complementary two-stage oligonucleotide fragment is: sense strand, 5'- GCUAUGUAACA CACCGCUG TT-3 '; Antisense strand, 5 ' - CAGCGGUGUGUUACAUAGC ΤΤ -3Ό negative pair The siRNA sequence was designed by Guangzhou Ruibo Biotech Co., Ltd. The sequence was: sense strand, 5'- UUCUCCGAACGUGUCACGU TT-3'; antisense strand, 5'-ACGUGACACGUUCGGAGAA TT-3'. Both TRIF-siR A and control-siRNA were synthesized and purified by Ruibo Bio. We sterilized the double-stranded siRNA in PBS without RNase and endotoxin. Each mouse was given a total of 5 μmol of TRIF-siRNA on the back of one wound, and three other wounds on the back of the mouse. The same volume of control-siRNA, PBS and 1 mg/ml of polyriboinosinic polyribocytidylic acid were administered separately, in the same manner and time as above. The results are shown in Figure 4. Local administration of TRIF-specific siRNA significantly delayed wound healing compared with PBS-administered wounds (p<0.05, n=12), while both negative control siRNA and polyriboinosinic polyribocytidylic acid promote wound healing. The effect of healing. (Because both of them act as double-stranded RNA, they can be ligands for TLR3 and activate the TLR3 pathway). This result preliminarily indicates that activation of the TLR3 signaling pathway plays an irreplaceable role in the basic process of wound healing.

Example 2

The healthy C57BL/6 mice which formed the surgical wounds and the C57BL/6 mice which formed the surgical wounds with the TLR3 gene knockout were treated with the above methods, and the natural healing process of the wounds was observed every day after surgery.

The TLR3 knockout C57BL/6 mice that formed the surgical wounds were randomly assigned to different treatment groups, 6 in each group, and the researchers were unable to distinguish them during the entire observation period. After surgery, 20 μl of lmg/ml polyriboinosinic polyribocytidylic acid solution was applied topically to the left wound surface of the back skin of each experimental rat with a pipette, and the agent was spread to the entire wound with the edge of the pipette, and the right two The wound was administered the same volume of control pure PBS solution and treated as described until the wound was completely healed.

On the 1st, 3rd, 5th, 7th, 12th, and 14th day after the operation, an electronic photograph of the wound was taken macroscopically, and then the wound area was quantified by using Adobe PhotoShop (version 7.0; Adobe Systems) software, each wound area. The degree of healing is expressed as a percentage relative to the initial wound. All experiments need to be repeated 2-3 times to ensure the stability of the experimental results.

As shown in Fig. 5, after preparing a skin wound of the same shape and size at the same time, the photograph of the wound taken at a specific time was measured by Adobe PhotoShop software, and the wound was compared with the wild type C57BL/6 mouse as a control TLR3 knockout mouse. The healing was significantly delayed and the average complete healing time was delayed by two days. As shown in Figure 6, TLR3 knockout mice had no significant effect on topical administration of 1 mg/ml polyriboinosinic polyribocytidylic acid once daily.

The above experimental results show that polyriboinosinic polyribocytidylic acid promotes wound healing by activating the TLR3 signaling pathway in wound local tissue cells. The wound healing of TLR3 knockout mice is significantly delayed, further illustrating the activation of TLR3 signaling pathway in wound healing. The process plays an important role, and from another perspective, the efficacy of polyriboinosinic polyribocytidylic acid as an agonist of the TLR3 signaling pathway to promote wound healing. In summary, Toll-like receptor-3 (TLR3) agonists can activate TLR3 signaling pathway in wound local tissue cells to promote wound healing.

Second, clinical trials:

1, patient clinical data

According to the patient voluntary principle, 18 patients were selected, 18 of whom were skin wounds caused by 1064 nm laser laser freckle in patients with pigmented plaque, 6 were limb lesions in hospitalized patients with vasculitis, and 1 was day. Many patients with acne have erosional skin lesions on the neck and trunk. The selected patients had the same size and depth of skin lesions, and were comparable. Malnutrition, anemia, endocrine disorders, allergies, malignant tumors, radiotherapy and chemotherapy, severe liver and kidney diseases, and mental illness were excluded. Patients, pregnant or lactating women were not selected, and patients who had not completed the prescribed observation period were discontinued and were unable to judge the efficacy or incomplete data.

2, medication design:

The polyriboinosinic polyribocytidylic acid injection (purchased from Guangdong Bangmin Pharmaceutical Co., Ltd.) is used for antiviral. The main component is double-stranded Polyinosinic-Polycytidylic Acid polymer. The auxiliary material is sodium chloride, and the properties are colorless and clear liquid. Specifications: 2ml _: 2mg (concentration is 1mg/ml).

Example 3:

Patients with pigmented plaques were randomly divided into two groups according to the order of their visit. One group of patients, one day after laser debridement, was treated with lmg/ml polyriboinosinic polyribocytidylic acid injection once a day, and another group of pigmented spots was treated with postoperative wounds. Ordinary saline injection was used as a control, and the patient's wound was further coated with chlortetracycline ointment to promote healing at other times of the day.

As shown in Fig. 7, the results of clinical trials showed that compared with the clinical routine drug (chlortetracycline ointment) control group (p < 0.01, n=9), the polyporoinosinic polyribocytidylic with a partial use specification of 1 mg/ml was used. Acid injection can effectively promote wound healing. As shown in Table 1, polyriboinosinic polyribocytidylic acid significantly reduced the time to complete wound healing in patients. The average clinical control group required 7.6 days, and the olyriboinosinic polyribocytidylic acid treatment group took an average of 5.3 days, and no obvious scar residue was observed. Table 1: 10 cases of outpatients after laser ecchymosis, external use of lmg / ml polyriboinosinic polyribocytidylic acid injection, the efficacy of the results compared with clinical routine use of local lmg / ml polyriboinosinic topical topical saline injection + chlortetracycline Ointment polyribocytidylic acid injection

Patient's age preoperative diagnosis and rehabilitation patient ¥ ' Pre-existing diagnosis

(years) off (days) (years) off (days)

1 43 pigment spots 5 1 5 pigment spots 7

2 11 Ota 5 2 18 Ota 10

3 14 pigmented spots 6 3 14 tattoos 7

4 17 Ota 5 4 19 Ota 7

5 9 pigment spots 5 5 2 pigment spots 5

6 22 pigmentation spots 5 6 35 pigment spots 7

7 20 pigment spots 4 7 31 pigment spots 9

8 48 pigment spots 7 8 25 pigment spots 9

9 38 Freckles 6 9 17 Pigment spots 7

—— ¥ 1 5.3 Average 7.6 Example 4:

Inpatients with vasculitis were randomly divided into two groups. One group of patients was treated with gauze soaked with lmg/ml polyriboinosinic polyribocytidylic acid injection, and then coated with plastic film to wrap the wound. In another group of patients with vasculitis, the lesions were treated with gold-infusing peptide (recombinant human epidermal growth factor external solution) soaked gauze as control. The lesions of both groups were cleaned and changed every day.

As shown in Fig. 8, the results of the experiment showed that it was injected with olyriboinosinic polyribocytidylic acid compared with the conventional control drug (jinin peptide, recombinant human epidermal growth factor) solution (p<0.01, n=6). Fluid treatment of limb lesions in patients with vasculitis can also be very effective in promoting wound healing. As shown in Table 2, wounds treated with polyriboinosinic polyribocytidylic acid injection required an average of 21.7 days for wound healing, while the conventional control group required an average of 26.25 days. Table 2: 4 patients admitted to hospital for vasculitis, limb wound lesions external use lmg / ml polyriboinosinic polyribocytidylic acid injection, its efficacy compared with clinical routine medication results with lmg / ml polyriboinosinic

Polyribocytidylic acid injection (recombinant human epidermal growth factor external solution) patient age (years) healing (days) patient age (years) healing (days)

1 41 24 1 55 29

2 56 26 2 51 30

3 51 20 3 44 27

4 58 17 4 49 19

Average 21.75 Average 26.25 Example 5:

There was a case of pemphigus, a history of pemphigus for 2 years, long-term treatment with glucocorticoids, neck erosion has not healed for three months. Two months before the treatment with the drug of the present invention, admission to the hospital for upper gastrointestinal bleeding, the admission examination found that the biochemical examination was consistent with the diagnosis of diabetes, confirmed that he had a history of diabetes, had stopped using hormone for 8 days, and the neck erosion was aggravated. Blisters also appear on the back, smashed. After daily dexamethasone 5mg intravenous use for 2 months, no new rash appeared, the wound was washed daily, but the erosion did not heal, plus the use of Jinin peptide (recombinant human epidermal growth factor external solution) for 2 weeks, the erosion improved, but Still unhealed. In the neck and torso wounds, the gold peptide was discontinued. Polyriboinosinic polyribocytidylic acid injection was applied to the wound, and the gold peptide was continued at the low tail. The comparison photos are collected as shown in Figure 9. In the treatment area of polyriboinosinic polyribocytidylic acid injection, skin erosion has been basically cured, and lesions are still visible in the treatment of the gold peptide.

In summary, the present inventors have observed that in the animal model and clinical trials, topical application of polyriboinosinic polyribocytidylic acid can significantly accelerate the normal wounds of model mice, as well as recurrent wounds such as vasculitis or pemphigus and laser ecchymosis. Wounds such as wounds heal without accompanying excessive hyperplasia of the scar. Therefore, polyriboinosinic polyribocytidylic acid is expected to be a routine drug for normal individuals, individuals with unhealthy wounds, or individuals undergoing cosmetic surgery.

Claims

Rights request

1. Use of a Toll-like receptor-3 agonist for the preparation of a medicament for promoting wound healing.

2. The use according to claim 1, wherein the Toll-like receptor-3 agonist is Polyriboinosinic polyribocytidylic acid.

3. Use according to claim 1 or 2, characterized in that the wound is a skin or mucous membrane wound.

4. The use according to claim 3, wherein: the wound is a wound caused by ulceration or erosion of the skin or mucous membrane; or the wound does not heal by vasculitis, diabetes, pemphigus, acne, AIDS And the resulting wound; or wounds caused by burns, burns, and wounds after surgery.

5. Use according to claim 1 or 2, characterized in that the pharmaceutical dosage form is an injection, a spray, a drop, an emulsion or a cream.

A drug for promoting wound healing, characterized in that the drug contains a Toll-like receptor-3 agonist as an active ingredient.

The medicament according to claim 6, wherein the Toll-like receptor-3 agonist is Polyriboinosinic polyribocytidylic acid.