CN100464760C

CN100464760C - Use of cocklebur fruit total glucoside extract in preparing anti inflammation reaction product

Info

Publication number: CN100464760C
Application number: CNB2005100246939A
Authority: CN
Inventors: 秦路平; 韩婷; 张巧艳; 黄宝康; 郑汉臣
Original assignee: Second Military Medical University SMMU
Current assignee: Second Military Medical University SMMU
Priority date: 2005-03-28
Filing date: 2005-03-28
Publication date: 2009-03-04
Anticipated expiration: 2025-03-28
Also published as: CN1686254A

Abstract

An application of the xanthinoside, which is prepared from xanthium fruit through extracting, separating and purifying, in preparing the medicine for relieving inflammation, relaxing pain and treating rhinitis and nasosinusitis is disclosed.

Description

Application of cocklebur fruit total glycoside extract in preparing anti-inflammatory reaction product

Technical Field

The invention relates to the technical field of medicines, foods and beverages, in particular to application of a traditional Chinese medicine extract, and more particularly to application of a traditional Chinese medicine xanthium sibiricum total glycoside extract in preparation of an anti-inflammatory reaction product.

Background

General study of Xanthium strumarium

1. Plant resource and application profiles

Fructus Xanthii (Fructus Xanthii) is a dry mature fruit with involucre of Xanthium sibiricum Bates (Xanthium sibirium Batr) of Compositae, also called Solanum torvum Linne, Angelica sinensis Diels, furuncle grass, Aralia chinensis, viscous sunflower, Fructus Xanthii and so on, is a Chinese pharmacopoeia variety, produced in most areas of China, and has abundant resources. The traditional Chinese medicine considers that the medicine is pungent and bitter in taste and warm in nature, enters lung and liver meridians, and has the effects of sweating, dispelling rheumatism, dredging nasal orifices and relieving pain. The traditional Chinese medicine is used for treating wind-cold headache, nasosinusitis, rhinorrhea, rubella pruritus, damp arthralgia, contracture and the like (pharmacopoeia committee of the people 'S republic of China, pharmacopoeia of the people' S republic of China [ S ]. part one, Beijing: chemical industry publishing company, 2000: 128); it is clinically used for treating acute and chronic rhinitis, nasosinusitis, tracheitis, lumbago and skelalgia, rubella, scabies and other skin diseases (national administration of traditional Chinese medicine [ M ]. seventh volume, Shanghai: Shanghai science and technology publishing house, 1999: 1013-1016).

2. Chemical composition

Research shows that the chemical components contained in the xanthium plants are mainly sesquiterpene lactone, volatile oil and water-soluble glycosides, which are as follows:

(1) sesquiterpene lactone compounds xanthium plants mainly contain guaiane type and guaiane type lactone compounds, and mainly comprise xanthinin (cryptoxanthin), xanthinin (xanthinin, a stereoisomer of xanthinin), xanthatin (xanthatin), derivatives thereof and the like. There are also xanthinol (xanthanol), xanthinol, and xanthinol, as well as xanthinol A, xanthinol B in X.canadens, ziniolide in X.catharticum, lasidiol p-methoybenzoate, and pulgiolide A, pulgiolide B in X.puresens (Salinas-A, De-Ruiz-RE, Ruiz-SO.Sterols, flavanoids and sesterpending lactic acid from xanthinol (Aspergillus), Acta Farm Borarenense, 1998, 17: 297; and Kis-I, Racz-G.Glosyl. glycoside contents of xanthenols from Xihinum X.physiognosis, Famili-G.18, and Xanthomonas-II, and Xizhi.35: Xilun-G.8. xanthinol and Ophiol. 1990, and Xizhi-X.8. xanthinol (Axib. 35, Axix.8, and Xizhi. 9: Xizhi).

(2) Volatile oil Ahijja-MM, etc. volatile oil from Xanthium X.STRUMARIUM leaves (Ahijja-MM, Nigam-SS.chemical evolution of the essential oil from the leaves of Xanthiunstrucrium (Linn.). Flavoner Ind, 1970, 1: 627), which was found to contain d-limonene (d-limonene, 35%), cymene (p-cymene, 5%), beta-caryophyllene (6%), terpinolene (7.3%), 1-alpha-ionone (l-alpha-ionone, 10.9%), d-homoprenol (d-carveol, 25%) and alpha-pinene (alpha-pinene, 0.4%). Taher-HA et al analyzed the volatile oil content of X.canonillesii (Taher-HA, Ubiergo-GO, Talenti-ECJ.Constitutens of the invasive oil of Xanthiun canonillesii.J. Nat Prod, 1985, 48: 857.), which contained 55.2% monoterpene hydrocarbons, 14.2% oxidized monoterpenes, and 11.3% sesquiterpene hydrocarbons. Guoia red, etc. separated 17 chemical components from the volatile oil of Xanthium sibiricum and calculated the relative percentage (Guoia red, plum fruit, shiny, etc. study of the volatile oil in Xanthium sibiricum. Chinese materia medica journal, 1994, 19 (4): 235).

(3) A substance (temporarily named as AA2) with glycoside property is separated from the water extract of cocklebur fruit in the last 60 th century, such as water-soluble glycoside Song Zhen Yu, and may be the main toxic component of cocklebur fruit (Song Zhen Yu, Zhang Ling Yun, Xie Ming Zhi, etc. the toxic component of cocklebur fruit and its pharmacological action, reports in pharmacy 1962, 9 (11): 678). Two water-soluble toxic kaurene glycoside components (MacLeod-JK, Moeller-PD, Franke-FP. two toxin glycosides from the burrs of Xanthium strung.J. Nat Prod, 1990, 53: 451) extracted from the thorn fruit of Xanthium strumarium X.pungens by MacLeod-JK, etc. are identified by two-dimensional nuclear magnetic resonance and mass spectrometry techniques to determine the derivative with the structure of carboxyl atractyloside (carboxyactylide). In addition, the same water-soluble toxic glycoside is extracted from fructus Xanthii and Mongolian fructus Xanthii and identified as atracyloside (a toxic component in Mongolian fructus Xanthii kernel, Chinese herbal medicine 1983, 14 (12): 1).

(4) Other xanthium sibiricum contains xanthium glycoside (beta-sitosterol glucoside), 8- (delta 3-isopentenyl) -5, 7, 3 ', 4' -tetrahydroxyflavone, caffeic acid and 1, 4-dicaffeoylquinic acid. In addition, alkaloid, tannin, chalcone derivative, triterpenes, glucose, fructose, amino acid, tartaric acid, etc. can be added.

3. Pharmacological Activity

(1) The cocklebur fruit decoction has strong inhibiting effect on staphylococcus aureus, bacillus anthracis, pneumococcus, streptococcus B, diphtheria bacillus and other microbes in vitro. The acetone or ethanol extract of Xanthium strumarium has antifungal effect on Trichophyton rubrum, and the water extract has antifungal effect on Trichophyton violaceum. Zhang Zheng is equal to that the research in 1988 proves that the direct inhibition rate of the cocklebur fruit decoction on the hepatitis B virus DNA polysaccharide in vitro is 25-50%, which shows that the cocklebur fruit decoction has the effect of resisting the hepatitis B virus (Zhang Zheng, Dong, Dushao wealth, etc. 60 experimental researches on the anti-hepatitis B virus of Chinese herbal medicines, Beijing university of medical science, 1988, 20 (3): 211).

(2) The decoction has effect in inhibiting heart rate of frog and guinea pig, reducing heart contraction force, dilating blood vessel of rabbit ear, dilating blood vessel of frog, and contracting blood vessel of frog. Glycoside AA2 has mild hypotensive effect on rat and can enhance vascular permeability (Song Zhen Yu, Zhang Ling Yun, Xie Ming Zhi, etc., toxic component of Xanthium sibiricum and its pharmacological action, pharmaceutical science, 1962, 9 (11): 678).

(3) The fructus Xanthii extract (equivalent to 0.2g/ml crude drug) can remarkably prolong the time of bovine thrombin coagulating human fibrinogen, and has obvious antithrombin effect. The fructus Xanthii methanol extract can rapidly recover the reduction of cholesterol and triglyceride of rabbit caused by fasting, and can increase phospholipid content to a certain extent.

(4) The cocklebur fruit has obvious inhibition effect on the cellular immunity and the humoral immunity of the C57/BL pure mouse. Xanthium strumarium reduces the number of helper T cells (abbreviated as TH) and suppressor T cells (abbreviated as TS) and decreases the TH/TS ratio. Xanthium sibiricum has a significant reducing effect on beta-endorphin in the hypothalamus and plasma. In addition, Xanthium strumarium can reduce the activity of interleukin-2 (IL-2 for short) and the content of IL-2 receptor, and can obviously reduce the release of histamine in cells, which is one of the mechanisms by which Xanthium strumarium can be used for treating allergic diseases (Wangloimei, Fu Hui, Zhou Chi, Gou Qi Zi, Bai Zhu, xi Xin, Xanthium strumarium has an effect on the expression of interleukin-2 receptor. J. Chinese J. Clematis & Pharmacology 2000, 9 (3): 172).

(5) Antioxidation effect of fructus Xanthii was studied in 1994, and it was shown that fructus Xanthii decoction (0.5 g crude drug/one herbal drug) was infused into the stomach 1 time per day for 10 days continuously, which can effectively reduce lipid peroxidation, reduce lipid peroxidation in tissues (LPO for short), and improve superoxide dismutase (SOD) activity, indicating that fructus Xanthii can enhance the scavenging ability of organism to free radicals, and reduce the damage of free radicals to organism (fructus Xanthii, herba asari, fructus Lycii, rhizoma Atractylodis Macrocephalae influence on free radical metabolism in mouse tissue. Chinese medicinal information, 1994, (2): 48).

(6) Antiinflammatory and analgesic effects Liu Qing increase et al, 1988, have described pharmacological activity of Xanthium sibiricum extract (Liu Qing increase, Wang jin lan. some progress on the study of pharmacological effects of Chinese herbs in recent years. Chinese pharmacology and clinic, 1988, 4 (2): 50). The cocklebur fruit methanol extract is injected into the abdominal cavity at 250mg/kg, and the inhibition rate of the rat carrageenan podoma is 30-60 percent; the inhibition rate of 1000mg/kg subcutaneous injection on the acetic acid writhing reaction of mice is 10-30%, which shows that the product has certain anti-inflammatory and analgesic effects. The modified fructus xanthil pills have good anti-inflammatory effect and comprise the following components: can obviously reduce the infiltration amount of evans blue in the abdominal cavity of the mouse caused by acetic acid and can obviously reduce the swelling degree of ear inflammation of the mouse caused by dimethylbenzene. Has good analgesic effect, and can significantly prolong the time of writhing reaction of mice caused by acetic acid (Lemna minor, plum red. xanthium fruit research progress. Chinese herbal medicine 1999, 30 (8): 634).

(7) Other researches show that the external inhibition rate of the cocklebur fruit aqueous extract on cervical cancer cells reaches 50 to 70 percent; the fructus Xanthii extract has different degrees of inhibition effects on angiotensin receptor, beta-hydroxy-beta-methylglutaric acid coenzyme A (HMG-Co-A for short), calcium channel blocker receptor, cholecystokinin and the like.

4. Clinical applications of Single prescription and Compound (Li hong, consummation, pharmacological action and clinical research progress of Xanthium sibiricum and Compound preparation, university of Shanxi medical science, 2004, 35 (3): 313-

(1) Treating rhinopathy

Treating rhinitis and chronic rhinitis is a common disease and frequently encountered disease. According to the traditional fructus xanthil powder, reduction cutting is carried out on the basis of the traditional fructus xanthil powder, 96 patients are randomly divided into a treatment group and a control group, 1 month is 1 treatment course, no significant difference exists between the two groups through clinical observation, and the modified fructus xanthil pill is close to and slightly superior to the rhinitis pill in treating chronic rhinitis and rhinitis.

The scutellaria decoction and the siberian cocklebur fruit powder for treating the nasosinusitis are used for treating the chronic nasosinusitis, the total effective rate is 94.4 percent by clinical observation of 108 cases, and the traditional Chinese medicine can obviously take effect after one treatment course of general medication. The cocklebur fruit mixture and the paracetamol are used together to treat the chronic nasosinusitis of children, and the effect is obvious. The cocklebur fruit is baked into brown color and then ground into powder or the powder is made into honeyed pills to treat allergic rhinitis, and symptoms of most patients disappear or are improved or have less attack. Also, 67 cases of chronic nasosinusitis can be treated by using the ancient formula siberian cocklebur fruit powder, myrobalam, earthworm and the like, 42 cases are cured, 20 cases are improved, 5 cases are not effective, and the total effective rate is 92.54%.

The traditional Chinese medicine for treating nasosinusitis classifies the acute purulent nasosinusitis as nasosinusitis. 200 cases of acute purulent nasosinusitis are treated by cocklebur fruit mixture, 100 cases of treatment group take the cocklebur fruit mixture, and 100 cases of control group select penicillin, compound sulfamethoxazole and the like for symptomatic treatment. The clinical treatment results show that the total effective rates of the treatment group and the control group are respectively 99 percent and 95 percent (P is more than 0.05), and no obvious difference exists. Clinically, the decoction of cocklebur and pungent is also commonly used for treating nasosinusitis and allergic rhinitis, and reports of the powder of cocklebur fruit of Jade Screen for treating allergic rhinitis and the like show the contribution of cocklebur fruit in treating rhinopathy.

(2) The cocklebur fruit for treating cancer is prepared into a compound preparation in different formula forms, and multiple cases of nasopharyngeal carcinoma and nasal cavity cancer are effectively cured clinically; 3 cases of penile cancer are treated and clinically cured by matching with an operation; and has certain curative effect on intracranial tumor, malignant tumor of nervous system, brain tumor, etc.

(3) The literature reports about treating odontopathy that 98 cases of intractable toothache are treated by ancestral single prescription, and the curative effect is satisfactory. Also, 15g of siberian cocklebur fruit and figwort root are decocted with water respectively for oral administration, and the report shows that the medicine has curative effect on toothache.

(4) The dermatitis treatment comprises 25 cases (65.8 percent) of 38 dermatitis patients, 10 cases (26.8 percent) of significant effect, 3 cases (7.9 percent) of ineffective effect and 92.1 percent of total effective rate.

(5) The literature reports that 30 outpatients with upper respiratory tract infection are treated by using the modified xanthium sibiricum, 26 cases (86.7%) are cured, 3 cases (10.0%) are improved, and 1 case is invalid; the total effective rate reaches 96.7 percent. The medicine is taken for 2 doses at minimum and 5 doses at maximum.

(6) About 50g to 70g of xanthium sibiricum for treating diarrhea is added with 3000ml of clear water to be soaked for 30min, then the mixture is boiled by strong fire and then is decocted by slow fire for 15min, the liquid medicine is filtered out, the temperature is cooled to 35 ℃ to 38 ℃, and the warm liquid is used for dipping or soaking the crus and feet 3 times per day; 46 cases are cured, the cure rate is 95.77%, 1 case is effective, the effective rate is 2.15%, and 1 case is ineffective (accounting for 2.15%).

(7) The literature reports that clinically, the xanthium sibiricum powder is matched with the antibacterial dry powder to treat the purulent otitis media and the pure purulent otitis media has good treatment effect.

(8) 6000g of typhoid cocklebur is treated, the typhoid cocklebur is soaked in clear water and heated and boiled for 1h, decoction liquid is filtered out, the decoction liquid is continuously decocted for 3 times by the same method, the decoction liquid for 3 times is mixed, filtered, concentrated by slow fire to 2000ml, 10g of nipagin is added, the mixture is evenly shaken, 15 typhoid fever cases are effectively treated, the defervescence time is 10h fastest, 7 cases of positive blood, feces and bile typhoid bacillus are treated, and all the typhoid bacillus turns negative after treatment.

(9) 100g of fresh cocklebur for treating dysentery is cleaned and mashed, water is added for decocting for 15min, dregs are removed, 2-3 eggs are added, the mixture is boiled in liquid medicine, the liquid medicine and the eggs are taken together before onset, the eggs are not healed for 1 time, the mixture can be taken by the method, and the curative effect of treating the dysentery is good.

(10) 1 dose of decoction is taken by 2 times per day for treating sore, furuncle, cocklebur, burdock, raw rhubarb and the like; 4-8 continuous doses can obtain good effect for treating more than 30 cases of sores and furuncles.

(11) Other clinical applications also report that xanthium sibiricum and compound preparations thereof are used for treating lumbocrural pain, parotitis and lower limb ulcer clinically, and all the clinical applications have a certain curative effect.

Fructus xanthil is used singly or combined with other traditional Chinese medicines clinically, is used for treating common and difficult and complicated nasal diseases such as acute and chronic rhinitis, nasosinusitis, allergic rhinitis and the like, and achieves better curative effects, such as pungent nasal drop, fructus xanthil powder, xanthil-angelica decoction, pungent fish and angelica decoction and the like, which are famous prescriptions for clearing stagnated heat, eliminating dampness and turbidity and opening nasal orifices, and have been applied clinically for many years and have better curative effects. However, due to the fact that the research on chemical components is not deep, the pharmacological screening is insufficient, the active site and the active components are not clear, and no feasible quality standard exists, and the further development and utilization of the cocklebur fruit are limited.

Research progress of sinusitis

Sinusitis (sinusitis) is an acute or chronic inflammation of the sinuses that occurs as a result of bacterial invasion into the sinuses, destroying the sinus tissue. The site of attack is most frequently maxillary sinusitis, followed by ethmoid sinusitis, frontal sinusitis and sphenoid sinusitis, and if all sinuses are involved, it is called sinusitis (pansinusitis).

Acute and chronic sinusitis is a common disease and frequently encountered disease, and 3700 thousands of chronic sinusitis patients in the United states only are reported in the literature; some U.S. researchers believe that patients with global sinusitis account for approximately 14% of the world's total population (about eight hundred and four million people), and that the number of cases of sinusitis is rapidly increasing year by year. In China, the incidence of sinusitis fluctuates between 10.7% and 44.5% due to different regions. According to the statistics of clinical data, the incidence of chronic nasosinusitis accounts for 13.02% of the initial diagnosis patients of the otorhinolaryngology department. Particularly in southern China, because of humid climate, the incidence rate of nasosinusitis is very high, and about more than 1/3 of the initial patients in otorhinolaryngology outpatient clinic of Shanghai Changhai hospital are various nasosinusitis patients, wherein the patients are 76.0% under 30 years old, 6.4% between 40-50 years old and 1% or 2% after 50 years old.

Due to sinus ostia stenosis, unfavorable excretion and allergic constitution, the disease usually attacks repeatedly and is difficult to cure. Moreover, the disease has long course of disease and is often accompanied with symptoms such as headache, dizziness, hypomnesis and the like, thereby bringing great adverse effect to the study, work and life of patients.

In addition, after nasosinusitis occurs, inflammation can spread and invade adjacent tissues, osteomyelitis, orbital cellulitis, pia meningitis and cerebral abscess are easily caused, severe purulent nasosinusitis can even cause septicemia, and the life of a patient is directly endangered. Repeated stimulation of chronic sinusitis has also been shown to cause cancer.

Modern studies have shown that chronic rhinosinusitis is the result of a combination of factors, but many of them are still unknown and their pathogenesis is not fully elucidated. The study of Chinese scholars on the pathogenic factors mainly focuses on the following aspects: bacterial or viral infection; secondly, the sinus-oral-nasal passage complex (OMC for short) is blocked; ③ mucociliary dysfunction; fourthly, immune dysfunction and the like.

The pathogenic bacteria of sinusitis are pyococcus such as Diplococcus pneumoniae, hemolytic streptococcus, staphylococcus, etc., and Bacillus such as Bacillus influenzae, Bacillus proteus, etc. Clinically, the occurrence of nasosinusitis is mostly caused by mixed infection of two pathogenic bacteria. Bacterial invasion impairs host resistance, reduces nasal mucociliary function, and even paralyzes mucosal defense mechanisms.

Recent studies have shown that obstruction of the osteomeatal complex by various causes leading to obstruction of ventilation and drainage is the main cause of delayed and recurrent episodes of sinusitis. Meanwhile, the links do not exist in isolation, but influence and interact with each other, so that the persistent inflammatory reaction and the irreversible pathological change of the mucosa of the paranasal sinus are caused.

The sinusitis has high morbidity, is easy to cause serious complications due to the spread of inflammation, is easy to repeatedly attack and is difficult to heal after a long time. At present, the western medicines which are commonly used clinically mainly comprise antibiotics, decongestants, phlegm-resolving agents, analgesics, steroids, nasal sprays and the like, are only suitable for patients with mild symptoms and early stages and patients before operations, can only improve symptoms to different degrees, and cannot radically cure nasosinusitis. About 2/3 sinusitis patients have different degrees of side effects after external application or taking of western medicines, such as ephedrine nose drops which are the most common, and long-term repeated use can cause nasal cavity vasomotor dysfunction, olfactory nerve ending degeneration and finally form rhinitis medicamentosa; abuse of antibiotics can cause pathogenic bacteria to develop drug resistance, and can also disrupt the balance of normal flora, causing imbalance and adverse consequences. In summary, although the market has a lot of drugs for treating nasosinusitis, the clinical application of the drugs is limited by the defects of unstable curative effect, side effects of different degrees and the like.

(III) Advances in the study of inflammation

Exogenous and endogenous injury factors can cause various kinds of injury pathological changes of cells, and at the same time, a series of complex reactions occur locally and systemically in the body to limit and eliminate the injury factors, eliminate and absorb necrotic tissues and cells, and repair injuries, namely defensive reactions of the body, namely inflammation (inflammation).

Inflammation is characterized clinically by redness, swelling, pain and dysfunction, and systemic reactions caused by inflammation include fever and increased peripheral blood leukocyte counts. Inflammation is originally the first line of defense of the body against viral infection and is a protection for the body. However, when the degree of inflammation is severe or it develops into chronic inflammation, it may cause a series of other diseases such as heart attack, colon cancer and presenile dementia.

At present, the anti-inflammatory analgesics commonly used in clinic are mainly non-steroidal anti-inflammatory drugs (NSAIDs). NSAIDs have been widely used for the analgesia of patients with rheumatoid and non-rheumatic diseases, including abdominal pain and menstrual pain such as acute postoperative pain, chronic pain, biliary colic and renal colic. The traditional non-steroidal anti-inflammatory drugs inevitably have damage to the gastrointestinal tract, blood coagulation function and renal function due to the increase of the dosage, the course of treatment and the age of patients, and limit the application range of the drugs.

NSAIDs mechanism of action: it can inhibit activity of COX, thereby reducing synthesis of prostaglandin PGs, and achieving anti-inflammatory and analgesic effects. The COX enzyme has two isoenzymes, namely basic expression COX-1, which has the main function of maintaining the normal physiological functions of organs such as stomach, intestine, kidney and the like; COX-2, expressed by induction, can be found and act locally in the brain and inflammation. The anti-inflammatory mechanism of action of NSAIDs is thought to be inhibition of COX-2, while adverse effects occur due to inhibition of COX-1.

Common nonspecific NSAIDs such as ibuprofen (ibuprofen), naproxen (naproxen), diclofenac sodium and the like have long and narrow structures and no selectivity, and can be combined with the active sites of COX-1 and COX-2 simultaneously, so that the COX-1 and the COX-2 can be inhibited. Therefore, not only has the functions of diminishing inflammation and easing pain, but also has adverse reactions such as gastrointestinal tract, blood coagulation function, renal function and the like.

The major problem with NSAIDs treatment is adverse effects. With the increasing age, the adverse reactions are more remarkable, including gastrointestinal tract, kidney, cardiovascular system and central nervous system.

Progress in the study of (tetra) viral diseases and their treatment

Statistically, 60% to 65% of epidemic diseases are caused by viral infections. The seventh virology report, presented by the world virology congress summoned in paris, france, 8 months 2002, indicated that there were 29 families, 7 subfamilies, 53 residences, and 1200 viruses that were pathogenic to humans. The incidence rate is high, 6 viral diseases with great harmfulness are available, such as acute and chronic hepatitis caused by hepatitis virus; cytomegalovirus (CMV) retinitis, interstitial pneumonia, herpes encephalitis, keratitis, genital herpes, herpes zoster, and herpes labialis caused by 8 kinds of human herpesviruses; bronchitis, pneumonia, measles, mumps, poliomyelitis caused by respiratory virus infection; acute gastroenteritis and infantile diarrhea caused by enterovirus, AIDS caused by Human Immunodeficiency Virus (HIV), etc.

Although humans are constantly searching for effective weapons against viruses since the first day of virus recognition, treatment of viral diseases is still lacking in highly specific drugs. The following drugs are commonly used clinically: antiviral agents that inhibit viral replication; ② an immunity regulating medicine for enhancing the immunity function of the organism; ③ cough-relieving, pain-easing, fever-abating and inflammation-diminishing therapeutic drugs aiming at clinical symptoms; anti-infective medicine for preventing secondary infection; vaccine for preventing virus infection; sixth, disinfectant to block virus transmission.

(V) analgesic drug research progress

Pain is a common sensation among humans and widely different among individuals, and is an alarm signal that the body and internal organs are damaged. Pain is much more difficult to study than other sensations because it is often interlaced with autonomic nervous activity, psychological and emotional responses, and has great variability. The etiology and pathology of pain, including migraine, rheumatoid arthritis, osteoarthritis, back pain, cancer pain, and neuropathic pain, has been poorly understood for a long time, and thus, the treatment of pain, particularly neuropathic pain, caused by most diseases remains difficult. Although the efforts have been made for more than 30 years, the problem has not been solved satisfactorily, and the fundamental reason is that the understanding of etiology and pharmacology of pain is slow. The most commonly used analgesics are non-steroidal anti-inflammatory drugs (abbreviated as NSAIDs) and opioids, and there are 18 kinds of NSAIDs in common use, and up to 100 kinds of NSAIDs are under study. Although they have good clinical effects, adverse reactions such as gastrointestinal reactions of NSAIDs, gastric ulcers, gastrorrhagia, and allergies are daunting. Opioids are restricted by law due to their addictive nature. In particular, the treatment difficulty of the neuropathic pain caused by nerve injury or lesion caused by nociceptive pain (nociceptive pain), complex regional pain syndrome (CRPS for short) and trauma, inflammation or other diseases is higher, and the powerful narcotic analgesic is difficult to be effective. It is feared that the understanding of etiology and pharmacology of chronic pain has been developed in recent years, which makes the concept of pathology, physiology and therapeutics of chronic pain changed fundamentally, and breaks away from the old frame of traditional NSAIDs and opioids, and develops a batch of novel analgesics. But most are in development or clinical trials. In addition, in recent years, chinese scholars have made a lot of work in developing therapeutic drugs for neuropathic pain with the target of N-type voltage sensitive calcium ion channels, nerve growth factor receptors, adenosine kinase, substance P receptors, and cannabinoid receptors, and the search for analgesic agents for treating neuropathic pain has become a leading issue of clinical and basic research and therapeutic drug research.

Research progress of pharmacological activity of (hexa) phenolic acid compound

1. The research results of seven water-soluble components of salvia miltiorrhiza for resisting platelet aggregation and preventing thrombosis for intravenous injection to rats on platelet aggregation of rats show that the salvianolic acid A and the salvianolic acid B have stronger inhibition effects on collagen-induced platelet aggregation, the IC50 of the salvianolic acid A and the IC50 of the salvianolic acid B are 0.32mmol/L and 0.26mmol/L respectively, and the salvianolic acid A is effective and the salvianolic acid B is ineffective on ADP-induced aggregation. Rosmarinic acid has mild effect in preventing thrombosis, and is one of effective components of Saviae Miltiorrhizae radix with antithrombotic effect. Rosmarinic acid (iv)50mg/kg, 100mg/kg inhibited venous thrombosis in rats at 41% and 55.8% (P < 0.05).

2. The change conditions of the caliber, the blood flow speed and the flow of mouse mesenteric arteriole vessels under different conditions are observed to show that the local administration of the salvianolic acid A and the salvianolic acid B has obvious improvement effect on the mouse mesenteric microcirculation disturbance caused by epinephrine, and the intravenous injection of the salvianolic acid A also has the same effect.

3. The lipid peroxidation resistant salvianolic acid A, salvianolic acid B, salvianolic acid C and rosmarinic acid have stronger inhibition effect on the generation of mouse liver homogenate malondialdehyde (MDA for short) than vitamin E, and the inhibition intensity is that of salvianolic acid A>Salvianolic acid C>Salvianolic acid B>The four components of the rosmarinic acid have 10-100 times stronger scavenging effect on hydroxyl free radicals than mannitol. For mice suffering from acute alcoholism caused by oral administration of 15ml/kg of 50% ethanol, the MDA content in the liver of the mice is remarkably reduced after the mice are treated by rosmarinic acid (100mg/kg multiplied by 3, po). Salvianolic acid A and salvianolic acidThe acid B and rosmarinic acid pair is composed of vitamin C-Nicotinamide Adenine Dinucleotide Phosphate (NADPH) or Fe²⁺Cysteine-induced lipid peroxidation of rat brain, liver and kidney microsomes has strong inhibition effect, and the effect is sequentially salvianolic acid A, salvianolic acid B and rosmarinic acid, which is 100-1000 times stronger than that of antioxidant vitamin E. Salvianolic acid A can inhibit Fe²⁺Cysteine-induced mitochondrial lipid peroxidation and ATPase activity loss in rat heart and liver, inhibition of lipid peroxidation-induced mitochondrial swelling and decreased liver mitochondrial membrane fluidity.

4. Prevention of doxorubicin cardiotoxicity is caused by free radical production in the heart and damage to myocardial mitochondria. The salvianolic acid A can eliminate OH generated by adriamycin, has a prevention effect on cardiotoxicity caused by adriamycin, and has no influence on antitumor effect of adriamycin.

5. The salvianolic acid A has effect in inhibiting pig stomach H⁺-K⁺Effect of ATP-and PNPP-enzymes, IC₅₀Are respectively 5.2 multiplied by 10^-7And 1.7X 10^-6mol/L. Salvianolic acid A (25mg/kg, ip) has strong inhibitory effect on gastric acid secretion of pylorus-ligated rat, and the same dosage of Salvianolic acid A can reduce gastric injury caused by water immersion or forced stress. These results indicate that salvianolic acid A inhibits gastric H⁺-K⁺ATP enzyme has gastric acid secretion inhibiting and antiulcer effects.

6. The magnesium salt of the salvianolic acid B for relieving uremia can reduce the concentration of urea nitrogen, creatinine, guanidine methyl amidine and guanidyl succinic acid in the blood of mice with uremia caused by taking adenine, thereby obviously relieving the uremia symptoms of the mice.

7. Inhibiting adenylate cyclase rosmarinic acid, alkannic acid and methyl ester thereof has strong inhibiting effect on basic adenylate cyclase of rat brain and adenylate cyclase activated by 5umol/L diterpene derivative.

8. Other compounds such as ginsenoside and some phenolic acid are main active ingredients of ginseng for resisting aging; tea polyphenols are a general term for a variety of phenolic derivatives, accounting for about 25%, and include catechins, anthocyanidins, flavones, flavonols, and phenolic acid quats. They contain polyhydroxy, have the property of phenol, and have remarkable pharmacological actions, such as reducing blood sugar and blood fat, and preventing atherosclerosis; can be combined with proteins of bacteria such as Escherichia coli and streptococcus and pathogenic bacteria such as typhoid fever and cholera in human body to precipitate, inhibit growth of bacteria, and promote multiple bacteria to lose activity; can also precipitate toxic substances such as nicotine, dehydrated morphine, cinchona alkaloid and the like in human bodies and heavy metals such as lead, cobalt, silver, copper and the like harmful to the human bodies, and reduce or eliminate the harmfulness.

According to the literature search and the like, no report on tracking and screening the active site or the active ingredient of the single cocklebur fruit exists so far, and no report on the application of the cocklebur fruit active site extract is found.

Disclosure of Invention

The technical problem to be solved by the invention is to disclose a new application of a total extract of traditional Chinese medicine fructus xanthil, so as to overcome the defects in the prior art.

That is, the invention aims to clarify the active site and active ingredient of the traditional Chinese medicine fructus xanthil and the preparation method thereof, and further use the total extract of the fructus xanthil for preparing anti-inflammatory reaction products; the invention relates to a traditional Chinese medicine cocklebur fruit total glycoside extract and a preparation method thereof, and application of the total glycoside extract and a composition thereof, such as a pharmaceutical composition, in preparation of anti-inflammatory reaction products.

The anti-inflammatory reaction product is a product for resisting inflammation and related diseases, a product for resisting virus and related diseases, a product for easing pain and related diseases, a product for diagnosing, detecting, treating or researching acute and chronic rhinitis and related diseases, or a product for diagnosing, detecting, treating or researching nasosinusitis and related diseases; wherein the anti-inflammatory reaction product is one or more selected from anti-inflammatory reaction drug, anti-inflammatory reaction reagent, anti-inflammatory reaction food or anti-inflammatory reaction beverage, preferably anti-inflammatory reaction drug.

Technical idea

The independent development of innovative drugs is an urgent task in China at present. Chinese traditional medicine has a long history, and abundant experiences are accumulated in the aspect of preventing and treating diseases by using Chinese herbal medicines, so that the search of effective active ingredients from the Chinese traditional medicine is an effective way and is the advantage of developing innovative Chinese medicines.

The inventor searches books such as 'new edition Chinese patent medicine', 'handbook of clinical commonly used prescription' and 'Chinese commonly used Chinese patent medicine' and the like, and carries out investigation statistical analysis on hospital medication and commercially available drugs of patients in the department of five sense organs, and finds that among 30 commonly used Chinese medicine prescriptions for treating rhinitis and nasosinusitis, 23 xanthium seeds are contained, account for 76.7%, and are all used as monarch drugs or ministerial drugs and are the key drugs for the department of rhinology. It is therefore assumed that substances or substance groups with anti-inflammatory, antibacterial and antiviral activity are very likely to be present in xanthium sibiricum. However, due to the fact that the research on chemical components is not deep, the pharmacological screening is insufficient, the active site and the active components are not clear, and no feasible quality standard exists, and the further development and utilization of the cocklebur fruit are limited.

Therefore, the invention carries out systematic chemical component research on the total extract of the single cocklebur fruit medicinal material, and screens and proves the activity and the application of the total extract.

Chemical component research proves that the main components in the active site of the cocklebur fruit are diterpene glycoside compounds and phenolic acid compounds. According to literature search, the inventors find that most of phenolic acid compounds have various remarkable pharmacological activities, such as famous salvianolic acid, ginkgolic acid and the like (see background technology), so that the inventors presume that the anti-inflammatory and analgesic and antiviral activities of the cocklebur fruit and the clinical efficacy of treating various nasal diseases are mainly exerted through the efficacy of the active site cocklebur fruit total glycoside, and research results also prove and prove that the cocklebur fruit total glycoside extract has remarkable pharmacological activity.

Extract of total glycosides of Xanthium strumarium

1. Composition of cocklebur fruit total glycoside extract

Through the experimental research of systematic chemical component separation, purification, structure identification and the like on the cocklebur fruit medicinal material, the inventor finds that the cocklebur fruit mainly contains a large amount of volatile oil, sesquiterpene lactone and water-soluble glycoside compounds.

The components in the fructus Xanthii total glycoside extract mainly include diterpene glycoside compounds and phenolic acid compounds; the two compounds are main effective components in the fructus xanthil total glycoside extract and also comprise other components.

Wherein, the content range of diterpene glycoside compounds in the total glycoside extract is 0.5-99% (W/W, weight percentage), preferably 5.0-95.0%;

wherein the content of phenolic acid compounds in the total glycoside extract is 0.5-99% (W/W), preferably 5.0-95.0%.

Wherein the content of other components in the total glycoside extract is below 0.5%, and the other components include one or more of tannin, monosaccharide or polysaccharide, xanthones (xanthones), carotenoid, etc.

The existing research shows that other components at least have no negative influence on the drug effect, the use of the drug and the like, and the cost for separating the components is high, so that further purification is not necessary at present.

2. The general chemical structure formula of diterpene glycoside compounds in the cocklebur fruit is as follows:

wherein,R₁is one of hydrogen, methyl or carboxyl; r₂、R₃Is one of hydrogen, sulfonic acid group, acetyl group, propionyl group or dimethylpropionyl group.

Table 1 shows the structural formulas of some diterpene glycosides, and R₁、R₂、R₃Correspondence of groups:

when R is_I＝COOH，R₂＝R₃When H, represents diterpene glycoside compound a;

when R is₁＝COOH，R₂＝H，R₃＝COCH₂CH(CH₃)₂When the compound is a diterpene glycoside compound B;

when R is₁＝COOH，R₂＝SO₃ ^-，R₃＝COCH₂CH(CH₃)₂When the formula (I) is shown in the specification, the formula (I) represents a diterpenoid glycoside compound C;

when R is₁＝H，R₂＝H，R₃＝COCH₂CH(CH₃)₂When the formula (I) is shown in the specification, the formula (I) represents a diterpenoid glycoside compound D;

when R is₁＝H，R₂＝SO₃ ^-，R₃＝COCH₂CH(CH₃)₂When the formula (I) is used, the formula (I) represents a diterpene glycoside compound E.

TABLE 1 chemical structural formula of diterpene glycoside compounds in fructus Xanthii

	R<sub>1</sub>	R<sub>2</sub>	R<sub>3</sub>
	R<sub>1</sub>	R<sub>2</sub>	R<sub>3</sub>	A	COOH	H	H
B	COOH	H	-COCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>	A	COOH	H	H
B	COOH	H	-COCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>	C	COOH	SO<sub>3</sub><sup>-</sup>	-COCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
D	H	H	-COCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>	C	COOH	SO<sub>3</sub><sup>-</sup>	-COCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
D	H	H	-COCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>	E	H	SO<sub>3</sub><sup>-</sup>	-COCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>

3. The general chemical structure of the phenolic acid compound in the cocklebur fruit is as follows:

wherein R is₁Is one of hydrogen or alkali metal elements; r₂、R₃And R₄Is one of hydrogen or caffeoyl.

Caffeoyl, Caffeoyl, has the following structure:

TABLE 2 chemical structural formulas of several phenolic acid compounds in fructus Xanthii

	R<sub>1</sub>	R<sub>2</sub>	R<sub>3</sub>	R<sub>4</sub>
	R<sub>1</sub>	R<sub>2</sub>	R<sub>3</sub>	R<sub>4</sub>	A	H	Caffeoyl	Caffeoyl	Caffeoyl
B	H	Caffeoyl	H	Caffeoyl	A	H	Caffeoyl	Caffeoyl	Caffeoyl
B	H	Caffeoyl	H	Caffeoyl	C	H	H	Caffeoyl	Caffeoyl
D	K	H	Caffeoyl	H	C	H	H	Caffeoyl	Caffeoyl

Table 2 shows the structural formulas of some phenolic acid compounds₁、R₂、R₃The corresponding relation of the groups.

When R is₁＝H，R₂＝R₃＝R₄When Caffeoyl, it represents a phenolic acid compound A;

when R is₁＝R₃＝H，R₂＝R₄When Caffeoyl, it represents a phenolic acid compound B;

when R is₁＝R₂＝H，R₃＝R₄When Caffeoyl, it represents a phenolic acid compound C;

when R is₁＝K，R₂＝R₄＝H，R₃When Caffeoyl is substituted, phenolic acids are representedCompound D.

Preparation method of total glycoside extract of fructus Xanthii

The preparation method of the cocklebur fruit total glycoside extract comprises the following steps;

(1) extraction: taking a plurality of cocklebur fruit raw materials, and extracting to obtain an extracting solution, namely a total cocklebur fruit extract;

(2) separation and purification: separating and purifying the extracting solution to obtain the cocklebur fruit total glycoside extract.

The extraction method includes all methods known in the art such as solvent extraction.

The extraction method of the solvent extraction method described above is a conventional extraction method in the art, that is, includes one or more of a commonly used ultrasonic extraction method, an immersion extraction method, a percolation extraction method, a decoction extraction method, a heating reflux extraction method, a continuous reflux extraction method, or the like; the number of extractions may be one or more. Meanwhile, the extraction efficiency can be influenced by the factors such as the crushing degree of the raw materials, the extraction time, the extraction temperature, the equipment conditions and the like, and the factors need to be considered. All technical data including technical details of various extraction methods can be found in relevant textbooks, relevant technical documents and the like.

The extraction solvent used in the solvent extraction method described above is a conventional extraction solvent in the art, that is, one or more of three types of extraction solvents including common aqueous reagents, hydrophilic organic solvents, or lipophilic organic solvents;

the water reagent is one of water, acid water, alkaline water and the like;

the hydrophilic organic solvent is generally called water-miscible organic solvent, and includes one or more of ethanol, ethanol aqueous solution, methanol and the like, preferably one or more of ethanol or ethanol aqueous solutions with different concentrations and the like, and further preferably ethanol aqueous solutions with different concentrations;

the lipophilic organic solvent is generally called water-immiscible organic solvent, and comprises one or more of petroleum ether, chloroform, diethyl ether, ethyl acetate, dichloromethane or dichloroethane, preferably one or more of petroleum ether, chloroform, diethyl ether, ethyl acetate, dichloromethane or dichloroethane, and more preferably one or more of chloroform, diethyl ether, ethyl acetate or dichloromethane.

The total extract of fructus Xanthii obtained by the above extraction method needs further separation and purification.

The separation and purification method is all available methods known in the art, and comprises one or more of a solvent separation method, a solvent extraction method (comprising one or more of an extraction method, a countercurrent continuous extraction method, a countercurrent distribution method or a droplet countercurrent distribution method, and the like), a macroporous adsorption resin method, a precipitation method, a salting-out method, a column chromatography method or a crystallization and recrystallization method, a fractional crystallization method, and the like; these methods are well known in the art, and all technical data including technical details can be easily found from relevant textbooks and relevant technical documents when further confirmation is needed.

In order to stabilize the product properties, facilitate the use and the storage, and improve the bioavailability of the product, an optimization step can be added, namely the step three:

(3) and (5) drying.

The drying method is a method known in the art and can be used, and includes one or more of an atmospheric drying method such as oven drying, a reduced pressure drying method, a vacuum drying method, a spray drying method, a freeze drying method, a far infrared heating drying method, or a microwave drying method.

In the specific implementation, necessary measures such as adopting a proper method and selecting proper conditions according to the existing funds, technologies and related requirements are needed to achieve the expected targets. These methods are well known in the art, and all technical data including technical details can be easily found from relevant teaching materials and relevant technical documents when further confirmation is needed.

Application of total glycoside extract of (IV) cocklebur fruit

1. Overview

A further object of the present invention is to provide a novel use of the extract of the total glycosides of Xanthium strumarium in the preparation of anti-inflammatory products, including one or more of drugs, reagents, foods or beverages, preferably drugs.

Pharmacological activity screening proves that the cocklebur fruit total glycoside extract obtained by adopting a n-butyl alcohol extraction part or a macroporous adsorption resin 5-75% ethanol elution part is an active part for resisting inflammation, easing pain, resisting virus or diagnosing, detecting, treating and researching rhinopathy and related diseases. The completed acute toxicity experiment proves that the maximum tolerance of the mouse gavage administration to the active part exceeds 2.0g/kg, which is equivalent to 440 times of the clinical recommended dosage, shows that the effective part is safe and reliable, and solves the problems of complex components, low content of effective components and toxic components in the traditional Chinese medicine compound.

The inventor finds that the cocklebur fruit total glycoside extract has anti-inflammatory effect and analgesic effect, can obviously reduce inflammation swelling degree and inhibit capillary permeability, and has antiviral effect.

Meanwhile, the inventor finds that the cocklebur fruit total glycoside extract can relieve the nasal mucosal inflammation cell infiltration caused by rhinitis and nasosinusitis, eliminate nasal mucosa edema, inhibit the proliferation of connective tissue under mucosa so as to improve the nasal mucosa swelling and ventilation function, reduce nasal secretions so as to eliminate inflammation, and the effect is better than that of the clinical commonly used medicine nasal sinusitis oral liquid and the like.

2. Methods of use and requirements for cocklebur fruit total glycoside extracts and compositions thereof

The cocklebur fruit total glycoside extract can be used alone or combined with other active components, and comprises products for diagnosing, detecting, treating or researching related diseases, including medicaments, reagents, foods or beverages, and the like, in particular medicaments.

In a specific use aspect, the cocklebur fruit total glycoside extract can be used alone, and can also be used together with other various chemical substances. Whether the chemical substances have biological activity or disease treatment functions, including auxiliary functions such as synergistic amplification, antagonism or alleviation of side effects of the cocklebur fruit total glycoside extract, and the like, the chemical substances comprise one or more of pharmaceutically acceptable carriers, foods, natural products, chemically synthesized drugs or human drugs; preferably comprises one or more of pharmaceutically acceptable carriers or food, etc.; further preferably a pharmaceutically acceptable carrier.

As used herein, "pharmaceutically acceptable carrier" includes any and all physiologically acceptable solvents, dispersion media, integuments, antibacterial and antifungal agents, isotonic or absorption delaying agents, and the like. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol or ethanol, and the like, and combinations thereof. In many cases, it will be desirable to include isotonic agents, for example, one or more of sugars, polyalcohols such as mannitol, sorbitol, sodium chloride and the like in the composition. The pharmaceutically acceptable carrier may also contain minor amounts of auxiliary substances, such as one or more of wetting or emulsifying agents, preservatives or buffers, and the like, which enhance the effective period or effectiveness of the cocklebur fruit total glycoside extract.

In a specific classification, the pharmaceutically acceptable carrier refers to a conventional pharmaceutical carrier in the pharmaceutical field, and includes excipients, such as one or more of starch, water and the like; one or more of a lubricant, such as glycerin or magnesium stearate, and the like; disintegrants, such as microcrystalline cellulose and the like; fillers, such as one or more of starch or lactose; a binder such as one or more of pregelatinized starch, dextrin, cellulose derivatives, alginate, gelatin, or polyvinylpyrrolidone, etc.; osmotic pressure regulators, such as one or more of glucose, sucrose, sorbitol, or mannitol; diluents such as water and the like; disintegrating agents, such as one or more of agar, calcium carbonate or sodium bicarbonate; absorption accelerators such as quaternary ammonium compounds and the like; surfactants such as cetyl alcohol and the like; an adsorption carrier, such as one or more of kaolin, bentonite, etc.; lubricants, such as one or more of talc, calcium stearate, magnesium stearate, or polyethylene glycol; in addition, other adjuvants such as one or more of flavoring agents or sweeteners may also be added to the composition.

For example, the injection preparation can be prepared by dissolving, suspending or emulsifying the active ingredient cocklebur fruit total glycoside extract in a suitable aqueous solvent (e.g., one or more of distilled water, physiological saline, or a solution of green) or oily solvent (e.g., one or more of vegetable oil such as olive oil, sesame oil, cottonseed oil, corn oil, or propylene glycol), wherein the solvent may contain a dispersing agent (e.g., one or more of polysorbate 80, polyoxyethylene hardened castor oil 60, polyethylene glycol, benzyl alcohol, chlorobutanol, or phenol), an osmotic pressure regulator (e.g., one or more of sodium chloride, glycerol, D9-mannose, D-sorbitol, or glucose, etc.). In this case, additives such as solubilizing agents (e.g., one or more of sodium salicylate, sodium acetate, or the like), stabilizers (e.g., human serum albumin, or the like), analgesics (e.g., benzyl alcohol, or the like), and the like may be added, if necessary.

The cocklebur fruit total glycoside extract described in the present invention may also be used in combination in the form of a composition, in particular a composition for the treatment of animals, especially mammals including humans or other animals, with other chemical substances such as drugs, or similar compositions. The mammal comprises one or more of human, mouse, rat, sheep, monkey, cow, pig, horse, rabbit, dog, chimpanzee, baboon, marmoset, macaque or rhesus monkey. For example, the cocklebur fruit total glycoside extract of the present invention may be incorporated into a pharmaceutical composition suitable for administration to a subject. Generally, the pharmaceutical composition comprises the cocklebur fruit total glycoside extract and a pharmaceutically acceptable carrier.

Compositions of cocklebur fruit total glycoside extract, particularly pharmaceutical compositions, may be in various forms including one or more of dosage forms such as liquid, semi-solid and solid; the medicine composition comprises a therapeutically effective amount of cocklebur fruit total glycoside extract as an active ingredient and one or more pharmaceutically acceptable carriers.

The pharmaceutical composition of the cocklebur fruit total glycoside extract can be prepared into various dosage forms by adopting the conventional production method known in the field, for example, the active ingredient is mixed with one or more carriers and then is prepared into the required dosage form. The dosage form comprises one or more of tablets, capsules, granules, suspensions, emulsions, solutions, syrups or injections and the like, and one or more administration routes of oral administration or injection (including one or more of intravenous injection, intravenous drip, intramuscular injection or subcutaneous injection and the like), mucosal dialysis and the like are adopted for anti-inflammatory, antiviral and analgesic and diagnosis, detection, treatment or scientific research of acute and chronic rhinitis, nasosinusitis and related diseases.

The pharmaceutical composition preferably contains 0.5 to 99 weight percent of the active ingredient cocklebur fruit total glycoside extract, more preferably contains 1 to 95 weight percent of the active ingredient cocklebur fruit total glycoside extract, and most preferably contains 5 to 90 weight percent of the active ingredient cocklebur fruit total glycoside extract.

Pharmaceutical compositions of cocklebur fruit total glycoside extracts generally must be sterile and stable under the conditions of manufacture and storage. The compositions may be formulated as solutions, microemulsions, dispersions, liposomes or other ordered structures suitable for high drug concentrations. Sterile injectable solutions are prepared by incorporating the desired amount of the cocklebur fruit total glycoside extract in the appropriate solvent with one or a combination of the desired ingredients enumerated above, followed by sterile filtration. Generally, the dispersion is prepared by adding the cocklebur fruit total glycoside extract to a sterile vehicle containing a basic dispersion medium and the required other ingredients described above. In the case of sterile powders for the preparation of sterile injectable solutions, the recommended methods of preparation are vacuum drying and freeze drying. Proper fluidity of the solution can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants. Prolonged absorption of the injectable compositions can be achieved by including in the composition an agent which delays absorption, for example, monostearate salts or gelatin.

When the fructus xanthil total glycoside extract is applied to a patient, the dosage of the fructus xanthil total glycoside extract is 5-20 mg/kg d, and the dosage or the dosage is usually determined according to the age and the weight of the patient or a user and the physical condition or the condition of symptoms of the patient.

The cocklebur fruit total glycoside extract and the medicinal composition thereof can comprise a 'treatment effective amount' or a 'prevention effective amount' of the cocklebur fruit total glycoside extract. A "therapeutically effective amount" is an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic effect. The therapeutically effective amount of the cocklebur fruit total glycoside extract may vary depending on factors such as the condition, age, sex and weight of the individual and the ability of the cocklebur fruit total glycoside extract to elicit a desired response in the individual. A therapeutically effective amount also refers to an amount of the total glycoside extract of xanthium sibiricum that has a beneficial therapeutic effect over any toxic or detrimental effects thereof. A "prophylactically effective amount" is an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic effect. Because prophylactic doses are used in subjects before or at an early stage of the disease, the prophylactically effective amount is generally less than the therapeutically effective amount. The typical non-limiting range of the effective amount of the cocklebur fruit total glycoside extract for treatment or prevention is 5-20 mg/kg, and the more preferable range is 5-10 mg/kg. It is to be noted that the dosage value will vary depending on the type and severity of the disease to be alleviated, that is, when administered to a patient, the dosage or amount of the total glycoside extract of Xanthium sibiricum of the present invention will generally be determined according to the age and weight of the patient or user and the physical condition or condition of the patient's symptoms. In addition, it is to be understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the dosage ranges set forth herein are exemplary only and do not limit the scope or practice of the claimed compositions.

That is, the dose or amount per time and/or day of the cocklebur fruit total glycoside extract of the present invention is required to be varied depending on the subject to be treated, the route of administration, the disease and condition to be treated, and the like. For example, the cocklebur fruit total glycoside extract is intravenously administered to a mammal, especially an adult (such as 60kg body weight), and the single dose of the cocklebur fruit total glycoside extract is about 5-10 mg, preferably about 10mg, and preferably 1-3 times daily administration. Dosage units may be adjusted to provide the optimum desired response (e.g., therapeutic or prophylactic response). For example, a bolus dose may be given, several aliquots may be given over a period of time or the dose may be proportionally reduced or increased depending on the exigencies of the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form, as used herein, refers to physically discrete units of unit dosage suitable for the mammalian subject to be treated; each unit containing a predetermined amount of the active cocklebur fruit total glycoside extract calculated to produce the desired therapeutic effect in association with the desired pharmaceutical carrier. The specification for the dosage unit forms of the present invention is determined by and directly depends on the following (a) unique characteristics of the cocklebur fruit total glycoside extract and the specific therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the art of compounding such a total glycoside extract for the treatment of sensitive cocklebur fruit in an individual.

3. Pharmaceutical dosage form and administration route of fructus Xanthii total glycoside extract and composition thereof

The cocklebur fruit total glycoside extract and the composition thereof are used for preparing anti-inflammatory reaction products, wherein the products prepared according to the requirements of the technical field of beverages and foods can resist inflammation, virus and pain, and diagnose, detect, treat or research acute and chronic rhinitis, nasosinusitis and related diseases; the product prepared according to the requirements of the medical technical field can be used for treating or protecting the health of patients, can be directly used for preparing medicaments for treating or protecting the health independently, and can also be mixed or combined with a plurality of chemical substances to be directly or indirectly used for preparing the medicaments for treating or protecting the health. The chemistry described herein is the same as described above in this section.

In the invention, the required materials comprise the raw materials of the invention, the chemical substances used in coordination with the raw materials and the like, and food-grade or medicinal-grade materials are adopted according to actual conditions and requirements.

The cocklebur fruit total glycoside extract and compositions thereof according to the present invention may be administered by various methods known in the art, although the recommended route/mode of administration in many therapeutic applications is by spray or oral administration. However, the skilled artisan will appreciate that the route/mode of administration will vary depending on the desired result. In certain implementations, the active compound can be formulated, for example, as an empty release formulation, including one or more of a graft delivery system, a transdermal patch delivery system, or a microencapsulated delivery system, along with a carrier that protects the compound from rapid release. In addition, biodegradable, biocompatible polymers may also be used, such as one or more of ethyl vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, or polylactic acid, among others. Many methods of preparing such formulations are patented or generally known to those skilled in the art (see, e.g., Sustained and Controlled Release Drug Delivery Systems, J.R. Robinson, eds., Marcel Dekker, Inc., New York, 1978).

The cocklebur fruit total glycoside extract and the composition thereof are generally applied to a patient needing the treatment by one or more of oral administration, nasal inhalation, rectal administration or parenteral administration and the like.

For oral administration, it may be formulated into one or more conventional solid preparations such as tablets, powders, granules or capsules. In practice, the cocklebur fruit total glycoside extract of the present invention may be administered orally, for example, with an inert diluent or an assimilable edible carrier. The cocklebur fruit total glycoside extract (and other ingredients, if desired) may also be encapsulated in hard or soft shell gelatin capsules, compressed into tablets or added directly to the subject's diet. For oral therapeutic administration, the cocklebur fruit total glycoside extract may be added with excipients and used in the form of one or more of an edible tablet, buccal tablet, tincture, capsule, suspension, syrup, wafer, or the like.

In order to administer the cocklebur fruit total glycoside extract of the present invention in addition to parenteral administration, it may be necessary to coat or administer the cocklebur fruit total glycoside extract with a material that prevents its inactivation. Supplementary active compounds may also be added to the composition. In particular implementations, the cocklebur fruit total glycoside extract of the present invention is co-formulated and/or co-administered with one or more other therapeutic agents that may be used to treat a disease. Such a combination may advantageously utilize lower doses of the administered therapeutic agent, thus avoiding possible toxicity or complications associated with various monotherapies.

Making into liquid preparation such as aqueous solution, oil suspension or other liquid preparation, such as syrup or elixir; for parenteral administration, it may be formulated into one or more of solution for injection, aqueous solution, oily suspension, etc.

Among the above-mentioned use forms, preferred forms are one or more of tablets, coated tablets, capsules, suppositories, nasal sprays, nasal drops, injections and the like, further preferred are one or more of nasal sprays, nasal drops, tablets and the like, and particularly preferred are nasal sprays.

In addition, the raw materials of the cocklebur fruit total glycoside extract can be directly used for preparing anti-inflammatory reaction products independently under certain conditions, and can also be mixed or combined with a plurality of chemical substances to be directly or indirectly used for preparing the anti-inflammatory reaction products in the form of compositions. The chemistry described herein is the same as described above in this section.

For example, the powder of the raw materials used for the extract of the cocklebur fruit total glycosides is used for preparing various dosage forms of anti-inflammatory reaction products, especially drugs, or the powder of the raw materials used for the extract of the cocklebur fruit total glycosides and related auxiliary materials are used for preparing various dosage forms of anti-inflammatory reaction products, especially drugs, or the powder of the raw materials used for the extract of the cocklebur fruit total glycosides and related prepared anti-inflammatory reaction products, such as drugs, are used together for preparing various dosage forms of anti-inflammatory reaction products, such as drugs, or the powder of the raw materials used for the extract of the cocklebur fruit total glycosides and related auxiliary materials are used together for preparing various dosage forms of anti-inflammatory reaction products, such as drugs, such as one or more of tablets, capsules, granules or suspensions, and the like, preferably capsules.

The other method is that the powder of the medicinal raw material used by the cocklebur fruit total glycoside extract and related anti-inflammatory reaction products such as medicines are filled into capsules; the fourth method is to directly compress the powder of the raw materials used by the cocklebur fruit total glycoside extract and related auxiliary materials into tablets or granules according to a conventional method, the fifth method is to directly compress the powder of the raw materials used by the cocklebur fruit total glycoside extract, related anti-inflammatory reaction products such as medicines and related auxiliary materials into tablets or granules according to a conventional method, and the sixth method is to directly compress the powder of the raw materials used by the cocklebur fruit total glycoside extract, related auxiliary medicines and related auxiliary materials into tablets or granules according to a conventional method.

Besides the six basic methods, other forms of the medicinal raw materials used by the cocklebur fruit total glycoside extract or products such as medicines containing the medicinal raw materials used by the cocklebur fruit total glycoside extract in various dosage forms can be prepared after the medicinal raw materials used by the cocklebur fruit total glycoside extract are processed by methods known in the art. However, it should be noted that, when the raw materials used in the cocklebur fruit total glycoside extract are directly used, the dosage of the raw materials used in the cocklebur fruit total glycoside extract to be used should be converted according to the dosage requirement of the cocklebur fruit total glycoside extract to be used.

In conclusion, the cocklebur fruit total glycoside extract and the composition thereof can be used for preparing anti-inflammatory reaction products, preferably medicaments and foods, and further preferably medicaments.

(V) technical speciality

The invention researches the cocklebur fruit total glycoside extract with pertinence, the raw materials are safe to use, give consideration to each other, have multiple purposes, exert the function to the maximum extent, and have extremely wide application range, so the invention is easy to popularize and apply, and can generate great social benefit and economic benefit in a short time.

That is, the research result of the invention can be further developed into the products with clinical application prospect for preventing and treating rhinitis, nasosinusitis, anti-inflammation, analgesia and antivirus, in particular to new drugs. The product has the advantages of definite curative effect, low effective dose and no toxic or side effect, and has obvious advantages compared with similar products at home and abroad in China. Acute and chronic nasosinusitis are common and frequently encountered diseases in clinic, and with the increasing severity of global air pollution and the deterioration of the environment, the incidence rate of respiratory diseases such as nasosinusitis and the like is expected to rise continuously. Therefore, the invention can lay a good foundation for the subsequent new drug research and further generate greater economic and social benefits.

Through long-term pharmacological tests, the cocklebur fruit total glycoside extract has the activities of resisting inflammation, resisting virus, easing pain and treating acute and chronic rhinitis, nasosinusitis and related diseases, the cocklebur fruit total glycoside extract is directly used, the metabolism of gastrointestinal tract bacteria is not needed, the effect is more direct, the difference of drug effect caused by the difference of metabolites due to the difference of gastrointestinal tract flora activity among individuals is avoided, the problem that the administration dosage of the traditional Chinese medicine is difficult to control is solved, and the bioavailability is improved.

The cocklebur fruit total glycoside extract has strong pharmacological action, rich raw material sources, simple preparation process and high yield.

The cocklebur fruit total glycoside extract is also an active form in gastrointestinal tract biotransformation, and the direct use of the cocklebur fruit total glycoside extract has high bioavailability and can accurately control the dosage. The fructus Xanthii total glycoside extract has stable properties, and the preparation prepared from the fructus Xanthii total glycoside extract has stable quality.

In addition, the fructus xanthil total glycoside extract has stable chemical properties, and has obvious effects of resisting inflammation, viruses and pain and treating acute and chronic rhinitis, nasosinusitis and related diseases, so the fructus xanthil total glycoside extract is more suitable for the industrial production of medicaments for resisting inflammation, viruses and pain and treating acute and chronic rhinitis, nasosinusitis and related diseases.

In conclusion, the invention actively adapts to the working needs and the needs of humanized services in the fields of modern medical treatment, food, beverage and scientific research, is a safe raw material for preparing anti-inflammatory reaction products, can directly or indirectly resist inflammation, virus and pain, and can diagnose, detect, treat or research acute and chronic rhinitis, nasosinusitis and related diseases.

Detailed Description

The invention researches the application of the cocklebur fruit total glycoside extract, provides a raw material which can be used for preparing anti-inflammatory reaction products such as medicines and the like or anti-inflammatory reaction products, particularly medicines, and is convenient for safe use in the medical industry and related industries such as the fields of foods, beverages and the like, including diagnosis, detection, treatment or scientific research of diseases and the like.

The following details the specific operation method for preparing the cocklebur fruit total glycoside extract by taking the solvent extraction method as an example.

(1) Extraction: taking a plurality of cocklebur fruit medicinal materials, adding an extraction solvent, mixing, extracting, filtering an extracting solution, and concentrating to obtain the total extract of the cocklebur fruit medicinal materials.

(2) Separation and purification: the total extract of the cocklebur fruit medicinal material is subjected to separation and purification steps to obtain the total glycoside extract of the cocklebur fruit.

The extraction, separation and purification method mentioned here includes the solvent extraction method, macroporous adsorbent resin method, column chromatography and other extraction, separation and purification methods known in the art as mentioned above. Specifically, the method comprises the following steps:

in the first step:

in the solvent extraction method, the extraction solvent includes one or more of water, alcohol, and other organic solvents, the alcohol extraction solvent includes one or more of methanol, ethanol, an ethanol aqueous solution, propanol, butanol, and the like, the other organic solvents include ethyl acetate, acetone, and the like, preferably one or more of water, methanol, ethanol, an ethanol aqueous solution, propanol, butanol, ethyl acetate, acetone, and the like, or a mixed solvent thereof, more preferably one of ethanol, an ethanol aqueous solution, and the like, and particularly preferably 60% to 90% ethanol aqueous solution.

② in the solvent extraction method, the extraction method is preferably one or more of ultrasonic extraction method, percolation extraction method or heating reflux extraction method, and the like, and further preferably heating reflux extraction method; the number of extraction times may be one or more.

Thirdly, the extracting solution is decompressed and filtered, and concentrated to obtain the total extract of the cocklebur fruit medicinal material, the yield of the total extract of the cocklebur fruit medicinal material is 5.0 to 20 percent, namely the total extract of the cocklebur fruit medicinal material accounts for 5.0 to 20 percent of the total weight of the cocklebur fruit medicinal material.

In the second step:

firstly, the adopted separation and purification method is preferably a solvent extraction method, namely, the total extract of the cocklebur fruit medicinal material is dispersed by water, degreased by petroleum ether and extracted by adopting a proper organic solvent to remove fat-soluble components, so as to obtain an extraction residue; or directly extracting with proper organic solvent to remove liposoluble components to obtain extraction residue; the proper organic solvent can be selected from one or more of chloroform, dichloromethane, diethyl ether or ethyl acetate for extraction; the number of extractions may be one or more.

② in the solvent extraction method, the extraction residue obtained after extracting the fat-soluble component is extracted by n-butyl alcohol, the n-butyl alcohol layer is divided, after adding anhydrous sodium sulfate to dehydrate, the n-butyl alcohol part is decompressed and concentrated, the cocklebur fruit total glycoside extract is obtained. At this time, the yield of the cocklebur fruit total glycoside extract is 1-10%, namely the cocklebur fruit total glycoside extract accounts for 1-10% of the total weight of the cocklebur fruit medicinal material.

③ the adopted separation and purification method can also adopt a macroporous absorbent resin method. The adopted macroporous adsorption resin method can recover the total glycoside extract of the cocklebur fruit medicinal material under reduced pressure until no alcohol smell exists, the filtrate is obtained by filtration, the filtrate is adjusted to the relative density of 1.0-1.1 by distilled water, a macroporous adsorption resin column is put on, water is used for washing off water-soluble impurities, elution is carried out by using an elution solvent such as ethanol water solutions with different concentrations, 5-75% of ethanol elution parts are collected, the eluent rich in phenolic acids and diterpenoid glycoside compounds is obtained, and the total glycoside extract of the cocklebur fruit is obtained by reduced pressure recovery until the extract is dried. At this time, the yield of the cocklebur fruit total glycoside extract is 0.5-10%, namely the cocklebur fruit total glycoside extract accounts for 0.5-10% of the total weight of the cocklebur fruit medicinal material. The preferred elution solvent is 40% strength aqueous ethanol.

The macroporous adsorption resin of the cocklebur fruit total glycoside extract can adopt various types of macroporous adsorption resins, including one or more of nonpolar and low polar resins such as D101, D520, D4006, H103, AB-8, Dianion HP 20 or XAD-7, and the like, and can also be other fillers which can replace the resins in terms of performance, preferably one or more of AB-8, D101 or D520, and the like, and particularly preferably AB-8.

In summary, in the preparation method of the cocklebur fruit total glycoside extract, fresh cocklebur fruit medicinal materials or commercially available medicinal materials are taken as raw materials; pulverizing raw materials, sequentially extracting with one or more of solvent extraction method, solvent extraction method or macroporous adsorbent resin method to obtain crude product of fructus Xanthii total glycoside extract; if one or more of the common drying methods such as reduced pressure concentration drying, spray drying, freeze drying and the like are combined, the extract or the dried product of the crude product of the cocklebur fruit total glycoside extract can be prepared.

If a separation and purification method such as column chromatography is further adopted, phenolic acid compounds, diterpene glycoside compounds and the like in the cocklebur fruit total glycoside extract can be obtained; at the moment, the obtained phenolic acid compounds and diterpenoid glycoside compounds account for 50-100 percent of the content of the cocklebur fruit total glycoside extract, wherein the diterpenoid glycoside compounds account for 0.5-99 percent (W/W) of the content of the cocklebur fruit total glycoside extract, and the phenolic acid accounts for 0.5-99 percent (W/W) of the content of the total glycoside extract.

The following will describe in detail the specific preparation method of various formulations of the cocklebur fruit total glycoside extract by taking several typical formulations as examples.

The powder injection preparation of the invention generally adopts a conventional freeze-drying method, takes water as a solvent, and comprises the following steps: adding excipient into fructus Xanthii total glycoside extract, dissolving in water, adding active carbon, filtering, sterilizing, bottling, half-plugging, freeze drying, and capping. The excipient is selected from one or more of mannitol, hydrolyzed gelatin, glucose, lactose or dextran. Each bottle contains 10-100 mg of cocklebur fruit total glycoside extract.

The powder injection preparation of the invention can also adopt a spray drying method, takes water as a solvent, and comprises the following steps: dissolving fructus Xanthii total glycoside extract with or without excipient (the same as above), adding active carbon, filtering for sterilization, spray drying, aseptically packaging, and capping. Each bottle contains 10-100 mg of cocklebur fruit total glycoside extract.

When the small injection is prepared, water for injection is used as a solvent for preparation, and a proper amount of auxiliary materials can be added, wherein the auxiliary materials are selected from one or more of ethanol, propylene glycol, glycerol, polyethylene glycol, benzyl benzoate or dimethylacetamide. Each branch contains 10-100 mg of cocklebur fruit total glycoside extract.

The glucose infusion or sodium chloride infusion is prepared by taking water for injection as a solvent and adding a proper amount of glucose or sodium chloride, and also can be prepared by adding a proper amount of auxiliary materials, wherein the auxiliary materials are selected from one or more of ethanol, propylene glycol, glycerol, polyethylene glycol, benzyl benzoate or dimethylacetamide. Each bottle contains 10-100 mg of cocklebur fruit total glycoside extract.

The invention can be prepared into tablets, capsules, granules, oral liquid and other oral preparations, and the auxiliary materials can be lactose, starch, dextrin, stearate and the like, and are prepared according to the conventional technology.

In the present invention, the above-described embodiments and the following examples are provided to better illustrate the present invention and are not intended to limit the scope of the present invention.

The present invention will be described in detail by examples.

Example 1 preparation of cocklebur fruit Total glycosides extract 1

2.5Kg of fructus xanthil dried medicinal material is extracted by 50 liters of 90 percent ethanol solution under reflux for 2 times, each time lasts for 2 hours, the two extracting solutions are combined, the extract is concentrated under reduced pressure to obtain 350g of extract, after the extract is dispersed by 2 liters of water, the extract is sequentially extracted and degreased by 6 liters of petroleum ether for three times, and the impurities are respectively extracted and removed by 6 liters of chloroform and 6 liters of ethyl acetate for three times; extracting the extraction residue with 6L n-butanol for three times, mixing extractive solutions, and freeze drying to obtain 21g of fructus Xanthii total glycoside extract.

Example 2 preparation of cocklebur fruit Total glycosides extract 2

2.5Kg of fructus xanthil dried medicinal material is ultrasonically extracted for 2 times with 50L of 70% ethanol solution, each time lasts for 2 hours, the two extracting solutions are combined, the extract is decompressed and concentrated to obtain 208g of extract, the extract is dispersed by 2L of water, extracted and degreased by 6L of petroleum ether for three times in sequence, and extracted and decontaminated by 6L of chloroform and 6L of ethyl acetate for three times respectively; extracting the extraction residue with 6L n-butanol for three times, mixing extractive solutions, and recovering under reduced pressure to dry to obtain fructus Xanthii total glycoside extract 17 g.

Example 3 preparation of cocklebur fruit Total glycosides extract 3

2.5Kg of fructus xanthil dried medicinal material is percolated and extracted for 2 times by 50L of 75 percent ethanol solution, each time lasts for 24 hours, the two extracting solutions are combined, the extract is concentrated under reduced pressure to obtain 285g of extract, after being dispersed by 2L of water, the extract is sequentially extracted and degreased by 6L of petroleum ether for three times, and the impurities are respectively extracted and removed by 6L of chloroform and 6L of ethyl acetate for three times; extracting the extraction residue with 6L n-butanol for three times, mixing extractive solutions, and recovering under reduced pressure to dry to obtain fructus Xanthii total glycoside extract 19 g.

Example 4 preparation of cocklebur fruit Total glycosides extract 4

2.5Kg of fructus xanthil dried medicinal material is extracted by 50 liters of 75 percent ethanol solution under reflux for 2 times, each time lasts for 2 hours, the two extracting solutions are combined, the pressure is reduced and recovered until no alcohol smell exists, the filtrate is obtained by filtration, the relative density of the filtrate is adjusted to 1.05 by distilled water, the filtrate is put on a D101 macroporous adsorbent resin column, water-soluble impurities are washed away by water, then ethanol with different concentrations is used for elution, the elution part of 50 percent ethanol is collected, the elution liquid rich in phenolic acid glycosides and diterpene glycosides is obtained, the pressure is reduced and recovered to be dry, and 15g of fructus xanthil total glycoside extract is obtained.

Example 5 preparation of cocklebur fruit Total glycosides extract 5

2.5Kg of fructus xanthil dried medicinal material is extracted by 50 liters of 75 percent ethanol solution under reflux for 2 times, each time lasts for 2 hours, the two extracting solutions are combined, the pressure is reduced and recovered until no alcohol smell exists, the filtrate is obtained by filtration, the filtrate is adjusted to the relative density of 1.05 by distilled water, an AB-8 macroporous absorbent resin column is used, water-soluble impurities are washed away by water, ethanol with different concentrations is used for elution, 50 percent ethanol elution parts are collected, the eluent rich in phenolic acid glycosides and diterpene glycoside compounds is obtained, the pressure is reduced and recovered to be dry, and 20g of fructus xanthil total glycoside extract is obtained.

Example 6 preparation of nose drops of cocklebur fruit Total glycoside extract

Taking 45g of the obtained cocklebur fruit total glycoside extract, adding 20g of pig gall cream, 0.5g of chlorphenamine maleate and 8g of sodium houttuyfonate, adding distilled water for diluting by 10 times, filtering out insoluble substances, bottling, sterilizing and sealing to obtain the cocklebur fruit total glycoside extract nasal drops.

Example 7 preparation of Xanthium sibiricum Total glycosides extract drop capsules

Taking 40g of the obtained cocklebur fruit total glycoside extract, adding 15g of pig bile paste, 0.3g of chlorphenamine maleate and 6g of sodium houttuyfonate, adding 40-60% of fillers such as starch, dextrin, carboxymethyl starch, calcium hydrophosphate and the like, 0.5-10% of disintegrant such as sodium carboxymethyl cellulose and hydroxypropyl cellulose, preparing into granules, drying, adding glidant such as magnesium stearate and silica gel, drying, and filling into capsules to obtain the product, namely the cocklebur fruit total glycoside extract capsule.

Example 8 preparation of Capsule of Xanthium sibiricum Total glycosides extract

(1) Prescription

(2) The preparation process comprises the steps of respectively taking the raw material medicine cocklebur fruit total glycoside extract and other auxiliary materials in the prescription according to the prescription, respectively sieving the raw material medicine cocklebur fruit total glycoside extract and the auxiliary materials in the prescription through a 100-mesh sieve, drying at 60 ℃, weighing the prescription amount of the cocklebur fruit total glycoside extract, uniformly mixing the cocklebur fruit total glycoside extract, microcrystalline cellulose and sodium carboxymethyl starch in an equal incremental method, preparing a soft material by using a proper amount of absolute ethyl alcohol, granulating through a 30-mesh sieve, drying at 50-60 ℃ for 2 hours, granulating through a 30-mesh sieve, adding the prescription amount of.

Example 9 preparation of powder injection of cocklebur fruit Total glycosides extract

Taking 30g of cocklebur fruit total glycoside extract, adding 30g of dextran, adding 500ml of water for injection, and stirring to dissolve the dextran; adding water for injection to 2000ml, adding 3.0g of active carbon for injection, and fully stirring for 30 minutes; decarbonizing and filtering; filtering with 0.22 μm microporous membrane; filling into sterile penicillin bottles with 2ml per bottle, and half-rolling a stopper; freeze drying, and then plugging and capping.

Example 10 preparation of powder injection of xanthium sibiricum total glycoside extract

Taking 60g of cocklebur fruit total glycoside extract, adding 500ml of water for injection, and stirring to dissolve the cocklebur fruit total glycoside extract; adding water for injection to 1000ml, adding 1g of active carbon for injection, and fully stirring for 30 minutes; decarbonizing and filtering; filtering with 0.22 μm microporous membrane; freeze drying to obtain sterile powder, and subpackaging into 1000 bottles.

Example 11 preparation of powder injection of cocklebur fruit Total glycosides extract

Taking 40g of cocklebur fruit total glycoside extract, adding 50g of lactose and 100ml of water for injection, and stirring to dissolve; adding water for injection to 1000ml, adding 1.5g of active carbon for injection, and fully stirring for 30 minutes; decarbonizing and filtering; filtering with 0.22 μm microporous membrane; spray drying to obtain sterile powder, and subpackaging into 1000 bottles.

EXAMPLE 12 preparation of powder injection of cocklebur fruit Total glycosides extract

(1) Prescription

(2) Preparation process

Weighing the cocklebur fruit total glycoside extract according to the prescription amount, adding the cocklebur fruit total glycoside extract into a proper amount of water for injection, and stirring to dissolve the cocklebur fruit total glycoside extract; adding mannitol in a prescription amount, stirring to completely dissolve, and adding water for injection to the full amount; adding 0.1% of liquid amount of needle activated carbon, and fully stirring for 30 minutes; decarbonizing and filtering; filtering with 0.22 μm microporous membrane; filling and half-rolling stopper; freeze drying, and capping. 9735 bottles are prepared, and the yield is 97.35%.

EXAMPLE 13 preparation of a Small injection containing Xanthium sibiricum Total glycosides extract

Taking 5g of cocklebur fruit total glycoside extract, adding 100ml of water for injection, and stirring to dissolve the cocklebur fruit total glycoside extract; adding water for injection to 1000ml, and filtering with 0.22 μm microporous membrane; subpackaging and encapsulating, 10ml per bottle, and sterilizing.

EXAMPLE 14 preparation of a Small injection containing Xanthium sibiricum Total glycosides extract

Taking 10g of cocklebur fruit total glycoside extract, adding 30g of propylene glycol and 200ml of water for injection, and stirring to dissolve the cocklebur fruit total glycoside extract; adding water for injection to 1000ml, adding 1.5g of active carbon for injection, and fully stirring for 30 minutes; decarbonizing and filtering; filtration through a 0.22 μm microfiltration membrane: subpackaging and encapsulating, 5ml per bottle, and sterilizing.

Example 15 preparation of glucose infusion solution of Xanthium sibiricum Total glycosides extract

Taking 2g of cocklebur fruit total glycoside extract, adding 10g of polyethylene glycol, adding 500g of glucose, adding 2000ml of water for injection, and stirring to dissolve; adding water for injection to 5000 ml; filtering with 0.22 μm microporous membrane; packaging, bottling, and sterilizing, wherein each bottle contains 100 ml.

Example 16 preparation of glucose infusion solution of Xanthium sibiricum Total glycosides extract

Taking 2g of cocklebur fruit total glycoside extract, adding 250g of glucose and 1000ml of water for injection, and stirring to dissolve the cocklebur fruit total glycoside extract; adding water for injection to 5000 ml; filtering with 0.22 μm microporous membrane; subpackaging and encapsulating, 250ml per bottle, and sterilizing.

Example 17 preparation of infusion solution of Soujishi Total glycosides extract sodium chloride

Taking 1g of cocklebur fruit total glycoside extract, adding 90g of sodium chloride and 1000ml of water for injection, and stirring to dissolve the cocklebur fruit total glycoside extract; adding water for injection to 10000 ml; filtering with 0.22 μm microporous membrane; subpackaging and encapsulating, 250ml per bottle, and sterilizing.

EXAMPLE 18 preparation of tablets of the Total glycosides extract of Xanthium Sibiricum

Taking 80g of cocklebur fruit total glycoside extract, 80g of microcrystalline cellulose, 15g of lactose and 60g of pregelatinized starch, sieving, uniformly mixing, preparing a soft material by using a proper amount of 10% PVP ethanol solution, granulating, drying, adding 3g of magnesium stearate, finishing granules, tabletting and preparing into 1000 tablets.

EXAMPLE 19 preparation of tablets of the Total glycosides extract of Xanthium Sibiricum

(1) Prescription

(2) Preparation process

The main medicine and the auxiliary materials are respectively weighed according to the prescription, the main medicine and the auxiliary materials are uniformly mixed according to an equivalent progressive method, the processes of soft material preparation, granulation, drying, granule straightening and the like are carried out according to the prescription process, a single-punch tablet machine and a 10.5mm shallow concave punch die are used for tabletting after the tablet weight is calculated, the hardness of a bare chip is controlled to be 5-7 kg, tablets 9698 are prepared, and the yield is 96.98%.

Coating by adopting a rolling spraying method, wherein the coating process comprises the following steps:

preparing a coating solution: gastric soluble film coating material: 85G61235 from Shanghai Kalekang coating technology Co., Ltd

The coating process comprises the following steps: putting a bare chip (with the hardness of 5 kg-7 kg) to be coated into a coating pan, starting a stirring device and an air blast heating device, opening a spray gun to aim at 1/3 of a tablet bed to spray coating liquid for coating when the temperature of the bare chip rises to 40 ℃, controlling the temperature of the tablet bed to be 38-42 ℃, the gas pressure to be 6kg, the flow rate of the coating liquid to be 50mL/min, and the weight of a coating film to be 3% of that of the coating tablet.

EXAMPLE 20 analgesic Effect of Xanthium sibiricum Total glycosides extract

Taking appropriate amount of fructus Xanthii total glycoside extract (CE for short), and observing analgesic effect of fructus Xanthii total glycoside by adopting mouse writhing reaction model induced by acetic acid.

(1) Experimental methods

60 mice of 20 +/-2 g, each half of male and female, were randomly grouped. The composition was administered continuously for 3 days at the doses shown in Table 6, and was intraperitoneally injected with 0.7% glacial acetic acid solution (0.1ml/10g mouse) 30min before the last administration, and the number of writhing of the mouse was observed and recorded within 10min after 5min, and the results are shown in Table 3.

(2) Results of the experiment

The results in Table 3 show that each group of CE can significantly reduce the frequency of writhing reaction of mice caused by acetic acid, and has analgesic effect.

TABLE 3 Effect of CE on acid-induced writhing response in mice (X + -SD, n ═ 10)

Compared with the group of the normal saline solution,^*P<0.05，^**P<0.01

example 21 therapeutic Effect of Xanthium sibiricum Total glycosides extract on Experimental sinusitis

Taking a proper amount of the obtained cocklebur fruit total glycoside extract (CE for short), adopting a rabbit experimental nasosinusitis model, and observing the influence of the cocklebur fruit total glycoside on the rabbit nasal mucosa histomorphology by using a light mirror.

(1) Experimental methods

56 healthy New Zealand white rabbits with the weight of about 2.5kg are selected, and the rabbits are divided into groups in which the male rabbit and the female rabbit are half female rabbit and half male rabbit. The first day, general anesthesia, the left maxillary sinus ostium was opened, the unilateral sinus ostium was blocked with gelatin sponge, and surgical suturing was performed. The whole process is carried out according to aseptic operation. Injecting 108CFU/ml Streptococcus pneumoniae suspension 0.5ml the next day, continuously adding medicine according to the dosage in Table 1 for 10 days, separating and culturing cells after the last administration, taking materials, and making slices. The following indices were observed under a light microscope: the thickness of the nasal mucosa is measured by an ocular micrometer to be convenient for positioning comparison, and the thickness (including an epithelial layer and an inherent membrane) from the vertex of the mucosa bulge of each specimen to the lateral edge of the nasal septum cartilage is taken as the thickness of the mucosa; the number of glands in the mucosa lamina propria is based on the average number of glands under the bilateral mucosa bulge; ③ the number of the goblet cells of the mucosa epithelium is determined by randomly counting the average number of the total number of the 5 goblet cells with high visual field; fourthly, the number of lymph nodules in the mucosa lamina propria is the total number of lymph nodules in the section.

(2) Results of the experiment

The morphological changes of the nasal mucosa tissues of each group are observed and analyzed, and the results are shown in a table 4.

The results in tables 3 and 4 show that the CE medium and high dose groups obviously reduce the infiltration of nasal mucosa cells, eliminate the edema of nasal mucosa, inhibit the proliferation of connective tissue under the mucosa and reduce the hyperplasia of glands in epithelial goblet cells of the nasal mucosa and the inherent layer of the mucosa, and the indexes of the tissue morphology are all stronger than those of the levofloxacin group, and the high dose group is stronger than that of the nasosinusitis group.

TABLE 4 Effect of CE on rabbit sinus mucosal histomorphometric indices (X + -SD, n ═ 8)

In comparison with the set of models,^*P<0.05，^**P<0.01

example 22 influenza Virus inhibition by Total glycosides extract of Xanthium Sibiricum

(1) Experimental methods

(mdck (madin darby Canin kidney) cell passaging and influenza virus (influenza a 1 virus) culture: the method is carried out according to the literature (Guoguang, Chengxin; 1997).

② cytotoxicity test: adding the sample into a cell plate which has grown into a monolayer of cells at a rate of 0.1 mL/hole, adding a cell maintenance solution to the concentration of 1 mL/hole, culturing at 37 ℃ in a 5% CO2 incubator for 72h, and observing cytopathic effect. And simultaneously setting MDCK cell control. The experiment was repeated 2 times. The results show that: the samples did not produce non-specific cytopathic effects (CPE) on MDCK cells.

③ anti-influenza virus test of the sample: cells were cultured in 96-well MDCK cell culture plates, and a cell control group, a virus control group, a positive control group, and a test group were provided, respectively. Influenza A1 virus was added to the virus control and test groups, adsorbed at 37 ℃ for 2h, and virus aspirated. Adding samples with different concentrations respectivelyBy each test group, cultured at 37 ℃ for 3 days with 5% CO2, test results were observed, and 50% Inhibitory Concentration (IC) of different drugs against viruses was calculated₅₀)。

(2) Experimental results (see: Table 5)

The experimental result shows that the cocklebur fruit total glycoside extract has obvious inhibition effect on influenza virus.

TABLE 5 inhibitory Effect of cocklebur fruit Total glycosides extract on influenza Virus

Group of	IC<sub>50</sub>(mmol/L)
Group of	IC<sub>50</sub>(mmol/L)	CE	4.2
Ribavirin	3.9	CE	4.2

Claims

1. The application of the cocklebur fruit total glycoside extract in preparing anti-inflammatory reaction products, wherein the anti-inflammatory reaction products refer to products for resisting viruses and related diseases, products for diagnosing, detecting, treating or researching acute and chronic rhinitis and related diseases, or one or more products for diagnosing, detecting, treating or researching nasosinusitis and related diseases;

the total glycoside extract is a total extract containing diterpene glycoside compounds, phenolic acid compounds and other components;

the content ranges of the diterpene glycoside compounds, the phenolic acid compounds and other components in the total glycoside extract are respectively 0.5-99% (weight percentage), 0.5-99% and less than 0.5%;

the chemical structure general formula of the diterpene glycoside compounds is as follows:

wherein R is₁Is one of hydrogen, methyl or carboxyl; r₂、R₃Is one of hydrogen, sulfonic group, acetyl, propionyl or dimethylpropionyl;

the chemical structural general formula of the phenolic acid compound is as follows:

2. The use of the extract of cocklebur fruit total glycosides according to claim 1, characterized in that the anti-inflammatory reaction product is one or more selected from the group consisting of a drug, a reagent, a food or a beverage.

3. The use of the fructus xanthil total glycoside extract as claimed in claim 2, wherein the anti-inflammatory reaction product is a medicament for diagnosing, detecting, treating or studying acute and chronic rhinitis and related disorders, or a medicament for diagnosing, detecting, treating or studying sinusitis and related disorders.

4. The use of the fructus xanthil total glycoside extract as claimed in claim 1, wherein the content of diterpene glycosides and phenolic acids in the total glycoside extract is 5% -95% and 5% -95%, respectively.

5. The use of the fructus xanthil total glycoside extract as claimed in claim 1, wherein the diterpene glycosides are one or more compounds selected from the group consisting of:

when R is₁＝COOH，R₂＝R₃When H, represents diterpene glycoside compound a;

6. The use of the cocklebur fruit total glycoside extract according to claim 1, characterized in that the phenolic acid compound is one or more of the following compounds:

when R is₁＝K，R₂＝R₄＝H，R₃When caffeoyl, it represents a phenolic acid compound D.

7. The use of the cocklebur fruit total glycoside extract according to claim 1, characterized in that the preparation method of the cocklebur fruit total glycoside extract comprises the following steps:

8. The use of the extract of cocklebur fruit total glycosides according to claim 7, characterized in that the extraction method is all available methods known in the art including solvent extraction.

9. The use of the extract of cocklebur fruit total glycosides according to claim 8, characterized in that the extraction method is a solvent extraction method.

10. The use of the cocklebur fruit total glycoside extract according to claim 9, characterized in that the solvent extraction method comprises one or more of the commonly used ultrasonic extraction method, maceration extraction method, percolation extraction method, decoction extraction method, heating reflux extraction method or continuous reflux extraction method; the number of extraction is one or more.

11. The use of the cocklebur fruit total glycoside extract according to claim 10, characterized in that the solvent extraction method comprises one of ultrasonic extraction, percolation extraction or heating reflux extraction; the number of extractions is multiple.

12. The use of the cocklebur fruit total glycoside extract according to claim 11, characterized in that the extraction solvent used in the solvent extraction method is one or more of three types of extraction solvents including common aqueous reagents, hydrophilic organic solvents or lipophilic organic solvents.

13. The use of the extract of cocklebur fruit total glycosides according to claim 12, characterized in that the aqueous agent is one selected from the group consisting of water, acid water or alkaline water; the hydrophilic organic solvent is one or more of ethanol, ethanol water solution or methanol; the lipophilic organic solvent is one or more selected from petroleum ether, chloroform, diethyl ether, ethyl acetate, dichloromethane or dichloroethane.

14. The use of the fructus xanthil total glycoside extract as claimed in claim 7, wherein the extraction method comprises: taking a plurality of cocklebur fruit medicinal materials, adding an extraction solvent for mixing, extracting, filtering an extracting solution, and concentrating to obtain the total extract of the cocklebur fruit medicinal materials.

15. The use of the extract of cocklebur fruit total glycosides according to claim 14, characterized in that the extraction solvent is one or more of water, alcohols or other organic solvents.

16. The use of the cocklebur fruit total glycoside extract according to claim 15, characterized in that the alcohol extraction solvent comprises one or more of methanol, ethanol water solution, propanol or butanol, and the other organic solvent comprises one or more of ethyl acetate or acetone.

17. The use of the cocklebur fruit total glycoside extract according to claim 16, characterized in that the alcohol extraction solvent is 60% -90% ethanol aqueous solution.

18. The use of the extract of cocklebur fruit total glycosides according to claim 7, characterized in that the separation and purification method comprises one or more of solvent separation, solvent extraction, macroporous adsorbent resin, precipitation, salting out, column chromatography, or crystallization and recrystallization and fractional crystallization.

19. The use of the extract of cocklebur fruit total glycosides according to claim 18, characterized in that the solvent extraction method comprises one or more of extraction method, countercurrent continuous extraction method, countercurrent distribution method or droplet countercurrent distribution method.

20. The use of the cocklebur fruit total glycoside extract according to claim 19, characterized in that the solvent extraction method comprises the following steps:

(1) dispersing the fructus Xanthii total extract with water, defatting with petroleum ether, and extracting with appropriate organic solvent to remove liposoluble components to obtain extraction residue; the number of extractions may be one or more.

(2) Extracting the extraction residue with n-butanol, separating n-butanol layer, adding anhydrous sodium sulfate for dehydration, and concentrating n-butanol part under reduced pressure to obtain fructus Xanthii total glycoside extract.

21. The use of the cocklebur fruit total glycoside extract according to claim 19, characterized in that the solvent extraction method comprises the following steps: directly extracting the total extract of the fructus xanthil with a proper organic solvent to remove fat-soluble components to obtain an extraction residue; the number of extractions may be one or more.

22. The use of the cocklebur fruit total glycoside extract according to claim 21, characterized in that the suitable organic solvent is one or more selected from the group consisting of chloroform, dichloromethane, diethyl ether or ethyl acetate.

23. The use of the cocklebur fruit total glycoside extract according to claim 18, characterized in that the operation of the macroporous adsorbent resin method comprises the following steps:

(1) recovering fructus Xanthii total glycosides extract under reduced pressure until no alcohol smell exists, and filtering to obtain filtrate;

(2) adjusting the relative density of the filtrate to 1.0-1.1 by using distilled water, putting the filtrate on a macroporous adsorption resin column, and washing away water-soluble impurities by using water;

(3) eluting with an elution solvent, and collecting the elution part of 5-75% ethanol to obtain an eluent rich in phenolic acids and diterpene glycosides;

(4) recovering under reduced pressure to dry to obtain fructus Xanthii total glycoside extract.

24. The use of the cocklebur fruit total glycoside extract according to claim 23, characterized in that the elution solvent is an aqueous solution of ethanol of various concentrations.

25. The use of the cocklebur fruit total glycoside extract according to claim 24, characterized in that the elution solvent is 40% aqueous ethanol.

26. The use of the extract of cocklebur fruit total glycosides according to claim 23, characterized in that the macroporous adsorbent resin comprises a plurality of types of macroporous adsorbent resin.

27. The use of the extract of cocklebur fruit total glycosides according to claim 26, characterized in that the macroporous adsorbent resin is one comprising a non-polar resin, a weakly polar resin or other fillers capable of substituting the above-mentioned resins in their properties.

28. The use of the extract of cocklebur fruit total glycosides according to claim 27, wherein the non-polar resin and the less polar resin is one of D101, D520, D4006, H103, AB-8, Dianion HP 20 or XAD-7.

29. The use of the extract of cocklebur fruit total glycosides according to claim 7, characterized by further comprising a drying step.

30. The use of the extract of cocklebur fruit total glycosides according to claim 29, wherein the drying method comprises one or more of an atmospheric drying method, a reduced pressure drying method, a vacuum drying method, a spray drying method, a freeze drying method, a far infrared heat drying method, or a microwave drying method.