New use of rutin or its hydrate, salt or derivative and injection thereof

Abstract

The present application relates to a new use of rutin or its hydrate, salt or derivative and injection thereof, and belongs to the field of biological medicine. The present application provides a use of rutin or its hydrate, salt or derivative in the preparation of a medicine for preventing or / and treating alcoholism; the medicine is an injection administration preparation, an oral administration preparation or a nasal administration preparation. The present application also provides a rutin injection prepared by including rutin in sulfobutyl-beta-cyclodextrin. The present application also provides a use of rutin or its hydrate, salt in the preparation of a dietary supplement for preventing hangover symptoms or abnormal behavior after drinking alcoholic beverages. The rutin injection or sublingual administration of the present application is safe and effective for treating alcoholism, and in addition, in the process of researching the rutin preparation, it is found that sulfobutyl-beta-cyclodextrin has a good solubilization effect on rutin, and when treating alcoholism, it can not only significantly shorten the sobering-up time, but also significantly reduce the mortality rate.

Description

Technical Field

[0001] This invention relates to novel uses of rutin or its hydrates, salts or derivatives, specifically, its use in the preparation of medicaments for the prevention and / or treatment of alcohol poisoning, belonging to the field of biomedicine. Background Technology

[0002] According to a 2018 report by the World Health Organization (WHO), more than 3 million people die annually from alcohol abuse, accounting for 5.3% of global deaths. The average daily alcohol consumption per person aged 15 and over increased from 5.5 liters in 2005 to 6.4 liters in 2016, and alcohol abuse is now a leading cause of death among men aged 15-59. For centuries, alcohol, as a potentially addictive psychoactive substance, has been widely used globally as a recreational beverage. As a hydroxyl derivative, it is highly lipid-soluble and rapidly absorbed; most alcohol is absorbed into the bloodstream, with only a small amount excreted through urine, sweat, and respiration. Excessive drinking can cause dysfunction in multiple organs and systems, often inducing stomach and liver disease, neurological and cardiovascular diseases, and cancer. In severe cases, it can even directly lead to respiratory paralysis and death. Excessive drinking has serious impacts on individual health, families, and society, posing a significant challenge to health systems worldwide.

[0003] Excessive alcohol consumption can lead to alcohol poisoning, which is classified into chronic and acute alcohol poisoning based on the different symptoms produced. Chronic alcohol poisoning can cause damage to multiple systems and organs throughout the body. The brain tissue is an important target organ of alcohol and is most vulnerable to damage. Brain injury is one of the most common and serious complications of chronic alcohol poisoning. Clinically, alcoholic encephalopathy is defined as a syndrome of mental and consciousness disorders caused by chronic damage to memory and higher mental functions related to the frontal lobe and limbic system due to long-term drinking, manifested as slowed reaction, poor concentration, memory loss, reduced spontaneous speech, tremorous delusions, hallucinations, amnesia, disorientation, ataxia, and motor disorders. Chronic alcohol poisoning can also induce apoptosis of cortical neurons in the brain (Tang Fabing, Study on apoptosis of cortical neurons induced by chronic ethanol poisoning in mice, China Pharmaceutical University, April 2010). Regarding the treatment of chronic alcohol poisoning, there are relevant literature reports, such as the protective effect of garlic polysaccharide on brain injury in mice with chronic alcohol poisoning (Wu Weiqing, Protective effect of garlic polysaccharide on brain injury in mice with chronic alcohol poisoning, Master's thesis of Jinan University, April 2012).

[0004] Acute alcoholism (AAI) refers to the accumulation of alcohol or alcoholic beverages in the body due to a single excessive intake exceeding the liver's oxidative metabolic capacity. It causes initial excitation followed by inhibition of the central nervous system, accompanied by damage to multiple systems including the liver, cardiovascular system, and cerebrovascular system. Severe poisoning can lead to respiratory and circulatory failure, ultimately resulting in death. The clinical manifestations of acute alcoholism vary from person to person and can be broadly divided into three stages: the excitation stage, the ataxia stage, and the coma stage. The pathogenesis of acute alcoholism is not fully understood, but it is related to the following factors: Because alcohol is lipid-soluble, ingested alcohol can directly affect nerve cell membranes, impacting cell function, reducing brain protein, increasing water content, and impairing myelin formation. Ethanol is a neurotropic poison that can cross the blood-brain barrier, prompting the anterior pituitary gland to release large amounts of endorphins. This enhances the activity of the endorphin system, producing morphine-like biological effects such as drowsiness and confusion. In severe cases, the cerebral cortex of the central nervous system is directly suppressed, further affecting the subcortex, medulla oblongata, and spinal cord, ultimately inhibiting vital centers and causing respiratory and circulatory failure, leading to death. Alcohol metabolism in the liver produces large amounts of substances that cause metabolic abnormalities, such as increased lactic acid and ketone body accumulation, leading to metabolic acidosis. Impaired gluconeogenesis can also cause hypoglycemia. Alcohol can affect and inhibit vitamin absorption, and impaired glucose metabolism causes abnormal changes in the function and structure of nerve tissue. When alcohol enters the body through the digestive tract, it can cause mucosal necrosis and increased gastric acid secretion, resulting in damage to the digestive tract mucosa. Long-term heavy drinking reduces the activity of digestive enzymes in the intestines, causing diarrhea due to increased intestinal fluid secretion, and eventually leading to malnutrition. Alcohol can damage the walls of pulmonary capillaries and alveoli, increasing their permeability and ultimately damaging the respiratory system. Alcohol can alter the permeability of myocardial cell membranes, causing abnormal tissue metabolism and producing toxic side effects on the heart. Therefore, acute alcohol poisoning can cause multi-system damage, leading to serious consequences (Gao Shudi, Experimental Study on the Effect of Buzuifang in Preventing Acute Alcohol Poisoning in Mice, Master's Thesis, Chengdu University of Traditional Chinese Medicine, April 2013). Clinically, the most widely accepted drug for alcohol poisoning is naloxone. Its main function is to awaken patients with alcohol poisoning and prevent death due to alcohol-induced respiratory depression, but its effect on accelerating ethanol metabolism and reducing alcohol-induced tissue damage is minimal. There are also reports in the literature of using naloxone in combination with other drugs to treat alcohol poisoning (Yao Cuie, et al., Observation on the Efficacy of Naloxone Combined with Xingnaojing in the Treatment of Acute Alcohol Poisoning, Journal of Qiqihar Medical College, Vol. 33, No. 5, 2012). Therefore, there is an urgent need to find a safe and effective drug to treat alcohol poisoning.

[0005] Rutin, also known as rutin, vitamin P, quercetin, rutin powder, rutin powder, rutin, and rutin glycoside, is clinically used primarily for capillary hemorrhage caused by increased fragility. It is also used as an adjunct treatment for hypertensive encephalopathy, cerebral hemorrhage, retinal hemorrhage, hemorrhagic purpura, acute hemorrhagic nephritis, recurrent epistaxis, traumatic pulmonary hemorrhage, and postpartum hemorrhage. Studies have reported that rutin has anti-inflammatory effects (Liu Xin, PK-PD model study of the anti-inflammatory effect of rutin in mice with lipopolysaccharide-induced acute lung injury, Shenyang Agricultural University, December 2020); rutin also has anti-hypoxic effects (Yao Juan, Study on the anti-hypoxic effect and molecular mechanism of rutin, Lanzhou University, April 2012, application number 201110392838.6). Literature reports that rutin can be used to treat psoriasis and tumors (liver cancer, breast cancer, ovarian cancer, and colon cancer). Patent application number 200910216333.7, entitled "New Uses of Rutin," discloses the use of rutin in the preparation of hepatoprotective drugs. Pharmaceutical experiments have demonstrated that rutin can effectively reduce cryopreservation-reperfusion injury in liver transplant recipients and has a protective effect on the liver. However, some literature reports side effects such as liver damage and hematopoietic dysfunction. Furthermore, rutin has unique solubility; it is poorly soluble in water, slightly soluble in ethanol, acetone, and ethyl acetate, and almost insoluble in chloroform, benzene, disulfides, and petroleum ether. Due to its poor water and lipid solubility, the only currently available marketed dosage form of rutin is oral tablets. The usual adult dose is 20-40 mg (1-2 tablets) three times daily, limiting the clinical use of rutin.

[0006] There are currently no reports of rutin being used to treat alcohol poisoning, nor are there any reports of new methods of administering rutin. Summary of the Invention

[0007] To address the aforementioned problems, the present invention provides a new use for rutin, and another technical solution of the present invention is to provide a rutin injection.

[0008] This invention provides the use of rutin or its hydrates, salts or derivatives in the preparation of medicaments for the prevention and / or treatment of alcohol poisoning.

[0009] The alcohol poisoning mentioned above includes acute alcohol poisoning and chronic alcohol poisoning.

[0010] The medication mentioned is for the prevention and / or treatment of hangovers or post-drinking disorders.

[0011] The drug is an injectable preparation, an oral preparation, or a nasal preparation; preferably, the injectable preparation is an injection solution or a lyophilized powder for injection; and the oral preparation is a sublingual preparation.

[0012] More preferably, the sublingual administration preparation is a sublingual tablet, sublingual drop, sublingual spray, sublingual liquid, or sublingual film.

[0013] The human dosage of the injectable formulation is 0.03-3 mg / kg; the human dosage of the oral formulation is 0.09-9 mg / kg.

[0014] The present invention also provides a rutin injection, wherein the injection comprises sulfobutyl-β-cyclodextrin and rutin; wherein the weight ratio of rutin to sulfobutyl-β-cyclodextrin is:

[0015] 1 part rutin, 9-50 parts sulfobutyl-β-cyclodextrin.

[0016] The preparation process of this injection is as follows:

[0017] a. Weigh the raw materials rutin and sulfobutyl-β-cyclodextrin, and their weight ratio is as follows:

[0018] 1 part rutin, 9-50 parts sulfobutyl-β-cyclodextrin;

[0019] b. Add the raw materials and excipients from step a to physiological saline, heat to 70°C, stir and maintain for 15-25 minutes, then cool to room temperature to obtain a clear solution of rutin inclusion complex. Add pharmaceutically acceptable excipients, sterilize, and obtain the injection solution. Further freeze-dry the injection solution to obtain freeze-dried powder for injection.

[0020] This invention provides a safe and effective method for treating alcohol poisoning via injection or sublingual administration of rutin. During the research on rutin preparations, it was discovered that sulfobutyl-β-cyclodextrin has a good solubilizing effect on rutin. Furthermore, in the treatment of alcohol poisoning, it can not only significantly shorten the time to sobriety but also significantly reduce the mortality rate. It can be used to prepare drugs with a wide range of applications to meet clinical needs. Attached Figure Description

[0021] Figure 1 Effect of different mass ratios of rutin and sulfobutyl-β-cyclodextrin on the solubilization of rutin (mass ratios were 1:50, 1:30, 1:20, 1:12, 1:9, 1:7, and 1:5, respectively);

[0022] Figure 2 Results of rutin intravenous injection efficacy test;

[0023] Figure 3 The effect of intravenous rutin injection on blood alcohol concentration;

[0024] Figure 4 Results of a pharmacodynamic test of rutin administered sublingually. Detailed Implementation

[0025] The instruments and reagents used in this invention:

[0026] Thermostatic water bath magnetic stirrer; CPA225D electronic balance (Sartorius, Germany); ModulyoD freeze dryer (Thermo Fisher Scientific, USA); 7890B gas chromatograph (Agilent Technologies, USA); rutin (Shanghai Yuanye Biotechnology Co., Ltd., 98%); rutin hydrate (Aladdin, 98%); naloxone hydrochloride (Aladdin, 99% purity); α-cyclodextrin, γ-cyclodextrin and β-cyclodextrin were purchased from Wacker, Germany; hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin were purchased from Zibo Qianhui Biotechnology Co., Ltd.; PEARLITOL® Flash (mannitol starch co-processed product) was purchased from Roquette, France.

[0027] Example 1 Preparation of Rutin Inclusion Complex

[0028] Add 15 mg of rutin and 190 mg of sulfobutyl-β-cyclodextrin to 5 ml of physiological saline, heat to 70 °C, stir and maintain for 20 min, then cool to room temperature to obtain a clear solution of rutin inclusion complex.

[0029] Example 2: Lyophilization of Rutin Inclusion Complex

[0030] Add 15 mg of rutin and 190 mg of sulfobutyl-β-cyclodextrin to 5 ml of physiological saline, heat to 70 °C, stir and maintain for 20 min, then cool to room temperature to obtain a clear solution of the rutin inclusion complex. Add 10% mannitol to the above solution and freeze-dry to obtain the lyophilized powder of the rutin inclusion complex.

[0031] Example 3 Preparation of Rutin Inclusion Complex

[0032] Rutin and sulfobutyl-β-cyclodextrin at different mass ratios (1:5, 1:7, 1:9, 1:12, 1:20, 1:30, 1:50) were added to 5 ml of physiological saline, heated to 70°C, stirred and maintained for 20 min, then cooled to room temperature, and the state of the solution was observed. The results showed that clear solutions were obtained when the mass ratio of rutin to sulfobutyl-β-cyclodextrin was 1:9-50. However, obvious precipitation occurred when the mass ratio of rutin to sulfobutyl-β-cyclodextrin was 1:7 and 1:5. See the results below. Figure 1 .

[0033] Example 4: Preparation of Rutin Sublingual Tablets

[0034] Mix 1.5g of rutin hydrate, 18.5g of mannitol starch co-treated product, 220mg of aspartame, and 260mg of magnesium stearate evenly, compress directly into tablets, and make 100 tablets.

[0035] Comparative Example 1: Preparation of Rutin Inclusion Complex

[0036] 190 mg of α-cyclodextrin and 15 mg of rutin were added to 5 ml of physiological saline, heated to 70 °C, stirred and maintained for 20 min, and then cooled to room temperature. Rutin inclusion complexes were prepared using the same amount of β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and γ-cyclodextrin instead of α-cyclodextrin, following the same method. The results showed that a clear solution without precipitate could not be obtained using α-cyclodextrin, β-cyclodextrin, or hydroxypropyl-β-cyclodextrin.

[0037] The above experiments demonstrate that the selection of cyclic dextrins in rutin inclusion complexes is specific; only sulfobutyl-β-cyclic dextrins can achieve inclusion and solve the rutin solubility problem.

[0038] The following specific pharmacodynamic tests demonstrate the beneficial effects of this invention.

[0039] Experimental Example 1: Pharmacodynamic Study of Rutin Intravenous Injection in this Invention

[0040] Naloxone hydrochloride was dissolved in physiological saline to prepare a naloxone solution. Mice were randomly divided into 9 groups of 11 mice each. Each mouse was given a 50% ethanol solution (0.1 ml / 10 g) via intraperitoneal injection. Ten minutes after the righting reflex disappeared, each group of mice was given an equal volume of physiological saline (saline group), a 38 mg / ml sulfobutyl-β-cyclodextrin saline solution (blank cyclodextrin group), a naloxone solution (naloxone dose of 3 mg / kg, naloxone group), or different doses of rutin inclusion complex solution prepared according to Example 1 via tail vein injection. The mice were administered rutin solutions (rutin doses of 3 mg / kg, 1.5 mg / kg, 0.75 mg / kg, 0.3 mg / kg, and 0.1 mg / kg, respectively, and were designated as the rutin group (3 mg / kg), rutin group (1.5 mg / kg), rutin group (0.75 mg / kg), rutin group (0.3 mg / kg), and rutin group (0.1 mg / kg)). The last group of mice was administered the rutin inclusion complex solution prepared in Example 1 (rutin dose of 3 mg / kg) by gavage, and was designated as the rutin gavage group (3 mg / kg). The time and rate of recovery of the righting reflex within 2 hours (the mouse was considered to have recovered the righting reflex if it could right itself twice within 30 seconds) were recorded, and the mortality rate of each group of mice within 4 hours was also recorded.

[0041] The results showed that the percentages of mice recovering their righting reflex within 2 hours in the saline group, blank cyclodextrin group, naloxone group, rutin group (3 mg / kg), rutin group (1.5 mg / kg), rutin group (0.75 mg / kg), rutin group (0.3 mg / kg), rutin group (0.1 mg / kg), and rutin gavage group (3 mg / kg) were 45.5%, 45.5%, 63.6%, 81.8%, 81.8%, 72.7%, 63.6%, 54.5%, and 45%, respectively. 0.5%; Within a 4-hour observation period, the mortality rates of mice in the saline group, blank cyclodextrin group, naloxone group, rutin group (3 mg / kg), rutin group (1.5 mg / kg), rutin group (0.75 mg / kg), rutin group (0.3 mg / kg), rutin group (0.1 mg / kg), and rutin gavage group (3 mg / kg) were 54.5%, 54.5%, 45.5%, 18.2%, 27.3%, 36.4%, 45.5%, 54.5%, and 54.5%, respectively. The time for mice in each group to recover their righting reflex was as follows: Figure 2 As shown, naloxone can rapidly awaken alcohol-poisoned mice, but the mortality rate in this group was high during the observation period (some mice in this group lost their righting reflex again after awakening, thus dying within the 4-hour observation period). After intravenous treatment with different doses of rutin, compared to the saline control group, the time to recovery of the righting reflex in alcohol-poisoned mice (especially those with rutin doses ≥0.3 mg / kg) was reduced to varying degrees, and the mortality rate of these rutin-treated mice was lower than that of the saline group during the observation period; however, no similar effect was observed when rutin was administered by gavage. These results fully demonstrate that the rutin injection of the present invention has a significant therapeutic effect on alcohol poisoning.

[0042] Experimental Example 2: Effect of intravenous rutin injection of the present invention on blood alcohol concentration

[0043] Rats were randomly divided into four groups of five each. Each rat was administered a 50% ethanol solution (0.55 ml / 100 g) via intraperitoneal injection. Five minutes after ethanol administration, each group of rats was administered an equal volume of physiological saline (saline group), a 38 mg / ml sulfobutyl-β-cyclodextrin saline solution (blank cyclodextrin group), naloxone solution (2 mg / kg naloxone group), or a rutin inclusion complex solution prepared according to Example 1 (2 mg / kg rutin group) via tail vein injection. Blood samples were collected via ocular vein at 10, 30, 60, 120, 240, and 360 minutes after administration, and the ethanol concentration in the blood was measured using headspace analysis.

[0044] The results showed that the blood alcohol concentration in the blank cyclodextrin group was almost the same as that in the saline group; compared with the saline group, the blood alcohol concentration in the naloxone group tended to decrease, but the difference was not statistically significant; the blood alcohol concentration in the rutin group was significantly lower than that in the saline group. These results indicate that the rutin of the present invention can accelerate the metabolism of blood alcohol (see...). Figure 3 ).

[0045] Experimental Example 3: Pharmacodynamic Test of Sublingual Administration of Rutin in this Invention

[0046] Mice were randomly divided into 4 groups of 12 each. Each mouse was given a 50% ethanol solution (0.1 ml / 10 g) via intraperitoneal injection. After the righting reflex disappeared for 10 minutes, each group of mice was administered an equal volume of physiological saline (saline group), a 38 mg / ml sulfobutyl-β-cyclodextrin saline solution (blank cyclodextrin group), a rutin inclusion complex solution prepared in Example 1 as a sublingual drop (rutin dose 9 mg / kg, rutin solution group), or a lyophilized tablet prepared according to the formulation in Example 2 (rutin dose 9 mg / kg, lyophilized rutin group). The time to righting reflex recovery (the mouse was considered to have recovered righting reflex if it could right itself twice within 30 seconds) and the rate of recovery were recorded within 2 hours. The mortality rate of each group of mice within 4 hours was also recorded.

[0047] The results showed that the rates of righting reflex recovery within 2 hours in the saline group, blank cyclodextrin group, rutin solution group, and freeze-dried rutin group were 41.7%, 50.0%, 75.0%, and 66.7%, respectively; the mortality rates within 4 hours in the saline group, blank cyclodextrin group, rutin solution group, and freeze-dried rutin group were 58.3%, 50%, 25.0%, and 25.0%, respectively. The time for righting reflex recovery in each group is shown in the figure below. Figure 4 As shown, mice in the saline group and the blank cyclodextrin group had longer recovery times and higher mortality rates during the observation period. However, after treatment with sublingual rutin solution and lyophilized rutin, compared to the control group, the time for alcohol-poisoned mice to regain their righting reflex was significantly reduced, and the mortality rate during the observation period was significantly lower in the latter group. These results fully demonstrate that the rutin of this invention can treat alcohol poisoning via sublingual administration.

[0048] Test Example 4: Test on the prevention of alcohol poisoning by rutin in this invention

[0049] Mice were randomly divided into two groups of 15 each. One group (control group) received an intraperitoneal injection of a 50% ethanol solution (0.1 ml / 10 g) per mouse; the other group (rutin group) received an intraperitoneal injection of a 50% ethanol solution containing rutin (0.1 ml / 10 g, rutin dose 3 mg / kg). After injection, the time and percentage of mice losing their righting reflex were recorded.

[0050] The results showed that all mice in the control group exhibited loss of the righting reflex, with an average disappearance time of only 2.5 ± 1.2 min. In contrast, 66.7% of the mice in the rutin group did not show loss of the righting reflex during the 2-hour observation period, and the remaining 33.3% of mice in this group showed loss of the righting reflex with an average time of 10.2 ± 6.5 min. These results indicate that the rutin of this invention can prevent alcohol poisoning.

[0051] Experimental Example 5: Clinical Trial of Sublingual Administration of Rutin in this Invention

[0052] Subjects were given an alcoholic beverage containing ethanol until their blood ethanol concentration reached 300 mg / 100 mL. They were then given 500 mL of water and immediately afterwards administered a sublingual tablet containing 15 mg of rutin. Afterward, the subjects were put to sleep. Hangover symptoms were recorded after 8 hours of sleep. As a comparison, the same trial was conducted as a placebo with a sublingual tablet containing no rutin. Each group consisted of 16 subjects.

[0053] The results showed that among the subjects taking the placebo, 11 subjects experienced significant hangover symptoms such as headache and fatigue, 3 subjects experienced mild dizziness and fatigue, and 2 subjects had no typical hangover symptoms. Among the subjects taking sublingual tablets containing rutin, only 3 subjects experienced significant hangover symptoms such as headache and fatigue, 5 subjects experienced mild dizziness and fatigue, and 8 subjects did not have any typical hangover symptoms usually associated with drinking alcoholic beverages.

Claims

1. Use of rutin injection in the preparation of medicaments for the prevention and / or treatment of alcohol poisoning; said injection comprising sulfobutyl-β-cyclodextrin and rutin; wherein, The weight ratio of rutin to sulfobutyl-β-cyclodextrin is: 1 part rutin, 9-50 parts sulfobutyl-β-cyclodextrin.

2. The use according to claim 1, characterized in that: The preparation process of this injection is as follows: a. Weigh the raw materials rutin and sulfobutyl-β-cyclodextrin, and their weight ratio is as follows: 1 part rutin, 9-50 parts sulfobutyl-β-cyclodextrin; b. Add the raw materials and excipients from step a to physiological saline, heat to 70°C, stir and maintain for 15-25 minutes, then cool to room temperature to obtain a clear solution of rutin inclusion complex. Add pharmaceutically acceptable excipients, sterilize, and obtain the injection solution. Further freeze-dry the injection solution to obtain freeze-dried powder for injection.

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