CN1994352A - Use of total glucosides extract of cocklebur fruit in preparation of product for resisting inflammatory reaction - Google Patents

Abstract

The invention relates to the application of the extract of total glycosides of the traditional Chinese medicine Xanthium fructus. The total glycoside extract includes total extracts of diterpene glycosides, phenolic acid compounds and other components, and the content ranges are respectively 0.5%-99%, 0.5%-99% and less than 0.5%. The total glycoside extract can be obtained by adopting conventional extraction, separation and purification methods and steps in this field. The extraction method generally adopts solvent extraction method, etc., and the separation and purification method generally adopts solvent extraction method, macroporous adsorption resin method, etc. The total glycoside extract has strong pharmacological effects, obvious effects of anti-inflammation, anti-virus, analgesia, and treatment of acute and chronic rhinitis, sinusitis and related diseases, and has simple preparation process, high yield and stable chemical properties. Acute and chronic sinusitis is a common and frequently-occurring disease in clinical practice, and the incidence rate is constantly rising. The total glycoside extract has rich sources of medicinal materials, so it can be further developed into a new drug with clinical application prospects.

Description

Application of Xanthium glycosides extract in the preparation of anti-inflammatory reaction products

This application is a divisional application of the invention patent application "Usage of Total Glycosides Extract of Xanthia Fructus for the Preparation of Anti-inflammatory Reaction Products". The filing date of the original application is March 28, 2005, and the application number is 200510024693.9. It is an invention and creation The name is: the use of the total glycosides extract of Xanthium Fructus for preparing anti-inflammatory reaction products.

technical field

The invention relates to the technical fields of medicine, food and beverage, in particular to the use of Chinese medicine extracts, and more specifically to the use of Chinese medicine cocklebur glycosides extracts for preparing anti-inflammatory reaction products.

Background technique

(1) Research overview of Fructus Xanthii

1. Overview of plant resources and applications

Fructus Xanthii (Fructus Xanthii), is the dry and mature fruit with involucre of Xanthium sibirium Patr. Cangzike, etc., are Chinese Pharmacopoeia varieties, which are produced in most parts of China and are extremely rich in resources. Traditional Chinese medicine believes that it has a bitter taste and warm nature, and it belongs to the lung and liver meridians. It has the effects of sweating, dispelling rheumatism, clearing the nose, and relieving pain. It is traditionally used to treat wind-cold headache, runny nose, rubella itching, arthralgia due to dampness, etc. It is clinically used to treat acute and chronic rhinitis, sinusitis, bronchitis, low back pain, rubella, scabies and other skin diseases (State Administration of Traditional Chinese Medicine. Chinese Materia Medica [M]. Seventh volume. Shanghai: Shanghai Science and Technology Press, 1999:1013-1016).

2. Chemical composition

Studies have shown that the chemical components contained in Xanthium plants are mainly sesquiterpene lactones, volatile oils and water-soluble glycosides, which are described as follows:

(1) Sesquiterpene lactone compounds Xanthium plants mainly contain guaiac-type and split-guaiac-type lactone compounds, mainly including xanthinin (i.e. cryptoxanthin), xanthin ( Xanthumin, that is, xanthumin, is a stereoisomer of xanthumin), xanthumin (xanthin, that is, xanthin) and their derivatives. In addition, there are xanthanol, xanthinosin, xantholide A and xantholide B in X.canadense, ziniolide and lasidiol p-methoxybenzoate in X.catharticum, and pungiolide A and pungiolide B in X.pungens (Salinas-A, De-Ruiz-RE, Ruiz-SO. Sterols, flavonoids and sesquiterpenic lactones from Xanthiun spinosum (Asteraceae). Acta Farm Bonaerense, 1998, 17:297; and Kis-I, Racz-G. Glycoside contents of sitosterol from leaves of Xanthiun spinosum and X. italicum. Farmacia Bucharest, 1988, 36(1): 55; and Ahmed-AA. New vomifoliol derivative from Xanthiun pungens. Pharmazie, 1990, 45: 698).

(2) Volatile oil Ahijja-MM and others extracted the volatile oil from the leaves of Xanthiunstmmarium (Linn.) by steam distillation (Ahijja-MM, Nigam-SS. Chemical examination of the essential oil from the leaves of Xanthiunstmmarium(Linn.).Flavour Ind , 1970, 1:627), found to contain d-limonene (d-limonene, 35%), cymene (p-cymene, 5%), β-syringene (beta-caryophyllene, 6%), terpineol Alkene (terpinolene, 7.3%), 1-α-ionone (1-alpha-ionone, 10.9%), d-high terpene alcohol (d-carveol, 25%) and α-pinene (alpha-pinene, 0.4% ). Taher-HA et al analyzed the volatile oil components in X.cavanillesii (Taher-HA, Ubiergo-GO, Talenti-ECJ.Constituents of the essential oil of Xanthiun cavanillesii.J Nat Prod, 1985,48:857.), which contains mono Terpenoids 55.2%, Oxidized Monoterpenoids 14.2%, Sesquiterpenoids 11.3%. Guo Yahong and others analyzed 17 chemical components from the volatile oil of Fructus Xanthii and calculated their relative percentages (Guo Yahong, Li Jiashi, Pan Jiongguang, etc. Research on volatile oil in Fructus Xanthii. Chinese Journal of Traditional Chinese Medicine, 1994, 19 (4) :235).

(3) Water-soluble glycosides As early as the 1960s, Song Zhenyu and others had isolated a substance with glycoside properties (tentatively named AA2) from the water infusion of Xanthium, which may be the main toxicity of Xanthium. Components (Song Zhenyu, Zhang Lingyun, Xie Mingming, etc. The toxic components and pharmacological effects of Xanthium Fructus. Acta Pharmaceutica Sinica, 1962, 9 (11): 678). MacLeod-JK et al. extracted two water-soluble toxic kaurene glycosides (kaureneglycosides) from the thorn fruit of the spicy Xanthium X.pungens (MacLeod-JK, Moeller-PD, Franke-FP.Two toxic glycosides from The burrs of Xanthiun pungens. J Nat Prod, 1990, 53: 451), identified by two-dimensional nuclear magnetic resonance and mass spectrometry, confirmed that its structure is a derivative of carboxy atractyloside. In addition, the same water-soluble toxic glycoside was extracted from Xanthium and Mongolian Xanthium, which was identified as atracyloside (Wang Suxian, Ren Lijuan, Sun Zeren, etc. Toxic components in Xanthium mongolica seeds. Chinese herbal medicine , 1983, 14(12): 1).

(4) Other Cocklebur also contains cockleburin (strumaroside, namely β-sitosterol glucoside), 8-(Δ3-prenyl)-5,7,3′,4′-tetrahydroxyflavone, and Caffeic acid and 1,4-dicaffeoylquinic acid. In addition, it still contains alkaloids, tannins, chalcone derivatives, triterpenoids, glucose, fructose, amino acids, tartaric acid, etc.

3. Pharmacological activity

(1) Antimicrobial effect Xanthium decoction has a strong inhibitory effect on various microorganisms such as Staphylococcus aureus, Bacillus anthracis, pneumococcus, Streptococcus B and Bacillus diphtheria in vitro. Xanthia acetone or ethanol extract has antifungal effect on Trichophyton rubrum, and its aqueous extract has antifungal effect on Trichophyton violaceum. Zhang Zheng et al. researched in 1988 and confirmed that cocklebur decoction had a direct inhibitory rate of 25% to 50% on DNA polysaccharides of hepatitis B virus in vitro, indicating that it has anti-hepatitis virus effects (Zhang Zheng, Xu Xiangdong, Du Shaocai, etc. Experimental research on 60 kinds of Chinese herbal medicines against hepatitis B virus. Journal of Beijing Medical University, 1988, 20(3): 211).

(2) Effects on the cardiovascular system Xanthium decoction can inhibit the isolated frog heart and guinea pig heart, slow down the heart rate, weaken the contraction force of the heart, and expand the rabbit ear blood vessels, while the frog blood vessels first expand and then expand Contraction, cocklebur injection, can make the blood pressure of rabbits and dogs drop temporarily. The glycoside component AA2 has a mild hypotensive effect on rats, and can enhance vascular permeability (Song Zhenyu, Zhang Lingyun, Xie Zhiming, etc. Toxic components and pharmacological effects of Xanthium Fructus Xanthii. Acta Pharmacologica Sinica, 1962, 9 (11 ): 678).

(3) Effects on the blood system Xanthium extract (equivalent to crude drug 0.2g/ml) can significantly prolong the time for bovine thrombin to aggregate human fibrinogen, and has obvious antithrombin effect. The methanol extract of Xanthium can quickly restore the reduction of rabbit cholesterol and triglycerides caused by fasting, and can also increase the content of phospholipids to a certain extent.

(4) Effects on immune function Xanthium can significantly inhibit the cellular immunity and humoral immunity of C57/BL purebred mice. Fructus Xanthii reduces the number of helper T cells (abbreviation: TH) and suppressor T cells (abbreviation: TS), and reduces the ratio of TH/TS. Fructus Xanthii can significantly reduce the β-endorphin in the hypothalamus and plasma. In addition, Xanthium can still reduce interleukin-2 (abbreviation: IL-2) activity and IL-2 receptor content, can significantly reduce the release of intracellular histamine, this is Xanthium can be used to treat allergic One of the mechanisms of the disease (Wang Longmei, Fu Huidi, Zhou Zhilan. Effects of Lycium barbarum, Atractylodes macrocephala, Asarum, and Xanthium on the expression of interleukin-2 receptor. Chinese Journal of Clinical Pharmacy, 2000, 9(3): 172) .

(5) Antioxidant effect In 1994, Fan Jingpo studied the antioxidative effect of Fructus Xanthii, and showed that Fructus Xanthii decoction 0.5g of crude drug/gavage, once a day for 10 days, can effectively reduce lipid peroxidation It can reduce the content of lipid peroxide (abbreviation: LPO) in tissue, and has a tendency to increase the activity of superoxide dismutase (abbreviation: SOD), indicating that cocklebur can enhance the body's ability to scavenge free radicals and reduce the impact of free radicals on the body. (Fan Jingpo. Effects of Xanthium, Asarum, Lycium barbarum, and Atractylodes macrocephala on free radical metabolism in mouse tissues. Chinese Medicine Information, 1994, (2): 48).

(6) Anti-inflammatory and analgesic effects Liu Qingzeng and others described the pharmacological activity of cocklebur extract in 1988 (Liu Qingzeng, Wang Jinlan. Some progress in the study of pharmacological effects of traditional Chinese medicine in Japan in recent years. Pharmacology and Clinic of Chinese Medicine, 1988, 4 (2 ): 50). Intraperitoneal injection of 250mg/kg methanol extract of Xanthii can inhibit carrageenan foot swelling in rats by 30%-60%; subcutaneous injection of 1000mg/kg can inhibit acetic acid writhing reaction in mice by 10% ~30%, indicating that this product has certain anti-inflammatory and analgesic effects. Modified Xanthium Pills have good anti-inflammatory effects, including: can significantly reduce the amount of Evans blue exudation in the abdominal cavity of mice caused by acetic acid, and can significantly reduce the swelling degree of mouse ear inflammation caused by xylene. It still has a good analgesic effect and can significantly prolong the time of writhing reaction in mice induced by acetic acid (Ma Ping, Li Hong. Research Progress of Xanthium Fructus. Chinese Herbal Medicine, 1999, 30(8): 634).

(7) Other studies have shown that the water extract of Xanthium semen can inhibit cervical cancer cells by 50% to 70% in vitro; Glutaric acid coenzyme A (abbreviation: HMG-Co-A), calcium channel blocker receptors and cholecystokinin have different degrees of inhibition.

4. Clinical application of single and compound prescriptions (Li Hong, Zhou Mou, pharmacological effects and clinical research progress of cocklebur and compound preparations. Journal of Shanxi Medical University, 2004, 35(3): 313-314)

(1) Treatment of nasal diseases

Treatment of rhinitis Chronic rhinitis is a common disease, frequently-occurring disease. Someone made additions and subtractions on the basis of the original Cangerzi powder, and randomly divided 96 patients into a treatment group and a control group. One month was a course of treatment. According to clinical observation, there was no significant difference between the two groups. Erzi Pill is similar to Biyan Pill in treating chronic rhinitis and slightly better than Biyan Pill.

Treatment of sinusitis Using Huangqin Decoction and Cangerzi Powder to treat chronic sinusitis, 108 cases were clinically observed, and the total effective rate was 94.4%. Generally, the effect can be seen after a course of treatment. Combined use of cocklebur seed mixture and astemizole in the treatment of chronic sinusitis in children has a significant effect. Fructus Xanthii is roasted into brown and ground into powder or powdered into honeyed pills to treat allergic rhinitis. Most patients' symptoms disappear or improve, or the attack decreases. 67 cases of chronic sinusitis were also treated with the ancient formula Xanthium powder plus myrobalan and earthworm, 42 cases were cured, 20 cases improved, and 5 cases were ineffective, with a total effective rate of 92.54%.

Treatment of Biyuan Chinese medicine classifies acute suppurative sinusitis as Biyuan. 200 cases of acute suppurative sinusitis were treated with Xanthium mixture, 100 cases in the treatment group took Xanthium mixture, and 100 cases in the control group were treated with penicillin and compound sulfamethoxazole. The results of clinical treatment show that the total effective rates of the treatment group and the control group are 99% and 95% respectively (P>0.05), and there is no significant difference. Clinically, Cang and Xin decoctions are often used to treat nasal sinusitis and allergic rhinitis, and there are also reports of using Yuping Xanthium Powder to treat allergic rhinitis, etc., all of which illustrate the contribution of Xanthium in the treatment of rhinitis.

(2) Treatment of cancer Xanthium can be used as a compound preparation in different formulations, which has effectively cured many cases of nasopharyngeal cancer and nasal cavity cancer in clinical practice; treated 3 cases of penile cancer and cooperated with surgery for clinical cure; and treated intracranial tumors, Malignant tumors of the nervous system and brain tumors all have certain curative effects.

(3) Treatment of dental diseases It is reported in the literature that 98 cases of intractable toothache were treated with ancestral single prescription, and the curative effect was satisfactory. There are also reports that 15g each of Xanthium and Scrophulariae, decocted in water, is effective in treating toothache.

(4) Treatment of dermatitis Among the 38 patients with dermatitis, 25 cases were cured (65.8%), 10 cases were markedly effective (26.8%), and 3 cases were ineffective (7.9%), and the total effective rate was 92.1%.

(5) Treatment of upper respiratory tract infection It is reported in the literature that 30 outpatients with upper respiratory tract infection were treated with Xanthium cocklebur, 26 cases were cured (86.7%), 3 cases improved (10.0%), and 1 case was ineffective; The efficiency reaches 96.7%. Take the least 2 doses and the most 5 doses.

(6) Treatment of diarrhea Xanthium take about 50g ~ 70g, add 3000ml of water to soak for 30min, decoct with strong fire and then decoct with slow fire for 15min, filter out the medicinal liquid, cool to 35℃～38℃, soak in warm liquid or Soak the calf and foot 3 times a day; 46 cases were cured, the cure rate was 95.77%, 1 case was effective, the effective rate was 2.15%, and 1 case was ineffective (accounting for 2.15%).

(7) Treatment of otitis media It has been reported in the literature that cocklebur powder combined with antibacterial dry powder has achieved good therapeutic effects in the treatment of simple suppurative otitis media.

(8) Treat typhoid cocklebur in 6000g, immerse in water, heat and boil for 1 hour, filter out the decoction, decoct 3 times in the same way, mix the 3 decoctions, filter, simmer and concentrate to 2000ml, add paraben 10g , Shake well, effective treatment of 15 cases of typhoid fever, the fastest time for fever 10h, 7 cases of blood, feces, bile positive typhoid bacillus, all turned negative after treatment.

(9) Treatment of dysentery Take 100g of fresh cocklebur, wash and mash, add water to decoct for 15 minutes, remove slag, add 2 to 3 eggs, cook in the liquid medicine, take the liquid medicine and eggs together before the attack, 1 If it does not heal after the first time, it can be followed by the method and then taken again. It has a good curative effect on dysentery.

(10) Treatment of sores Cocklebur, burdock, raw rhubarb, etc., 1 dose a day, decocted in water and taken twice; 4-8 doses in a row, treating more than 30 cases of sores and furuncles, and all achieved good results.

(11) Other clinical applications There are also clinical reports on the use of cocklebur and its compound preparations in the treatment of low back pain, mumps, and lower extremity ulcers, all of which have received certain curative effects.

Fructus Xanthii is clinically used alone or combined with other traditional Chinese medicines to treat common and difficult diseases in rhinology such as acute and chronic rhinitis, sinusitis, and allergic rhinitis, and has achieved good curative effect. "Cangerzi San", "Canger Danggui Drink", "Cangxin Yuzhi Decoction", etc., are all famous prescriptions for clearing stagnation and heat, removing dampness and turbidity, and opening nose orifices. good. However, due to in-depth research on its chemical constituents and insufficient pharmacological screening, the active site and active ingredients are not clear and there is no feasible quality standard, which limits the further development and utilization of Xanthium.

(2) Research progress of sinusitis

Sinusitis is an acute or chronic inflammation of the sinuses caused by bacteria invading the sinuses and destroying the sinus tissue. The incidence of maxillary sinusitis is the highest in the site of disease, followed by ethmoid sinusitis, frontal sinusitis and sphenoid sinusitis. If all sinuses are involved, it is called pansinusitis.

Acute and chronic sinusitis is a kind of common disease and frequently-occurring disease, and according to bibliographical reports, only the United States just has 37 million chronic sinusitis patients; 140 million population), and the number of cases of sinusitis is increasing rapidly year by year. In China, due to different regions, the incidence of sinusitis fluctuates between 10.7% and 44.5%. According to clinical statistics, the incidence of chronic sinusitis accounts for 13.02% of newly diagnosed patients in ENT. Especially in southern China, due to the humid climate, the incidence of sinusitis is very high. More than one-third of the newly diagnosed patients in the Otolaryngology Department of Shanghai Changhai Hospital are patients with various types of sinusitis, and the patients are under 30 years old. 76.0%, 6.4% between 40 and 50 years old, 1% or 2% after 50 years old.

Due to factors such as narrow sinus ostia, unfavorable excretion, and allergic constitution, the disease often recurs and is difficult to heal. And this disease course of disease is long, is often accompanied by symptoms such as headache, dizziness, hypomnesis again, thereby brings extremely adverse effect to patient's study, work, life.

In addition, after sinusitis occurs, the inflammation can expand and invade adjacent tissues, which can easily cause osteomyelitis, orbital cellulitis, leptomeningitis, brain abscess, and severe suppurative sinusitis can even lead to sepsis, which directly endangers the patient's life. . Studies have shown that repeated stimulation of chronic sinusitis can also lead to cancer.

Modern studies have shown that chronic sinusitis is the result of a combination of many factors, but many of these factors have not yet been understood, and its pathogenesis has not been fully elucidated. Chinese and foreign scholars' researches on its pathogenesis mainly focus on the following aspects: ① bacterial or viral infection; ② ostiomeatal complex (OMC) obstruction; ③ mucociliary dysfunction;

The pathogenic bacteria of sinusitis are mostly pyogenic cocci, such as pneumococcus, hemolytic streptococcus, staphylococcus, etc., followed by bacilli, such as influenza bacillus, Proteus and so on. Clinically, the occurrence of sinusitis is mostly due to the mixed infection of two kinds of bacteria. Bacterial invasion damages the host's resistance, reduces the function of nasal mucosa cilia, and even paralyzes the defense mechanism of the mucosa.

Studies in recent years have shown that obstruction of the ostiomeatal complex caused by various reasons is the main cause of persistent sinusitis and recurrent attacks. At the same time, the above-mentioned links do not exist in isolation, but interact and interact with each other, resulting in persistent inflammatory reactions and irreversible pathological changes in the sinus mucosa.

The incidence rate of sinusitis is high, and the inflammation is easy to expand and cause serious complications, and it is easy to recur, and it is difficult to heal if it is protracted. At present, the commonly used western medicines in clinical practice are mainly antibiotics, decongestants, expectorants, analgesics, steroids and nasal sprays, etc., which are only suitable for patients with mild and early stages and patients before surgery, and most of them can only be cured to varying degrees. Improve symptoms, but can not completely cure sinusitis. About 2/3 of patients with sinusitis will have side effects of varying degrees after external use or taking these western medicines. For example, the most common is ephedrine nasal drops. Long-term repeated use can lead to nasal vasomotor dysfunction and slight degeneration of olfactory nerve endings. , and finally form drug-induced rhinitis; the abuse of antibiotics can easily lead to drug resistance of pathogenic bacteria, and may also destroy the balance of normal flora, make it imbalanced, and cause adverse consequences. To sum up, although there are many drugs for the treatment of sinusitis on the market, their clinical application is limited by their shortcomings such as unstable curative effect and varying degrees of side effects.

(3) Research progress of inflammation

Both exogenous damage factors and endogenous damage factors can cause various damage lesions of cells, at the same time, a series of complex reactions take place locally and throughout the body to limit and eliminate these damage factors, clear and Absorb necrotic tissue, cells, and repair damage, which is the body's defensive response, that is, inflammation (inflammation).

The local clinical features of inflammation are redness, swelling, pain and dysfunction, and the systemic reaction caused by inflammation includes fever and increased white blood cell count in peripheral blood. Inflammation is originally the body's first line of defense against viral infection, and it is the protection of the body. But when inflammation is severe or chronic, it has the potential to lead to heart attacks, colon cancer and Alzheimer's, among other diseases.

At present, anti-inflammatory and analgesic drugs commonly used in clinical practice are mainly non-steroidal anti-inflammatory drugs (abbreviation: NSAIDs). NSAIDs have been widely used for analgesia in patients with rheumatoid diseases and non-rheumatic diseases, including acute postoperative pain, chronic pain, biliary colic and renal colic and other abdominal pain and menstrual pain. Traditional non-steroidal anti-inflammatory drugs will inevitably cause damage to gastrointestinal tract, blood coagulation function and renal function due to the increase in dosage, course of treatment and age of patients, which restricts its application range.

Mechanism of action of NSAIDs: It inhibits the activity of cyclooxygenase (cyclooxyRenase) COX, thereby reducing the synthesis of prostaglandin PGs and achieving anti-inflammatory and analgesic effects. There are two isoenzymes of COX enzymes, basally expressed COX-1, whose main function is to maintain the normal physiological functions of gastrointestinal, kidney and other organs; inducible expressed COX-2, which can be found in the brain and local inflammation and play a role. The anti-inflammatory mechanism of NSAIDs is believed to be the inhibition of COX-2, while the occurrence of adverse reactions is due to the inhibition of COX-1.

Commonly used non-specific NSAIDs, such as ibuprofen, naproxen, and diclofenac sodium, have long and narrow structures and no selectivity, and can simultaneously interact with the active sites of COX-1 and COX-2. Combined, it can not only inhibit the activity of COX-1, but also inhibit the activity of COX-2. Therefore, it not only has anti-inflammatory and analgesic effects, but also often has adverse reactions such as gastrointestinal tract, blood coagulation function and renal function.

The main problem with NSAIDs treatment is adverse effects. As the age increases, the adverse reactions become more significant, including the gastrointestinal tract, kidneys, cardiovascular system, and central nervous system.

(4) Research progress of viral diseases and their treatment

According to statistics, 60% to 65% of epidemic infectious diseases are caused by virus infection. In August 2002, the seventh virology report presented by the World Congress of Virology held in Paris, France pointed out that there are 29 families, 7 subfamilies, 53 families, and more than 1,200 kinds of viruses that cause human diseases. There are 6 kinds of viral diseases with high incidence and great harm, such as acute and chronic hepatitis caused by hepatitis virus; cytomegalovirus (abbreviation: CMV) retinitis, interstitial pneumonia, herpetic Encephalitis, keratitis, genital herpes, herpes zoster, cold sores; bronchitis, pneumonia, measles, mumps, polio caused by respiratory virus infection; acute gastroenteritis and infantile diarrhea caused by enterovirus, etc. AIDS (abbreviation: AIDS) caused by human immunodeficiency virus (abbreviation: HIV) and so on.

Although humans have been constantly looking for effective weapons against viruses since the first day they knew about viruses, there is still a lack of specific drugs for the treatment of viral diseases. Clinically commonly used drugs include the following categories: ① Antiviral drugs that inhibit virus replication; ② Immunomodulatory drugs that enhance the immune function of the body; Anti-infective drugs for infection; ⑤ Vaccines to prevent virus infection; ⑥ Disinfectants to block virus transmission, etc.

(5) Research progress of analgesic drugs

Pain is a feeling shared by all human beings with great individual differences, and it is an alarm signal of damage to the body and internal organs. Pain is more difficult to study than other sensations because it is often intertwined with autonomic nervous activity, mental and emotional responses, and has great variability. For a long time, people's understanding of the etiology and pathology of pain, including migraine, rheumatoid arthritis, osteoarthritis, back pain, cancer pain and neuropathic pain, is still superficial. Yes, the treatment of neuropathic pain remains difficult. After more than 30 years of hard work, none of them have been satisfactorily resolved. The root cause is the slow progress in understanding the etiology and pharmacology of pain. Currently the most commonly used analgesics are non-steroidal anti-inflammatory drugs (abbreviated: NSAIDs) and opioids. There are 18 commonly used NSAIDs, and more than 100 are under research. Although their clinical curative effect is better, the adverse reactions are daunting, such as gastrointestinal reactions, gastric ulcer, gastric bleeding and allergic reactions of NSAIDs. Opioids are legally restricted due to their addictive properties. Especially for nociceptive pain (nociceptive pain), complex regional pain syndrome (complex regional pain syndrome, referred to as: CRPS) and neurogenic pain caused by trauma, inflammation or other diseases caused by nerve damage or disease is more difficult to treat. Big, strong narcotic pain relievers are also less effective. Fortunately, breakthroughs have been made in the understanding of the etiology and pharmacology of chronic pain in recent years, which has fundamentally changed the concept of pathology, physiology and therapeutics of chronic pain, and got rid of traditional NSAIDs and opioids. A batch of new analgesics have been developed based on the old framework of drugs. But most are in development or clinical trials. In addition, in recent years, Chinese and foreign scholars have targeted N-type voltage-sensitive calcium ion channels, nerve growth factor receptors, adenosine kinases, substance P receptors, and cannabinol receptors, and have also developed therapeutic drugs for neuropathic pain. A lot of work has been done, and the search for analgesics for the treatment of neurogenic pain has become a frontier topic in clinical and basic and therapeutic drug research.

(6) Research progress on pharmacological activity of phenolic acids

1. Anti-platelet aggregation and prevention of thrombosis The research results of intravenous injection of seven water-soluble components of Salvia miltiorrhiza on rat platelet aggregation showed that salvianolic acid A and salvianolic acid B have a strong effect on collagen-induced platelet aggregation. Inhibitory effect, the IC ₅₀ of which is 0.32mmol/L and 0.26mmol/L, respectively, for ADP-induced aggregation, salvianolic acid A is effective but salvianolic acid B is ineffective. Rosmarinic acid has a mild effect of preventing thrombosis, and it is one of the active ingredients of Danshen antithrombotic effect. Rosmarinic acid (iv) 50mg/kg, 100mg/kg inhibited venous thrombosis in rats by 41% and 55.8% (P<0.05).

2. Improve microcirculatory disturbance By observing the changes in the caliber, blood velocity and flow rate of the mesenteric arterioles in mice, it was shown that the topical administration of salvianolic acid A and salvianolic acid B has a positive effect on the mesentery of mice induced by adrenaline. The microcirculation disorder can be significantly improved, and the intravenous injection of salvianolic acid A can also have the same effect.

3. The anti-lipid peroxidation salvianolic acid A, salvianolic acid B, salvianolic acid C and rosmarinic acid have stronger inhibitory effects on the generation of malondialdehyde (MDA) in mouse liver homogenate than vitamin E. The order of inhibition is salvianolic acid A > salvianolic acid C > salvianolic acid B > rosmarinic acid, and the scavenging effect of the four components on hydroxyl radicals is 10-100 times stronger than that of mannitol. For mice with acute alcoholism caused by oral administration of 50% ethanol 15ml/kg, after treatment with rosmarinic acid (100mg/kg×3, po), the MDA content in the liver of the mice was significantly reduced. Effects of salvianolic acid A, salvianolic acid B and rosmarinic acid on rat brain and liver induced by vitamin C-nicotinamide adenine dinucleotide phosphate (NADPH) or by Fe ²⁺ -cysteine , kidney microsomes have a strong inhibitory effect on lipid peroxidation, and their effects are followed by salvianolic acid A, salvianolic acid B, and rosmarinic acid, which are 100% stronger than the antioxidant vitamin E 1000 times. Salvianolic acid A can inhibit the loss of mitochondrial lipid peroxidation and ATPase activity in rat heart and liver induced by Fe ²⁺ -cysteine, and inhibit the mitochondrial swelling caused by lipid peroxidation and the decrease of liver mitochondrial membrane fluidity .

4. Preventive effect on cardiotoxicity of doxorubicin The cardiotoxicity of doxorubicin is caused by the generation of free radicals in the heart and damage to myocardial mitochondria. Salvianolic acid A can scavenge OH· produced by doxorubicin, and has a preventive effect on cardiotoxicity caused by doxorubicin, but has no effect on the antitumor effect of doxorubicin.

5. Anti-gastric ulcer Salvianolic acid A can inhibit pig stomach H ⁺ -K ⁺ /ATPase and PNPP enzyme, with IC ₅₀ of 5.2×10 ^-7 and 1.7×10 ^-6 mol/L, respectively. Salvianolic acid A (25mg/kg, ip) has a strong inhibitory effect on gastric acid secretion in rats with pylorus ligation, and the same dose of salvianolic acid A can weaken the gastric injury caused by water immersion or forced stress. These results indicated that salvianolic acid A has gastric acid secretion inhibitory and antiulcer activity by inhibiting gastric H ⁺ -K ⁺ /ATPase.

6. Relieve symptoms of uremia The magnesium salt of salvianolic acid B can reduce the concentration of urine nitrogen, creatinine, methylamidine guanidine, and guanidinosuccinic acid in the blood of mice with uremia caused by taking adenine, thereby significantly reducing the symptoms of uremia. Rat uremic symptoms.

7. Inhibition of adenylate cyclase Rosmarinic acid, shikonic acid and their methyl esters have effects on the basic adenylate cyclase of rat brain and the adenylate cyclase activated by 5umol/L diterpene derivatives Strong inhibitory effect.

8. Others For example, ginsenosides and some phenolic acid compounds are the main anti-aging active ingredients of ginseng; tea polyphenols are the general term for a variety of phenolic derivatives, accounting for about 25%, which include catechins, anthocyanins, flavonoids and flavonols , phenolic acid four compounds. They contain polyhydric groups, have the properties of phenols, and have significant pharmacological effects, such as lowering blood sugar, blood lipids, and preventing atherosclerosis; they can interact with bacteria such as E. Combining, making it precipitate, inhibiting the growth of bacteria, and promoting the loss of activity of various bacteria; it can also produce toxic substances such as nicotine, ahydromorphine, and cinchona alkali in the human body and heavy metals such as lead, cobalt, silver, and copper that are harmful to the human body. Precipitate, weaken or eliminate hazards.

According to literature search, so far, no report on tracking and screening the active part or active ingredient of Fructus Xanthium has been found, nor has there been any relevant report on the use of the active part extract of Fructus Xanthium.

Contents of the invention

The technical problem to be solved by the present invention is to disclose a new application of the total extract of the traditional Chinese medicine Xanthium Fructus, so as to overcome the above-mentioned defects in the prior art.

That is to say, the present invention intends to clarify the active parts and active ingredients of the Chinese medicine Xanthium and its preparation method, and then use the total extract of Xanthium to prepare anti-inflammatory reaction products; that is, the present invention relates to the traditional Chinese medicine Xanthium The total glycoside extract and its preparation method, and the application of the total glycoside extract and its composition, such as pharmaceutical composition, etc. in the preparation of anti-inflammatory reaction products.

The anti-inflammatory response products refer to products for anti-inflammatory and related diseases, products for anti-virus and related diseases, products for analgesic and related diseases, products for diagnosis, detection, treatment or research of acute and chronic rhinitis and related diseases Products, or products for the diagnosis, detection, treatment or research of sinusitis and related diseases; wherein, the anti-inflammatory response products include anti-inflammatory response drugs, anti-inflammatory response reagents, anti-inflammatory response food or anti-inflammatory response One or more of inflammatory response beverages, etc., preferably anti-inflammatory response drugs.

(1) Technical conception

Independently developing innovative drugs is an urgent task for China at present. Chinese medicine has a long history, and has accumulated rich experience in the prevention and treatment of diseases with Chinese herbal medicine. Therefore, finding effective active ingredients from Chinese medicine is an effective way, and it is also the advantage of China's innovative drug development.

The inventor retrieved books such as "Newly Edited National Chinese Patent Medicine", "Handbook of Commonly Used Clinical Prescriptions", "Encyclopedia of Chinese Commonly Used Chinese Proprietary Medicines" and other books, and conducted a survey and statistical analysis of hospital medications and commercially available medicines for patients with ENT patients, and found that in the treatment of Among the 30 commonly used traditional Chinese medicine prescriptions for rhinitis and sinusitis, 23 kinds contain cocklebur, accounting for 76.7%. Therefore, it is speculated that there may be substances or substance groups with anti-inflammatory, antibacterial, and antiviral activities in Fructus Xanthii. However, due to in-depth research on its chemical constituents and insufficient pharmacological screening, the active site and active ingredients are not clear and there is no feasible quality standard, which limits the further development and utilization of Xanthium.

Therefore, the present invention conducts systematic research on the chemical constituents of the total extract of Xanthium solitary, and screens and proves the activity and use of the total extract.

Through chemical composition research, it is proved that the main components in the active part of Xanthium Fructus are diterpene glycosides and phenolic acids. According to the literature search, the inventors found that most of the phenolic acid compounds have a variety of significant pharmacological activities, such as the famous salvianolic acid, ginkgolic acid, etc. The analgesic, antiviral activity and the clinical efficacy of treating a variety of nasal diseases should be mainly exerted by the efficacy of the active part of Xanthia glycosides, and the research results have also proved and confirmed that Xanthia glycosides extract has Significant pharmacological activity.

(2) Xanthium total glycosides extract

1. Composition of total glycosides extract of Xanthium fructus

The inventors have found that Fructus Xanthii mainly contains a large amount of volatile oil, sesquiterpene lactones and water-soluble glycosides through systematic chemical composition separation, purification and structural identification of experimental research on Fructus Xanthii medicinal material.

The components in the extract of total glycosides of Xanthium mainly include two types of compounds such as diterpene glycosides and phenolic acids; the above two types of compounds are the main active ingredients in the extract of total glycosides of Xanthium, and also include other ingredients.

Wherein, the content of diterpene glycosides in the total glycosides extract ranges from 0.5% to 99% (W/W, that is, weight percentage), preferably 5.0% to 95.0%;

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

Wherein, the content range of other components in the total glycoside extract is less than 0.5%, and other components include one or more of tannins, monosaccharides or polysaccharides, xanthones or carotenoids, etc. .

Existing studies have shown that other components have at least no negative impact on the efficacy or use of the drug, and the cost of isolating these components is relatively high, so there is currently no need for further purification.

2. The general chemical structure formula of the diterpene glycosides in Fructus Xanthii is as follows:

Wherein, R ₁ is one of hydrogen, methyl or carboxyl; R ₂ and R ₃ are one of hydrogen, sulfonic acid, acetyl, propionyl or dimethylpropionyl.

Table 1 lists the corresponding relationship between R ₁ , R ₂ , and R ₃ groups in the structural formulas of some diterpene glycosides:

When R ₁ =COOH, R ₂ =R ₃ =H, it means diterpene glycoside compound A;

When R ₁ =COOH, R ₂ =H, R ₃ =COCH ₂ CH(CH ₃ ) ₂ , it means diterpene glycoside compound B;

When R ₁ ＝COOH, R ₂ ＝SO ₃ ^- , R ₃ ＝COCH ₂ CH(CH ₃ ) ₂ , it means diterpene glycoside compound C;

When R ₁ ＝H, R ₂ ＝H, R ₃ ＝COCH ₂ CH(CH ₃ ) ₂ , it means diterpene glycoside compound D;

When R ₁ =H, R ₂ =SO ₃ ^- , R ₃ =COCH ₂ CH(CH ₃ ) ₂ , it means diterpene glycoside compound E.

Table 1. Chemical structural formulas of several diterpene glycosides in Fructus Xanthii

R ₁ R ₂ R ₃ A COOH H h B COOH H -COCH ₂ CH(CH ₃ ) ₂ C COOH SO ₃ ^- -COCH ₂ CH(CH ₃ ) ₂ D H H -COCH ₂ CH(CH ₃ ) ₂ E H SO ₃ ^- -COCH ₂ CH(CH ₃ ) ₂

3. The general chemical structure formula of the phenolic acid compounds in Fructus Xanthii is as follows:

Wherein, R ₁ is one of hydrogen or an alkali metal element; R ₂ , R ₃ and R ₄ are one of hydrogen or caffeoyl. Caffeoyl is Caffeoyl, the structure is as follows:

Table 2 lists the corresponding relationship with R ₁ , R ₂ , and R ₃ groups in the structural formulas of some phenolic acid compounds.

When R ₁ ＝H, R ₂ ＝R ₃ ＝R ₄ ＝Caffeoyl, it means phenolic acid compound A;

When R ₁ =R ₃ =H, R ₂ =R ₄ =Caffeoyl, it means phenolic acid compound B;

When R ₁ =R ₂ =H, R ₃ =R ₄ =Caffeoyl, it means phenolic acid compound C;

When R ₁ =K, R ₂ =R ₄ =H, R ₃ =Caffeoyl, it means phenolic acid compound D.

Table 2. Chemical structural formulas of several phenolic compounds in Fructus Xanthii

R ₁ R ₂ R ₃ R ₄ A H Caffeoyl Caffeoyl Caffeoyl B H Caffeoyl H Caffeoyl C H H Caffeoyl Caffeoyl D K H Caffeoyl H

(3) The preparation method of total glycosides extract of Fructus Xanthii

The preparation method of the said total glycosides extract of Xanthium Fructus in the present invention comprises the following steps:

(1) Extraction: get some raw materials of Fructus Xanthium, extract, and obtain the extract, i.e. the total extract of Fructus Xanthium;

(2) Separation and purification: Separation and purification of the extract to obtain the total glycosides extract of Xanthium fructus.

The extraction method includes all available 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 to say, it includes the commonly used ultrasonic extraction method, immersion extraction method, percolation extraction method, decoction extraction method, heating reflux extraction method or continuous reflux extraction method. One or more of extraction methods, etc.; the number of extractions may be one or multiple times. At the same time, factors such as the crushing degree of raw materials, extraction time, extraction temperature, and equipment conditions can also affect the extraction efficiency and must be considered. For all technical materials including technical details of various extraction methods, you can refer to relevant teaching materials and relevant technical documents.

The extraction solvent used in the solvent extraction method described above is a conventional extraction solvent in the art, that is to say, it includes a kind of three types of extraction solvents such as common water reagents, hydrophilic organic solvents or lipophilic organic solvents. or more;

The water reagent is one of water, acid water or alkaline water, etc.;

Described hydrophilic organic solvent is the so-called water-miscible organic solvent, including one or more in ethanol, ethanol aqueous solution or methanol, etc., preferably ethanol or ethanol aqueous solution of different concentrations, etc. One or more, more preferably aqueous ethanol solutions of different concentrations;

The lipophilic organic solvent is a so-called water-immiscible organic solvent, including one or more of sherwood oil, chloroform, ether, ethyl acetate, methylene dichloride or ethylene dichloride, etc. , preferably one or more of petroleum ether, chloroform, diethyl ether, ethyl acetate, dichloromethane or dichloroethane, etc., more preferably one or more of chloroform, diethyl ether, ethyl acetate or dichloromethane, etc. kind.

The total extract of Fructus Xanthium obtained by the above extraction method needs to be further separated and purified.

The separation and purification method is all available methods known in the art, including solvent separation, solvent extraction (including extraction, countercurrent continuous extraction, countercurrent distribution or droplet countercurrent distribution, etc. or Multiple), macroporous adsorption resin method, precipitation method, salting-out method, column chromatography or crystallization and recrystallization and fractional crystallization etc. one or more; These methods are all known techniques in the art, When further confirmation is required, it is easy to consult all technical information including technical details from relevant textbooks and relevant technical documents.

In order to make the product stable, easy to use and store, and improve the bioavailability of the product, an optimization step can also be added, namely step 3:

(3) dry.

The drying method is a method known in the art that can be used, including normal pressure drying methods such as drying in an oven, vacuum drying, vacuum drying, spray drying, freeze drying, far-infrared heating and drying methods or microwave drying. One or more of drying methods, etc.

In the specific implementation, it is necessary to adopt the appropriate method and select appropriate conditions and other necessary measures according to the existing funds, technology and related requirements to achieve the expected goal. And these methods are all well-known technologies in the art. When further confirmation is needed, it is easy to consult all technical materials including technical details from relevant teaching materials and relevant technical documents.

(4) Uses of Xanthium glucosides extract

1 Overview

A further object of the present invention is to provide a new application of the total glycosides extract of Xanthia Fructus in the preparation of anti-inflammatory reaction products, and the products include one or more of drugs, reagents, food or beverages, etc., preferably drugs.

Through the screening of pharmacological activity, it is proved that the total glycosides extract of Xanthium fructus obtained from the extraction part of n-butanol or the elution part of macroporous adsorption resin with 5-75% ethanol is anti-inflammatory, analgesic, anti-viral or diagnosis, detection and treatment. and active sites for research on rhinopathy and related conditions. The completed acute toxicity experiment proves that the maximum tolerated dose of the active part by intragastric administration in mice exceeds 2.0g/kg, which is equivalent to 440 times the clinically recommended drug dose, indicating that the effective part is safe and reliable, and solves the problem of compound Chinese medicine. The problem of complex ingredients, low content of active ingredients and toxic ingredients.

The inventors found that the total glycosides extract of Xanthium fructus has anti-inflammatory and analgesic effects, can significantly reduce inflammation and swelling, inhibit capillary permeability, and has antiviral effects.

At the same time, the inventors found that the total glycosides extract of Xanthium can reduce the infiltration of nasal mucositis cells caused by rhinitis and sinusitis, eliminate nasal mucosal edema, inhibit submucosal connective tissue hyperplasia, thereby improving nasal mucosal swelling and ventilation function, and reducing nasal congestion. secretion, so that the inflammation subsides, the effect is better than that of biyuanshu oral liquid, which is commonly used in clinical practice.

2. The use method and requirements of the total glycosides extract and composition of Xanthium fructus

The total glycosides extract of Xanthia fructus of the present invention can be used alone or in combination with other active components, including products for the diagnosis, detection, treatment or research of related diseases, including drugs, reagents, food or beverages, etc., especially drug.

In terms of specific use, the total glycosides extract of Xanthium fructus in the present invention can be used alone, and can also be used together with many other chemical substances. Regardless of whether these chemical substances have biological activity or have the function of treating diseases, including auxiliary functions such as synergistic amplification, antagonizing or alleviating the side effects of Xanthia glycosides extract, etc., these chemical substances include pharmaceutically acceptable carriers, One or more of foods, natural products, chemically synthesized drugs or human medicines; preferably one or more of pharmaceutically acceptable carriers or foods; more preferably pharmaceutically acceptable carriers.

As used herein, "pharmaceutically acceptable carrier" includes any and all physiologically acceptable solvents, dispersion media, coatings, 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, etc., and combinations thereof. In many cases it will be desirable to include in the compositions isotonic agents, for example, one or more of a sugar, a polyol such as mannitol, sorbitol, sorbitol, sodium chloride, and the like. The pharmaceutically acceptable carrier can also contain a small amount of auxiliary substances, such as one or more of wetting agents or emulsifiers, preservatives or buffers, etc., which enhance the effective period or potency.

In terms of specific classification, said pharmaceutically acceptable carrier refers to a conventional drug carrier in the field of medicine, including excipients, such as one or more of starch or water; lubricants, such as glycerin or magnesium stearate, etc.; disintegrants, such as microcrystalline cellulose, etc.; fillers, such as one or more of starch or lactose; binders, such as pregelatinized starch One or more of dextrin, cellulose derivatives, alginate, gelatin or polyvinylpyrrolidone, etc.; osmotic pressure regulator, such as one or more of glucose, sucrose, sorbitol or mannitol, etc. ; diluents, such as water, etc.; disintegrants, such as one or more of agar, calcium carbonate or sodium bicarbonate, etc.; absorption enhancers, such as quaternary ammonium compounds, etc.; surfactants, such as cetyl alcohol, etc.; Adsorption carrier, such as one or more in kaolin or bentonite; Lubricants, such as one or more in talcum powder, calcium stearate, magnesium stearate or polyethylene glycol; In addition, also Other adjuvants, such as one or more flavoring agents or sweetening agents, can be added to the composition.

For example, dissolving, suspending or emulsifying the active ingredient total glycosides of Xanthia fructus in a suitable aqueous solvent (for example, one or more of distilled water, normal saline or Green's solution, etc.) or in an oily solvent (for example , vegetable oil such as one or more of olive oil, sesame oil, cottonseed oil, corn oil or propylene glycol, etc.), the injection preparation can be prepared, wherein the solvent may contain a dispersant (for example, polysorbate 80, polyoxygen One or more of ethylene hardened castor oil 60, polyethylene glycol, benzyl alcohol, chlorobutanol or phenol, etc.), osmotic pressure regulator (for example, sodium chloride, glycerin, D9-mannose, D- one or more of sorbitol or glucose, etc.). In this case, if necessary, additives such as solubilizers (for example, one or more of sodium salicylate or sodium acetate, etc.), stabilizers (for example, human serum albumin, etc.), pain relievers Agents (for example, benzyl alcohol, etc.) and the like.

The total glycosides extract of Xanthium fructus mentioned in the present invention can also be used in combination in the form of a composition, especially with other chemical substances such as drugs to treat animals, especially mammals, including humans or other animals. is a similar composition. The mammals include one or more of humans, mice, rats, sheep, monkeys, cows, pigs, horses, rabbits, dogs, chimpanzees, baboons, marmosets, rhesus monkeys or rhesus monkeys. For example, the extract of total glucosides of Xanthia fructus of the present invention can be added into a pharmaceutical composition suitable for administration to a subject. Generally, the pharmaceutical composition comprises the extract of total glycosides of Xanthogside of the present invention and a pharmaceutically acceptable carrier.

The composition, particularly the pharmaceutical composition, of the total glycosides of Xanthia can be in various forms, including one or more of dosage forms such as liquid, semi-solid and solid; wherein said pharmaceutical composition includes therapeutically effective A certain amount of total glycosides extract of Xanthia fructus is an active ingredient, and one or more pharmaceutically acceptable carriers.

The pharmaceutical composition of the total glycosides extract of Xanthia fructus can be prepared into various dosage forms by conventional production methods known in the art, for example, the active ingredient is mixed with one or more carriers, and then prepared into the desired dosage form. Described dosage form comprises one or more in tablet, capsule, granule, suspension, emulsion, solution, syrup or injection etc., takes oral or injection (comprising intravenous injection, intravenous drip, intramuscular injection) One or more of injection or subcutaneous injection, etc.), mucosal dialysis, etc., for anti-inflammatory, antiviral, analgesic, and acute and chronic rhinitis, sinusitis and related disease diagnosis, detection, treatment or scientific research.

The pharmaceutical composition preferably contains 0.5% to 99% by weight of the active ingredient Xanthia glycosides extract, more preferably contains 1% to 95% by weight of the active ingredient Xanthia glycosides extract, and most preferably contains The ratio is 5% to 90% of the active ingredient total glycosides extract of Xanthium fructus.

The pharmaceutical composition of the total glycosides extract of Xanthia fructus generally must be sterile and stable under production and storage conditions. The composition can be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable to high drug concentration. The sterile injectable solution is prepared by adding the required amount of the total glycosides extract of Xanthia officinalis together with one or a combination of the above-mentioned ingredients as required into an appropriate solvent, followed by sterile filtration. In general, dispersions are prepared by adding the total glycosides extract of Xanthia fructus to a sterile vehicle containing a basic dispersion medium and the required other ingredients as 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 solutions can be maintained, for example, by coatings such as lecithin, by maintaining the desired particle size in the case of dispersions and by using surfactants. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example a salt of monostearate or gelatin.

When used for patients, the dose of the total glycosides extract of Xanthia fructus of the present invention is 5-20 mg/kg·d, and the dose or dosage is usually determined according to the age and weight of the patient or user, as well as the physical condition or the condition of the patient's symptoms. Decide.

The extract of total glycosides of Xanthia fructus of the present invention and the pharmaceutical composition thereof may include the extract of total glycosides of Xanthium fructus of the present invention in a "therapeutic effective amount" or "preventive effective amount". "Therapeutically effective amount" refers to an amount effective to achieve the desired therapeutic effect at dosages and for periods of time necessary. The therapeutically effective amount of Xanthia glucosides extract can vary depending on factors such as the individual's condition, age, sex, and weight, and the ability of the Xanthia glucosides extract to elicit a desired response in the individual. A therapeutically effective amount also refers to an amount in which the beneficial therapeutic effects of the total glycosides extract of Xanthia officinalis outweigh any toxic or harmful effects thereof. "Prophylactically effective amount" refers to the amount effective to achieve the desired prophylactic effect at the necessary dose and time. Because prophylactic doses are used in subjects who are pre-ill or early in the course of the disease, the prophylactically effective amount is usually less than the therapeutically effective amount. The typical non-limiting range of the therapeutic or preventive effective dose of the total glycosides extract of Xanthia fructus of the present invention is 5-20 mg/kg, more preferably 5-10 mg/kg. It should be noted that the dose value will vary according to the type and severity of the disease to be alleviated, that is to say, when used in a patient, the dose or amount of the total glycosides extract of Xanthia fructus according to the present invention is generally based on the age of the patient or user and weight and physical condition or condition of the patient's symptoms. In addition, it is understood that for any particular subject, the particular dosage regime should be adjusted over time according to the individual needs and the professional judgment of the person administering or supervising the administration of the composition, and that the dosage ranges set forth herein are illustrative only intended to limit the scope or practice of the claimed compositions.

That is to say, it is necessary to change the dosage or usage amount of the total glycosides of Xanthia fruticosa extract of the present invention each time and/or daily according to the object to be treated, the route of administration, the disease and condition to be treated, etc. For example, for intravenous administration to mammals, especially adults (eg body weight 60kg), the single dose of the total glycosides extract of Xanthia fructus is about 5-10 mg, preferably about 10 mg, preferably 1-3 times a day. Dosage unit adjustments may be made to optimize the desired response (eg, a therapeutic or prophylactic response). For example, a single bolus may be administered, several averaged amounts may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially 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 suitable as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of an active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Xanthium total glycosides extract. The specifications of the dosage unit form of the present invention are determined by and directly depend on (a) the unique characteristics of the Xanthia glucosides extract and the specific therapeutic or preventive effect to be achieved, and (b) when mixing such Intrinsic limitations in the technology of total glycosides extract of Xanthia fructus for the treatment of individual sensitivity.

3. The pharmaceutical dosage form and administration route of the total glycosides extract of Xanthium fructus and its composition

The total glycosides extract of Xanthia fructus and its composition in the present invention prepare anti-inflammatory reaction products, wherein the products prepared according to the requirements in the field of beverage and food technology can be anti-inflammatory, anti-viral, analgesic, and diagnosis, detection, and treatment Or research acute and chronic rhinitis, sinusitis and related diseases; products prepared according to the requirements of the medical technology field can be used for the treatment or health care of patients. Substances are mixed or combined for direct or indirect use in the preparation of medicines for treatment or health care. The chemicals described here are the same as those described earlier in this section.

In the present invention, the required materials include the raw materials of the present invention, the above-mentioned supporting chemical substances, etc., which should be food-grade or pharmaceutical-grade materials according to actual conditions and needs.

The total glycosides extract of Xanthia fructus and its composition described in the present invention can be administered by various methods known in the art, although the recommended route of administration/administration in many therapeutic uses is spray or oral administration medication. However, the skilled artisan will appreciate that the route/mode of administration will vary with the desired result. In certain implementations, the active compound can be prepared with a carrier that will protect the compound from rapid release, such as an air release formulation, including one of an implant delivery system, a transdermal patch delivery system, or a microencapsulated delivery system, or the like. Various. In addition, biodegradable, biocompatible polymers, such as one or more of ethylene ethyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, or polylactic acid, can also be used. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art (see, e.g., Sustained and Controlled Release Drug Delivery Systems, edited by J.R. Robinson, Marcel Dekker, Inc., New York, 1978).

The total glycosides extract of Xanthia fructus and its composition described in the present invention are usually administered to patients in need of such treatment through one or more methods of oral administration, nasal inhalation, rectal or parenteral administration and the like.

For oral administration, it can be made into one or more of conventional solid preparations such as tablets, powders, granules or capsules. During implementation, the total glycosides extract of Xanthii of the present invention can be taken orally together with, for example, an inert diluent or an assimilable edible carrier. The total glycosides extract of Xanthia fructus (and other components, if necessary) can also be enclosed in hard or soft shell gelatin capsules, compressed into tablets or directly added to the diet of the subject. For oral therapeutic administration, the total glycosides extract of Xanthii can be added together with excipients and formulated as edible tablets, buccal tablets, tinctures, capsules, suspensions, syrups or wafers, etc. One or more of the forms used.

In order to administer the xanthia glycosides extract of the present invention in addition to parenteral administration, it may be necessary to coat the xanthia glycosides extract with a material that prevents its inactivation or combine it with the xanthia glycosides extract. give together. Supplementary active compounds can also be incorporated into the compositions. In a specific implementation, the total glycosides extract of Xanthia fructus of the present invention is co-formulated and/or co-administered with one or more other therapeutic drugs that can be used for treating diseases. This combination allows advantageous use of lower doses of the administered therapeutic agent, thus avoiding possible toxicity or complications associated with various monotherapies.

Made into one or more of liquid preparations such as water, oil suspension or other liquid preparations, such as one or more of syrup or elixir; for parenteral administration, it can be made into injection One or more of the solutions, aqueous or oily suspensions used.

Among the above-mentioned use forms, the preferred form is one or more of tablets, coated tablets, capsules, suppositories, nasal sprays, nasal drops or injections, etc., more preferably nasal sprays, nasal drops One or more of formulations, tablets, etc., particularly preferably nasal sprays.

In addition, the medicinal materials used in the extract of total glycosides of Xanthia fructus can also be directly used to prepare anti-inflammatory reaction products alone in some cases, and can also be mixed or combined with many chemical substances, directly or in the form of a composition Indirectly used in the preparation of anti-inflammatory reaction products. The chemicals described here are the same as those described earlier in this section.

For example, the powder of medicinal materials used in the extract of total glucosides of Xanthium is used to prepare various dosage forms of anti-inflammatory products, especially drugs, or the powder of raw materials used in extracts of total glycosides of Xanthia is related to The excipients are used to prepare various dosage forms of anti-inflammatory reaction products, especially drugs, or the powder of medicinal raw materials used in the extract of total glucosides of Xanthia fructus is used to prepare anti-inflammatory reaction products such as drugs. Inflammatory reaction products such as various dosage forms of drugs, or the powder of raw materials used in the extract of cocklebur glucosides and related auxiliary drugs are used to prepare various dosage forms of anti-inflammatory reaction products such as drugs, such as tablets One or more of formulations, capsules, granules or suspensions, etc., preferably capsules.

One of the methods is to fill the powder of the raw materials used in the extract of total glucosides of Xanthia Fructus into capsules, and the second method is to mix the powder of raw materials used in the extract of total glucosides of Xanthia Fructus with related Preparation of anti-inflammatory response products such as medicines are filled together as capsules. The third method is to fill the powder of the raw materials used for the total glucosides of Xanthia glycosides together with related auxiliary drugs into capsules; the fourth method is The powder of the medicinal raw materials used in the extract of total glucosides of Xanthia glycosides and related auxiliary materials are directly pressed into tablets or granules according to conventional methods. The fifth method is to extract the raw materials of medicinal materials used in the Powder, related preparation of anti-inflammatory reaction products such as drugs and related excipients are directly compressed into tablets or granules according to conventional methods. Auxiliary drugs and related auxiliary materials are directly compressed into tablets or granules according to conventional methods.

In addition to the above-mentioned six basic methods, it is also possible to select other forms of medicinal materials used in the extract of total glucosides of Xanthium or to process the raw materials used in the extraction of total glucosides of Xanthia by methods known in the art, Products such as medicines containing medicinal materials used in the extract of total glucosides of Xanthia fructus are prepared in various dosage forms. However, it should be noted that when directly using the medicinal materials used in the extract of cocklebur glycosides, the required dose of cocklebur glycosides should be calculated according to the dosage requirements of the extract of cocklebur glycosides used. The amount of medicinal raw materials used in the total glycosides extract of Auricularia chinensis.

In summary, the total glycosides extract of Xanthia fructus and its composition of the present invention can be used to prepare anti-inflammatory products, preferably medicines and foods, and more preferably medicines.

(5) Technical expertise

The present invention studies the total glycosides extract of Xanthium Fructus pertinently. The raw material is safe to use, takes into account each other, has multiple functions, and exerts its effect to the maximum extent, and has a particularly wide range of use, so it is easy to popularize and apply, and can be used in a relatively short period of time. Generate huge social and economic benefits within a short period of time.

That is to say, the research results of the present invention can be further developed into anti-rhinitis, sinusitis, anti-inflammatory and analgesic and anti-viral products, especially new drugs, which have clinical application prospects. The product has the advantages of definite curative effect, low effective dose, and no side effects, and has significant advantages compared with similar products in China and abroad. Acute and chronic sinusitis are clinically common and frequently-occurring diseases. With the increasing global air pollution and the deterioration of the environment, it is expected that the incidence of sinusitis and other respiratory diseases will continue to rise. Therefore, the present invention can also lay a good foundation for subsequent new drug research, and further generate greater economic and social benefits.

After long-term pharmacological tests, the total glycosides extract of Xanthium Fructus has anti-inflammatory, anti-viral, analgesic, and the activities of treating acute and chronic rhinitis, sinusitis and related diseases. Through the metabolism of gastrointestinal bacteria, the effect is more direct, and the differences in the activity of gastrointestinal flora between individuals are avoided, resulting in differences in metabolites and differences in drug efficacy, thereby overcoming the problem that the dosage of traditional Chinese medicine is difficult to control. , and increased bioavailability.

The total glycosides extract of cocklebur of the invention has strong pharmacological action, rich sources of raw materials, simple preparation process and high yield.

The total glycosides extract of Xanthium is also an active form in the biotransformation of the gastrointestinal tract, and the direct use of the total glycosides extract of Xanthium has high bioavailability of the drug, and the dosage can be accurately controlled. The properties of the total glycosides extract of Xanthium Fructus are stable, and the quality of the preparation prepared by using the total glycosides of Xanthium Fructus extract is stable.

In addition, the total glycosides extract of Xanthia Fructus is stable in chemical properties, and has obvious effects of anti-inflammation, anti-virus, analgesia, and treatment of acute and chronic rhinitis, sinusitis and related diseases, so it is more suitable for anti-inflammatory, anti-virus, analgesic, etc. Pain and the industrial production of drugs for the treatment of acute and chronic rhinitis, sinusitis and related conditions.

In a word, the present invention actively adapts to the needs of work in the fields of modern medical treatment, food, beverage and scientific research and the needs of humanized services, and is a safe raw material for preparing anti-inflammatory reaction products, which can directly or indirectly anti-inflammatory, anti-virus, sedative pain and the diagnosis, detection, treatment or research of acute and chronic rhinitis, sinusitis and related conditions.

Detailed ways

The present invention studies the application of the total glycosides extract of Xanthia fructus, and provides a raw material that can be used to prepare anti-inflammatory reaction products such as medicines, etc. or anti-inflammatory reaction products, especially medicines, which is convenient for the medical industry and related industries such as Safe use in food, beverage and other fields, including disease diagnosis, detection, treatment or scientific research.

Taking the solvent extraction method as an example, the specific operation method for preparing the total glycosides extract of Xanthium fructus is described in detail below.

(1) Extraction: take some Xanthium medicinal materials, add an extraction solvent to mix, extract, filter and concentrate the extract, and obtain the total extract of Xanthium medicinal materials.

(2) Separation and purification: the total extract of Xanthium medicinal materials is subjected to separation and purification steps to obtain total glycosides extract of Xanthium.

The extraction, separation and purification methods mentioned here include solvent extraction, solvent extraction or macroporous adsorption resin method, column chromatography and other extraction, separation and purification methods known in the art as mentioned above. Specifically:

In step one:

① In the solvent extraction method adopted, the extraction solvent is one or more of water, alcohols or other organic solvents, and the alcohol extraction solvent includes methanol, ethanol, ethanol aqueous solution, propanol or butanol One or more in alcohol etc., described other organic solvents include ethyl acetate, acetone etc., preferably organic solvents such as water, methanol, ethanol, ethanol aqueous solution, propanol, butanol, ethyl acetate, acetone or its One or more of the mixed solvents, more preferably one of ethanol or ethanol aqueous solution, especially preferably 60%-90% ethanol aqueous solution.

② In the solvent extraction method adopted, the extraction method is preferably one or more of ultrasonic extraction, percolation extraction or heating reflux extraction, and further preferably heating reflux extraction; the number of extractions can be one or more times.

③Filter the extract under reduced pressure and concentrate to obtain the total extract of Xanthium medicinal material. The yield of the total extract of Xanthium medicinal material is 5.0% to 20%, that is, the total extract of Xanthium medicinal material accounts for the total weight of Xanthium medicinal material 5.0% to 20% of that.

In step two:

① The separation and purification method adopted is preferably solvent extraction method, that is, the total extract of cocklebur medicinal material is dispersed in water, degreased with petroleum ether, and then extracted with an appropriate organic solvent to remove fat-soluble components to obtain the extraction residue; or directly use An appropriate organic solvent is extracted to remove the fat-soluble components to obtain an extraction residue; the appropriate organic solvent can be extracted by one or more of chloroform, dichloromethane, ether or ethyl acetate, etc.; the number of extractions Can be one or more times.

②In the solvent extraction method adopted, it is also necessary to continue to extract the extraction residue obtained after extracting the fat-soluble components with n-butanol, separate the n-butanol layer, add anhydrous sodium sulfate for dehydration, and concentrate the n-butanol layer under reduced pressure. The butanol part is the total glycosides extract of Xanthium fructus. At this time, the yield of the total glycosides extract of Xanthium is 1%-10%, that is, the total glycosides extract of Xanthium accounts for 1%-10% of the total weight of the medicinal materials of Xanthium.

③The separation and purification method adopted can also adopt the macroporous adsorption resin method. The macroporous adsorption resin method adopted can recover the total glycosides extract of Xanthium officinale medicinal material under reduced pressure until it has no alcohol smell, filter the filtrate, adjust the filtrate to a relative density of 1.0-1.1 with distilled water, and put it on a macroporous adsorption resin column, Water-soluble impurities are washed away with water, followed by eluting with eluting solvents such as ethanol aqueous solutions of different concentrations, and collecting 5-75% ethanol eluted parts to obtain an eluent rich in phenolic acids and diterpene glycosides. Recover to dryness under reduced pressure to obtain the total glycosides extract of Xanthium fructus. At this time, the yield of the total glycosides extract of Xanthium is 0.5%-10%, that is, the total glycosides extract of Xanthium accounts for 0.5%-10% of the total weight of the medicinal material of Xanthium. A preferred elution solvent is 40% ethanol in water.

④ The preparation method of the macroporous adsorption resin for the total glycosides extract of Xanthia fructus can use various types of macroporous adsorption resins, including D101, D520, D4006, H103, AB-8, Dianion HP 20 or XAD-7 and other nonpolar resins. One or more of polar and weak polar resins, or other fillers that can replace the above resins in performance, preferably one or more of AB-8, D101 or D520, etc., especially AB-8 .

All in all, in the preparation method of the total glycosides extract of Xanthium glucosides, fresh medicinal materials or commercially available medicinal materials of Xanthium are mainly used as raw materials; after the raw materials are pulverized, solvent extraction, solvent extraction or macroporous adsorption resin are sequentially used One or more methods, etc., to obtain the crude product of Xanthium glycosides extract; if combined with one or more of the commonly used drying methods such as reduced pressure concentration and drying, spray drying, freeze drying, etc., it can be prepared The extract or dry matter of the crude product of the total glycosides extract of Xanthium fructus.

If separation and purification methods such as column chromatography are further adopted, the phenolic acid compounds and diterpene glycosides in the extract of cocklebur glycosides can be obtained; at this time, the obtained phenolic acid compounds and diterpene glycosides Compounds account for 50% to 100% of the total glycosides extract of Xanthia glycosides, among which diterpene glycosides account for 0.5% to 99% (W/W) of the total glycosides extract of Xanthium glycosides, and phenolic acids account for 0.5% to 99% (W/W) of the glycoside extract content.

Taking several typical dosage forms as examples, the specific preparation methods of various dosage forms of the total glycosides extract of Xanthium fructus are described in detail below.

The powder injection preparation of the present invention generally adopts a conventional freeze-drying method, using water as a solvent, and the steps are: take the total glucosides extract of Xanthium fructus, add excipients, add water to dissolve, add activated carbon, filter and sterilize, fill, and semi- Stoppering, freeze-drying, stoppering and capping. The used excipient is selected from one or more of mannitol, hydrolyzed gelatin, glucose, lactose or dextran and the like. Each bottle contains 10-100 mg of total glycosides extract of Xanthium fructus.

The preparation powder injection of the present invention also can adopt spray-drying method, with water as solvent, and its steps are: take total glycosides extract of Xanthia fructus, add or not add excipient (excipient is the same as above), add water to dissolve, add activated carbon, Sterilize by filtration, spray-dry, aseptically dispense, and cork and cap. Each bottle contains 10-100 mg of total glycosides extract of Xanthium fructus.

When preparing small injections in the present invention, water for injection can be used as a solvent for preparation, and an appropriate amount of auxiliary materials can also be added. The auxiliary materials are selected from ethanol, propylene glycol, glycerin, polyethylene glycol, benzyl benzoate or dimethylacetamide species or several. Each tube contains 10-100 mg of total glycosides extract of Xanthium fructus.

The present invention prepares glucose infusion or sodium chloride infusion, uses water for injection as solvent, adds appropriate amount of glucose or sodium chloride to prepare, and can also add appropriate amount of auxiliary materials, and auxiliary materials are selected from ethanol, propylene glycol, glycerin, polyethylene glycol, benzoic acid One or more of benzyl ester or dimethylacetamide, etc. Each bottle contains 10-100 mg of total glycosides extract of Xanthium fructus.

The present invention prepares oral preparations such as tablets, capsules, granules, and oral liquids, and the auxiliary materials can be lactose, starch, dextrin, stearate, etc., and are prepared according to conventional techniques.

In the present invention, the specific implementation manners described above and the examples described below are for better illustrating the present invention, and are not used to limit the scope of the invention.

The present invention will be described in detail below by way of examples.

Embodiment 1, preparation method 1 of cocklebur glycosides extract

Xanthium 2.5Kg dried medicinal material, reflux extraction with 50 liters of 90% ethanol solution for 2 times, 2 hours each time, combine the two extracts, concentrate under reduced pressure to obtain 350 g of extract, disperse with 2 liters of water, and successively extract with 6 liters Petroleum ether was extracted and degreased three times, and impurities were extracted three times with 6 liters of chloroform and 6 liters of ethyl acetate respectively; the extraction residue was extracted three times with 6 liters of n-butanol, and the extracts were combined and freeze-dried to obtain the total glycosides of Xanthia glycosides. Material 21g.

Embodiment 2, preparation method 2 of cocklebur glycosides extract

Xanthium 2.5Kg dried medicinal material, ultrasonically extracted 2 times with 50 liters of 70% ethanol solution, 2 hours each time, combined the two extracts, concentrated under reduced pressure to obtain 208 g of extract, dispersed with 2 liters of water, and sequentially extracted in 6 liters Petroleum ether was extracted and degreased three times, and impurities were extracted three times with 6 liters of chloroform and 6 liters of ethyl acetate respectively; the extraction residue was extracted three times with 6 liters of n-butanol, and the extracts were combined and recovered to dryness under reduced pressure to obtain cocklebur Total glycosides extract 17g.

Embodiment 3, preparation method 3 of cocklebur glycosides extract

2.5Kg dried medicinal material of Fructus Xanthii, extracted twice by percolation with 50 liters of 75% ethanol solution, 24 hours each time, combined the two extracts, concentrated under reduced pressure to obtain 285g of extract, dispersed with 2 liters of water, and then washed with 6 1 liter of petroleum ether was extracted and degreased three times, respectively extracted three times with 6 liters of chloroform and 6 liters of ethyl acetate to remove impurities; the extraction residue was extracted three times with 6 liters of n-butanol, the combined extracts were recovered to dryness under reduced pressure to obtain Auricularia glycosides extract 19g.

Embodiment 4, preparation method 4 of cocklebur glycosides extract

Xanthium 2.5Kg dried medicinal material, reflux extraction with 50 liters of 75% ethanol solution for 2 times, each time for 2 hours, combine the two extracts, recover under reduced pressure until there is no alcohol smell, filter to obtain the filtrate, and adjust the filtrate to relative density with distilled water 1.05, put on D101 macroporous adsorption resin column, wash with water to remove water-soluble impurities, and then elute with different concentrations of ethanol, collect 50% ethanol eluted parts, that is, the elution rich in phenolic acid glycosides and diterpene glycosides liquid, and recovered to dryness under reduced pressure to obtain 15 g of total glucosides extract of Xanthium fructus.

Embodiment 5, preparation method 5 of total glycosides extract of Xanthium fructus

Xanthium 2.5Kg dried medicinal material, reflux extraction with 50 liters of 75% ethanol solution for 2 times, each time for 2 hours, combine the two extracts, recover under reduced pressure until there is no alcohol smell, filter to obtain the filtrate, and adjust the filtrate to relative density with distilled water 1.05, put on the AB-8 macroporous adsorption resin column, wash with water to remove water-soluble impurities, then elute with different concentrations of ethanol, collect 50% ethanol eluted parts, that is, the product rich in phenolic acid glycosides and diterpene glycosides The eluate was recovered to dryness under reduced pressure to obtain 20 g of total glycosides extract of Xanthium fructus.

Embodiment 6, preparation of total glycosides extract nasal drops of cocklebur

Take 45g of the total glycosides extract of Xanthium fructus obtained above, add 20g of pig gall paste, 0.5g of chlorpheniramine maleate, 8g of houttuyonin sodium, add distilled water to dilute 10 times, filter out the insoluble matter, and bottle it. Sterilize and seal to obtain the total glucoside extract nasal drops of cocklebur.

Example 7, preparation of cocklebur glycosides extract drop capsules

Take 40g of the total glycosides extract of Xanthium fructus obtained above, add pig gall paste 15, chlorpheniramine maleate 0.3g, sodium houttuyfonate 6g, add 40% to 60% starch, dextrin, carboxymethyl Starch, calcium hydrogen phosphate and other fillers, 0.5-10% disintegrant sodium carboxymethyl cellulose, hydroxypropyl cellulose are made into granules and dried, and glidant magnesium stearate and micro silica gel are added to make dried, and then packed into capsules to obtain the product, i.e. Xanthium glucosides extract capsules.

Example 8, Preparation of Xanthium glycosides extract capsules

(1) Prescription

Xanthium glycosides extract 1000.0g

Microcrystalline Cellulose 1000g

Sodium carboxymethyl starch 140g

Absolute ethanol Appropriate amount

Talc powder 80g

             

A total of 10000 capsules were made

(2) Preparation process According to the above prescription, the raw material Xanthium glycosides extract and other excipients in the prescription were respectively passed through a 100-mesh sieve, dried at 60°C, and the prescription amount of Xanthia glucosides extract and microcrystalline Cellulose and carboxymethyl starch sodium are mixed evenly in equal amounts, soft materials are made with appropriate amount of ethanol, granulated with a 30-mesh sieve, dried at 50-60°C for 2 hours, granulated with a 30-mesh sieve, and the prescribed amount of talc is added Mix powder and sodium carboxymethyl starch evenly.

Example 9, Preparation of Xanthium Total Glycosides Extract Powder Injection

Take 30g of total glycosides extract of Xanthia fructus, add 30g of dextran, add 500ml of water for injection, stir to dissolve; add water for injection to 2000ml, add 3.0g of activated carbon for needles, and stir for 30 minutes; decarbonize and filter; use 0.22 μm microporous membrane filtration; filling into sterile vials, 2ml per bottle, half-stoppered; freeze-dried, then stoppered and capped.

Example 10, Preparation of Xanthium Total Glycosides Extract Powder Injection

Take 60g of total glycosides extract of Xanthia fructus, add 500ml of water for injection, stir to dissolve; add water for injection to 1000ml, add 1g of activated carbon for needles, and stir thoroughly for 30 minutes; decarbonize and filter; filter with a 0.22μm microporous membrane; Freeze-dry to obtain sterile powder, and pack into 1000 bottles.

Example 11, Preparation of Xanthium Total Glycosides Extract Powder Injection

Take 40g of total glycosides extract of Xanthia fructus, add 50g of lactose, add 100ml of water for injection, stir to dissolve; add water for injection to 1000ml, add 1.5g of activated carbon for needles, and stir thoroughly for 30 minutes; decarbonize and filter; use 0.22μm micro Pore membrane filtration; spray-dried to get sterile powder, packed into 1000 bottles.

Example 12, Preparation of Xanthium Total Glycosides Extract Powder Injection

(1) Prescription

Xanthium glucosides extract semi-finished product for injection 150g

Mannitol 400g

Add water for injection to 10000ml

                 

A total of 10,000 bottles were made

(2) Preparation process

According to the above prescription, weigh the total glycosides extract of cocklebur glycosides in the prescribed amount, add it to an appropriate amount of water for injection, and stir to dissolve it; add the prescribed amount of mannitol, stir it to dissolve completely, add water for injection to the full amount; add 0.1% of the liquid amount Activated carbon for needles, fully stirred for 30 minutes; decarbonized filter; filtered with 0.22μm microporous membrane; filled, half-stoppered; freeze-dried, stoppered and capped. A total of 9735 bottles were produced, with a yield of 97.35%.

Example 13, Preparation of Small Injection of Total Glycosides Extract of Xanthium Fructus

Take 5g of total glycosides extract of Xanthia fructus, add 100ml of water for injection, stir to dissolve; add water for injection to 1000ml, filter with 0.22μm microporous membrane; subpackage and seal, 10ml per bottle, and sterilize.

Example 14, Preparation of Small Injection of Total Glycosides Extract of Xanthium Fructus

Take 10g of total glycosides extract of Xanthia fructus, add 30g of propylene glycol, add 200ml of water for injection, stir to dissolve; add water for injection to 1000ml, add 1.5g of activated carbon for needles, and stir thoroughly for 30 minutes; decarbonize and filter; use 0.22μm micro Pore membrane filtration; subpackaging and filling, 5ml per bottle, just sterilize.

Example 15, Preparation of Glucose Infusion of Total Glycosides Extract of Xanthium Fructus

Take 2g of total glycosides extract of Xanthia fructus, add 10g of polyethylene glycol, 500g of glucose, add 2000ml of water for injection, stir to dissolve; add water for injection to 5000ml; filter with 0.22μm microporous membrane; pack and seal , 100ml per bottle, just sterilized.

Example 16, Preparation of Glucose Infusion of Total Glycosides Extract of Xanthium Fructus

Take 2g of total glycosides extract of Xanthia fructus, add 250g of glucose, add 1000ml of water for injection, stir to dissolve; add water for injection to 5000ml; filter with 0.22μm microporous membrane; subpackage and seal, 250ml per bottle, and sterilize That's it.

Example 17, Preparation of Sodium Chloride Infusion of Total Glycosides Extract of Xanthia Fructus

Take 1g of total glycosides extract of Xanthia fructus, add 90g of sodium chloride, add 1000ml of water for injection, stir to dissolve; add water for injection to 10000ml; filter with 0.22μm microporous membrane; subpackage and seal, 250ml per bottle, Just sterilize.

Example 18, preparation of Xanthium glycosides extract tablet

Take 80g of total glycosides extract of Xanthia fructus, 80g of microcrystalline cellulose, 15g of lactose, and 60g of pregelatinized starch, sieve, mix evenly, use an appropriate amount of 10% PVP ethanol solution to make a soft material, granulate, dry, and add stearin Magnesium acid 3g, whole grain, compressed into tablets, made into 1000 tablets.

Example 19, preparation of Xanthium glycosides extract tablet

(1) Prescription

Xanthium glycosides extract 1000.0g

Microcrystalline Cellulose 1170.0g

Pregelatinized starch 690.0g

Lactose 125.0g

5% PVP absolute ethanol Appropriate amount

Magnesium stearate 15.0g

             

A total of 10,000 pieces were made

(2) Preparation process

According to the above prescription, weigh the main drug and auxiliary materials of the prescription amount, mix them evenly according to the method of equal volume addition, and then carry out the processes of making soft materials, granules, drying, and granulation according to the prescription process items, and after calculating the tablet weight, use single punching A tablet machine and a 10.5mm shallow concave die were used to press the tablet, and the hardness of the bare chip was controlled to be 5-7kg. A total of 9698 tablets were produced, with a yield of 96.98%. Rolling spray coating is adopted, and the coating process is as follows:

Preparation of coating solution: gastric soluble film coating material: 85G61235, provided by Shanghai Colorcon Coating Technology Co., Ltd.

Coating process: Put the bare chip to be coated (hardness 5kg~7kg) into the coating pot, start the stirring device and the blast heating device, and when the temperature of the bare chip rises to 40°C, start to open the spray gun and aim at the tablet bed. Spray the coating solution into the upper 1/3 of the tablet, control the tablet bed temperature at 38°C to 42°C, the gas pound pressure at 6kg, the flow rate of the coating solution at 50mL/min, and the weight of the coating film accounts for 3% of the weight of the coated tablet.

Example 20, the analgesic effect of the total glycosides extract of Xanthium fructus

An appropriate amount of the total glucosides extract of Xanthia fructus (abbreviated as CE) obtained above was taken, and the analgesic effect of total glucosides of Xanthia fructus was observed by using the mouse writhing reaction model induced by acetic acid.

(1) Experimental method

Take 60 mice of 20±2g, half male and half male, and randomly group them. Each group was administered continuously for 3 days by the dosage in Table 6 respectively, and 0.7% glacial acetic acid solution (0.1ml/10g mouse) was injected intraperitoneally 30 minutes before the last administration, and the number of times of mouse writhing in 10 minutes was observed and recorded after 5 minutes, and the results are shown in the table 3.

(2) Experimental results

The results in Table 3 show that each CE group can significantly reduce the number of writhing reactions in mice induced by acetic acid, and has an analgesic effect.

Table 3. Effect of CE on the writhing response of mice induced by acetic acid (X±SD, n=10)

group Dosage Route of administration   Number of twists Normal saline group 2ml/kg Ig 21.2±11.0 Antipyretic and Pain Relief Powder 0.1g/kg Ig 8.5±6.6 ^** CE group 10g/kg Ig 8.0±5.3 ^** 5.0g/kg Ig 8.8±7.0 ^** 2.5g/kg Ig 11.9±7.8 ^*

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

Example 21, Therapeutic effect of cocklebur glycosides extract on experimental sinusitis

Take an appropriate amount of the total glucosides extract (abbreviation: CE) obtained above, and use a rabbit experimental sinusitis model to observe the effect of total glycosides from Xanthia glucosides on the histomorphology of the nasal mucosa of the rabbits with a light microscope.

(1) Experimental method

Select 56 healthy New Zealand white rabbits weighing about 2.5kg, half male and half male, and divide them into random groups. On the first day, under general anesthesia, the ostium of the left maxillary sinus was opened, gelatin sponge was used to block the ostium of the unilateral sinus, and surgical suturing was performed. The whole process is carried out aseptically. On the next day, 0.5ml of 108 CFU/ml Streptococcus pneumoniae suspension was injected, and the administration group was continuously combined with the medicine for 10 days according to the dose in Table 1. After the last administration, the cells were separated and cultured, materials were drawn, and slices were made. The following indicators were observed under a light microscope: ①The thickness of the nasal mucosa. To facilitate positioning and comparison, the thickness of each specimen from the apex of the mucosal bulge to the side edge of the nasal septal cartilage (including the epithelial layer and lamina propria) was measured with an eyepiece micrometer as the thickness of the mucosa; ②The number of glands in the lamina propria is based on the average number of glands under the bilateral mucosal protrusion; ③The number of goblet cells in the mucosal epithelium is based on the average of the total number of goblet cells randomly counted in 5 high-power fields;④ The number of lymphoid nodules in the lamina propria is the total number of lymphoid nodules in the section.

(2) Experimental results

The morphological changes of nasal mucosa in each group were observed and analyzed, and the results are shown in Table 4.

The results in Table 3 and Table 4 show that the middle and high dose groups of CE significantly reduced the infiltration of nasal mucositis cells, eliminated nasal mucosal edema, inhibited the hyperplasia of submucosal connective tissue, and reduced the incidence of nasal mucosal epithelial goblet cells and glandular hyperplasia in the lamina propria. The effect and histomorphological indicators were generally stronger than those of the levofloxacin group, and the high-dose group was stronger than that of the Biyuanshu group.

Table 4. Effect of CE on histomorphological indexes of sinus mucosa in rabbits (X±SD, n=8)

group dose Route of administration Mucosa thickness (m) The number of submucosal protruding glandular ducts (pieces) Mucosal epithelial goblet cells (units) The number of lymph nodes (pieces) normal group - - 0.185±0.036 7.71±1.50 5.38±1.69 2.00±0.82 model group - - 0.402±0.120 17.86±1.95 14.00±2.83 7.29±3.86 Levofloxacin group 120mg/kg ig 0.388±0.083 12.58±2.63 ^* 12.34±3.22 5.77±1.98 ^* Biyuanshu group 2ml/kg ig 0.257±0.060 ^** 10.44±1.29 ^* 10.56±1.88 ^* 4.26±0.95 ^* CE group 10g/kg ig 0.230±0.035 ^** 8.71±1.75 ^** 6.88±1.64 ^** 3.14±0.57 ^** 5.0g/kg ig 0.260±0.087 ^** 8.85±2.06 ^** 7.02±1.59 ^** 3.45±1.08 ^* 2.5g/kg ig 0.343±0.108 10.08±2.46 ^* 11.23±1.20 ^* 4.54±0.80 ^*

Compared with the model group, ^* P<0.05, ^** P<0.01

Example 22, Inhibition of Influenza Virus by Extracting Total Glycosides of Xanthium Fructus

(1) Experimental method

① MDCK (Madin darby canin kidney) cell subculture and influenza virus (Influenza A1 virus) culture: according to the literature (Guo Yuanji, Cheng Xiaowen; 1997) method.

②Cytotoxicity test: add the sample to the cell plate that has grown into a single layer of cells at 0.1mL/well, and add cell maintenance solution to 1mL/well, culture in a 37°C, 5% CO2 incubator for 72h, and observe the cytopathic changes. At the same time, MDCK cells were set as control. The experiment was repeated 2 times. The results showed that the sample did not produce non-specific cytopathic effect (CPE) on MDCK cells.

③ Sample anti-influenza virus test: cells were cultured in 96-well MDCK cell culture plate, and cell control group, virus control group, positive control group and test group were respectively set up. Influenza A1 virus was added to the virus control group and the test group, adsorbed at 37°C for 2 hours, and the virus was aspirated. Samples with different concentrations were added to each test group, cultured at 37°C and 5% CO2 for 3 days, the test results were observed, and the 50% inhibitory concentration (IC ₅₀ ) of different drugs on the virus was calculated.

(2) Experimental results (see: Table 5)

The experimental results showed that the total glycosides extract of Xanthium Fructus had obvious inhibitory effect on influenza virus.

Table 5. Inhibitory effect of total glycosides extract from Xanthium Fructus on influenza virus

Group _IC50 (mmol/L) CE 4.2 Ribavirin 3.9

Claims (34)

1. The application of the composition of the total glycosides extract of Xanthia fructus in the preparation of anti-inflammatory reaction products, and the anti-inflammatory reaction products refer to products for anti-inflammation and related diseases, products for anti-virus and related diseases , products for pain relief and related conditions, or one or more of products for the diagnosis, detection, treatment or research of sinusitis and related conditions. 2. The application of the composition of the total glycosides extract of Xanthia fructus according to claim 1, characterized in that, the anti-inflammatory reaction product comprises one or more of drugs, reagents, food or beverages . 3. The application of the composition of the total glycosides extract of Xanthia fructus according to claim 2, characterized in that, the anti-inflammatory reaction product is a medicine for diagnosis, detection, treatment or research of sinusitis and related diseases. 4. the application of the composition of total glycosides extract of Xanthium fructus according to claim 1, is characterized in that, said total glycosides extract comprises diterpene glycosides, phenolic acid compounds and other components. total extract. 5. the application of the composition of total glycosides extract of Xanthium fructus according to claim 4, is characterized in that, described diterpene glycosides compound, phenolic acid compound and other compositions in this total glycosides extract The content ranges are respectively 0.5%-99% (weight percentage), 0.5%-99% and 0.5% or less. 6. the application of the composition of total glycosides extract of Xanthium fructus according to claim 5, is characterized in that, the content scope of described diterpene glycosides compound and phenolic acid compound in this total glycosides extract respectively It is 5% to 95% and 5% to 95%. 7. the application of the composition of total glycosides extract of Xanthium fructus according to claim 4, is characterized in that, the general chemical structure formula of diterpene glycosides is as follows:

Wherein, R ₁ is one of hydrogen, methyl or carboxyl; R ₂ and R ₃ are one of hydrogen, sulfonic acid, acetyl, propionyl or dimethylpropionyl. 8. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 7, wherein said diterpene glycosides comprise one or more of the following compounds: When R ₁ =COOH, R ₂ =R ₃ =H, it means diterpene glycoside compound A; When R ₁ =COOH, R ₂ =H, R ₃ =COCH ₂ CH(CH ₃ ) ₂ , it means diterpene glycoside compound B; When R ₁ ＝COOH, R ₂ ＝SO ₃ ^- , R ₃ ＝COCH ₂ CH(CH ₃ ) ₂ , it means diterpene glycoside compound C; When R ₁ ＝H, R ₂ ＝H, R ₃ ＝COCH ₂ CH(CH ₃ ) ₂ , it means diterpene glycoside compound D; When R ₁ =H, R ₂ =SO ₃ ^- , R ₃ =COCH ₂ CH(CH ₃ ) ₂ , it means diterpene glycoside compound E. 9. The application of the composition of the total glucosides of Xanthium glucosides extract according to claim 4, is characterized in that, the general chemical structure formula of described phenolic acid compounds is as follows:

Wherein, R ₁ is one of hydrogen or an alkali metal element; R ₂ , R ₃ and R ₄ are one of hydrogen or caffeoyl. 10. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 9, wherein the phenolic acid compound comprises one or more of the following compounds: When R ₁ = H, R ₂ = R ₃ = R ₄ = caffeoyl, it means phenolic acid compound A; When R ₁ =R ₃ =H, R ₂ =R ₄ =caffeoyl, it means phenolic acid compound B; When R ₁ =R ₂ =H, R ₃ =R ₄ =caffeoyl, it means phenolic acid compound C; When R ₁ =K, R ₂ =R ₄ =H, R ₃ =caffeoyl, it represents the phenolic acid compound D. 11. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 4, characterized in that, the preparation method of the total glycosides extract of Xanthium fructus comprises the following steps: (1) Extraction: get some raw materials of Fructus Xanthium, extract, and obtain the extract, i.e. the total extract of Fructus Xanthium; (2) Separation and purification: Separation and purification of the extract to obtain the total glycosides extract of Xanthium fructus. 12. The application of the composition of the total glycosides extract of Xanthia fructus according to claim 11, characterized in that, the extraction method is all available methods known in the art including solvent extraction. 13. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 12, characterized in that, the extraction method is a solvent extraction method. 14. The application of the composition of the total glucosides extract of Xanthium fructus according to claim 13, is characterized in that, described solvent extraction method comprises common ultrasonic extraction method, dipping extraction method, percolation extraction method, decocting One or more of boiling extraction, heating reflux extraction or continuous reflux extraction; the number of extractions is one or more. 15. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 14, characterized in that, the solvent extraction method comprises one of ultrasonic extraction, percolation extraction or heating reflux extraction. species; the number of extractions is multiple. 16. The application of the composition of the total glucosides of Xanthia glucosides extract according to claim 15, is characterized in that, the used extraction solvent of described solvent extraction method is to comprise common water reagent, hydrophilic organic solvent Or one or more of the three types of extraction solvents in lipophilic organic solvents. 17. The application of the composition of the total glycosides extract of Xanthia fructus according to claim 16, characterized in that, the water reagent is one of water, acid water or alkaline water; the hydrophilic The organic solvent comprises one or more of ethanol, ethanol aqueous solution or methanol; the lipophilic organic solvent comprises petroleum ether, chloroform, ether, ethyl acetate, dichloromethane or dichloroethane one or more. 18. The application of the composition of the total glycosides extract of Fructus Xanthium according to claim 11, characterized in that, the extraction method is: take some Fructus Xanthium medicinal materials, add extraction solvent and mix, extract, and extract Filter and concentrate to obtain the total extract of Xanthium medicinal material. 19. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 18, characterized in that, the extraction solvent includes one or more of water, alcohols or other organic solvents. 20. The application of the composition of the total glycosides extract of Xanthium Fructus according to claim 19, characterized in that, the alcohol extraction solvent is one of methanol, ethanol, aqueous ethanol, propanol or butanol One or more, the other organic solvents include one or more of ethyl acetate or acetone. 21. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 20, characterized in that, the alcohol extraction solvent is 60%-90% ethanol aqueous solution. 22. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 11, characterized in that, the separation and purification method comprises solvent separation, solvent extraction, macroporous adsorption resin method, precipitation method , salting-out method, column chromatography or one or more of crystallization and recrystallization and fractional crystallization. 23. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 22, characterized in that, the solvent extraction method comprises extraction method, countercurrent continuous extraction method, countercurrent distribution method or droplet countercurrent distribution one or more of the laws. 24. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 23, is characterized in that, the operation step of described solvent extraction method is: (1) Disperse the total extract of Xanthium officinalis with water, degrease with petroleum ether, and then extract with an appropriate organic solvent to remove fat-soluble components to obtain an extraction residue; the number of extractions can be one or more times. (2) Continue to extract the extraction residue with n-butanol, separate the n-butanol layer, add anhydrous sodium sulfate for dehydration, and concentrate the n-butanol part under reduced pressure to obtain the total glycosides extract of Xanthia fructus. 25. The application of the composition of the total glycosides extract of Fructus Xanthium according to claim 23, characterized in that, the operation of the solvent extraction method comprises the following steps: directly using an appropriate organic solvent to extract the total glycosides of Fructus Xanthium. The extract is subjected to extraction to remove fat-soluble components to obtain an extraction residue; the number of extractions may be one or more. 26. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 25, characterized in that, the appropriate organic solvent comprises one of chloroform, dichloromethane, ether or ethyl acetate or more. 27. The application of the composition of the total glucosides extract of Xanthium fructus according to claim 22, is characterized in that, the operation of described macroporous adsorption resin method comprises the following steps: (1) Recover the total glycoside extract of Xanthium officinalis medicinal material under reduced pressure until it has no alcohol smell, and filter to obtain the filtrate; (2) Adjust the filtrate to a relative density of 1.0 to 1.1 with distilled water, put it on a macroporous adsorption resin column, and wash away water-soluble impurities with water; (3) elution is performed with an elution solvent, and 5-75% alcohol elution sites are collected to obtain an eluate rich in phenolic acids and diterpene glycosides; (4) Recover to dryness under reduced pressure to obtain the total glycosides extract of Xanthium fructus. 28. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 27, characterized in that, the eluting solvents are aqueous ethanol solutions of different concentrations. 29. The application of the composition of the extract of total glycosides of Xanthia fructus according to claim 28, characterized in that, the eluting solvent is an aqueous ethanol solution with a concentration of 40%. 30. The application of the composition of the extract of total glycosides of Xanthium fructus according to claim 27, characterized in that, the macroporous adsorption resin includes various types of macroporous adsorption resins. 31. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 30, characterized in that, the macroporous adsorption resin includes non-polar resin, weak polar resin or other properties capable of One of the fillers to replace the above resins. 32. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 31, characterized in that, the non-polar resin and weak polar resin include D101, D520, D4006, H103, AB- 8. One of Dianion HP 20 or XAD-7. 33. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 11, further comprising a drying step. 34. The application of the composition of the total glycosides extract of Xanthium fructus according to claim 33, characterized in that, the drying method comprises normal pressure drying method, reduced pressure drying method, vacuum drying method, spray drying method , freeze-drying, far-infrared heating and drying or one or more of microwave drying.