CN113041352B - Chlorogenic acid-containing pharmaceutical composition, preparation method and application thereof - Google Patents
Chlorogenic acid-containing pharmaceutical composition, preparation method and application thereof Download PDFInfo
- Publication number
- CN113041352B CN113041352B CN202110265517.3A CN202110265517A CN113041352B CN 113041352 B CN113041352 B CN 113041352B CN 202110265517 A CN202110265517 A CN 202110265517A CN 113041352 B CN113041352 B CN 113041352B
- Authority
- CN
- China
- Prior art keywords
- chlorogenic acid
- pharmaceutical composition
- capc
- group
- phospholipid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pulmonology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Emergency Medicine (AREA)
- Organic Chemistry (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a pharmaceutical composition containing chlorogenic acid and a preparation method and application thereof, wherein the preparation method comprises the following steps: (1) Weighing phospholipid and the first part of chlorogenic acid, adding ethanol water solution, stirring, recovering ethanol under reduced pressure, drying the solid under reduced pressure, pulverizing, and sieving to obtain chlorogenic acid-phospholipid complex; (2) Weighing a second part of chlorogenic acid and chlorogenic acid phospholipid complex, mixing, and uniformly stirring to obtain the pharmaceutical composition containing chlorogenic acid. The composition has high oral bioavailability, improves the oral pharmacokinetic parameters of the chlorogenic acid, and can remarkably improve the survival effect of the chlorogenic acid on lung injury, inhibit weight loss, inhibit the effect of lung tissue inflammatory factors, resist inflammatory storm, resist respiratory failure caused by the lung injury and quickly recover the respiratory function of the lung, thereby improving the curative effect of treating and/or preventing acute lung injury. In the composition, the chlorogenic acid and chlorogenic acid phospholipid complex shows excellent synergistic effect.
Description
Technical Field
The invention relates to the field of medicines, in particular to a chlorogenic acid-containing pharmaceutical composition, a preparation method and application thereof.
Background
Acute Lung Injury (ALI) refers to Acute hypoxic respiratory insufficiency or respiratory failure caused by diffuse interstitial pulmonary and alveolar edema due to Injury of pulmonary capillary endothelial cells and alveolar epithelial cells in the process of non-cardiogenic diseases such as infection, inhalation Injury (toxic gas, acid reflux, drowning and the like), mechanical Injury, iatrogenic Lung Injury of radiotherapy and chemotherapy, severe pancreatitis, sepsis or severe non-chest trauma and the like, and the ALI of a patient greatly harms human health. ALI is pathophysiologically characterized by decreased lung volume, decreased lung compliance, a disturbed ratio of ventilation/blood flow, clinically manifested as progressive hypoxemia and respiratory distress, and pulmonary imaging manifested as non-uniform exudative lesions, which progress to a severe stage (oxygenation index < 200) known as Acute Respiratory Distress Syndrome (ARDS). Although the pathogenesis of ALI/ARDS has not been fully elucidated, studies have demonstrated that the major pathogenesis of ALI/ARDS, for whatever reason, is associated with inflammatory storms. Currently, clinical treatments for ALI/ARDS can be divided into supportive care and drug intervention. Mechanical ventilation is a commonly used clinical supportive treatment, and although effective in improving patient prognosis, patient mortality remains as high as 35% -45%. In addition, there are many problems with the clinical use of ALI/ARDS for drug therapy. The beta-receptor agonist salbutamol was originally thought to reduce extravascular pulmonary edema and respiratory pressure, but clinical studies have shown that salbutamol is poorly tolerated by intravenous injections and is detrimental to patient prognosis. Other clinical effects such as aspirin, corticosteroids, antibiotics and the like are not expected. At present, although a great deal of research is carried out on the main pathogenesis of ALI/ARDS at home and abroad, no effective therapeutic drug is found, symptomatic treatment is mainly used for treatment, and the fatality rate of patients still remains high. Therefore, the exploration of new drugs and the action mechanism thereof have important significance for the clinical treatment of ALI/ARDS.
Chlorogenic acid (CGA) is widely found in medicinal plants, mainly in plants of Caprifoliaceae, and Eucommiaceae, such as flos Lonicerae and Eucommiae cortex. Chlorogenic acid has solubility of 4% in water at 25 deg.C, and higher solubility in hot water, and is soluble in ethanol and acetone, slightly soluble in ethyl acetate, and insoluble in chloroform and diethyl ether.
At present, research shows that chlorogenic acid has the curative effect of preventing and treating lung injury, can inhibit the increase of the activity of lung tissue myeloperoxidase caused by lipopolysaccharide and the expression of polymorphonuclear leukocytes in alveolar lavage fluid, and simultaneously obviously reduces the activity of inducible nitric oxide synthase in lung tissue; in addition, chlorogenic acid also has obvious protective effect on lung injury caused by hydrogen peroxide and cobra venom factors. Although chlorogenic acid has good pharmacological action, chlorogenic acid has poor oral bioavailability and unsatisfactory pharmacokinetic parameters, and the curative effect is seriously influenced. Therefore, the key of the chlorogenic acid in drug property is that the oral bioavailability of the chlorogenic acid is improved, the oral pharmacokinetic parameters of the chlorogenic acid are improved, and the pharmacological pharmacodynamic action of the chlorogenic acid is improved.
Disclosure of Invention
The invention aims to overcome the defects of poor oral bioavailability, unsatisfactory pharmacokinetic parameters and low curative effect of chlorogenic acid and provides a pharmaceutical composition containing the chlorogenic acid.
The second purpose of the invention is to provide a preparation method of a chlorogenic acid-containing pharmaceutical composition.
The third purpose of the invention is to provide the application of the pharmaceutical composition containing chlorogenic acid in preparing the medicines for preventing and treating acute lung injury.
The technical scheme of the invention is summarized as follows:
the preparation method of the chlorogenic acid-containing pharmaceutical composition comprises the following steps:
(1) Weighing phospholipid and first chlorogenic acid, placing in a container, adding ethanol water solution, stirring at 20-35 deg.C for 1-2 hr, recovering ethanol under reduced pressure, drying the solid under reduced pressure, pulverizing, and sieving to obtain chlorogenic acid phospholipid complex;
(2) Weighing a second part of chlorogenic acid and the chlorogenic acid phospholipid complex obtained in the step (1), mixing or mixing according to an equivalent incremental method, and uniformly stirring to obtain the pharmaceutical composition containing chlorogenic acid.
Preferably, the mass ratio of the phospholipid to the first chlorogenic acid to the ethanol aqueous solution is (1-10): (1-10): (100-4000).
Preferably, the mass ratio of the second part of chlorogenic acid to the chlorogenic acid phospholipid complex obtained in the step (1) is (1-10): (1-20).
The phospholipid is at least one of soybean phospholipid, egg yolk lecithin, dimyristoyl lecithin, dipalmitoyl lecithin, distearoyl lecithin, dioleoyl lecithin, 1-myristoyl-2-palmitoyl lecithin, 1-stearoyl-2-myristoyl lecithin, myristoyl lysolecithin, palmitoyl lysolecithin, stearoyl lysolecithin, dilauroyl phosphatidylglycerol, dimyristoyl phosphatidylglycerol, 1,2-palmitoyl phosphatidylglycerol, distearoyl phosphatidylglycerol, dimyristoyl phosphatidic acid, dipalmitoyl phosphatidic acid, distearoyl phosphatidic acid, dimyristoyl phosphatidylethanolamine, dioleoyl phosphatidylethanolamine, dipalmitoyl phosphatidylethanolamine, distearoyl ethanolamine, 1-palmitoyl-2-oleoyl phosphatidylethanolamine, dimyristoyl phosphatidylserine, dioleoyl phosphatidylserine and distearoyl phosphatidylserine.
The volume concentration of the ethanol water solution is 70% or more;
preferably, the screening is through a third to a seventh screen.
Preferably, the sieve is a No. five sieve.
The pharmaceutical composition containing chlorogenic acid prepared by the method.
The application of the chlorogenic acid-containing pharmaceutical composition in preparing medicines for preventing and treating acute lung injury is provided.
The invention has the advantages that:
the pharmaceutical composition containing the chlorogenic acid has high oral bioavailability, improves the oral pharmacokinetic parameters of the raw material chlorogenic acid, and can remarkably improve the survival effect of the chlorogenic acid on lung injury, the weight reduction inhibition effect of the chlorogenic acid, the lung tissue inflammation factor inhibition effect of the chlorogenic acid, the anti-inflammatory storm effect of the chlorogenic acid, the respiratory failure effect of the chlorogenic acid on lung injury and the rapid lung respiratory function recovery effect of the chlorogenic acid, thereby improving the curative effect of the chlorogenic acid on treating and/or preventing acute lung injury. In the chlorogenic acid-containing pharmaceutical composition provided by the invention, the chlorogenic acid and chlorogenic acid phospholipid complex shows excellent synergistic effect.
Drawings
FIG. 1 Mean plasma concentration-time curve (Mean + -SD) of chlorogenic acid (CGA) in rat plasma after administration of chlorogenic acid (CGA), chlorogenic Acid Phospholipid Complex (CAPC), pharmaceutical composition containing chlorogenic acid of the present invention (CA-CAPC);
FIG. 2 is a schematic representation of the effect of chlorogenic acid (CGA), chlorogenic Acid Phospholipid Complex (CAPC), a chlorogenic acid containing pharmaceutical composition of the present invention (CA-CAPC) on the survival of mice;
FIG. 3 is a schematic diagram showing the effect of chlorogenic acid (CGA), chlorogenic Acid Phospholipid Complex (CAPC) and the pharmaceutical composition containing chlorogenic acid (CA-CAPC) of the present invention on the weight of mice (A: weight of mice; B: weight loss of mice)
Note: # p < 0.05vs blank group; # p < 0.01vs blank group; # p < 0.001vs blank group;
* p < 0.05vs model group; * P < 0.01vs model group; * P < 0.01vs model group;
DELTA p < 0.05vs CGA group; delta < delta > p < 0.01vs CGA group; delta deltap is less than the 0.001vs CGA group;
the fourth p is less than 0.05vs CAPC group; the four-star p is in the CAPC group of 0.01 vs; it is called the giro 0.001vs CAPC group.
FIG. 4 is a study of the effect of chlorogenic acid (CGA), chlorogenic Acid Phospholipid Complex (CAPC), and pharmaceutical composition containing chlorogenic acid (CA-CAPC) on LPS-induced lung tissue inflammatory factor expression in mice; (wherein A: IL-6, B TNF- α; C: IL-1 β): # p < 0.05vs blank group; # p < 0.01vs blank group; the # p < 0.001vs blank group;
* p < 0.05vs model group; * P < 0.01vs model group; * P < 0.01vs model group;
DELTA p < 0.05vs CGA group; delta < delta > p < 0.01vs CGA group; delta deltap is less than the 0.001vs CGA group;
the fourth p is less than 0.05vs CAPC group; the four-star p is in the CAPC group of 0.01 vs; it is a girit < 0.001vs CAPC group.
FIG. 5 study on the effect of chlorogenic acid (CGA), chlorogenic Acid Phospholipid Complex (CAPC), and pharmaceutical composition containing chlorogenic acid (CA-CAPC) on lung respiratory function of mice; ( A: tidal volume-time curve; b: tidal volume; c: minute ventilation versus time curve; d: ventilation per minute )
Note: # p < 0.05vs blank group; # p < 0.01vs blank group; # p < 0.001vs blank group;
* p < 0.05vs model group; * P < 0.01vs model group; * Left < 0.01vs model group;
DELTA p < 0.05vs CGA group; delta < delta > p < 0.01vs CGA group; delta deltap is less than the 0.001vs CGA group;
the fourth p is less than 0.05vs CAPC group; the four-star p is in the CAPC group of 0.01 vs; it is a girit < 0.001vs CAPC group.
FIG. 6 comparative analysis of chlorogenic acid (CGA), chlorogenic Acid Phospholipid Complex (CAPC), and chlorogenic acid-containing pharmaceutical composition (CA-CAPC) on lung respiratory function of mice by a pre-assay/model group; ( A: relative tidal volume change values; b: relative change in minute ventilation )
Note: DELTA p < 0.05vs CGA group; Δ p < 0.01vs CGA group; delta deltap is less than the 0.001vs CGA group;
the fourth p is less than 0.05vs CAPC group; the four-turn p is in the 0.01vs CAPC group; it is a girit < 0.001vs CAPC group.
Detailed Description
The first chlorogenic acid and the second chlorogenic acid are the same and are extracted from plants or chemically synthesized as raw material medicines, the chlorogenic acid adopted in the embodiments of the invention is obtained from Shanghai Allantin Biotechnology Co., ltd, the purity is more than 98%, and chlorogenic acid sold by other enterprises can also be used in the invention.
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The present invention will be further specifically illustrated by the following examples for better understanding, but the present invention is not to be construed as being limited thereto, and certain insubstantial modifications and adaptations of the invention by those skilled in the art based on the foregoing disclosure are intended to be included within the scope of the invention.
The present invention will be further illustrated by the following specific examples.
Example 1
The preparation method of the chlorogenic acid-containing pharmaceutical composition comprises the following steps:
(1) Weighing soybean phospholipid and a first part of chlorogenic acid, placing into a container, adding 75 vol% ethanol water solution, stirring at 30 deg.C for 2h, recovering ethanol under reduced pressure (50 deg.C and-0.08 Mpa), drying the solid under reduced pressure, pulverizing the dried solid, and sieving with a five-mesh sieve to obtain chlorogenic acid phospholipid complex;
(2) Weighing a second part of chlorogenic acid and the chlorogenic acid phospholipid complex obtained in the step (1), mixing, and uniformly stirring to obtain the chlorogenic acid-containing pharmaceutical composition.
The mass ratio of the soybean lecithin, the first chlorogenic acid and 75% ethanol aqueous solution is 5:5:1000.
the mass ratio of the second part of chlorogenic acid to the chlorogenic acid phospholipid complex obtained in the step (1) is 1:2.
in each example and each figure, chlorogenic acid is abbreviated as CGA, chlorogenic acid phospholipid complex is abbreviated as CAPC, and a pharmaceutical composition containing chlorogenic acid is abbreviated as CA-CAPC.
Example 2
The preparation method of the chlorogenic acid-containing pharmaceutical composition comprises the following steps:
(1) Weighing dimyristoyl phosphatidylethanolamine and a first part of chlorogenic acid, placing into a container, adding 90 vol% ethanol water solution, stirring at 35 deg.C for 1h, recovering ethanol under reduced pressure (50 deg.C and-0.08 Mpa), drying the solid under reduced pressure, pulverizing the dried solid, and sieving with a No. six sieve to obtain chlorogenic acid phospholipid complex;
(2) Weighing a second part of chlorogenic acid and the chlorogenic acid phospholipid complex obtained in the step (1), mixing, and uniformly stirring to obtain the chlorogenic acid-containing pharmaceutical composition.
The mass ratio of dimyristoyl phosphatidyl ethanolamine to the first chlorogenic acid to the 90% ethanol aqueous solution is 10:2:4000.
the mass ratio of the second part of chlorogenic acid to the chlorogenic acid phospholipid complex obtained in the step (1) is 1:1.
example 3
The preparation method of the chlorogenic acid-containing pharmaceutical composition comprises the following steps:
(1) Weighing dimyristoyl phosphatidyl glycerol and a first part of chlorogenic acid, placing in a container, adding 80 vol.% ethanol water solution, stirring at 20 deg.C for 2h, recovering ethanol under reduced pressure (50 deg.C and-0.08 Mpa), drying the solid under reduced pressure, pulverizing the dried solid, and sieving with a No. four sieve to obtain chlorogenic acid phospholipid complex;
(2) Weighing a second part of chlorogenic acid and the chlorogenic acid phospholipid complex obtained in the step (1), mixing by an equivalent incremental method, and uniformly stirring to obtain the pharmaceutical composition containing chlorogenic acid.
The mass ratio of dimyristoyl phosphatidyl glycerol to the first chlorogenic acid to 80% ethanol aqueous solution is 4:10:2000.
the mass ratio of the second part of chlorogenic acid to the chlorogenic acid phospholipid complex obtained in the step (1) is 1:20.
example 4
The preparation method of the chlorogenic acid-containing pharmaceutical composition comprises the following steps:
(1) Weighing phospholipid (phospholipid is composed of distearoyl phosphatidic acid and dipalmitoyl phosphatidic acid with the mass ratio of 1:1) and a first part of chlorogenic acid, placing the mixture in a container, adding 70% ethanol aqueous solution, stirring at 30 ℃ for 2h, recovering ethanol under reduced pressure (25 ℃ and-0.08 Mpa), drying the solid under reduced pressure, crushing the dried solid, and sieving with a No. seven sieve to obtain a chlorogenic acid-phospholipid complex;
(2) Weighing a second part of chlorogenic acid and the chlorogenic acid phospholipid complex obtained in the step (1), mixing by an equivalent incremental method, and uniformly stirring to obtain the pharmaceutical composition containing chlorogenic acid.
The mass ratio of the phospholipid to the first chlorogenic acid to the 70% ethanol aqueous solution is 3:1:100.
the mass ratio of the second part of chlorogenic acid to the chlorogenic acid phospholipid complex obtained in the step (1) is 10:1.
example 5
The preparation method of the chlorogenic acid-containing pharmaceutical composition comprises the following steps:
(1) Weighing yolk lecithin and a first part of chlorogenic acid, placing into a container, adding 75 vol% ethanol water solution, stirring at 30 deg.C for 2h, recovering ethanol under reduced pressure (25 deg.C and-0.08 Mpa), drying the solid under reduced pressure, pulverizing the dried solid, and sieving with a third sieve to obtain chlorogenic acid phospholipid complex;
(2) Weighing a second part of chlorogenic acid and the chlorogenic acid phospholipid complex obtained in the step (1), mixing, and uniformly stirring to obtain the chlorogenic acid-containing pharmaceutical composition.
The mass ratio of the egg yolk lecithin, the first chlorogenic acid and the ethanol water solution with the volume concentration of 70% is 1:7:500.
the mass ratio of the second part of chlorogenic acid to the chlorogenic acid phospholipid complex obtained in the step (1) is 10:20.
in place of the soybean phospholipid of example 1, dimyristoyl lecithin, dipalmitoyl lecithin, distearoyl lecithin, dioleoyl lecithin, 1-myristoyl-2-palmitoyl lecithin, 1-stearoyl-2-myristoyl lecithin, myristoyl lysolecithin, palmitoyl lysolecithin, stearoyl lysolecithin, dilauroyl phosphatidylglycerol, 1,2-palmitoyl phosphatidylglycerol, distearoyl phosphatidylglycerol, dimyristoyl phosphatidic acid, dioleoyl phosphatidylethanolamine, dipalmitoyl phosphatidylethanolamine, distearoyl phosphatidylethanolamine, 1-palmitoyl-2-oleoyl phosphatidylethanolamine, dimyristoyl phosphatidylserine, dioleoyl phosphatidylserine, dipalmitoyl phosphatidylserine, or distearoyl phosphatidylserine, and otherwise as in example 1, a chlorogenic acid-containing pharmaceutical composition having properties and effects similar to those of example 1 was prepared, respectively.
To further verify the excellent effects of the present invention, the inventors also performed two types of experiments on the chlorogenic acid-containing pharmaceutical composition of the present invention (prepared in example 1): the first type is oral pharmacokinetic experiments carried out on rats, which proves that the pharmaceutical composition containing chlorogenic acid of the invention improves the oral bioavailability and the pharmacokinetic parameters; the second type is a synergistic pharmacodynamic experiment, a lipopolysaccharide induced mouse acute lung injury model is adopted, and the influence of the medicament on pharmacodynamic indexes such as mouse survival rate, weight change, lung tissue inflammatory factor expression, respiratory function and the like is inspected; the advantages and excellent effects of the chlorogenic acid-containing pharmaceutical composition in the treatment and/or prevention of acute lung injury are proved from two aspects of oral bioavailability and pharmacodynamics.
Through the coordination of the preparation process, the preparation concentration of the test sample liquid medicine and the administration dose, the absolute administration doses of chlorogenic acid of three groups of rats are kept consistent in rat oral pharmacokinetic research test, the absolute administration doses of chlorogenic acid in mouse acute lung injury pharmacodynamic test are kept consistent, the bioavailability and the pharmacodynamics of raw materials of the chlorogenic acid, a chlorogenic acid phospholipid compound and the pharmaceutical composition containing the chlorogenic acid are inspected under the condition of ensuring the consistency of the absolute administration doses of the chlorogenic acid, and the method and the results are as follows:
experimental example 1
Oral pharmacokinetic experiments in rats on chlorogenic acid (CGA), chlorogenic Acid Phospholipid Complex (CAPC), pharmaceutical composition of chlorogenic acid of the present invention (CA-CAPC)
The Chlorogenic Acid Phospholipid Complex (CAPC) used in this experiment was prepared by the step (1) of example 1; a pharmaceutical composition containing chlorogenic acid (CA-CAPC) was prepared as described in example 1. The chlorogenic acid phospholipid complexes in the experiment are all abbreviated as CAPC, and the pharmaceutical compositions containing chlorogenic acid are all abbreviated as CA-CAPC.
An experimental instrument:
an Agilent 6460Triple Quad LC/MS Triple quadrupole tandem mass spectrometer; a constant-temperature magnetic heating stirrer (instrument factory in south of the Yangtze river, gold jar city); HH-9 digital display constant temperature water bath (Suzhou Jiangdong precision instruments Co., ltd.); centrifuge (Beijing boao classical biotechnology limited); KQ-700B ultrasonic cleaner (Shanghai Hank scientific instruments, inc.); milli-Q ultra pure water instruments (MILLIPORE, USA; MS198 DU); electronic balance (Zhejiang Lichen instruments science and technology Co., ltd.); an SHZ-CB circulating water type multipurpose vacuum pump (Shanghai Yue Instrument Co., ltd.); one in ten thousand electronic balance (mettler-toledo instruments ltd, switzerland); one in ten million electronic balances (mertler-toledo instruments ltd, switzerland);
experimental reagent:
chlorogenic acid standard, puerarin standard (Zhengdong science and technology, purity 98%), soybean phospholipid (Shanghai Ai Weita medicine science and technology, inc.), chromatographic grade acetonitrile, methanol, formic acid (Fisher, USA), absolute ethanol, and ethanol with series concentration as analytically pure (Shijiazhuang, fourth pharmaceutical factory). The phospholipids referred to in the present invention are all commercial products.
Experimental animals:
SD rats, male, body weight (200. + -.20 g), purchased from Stevens Bei Fu (Beijing) Biotechnology Ltd. License number: SCXK (Jing) 2019-0010.
Chlorogenic acid analysis method chromatographic conditions:
an Aichcrombood AQ C18 column (250 mm. Times.4.6 mm. Times.5 μm) was used, and acetonitrile (B) -0.1% formic acid water (A) was used as a mobile phase, and gradient elution was carried out by using a mobile phase gradient, the procedure of which is shown in Table 1. The detection wavelength is 330nm, the column temperature is 30 ℃, the flow rate is 0.2mL/min, and the sample injection amount is 5 mu L.
TABLE 1 mobility gradient elution procedure
Chlorogenic acid analysis method mass spectrum parameters:
ESI electrospray ionization source with puerarin as internal standard; a positive ion detection mode; multiple reaction detection (MRM); ion source ejection voltage: -4000V; temperature of atomizing gas: 300 ℃; temperature of drying gas: 500 ℃; temperature of sheath gas: 400 ℃; collision gas, sheath gas, auxiliary gas: nitrogen gas; atomizer, heater, dryer pressure: 40. 50, 10psi; other parameters are shown in table 2.
TABLE 2 Mass Spectrometry parameter settings for chlorogenic acid and internal Standard Puerarin
Preparation of chlorogenic acid phospholipid complex:
prepared from example 1, step (1).
The preparation of the chlorogenic acid-containing pharmaceutical composition of the invention comprises the following steps:
prepared from example 1.
The preparation method of the test solution of chlorogenic acid raw material medicine comprises the following steps:
weighing appropriate amount of chlorogenic acid raw material medicine, adding 30 vol% ethanol water solution, and making into 30mg/ml medicinal liquid.
The preparation method of the test sample liquid medicine of the chlorogenic acid phospholipid complex comprises the following steps:
weighing appropriate amount of chlorogenic acid phospholipid complex, adding 30% ethanol water solution, and making into 60mg/ml medicinal liquid.
The preparation method of the test solution of the chlorogenic acid-containing pharmaceutical composition comprises the following steps:
weighing a proper amount of the pharmaceutical composition containing chlorogenic acid, and adding 30% ethanol aqueous solution by volume concentration to prepare a liquid medicine with the concentration of 45 mg/ml.
Oral pharmacokinetic study in rats:
18 SD male mice were randomly divided into 3 groups of 6, each group consisting of chlorogenic acid group (CGA, administration dose of 150 mg/kg), chlorogenic acid-phospholipid complex group (CAPC, administration dose of 300 mg/kg) and chlorogenic acid-containing pharmaceutical composition group (CA-CAPC, administration dose of 225 mg/kg).
Feeding normally for one week, fasting for 12h before experiment, only drinking water, and separately intragastrically administering chlorogenic acid raw material drug, chlorogenic acid phospholipid complex and chlorogenic acid-containing pharmaceutical composition (i.e. intragastrically administering 1mL reagent per 200g rat body weight) according to dosage of 1mL/200 g.
Respectively taking blood from eye sockets after administration for 5min,10min,15min,20min,30min,45min,1h,1.5h,2h,3h,4h,6h and 8h, then quickly filling the blood into a heparinized 1.5mL centrifuge tube, shaking, centrifuging for 10min at 10000r/min, separating plasma, and storing in a refrigerator at-80 ℃ for later use.
Taking 100 mu L of upper plasma in a 1.5mL centrifuge tube, adding 10 mu L of methanol solution of internal standard (containing 500ng/mL puerarin) and 280 mu L of acetonitrile, performing vortex oscillation for 2min, centrifuging at 10000r/min for 10min, and taking 5 mu L of supernatant for sample injection. And (4) sampling and measuring according to the chromatographic condition and mass spectrum condition of the chlorogenic acid analytical method, and drawing an average blood concentration-time curve of the chlorogenic acid according to the measurement result. The linearity, precision, repeatability, stability, recovery rate and matrix effect of the method used in content determination are verified, and the content determination requirement is met.
Results of the experiment
Mean plasma concentration-time curves of different groups of chlorogenic acids are shown in figure 1.
The major pharmacokinetic parameters were analyzed and calculated using WinNolin version 6.2 software (Certara, USA), and the major parameters of plasma pharmacokinetics were calculated for three groups of animals according to a non-compartmental model parameter estimation method, and the results are shown in Table 3.
TABLE 3 major pharmacokinetic parameters of chlorogenic acid after oral administration of each group of drugs
Note: compared with chlorogenic acid bulk drug, p is less than 0.05, p is less than 0.01, p is less than 0.001, and the difference has statistical significance.
Compared with the chlorogenic acid phospholipid complex, the preparation method has the advantages that, # p＜0.05， ## p＜0.01， ### p is less than 0.001, and the difference has statistical significance.
As can be seen from fig. 1 and table 3, the chlorogenic acid crude drug has low drug concentration in the absorption phase and low oral bioavailability after oral administration.
As can be seen from FIG. 1 and Table 3, compared with the chlorogenic acid bulk drug, AUC (0-t) and AUC (0- ∞) of the chlorogenic acid phospholipid complex are both significantly improved (p < 0.001, p < 0.01), which indicates that the oral bioavailability of the chlorogenic acid is significantly improved after the chlorogenic acid is prepared into the chlorogenic acid phospholipid complex. However, the pharmacokinetic parameters of chlorogenic acid phospholipid complexes have significant disadvantages: firstly, the Tmax of the chlorogenic acid phospholipid complex is obviously increased, the peak reaching time is delayed, and the onset of the chlorogenic acid is possibly slowed; secondly, the concentration of the drug in the absorption phase of the chlorogenic acid phospholipid complex is lower than that of the chlorogenic acid raw material drug, so that the aim of quick response cannot be fulfilled.
As can be seen from fig. 1 and table 3, compared with the chlorogenic acid bulk drug, both AUC (0-t) and AUC (0- ∞) of the pharmaceutical composition containing chlorogenic acid are significantly improved (improved by 41.9%, 59.5%) (p < 0.05 ), which indicates that the oral bioavailability of chlorogenic acid can be significantly improved after the chlorogenic acid is prepared into the pharmaceutical composition containing chlorogenic acid; meanwhile, the medicine concentration of the absorption phase of the medicine composition containing the chlorogenic acid is obviously higher than that of a chlorogenic acid raw material (p is less than 0.05), the peak reaching time is obviously faster than that of the chlorogenic acid raw material medicine (p is less than 0.05), and the maximum blood medicine concentration Cmax is equal to the chlorogenic acid raw material medicine and has no obvious difference. Therefore, the pharmaceutical composition containing the chlorogenic acid improves the oral bioavailability of the chlorogenic acid, improves the pharmacokinetic parameters of the chlorogenic acid, ensures that the blood concentration of the absorption phase of the chlorogenic acid after administration quickly reaches a higher concentration and quickly reaches the peak, and is beneficial to the treatment of diseases.
As can be seen from FIG. 1 and Table 3, the AUC (0-t) and AUC (0- ∞) of the pharmaceutical composition containing chlorogenic acid have no significant difference from the chlorogenic acid phospholipid complex, the drug concentration of the absorption phase of the pharmaceutical composition containing chlorogenic acid is significantly higher than that of the chlorogenic acid phospholipid complex (p < 0.01), the time to peak is significantly faster than that of the chlorogenic acid phospholipid complex (p < 0.001), and the maximum blood drug concentration Cmax is significantly higher than that of the chlorogenic acid phospholipid complex (p < 0.05). Therefore, the pharmaceutical composition containing the chlorogenic acid not only improves the oral bioavailability of the chlorogenic acid, but also enables the blood concentration to quickly reach the peak, has high absorption phase concentration, greatly improves the defect of the pharmacokinetic parameters of the chlorogenic acid phospholipid complex, and provides a powerful pharmacokinetic theoretical basis for treating acute diseases by the chlorogenic acid.
In view of the above experimental results, the pharmaceutical composition containing chlorogenic acid of the present invention achieves a significant advantage in terms of pharmacokinetics: the oral bioavailability is improved, the absorption peak is fast, the absorption phase drug concentration is high, the drug peak value concentration is high, the pharmacokinetic parameters of the chlorogenic acid and the chlorogenic acid phospholipid compound are integrally improved, and the theoretical basis of pharmacokinetics and the pharmaceutical basis for treating acute lung injury by the chlorogenic acid are provided.
Experimental example 2
Research on pharmacodynamics test of protective effect of chlorogenic acid (CGA), chlorogenic Acid Phospholipid Complex (CAPC) and chlorogenic acid-containing pharmaceutical composition (CA-CAPC) on acute lung injury mice
The chlorogenic acid-phospholipid complex (CAPC) used in the experiment was prepared in the step (1) of example 1, and the mass ratio of chlorogenic acid to phospholipid was 1:1; a pharmaceutical composition containing chlorogenic acid (CA-CAPC) was prepared as in example 1, and the mass ratio of chlorogenic acid to chlorogenic acid phospholipid complex was 1:2. The chlorogenic acid phospholipid complexes in the experiment are all abbreviated as CAPC, and the pharmaceutical compositions containing chlorogenic acid are all abbreviated as CA-CAPC.
The experimental method comprises the following steps:
6-8 week male C57BL/6N mice were randomly divided into 6 groups of 16 mice each, which were blank (Control, abbreviated con), model (Model, abbreviated M or LPS,5 mg/kg), positive drug dexamethasone Control (Positive, abbreviated pos or LPS + DEX,5mg/kg, sigma), chlorogenic acid (CGA, or LPS + CGA,100 mg/kg), chlorogenic acid phospholipid complex (CAPC, or LPS + CAPC,200 mg/kg), chlorogenic acid-containing pharmaceutical composition of the present invention (CA-CAPC, or LPS + CA-CAPC,150 mg/kg).
Chlorogenic acid (CGA) was purchased from Shanghai Allantin Biotechnology, inc.
And (4) investigation indexes are as follows:
mouse survival rate, mouse weight, mouse respiratory function evaluation and mouse lung tissue inflammatory factor expression.
The reagent preparation method comprises the following steps:
preparation of lipopolysaccharide (LPS, sigma Co.) solution: 10mg of LPS solid was precisely weighed and sufficiently dissolved in 10mL of ultrapure water to obtain a sample of LPS solution at a concentration of 1mg/mL.
Positive drug dexamethasone control DEX (0.5 mg/mL): 5mg of dexamethasone (DEX, sigma) was weighed out and dissolved well in 10mL of ultrapure water at a concentration of 0.5mg/mL.
Chlorogenic acid group CGA (10 mg/mL) is prepared by weighing chlorogenic acid 100mg, and dissolving in 10mL ultrapure water to obtain chlorogenic acid liquid medicine with concentration of 10mg/mL.
CAPC (20 mg/mL) is prepared by weighing CAPC 200mg, and dissolving in 10mL ultrapure water to obtain CAPC medicinal liquid with concentration of 20mg/mL.
The chlorogenic acid-containing pharmaceutical composition group CA-CAPC (15 mg/mL) of the invention is prepared by weighing 150mg of CA-CAPC and fully dissolving the CA-CAPC in 10mL of ultrapure water to obtain a CA-CAPC liquid medicine with the concentration of 15mg/mL.
Modeling, administration mode and dosage, and experimental operation:
the mice in the blank group and the model group are respectively perfused with 0.9 percent physiological saline solution (Shijiazhu fourth pharmaceutical factory) 0.1mL/10g (namely perfusing 0.1mL reagent per 10g of the weight of the mice, the same is carried out below), the mice in the positive drug control group (pos) are perfused with 0.1mL/10g of dexamethasone solution (DEX), the mice in the administration group are separately perfused with 0.1mL/10g of chlorogenic acid solution (CGA), 0.1mL/10g of chlorogenic acid phospholipid complex group (CAPC) and 0.1mL/10g of the pharmaceutical composition group (CA-CAPC) containing the chlorogenic acid. The test mice were weighed daily and the weight changes of the mice were examined. All groups of mice were gavaged for 2 days.
On day 3, LPS solution was injected into the tail vein of mice in each group except the blank group at a rate of 0.1mL/20g for 3 consecutive days to model the mice.
On the 6 th day, the LPS solution injection is stopped, and the mice of each group are continuously administered with the 0.9 percent physiological saline solution, the dexamethasone, the chlorogenic acid phospholipid complex and the pharmaceutical composition containing the chlorogenic acid for 2 days by intragastric administration.
Counting the survival rate of the mice on day 7; detecting the respiratory function of the experimental mouse; then, lung tissue is taken and lung tissue inflammatory factor expression is measured.
And (3) detecting the respiratory function of the mouse: before the material is taken on the 7 th day, an animal lung function monitoring system (EMKA-WBP, france EMKA technoLogies company) is used for detecting the respiratory function of the experimental mice, and the detection method is the lung function monitoring-whole plethysmography (WBP) of the animal in the waking state without constraint. The animals are placed in a trace box, the detection is started after the animals are stable and inactive (about 10 minutes), the data is recorded every ten seconds, and the data from the 5 th minute to the 10 th minute after the detection is selected for statistics. During detection, the room is quiet, the temperature and the humidity are kept constant, the airflow change is small, and the interference to animals is reduced as much as possible. The detection indexes are as follows: tidal Volume (TV), ventilation per Minute (MV). The influence of chlorogenic acid (CGA), chlorogenic acid-phospholipid complex (CAPC) and the pharmaceutical composition containing the chlorogenic acid (CA-CAPC) on the respiratory function of the mice is researched by a pretreatment group/delta Model group method (Treatment/delta Model method), and the Treatment/delta Model method comprises the following specific steps: (1) The lung function Blank data (initial lung function data before intervention of each group) of each administration group is subjected to model homogenization treatment, and the treatment method comprises the following steps: blank normalized value = mean of Blank values for each dosing group/mean of Blank values for model group; (2) The Treatment method comprises the following steps of calibrating the Treatment group lung function data of each administration group with a Blank group: treatment calibration value = Treatment group value/Blank normalized value for each administration group; (3) Finally, the calibrated data is subjected to Model set homogenization Treatment, and the result is expressed by an intervention set/delta Model set method (maintenance/delta Model), namely maintenance/delta Model = maintenance calibration value/maintenance Model set average.
Enzyme-linked immunosorbent assay (ELISA) for detecting inflammatory factors of lung tissues: shearing 20mg of mouse lung tissue, placing the mouse lung tissue into a 1.5mL centrifuge tube, adding 180 mu L of normal saline, and ultrasonically crushing the tissue. After the ELISA kit (R & D Systems, USA) was left to stand at room temperature for 30min, the detection of inflammatory factors IL-6, TNF-alpha and IL-1 beta was carried out by a multifunctional microplate reader (Spark, TECAN, switzerland) according to the instructions.
The research results are as follows:
the survival rate of the mice is shown in fig. 2 after the mice are subjected to LPS-induced acute lung injury model and drug-induced prognosis. After the drug dry prognosis, the survival rate of the blank group of mice is unchanged;
the survival rate of the model group (LPS) on day 6 (day 3 after LPS treatment) was reduced to 50%;
the survival rate on day 6 (day 3 after LPS treatment) of the positive dexamethasone control group administered (pos) was 100%;
chlorogenic acid group (CGA, or LPS + CGA), survival rate of 94% on 6 days of administration (3 days after LPS treatment);
the survival rates of mice after administration of the chlorogenic acid phospholipid complex group (CAPC, or LPS + CAPC) and the chlorogenic acid-containing pharmaceutical composition group (CA-CAPC, or LPS + CA-CAPC) of the present invention were consistent with those of the blank group, and both were 100%.
The results indicate that LPS can cause death in mice; chlorogenic acid (CGA), a Chlorogenic Acid Phospholipid Complex (CAPC) and the pharmaceutical composition containing the chlorogenic acid (CA-CAPC) can delay the death time of mice caused by LPS and improve the survival rate of the mice, wherein the survival rate of the mice improved by the Chlorogenic Acid Phospholipid Complex (CAPC) and the pharmaceutical composition containing the chlorogenic acid (CA-CAPC) is higher than that of the chlorogenic acid (CGA) (see figure 2). The chlorogenic acid-containing pharmaceutical composition (CA-CAPC) provided by the invention has a remarkable effect of improving the survival rate of mice caused by LPS.
After LPS and drug-dried prognosis, the weight of each group of mice is shown in FIG. 3-A, and the weight loss is shown in FIG. 3-B. After the mice are dried by different medicines, the weight average of the bodies of the mice in the first 3 days is not obviously changed; after LPS intervention, the body weight of mice in each group was significantly reduced (p < 0.05) except for the blank group (which was given an equal amount of physiological saline). After a positive drug dexamethasone (pos), a chlorogenic acid (CGA), a Chlorogenic Acid Phospholipid Complex (CAPC) and a pharmaceutical composition containing the chlorogenic acid (CA-CAPC) are given, compared with a model group, the weight loss of mice of the positive drug dexamethasone control group (pos), the chlorogenic acid group (CGA), the chlorogenic acid phospholipid complex group (CAPC) and the pharmaceutical composition containing the chlorogenic acid (CA-CAPC) is remarkably relieved, as shown in figure 3-A, the weight loss is remarkably reduced (p is less than 0.05, p is less than 0.01, p is less than 0.001), as shown in figure 3-B; compared with the chlorogenic acid group (CGA), the weight loss of the mice of the chlorogenic acid-containing pharmaceutical composition group (CA-CAPC) is remarkably relieved, as shown in figure 3-A, the weight loss is remarkably reduced (p is less than 0.001), as shown in figure 3-B; compared with the chlorogenic acid phospholipid complex group (CAPC), the weight loss of the mice of the chlorogenic acid-containing pharmaceutical composition group (CA-CAPC) is remarkably relieved, as shown in figure 3-A, the weight loss is remarkably reduced (p is less than 0.05), as shown in figure 3-B. The pharmaceutical composition containing chlorogenic acid (CA-CAPC) of the invention has the most obvious effect of protecting the weight of mice, and is remarkably stronger than chlorogenic acid (CGA) and Chlorogenic Acid Phospholipid Complex (CAPC) (p is less than 0.05, and p is less than 0.001) (see figure 3-A and figure 3-B).
After the mice are subjected to LPS and drug drying prognosis, the lung tissue inflammatory factor results of each group of mice are shown in figure 4-A, figure 4-B and figure 4-C, and after LPS is given, the expressions of the lung tissue inflammatory factors IL-6, TNF-alpha and IL-1 beta of the mice are obviously increased; after positive drugs of dexamethasone (pos), chlorogenic acid (CGA), chlorogenic Acid Phospholipid Complex (CAPC) and the pharmaceutical composition containing chlorogenic acid (CA-CAPC) are given, the expression of each inflammatory factor of lung tissues of mice is obviously reduced. Compared with the chlorogenic acid group (CGA), the mouse lung tissue inflammatory factor IL-6 expression of the chlorogenic acid-containing pharmaceutical composition group (CA-CAPC) is remarkably reduced (p is less than 0.05), as shown in FIG. 4-A, the mouse lung tissue inflammatory factor TNF-alpha expression is remarkably reduced (p is less than 0.05), as shown in FIG. 4-B, the mouse lung tissue inflammatory factor IL-1 beta expression is remarkably reduced (p is less than 0.05), as shown in FIG. 4-C; compared with the chlorogenic acid phospholipid complex group (CAPC), the mouse lung tissue inflammatory factor IL-6 expression of the chlorogenic acid-containing pharmaceutical composition group (CA-CAPC) is remarkably reduced (p is less than 0.05), as shown in figure 4-A, and the mouse lung tissue inflammatory factor IL-1 beta expression is remarkably reduced (p is less than 0.05), as shown in figure 4-C. The chlorogenic acid-containing pharmaceutical composition (CA-CAPC) has stronger effects of reducing lung inflammatory factors and resisting inflammatory storm, and the effects are stronger than those of chlorogenic acid (CGA) and Chlorogenic Acid Phospholipid Complex (CAPC). The chlorogenic acid-containing pharmaceutical composition (CA-CAPC) can obviously reduce lung inflammatory factors and has the effects of preventing and treating inflammatory storm induced by LPS.
Respiratory function is an important macroscopic indicator of lung function and is a golden indicator for evaluating drug protection lung injury. The research adopts the respiratory function index to evaluate the treatment effect of the medicament on the acute lung injury mice, and is important evidence for treating the acute lung injury by chlorogenic acid and a composition containing the chlorogenic acid. After LPS and drug drying, the tidal volume-time curve, tidal volume, minute ventilation-time curve and minute ventilation of each group of mice are shown in figure 5-A, figure 5-B, figure 5-C and figure 5-D, and after LPS molding, the tidal volume and minute ventilation of each group of mice are weakened. Compared with the model group, the positive drug dexamethasone control group (pos), the chlorogenic acid group (CGA), the chlorogenic acid phospholipid complex group (CAPC) and the chlorogenic acid-containing pharmaceutical composition group (CA-CAPC) administration group of the invention can significantly up-regulate the tidal volume (p is less than 0.01, p is less than 0.001), and significantly up-regulate the ventilation per minute (p is less than 0.01, p is less than 0.05, p is less than 0.01, p is less than 0.001). Compared with the group of chlorogenic acids (CGA), the mouse tidal volume of the group of chlorogenic acid-containing pharmaceutical composition (CA-CAPC) of the present invention was significantly increased (p < 0.05), and the ventilation per minute was significantly increased (p < 0.05), as shown in FIG. 5-B and FIG. 5-D. With the chlorogenic acid phospholipid complex group (CAPC), the mouse tidal volume of the chlorogenic acid-containing pharmaceutical composition group (CA-CAPC) of the present invention was significantly increased (p < 0.05), and the ventilation per minute was significantly increased (p < 0.05), as shown in FIG. 5-B and FIG. 5-D. The chlorogenic acid group (CGA), the chlorogenic acid phospholipid complex group (CAPC) and the chlorogenic acid-containing pharmaceutical composition group (CA-CAPC) can inhibit lung injury respiratory failure caused by LPS and restore the lung respiratory function, wherein the prevention and treatment effects of the chlorogenic acid-containing pharmaceutical composition group (CA-CAPC) are stronger than those of the chlorogenic acid group (CGA) and the chlorogenic acid phospholipid complex group (CAPC).
The results of comparison of the relative values of the tidal volume change and the relative value of the ventilation per minute of the intervention group/delta Model group (Treatment/delta Model) of the chlorogenic acid-containing composition (CA-CAPC) group are shown in FIG. 6-A and FIG. 6-B, and analysis shows that compared with the chlorogenic acid (CGA), the chlorogenic acid-containing composition (CA-CAPC) can significantly improve the relative value of the tidal volume change (p is less than 0.05) of an LPS lung injury mouse and significantly improve the relative value of the ventilation per minute (p is less than 0.001) of the LPS lung injury mouse; compared with Chlorogenic Acid Phospholipid Complex (CAPC), the chlorogenic acid-containing composition (CA-CAPC) can remarkably improve the relative tidal volume change value (p is less than 0.05) of LPS lung injury mice and remarkably improve the relative ventilation change value per minute (p is less than 0.05) of the LPS lung injury mice; the chlorogenic acid-containing composition (CA-CAPC) has good effects of resisting lung respiratory failure and recovering lung respiratory function, and the effects are stronger than those of chlorogenic acid (CGA) and Chlorogenic Acid Phospholipid Complex (CAPC).
Experiments prove that the pharmacokinetic and pharmacodynamic experimental results of the pharmaceutical compositions containing chlorogenic acid prepared in examples 2, 3, 4 and 5 are similar to the experimental results of the pharmaceutical composition containing chlorogenic acid prepared in example 1.
Claims (5)
1. The preparation method of the chlorogenic acid-containing pharmaceutical composition is characterized by comprising the following steps:
(1) Weighing phospholipid and a first part of chlorogenic acid, placing in a container, adding 75% ethanol water solution, stirring at 20-35 deg.C for 1-2h, recovering ethanol under reduced pressure, drying the solid under reduced pressure, pulverizing, and sieving to obtain chlorogenic acid-phospholipid complex;
(2) Weighing a second part of chlorogenic acid and the chlorogenic acid phospholipid complex obtained in the step (1), mixing or mixing according to an equivalent incremental method, and uniformly stirring to obtain a pharmaceutical composition containing chlorogenic acid;
the mass ratio of the phospholipid to the first chlorogenic acid to 75% ethanol aqueous solution is 5:5:1000, parts by weight;
the mass ratio of the second part of chlorogenic acid to the chlorogenic acid phospholipid complex obtained in the step (1) is 1:2;
the phospholipid is soybean phospholipid.
2. The method of claim 1, wherein the screening is through a third screen to a seventh screen.
3. The method of claim 2, wherein the screen is a size five screen.
4. A pharmaceutical composition comprising chlorogenic acid prepared by the method of any of claims 1-3.
5. Use of the chlorogenic acid-containing pharmaceutical composition of claim 4 in preparing a medicament for preventing and treating acute lung injury.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110265517.3A CN113041352B (en) | 2021-03-11 | 2021-03-11 | Chlorogenic acid-containing pharmaceutical composition, preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110265517.3A CN113041352B (en) | 2021-03-11 | 2021-03-11 | Chlorogenic acid-containing pharmaceutical composition, preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113041352A CN113041352A (en) | 2021-06-29 |
| CN113041352B true CN113041352B (en) | 2023-01-17 |
Family
ID=76511401
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110265517.3A Active CN113041352B (en) | 2021-03-11 | 2021-03-11 | Chlorogenic acid-containing pharmaceutical composition, preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113041352B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020253689A1 (en) * | 2019-06-17 | 2020-12-24 | 四川九章生物科技有限公司 | Chlorogenic acid self-emulsifying composition and use thereof |
-
2021
- 2021-03-11 CN CN202110265517.3A patent/CN113041352B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020253689A1 (en) * | 2019-06-17 | 2020-12-24 | 四川九章生物科技有限公司 | Chlorogenic acid self-emulsifying composition and use thereof |
Non-Patent Citations (2)
| Title |
|---|
| 赵安权等.绿原酸磷脂复合物制备工艺的研究.《华西药学杂志》.2015,(第02期), * |
| 郭静等.绿原酸对小鼠急性肺损伤的保护作用研究.《天然产物研究与开发》.2018,(第07期),第1214-1218页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113041352A (en) | 2021-06-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2010500342A (en) | Polar organic extract of Eurocomalongifolia | |
| CN108938654B (en) | Pulsatillae saponin B4 injection preparation | |
| AU2017422167B2 (en) | Preparation of Pulsatilla saponin B4 for injection | |
| CN109846896B (en) | Application of hederagenin in preparation of medicine for resisting inflammatory injury of vascular endothelial cells | |
| Xu et al. | Pharmacokinetics of eight flavonoids in rats assayed by UPLC-MS/MS after oral administration of Drynariae rhizoma extract | |
| WO2023024804A1 (en) | Compound dry powder inhaler and application thereof | |
| CN113041352B (en) | Chlorogenic acid-containing pharmaceutical composition, preparation method and application thereof | |
| CN113143903B (en) | Application of chlorogenic acid-containing pharmaceutical composition in preparation of antiviral drugs | |
| Guo et al. | Pharmacokinetic comparisons of mangiferin and mangiferin monosodium salt in rat plasma by UPLC-MS/MS | |
| Chen et al. | Xiao-chai-hu decoction ameliorates poly (I: C)-Induced viral pneumonia through inhibiting inflammatory response and modulating serum metabolism | |
| CN113209111A (en) | Application of brown algae oligosaccharide | |
| WO2019134159A1 (en) | Rectal mucosal administration preparation of pulsatilla chinensis (bge.) regel saponin b4 and preparation method therefor | |
| CN113116907A (en) | Medical application of cycloastragenol | |
| KR20180073557A (en) | Composition and method thereof | |
| JPH0137377B2 (en) | ||
| EP4265260A1 (en) | Stable liquid pharmaceutical composition containing kuding saponin compound | |
| Zhao et al. | Pharmacokinetic study of coadministration with cefuroxime sodium for injection influencing ReDuNing injection-derived seven phytochemicals and nine metabolites in rats | |
| Yu et al. | Study of the therapeutic effects of Painong powder on ulcerative colitis and the role of Platycodonis Radix in the prescription based on pharmacodynamic, pharmacokinetic, and tissue distribution analyses | |
| US10406191B2 (en) | Compounds, extracts and methods from chinese medicinal herb sophora flavescens that inhibit airway contraction | |
| EP1498131B1 (en) | Medicinal preparation containing phenylethanoid glycosides extracted from Cistanche tubulosa | |
| CN112028867B (en) | Tricyclic diterpene compound and extraction method and application thereof | |
| EP4344704A1 (en) | Ganoderma spore oil and use thereof in preparation of drug counteracting cancer-related fatigue | |
| CN115429812B (en) | Monnpiravir phospholipid complex and preparation method thereof | |
| CN117462583A (en) | Preparation method of radix codonopsis pilosulae oral liquid for treating acute lung injury | |
| CN112824428B (en) | Biological peptide for treating lung diseases and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |