WO2016082703A1 - 一种包含绿原酸晶型的制剂及其用途 - Google Patents

一种包含绿原酸晶型的制剂及其用途 Download PDF

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WO2016082703A1
WO2016082703A1 PCT/CN2015/094869 CN2015094869W WO2016082703A1 WO 2016082703 A1 WO2016082703 A1 WO 2016082703A1 CN 2015094869 W CN2015094869 W CN 2015094869W WO 2016082703 A1 WO2016082703 A1 WO 2016082703A1
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chlorogenic acid
preparation
weight
parts
tumor
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French (fr)
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张洁
朱丽娜
黄望
张亮
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四川九章生物科技有限公司
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Priority to JP2017527880A priority Critical patent/JP2017537092A/ja
Priority to US15/528,996 priority patent/US10314806B2/en
Priority to EP15864107.6A priority patent/EP3225234A4/en
Publication of WO2016082703A1 publication Critical patent/WO2016082703A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/216—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acids having aromatic rings, e.g. benactizyne, clofibrate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system

Definitions

  • the invention relates to a preparation comprising a crystalline form of chlorogenic acid and a use thereof, and belongs to the field of pharmaceutical preparations.
  • Chlorogenic acid a depsipeptide produced from caffeic acid and quinic acid, is a phenylpropanoid compound produced by the plant in the aerobic respiration process by the shikimic acid pathway.
  • Chlorogenic acid has a wide range of biological activities, and modern scientific research on the biological activity of chlorogenic acid has penetrated into many fields such as food, health care, medicine and daily chemical industry. Chlorogenic acid is an important biologically active substance with antibacterial, antiviral, white blood cells, liver and gallbladder, anti-tumor, blood pressure lowering, blood lipid lowering, free radical scavenging and excitatory central nervous system.
  • chlorogenic acid has a variety of crystal forms, but the comparative study on the efficacy of chlorogenic acid crystal form preparation has not been reported. Because different crystal forms of the same pharmaceutical preparation often cause significant differences in physical and chemical properties such as appearance, solubility, melting point, density, and drug dissolution and bioavailability, thereby affecting the effects of drug stability and bioavailability.
  • the anti-ulcer drug famotidine has four crystal forms, and its melting point, infrared spectrum and physical and chemical properties are obviously different, and the active B type which inhibits gastric acid secretion is larger than type A. Some drugs have a crystalline state that is not as good as an amorphous one, such as an amorphous hygromycin suspension.
  • the chlorogenic acid crystal form content in the preparation is preferably from 10 to 500 mg/g, depending on the dosage form.
  • the dosage form of the preparation may be a conventional dosage form in the art including, but not limited to, an oral preparation, an injection, a respiratory administration preparation, a dermal administration preparation, a mucosal administration preparation, and the like.
  • the Applicant has found through research that controlling the crystal form of chlorogenic acid in chlorogenic acid preparations can significantly improve the use effect. Further, the Applicant has found that in the injection preparation, when the chlorogenic acid crystal form content is 10-60 mg/kg, the preparation has a tumor for human glioma, liver cancer, lung cancer, breast cancer and the like. Significant inhibition has a more excellent effect, preferably 20-40 mg/kg. According to the dose conversion relationship between animals and humans, the clinical use dose protection of chlorogenic acid freeze-dried powder injection: 1.0-6.0 mg/kg per day, preferably 2-4 mg/kg.
  • the Applicant has found that in the oral preparation, when the chlorogenic acid crystal form content is 20-200 mg/kg, the preparation has obvious protective effect against the liver, and has a more excellent effect, preferably 50-140 mg/kg. .
  • the clinically used dosage range of the chlorogenic acid oral preparation is from 2 mg to 20 mg/kg per day, preferably from 5 to 14 mg/kg, depending on the dose-converting relationship between the animal and the human.
  • the oral preparation and the injection are prepared from chlorogenic acid or chlorogenic acid and a pharmaceutically acceptable pharmaceutical excipient.
  • the crystal form of the chlorogenic acid can be prepared by the method described in Chinese Patent Application No. CN201410193699.8, which can also be carried out by the method described in Chinese Patent Application No. CN201310366945.0.
  • the lyophilized powder injection comprises the following components:
  • the lyophilized powder injection has a chlorogenic acid crystal of 30 to 40 parts by weight; the antioxidant is preferably 2 to 5 parts by weight; and the scaffolding agent is preferably 80 to 90 parts by weight.
  • the antioxidant is sodium bisulfite and the scaffold is mannitol.
  • the lyophilized powder injection comprises the following components per 1000 formulation units:
  • oral preparation of the present invention comprises the following components:
  • the binder is 20-220 parts by weight.
  • the chlorogenic acid crystal is preferably 80 to 100 parts by weight; the filler is preferably 800 to 900 parts by weight; and the binder is preferably 100 to 220 parts by weight.
  • the filler includes, but is not limited to, one or more of starch, icing sugar, dextrin, lactose, compressible starch, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate or mannitol.
  • the binder includes, but is not limited to, one or more of sodium carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, ethylcellulose, polyvinylpyrrolidone.
  • the oral formulation comprises the following components per 1000 formulation units:
  • Another object of the present invention is to provide an application of the above preparation in the preparation of a medicament for treating diseases such as anti-tumor, psoriasis and the like, anti-oxidation, liver and gallbladder, cardiovascular disease, anti-virus, etc., Including, but not limited to, small cell lung cancer, liver cancer, breast cancer, brain tumors, psoriasis.
  • the invention has the beneficial effects that the chlorogenic acid crystal form preparation of the invention is more advantageous for the therapeutic effect of the bioavailability of the drug, the dominant crystal form for the chlorogenic acid, and the anti-tumor, psoriasis and the like immunity.
  • the use of drugs for systemic diseases, anti-oxidation, liver and gallbladder, cardiovascular diseases, anti-virus and the like exerts a more excellent clinical effect, and the present invention provides an effective dose for the clinical application of the chlorogenic acid crystal form preparation. It is a safe and effective basis for the clinical application of the chlorogenic acid crystal form preparation.
  • Figure 1 is a graph showing the effect of chlorogenic acid and commercially available chlorogenic acid on the tumor inhibition rate of Lewis lung cancer C57BL/6 mice.
  • Figure 3 is a graph showing the effect of chlorogenic acid and commercially available chlorogenic acid on the tumor inhibition rate of transplanted tumors of EMT-6 breast cancer BABLc mice.
  • Figure 4 is a graph showing the effect of chlorogenic acid and commercially available chlorogenic acid on the tumor inhibition rate of human glioma mice.
  • the prescription is determined according to 1000 preparation units (30 mg/piece):
  • the raw materials are weighed, and the injection water is sequentially added with an antioxidant (such as sodium hydrogen sulfite), chlorogenic acid crystals, a scaffold (such as mannitol), stirred to fully dissolve, and the pH is controlled at 2-4;
  • an antioxidant such as sodium hydrogen sulfite
  • chlorogenic acid crystals such as mannitol
  • a scaffold such as mannitol
  • the temperature is 45-50 ° C, 0.03% of activated carbon is added, stirred for 30 minutes, filtered to remove activated carbon, and then finely filtered with a 0.22 micron hydrophilic microfiltration membrane until the filtrate is clarified, aseptically filled, and lyophilized.
  • the prescription is determined according to 1000 preparation units (100 mg/tablet):
  • the chlorogenic acid crystal form is added to an appropriate amount of the filler and after the binder, the mixture is sieved, granulated, sized, and an appropriate amount of the lubricant is added and compressed.
  • Cell line Lewis cells are mouse lung cancer cell lines, adherently grown in RPMI-1640 medium containing 10% calf serum, 100 U ⁇ ml -1 penicillin, 100 ⁇ g ⁇ ml -1 streptomycin at 37 ° C, 5 Incubate in %C0 2 incubator and change it every 2 to 3 days.
  • the cell culture cells were routinely resuscitated and passaged.
  • the Lewis cells in the logarithmic growth phase were digested with 0.25% trypsin, the digestion was terminated, RPMI-1640 medium was added, the cells were blown to make a suspension, and centrifuged at 1000 rpm for 5 minutes, washing 2 The number of viable cells was counted by trypan blue staining.
  • the cell suspension was inoculated subcutaneously into the left forelimb of the mouse, 0.2 ml ⁇ only -1 (about 1 ⁇ 10 7 cells), and the mice were used as experimental mice when the tumor grew 1 cm ⁇ 1 cm ⁇ 1 cm.
  • mice were aseptically stripped and washed with Hank's solution for 3 times to remove blood stains, fat and necrotic tissue.
  • the tumor was cut into pieces of 1 mm ⁇ 1 mm ⁇ 1 mm, and Hank's solution was washed twice, and physiological saline (1 g) was added in proportion. : 3 ml), and then ground in a glass homogenizer, and filtered through an 80-100 mesh sieve to prepare a single cell suspension, and the number of viable cells was counted by trypan blue staining.
  • the prepared cell suspension was inoculated into the left anterior axilla of C57BL/6 mice by 0.2 ml ⁇ only -1 (about 1 ⁇ 10 7 cells), and randomly divided into groups according to body weight, 10 in each group, respectively, green.
  • Oral acid dose group 100mg ⁇ kg -1 , 90mg ⁇ kg -1 , 80mg ⁇ kg -1 , 70mg ⁇ kg -1 , 60mg ⁇ kg -1 , 50mg ⁇ kg -1 , 40mg ⁇ kg -1 , 30mg ⁇ Kg -1 , 20 mg ⁇ kg -1 , 10 mg ⁇ kg -1 , 5 mg ⁇ kg -1 , 1 mg ⁇ kg -1 and negative group (NS saline).
  • each group was intraperitoneally injected (ip) with a volume of 0.2 ml ⁇ 10 g -1 once daily for 12 consecutive administrations.
  • the average tumor size of the negative group was greater than 1.0 g (the tumor volume was about 0.5 cm 3 )
  • the experiment was stopped, the mice were sacrificed by cervical dislocation and weighed, the tumor was removed, and the tumor inhibition rate was calculated.
  • Tumor inhibition rate % [1 - (average tumor weight of the administration group / mean tumor weight of the negative group)] ⁇ 100%.
  • the chlorogenic acid of the present invention significantly inhibited the transplanted tumor of Lewis lung cancer C57BL/6 mice, and compared with the negative control group, there was a statistical difference. And the dose-fixing effect was obvious at 10 mg/kg-60 mg/kg.
  • Cell line H22 cell line is mouse hepatoma cells, which are suspended in RPMI-1640 medium and contain 10% calf serum, 100 U ⁇ ml -1 penicillin and 100 ⁇ g ⁇ ml -1 streptomycin at 37 ° C, 5%. C0 2 incubator culture, every 2 to 3 days change the liquid and pass it once.
  • the tumor was cut into 1 mm ⁇ 1 mm ⁇ 1 mm pieces, Hank's solution was washed twice, and physiological saline was added in proportion (1 g: 3 ml). Then, it was ground in a glass homogenizer, filtered through an 80-100 mesh screen to prepare a single cell suspension, and the number of viable cells was counted by trypan blue staining.
  • the prepared cell suspension was inoculated into the left anterior axilla of KM mice by 0.2 ml ⁇ only -1 (about 1 ⁇ 10 6 cells), and randomly divided into groups according to body weight, each group consisting of 10 chlorogenic acids.
  • dose group 100mg ⁇ kg -1, 90mg ⁇ kg -1, 80mg ⁇ kg - 1, 70mg ⁇ kg -1, 60mg ⁇ kg -1, 50mg ⁇ kg -1, 40mg ⁇ kg -1, 30mg ⁇ kg - 1 , 20 mg ⁇ kg -1 , 10 mg ⁇ kg -1 , 5 mg ⁇ kg -1 , 1 mg ⁇ kg -1 and the negative group (NS saline).
  • the intravenous (iv) administration was started on the second day after the inoculation, and the volume was 0.2 ml ⁇ 10 g -1 once a day.
  • the average tumor size of the negative group was greater than 1.0 g (the tumor volume was about 0.5 cm 3 )
  • the experiment was stopped, the mice were sacrificed by cervical dislocation and weighed, the tumor was removed, and the tumor inhibition rate was calculated.
  • the chlorogenic acid of the present invention has a significant inhibitory effect on the transplanted tumor of H22 liver cancer KM mice, and is negative. There was a statistical difference between the control groups. And the dose-fixing effect was obvious at 10 mg/kg-60 mg/kg.
  • Example 5 In vivo study on inhibition of transplanted tumor in BABLc mice
  • Cell line EMT-6 cell line is mouse breast cancer cells. Adherent growth in RPMI-1640 complete medium containing 10% calf serum, 1mmol/L glutamine, 100U ⁇ ml -1 penicillin and 100 ⁇ g ⁇ ml -1 in 37 ° C, 5% CO 2 incubator Culture, change the liquid every 2 to 3 days.
  • EMT-6 cells in logarithmic growth phase were digested with 0.25% trypsin, the digestion was terminated, RPMI-1640 medium was added, and the cells were blown to make a suspension, and centrifuged at 1000 rpm. Minutes, washed twice, and the number of viable cells was counted by trypan blue staining.
  • the cell suspension of the left forelimb of the mouse was inoculated subcutaneously with 0.2 ml ⁇ only-1 (about 1 ⁇ 107 cells), and the mice were used as experimental mice when the tumor grew 1 cm ⁇ 1 cm ⁇ 1 cm.
  • mice were aseptically stripped and washed with Hank's solution for 3 times to remove blood stains, fat and necrotic tissue.
  • the tumor was cut into 1 mm ⁇ 1 mm ⁇ 1 mm pieces, Hank's solution was washed twice, and physiological saline was added in proportion (1 g: 3 ml). Then, it was ground in a glass homogenizer, filtered through an 80-100 mesh screen to prepare a single cell suspension, and the number of viable cells was counted by trypan blue staining.
  • the prepared cell suspension was inoculated into the left anterior axilla of 70 BABLc mice by 0.2 ml ⁇ only -1 (containing about 1 ⁇ 106 cells), and randomly divided into groups according to body weight, 10 mice in each group, respectively.
  • Acid administration dose group 100mg ⁇ kg -1 , 90mg ⁇ kg -1 , 80mg ⁇ kg -1 , 70mg ⁇ kg -1 , 60mg ⁇ kg -1 , 50mg ⁇ kg -1 , 40mg ⁇ kg -1 , 30mg ⁇ kg -1 , 20 mg ⁇ kg -1 , 10 mg ⁇ kg -1 , 5 mg ⁇ kg -1 , 1 mg ⁇ kg -1 and a negative group (NS saline).
  • each group was administered intramuscularly (ip) with a volume of 0.2 ml ⁇ 10 g -1 once daily for 9 days.
  • the average tumor size of the negative group was greater than 1.0 g (the tumor volume was about 0.5 cm 3 )
  • the experiment was stopped, the mice were sacrificed by cervical dislocation and weighed, the tumor was removed, and the tumor inhibition rate was calculated.
  • Tumor inhibition rate % [1 - (average tumor weight of the administration group / mean tumor weight of the negative group)] ⁇ 100%.
  • Chlorogenic acid of the invention 100 10 0.995 ⁇ 0.0256 25.80
  • Chlorogenic acid of the invention 90 10 0.859 ⁇ 0.0754 35.94
  • Chlorogenic acid of the invention 80 10 0.789 ⁇ 0.087 41.16
  • Chlorogenic acid of the invention 70 10 0.698 ⁇ 0.112*** 47.95 Chlorogenic acid of the invention 60 10 0.681 ⁇ 0.156*** 49.22
  • Chlorogenic acid of the invention 50 10 0.529 ⁇ 0.168*** 60.55
  • Chlorogenic acid of the invention 40 10 0.451 ⁇ 0.097*** 66.37 Chlorogenic acid of the invention 30 10 0.429 ⁇ 0.168*** 68.01 Chlorogenic acid of the invention 20 10 0.403 ⁇ 0.101*** 69.95 Chlorogenic acid of the invention 10 10 0.690 ⁇ 0.188*** 48.55 Chlorogenic acid of the invention 5 10 0.789 ⁇ 0.087 41.16 Chlorogenic acid of the invention 1 10 0.987 ⁇ 0.154 26.40 Commercially available chlorogenic acid 100 10 1.312 ⁇ 0.087 2.16 Commercially available chlorogenic acid 90 10 1.298 ⁇ 0.423 3.21 Commercially available chlorogenic acid 80 10 1.279 ⁇ 0.143 4.62 Commercially available chlorogenic acid 70 10 1.254 ⁇ 0.514 6.49 Commercially available chlorogenic acid 60 10 1.009 ⁇ 0.123 24.76 Commercially available chlorogenic acid 50 10 1.115 ⁇ 0.201 16.85 Commercially available chlorogenic acid 40 10 1.125 ⁇ 0.125 16.11 Commercially available chlorogenic acid 30 10 1.185 ⁇ 0.247 11.63 Commercially available chlorogenic acid 20 10 1.201 ⁇ 0.546 10.44
  • the chlorogenic acid of the present invention has a significant inhibitory effect on the transplanted tumor of EMT-6 breast cancer BABLc mice, and is statistically different from the negative control group. And the dose-fixing effect was obvious at 10 mg/kg-60 mg/kg.
  • Example 6 Inhibition of chlorogenic acid on transplanted glioma
  • Modeling Mice were randomly divided into groups of 10 each.
  • the glioblastoma cell line was dosed into a suitable dilution and inoculated into the left temporal cortex of the mouse.
  • each group of mice was intraperitoneally administered.
  • Chlorogenic acid 100mg ⁇ kg -1 , 90mg ⁇ kg -1 , 80mg ⁇ kg -1 , 70mg ⁇ kg -1 , 60mg ⁇ kg -1 , 50mg ⁇ kg -1 , 40mg ⁇ kg -1 , 30mg ⁇ kg -1 , 20 mg ⁇ kg -1 , 10 mg ⁇ kg -1 , 5 mg ⁇ kg -1 , 1 mg ⁇ kg -1 , and the negative control group was given the same volume of physiological saline for 15 days.
  • Inhibition rate (average tumor weight of the control group - mean tumor weight of the treatment group) / tumor weight control group mean tumor weight ⁇ 100%.
  • the chlorogenic acid of the present invention has a significant inhibitory effect on transplanted tumors of human glioma mice, and is statistically different from the negative control group. And the dose-fixing effect was obvious at 10 mg/kg-60 mg/kg.
  • Example 7 Protective effect of chlorogenic acid on chemical liver injury (alcoholic fatty liver rat model)
  • the chlorogenic acid of the present invention is provided by Sichuan Jiuzhang Biotechnology Co., Ltd. Commercially available chlorogenic acid.
  • Animal Female SD rats weighing 156-193 g, SPF grade.
  • Test group and dose design Female SD rats were randomly divided into groups of 10 animals, and the experiment was set at 20 mg/kg, 35 mg/kg, 50 mg/kg, 70 mg/kg, 85 mg/kg, 100 mg/kg, 120 mg/kg, The 140 mg/kg, 160 mg/kg, 180 mg/kg, and 200 mg/kg dose groups were additionally provided with a negative control group and a 50% ethanol model control group. A model of liver injury was induced with ethanol (analytical grade).
  • Test method Alcoholic liver injury model was used.
  • the chlorogenic acid group was intragastrically administered with different doses of the test drug.
  • the negative control group and the model control group were given normal saline.
  • the oral administration was given once a day at 10 ml/kg for 30 days. .
  • Test data statistics Test data statistics were processed using SPSS 19.0 for windows software package.
  • the chlorogenic acid of the invention has a significant inhibitory effect on chemical liver injury, and the dosage is 50-140 mg/kg. Significant.

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Abstract

一种包含绿原酸晶型的制剂，其中所述晶型为正交晶系，空间群为P2 12 12 1，晶胞参数为a=7.7291（2）Å，b=10.9808（2）Å，c=36.5334（7）Å，α=β=γ=90.00°，Z=8，晶胞体积为3100.65（11）Å 3；所述制剂中的绿原酸晶型含量为10-5000mg/g。所述制剂有利于生物利用度，可用于制备治疗肿瘤、银屑病等免疫系统疾病、抗氧化、保肝利胆、心血管疾病、抗病毒的药物。

Description

一种包含绿原酸晶型的制剂及其用途 技术领域
本发明涉及一种包含绿原酸晶型的制剂及其用途，属于药物制剂领域。
背景技术
绿原酸，是由咖啡酸与奎尼酸生成的缩酚酸，是植物体在有氧呼吸过程中经莽草酸途径产生的一种苯丙素类化合物。
绿原酸具有广泛的生物活性，现代科学对绿原酸生物活性的研究已深入到食品、保健、医药和日用化工等多个领域。绿原酸是一种重要的生物活性物质，具有抗菌、抗病毒、增高白血球、保肝利胆、抗肿瘤、降血压、降血脂、清除自由基和兴奋中枢神经系统等作用。
然而，现有技术中的绿原酸制剂，大多以不定性的形式存在，其应用效果存在显著差异。
现有文献报道，绿原酸有多种晶型，但是对绿原酸晶型制剂的疗效效果对比研究未见报道。因为同一药物制剂的不同晶型常引起药物在外观、溶解度、熔点、密度等理化性质以及药物溶出度、生物有效性等显著差异，从而影响药物稳定性、生物利用度等疗效的发挥。
晶型对药物药效的影响是目前药学界比较关心的问题，同一药物在疗效上的差异，其原因除了因生产工艺上的不同而产生的质量差异外，另一个可能因素就是药物晶型的影响。药物的不同晶型，由于溶解度和溶出速率的不同，从而影响生物利用度，进而导致临床疗效的差异。文献报道西咪替丁存在A、B、C等多种晶型，仅A型最有效，而国产西咪替丁一般并非完全A型，从而影响了疗效。抗溃疡药法莫替丁有4种晶型，其熔点、红外光谱及理化性质差异明显，抑制胃酸分泌的活性B型大于A型。有些药物，其结晶状态反而不如非晶型疗效好，如无定型的新生霉素混悬剂。
发明内容
针对上述技术问题，本发明的目的在于提供一种包含特定绿原酸及其剂量的制剂。
本发明的上述目的是通过如下技术方案实现的：
一种包含绿原酸晶型的制剂，其中所述晶型为正交晶系，空间群为P212121，晶胞参数为
Figure PCTCN2015094869-appb-000001
α＝β＝γ＝90.00°，Z＝8，晶胞体积为
Figure PCTCN2015094869-appb-000002
所述制剂的临床剂量为每日1.0-20mg/kg。
在本发明中，根据不同剂型，所述制剂中的绿原酸晶型含量优选10-500mg/g。
在本发明中，所述制剂的剂型可以是本领域的常规剂型，包括但不限于口服制剂、注射剂，呼吸道给药制剂，皮肤给药制剂，粘膜给药制剂等。
申请人通过研究发现，控制绿原酸制剂中绿原酸的晶型，可以明显改善其使用效果。进一步的，申请人发现在所述注射制剂中，绿原酸晶型含量为10-60mg/kg时，所述制剂针对人脑胶质瘤、肝癌、肺癌、乳腺癌等肿瘤小鼠移植瘤有明显抑制作用具有更优的效果，优选为20-40mg/kg。根据动物与人的剂量换算关系，绿原酸冻干粉针剂的临床使用剂量保护：每日1.0-6.0mg/kg，优选为2-4mg/kg。
进一步的，申请人发现在所述口服制剂中，绿原酸晶型含量为20-200mg/kg时，所述制剂针对肝脏有明显保护作用，具有更优的效果，优选为50-140mg/kg。根据动物与人的剂量换算关系，绿原酸口服制剂的临床使用剂量范围：每日2mg-20mg/kg，优选为5-14mg/kg。
在本发明中，所述口服制剂及注射剂是由绿原酸或绿原酸和药学可接受的药用辅料制备而成。
在本发明中，所述绿原酸的晶型可以通过中国专利申请CN201410193699.8中记载的方式制备得到，所述冻干粉针剂也可以通过中国专利申请CN201310366945.0中记载的方式实施。
进一步的，在本发明中，所述冻干粉针剂包括如下组分：
绿原酸晶体  10-40重量份
抗氧化剂  1-5重量份
支架剂  60-100重量份
作为优选，上述冻干粉针剂中绿原酸晶体为30-40重量份；抗氧化剂优选2-5重量份；支架剂优选为80-90重量份。所述抗氧化剂为亚硫酸氢钠，支架剂为甘露醇。
在本发明的一个具体实例中，所述冻干粉针剂每1000个制剂单位包括如下组分：
绿原酸晶体  30g
抗氧化剂  2g
支架剂  80g
注射用水加至2000ml。
进一步的，本发明所述口服制剂包括如下组分：
绿原酸晶体  80-150重量份
填充剂  700-900重量份
粘合剂  20-220重量份。
作为优选，所述绿原酸晶体优选为80-100重量份；填充剂优选800-900重量份；粘合剂优选为100-220重量份。
在上述口服制剂中，所述填充剂包括但不限于淀粉、糖粉、糊精、乳糖、可压性淀粉、微晶纤维素、硫酸钙、磷酸氢钙或甘露醇中的一种或几种；粘合剂包括但不限于羧甲基纤维素钠、羟丙基纤维素、甲基纤维素、乙基纤维素、聚乙烯吡咯烷酮中的一种或几种。
在本发明的一个具体实例中，所述口服制剂每1000个制剂单位包括如下组分：
绿原酸晶体  100g
填充剂  800g
粘合剂  100g。
本发明的另一目的是提供一种上述制剂在制备治疗抗肿瘤、银屑病等免疫系统疾病、抗氧化、保肝利胆、心血管疾病、抗病毒等方面的药物中的应用，具体而言，包括但不限于小细胞肺癌、肝癌、乳腺癌、脑瘤、银屑病。
本发明的有益效果在于：本发明的绿原酸晶型制剂更有利于药物生物利用度等疗效的发挥，为绿原酸发挥疗效的优势晶型，且在治疗抗肿瘤、银屑病等免疫系统疾病、抗氧化、保肝利胆、心血管疾病、抗病毒等方面的药物中的应用发挥更为优良的临床疗效，并且本发明提供了该绿原酸晶型制剂用于临床的有效剂量，为该绿原酸晶型制剂用于临床的安全、有效提供依据。
附图说明
图1为本发明绿原酸与市售绿原酸对Lewis肺癌C57BL/6小鼠移植瘤抑瘤率的影响对比图。
图2为本发明绿原酸与市售绿原酸对H22肝癌KM小鼠移植瘤抑瘤率的影响对比图。
图3为本发明绿原酸与市售绿原酸对EMT-6乳腺癌BABLc小鼠移植瘤抑瘤率的影响对比图。
图4为本发明绿原酸与市售绿原酸对人脑胶质瘤小鼠抑瘤率的影响对比图。
具体实施方式
实施例1
冻干粉针剂的制备
按照1000个制剂单位(30mg/支)确定处方为：
绿原酸晶体  30g
抗氧化剂  2g
支架剂  80g
注射用水加至2000ml；
根据上述处方，称取原料，取注射用水依次加入抗氧化剂(如亚硫酸氢钠)、绿原酸晶体、支架剂(如甘露醇)，搅拌使充分溶解，pH控制在2-4；注射用水的温度为45-50℃，加入0.03％的活性炭，搅拌30分钟后过滤除去活性炭，再用0.22微米亲水性微孔滤膜精滤至滤液澄清，无菌灌装，冻干即得。
实施例2
口服制剂的制备
按照1000个制剂单位(100mg/片)确定处方为：
绿原酸晶体  100g
填充剂  800g
粘合剂  100g；
将绿原酸晶型加入适量的填充剂后及黏合剂后，过筛混合，制粒，整粒，加入适量的润滑剂，压片。
实施例3用于肺癌的治疗
动物 C57BL/6小鼠，体重13～20g，雌雄各半。
细胞系 Lewis细胞为小鼠肺癌细胞株，贴壁生长于含10％小牛血清，100U·ml-1青霉素、100μg·ml-1链霉素的RPMI-1640培养基中，于37℃、5％C02孵箱中培养，每2～3天换液传代一次。
细胞培养 细胞常规复苏，传代，将处于对数生长期Lewis细胞用0.25％胰蛋白酶消化脱壁后，终止消化，加入RPMI-1640培养液，吹打细胞制成悬液，1000rpm离心5分钟，洗涤2次，用台盼蓝染色法计数活细胞数。分别在小鼠左前肢腋窝皮下接种细胞悬液0.2ml·只-1(约含细胞数1×107个)，待肿瘤长大1cm×1cm×1cm时作为种鼠供实验用。无菌剥离种鼠肿瘤，用Hank′s液冲洗3次，清除血污、脂肪及坏死组织，将肿瘤剪切成1mm×1mm×1mm碎块，Hank's液冲洗2次，按比例加入生理盐水(1g:3ml)，再在玻璃匀浆器内研磨，经80-100目筛网过滤制成单细胞悬液，用台盼蓝染色法计数活细胞数。
(4)给药实验
将制备好的细胞悬液按0.2ml·只-1(约含细胞数1×107个)接种于C57BL/6小鼠左前腋窝下，并按体重随机分组，每组10只，分别为绿原酸给药剂量组100mg·kg-1、90mg·kg-1、80mg·kg-1、70mg·kg-1、60mg·kg-1、50mg·kg-1、40mg·kg-1、30mg·kg-1、20mg·kg-1、10mg·kg-1、5mg·kg-1、1mg·kg-1及阴性组(N.S生理盐水)。于接种后的第二日各组均腹腔注射(ip)给药，容量为0.2ml·10g-1，每日一次，连续给药12次。待阴性组平均瘤重大于1.0g(瘤体积约为0.5cm3)时停止实验，脱颈椎处死小鼠并称重，剥取肿瘤，计算抑瘤率。
抑瘤率％＝[1-(给药组平均瘤重/阴性组平均瘤重)]×100％。
(5)实验结果
由表1和图1可见，本发明绿原酸对Lewis肺癌C57BL/6小鼠移植瘤均有明显抑制作用，与阴性对照组比较，有统计学差异。且剂量组在10mg/kg-60mg/kg时抑瘤作用明显。
表1 本发明中绿原酸对Lewis肺癌C57BL/6小鼠移植瘤重量和抑瘤率的影响
Figure PCTCN2015094869-appb-000003
Figure PCTCN2015094869-appb-000004
Figure PCTCN2015094869-appb-000005
与阴性组比较*p＜0.05，**p＜0.01。
实施例4用于肝癌的治疗
(1)动物 昆明种小鼠，雌雄各半，体重16～27g。
细胞系 H22细胞株为小鼠肝癌细胞，悬浮生长于RPMI-1640培养基中，内含10％小牛血清，100U·ml-1青霉素和100μg·ml-1链霉素于37℃、5％C02孵箱中培养，每2～3天换液传代一次。
细胞培养 收获处于对数生长期的H22细胞，1000rpm离心5分钟，洗涤2次，用台盼蓝染色法计数活细胞数。尽快接种于KM小鼠腹腔内，7～10d后抽取腹水在小鼠左前肢腋窝皮下接种腹水细胞悬液0.2ml·只-1(约含细胞数2×107个)，待肿瘤长大1cm×1cm×1cm时作为种鼠供实验用。无菌剥离种鼠肿瘤，用Hank's液冲洗3次，清除血污、脂肪及坏死组织，将肿瘤剪切成1mm×1mm×1mm碎块，Hank's液冲洗2次，按比例加入生理盐水(1g:3ml)，再在玻璃匀浆器内研磨，经80-100目筛网过滤制成单细胞悬液，用台盼蓝染色法计数活细胞数。
(4)给药实验
将制备好的细胞悬液按0.2ml·只-1(约含细胞数1×106个)接种于KM小鼠左前腋窝下，并按体重随机分组，每组10只，分别为绿原酸给药剂量组100mg·kg-1、90mg·kg-1、80mg·kg- 1、70mg·kg-1、60mg·kg-1、50mg·kg-1、40mg·kg-1、30mg·kg-1、20mg·kg-1、10mg·kg-1、5mg·kg-1、1mg·kg-1及阴性组(N.S生理盐水)。于接种后的第二日开始静脉(iv)给药，容量为0.2ml·10g-1，每日一次。待阴性组平均瘤重大于1.0g(瘤体积约为0.5cm3)时停止实验，脱颈椎处死小鼠并称重，剥取肿瘤，计算抑瘤率。
(5)试验结果
由表2和图2可见，本发明绿原酸对H22肝癌KM小鼠移植瘤均有明显抑制作用，与阴性 对照组比较，有统计学差异。且剂量组在10mg/kg-60mg/kg时抑瘤作用明显。
表2 本发明绿原酸对H22肝癌KM小鼠移植瘤重量和抑瘤率的影响
Figure PCTCN2015094869-appb-000006
Figure PCTCN2015094869-appb-000007
与阴性组比较*p＜0.05，**p＜0.01。
实例5：BABLc小鼠移植瘤抑制作用的体内研究
实验材料
(1)动物 BABLc小鼠，♀，体重17～21g。
细胞系 EMT-6细胞株为小鼠乳腺癌细胞。贴壁生长于含10％小牛血清、1mmol/L谷胺酰胺、100U·ml-1青霉素和100μg·ml-1的RPMI-1640完全培养基中，于37℃、5％CO2孵箱中培养，每2～3天换液传代一次。
细胞培养 细胞常规复苏，传代，将处于对数生长期的EMT-6细胞用0.25％的胰蛋白酶消化脱壁后，终止消化，加入RPMI-1640培养液，吹打细胞制成悬液，1000rpm离心5分钟，洗涤2次，用台盼蓝染色法计数活细胞数。尽快分别在小鼠左前肢腋窝皮下接种细胞悬液0.2ml·只-1(约含细胞数1×107个)，待肿瘤长大1cm×1cm×1cm时作为种鼠供实验用。无菌剥离种鼠肿瘤，用Hank's液冲洗3次，清除血污、脂肪及坏死组织，将肿瘤剪切成1mm×1mm×1mm碎块，Hank's液冲洗2次，按比例加入生理盐水(1g:3ml)，再在玻璃匀浆器内研磨，经80-100目筛网过滤制成单细胞悬液，用台盼蓝染色法计数活细胞数。
实验方法
将制备好的细胞悬液按0.2ml·只-1(含细胞数约1×106个)接种于70只BABLc小鼠左前腋窝下，并按体重随机分组，每组10只，分别为绿原酸给药剂量组100mg·kg-1、90mg·kg-1、80mg·kg-1、70mg·kg-1、60mg·kg-1、50mg·kg-1、40mg·kg-1、30mg·kg-1、20mg·kg-1、10mg·kg-1、5mg·kg-1、1mg·kg-1及阴性组(N.S生理盐水)。于接种后的第二日各组均肌肉注射(ip)给药，容量为0.2ml·10g-1，每日一次，连续给药9天。待阴性组平均瘤重大于1.0g(瘤体积约为0.5cm3)时停止实验，脱颈椎处死小鼠并称重，剥取肿瘤，计算抑瘤率。
抑瘤率％＝[1-(给药组平均瘤重/阴性组平均瘤重)]×100％。
实验结果
绿原酸对EMT-6乳腺癌BABLc小鼠移植瘤重量和抑瘤率影响见表3和图3。
表3 绿原酸对EMT-6乳腺癌BABLc小鼠移植瘤重量和抑瘤率的影响
Figure PCTCN2015094869-appb-000008
组别 剂量(mg·kg-1) 动物数(n) 瘤重(g) 抑瘤率(％)
本发明绿原酸 100 10 0.995±0.0256 25.80
本发明绿原酸 90 10 0.859±0.0754 35.94
本发明绿原酸 80 10 0.789±0.087 41.16
本发明绿原酸 70 10 0.698±0.112*** 47.95
本发明绿原酸 60 10 0.681±0.156*** 49.22
本发明绿原酸 50 10 0.529±0.168*** 60.55
本发明绿原酸 40 10 0.451±0.097*** 66.37
本发明绿原酸 30 10 0.429±0.168*** 68.01
本发明绿原酸 20 10 0.403±0.101*** 69.95
本发明绿原酸 10 10 0.690±0.188*** 48.55
本发明绿原酸 5 10 0.789±0.087 41.16
本发明绿原酸 1 10 0.987±0.154 26.40
市购绿原酸 100 10 1.312±0.087 2.16
市购绿原酸 90 10 1.298±0.423 3.21
市购绿原酸 80 10 1.279±0.143 4.62
市购绿原酸 70 10 1.254±0.514 6.49
市购绿原酸 60 10 1.009±0.123 24.76
市购绿原酸 50 10 1.115±0.201 16.85
市购绿原酸 40 10 1.125±0.125 16.11
市购绿原酸 30 10 1.185±0.247 11.63
市购绿原酸 20 10 1.201±0.546 10.44
市购绿原酸 10 10 1.274±0.854 5.00
市购绿原酸 5 10 1.285±0.324 4.18
市购绿原酸 1 10 1.299±0.253 3.13
阴性组 N.S 10 1.341±0.299 -
。
与阴性组比较***p＜0.001。
由表3和图3可见，本发明绿原酸对EMT-6乳腺癌BABLc小鼠移植瘤均有明显抑制作用，与阴性对照组比较，有统计学差异。且剂量组在10mg/kg-60mg/kg时抑瘤作用明显。
实例6：绿原酸对移植性脑胶质瘤抑制作用的研究
1.材料
人脑胶质瘤细胞株，由四川大学华西医院卫生部移植工程和移植免疫重点实验室提供。本发明绿原酸，由四川九章生物科技有限公司提供。市购绿原酸。
方法
(1)造模 小鼠随机分组，每组10只。将脑胶质母细胞瘤细胞株配比合适的稀释液，分别接种于小鼠左颞叶皮质内。
给药干预 接种24h后，分别对各组小鼠进行腹腔注射给药。分别为绿原酸100mg·kg-1、90mg·kg-1、80mg·kg-1、70mg·kg-1、60mg·kg-1、50mg·kg-1、40mg·kg-1、30mg·kg-1、20mg·kg-1、10mg·kg-1、5mg·kg-1、1mg·kg-1剂量组，阴性对照组给相同体积的生理盐水， 连续给药15d。
测抑瘤率 最后一日停止给药并且将所有小鼠处死，解剖小鼠,取肿瘤组织并且称重。抑制率＝(对照组平均瘤重-治疗组平均瘤重)/瘤重对照组平均瘤重×100％。
统计学方法 数据分析采用统计程序软件包(SPSS 13.0for Windows)进行方差分析，以P＜0.05为差异有显著性。
结果
经过分析结果发现，给药组相对于空白组其抑瘤率有显著提升的态势，证明绿原酸有抗肿瘤生长之功效，其抑瘤效果以中剂量组最为明显，高剂量组次之，具体数据请见表4。
表4 绿原酸对人脑胶质瘤小鼠瘤重及抑瘤率的影响
Figure PCTCN2015094869-appb-000009
Figure PCTCN2015094869-appb-000010
Figure PCTCN2015094869-appb-000011
由表4和图4可见，本发明绿原酸对人脑胶质瘤小鼠移植瘤有明显抑制作用，与阴性对照组比较，有统计学差异。且剂量组在10mg/kg-60mg/kg时抑瘤作用明显。
实例7：绿原酸对化学性肝损伤(酒精性脂肪肝大鼠模型)的保护作用
1.材料和方法
药物：本发明绿原酸由四川九章生物科技有限公司提供。市购绿原酸。
动物：雌性SD大鼠，体重为156～193g，SPF级。
试验分组及剂量设计：将雌性SD大鼠随机分组，每组10只动物，试验设20mg/kg、35mg/kg、50mg/kg、70mg/kg、85mg/kg、100mg/kg、120mg/kg、140mg/kg、160mg/kg、180mg/kg、200mg/kg剂量组，另设阴性对照组和50％乙醇模型对照组。用乙醇(分析纯)造成肝损伤模型。
试验方法：采用酒精肝损伤模型法，绿原酸组灌胃给予不同剂量的受试药物，阴性对照组和模型对照组给予生理盐水，按10ml/kg每天经口灌胃一次，连续给予30天。
2.检测指标
大鼠血清ALT、AST、TG的测定，肝匀浆SOD、MDA、还原型谷胱甘肽(GSH)的测定。
试验数据统计：试验数据统计采用SPSS 19.0for windows软件包处理。
实验结果
绿原酸对大鼠肝匀浆中SOD、MDA、还原型GSH和血清ALT、AST、TG含量的影响见表5。
由表5可见，模型对照组大鼠肝匀浆中的MDA、GSH、TG含量与阴性对照组比较，MDA、TG含量显著升高(P＜0.01)，还原型GSH含量显著降低(P＜0.01)，表明该模型是成功的，实验系统可靠。而本发明绿原酸组均能不同程度的降低大鼠血清ALT、AST、TG、MDA含量，提高SOD、GSH水平；且剂量组在50mg/kg-140mg/kg时，具有极显著的统计学意义。
因此本发明绿原酸对化学性肝损伤有明显抑制作用，且使用剂量50-140mg/kg作用最 显著。
表5 绿原酸对大鼠肝匀浆中SOD、MDA、还原型GSH和血清ALT、AST、TG含量的影响
Figure PCTCN2015094869-appb-000012
Figure PCTCN2015094869-appb-000013
Figure PCTCN2015094869-appb-000014
注：与阴性对照组比较：**P<0.01，*P<0.05；与模型对照组比较:##P<0.01，#P<0.05。

Claims (10)

  1. 一种包含绿原酸晶型的制剂，其中所述绿原酸晶型为正交晶系，空间群为P212121，晶胞参数为
    Figure PCTCN2015094869-appb-100001
    α＝β＝γ＝90.00°，Z＝8，晶胞体积为
    Figure PCTCN2015094869-appb-100002
    所述制剂的临床剂量为每日1.0-20mg/kg。
  2. 如权利要求1所述的制剂，其中制剂为口服制剂或注射剂，所述注射剂优选为冻干粉针剂。
  3. 如权利要求2所述的制剂，其中所述注射制剂临床剂量为每日1.0-6.0mg/kg，优选为2-4mg/kg。
  4. 如权利要求2所述的制剂，其中所述口服制剂的临床剂量每日2mg-20mg/kg，优选为5-14mg/kg。
  5. 如权利要求2或3所述的制剂，其中所述注射剂包括如下组分：
    绿原酸晶体  10-40重量份
    抗氧化剂  1-5重量份
    支架剂  60-100重量份；
    优选绿原酸晶体为30-40重量份；抗氧化剂优选2-5重量份；支架剂优选为80-90重量份；优选所述抗氧化剂为亚硫酸氢钠，支架剂为甘露醇。
  6. 如权利要求5所述的制剂，其中所述注射剂每1000制剂单位包括如下组分：
    绿原酸晶体  30g
    抗氧化剂  2g
    支架剂  80g
    注射用水加至2000ml。
  7. 如权利要求2或3所述的制剂，其中所述口服制剂包括如下组分：
    绿原酸晶体80-150重量份
    填充剂700-900重量份
    粘合剂20-220重量份；
    优选所述绿原酸晶体优选为80-100重量份；填充剂优选800-900重量份；粘合剂优选为100-220重量份；优选所述填充剂选自淀粉、糖粉、糊精、乳糖、可压性淀粉、微晶纤维素、硫酸钙、磷酸氢钙或甘露醇中的一种或几种；所述粘合剂选自羧甲基纤维素钠、羟丙基纤维素、甲基纤维素、乙基纤维素、聚乙烯吡咯烷酮中的一种或几种。
  8. 如权利要求7所述的制剂，其中所述口服制剂每1000个制剂单位包括如下组分：
    绿原酸晶体  100g
    填充剂  800g
    粘合剂  100g。
  9. 权利要求1-8任一项所述制剂在制备治疗免疫系统疾病、抗氧化、保肝利胆、心血管疾病、抗病毒等的药物中的应用。
  10. 如权利要求9所述的应用，所述免疫系统疾病为肿瘤、银屑病等；所述肿瘤包括但不限于小细胞肺癌、肝癌、乳腺癌或脑瘤。
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