CN111100177B - (20S,24R) -ocotillol type ginsenoside glycine derivative, preparation method and application thereof - Google Patents
(20S,24R) -ocotillol type ginsenoside glycine derivative, preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a new compound (20S,24R) -ocotillol type ginsenoside glycine derivative, a preparation method and application thereof, belonging to the field of medicines. The chemical name of the compound is (20S,24R) -3-O-aminoacetyl-dammar-20, 24-epoxy-3 beta, 6 alpha, 12 beta, 25-tetrol. Taking an appropriate amount of Ocotillol, reacting with BOC-Gly, DMAP, EDCI and DIPEA at room temperature, diluting with ethyl acetate, extracting with ethyl acetate, combining organic phases, dissolving in dichloromethane after column chromatography purification, adding TFA, reacting at room temperature, extracting with dichloromethane, combining the organic phases, and purifying by column chromatography to obtain the compound. The experimental result proves that the (20S,24R) -ocotillol type ginsenoside glycine derivative has protective effect on the lung function of experimental model mice of hormone-resistant asthma diseases caused by non-typeable haemophilus influenzae (NTHi) and can improve the hormone resistance of the experimental model mice. The (20S,24R) -ocotillol ginsenoside glycine derivative is used as a medicinal active ingredient, provides clinical guidance and basis for preventing and treating hormone-resistant asthma, improves the drug resistance of patients to hormone, further relieves the pain of the patients and improves the life quality of the patients.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to a (20S,24R) -ocotillol ginsenoside glycine derivative, an extraction method thereof and application thereof in preparing a medicine for treating hormone-resistant asthmatic diseases.
Background
Asthma is a chronic inflammatory disease of the airways characterized by a high airway response and reversible airflow limitation. The clinical manifestations are cough, expectoration, recurrent wheezing, chest tightness and dyspnea, with intensity varying with time, usually with triggers such as antigen or irritant exposure, seasonal changes or viral respiratory infections. Asthma is a common chronic respiratory disease with high incidence rate, more than 3 hundred million asthma patients are in the world at present, and the incidence rate is increased year by year. The disease cannot be cured radically, needs to be managed in a grading way according to the severity of asthma and insist on long-term treatment, and the current clinical medicine for treating asthma is to treat the asthma by taking glucocorticoid as a first line and matching beta receptor stimulant, anticholinergic medicine, theophylline medicine, leukotriene regulator, antihistamine medicine and the like. However, clinical treatment has a problem that the disease is a heterogeneous disease, which is divided into different subtypes, wherein hormone-resistant asthma is characterized by more lasting attack, more serious disease and poor response to first-line glucocorticoid treatment, and although the subtype accounts for 5-20% of the general population of asthma, the subtype consumes more than 50-80% of the asthma-related medical resources, and the hospitalization rate and the death rate of the subtype are significantly higher than those of patients with other asthma subtypes. Studies report that hormone-resistant asthma subtypes develop in association with bacterial infection, oxidative stress, and airway remodeling due to chronic inflammation of the airways.
American ginseng (Panax quinquefolium L.) is native to the eastern United states and Canada, is a herbaceous plant of Panax genus of Araliaceae family, also called American ginseng, American ginseng and Guangdong ginseng, and has been cultivated in China recently. American ginseng is bitter in taste and cool in nature, has the effects of tonifying lung yin, clearing fire, nourishing stomach and promoting fluid production, and is used for treating lung and stomach fluid deficiency, hectic fever, lung deficiency, hemoptysis, qi deficiency and the like. The main active components of the American ginseng are triterpenoid saponins, and the ginsenosides can be divided into the following three types according to the difference of sapogenin structures: dammarane-type tetracyclic triterpene saponin, oxtriptolone-type tetracyclic triterpene saponin and oleanolic acid-type pentacyclic triterpene saponin. Research reports indicate that the ocrtalone ginsenoside 12-one-PF11 can be reducedLight H2O2The resulting oxidative stress damage of cells; the oxtriptolone type ginsenoside ocotillol can improve the oxidative stress injury of mice and reduce the inflammation level; in addition, the ocotillol type ginsenoside also has strong antibacterial activity on gram-positive bacteria and can enhance the antibacterial effect of other antibiotics.
Glycine, also known as glycine, is an amino acid that is not essential to the human body, has both acidic and basic functional groups in the molecule, has strong hydrophilicity, and participates in the synthesis of proteins and physiologically active molecules associated with many important metabolisms. In recent years, researches show that glycine can improve oxidative stress injury of various animal models and has a wide anti-inflammatory effect; in addition, glycine can also reduce skin allergic reaction of a mouse model and has an antiallergic effect.
Disclosure of Invention
The invention aims to provide a (20S,24R) -ocotillol ginsenoside glycine derivative, a preparation method and a medical application thereof.
The technical scheme adopted by the invention is as follows:
(20S,24R) -ocotillol type ginsenoside glycine derivatives of the formula
Chemical name: (20S,24R) -3-O-aminoacetyl-dammar-20, 24-epoxy-3 β,6 α,12 β, 25-tetraol;
the molecular formula is as follows: C32H55NO 6;
molecular weight: 549.4, respectively;
white powder, easily soluble in methanol, ethanol, and pyridine. TLC detection, 10% H2SO4 ethanol solution as developer, heating at 105 deg.C to purple.
The preparation method of the (20S,24R) -ocotillol ginsenoside glycine derivative comprises the following steps:
(1) adding appropriate amount of Ocotillol, BOC-Gly and DMAP into THF, and stirring uniformly;
(2) sequentially adding EDCI and DIPEA into the mixed solution, reacting at room temperature, and monitoring the reaction process by TLC;
(3) adding ethyl acetate into the reaction product to dilute the reaction product in a proper amount;
(4) extracting the diluent by ethyl acetate, combining organic phases, and sequentially washing by using a proper amount of water and a proper amount of saturated saline solution;
(5) passing the washing liquid over anhydrous Na2SO4Drying, filtering, concentrating, and purifying by column chromatography to obtain intermediate compound;
(6) the intermediate was dissolved in dichloromethane, and TFA was added dropwise thereto, and after reaction at room temperature, saturated NaHCO was slowly added to the reaction system3The solution adjusts the solution pH to neutral.
(7) Extracting the neutral liquid by using a proper amount of dichloromethane, combining organic phases, and sequentially washing by using a proper amount of water and a proper amount of saturated saline solution;
(8) drying the washing solution by using anhydrous Na2SO4, and purifying by using column chromatography to obtain the (20S,24R) -ocotillol type ginsenoside glycine derivative.
The mass ratio of the Ocotillol, BOC-Gly and DMAP added in the step (1) is 1:1.5:1.5, the THF is 20-50ml, the stirring is carried out for 30min, the mass ratio of the EDCI and the DIPEA added in the step (2) is 1:1, and the reaction time at room temperature is 48 h.
The volume ratio of the ethyl acetate added in the step (3) to the THF in the step (1) is 4:5, and the volume ratio of the ethyl acetate added in the step (4) to the THF in the step (1) is 6: 5. The volume ratio of water to saturated saline is 1:1.
The column chromatography purification used in the step (5) is eluent: n-hexane: ethyl acetate 5:1
The room-temperature reaction in the step (6) is 2 hours.
The volume ratio of the dichloromethane in the step (7) to the THF in the step (1) is 5: 6. The volume ratio of water to saturated saline solution is 1:1.
The column chromatography purification used in the step (8) is an eluent: dichloromethane: methanol-40: 1.
In step (1), Ocotillol (1.0g,2.0mmol,1.0eq), BOC-Gly (525.5mg,3.0mmol,1.5eq) and DMAP (366.6mg,3.0mmol,1.5eq) were added to THF (25 ml); EDCI (0.7ml,4.0mmol,2.0eq) and DIPEA (0.66ml,4.0mmol,2.0eq) were added in step (2); the ethyl acetate in the step (3) is 20 ml; the ethyl acetate in the step (4) is 30ml, and 30ml of water and 30ml of saturated saline are sequentially used; 5ml of the dichloromethane in the step (6), and 2.1ml of TFA2 is added; 30ml of methylene chloride in the step (7) is sequentially added with 30ml of water and 30ml of saturated saline.
The invention relates to application of (20S,24R) -ocotillol type ginsenoside glycine derivatives in preparing medicines for treating neutrophilic asthma.
When the present invention is used for preparing a drug for treating hormone-resistant asthma, the drug is safe for oral administration or parenteral administration. In the case of oral administration, it may be administered in any conventional form, such as powder, granule, tablet, capsule, pill, drop pill, soft capsule, floating agent, oral liquid, suspension, syrup, buccal tablet, spray or aerosol, etc.; when the drug is administered parenterally, it may take any conventional form, such as injections: such as intravenous injection, ointment, suppository, transdermal administration, inhalant, etc.
The present invention relates to a pharmaceutical composition for treating hormone-resistant asthma, which comprises a single active ingredient or a combination of an active ingredient and a solid or liquid excipient, wherein the solid or liquid excipient is well known in the art, and examples of the solid excipient include lactose, starch, dextrin, calcium carbonate, synthetic or natural aluminum sulfate, magnesium chloride, magnesium stearate, sodium bicarbonate, dried yeast and other liquid excipients such as water, glycerin, propylene glycol, simple syrup, ethanol, ethylene glycol, polyethylene glycol, sorbitol and other ointments.
No (20S,24R) -ocotillol ginsenoside glycine derivatives are found: the report of the preparation method and the pharmaceutical application of (20S,24R) -3-O-aminoacetyl-dammar-20, 24-epoxy-3 beta, 6 alpha, 12 beta, 25-tetrol, the compound belongs to a new compound and is obtained by first synthesis; the new medicinal application of the compound in treating the hormone-resistant asthma disease is discovered for the first time. Animal model experiments are carried out by adopting the (20S,24R) -ocotillol ginsenoside glycine derivative, and experimental results prove that the (20S,24R) -ocotillol ginsenoside glycine derivative has a protective effect on lung function of mice of experimental models of NTHi infection induced hormone-resistant asthma; the inhibitor has an inhibiting effect on the expression of proinflammatory factor THF-alpha in serum of a mouse infected and induced by NTHi and the expression of neutrophil-activating factor IL-8, so that the airway neutrophilic granulocyte inflammation is reduced; has reversing effect on lung pathological changes of NTHi infection induced hormone-resistant asthma mice.
The invention has the beneficial effects that the new compound (20S,24R) -ocotillol type ginsenoside glycine derivative. The new and synthesized ginsenoside derivative monomer is used for treating the hormone-resistant asthma for the first time, has the characteristic of remarkable curative effect, and the dosage of the effective substance can be changed according to the administration mode, the age and the weight of a patient, the severity of the disease condition and other similar factors.
Drawings
FIG. 1 is a drawing of a compound of the present invention1H-NMR spectrum;
FIG. 2 is a drawing of a compound of the present invention13A C-NMR spectrum;
figure 3(a) is a graph of the effect of a compound of the invention on expiratory volume in lung function in model mice, (n ═ 10, mean ± SD);
fig. 3(b) is a graph of the effect of compounds of the present invention on lung function residual capacity in model mouse lung function, (n ═ 10, mean ± SD);
figure 3(c) is a graph of the effect of compounds of the invention on airway resistance in lung function in model mice, (n ═ 10, mean ± SD);
FIG. 4(a) is a graph showing the effect of the compound of the present invention on the level of IL-8 in serum of a model mouse (n 10, mean. + -. SD)
FIG. 4(b) is a graph showing the effect of the compound of the present invention on the TNF-. alpha.content in serum of model mice (n 10, mean. + -. SD)
Detailed Description
A method for preparing (20S,24R) -ocotillol type ginsenoside glycine derivatives comprises the following steps:
(1) adding appropriate amount of Ocotillol, BOC-Gly and DMAP into THF, and stirring;
(2) adding EDCI and DIPEA into the mixed solution in sequence, reacting at room temperature, and monitoring the reaction process by TLC;
(3) adding ethyl acetate into the reaction for dilution;
(4) extracting the diluent by ethyl acetate, combining organic phases, and sequentially washing by using a proper amount of water and a proper amount of saturated saline solution;
(5) drying the washing liquid by using anhydrous Na2SO4, filtering, concentrating, and purifying eluent by using column chromatography to obtain an intermediate compound;
(6) dissolving the intermediate in dichloromethane, dropwise adding TFA, reacting at room temperature for 2 hours, and slowly adding saturated NaHCO3 solution into the reaction system to adjust the pH of the solution to be neutral;
(7) extracting the neutral liquid by dichloromethane, combining organic phases, and washing by water and saturated saline solution in sequence;
(8) drying the washing solution by using anhydrous Na2SO4, and purifying eluent by using column chromatography to obtain the (20S,24R) -ocotillol type ginsenoside glycine derivative.
The mass ratio of the Ocotillol, BOC-Gly and DMAP added in the step (1) is 1:1.5:1.5, the THF is 20-50ml, and the stirring is 30min
The mass ratio of EDCI to DIPEA added in step (2) was 1:1, and the reaction time was 48h at room temperature.
The volume ratio of the ethyl acetate added in the step (3) to the THF in the step (1) is 4: 5.
The volume ratio of the ethyl acetate added in the step (4) to the THF in the step (1) is 6: 5. The volume ratio of water to saturated saline is 1:1.
The column chromatography purification used in step (5) is eluent: n-hexane: ethyl acetate 5:1.
The room-temperature reaction in the step (6) is 2 hours.
The volume ratio of the dichloromethane in the step (7) to the THF in the step (1) is 5: 6. The volume ratio of water to saturated saline solution is 1:1.
The column chromatography purification used in step (8) is an eluent: dichloromethane: methanol-40: 1.
In step (1), Ocotillol (1.0g,2.0mmol,1.0eq), BOC-Gly (525.5mg,3.0mmol,1.5eq) and DMAP (366.6mg,3.0mmol,1.5eq) were added to THF (25 ml); EDCI (0.7ml,4.0mmol,2.0eq) and DIPEA (0.66ml,4.0mmol,2.0eq) were added in step (2); the ethyl acetate in the step (3) is 20 ml; the ethyl acetate in the step (4) is 30ml, and 30ml of water and 30ml of saturated saline are sequentially used; 5ml of the dichloromethane in the step (6), and 2.1ml of TFA2 is added; 30ml of methylene chloride in the step (7) is sequentially added with 30ml of water and 30ml of saturated saline.
Example 1
A method for preparing (20S,24R) -Ocotillol type ginsenoside glycine derivatives by using Ocotillol as a prototype comprises the following steps:
add Ocotillol (1.0g,2.0mmol,1.0eq), BOC-Gly (525.5mg,3.0mmol,1.5eq) and DMAP (366.6mg,3.0mmol,1.5eq) to THF (25ml) and stir for 30 min;
adding EDCI (0.7ml,4.0mmol,2.0eq) and DIPEA (0.66ml,4.0mmol,2.0eq) into the mixed solution in sequence, and reacting at room temperature for 48h after the addition is finished;
monitoring the reaction process by TLC, and adding ethyl acetate (20ml) into the reaction system for dilution after the reaction is finished;
the dilution was extracted with ethyl acetate (3X 10ml), the organic phases were combined and washed successively with water (3X 10ml) and saturated brine (3X 10 ml);
the washing liquid is anhydrous Na2SO4Drying, filtering, concentrating, and purifying by column chromatography (eluent: n-hexane: ethyl acetate: 5:1) to obtain an intermediate compound (0.8g, Yield: 61.6%);
the intermediate (0.8g,1.2mmol) was dissolved in dichloromethane (5ml) and TFA (2.1ml) was added dropwise thereto. After the addition was completed, the reaction was carried out at room temperature for 2 hours, and the progress of the reaction was monitored by TLC. When the reaction is finished, slowly adding saturated NaHCO into the reaction system3Adjusting the pH value of the solution to be neutral;
the neutral liquid was extracted with dichloromethane (3X 10 ml). The organic phases were combined and washed successively with water (3X 10ml) and saturated brine (3X 10 ml);
passing the washing solution through anhydrous Na2SO4Drying, filtering, concentrating and purifying by column chromatography (eluent: dichloromethane: methanol: 40:1) to obtain the (20S,24R) -ocotillol type ginsenoside glycine derivative.
Structure identification is carried out by adopting a spectroscopy method:
compound (I)
HR-ESI-MS spectrum gives molecular weight of 549.4, and combined with 1H NMR and 13C NMR spectra to determine the molecule
The formula is C32H55NO 6. 1H-NMR spectra of Compound
δ4.52-4.50(m,1H),4.13-4.08(m,1H),3.86-3.83(m,1H),3.53-3.48(m,1H),3.46(s,2H),2.21-2.17(m,1H),2.05-1.96(m,3H),1.93-1.81(m,6H),1.70-1.54(m,8H),1.27(s,3H),1.26(s,3H),1.16(s,3H),1.09(s,3H),1.06(s,3H),1.04(s,3H),0.98(s,1H),0.97(s,1H),0.93(s,3H),0.93(s,3H).
13C-NMR spectra of Compound
δ173.40(C＝O),86.48(20-C),85.43(24-C),81.62(3-C),70.80(12-C),70.14(25-C),68.30(6-C),61.29(5-C),51.82(14-C),49.89(9-C),49.00(13-C),47.88(17-C),47.03(7-C),43.73(NH-C-C＝O),40.87(8-C),38.94(4-C),38.38(1、10-C),32.60(15-C),31.25(11-C),31.19(22-C),30.70(28-C),28.58(23-C),27.90(27-C),27.52(26-C),26.13(21-C),24.95(16-C),23.39(2-C),18.12(30-C),17.49(19-C),16.93(18-C),16.53(29-C)
And comprehensively analyzing to identify the compound as (20S,24R) -3-O-aminoacetyl-dammar-20, 24-epoxy-3 beta, 6 alpha, 12 beta, 25-tetrol. The SciFinder search does not find a literature report about the compound, and the compound is a new compound.
Of the compound1H-NMR spectrum,13The C-NMR spectrum is shown in the attached figures 1 and 2 respectively.
The effects of the present invention are further illustrated by the pharmacodynamic test examples below.
Test example 1: effect of (20S,24R) -ocotillol type ginsenoside glycine derivatives on pulmonary function of mice in a hormone-resistant asthma model.
Adopting a non-typable haemophilus influenzae (NTHi) infection method to induce chicken Ovalbumin (OVA) atomization to excite an asthma mouse to establish a neutrophilic granulocyte asthma model. 60 mice were randomly divided into a normal control group, a model group, a compound ((20S,24R) -ocotillol type ginsenoside glycine derivative) low and high dose group (10 mg/kg; 20mg/kg), a treatment control group (dexamethasone), and a combination treatment group (dexamethasone + compound high dose). Mice were injected intraperitoneally with 200. mu.L of sensitizing solution (200. mu.L of physiological saline containing 100. mu.g of OVA and 2mg of aluminum hydroxide gel) on days 0, 7 and 14, respectively, from the start of the experiment. From the 21 st day of the experiment, mice were given 10mL of 5% OVA solution nebulization by ultrasonic nebulizer for 4 consecutive days with daily nebulization for 30 min. Mice were given tracheal inoculation with NTHi on day 25 of the experiment, i.e. 24h after the last OVA nebulization. After inoculating bacteria for 1h, respectively intragastrically administering normal control group and model group with normal saline, respectively intragastrically administering compound dosage groups, and continuously administering for 3 days; at the same time, 100 mu L dexamethasone solution is injected into the abdominal cavity of the mice of the treatment control group and the combination treatment group, the administration dose is 2mg/kg, the administration is continuously carried out for 3 days, and the other groups are respectively administered with sterile normal saline with equal dose.
After the mice were weighed, the mice were anesthetized by intraperitoneal injection of 1% pentobarbital sodium injection according to body weight. After deep anesthesia, the mouse is fixed on a mouse board in a supine position, after an trachea is separated, a transverse incision is cut at the upper end of a tracheal cartilage ring, a tracheal cannula is inserted and fixed by an operation line, and the other end of the tracheal cannula is connected with an airflow sensor, a pressure sensor and a built-in small animal respirator of the Buxco small animal lung function instrument. Connecting a PFT detection system of the pulmonary function instrument, setting a standard value, tracing a section of calm breath, and detecting the 0.1 second forced expiratory volume (FEV0.1), the pulmonary Function Residual Capacity (FRC) and the airway Resistance (RI) after the breath is stable.
As a result: compared with the normal control group, the lung function parameters of the mice in the model group are all obviously reduced, and the differences have statistical significance: (###P<0.001 or##P<0.01), indicating that the molding is successful. Compound high dose group, treatment control group and combination compared with model groupThe lung function parameters of the treatment groups are obviously increased, and the difference has statistical significance (P)<0.05 or P<0.01), the results are shown in FIGS. 3(a) - (c), Table 1.
Table 1 effect of (20S,24R) -ocotillol type ginsenoside glycine derivatives and dexamethasone on lung function in model mice (n ═ 10, mean ± SD)
And (4) conclusion: the (20S,24R) -ocotillol type ginsenoside glycine derivative can improve the lung function of a mouse model with hormone-resistant asthma, has the effect similar to that of dexamethasone, and can remarkably enhance the effect of the dexamethasone on improving the lung function of the mouse model with neutrophilic granulocyte asthma.
Test example 2: observing the influence of the (20S,24R) -ocotillol type ginsenoside glycine derivative on the expression of proinflammatory factor TNF-alpha and neutrophil chemotactic factor IL-8 in the serum of a mouse model of neutrophilic asthma.
The molding method and the administration method were the same as in test example 1.
After the test of the lung function is finished, blood is taken from eyeballs of the mice, the blood is placed on ice, after all blood samples of the experimental animals are collected, the blood is agglutinated for 30min at room temperature, the blood samples are placed overnight at 4 ℃ to completely release cytokines, the blood samples are centrifuged for 10min at 3000rpm on the 2 nd day at room temperature, upper layer serum is collected and frozen at the temperature of-20 ℃, and the cytokines THF-alpha and IL-8 are detected by adopting an enzyme-linked immunosorbent assay. IL-8 detection kits and TNF-alpha detection kits were purchased from R & D, USA.
As a result: the IL-8 and TNF-alpha contents in the model group are obviously increased compared with the normal group (##P<0.01 or###P<0.001), (20S,24R) -ocotillol type ginsenoside glycine derivative content of IL-8 and TNF-alpha in the high-dose group is remarkably reduced compared with that in the model group*P<0.05 or**P<0.01); the IL-8 and TNF-alpha content of the treatment control group is obviously reduced compared with that of the model group (*P<0.05 or**P<0.01); the IL-8 and TNF-alpha content in the combined treatment group is obviously reduced compared with that in the model group (*P<0.05 or**P<0.01); see FIGS. 4(a) - (b), Table 2.
TABLE 2 Effect of (20S,24R) -ocotillol-type ginsenoside glycine derivatives and dexamethasone on IL-8 and TNF- α levels in serum of model mice (n 10, mean + -SD)
| Group of | IL-8(pg/mL) | TNF-α(pg/mL) |
| Normal group | 1102.68±68.44 | 3.03±0.39 |
| Model set | 2177.27±198.5### | 8.06±1.1### |
| Compound low dose group | 1974.71±100.49* | 7.02±1.08 |
| High dose group of compounds | 1823.94±157.68** | 6.22±1.83* |
| Treatment control group | 1802.59±160.21** | 6.44±1.5* |
| AssociationTreatment group | 1675.02±193.59** | 5.09±0.76** |
And (4) conclusion: IL-8 and TNF-alpha have inhibition effect on the expression of proinflammatory factor TNF-alpha in the serum of a neutrophilic granulocyte asthma model mouse, and have inhibition effect on the expression of neutrophilic granulocyte chemotactic factor IL-8, so that the neutrophilic granulocyte inflammation of an airway is reduced.
Test example 3: the (20S,24R) -ocotillol type ginsenoside glycine derivatives have the reversing effect on the lung pathological changes of mice in the model of neutrophilic asthma.
The molding method and the administration method were the same as in test example 1.
And (3) perfusing and fixing each experimental mouse by using a formaldehyde solution, and observing pathological changes of lung tissues by hematoxylin-eosin staining.
As a result: the bronchus and alveolus of the normal group of mice are complete in structure, and inflammatory cell infiltration is not seen in lung tissues; the lung tissue of the model group mouse can show obvious neutrophilic granulocyte inflammation, and the airway, the blood vessel and the lung parenchyma have a large amount of inflammatory cell infiltration such as neutrophilic granulocyte, lymphocyte and the like, so that part of alveoli are blocked, and the alveoli interval is widened. (20S,24R) -ocotillol type ginsenoside glycine derivatives are administered to mouse lung tissues of a high-dose group and a dexamethasone group, and neutral granulocyte infiltration can be seen, a small part of alveoli is blocked, and the alveoli interval is widened; in the mice of the combination treatment group, mild neutrophil infiltration, slight alveolar occlusion and alveolar septal broadening can be seen in lung tissues.
And (4) conclusion: the (20S,24R) -ocotillol ginsenoside glycine derivative has reversal effect on mouse lung pathological changes of a neutrophilic granulocyte asthma model induced by NTHi infection, and can relieve disease symptoms of neutrophilic granulocyte inflammation.
Claims (10)
1. A (20S,24R) -ocotillol ginsenoside glycine derivative, characterized by the formula:
chemical name: (20S,24R) -3-O-aminoacetyl-dammar-20, 24-epoxy-3 β,6 α,12 β, 25-tetraol;
the molecular formula is as follows: c32H55NO6；
Molecular weight: 549.4, respectively;
white powder, easily soluble in methanol, ethanol, pyridine; TLC detection, 10% H2SO4The ethanol solution is a color developing agent, and is heated to be purple at 105 ℃.
2. A process for the preparation of a (20S,24R) -ocotillol ginsenoside glycine derivative of claim 1, comprising the steps of:
(1) adding appropriate amount of Ocotillol, BOC-Gly and DMAP into THF, and stirring uniformly;
(2) sequentially adding EDCI and DIPEA into the mixed solution, reacting at room temperature, and monitoring the reaction process by TLC;
(3) adding ethyl acetate into the reaction product to dilute the reaction product in a proper amount;
(4) extracting the diluent by ethyl acetate, combining organic phases, and sequentially washing by using a proper amount of water and a proper amount of saturated saline solution;
(5) passing the washing liquid over anhydrous Na2SO4Drying, filtering, concentrating, and purifying by column chromatography to obtain intermediate compound;
(6) the intermediate was dissolved in dichloromethane, and TFA was added dropwise thereto, and after reaction at room temperature, saturated NaHCO was slowly added to the reaction system3The solution adjusts the solution pH to neutral.
(7) Extracting the neutral liquid by using a proper amount of dichloromethane, combining organic phases, and sequentially washing by using a proper amount of water and a proper amount of saturated saline solution;
(8) drying the washing solution by using anhydrous Na2SO4, and purifying by using column chromatography to obtain the (20S,24R) -ocotillol type ginsenoside glycine derivative.
3. The method for preparing a ginsenoside glycine derivative of (20S,24R) -Ocotillol type according to claim 2, wherein the amount of the material of the Ocotillol, BOC-Gly and DMAP added in the step (1) is 1:1.5:1.5, the THF is 20-50ml, the stirring is 30min, the amount of the material of the EDCI and DIPEA added in the step (2) is 1:1, and the reaction time at room temperature is 48 h.
4. The method for preparing a (20S,24R) -ocotillol type ginsenoside glycine derivative according to claim 2, wherein the volume ratio of the ethyl acetate added in the step (3) to the THF in the step (1) is 4:5, and the volume ratio of the ethyl acetate added in the step (4) to the THF in the step (1) is 6: 5. The volume ratio of water to saturated saline is 1:1.
5. The process for preparing (20S,24R) -ocotillol type ginsenoside glycine derivatives of claim 2, wherein the column chromatography used in step (5) is used for purifying the eluent: n-hexane: ethyl acetate 5:1
6. The process for preparing (20S,24R) -ocotillol type ginsenoside glycine derivatives of claim 2, wherein the reaction at room temperature in the step (6) is 2 h.
7. The process for preparing a (20S,24R) -ocotillol ginsenoside glycine derivative of claim 2, wherein the volume ratio of the dichloromethane in the step (7) to the THF in the step (1) is 5: 6. The volume ratio of water to saturated saline solution is 1:1.
8. The process for preparing a (20S,24R) -ocotillol ginsenoside glycine derivative of claim 2, wherein the column chromatography used in step (8) is used for purifying as an eluent: dichloromethane: methanol-40: 1.
9. The process for preparing a (20S,24R) -Ocotillol ginsenoside glycine derivative of claim 2, wherein 1.0g,2.0mmol,1.0eq of Ocotillol, 525.5mg,3.0mmol,1.5eq of BOC-Gly and 366.6mg,3.0mmol,1.5eq of DMAP are added to 25ml of THF in step (1); adding 0.7ml,4.0mmol,2.0eq EDCI and 0.66ml,4.0mmol,2.0eq DIPEA into the step (2); the ethyl acetate in the step (3) is 20 ml; the ethyl acetate in the step (4) is 30ml, and 30ml of water and 30ml of saturated saline are sequentially used; 5ml of the dichloromethane in the step (6), and 2.1ml of TFA2 is added; 30ml of methylene chloride in the step (7) is sequentially added with 30ml of water and 30ml of saturated saline.
10. Use of a (20S,24R) -ocotillol-type ginsenoside glycine derivative of claim 1 as a prophylactic and therapeutic agent for hormone-resistant asthmatic diseases.
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| CN102875628A (en) * | 2012-10-30 | 2013-01-16 | 中国药科大学 | (20S,24S)-ocotillol ginsenoside derivatives with antibacterial activity, and preparation method and application thereof |
| CN102924556A (en) * | 2012-11-05 | 2013-02-13 | 烟台大学 | (20S, 24R)-ocotillol type ginsenoside derivative having antibacterial activity and preparation method and application thereof |
| CN106967143A (en) * | 2017-05-01 | 2017-07-21 | 烟台大学 | Novel pyxinol derivatives of structure and its production and use |
| CN109776647A (en) * | 2019-02-14 | 2019-05-21 | 烟台大学 | Pyxinol esterification derivative with anti-inflammatory activity and its preparation method and application |
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| CN102875628A (en) * | 2012-10-30 | 2013-01-16 | 中国药科大学 | (20S,24S)-ocotillol ginsenoside derivatives with antibacterial activity, and preparation method and application thereof |
| CN102924556A (en) * | 2012-11-05 | 2013-02-13 | 烟台大学 | (20S, 24R)-ocotillol type ginsenoside derivative having antibacterial activity and preparation method and application thereof |
| CN106967143A (en) * | 2017-05-01 | 2017-07-21 | 烟台大学 | Novel pyxinol derivatives of structure and its production and use |
| CN109776647A (en) * | 2019-02-14 | 2019-05-21 | 烟台大学 | Pyxinol esterification derivative with anti-inflammatory activity and its preparation method and application |
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