Application of micromolecular hyaluronic acid in preparation of medicine for preventing or treating virus infection with hemagglutinin on surface

Abstract

The invention discloses application of micromolecular hyaluronic acid in preparation of a medicine for preventing or treating virus infection with hemagglutinin on the surface. At least part of the small molecular hyaluronic acid inhibits the hemagglutinin of the virus from being combined with the cell surface through a mechanism of combining or interacting with the erythrocyte surface so as to prevent or treat virus infection; the small molecular hyaluronic acid is small molecular hyaluronic acid with the average molecular weight of 35 kDa; or the small molecular hyaluronic acid is small molecular hyaluronic acid with the average molecular weight of 70kDa; or the small molecular hyaluronic acid is small molecular hyaluronic acid with the average molecular weight of 24kDa. The invention also discloses a biological activity detection method of the micromolecular hyaluronic acid for preventing or treating virus infection with hemagglutinin on the surface, and biological activity detection is carried out by adopting an in-vitro test of inducing erythrocyte aggregation by the micromolecular hyaluronic acid. The invention provides a new application direction for small molecular hyaluronic acid; or a novel prevention or treatment medicine is provided for virus infection with hemagglutinin on the surface.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to the use of small molecule hyaluronic acid in the preparation of a drug for preventing or treating viral infection with hemagglutinin on the surface. Background Art

[0002] Viral infections, especially influenza virus infections, are very harmful to the human body. Taking influenza virus as an example, autumn and winter are the peak seasons for influenza. Compared with the common cold, influenza is characterized by its strong infectiousness and more severe symptoms, which can cause more serious systemic symptoms, such as fatigue, body aches, and even death. The main pathogen causing influenza is influenza virus. There is a glycoprotein on the surface of influenza virus, namely hemagglutinin (HA). There are three main subtypes of influenza hemagglutinin known to humans, labeled H1, H2 and H3.

[0003] Hemagglutinin exists on the surface of influenza viruses and plays an important role in the influenza virus infection process. Influenza viruses use hemagglutinin as a medium to adsorb on the cell surface, which is the first step for influenza viruses to infect the human body. For example, influenza viruses adhere to the surface of respiratory epithelium through the hemagglutinin on their envelope, inducing epithelial cells to engulf the virus through endocytosis and replicate inside the cells. In other words, the hemagglutinin on the surface of influenza viruses is erythrocyte hemagglutinin, which can induce erythrocyte agglutination, and through this mechanism, infect and enter upper respiratory tract cells, forming the pathogenic effect of influenza viruses. The pathogenic mechanisms of other viruses with hemagglutinin on their surfaces are similar. At present, there are basically no non-vaccine and antibody treatment options specifically for viruses, especially influenza viruses, that specifically inhibit the binding of hemagglutinin to the host cell surface.

[0004] The main therapeutic effects of small molecule hyaluronic acid include anti-inflammatory, analgesic, antipruritic, beauty and anti-aging. So far, there have been no reports at home and abroad on the use of small molecule hyaluronic acid to prevent viruses, especially diseases caused by influenza virus infection. Summary of the invention

[0005] The first technical problem to be solved by the present invention is to provide an application of a small molecule hyaluronic acid in the preparation of a drug for preventing or treating viral infection with hemagglutinin on the surface, providing a new application direction for small molecule hyaluronic acid; or to provide a new preventive or therapeutic drug for viral infection with hemagglutinin on the surface.

[0006] The second technical problem to be solved by the present invention is to provide a method for detecting the biological activity of small molecule hyaluronic acid for preventing or treating viral infection with hemagglutinin on the surface, so that the biological activity can be detected conveniently and quickly in vitro.

[0007] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a use of a small molecule hyaluronic acid in the preparation of a drug for preventing or treating viral infection with hemagglutinin on its surface.

[0009] As a further improvement of the present invention, the small molecule hyaluronic acid at least partially inhibits the binding or interaction of viral hemagglutinin with the surface of red blood cells, thereby preventing or treating viral infection.

[0010] Furthermore, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 35 kDa, wherein >90% of the molecular weight is distributed in the range of 10-70 kDa;

[0011] Or, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 70 kDa, wherein >90% of the molecular weight is distributed in the range of 40-100 kDa;

[0012] Alternatively, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 24 kDa, wherein >90% of the molecular weight is distributed between 5-45 kDa.

[0013] Furthermore, the virus having hemagglutinin on its surface is an influenza virus having hemagglutinin on its surface.

[0014] Furthermore, the virus having hemagglutinin on its surface is hepatitis B virus, rabies virus, forest encephalitis virus or Japanese encephalitis virus having hemagglutinin on its surface.

[0015] Furthermore, the medicine is an injection or an oral preparation.

[0016] In a second aspect, the present invention provides a method for detecting the biological activity of small molecule hyaluronic acid for preventing or treating viral infection with hemagglutinin on the surface, and the biological activity is detected by an in vitro test of small molecule hyaluronic acid inducing red blood cell aggregation.

[0017] Furthermore, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 35 kDa, wherein >90% of the molecular weight is distributed in the range of 10-70 kDa;

[0018] Or, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 70 kDa, wherein >90% of the molecular weight is distributed in the range of 40-100 kDa;

[0019] Alternatively, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 24 kDa, wherein >90% of the molecular weight is distributed between 5-45 kDa.

[0020] The present invention verifies through experiments that small molecule hyaluronic acid, like the hemagglutinin of viruses (especially influenza viruses), induces agglutination of erythrocytes of humans and various animals by binding to or interacting with the surface of erythrocytes. The present invention further finds that small molecule hyaluronic acid inhibits the binding or interaction of viral hemagglutinin with the surface of erythrocytes at least in part by binding to or interacting with the surface of erythrocytes, thereby preventing or treating viral infection, that is, small molecule hyaluronic acid inhibits the binding of viral hemagglutinin with the cell surface at least in part by binding to or interacting with the surface of erythrocytes, thereby preventing or treating viral infection, and small molecule hyaluronic acid is a drug that prevents and treats viral infection by blocking the adsorption or interaction of viruses on the cell surface. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The above is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention, the present invention is further described in detail below in conjunction with the accompanying drawings and specific implementation methods.

[0022] Figure 1 This is a graph evaluating the weight change or morbidity of influenza mice after intraperitoneal injection of BALB / C mice with 35kDa and 70kDa small molecule hyaluronic acid;

[0023] Figure 2 This is a graph evaluating the survival rate of influenza mice after intraperitoneal injection of BALB / C mice with small molecule hyaluronic acid with an average molecular weight of 35kDa and 70kDa. DETAILED DESCRIPTION

[0024] The present invention has found through research that small molecule hyaluronic acid inhibits the binding or interaction of viral hemagglutinin with the surface of red blood cells at least in part by binding or interacting with the surface of red blood cells, thereby preventing or treating viral infection.

[0025] Among them, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 35kDa, wherein >90% of the molecular weight is distributed between 10-70kDa; or, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 70kDa, wherein >90% of the molecular weight is distributed between 40-100kDa; or, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 24kDa, wherein >90% of the molecular weight is distributed between 5-45kDa. The above small molecule hyaluronic acids are all known products, generally in powder form, and can be made into injections or oral preparations.

[0026] Among them, the virus with hemagglutinin on the surface is influenza virus with hemagglutinin on the surface; or the virus with hemagglutinin on the surface is hepatitis B virus, rabies virus, forest encephalitis virus or Japanese encephalitis virus, etc. with hemagglutinin on the surface.

[0027] Based on the above research results, the present invention also provides a method for detecting the biological activity of small molecule hyaluronic acid for preventing or treating viral infection with hemagglutinin on the surface, which mainly uses an in vitro test of small molecule hyaluronic acid inducing red blood cell aggregation to perform biological activity detection.

[0028] Since the main purpose of the present invention is to find that small molecule hyaluronic acid inhibits the binding of viral hemagglutinin to the red blood cell surface by binding to the red blood cell surface, thereby preventing or treating viral infection, therefore, any viral infection with hemagglutinin can be treated with the above method.

[0029] The following is mainly based on the influenza virus as an example for detailed description, but it should not be limited to this virus. The present invention first studies the preventive and therapeutic effects of small molecule hyaluronic acid with an average molecular weight of 35kDa on influenza in mice. Then, the present invention retrospectively studies the situation of 3 volunteers of different ages and genders who have used small molecule hyaluronic acid with an average molecular weight of 35kDa for a long time and have not used influenza vaccines for more than 9 years. At the same time, this study also introduces 2 cases of effective oral treatment of influenza using small molecule hyaluronic acid with an average molecular weight of 70kDa and 35kDa.

[0030] Example 1

[0031] Objective: The effect of hyaluronic acid with average molecular weight of 24kDa, 35kDa and 70kDa on inducing erythrocyte aggregation in humans and animals is mutually inhibited by the effect of hemagglutinin on the influenza virus envelope on inducing erythrocyte aggregation in humans and animals.

[0032] background:

[0033] Hemagglutinin on the influenza virus envelope is also called hemagglutinin, which induces red blood cell agglutination. Hemagglutinin on the surface of influenza virus is a sugar-containing protein that can induce red blood cell aggregation in humans and animals (https: / / en.wikipedia.org / wiki / Hemagglutinin). This phenomenon of inducing red blood cell aggregation in humans and animals is used to study the activity of influenza virus. The influenza virus adheres to the surface through the hemagglutinin on its envelope, inducing the influenza virus to enter the respiratory epithelial cells. In other words, the influenza virus infects respiratory cells through this mechanism, forming the pathogenic effect of influenza virus. This example studies whether the effect of hyaluronic acid with an average molecular weight of 24kDa, 35kDa and 70kDa on inducing red blood cell aggregation in humans and animals and the effect of hemagglutinin on the influenza virus envelope on inducing red blood cell aggregation in humans and animals have a mutually inhibitory relationship. In other words, this example assumes that the average molecular weight of 24kDa, 35kDa and 70kDa small molecule hyaluronic acid and the hemagglutinin on the influenza virus envelope are bound to the surface of red blood cells and respiratory cells.

[0034] method:

[0035] Erythrocyte aggregation study: The average molecular weight of 24kDa, 35kDa and 70kDa small molecule hyaluronic acid was mixed with the venous blood of humans, mice, beagles, sheep, cattle, pigs, alpacas, horses, minks, monkeys and cats to a final concentration of 12mg / mL, 6mg / mL, 3mg / mL, 1.5mg / mL, 0.75mg / mL and 0.375mg / mL, respectively. Erythrocyte smears were prepared and the roule-like aggregation of erythrocytes induced by the average molecular weight of 24kDa, 35kDa and 70kDa small molecule hyaluronic acid was observed under a microscope. The lowest concentration that induced the aggregation of erythrocytes in humans, mice, beagles, sheep, cattle, pigs, alpacas, horses, minks, monkeys and cats was obtained. The results are shown in Tables 1, 2 and 3.

[0036] result:

[0037] The present invention found that 24kDa, 35kDa and 70kDa small molecular weight hyaluronic acid induces erythrocyte aggregation like influenza hemagglutinin. The following are the research results on the average molecular weight 24kDa, 35kDa and 70kDa inducing erythrocyte aggregation of different animals (Tables 1, 2, 3).

[0038] Table 1. Minimum concentration of 24kDa small molecule hyaluronic acid fragments that induce red blood cell aggregation.

[0039]

[0040]

[0041] Note: Low concentration of 24kDa small molecule hyaluronic acid did not induce red blood cell aggregation in cattle and sheep, suggesting that 24kDa small molecule hyaluronic acid is species-specific in preventing animal influenza.

[0042] Table 2. Minimum concentration of 35kDa small molecule hyaluronic acid fragments that induce red blood cell aggregation.

[0043]

[0044] Note: Low concentration of 35kDa small molecule hyaluronic acid did not induce red blood cell aggregation in cattle and sheep, suggesting that 35kD small molecule hyaluronic acid is species-specific in preventing animal influenza.

[0045] Table 3. Minimum concentration of 70kDa small molecule hyaluronic acid fragments that induce red blood cell aggregation.

[0046]

[0047] Note: Low concentration of 70kDa small molecule hyaluronic acid did not induce red blood cell aggregation in cattle and sheep, suggesting that 70kDa small molecule hyaluronic acid is species-specific in preventing animal influenza.

[0048] discuss:

[0049] The phenomenon of red blood cell aggregation is usually caused by the hemagglutinin (a substance that promotes blood coagulation) on the surface of the virus binding to the surface of red blood cells, which leads to enhanced interaction between red blood cells and ultimately forms red blood cell agglutination or precipitation. In addition, the study of virus-induced red blood cell aggregation can also help to deepen the understanding of the interaction mechanism between viruses and host cells, and provide a theoretical basis for the development of antiviral drugs and vaccines.

[0050] Influenza virus adheres to the surface of host cells through the hemagglutinin on its envelope, inducing influenza virus to enter respiratory epithelial cells. In other words, influenza virus infects respiratory cells through this mechanism, forming the pathogenic effect of influenza virus. The hemagglutinin on the influenza virus envelope is also called hemagglutinin, which induces hemagglutination of red blood cells. The present invention found that the average molecular weight of 24kDa, 35kDa and 70kDa small molecule hyaluronic acid induces hemagglutination of red blood cells like influenza hemagglutinin (Table 1, 2, 3). The present invention hypothesizes that the average molecular weight of 24kDa, 35kDa and 70kDa small molecule hyaluronic acid and the hemagglutinin on the influenza virus envelope have a mutually inhibitory effect and are drugs for preventing influenza.

[0051] The results in Tables 1, 2, and 3 show that low-concentration 24kDa, 35kDa, and 70kDa small-molecule hyaluronic acid induces agglutination of human and animal erythrocytes, supporting the hypothesis that high-dose 24kDa, 35kDa, and 70kDa small-molecule hyaluronic acid can prevent influenza by simulating the agglutination of erythrocytes induced by influenza hemagglutinin. In other words, both 24kDa, 35kDa, and 70kDa small-molecule hyaluronic acid and the hemagglutinin on the influenza virus envelope bind to the cell surface.

[0052] Influenza virus adsorbs on the surface of host cells through the hemagglutinin on its envelope, inducing epithelial cells to engulf the virus through endocytosis and replicate inside the cell. Small molecule hyaluronic acid with average molecular weight of 24kDa, 35kDa and 70kDa replaces the hemagglutinin on the surface of influenza, blocking the first step of influenza virus infection in the human body, and becomes a flu prevention drug, including all types of influenza. Influenza virus, hepatitis B virus, rabies virus, forest encephalitis virus and Japanese encephalitis virus all have the ability to agglutinate red blood cells.

[0053] Conclusion: Small molecule hyaluronic acid with average molecular weight of 24kDa, 35kDa and 70kDa blocks the binding of influenza hemagglutinin to the cell surface and prevents the occurrence of influenza. The phenomenon of erythrocyte aggregation induced by small molecule hyaluronic acid with average molecular weight of 24kDa, 35kDa and 70kDa is a biological activity experiment of small molecule hyaluronic acid in preventing and treating influenza.

[0054] Example 2

[0055] Objective: To investigate the preventive effect of 35kDa and 70kDa small molecule hyaluronic acid on influenza in mice. Methods:

[0056] This example studies the preventive effect of average molecular weight 35kDa (>90% of the distribution range is between 10-70kDa) and average molecular weight 70kDa (>90% of the distribution range is between 40-100kDa) on influenza virus in the mouse influenza H1N1 model. 16 8-week-old female BALB / C mice weighing 18-22 grams were divided into experimental groups 1, 2 and a control group, with 8 mice in each group. The control group was injected intraperitoneally with 1.0 ml of MEM culture medium. Experimental group 1 used 2% (25mg / ml) of average molecular weight 35kDa small molecule hyaluronic acid dissolved in MEM culture medium, and each was injected with about 1250mg / kg body weight. Experimental group 2 used 2% (25mg / ml) of average molecular weight 70kDa small molecule hyaluronic acid dissolved in MEM culture medium, and each was injected with about 1250mg / kg body weight. Mice were anesthetized with ethylfluorocarbon gas 2 hours after injection. Then, 50ul of mouse-adapted influenza virus (A / PuertoRico / 8 / 34 (H1N1), PR8) was inoculated intranasally, with each mouse inoculated with 10xMLD50 influenza virus. After the challenge, the weight loss or morbidity of the mice was monitored every day. Figure 1 ) and survival rate ( Figure 2 ) was used to determine the preventive effect of small molecule hyaluronic acid with average molecular weight of 35kDa and 70kDa on influenza virus infection.

[0057] result:

[0058] The present invention hypothesizes that the small molecule hyaluronic acid with an average molecular weight of 35kDa and 70kDa produces competitive inhibition with the hemagglutinin on the influenza virus envelope, and is a candidate drug for preventing influenza. Note 1: The binding or interaction between the small molecule hyaluronic acid with an average molecular weight of 24KD and the surface of red blood cells is 3-4 times weaker than that of the small molecule hyaluronic acid with an average molecular weight of 35kDa (see Example 1), and the required dose is 3-4 times larger; Note 2: The small molecule hyaluronic acid with an average molecular weight of 35kDa described in this patent application has a molecular weight distribution of >90% between 10-70kDa, which has covered most of the small molecule hyaluronic acid with an average molecular weight of 24KD. Therefore, the present invention first studied the preventive effect of small molecule hyaluronic acid with an average molecular weight of 35kDa and 70kDa on influenza in BALB / C mice. The results of the study showed that intraperitoneal injection of small molecule hyaluronic acid with an average molecular weight of 35kDa and 70kDa is effective in preventing and treating H1N1 influenza in mice ( Figure 1 , 2 ).

[0059] The present invention studies the preventive effect of small molecule hyaluronic acid with average molecular weight of 35kDa and 70kDa on influenza H1N1 in BALB / C mice by simulating the red blood cell aggregation induced by influenza hemagglutinin. Figure 1 , 2 The incidence of influenza H1N1 in mice treated with and without 25mg / 1ml injection of 35kDa and 70kDa small molecule hyaluronic acid ( Figure 1 ) and survival rate ( Figure 2 ) evaluation. The results showed that all influenza BALB / C mice that were not treated with 35kDa and 70kDa small molecule hyaluronic acid died ( Figure 2 ), all influenza BALB / C mice treated with 25mg / 1ml injection of 35kDa and 70kDa small molecular weight hyaluronic acid did not die within 14 days of observation ( Figure 2 The results also showed that all influenza BALB / C mice treated with 25mg / 1ml injection of small molecule hyaluronic acid without death had a significant decrease in body weight ( Figure 1 ), all influenza BALB / C mice treated with 25mg / 1ml injection of 35kDa and 70kDa small molecular weight hyaluronic acid showed no significant weight loss during the 14 days of observation ( Figure 1 ). The above research results of influenza H1N1 in mice indicate that intraperitoneal injection of small molecule hyaluronic acid with average molecular weight of 35kDa and 70kDa can prevent and treat influenza H1N1 in mice.

[0060] discuss:

[0061] This example first studies the weight loss (morbidity) and survival rate of female 8-week-old BALB / C mice weighing 18-22 grams of H1N1 influenza after intraperitoneal injection of 25 mg / 1 ml of 35 kDa and 70 kDa small molecule hyaluronic acid. The 35 kDa and 70 kDa small molecule hyaluronic acid has a preventive and therapeutic effect on influenza in mice by simulating the red blood cell aggregation induced by influenza hemagglutinin. Figure 1 , 2 The results of the study on H1N1 influenza in BALB / C mice showed that intraperitoneal injection of 35kDa and 70kDa small molecule hyaluronic acid effectively prevented and treated H1N1 influenza infection in BALB / C mice ( Figure 1 , 2 ).

[0062] Conclusion: The results of this study support the prevention and treatment of influenza by injection of large doses of small molecule hyaluronic acid with an average molecular weight of 35 kDa and 70 kDa.

[0063] Example 3

[0064] Objective: To prevent volunteers from contracting influenza who frequently and long-term used low-molecular-weight hyaluronic acid (LMHA) with an average molecular weight of 35 kDa (>90% of the distribution range is between 10-70 kDa).

[0065] method:

[0066] This example retrospectively studies the influenza situation of three volunteers who have been injected with 35kD small molecule hyaluronic acid for a long time and frequently (100 mg / 5 ml, once a week) for 9 years (Tables 4 and 5). These three volunteers have lived in Qingdao and Hangzhou, China for 9 years and have never used influenza vaccines. Among them, Li, female, 71 years old, has been injected with 35kD small molecule hyaluronic acid for a long time and frequently (100 mg / 5 ml, once a week) for neck pain, eye diseases and meningioma; Wang, female, 41 years old, has been injected with 35kD small molecule hyaluronic acid for face slimming, skin beauty and shoulder pain; Hui, male, 65 years old, has been injected with 35kD small molecule hyaluronic acid for a long time and frequently (100 mg / 5 ml, once a week) for face slimming, skin beauty and shoulder pain. 100 mg / 5 ml, once a week) injection to supplement the average molecular weight of 35kD small molecule hyaluronic acid, mainly used for face slimming, skin beauty, treatment of back pain and anti-aging. These three volunteers injected 100 mg / 5 ml of 35kDa small molecule hyaluronic acid into the deep abdominal fat on average every week from the autumn and winter flu season of 2016 to 2025. The average molecular weight of 35kD small molecule hyaluronic acid is made by cutting the large molecule hyaluronic acid injection solution with recombinant human hyaluronidase PH20. Note: These three volunteers began to use a large dose of small molecule hyaluronic acid with an average molecular weight of 70kDa (>90% of the distribution range is between 40-100kDa) in 2024, 5 grams orally each time, once every 3 days.

[0067] Table 4. Major differences between influenza and the common cold.

[0068]

[0069] result:

[0070] This example then retrospectively studied the influenza incidence of three volunteers of different ages and genders who had been injected with small molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) for a long time and frequently (100 mg / 5ml, once a week) for 9 years (Table 5). The results of the study showed that these three volunteers had not had influenza in 9 years (Table 5) and had not used influenza vaccines. The results of this study support that small molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) has the effect of preventing influenza. All three volunteers had suffered from new coronary infection, and the results of nucleic acid tests were positive and accompanied by fever, sore throat and muscle aches, suggesting that small molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) has no preventive effect on new coronary pneumonia without hemagglutinin. These three volunteers started using large doses of small molecule hyaluronic acid with an average molecular weight of 70kDa (>90% of the distribution range is between 40-100kDa) at the same time in 2024, 5 grams orally each time, once every 3 days. This also suggests that large doses of 5 grams of oral supplementation of small molecule hyaluronic acid with an average molecular weight of 70kDa (>90% of the distribution range is between 40-100kDa) can also prevent influenza.

[0071] Table 5. The main types of influenza that occurred in China reported in the "China Influenza Surveillance Weekly Report" from 2016 to 2024, and the cases of influenza and COVID-19 in 3 volunteers of different ages and genders who had never used influenza vaccines and received supplementary treatment with small molecule hyaluronic acid with an average molecular weight of 70kDa.

[0072]

[0073] Note: Mild COVID-19 infections mainly rely on positive COVID-19 nucleic acid tests and are accompanied by fever, sore throat and muscle aches.

[0074] discuss:

[0075] This example retrospectively studies the incidence of influenza and new crown infection in three volunteers of different ages and genders who have been injected with small molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) for a long time and frequently (100 mg / 5ml, once a week) for 9 years (Table 4). The results of the retrospective study in this example (Table 5) show that these three volunteers have not had influenza or used influenza vaccines in 9 years. This retrospective study shows that long-term use (100 mg / 5ml, once a week) of small molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) prevents different types of influenza (Table 5) that occurred in 9 years, and is effective for different genders and ages. The results of this study support that small molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) has the effect of preventing influenza. These three volunteers had all been infected with the new coronavirus within 9 years (Table 5), indicating that small-molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) has no preventive effect on infection with the new coronavirus without hemagglutinin.

[0076] Conclusion: Injection supplementation of small molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) can prevent and treat influenza infection, suggesting that injection of small molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) and oral administration of large doses of small molecule hyaluronic acid with an average molecular weight of 70kDa (>90% of the distribution range is between 40-100kDa) are drugs for preventing and treating influenza virus infection with hemagglutinin on the surface.

[0077] Example 4

[0078] Objective: To treat severe influenza with large doses of oral small molecule hyaluronic acid with an average molecular weight of 70 kDa (>90% of the distribution range is between 40-100 kDa).

[0079] method:

[0080] This example studies a male volunteer aged 22 who suffered from severe influenza at an altitude of 5,000 meters (Tables 4 and 6). This case was treated with a large dose of oral small molecule hyaluronic acid with an average molecular weight of 70 kDa (>90% of the distribution range is between 40-100 kDa). The severity of headache, muscle aches and fatigue symptoms of severe influenza before and after treatment was estimated by the patient's self-rating of 0-10 on the Numeric Pain Rating Scale (NPRS) before and after treatment.

[0081] result:

[0082] The results of the study showed that large doses (10 g) of oral administration of small molecule hyaluronic acid with an average molecular weight of 70 kDa (>90% of the distribution range is between 40-100 kDa) are effective in treating severe influenza (Table 6).

[0083] Table 6. Improvement of influenza signs and symptoms before and after treatment with high-dose oral small molecule hyaluronic acid with an average molecular weight of 70 kDa (>90% of the distribution range is between 40-100 kDa).

[0084]

[0085] discuss:

[0086] The results of the present invention show that large doses (10 grams) of oral small molecule hyaluronic acid with an average molecular weight of 70 kDa (>90% of the distribution range is between 40-100 kDa) are effective in treating severe influenza (Table 6). Large doses of oral small molecule hyaluronic acid with an average molecular weight of 70 kDa (>90% of the distribution range is between 40-100 kDa) are promising drugs for preventing and treating influenza virus infections with hemagglutinins on the surface. Conclusion: Small molecule hyaluronic acid with an average molecular weight of 70 kDa (>90% of the distribution range is between 40-100 kDa) prevents and treats influenza virus infections with hemagglutinins on the surface.

[0087] Example 5

[0088] Objective: To treat influenza by oral administration of high doses of small molecule hyaluronic acid with an average molecular weight of 35 kDa (>90% of the distribution range is between 10-70 kDa).

[0089] method:

[0090] This example studies a case of influenza in a 42-year-old woman (Table 7). In this case, a large dose of oral small molecule hyaluronic acid with an average molecular weight of 35 kDa (>90% of the distribution range is between 10-70 kDa) was used to treat the headache, muscle aches and fatigue symptoms of influenza before and after treatment. The severity was estimated by the Numeric Pain Rating Scale (NPRS) patient self-rating of 0-10. The references are as follows: 1. Dworkin R, Turk D, Farrar JT. Core outcome measures for chronic pain clinical trials: IMMPACT recommendations. Pain. 2005; 113: 9-19.27; 2. Cleland J, Childs J, Whitman J. Psychometric properties of the neck disability index and numeric pain rating scale in patients with mechanical neck pain. Arch Phys Med Rehabil. 2008; 89: 69-74.

[0091] result:

[0092] The results of this example show that large doses (10 grams) of oral administration of small molecule hyaluronic acid with an average molecular weight of 35 kDa (>90% of the distribution range is between 10-70 kDa) are effective in treating influenza (Table 7).

[0093] Table 7. Improvement of influenza signs and symptoms before and after treatment with large doses of oral small molecule hyaluronic acid with an average molecular weight of 35 kDa (>90% of the distribution range is between 10-70 kDa).

[0094]

[0095] discuss:

[0096] The results of this example show that large doses of oral small molecule hyaluronic acid with an average molecular weight of 35 kDa (>90% of the distribution range is between 10-70 kDa) are effective in treating influenza (Table 7).

[0097] Conclusion: High-dose oral administration of small molecule hyaluronic acid with an average molecular weight of 35 kDa (>90% of the distribution range is between 10-70 kDa) can prevent and treat influenza virus infection with hemagglutinin on its surface.

[0098] Example 6

[0099] Objective: The mechanism of action of small molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) is to kill influenza viruses and prevent influenza virus infection by promoting human immunity.

[0100] Methods: Related literature research.

[0101] Results: Professor David Jackson of Oxford University was the first scientist to discover the hyaluronic acid receptor LYVE-1 in the lymphatic system and its role in promoting lymphocyte return. This work subsequently prompted Dr. Matthew Hui from the University of Toronto to discover that small-molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) is less than 220 nanometers in size and can pass through a 220-nanometer filter membrane after injection and oral administration, and can quickly return to the blood through the lymph nodes and spleen. He also found that 35kD small-molecule hyaluronic acid also promotes the movement of lymphocytes. These research results suggest that small-molecule hyaluronic acid with an average molecular weight of 35kD (>90% of the distribution range is between 10-70kDa) promotes human immunity. Professors Vera Gorbunova and Andrei Seluanov of the University of Rochester have been studying the tissue hyaluronic acid levels of naked mole rats for 15 years, and the hyaluronic acid levels of these animals can be as high as 6%. Naked mole rats have an average lifespan of 32 years, so they are called "immortal creatures." Therefore, the longevity of naked mole rats with high hyaluronic acid may also be related to their good immunity.

[0102] Conclusion: Low-molecular-weight hyaluronic acid with an average molecular weight of 35 kDa (>90% of the distribution range is between 10-70 kDa) has the effect of promoting human immunity. Combined with the above research results of the present invention, it can be seen that it may kill influenza viruses and prevent influenza virus infection.

[0103] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Those skilled in the art may make some simple modifications, equivalent changes or modifications using the technical contents disclosed above, which all fall within the protection scope of the present invention.

Claims

1. Use of a small molecule hyaluronic acid in the preparation of a drug for preventing or treating viral infection with hemagglutinin on its surface.

2. The use of the small molecule hyaluronic acid according to claim 1 in the preparation of a drug for preventing or treating viral infection with hemagglutinin on the surface, characterized in that: The small molecule hyaluronic acid at least partially inhibits the binding or interaction of viral hemagglutinin with the surface of red blood cells, thereby preventing or treating viral infection.

3. Use of the small molecule hyaluronic acid according to claim 1 in the preparation of a drug for preventing or treating viral infection with hemagglutinin on the surface, characterized in that: The small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 35 kDa, wherein >90% of the molecular weight is distributed in the range of 10-70 kDa; Or, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 70 kDa, wherein >90% of the molecular weight is distributed in the range of 40-100 kDa; Alternatively, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 24 kDa, wherein >90% of the molecular weight is distributed between 5-45 kDa.

4. Use of the small molecule hyaluronic acid according to any one of claims 1 to 3 in the preparation of a drug for preventing or treating viral infection with hemagglutinin on the surface, characterized in that: The virus with hemagglutinin on the surface is an influenza virus with hemagglutinin on the surface.

5. Use of the small molecule hyaluronic acid according to any one of claims 1 to 3 in the preparation of a drug for preventing or treating viral infection with hemagglutinin on the surface, characterized in that: The virus with hemagglutinin on the surface is hepatitis B virus, rabies virus, forest encephalitis virus or Japanese encephalitis virus with hemagglutinin on the surface.

6. Use of the small molecule hyaluronic acid according to any one of claims 1 to 3 in the preparation of a drug for preventing or treating viral infection with hemagglutinin on the surface, characterized in that: The medicine is an injection or an oral preparation.

7. A method for detecting the biological activity of small molecule hyaluronic acid for preventing or treating viral infection with hemagglutinin on its surface, characterized in that: The biological activity was detected by using an in vitro test of small molecule hyaluronic acid inducing red blood cell aggregation.

8. The method for detecting the biological activity of small molecule hyaluronic acid for preventing or treating viral infection with hemagglutinin on the surface according to claim 7, characterized in that: The small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 35 kDa, wherein >90% of the molecular weight is distributed in the range of 10-70 kDa; Or, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 70 kDa, wherein >90% of the molecular weight is distributed in the range of 40-100 kDa; Alternatively, the small molecule hyaluronic acid is a small molecule hyaluronic acid with an average molecular weight of 24 kDa, wherein >90% of the molecular weight is distributed between 5-45 kDa.

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