TWI248431B - Chlorogenic acid, isochlorogenic acid composition, and pharmacology use thereof - Google Patents

Abstract

The present invention disclosed a composition containing chlorogenic acid and isochlorogenic acid extracted from natural plant. Medical tests indicated that the invented composition has conspicuous anti-SARS-viral activities. Chlorogenic acid and isochlorogenic acid with active and Vitamin C are used as main active ingredients, added with adjuvant, and an auxiliary agent or carrier to prepare different types of compositions. The invented compositions can be applied in the treatment and prevention of diseases caused by SARS virus.

Description

1248431 玖, the invention description: [Technical field of the invention] The present invention relates to a biologically active compound, more specifically to a composition of chlorogenic acid, isochlorogenic acid and its anti-SARS virus activity in the preparation of prevention and treatment of atypical Application in pneumonia drugs. [Prior Art] Chlorogenic acid, isochlorogenic acid, and xanthine substances are present in various plants, such as honeysuckle, coffee, Eucommia ulmoides, and Artemisia scoparia. Gold, flower, also known as double flower, silver flower, honeysuckle flower, is the perennial entwined woody vine of the honeysuckle family. China is distributed throughout the north and south. At the beginning of summer, when the flower buds contain t, they are picked, darkened or dried, used or fried. Sexual cold, sweet, into the lungs, stomach, large intestine. Function: clearing away heat and detoxifying, dispersing wind and heat, cooling blood and stopping disease. Chinese medicine believes that honeysuckle is sweet and cold, and its gas is fragrant. It can not only clear away heat and detoxification, but also dispel the wind and heat. It can be revealed through clearing, the heat of external treatment, the early stage of warm disease, and the swelling of the disease. fun. I often drink tea in the golden and raw licorice, but I can use the summer heat to resist the temperature, ', month', ', thirst quenching' to reduce the gold and silver carp, Ophiopogon japonicus, platycodon, ebony, and cannon. Chronic pharyngitis. Chlorogenic acid (10) (10) genic acid), synonym: if non-transacid, is a styrene-flavored quinine _ class compound towel most «the day of the seam -", mainly stored - some of the traditional scurvy Among the flowers, 'the ingredients of these caps.' The structural formula of chlorogenic acid is as follows (1):

HO COOH

HO COOH, the structural formula of iS〇Chl〇rogenic acid may be as follows (9) or (ιι)

S=CHCOO〆 Π

OOCCH=c (II)

1248431

It can cause asthma, New Zealand, etc., but the eclipse can stop bleeding, increase blood cells, have blood test and "time effect. Chlorogenic acid toxicity = young rats oral LD5 () is greater than ig / Kg intraperitoneal injection is greater than 〇 · 25g/Kg. ” [Abstract] The object of the present invention is to provide a new medical use of chlorogenic acid and isochlorogenic acid, and the anti-SARS virus activity of the special star can be used for preparing a medicament for preventing and treating SARS virus diseases. Further application of the present invention is to provide a composition comprising chlorogenic acid or isochlorin which is active against SARS virus as a main component to prepare a medicament for treating or preventing diseases caused by prion. A further object is to formulate the composition into a suitable dosage form. μ ° The chlorogenic acid of the invention is dehydrated from the plant honeysuckle twice, and the decoction is concentrated under reduced pressure to -^, and the lime milk suspension is adjusted to pH, and the mixture is separated by centrifugation. Once the carcass, 1 mouth, 酉 酉 研 匀 匀 匀 匀 匀 匀 匀 匀 匀 匀 匀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀 稀Pure acid.

(chlorogenic acid structure) S4. 5.2 0 1248431 Chlorogenic acid has been proved to have significant anti-SARS virus activity by pharmacological test. Vero-E6 cells are used as viral host cells (susceptible cells) for the determination of chlorogenic acid. The protective index of virus-infected cells is the cell degeneration reaction and the protection rate of infected cells. The virus strain was tested by the BJ-01 ''s sample of 7 dilutions' and proved to have significant anti-mrs virus activity. According to the chlorogenic acid and isochlorogenic acid, the anti-SARS virus activity is combined with the medical use characteristics of vitamin strontium to prepare a new composition, and the vitamin C, chlorogenic acid or isochlorogenic acid: vitamin C weight ratio is 10 : 1-1: 10. Further, adjuvants are added as needed to prepare compositions of different dosage forms. The invention comprises an effective amount of chlorogenic acid, isochlorogenic acid and a pharmaceutically acceptable excipient, carrier or diluent to prepare different pharmaceutical dosage forms, which can chlorogenic acid, isochlorogenic acid and vitamin strontium and oral administration Typical pharmaceutical carriers and excipients in the preparation are formulated as oral preparations, and pharmaceutical carriers and excipients include lactose; powders and derivatives thereof such as corn starch, sodium carboxymethyl starch, etc.; cellulose derivatives such as methyl And ethyl cellulose; gelatin; inorganic compounds such as light magnesium oxide, talc, etc.; oils such as vegetable oils, sesame oils, etc.; inclusion agents such as /3-cyclodextrin; and glycols such as polyethylene glycol. The oral preparation can be used as a tablet, a capsule, an emulsion, a solution, etc., and can also be applied to a solid dispersion tablet, a lipid system targeting preparation, and a controlled release preparation. The present invention comprises an effective amount of chlorogenic acid, isochlorogenic acid and a pharmaceutically acceptable shaped carrier or diluent formulated into an injection comprising isotonic saline, 5% dextrose in water, water for injection, tea oil, soybean oil, sesame oil, Ethanol, glycerin, propylene glycol, polyethylene glycol, benzyl benzoate; TWEEN8〇, egg fat, sodium deoxycholate, egg yolk phospholipid, polyvinylpyrrolidone; acetic acid, sodium acetate, citric acid and sodium salt, lactic acid, tartaric acid And sodium salt, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium hydrogencarbonate and sodium carbonate; gelatin, sodium carboxymethylcellulose, pectin, sorbitol solution; creatinine, glycine; sodium sulfite, sodium bisulfite, Jiaoya Sodium phosphate, sodium thiosulfate, benzoyl alcohol, butyl p-hydroxybenzoate and ethyl phenol, chlorobutanol; lidocaine; cephalin, soybean phospholipid, etc., the typical form of injection preparation is powder injection, mixed Suspension injections, etc. In the tablet, the composition can be made into a brewing agent with a disintegrant composed of a mixture of sodium hydrogencarbonate and tartaric acid or citric acid, and when it is in contact with water, carbon dioxide gas is generated to rapidly disintegrate the tablet. 1248431—The compound can also be made into an aerosol, for example, an aerosol formed by using a polyvinyl chloride paste resin, dibutyl phthalate, dioctyl benzoate, calcium stearate, and stearic acid. Agent. [Embodiment] Take honeysuckle water decoction 2 times, decoction concentrated to 1: 2 under reduced pressure, add 20% lime milk suspension to pH 10 or so, centrifuge to separate the precipitate, and centrifuge the precipitate with 2 times the amount of ethanol. Under stirring, 5 〇 % thiol text > gluten solution was adjusted to pH 3, stirred well, and the solid matter was removed by centrifugation. The filtrate was adjusted to Qiu 6 with 2NNa〇H, ethanol was recovered, and dried under reduced pressure to obtain a crude extract. The crude extract was dissolved in a small amount of water, separated by polyamine column chromatography, eluted with a gradient of ethanol-water, and the eluate was detected by a thin layer of polyamine. The chlorogenic acid fraction was combined and dried under reduced pressure. Recrystallization from water gives pure chlorogenic acid. Chlorogenic acid anti-SARS virus activity results / test project: screening of chlorogenic acid anti-SARS virus activity (virus-cell standard) Test principle: using Vero-E6 cells as viral host cells (susceptible cells), test sample pairs The protective effect of virus-infected cells, the detection index is the cell degeneration reaction (CPE) (Cyt〇pathic Effect) and the protection rate of infected cells. Test materials and methods: chlorogenic acid strain: BJ-01 Sample treatment: The sample is dissolved in the culture solution or DMS0, and is formulated into a suitable initial concentration, 5 times dilution, 7 dilutions each. Test method: Vero-E6 cells were seeded in a 96-well culture plate, cultured at 37 ° C in a 5% CO 2 incubator, and SARS virus and samples of different dilution concentrations were added, respectively, and virus control, cell control and sample control were set. The results of daily microscopic observations were recorded, CPE was recorded, and the value of (8) was determined by neutral red staining. The calculation and evaluation of the anti-SARS virus activity of the samples were carried out with reference to the control. Test result - concentration Ug / ml) CFE infection fine - m - - one - 1248431 ο 8163206 2040..1000 00.0. 457831 567332 2.2.2.2.2.1. Know the green like magic: "" represents the cell without lesions; Indicates the degree of cytopathic effect, <25% +, 25% to 50% ++, 50% to 75 +++, >75%++++.消杂游#·· By comparing the virus control, cell control and sample check (8) values, calculate the protective activity of the sample on virus-infected cells, the protection rate is W rate, and it is preliminarily considered that the sample has certain protective activity against virus-infected cells. . * 揉 续 · · · If the cytotoxicity of the sample and the cell control ratio do not do cpE evaluation and protection rate calculation. The above results show that the compound has an anti-SARS virus active side. The LD5G of chlorogenic acid is greater than lg/Kg, and the intraperitoneal injection is greater than 0 25g/Kg, indicating less toxicity. In the first embodiment, the following dosages of the pharmaceutical composition and the additional agent are selected: (compound··vitamin=1: iw/w) 400 g of active compound, % vitamin C, pre-gelatinized powder of magnesium stearate, 400 g of vitamin C powder 20 grams of 60 grams of 120 grams of the appropriate amount of TSbo granules Nanxun music composition and additives are evenly mixed, with 10% pre-gelatinized powdered polyglycol as a dry 'w rope 1L 'for dry' plus the right amount of hard fat duck mixed, pressed Bonding agent. Example 2 of the pharmaceutical composition and additive: (Compound: Wisconsin = 1.9) Active compound 80 g f Wang I i.bu 10 1248431 Vitamin C 720 g of vitamin C 20 g of micronized gelatin 48 g of ethanol amount of stearic acid Magnesium sulphate - prepared by mixing the above-mentioned drug mixture with hydroxypropyl cellulose and micro-powder gel, using a suitable amount of 7〇% ethanol as wet granules, wet granulation, screening through 40 mesh into granules, drying, adding hard Magnesium citrate, mixed, filled into hard capsules. The third embodiment uses the following dosages of the pharmaceutical composition and the additive: (compound: vitamin = 2:1) active compound 800 g vitamin C 400 g sucrose 40 g preservative proper amount of distilled water ~ ^ 20000 ^ 1 ---- take distilled water An appropriate amount of hydrazine is added to the above pharmaceutical composition, stirred while heating to dissolve, and filtered. Further, sucrose and a preservative are dissolved by heating in distilled water, and the mixture is slowly added to the above solution under stirring, and distilled water is added to the whole amount, mixed, refrigerated, and sterilized to obtain an oral solution. Example 4 The following dosages of pharmaceutical compositions and additives are used: (compound: vitamin = 6:1) active compound 240 g of vitamin c 4 g

Tween80 Appropriate amount 11 1248431 94. δ. 2 适 Injectable water amount - Made: 45000 ml Take appropriate amount of water for injection, add the above pharmaceutical composition, heat, stir to fully dissolve, add 4% Tween80, stir well: evenly, filter, add Adjust the pH of hydrochloric acid to 6.8, add water for injection to a sufficient amount, hollow fiber ultrafiltration, fine filtration, encapsulation, sterilization, and get the injection. [Simple description of the map]

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Claims (1)

1248431 7. The composition according to claim 6, which has anti-SARS virus activity, which is in the form of a powder injection or a suspension injection. 8. A composition having anti-SARS virus activity according to claim 1, wherein the ingot is made of sodium hydrogencarbonate, tartaric acid or citric acid. 9. The composition of claim 1 having anti-SARS virus activity, which is formulated with a gas-gas vinyl paste resin, dibutyl phthalate, dioctyl phthalate, calcium stearate or zinc stearate. Made into an aerosol. 10. A composition having anti-SARS virus activity according to claim 1 for use in the preparation of a medicament for preventing or treating a disease caused by a SARS virus. 14