CN115197191A - Novel traditional Chinese medicine flavone derivative and preparation method and application thereof - Google Patents
Novel traditional Chinese medicine flavone derivative and preparation method and application thereof Download PDFInfo
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- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
- C07D311/30—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only not hydrogenated in the hetero ring, e.g. flavones
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Abstract
The invention discloses a novel traditional Chinese medicine flavone derivative and a preparation method and application thereof. The novel traditional Chinese medicine flavone derivative has a structure with the following general formula:
wherein R is 1 、R 2 Selected from fluorine-containing alkyl group consisting of 1-5 carbon atoms, cycloalkyl or cycloalkylmethyl group consisting of 3-6 carbon atoms, or saturated or unsaturated alkyl group of C2-C6; r 3 、R 7 Selected from hydrogen, or alkoxy; r 4 、R 5 Selected from hydrogen, or methyl; r 6 、R 8 Selected from hydrogen, hydroxy, or alkoxy. The novel Chinese medicinal flavone derivatives have effects of inhibiting phosphodiesterase, especially PDE4 activity, and effectively inhibiting TNF-alpha release, neutrophil or eosinophil increase, and protecting nerve cells, myocardial cells and vascular endothelial cells. The novel traditional Chinese medicine flavone derivative has good safety, so that the novel traditional Chinese medicine flavone derivative has important development significance.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a novel traditional Chinese medicine flavone derivative as well as a preparation method and application thereof.
Background
Phosphodiesterases are a member of the enzyme family, and 11 PDE enzyme families (PDE 1-PDE 11) are known to date, which differ in their substrate specificity (cAMP, cGMP or both) and their dependence on other substrates (e.g. calmodulin). Inhibition of different types of PDE isozymes leads to intracellular accumulation of cAMP and/or cGMP, which can be useful in the treatment of different inflammation-related diseases. PDE4 is mainly distributed in various inflammatory cells, such as mast cells, macrophages, eosinophils, lymphocytes and epithelial cells, and can increase intracellular concentration by inhibiting enzyme activity, which helps to reduce the harm of inflammatory reaction to the body. The major PDE isozymes are also type 4 in cells important for allergic inflammation (lymphocytes, mast cells, eosinophils, macrophages). Therefore, inhibition of PDE4 with suitable inhibitors is considered an important starting point for the treatment of various allergy-induced diseases. PDE4 inhibitors have been developed to date as anti-inflammatory drugs, such as roflumilast, primarily for the treatment of inflammation of the lung, particularly asthma and chronic obstructive pulmonary disease; difamilast is used for the treatment of atopic dermatitis; apremilast, in turn, is used in the treatment of psoriatic arthritis; meanwhile, inhibitors of PDE1, PDE3 and PDE5 have also been used in the treatment of clinical cardiovascular and cerebrovascular diseases, such as vinpocetine, dipyridamole, milrinone, sildenafil, etc.
Meanwhile, an important feature of PDE4 inhibitors is the inhibition of the release of tumor necrosis factor (TNF-. Alpha.) from inflammatory cells. TNF- α is an important proinflammatory cytokine affecting a variety of biological processes, which can be released from activated local cells, activated T lymphocytes, mast cells, basophils, fibroblasts, endothelial cells, and astrocytes in the brain. TNF- α itself has an activating effect on neutrophils, eosinophils, fibroblasts and endothelial cells, which in turn release different tissue-destructive mediators. TNF- α causes increased levels of other proinflammatory cytokines such as GM-CSF (granulocyte-macrophage colony stimulating factor) or interleukin-8 in monocytes, macrophages and T lymphocytes. TNF- α plays a key role in a variety of diseases such as respiratory inflammation, joint inflammation, endotoxic shock, tissue rejection, AIDS and a variety of other immune disorders, as TNF- α promotes inflammation and catabolic action. Therefore, PDE4 inhibitors are also useful in the treatment of TNF- α related diseases.
Although PDE4 inhibitors are known to have shown beneficial pharmacological effects, such inhibitors have adverse effects that cause diarrhea, nausea, and the like. Therefore, the research of novel specific inhibitors to overcome these adverse effects becomes one of the hot spots of inhibitor drug research. The natural traditional Chinese medicine derivative is a treasure house discovered by a lead compound for developing new medicines, a novel inhibitor is searched from the natural traditional Chinese medicine derivative, and the natural traditional Chinese medicine derivative has important significance for developing anti-inflammatory medicines of the inhibitor with high curative effect and small side effect.
Traditional Chinese medicine flavonoids have wide pharmacological effects, flavones are found in medicinal plants as effective components, and at present, in clinical medicines, many medicines containing flavones, especially in traditional Chinese medicine preparations, flavones play main roles as important components, for example: shuanghuanglian injection and the like. Ginkgo contains dozens of flavonoid compounds including isoflavone, flavonol, flavone, flavanone, etc., and is mainly used for treating hypertension and reducing blood lipid; however, the traditional flavone compound has the problems of difficult water solubility, easy metabolism in vivo and low bioavailability, thereby greatly limiting the better development of the traditional flavone compound in the aspect of clinical application.
Disclosure of Invention
In order to overcome the problems in the background art, the invention provides a novel traditional Chinese medicine flavone derivative and a preparation method and application thereof. Pharmacological experiments prove that the novel traditional Chinese medicine flavone derivatives have the effects of inhibiting phosphodiesterase activity and protecting nerve cells, myocardial cells and vascular endothelial cells.
In order to realize the purpose of the invention, the technical scheme of the invention is as follows:
the invention provides a novel traditional Chinese medicine flavone derivative, which has a structure shown in the following general formula:
wherein R is 1 Selected from fluorine-containing alkyl consisting of 1 to 5 carbon atoms, cycloalkyl or cycloalkylmethyl consisting of 3 to 6 carbon atoms or saturated or unsaturated alkyl of C2 to C6; r is 2 Selected from hydrogen or fluorine-containing alkyl group consisting of 1 to 5 carbon atoms, cycloalkyl or cycloalkylmethyl group consisting of 3 to 6 carbon atoms, or saturated or unsaturated alkyl group of C2 to C6; r 3 Selected from hydrogen or alkoxy; r 4 Selected from hydrogen or alkyl; r 5 Selected from hydrogen or alkyl; r 6 Selected from hydrogen, hydroxy or alkoxy; r is 7 Selected from hydrogen or alkoxy; r is 8 Selected from hydrogen, hydroxyl or alkoxy.
Further: the novel traditional Chinese medicine flavone derivative has a structure of the following general formula:
wherein R is 1 Is selected from-CH 3 、
Or
R 2 Is selected from-CH 3 、
Or alternatively
R 3 Is selected from-H or-OCH 3 ；R 4 Is selected from-H or-CH 3 ；R 5 Is selected from-H or-CH 3 ；R 6 Is selected from-H or-OH or-OCH 3 ；R 7 Is selected from-H or-OCH 3 ；R 8 Is selected from-H. -OH or-OCH 3 。
Further: the novel traditional Chinese medicine flavone derivatives are MX1, MX2, MX4, HHQ2, YD2, SFH, YD3, HHQ3, YM2, HP3, HP4, SHT2, SHT3, SCJ2 and SCJ3, and specifically comprise the following structures:
the invention also provides a preparation method of the novel traditional Chinese medicine flavone derivative, which comprises the following steps:
mixing a derivative X of 2-methoxyacetophenone protected by MOM hydroxyl with a derivative Y of benzaldehyde, carrying out nucleophilic addition under the catalysis of alkali, generating chalcone by a claisen-Schmitt method, carrying out intramolecular cyclization, and removing a protective group under an acidic condition to obtain novel traditional Chinese medicine flavone derivatives MX1, MX2, MX4, HHQ2, YD2, SFH, YD3, SMX1, SHQ2, HHQ3 and YM2;
or mixing MOM hydroxyl protected 2-hydroxyacetophenone derivative Z with benzaldehyde derivative Y, adding pyrrolidine to react completely, removing protecting groups, extracting, washing, drying, concentrating, and purifying to obtain new traditional Chinese medicine flavone derivatives HP2, HP3, HP4, SHT2, SHT3, SHP3, SHT5, SCJ2 and SCJ3;
the structure of the derivative X is as follows:
the structure of the derivative Y is as follows:
the structure of the derivative Z is as follows:
wherein R is 1 Is selected from-CH 3 、
Or
R 2 Is selected from-CH 3 、
Or
R 3 Is selected from-H or-OCH 3 ；R 4 Is selected from-H or-CH 3 ；R 5 Is selected from-H or-CH 3 ；R 6 Is selected from-H or-OH or-OCH 3 ；R 7 Is selected from-H or-OCH 3 。
Further: the novel traditional Chinese medicine flavone derivative also comprises pharmaceutically acceptable salts thereof, and can be obtained by neutralizing acid with inorganic base or organic base, or neutralizing base with inorganic acid or organic acid by a conventional method.
And further: the novel traditional Chinese medicine flavone derivatives also comprise D type, L type or D, L-mixture thereof.
Further: the novel traditional Chinese medicine flavone derivative also comprises diastereoisomers thereof.
The invention also provides application of the novel traditional Chinese medicine flavone derivative in preparation of phosphodiesterase PDE and/or TNF-alpha inhibitors.
Further: the novel traditional Chinese medicine flavone derivative can effectively inhibit phosphodiesterase PDE and TNF-alpha.
The invention also provides application of the novel traditional Chinese medicine flavone derivative in preparing a medicine for preventing and treating inflammatory and/or allergic diseases.
Further, the novel Chinese medicinal flavone derivatives can relieve the symptoms of inflammatory and/or allergic diseases by effectively inhibiting the activity of phosphodiesterase PDE, inhibiting the release of TNF-alpha and inhibiting the increase of neutrophils and eosinophils.
Further: the phosphodiesterase is PDE4, PDE2, PDE3, PDE5 and PDE 10.
Preferably: the phosphodiesterase is PDE4.
Further: inflammatory diseases associated with inhibition of TNF- α release include arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis, gastritis, gastric ulceration, osteoporosis, sepsis, septic shock, gram negative sepsis, toxic shock syndrome, respiratory distress syndrome, asthma and other chronic lung diseases, bone resorption diseases or graft rejection or other autoimmunity, lupus erythematosus, multiple sclerosis, glomerulonephritis and uveitis, insulin dependent diabetes mellitus and chronic demyelination.
Further: inflammatory or allergic diseases associated with eosinophilia include bronchial asthma, allergic rhinitis, allergic conjunctivitis, atopic dermatitis, eczema, allergic vasculitis, eosinophil-induced inflammation, such as eosinophilic fasciitis, eosinophilic pneumonia and PIE syndrome (eosinophilic lung infiltration), urticaria, ulcerative conjunctivitis, crohn's disease, psoriasis and keratosis.
And further: the compounds of the invention are also useful in the treatment of infections associated with TNF- α, including viral and parasitic infections, malaria, leishmaniasis, fever due to infection, muscle pain due to infection, AIDS, and cachexia.
Further: diseases associated with increased neutrophils include chronic obstructive pulmonary disease.
The invention also provides application of the novel traditional Chinese medicine flavone derivative in preparing a medicine for preventing and treating diseases of a nervous system and/or a cardiovascular system.
Further: the novel traditional Chinese medicine flavone derivative can effectively inhibit Abeta (beta-amyloid) aggregates and/or CoCl 2 Induced nerve cell and/or myocardial cell damage, or effective reversal of oxidized low density lipidsVascular endothelial cell injury induced by the protein ox-LDL, thereby achieving the effect of protecting nerve cells or cardiovascular cells.
Further: the nervous system and/or cardiovascular system diseases include senile dementia (Alzheimer's disease), memory loss, parkinson's disease, depressive anxiety disorder, schizophrenia, stroke, intermittent claudication, cardiovascular injury, arteriosclerosis, hyperlipidemia, hyperglycemia, cerebral ischemic injury diseases, benign prostatic hyperplasia, pollakiuria, nocturia, and incontinence, urinary calculus-induced striatal pain and sexual dysfunction.
And further: the medicine is in the form of tablet, oral liquid, aerosol, pill, capsule, granule, paste, drop pill, syrup, powder, granule, tincture, powder for injection or injection.
Further: the medicament is administered orally, parenterally, intravenously, transdermally, topically, by inhalation, and intranasally.
Further: the dose of the drug is administered as a single dose once a day, or is divided into two or more doses each of 0.001-100mg per day.
Further: the medicament also comprises at least one of an adjuvant, a carrier and an additive.
And further: the carrier comprises at least one of calcium carbonate, calcium phosphate, sodium phosphate, lactose, starch, mannitol, alginate, gelatin, guar gum, magnesium stearate, aluminum stearate, methyl cellulose, talc, highly dispersed silica, silicone oil, stearic acid, gelatin, agar, vegetable or animal fats and oils, polyethylene glycol.
And further: the adjuvants include sweetening agents, flavoring agents, preservatives, stabilizers, wetting agents, osmotic agents, emulsifiers, coating agents, cosolvents, at least one of salts for controlling osmotic pressure or for buffering, sugars or sugar alcohols and/or viscosity modifiers.
Further: the additive comprises at least one of tartrate and citrate buffer, ethanol and complexing agent.
Further: for viscosity control, the additives can also use liquid polyethylene oxide, microcrystalline cellulose, polyvinylpyrrolidone, dextran or gelatin.
Further: oil suspensions for parenteral or topical application may contain synthetic or semi-synthetic oils of vegetable origin, including liquid fatty acid esters thereof having from 8 to 22 carbon atoms in the fatty acid chain, including palmitic, lauric, tridecanoic, margaric, stearic, eicosanoic, myristic, behenic, pentadecanoic, linoleic, elaidic, basilic, erucic or oleic acid, which are esterified with mono-and trihydric alcohols having from 1 to 6 carbon atoms, including methanol, ethanol, propanol, butanol, pentanol or isomers thereof, ethylene glycol or glycerol; the fatty acid ester includes Miglyole, isopropyl myristate, isopropyl palmitate, isopropyl stearate, PEG-6 decanoic acid, caprylic/capric acid esters of saturated fatty alcohols, polyoxyethylene glycerol trioleate, ethyl oleate, waxy fatty acid esters such as synthetic duck tail gland fat, isopropyl esters of coconut oil fatty acids, oleyl oleate, decyl oleate, ethyl lactate, dibutyl phthalate, diisopropyl adipate, fatty acid esters of polyhydric alcohols; silicone or fatty alcohols including isotridecanol, 2-octyldodecanol, cetostearyl or oleyl alcohol, oleic acid and vegetable oils including castor oil, almond oil, olive oil, sesame oil, cottonseed oil, peanut oil or soybean oil can also be used.
Further: as solvents, gel formers and cosolvents, water or alcohols are used including ethanol or isopropanol, benzyl alcohol, 2-octyldodecanol, polyethylene glycol, phthalates, adipates, propylene glycol, glycerol, dipropylene glycol, tripropylene glycol, waxes, methyl cellosolve, esters, wulin, dioxane, dimethyl sulfoxide, dimethylformamide, tetrahydroxypyran, cyclohexanone.
And further: as film-forming agents, cellulose ethers which are soluble or swellable in water and organic solvents are used, including hydroxypropylmethylcellulose, methylcellulose, ethylcellulose or soluble starches, sodium carboxymethylcellulose, polyacrylic acid, polyisobutylene acid and salts thereof, sodium pullulan hemiglycolate, alginic acid or propylene glycol alginate vinegar as the sodium salt, gum arabic, xanthan gum, guar gum or carrageenan.
Further: as other adjuvants, glycerol, paraffin of different viscosity, triethanolamine, collagen, and allantoin can also be used.
Further: surfactants, emulsifiers or wetting agents can also be included in the medicament, including sodium lauryl sulfate, fatty alcohol ether sulfates, disodium N-lauryl- β -iminodipropionate, polyethoxylated castor oil or sorbitan oleyl acetate, sorbitan stearate, polysorbate, cetyl alcohol, lecithin, glycerol stearate, polyoxyethylene stearate, alkylphenol polyglycol ethers, ethoxylated cetyl trimethylammonium or alkyl or dialkyl polyglycol ether orthophosphoric acid ethanolamine salts; stabilizers include montmorillonite or colloidal silica; antioxidants including tocopherol or butylated hydroxyanisole; preservatives include parabens.
Compared with the prior art, the invention has the advantages and beneficial effects that:
the invention carries out structural modification on the basis of the original natural structures of the traditional Chinese medicine flavonoid compounds quercetin, luteolin, myricetin, apigenin, wogonin, baicalein, pecan element, marrubin, cyanidin, kaempferol, isorhamnetin, diosmetin, galangin, formononetin and the like, thereby not only enhancing the targeted inhibition activity of the original compounds on PDE, but also obtaining the novel traditional Chinese medicine flavone derivatives with novel structures. Pharmacological experiments prove that the novel traditional Chinese medicine flavone derivatives have good effect of inhibiting the activity of phosphodiesterase, particularly PDE4, and can effectively inhibit the release of TNF-alpha, neutrophilic leukocytosis or eosinophilic cytosis, further relieve the symptoms of inflammatory diseases or allergy, and achieve the effect of preventing and treating the inflammatory diseases or allergy; and also inhibit A beta aggregate-induced nerve cell damage, or CoCl 2 Induced hypoxic injury of nerve cells or cardiac myocytes and effective reversal of ox-LD of vascular endothelial cellsL injury, so as to increase the activity of nerve cells or myocardial cells, avoid the injury of vascular endothelial cells by oxidized low density lipoprotein, and achieve the effect of protecting nerve cells or cardiovascular cells. The novel traditional Chinese medicine flavone derivative disclosed by the invention has a good treatment effect on various diseases, has good safety and has a further development value.
Detailed Description
The technical solution of the present invention is further described in detail with reference to the specific embodiments.
The novel traditional Chinese medicine flavone derivative has the following general formula structure:
wherein R is 1 Is selected from-CH 3 、
Or
R 2 Is selected from-CH 3 、
Or
R 3 、R 7 Is selected from-H or-OCH 3 ；R 4 、R 5 Is selected from-H or-CH 3 ；R 6 、R 8 Is selected from-H or-OH or-OCH 3 。
Example 1: synthesis of novel traditional Chinese medicine flavone derivatives
1. R in the general structure of the synthesis 1 ＝
R 2 ＝-CH 3 、R 3 ＝-H、R 4 ＝-H、R 5 ＝-H、R 6 ＝-H、R 7 ＝-H、R 8 Representative Compound of = -H2- (3-Cyclopropylmethoxy-4-methoxybenzene) -5, 7-dihydroxy-4H-chromen-4-one
Dissolving 1mol of 5, 7-dimethoxymethyl ether (MOM) hydroxyl-protected 2-methoxyacetophenone and an equal amount of 3-cyclopropoxy-4-methoxybenzaldehyde in 100ml of methanol at room temperature, adding 0.3mol of potassium hydroxide, and obtaining 2,4-MOM hydroxyl-protected 4 '-methoxy-5' -cyclopropoxy-6-methoxychalcone by nucleophilic addition and loss of one molecule of water, which is a classical reaction; dissolving 0.65mol of the product in DMSO solution, adding elementary iodine, heating and refluxing for 3-6h, cooling to room temperature, and pouring the reaction into 2% NaHSO 3 In the solution, solid is filtered out after full stirring, then at room temperature, the obtained solid compound is put in methanol and hydrochloric acid (3 mol/L) to remove protective groups under the protection of argon, then cooled to room temperature, decompressed and concentrated to remove methanol, filtered, washed, dried and recrystallized by absolute ethyl alcohol to obtain the representative compound 2- (3-cyclopropylmethoxy-4-methoxybenzene) -5, 7-dihydroxy-4H-chromen-4-one (MX 2). The data for the hydrogen and carbon spectra of nmr are as follows:
compound MX2 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ7.61(s,2H),7.13(d,J＝5.0Hz,4H),6.32(s,2H),6.02(s,2H),5.94(s,2H),5.02(s,2H),3.83(s,6H),3.54(s,1H),0.76(s,4H),0.22(s,4H).
compound MX2 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ182.26(s),164.58(s),162.48(s),160.69(s),159.06(s),154.54(s),148.51(s),124.08(s),122.29(s),115.10(s),113.44(s),104.41(s),103.45(s),100.08(s),95.00(s),58.77(s),56.83(s),7.98(s).
2. by using the method, compounds MX1, MX4, HHQ2, YD2, SFH, YD3, HHQ3 and YM2 can be synthesized respectively, which are as follows:
compound MX1 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ7.62(s,1H),7.13(d,J＝5.0Hz,2H),6.41(s,1H),6.02(s,1H),5.94(s,1H),5.03(s,1H),3.84(d,J＝10.0Hz,6H).
compound MX1 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ182.26(s),164.58(s),162.48(s),160.69(s),159.06(s),152.40(s),150.69(s),122.87(s),121.14(s),112.92(s),110.92(s),104.41(s),103.45(s),100.08(s),95.00(s),56.83(s).
compound MX4 hydrogen profile: 1 H NMR(400MHz,DMSO-d 6 )δ7.61(d,J＝8.5Hz,9H),7.13(d,J＝5.0Hz,12H),6.36(s,6H),6.02(s,6H),5.94(s,6H),5.03(s,6H),3.49(s,5H),0.67(s,12H),0.41(s,12H).
compound MX4 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ182.26(s),164.58(s),162.48(s),160.69(s),159.06(s),153.28(s),149.39(s),125.50(s),124.01(s),122.50(s),118.56(s),116.59(s),104.41(s),103.45(s),100.08(s),95.00(s),58.77(s),7.98(s).
compound HHQ2 hydrogen profile: 1 H NMR(400MHz,DMSO-d 6 )δ7.62(s,1H),7.13(d,J＝5.0Hz,2H),6.45(s,1H),6.02(s,1H),4.90(s,1H),3.84(d,J＝10.0Hz,6H),3.71(s,3H).
compound HHQ2 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ182.41(s),163.23(s),157.49(s),156.43(s),152.75(s),152.40(s),150.69(s),125.62(s),122.87(s),121.14(s),112.92(s),110.92(s),104.68(s),100.84(s),98.73(s),60.70(s),56.83(s).
compound YD2 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ7.62(s,1H),7.13(d,J＝5.0Hz,2H),6.40(s,1H),6.22(s,1H),3.84(d,J＝10.0Hz,6H),3.71(s,6H).
compound YD2 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ182.23(s),163.23(s),152.40(s),150.69(s),150.48(s),148.76(s),146.79(s),122.93(d,J＝14.9Hz),121.08(d,J＝13.8Hz),112.92(s),110.92(s),104.68(s),103.25(s),60.70(s),56.83(s).
compound YD3 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ8.03(s,3H),7.62(s,6H),7.13(d,J＝5.0Hz,12H),6.41(s,6H),6.25(s,6H),4.67(s,2H),3.71(s,35H),1.32(s,36H).
compound YD3 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ182.23(s),163.23(s),154.51(s),150.48(s),149.77(s),148.76(s),146.79(s),125.65(s),124.97(s),122.92(d,J＝16.6Hz),121.03(s),118.56(s),115.45(s),104.68(s),103.25(s),73.89(s),60.70(s),21.81(s).
compound SFH hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ6.81(s,2H),6.74(s,1H),6.41(s,1H),6.17(s,1H),3.83(s,6H),3.79(s,3H),3.71(s,3H).
compound SFH carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ182.26(s),166.80(s),161.52(s),161.27(s),158.76(s),155.25(s),142.05(s),126.77(s),104.94(d,J＝16.4Hz),103.73(s),98.07(s),93.62(s),60.70(s),56.83(s),56.08(s).
compound HHQ3 hydrogen profile: 1 H NMR(400MHz,DMSO-d 6 )δ8.10(s,1H),7.62(s,2H),7.13(d,J＝5.0Hz,4H),6.37(s,2H),6.02(s,2H),4.88(s,2H),4.13(s,4H),3.71(s,6H),1.42(s,3H).
compound HHQ3 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ182.41(s),163.23(s),157.49(s),156.43(s),156.26(s),152.75(s),149.30(s),126.05(s),125.60(d,J＝4.3Hz),122.33(s),118.56(s),118.30(s),104.68(s),100.84(s),98.73(s),64.46(s),60.70(s),13.80(s).
compound YM2 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ6.79(s,2H),6.43(s,1H),6.01(s,1H),5.93(s,1H),5.02(s,1H),3.82(s,6H),3.70(s,3H).
compound YM2 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ182.26(s),164.58(s),161.52(s),160.69(s),159.06(s),155.25(s),142.05(s),126.77(s),104.87(s),103.73(s),103.45(s),100.08(s),95.00(s),60.70(s),56.83(s).
compound SMX1 hydrogen spectrum: δ 7.62 (s, 3H), 7.13 (d, J =5.0hz, 6H), 6.41 (s, 3H), 6.02 (s, 3H), 5.94 (s, 3H), 5.04 (s, 3H), 4.13 (s, 6H), 3.83 (s, 9H), 1.42 (s, 5H).
Compound SMX1 carbon spectrum: δ 182.26(s), 164.58(s), 162.48(s), 160.69(s), 159.06(s), 155.04(s), 149.94(s), 123.14(s), 121.69(s), 114.86(s), 114.00(s), 104.41(s), 103.45(s), 100.08(s), 95.00(s), 64.46(s), 56.83(s), 13.80(s).
Compound SHQ2 hydrogen spectrum: δ 7.62 (s, 3H), 7.13 (d, J =5.0hz, 6H), 6.39 (s, 3H), 6.02 (s, 3H), 4.89 (s, 3H), 4.13 (s, 6H), 3.83 (s, 9H), 3.71 (s, 9H), 1.42 (s, 5H).
Carbon spectrum of compound SHQ 2: δ 182.41(s), 163.23(s), 157.49(s), 156.43(s), 155.04(s), 152.75(s), 149.94(s), 125.62(s), 123.14(s), 121.69(s), 114.86(s), 114.00(s), 104.68(s), 100.84(s), 98.73(s), 64.46(s), 60.70(s), 56.83(s), 13.80(s).
3. R in the general structure of the synthesis 1 ＝
R 2 ＝-CH 3 、R 3 ＝-H、R 4 ＝-H、R 5 ＝-H、R 6 ＝-H、R 7 ＝-H、R 8 A representative compound of = -OH 2- (3-cyclopentyloxy-4-methoxybenzene) -3,5, 7-trihydroxy-4H-chromen-4-one.
Taking 1.9mmol of 2, 4-dimethoxymethyl ether hydroxyl-protected 6-hydroxy-acetophenone, 3-cyclopentyloxy-4-methoxybenzaldehyde (2.05 mmol), meOH (3.5 mL) and H at room temperature 2 Adding O (9.2 mL) into a reaction bottle, adding pyrrolidine (1.7 mL), raising the temperature to 50 ℃ after the addition is finished, carrying out open stirring reaction for 15h, reacting until TLC monitors that the raw materials disappear, cooling the reaction liquid to room temperature, pouring into ice water, and quenching the reaction. Adjusting pH =4.0 with 1N hydrochloric acid, extracting three times with 12mL dichloromethane, combining the organic phases, washing with saturated sodium chloride solution, drying with anhydrous sodium sulfate, concentrating the organic phase under reduced pressure to dryness, removing the protecting groups in methanol and hydrochloric acid (3 mol/L) under argon protection at room temperature, cooling to room temperature, and performing silica gel column chromatography [ V (petroleum ether): v (acetone) =5]Purifying to obtain the representative compound 2- (3-cyclopentyloxy-4-methoxybenzene) -3,5, 7-trihydroxy-4H-chromen-4-one (HP 2). The data for the hydrogen and carbon spectra of nmr are as follows:
hydrogen spectrum of compound HP 2: 1 H NMR(400MHz,DMSO-d 6 )δ7.61(s,2H),7.13(d,J＝5.0Hz,4H),6.02(s,2H),5.94(s,2H),5.51(s,2H),5.03(s,2H),3.83(s,6H),3.54(s,1H),0.76(s,4H),0.25(s,4H).
compound HP2 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ175.69(s),165.11(s),159.84(s),156.95(s),153.55(s),147.76(d,J＝3.6Hz),137.31(s),124.31(s),124.01(s),113.72(s),113.53(s),103.01(s),99.56(s),95.07(s),58.77(s),56.83(s),7.98(s).
4. using the above method, compounds HP3, HP4, SHT2, SHT3, SCJ2, SCJ3 can be synthesized separately as follows:
compound HP3 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ7.62(s,1H),7.13(d,J＝5.0Hz,2H),6.02(s,1H),5.94(s,1H),5.56(s,1H),5.03(s,1H),3.84(d,J＝10.0Hz,6H).
compound HP3 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ175.69(s),165.11(s),159.84(s),156.95(s),151.56(s),149.79(s),147.77(s),137.31(s),124.09(s),123.25(s),111.87(d,J＝7.0Hz),103.01(s),99.56(s),95.07(s),56.83(s).
compound HP4 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ7.60(d,J＝3.0Hz,3H),7.12(d,J＝5.0Hz,4H),6.01(s,2H),5.91(d,J＝16.5Hz,4H),5.04(s,2H),3.50(s,1H),0.73(s,4H),0.29(s,4H).
compound HP4 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ175.69(s),165.11(s),159.84(s),156.95(s),152.74(s),147.76(d,J＝3.6Hz),137.31(s),126.62(s),125.15(s),122.94(s),118.56(s),114.70(s),103.01(s),99.56(s),95.07(s),58.77(s),7.98(s).
compound SHT2 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ7.62(s,1H),7.13(d,J＝5.0Hz,2H),6.15(d,J＝15.0Hz,2H),5.06(s,1H),3.92(s,3H),3.90–3.72(m,9H).
carbon spectrum of compound SHT 2: 13 C NMR(101MHz,DMSO-d 6 )δ173.15(s),164.47(s),159.57(s),157.80(s),154.41(s),151.82(s),149.93(s),138.59(s),123.83(s),123.08(s),112.85(s),111.93(s),107.78(s),97.52(s),96.77(s),61.51(s),56.83(s).
compound SHT3 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ8.80(s,1H),7.62(s,2H),7.13(d,J＝5.0Hz,4H),6.15(d,J＝15.0Hz,4H),5.06(s,2H),3.92(s,6H),3.84(s,6H),3.56(s,1H),0.74(s,4H),0.31(s,4H).
compound SHT3 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )173.15(s),164.47(s),159.57(s),157.80(s),154.41(s),152.59(s),148.63(s),138.59(s),125.96(s),124.85(s),123.94(s),118.56(s),115.17(s),107.78(s),97.52(s),96.77(s),61.51(s),58.77(s),56.83(s),7.98(s).
compound SHP3 hydrogen spectrum: δ 7.62 (s, 3H), 7.13 (d, J =5.0hz, 6H), 6.02 (s, 3H), 5.94 (s, 3H), 5.53 (s, 3H), 5.03 (s, 3H), 4.13 (s, 6H), 3.83 (s, 9H), 1.42 (s, 5H).
Compound SHP3 carbon spectrum: δ 175.69(s), 165.11(s), 159.84(s), 156.95(s), 154.39(s), 148.68(s), 147.77(s), 137.31(s), 124.04(s), 123.57(s), 114.46(s), 113.75(s), 103.01(s), 99.56(s), 95.07(s), 64.46(s), 56.83(s), 13.80(s).
Compound SHT5 hydrogen spectrum: δ 7.62 (s, 1H), 7.13 (d, J =5.0hz, 2h), 6.15 (d, J =15.0hz, 2h), 5.07 (s, 1H), 4.13 (s, 2H), 3.92 (s, 3H), 3.84 (d, J =5.0hz, 6h), 1.42 (s, 3H).
Compound SHT5 carbon spectrum: δ 173.15(s), 164.47(s), 159.57(s), 157.80(s), 154.41(s), 151.55(s), 149.13(s), 138.59(s), 124.99(s), 123.38(s), 115.90(s), 113.46(s), 107.78(s), 97.52(s), 96.77(s), 64.46(s), 61.51(s), 56.83(s), 13.80(s).
Compound SCJ2 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ7.62(s,1H),7.13(d,J＝5.0Hz,2H),6.02(s,1H),5.01(s,1H),3.87–3.81(m,9H),3.71(s,3H).
compound SCJ2 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ179.30(s),158.57(s),154.43(d,J＝6.0Hz),153.75(s),151.82(s),149.93(s),138.59(s),128.86(s),123.83(s),123.08(s),112.85(s),111.93(s),104.77(s),95.28(s),61.51(s),60.70(s),56.83(s).
compound SCJ3 hydrogen spectrum: 1 H NMR(400MHz,DMSO-d 6 )δ7.73(s,3H),7.62(s,8H),7.13(d,J＝5.0Hz,12H),6.02(s,6H),4.89(s,6H),3.84(s,18H),3.71(s,18H),3.49(s,5H),0.73(s,12H),0.24(s,12H).
compound SCJ3 carbon spectrum: 13 C NMR(101MHz,DMSO-d 6 )δ179.30(s),158.57(s),154.43(d,J＝6.0Hz),153.75(s),152.59(s),148.63(s),138.59(s),128.86(s),125.96(s),124.85(s),123.94(s),118.56(s),115.17(s),104.77(s),95.28(s),61.51(s),60.70(s),58.77(s),7.98(s).
example 2 novel Chinese medicinal flavone derivatives inhibit Phosphodiesterase (PDEs)
PDE4 activity was determined in enzyme preparations from rat polymorphonuclear lymphocytes (PMNL) and PDE2, PDE3 and PDE5 activity was determined using isolated platelet PDE. Preventing coagulation of the extracted rat blood with citrate; separating the platelet rich plasma from the red and white blood cells in the supernatant by centrifugation at room temperature; platelets were then lysed by sonication and reserved for use in the PDE3 and PDE5 assays. To determine PDE2 activity, the cytoplasmic platelet fraction was purified by NaCl gradient on an anion exchange column, obtaining the PDE2 peak for the determination; after further sedimentation by dextran, PMNL cells for PDE4 assay were isolated by Ficoll gradient centrifugation. After washing the PMNL cells 2 times, residual erythrocytes were removed by lysis for 6 minutes at 4 ℃ by adding 10mL hypotonic lysis buffer. The still intact PMNL cells were washed twice more with PBS and by sonication, centrifuged at high speed for 1h at 4 ℃ and the supernatant obtained contains the PDE4 cytoplasmic fraction for use as starting material for the enzymes in the PDE4, PDE2, PDE3 and PDE5 assays described below.
The Activity of various PDE enzymes was determined using the phosphodiesterase Activity Assay kit (available from Abcam, inc., cat # ab13940, PDE Activity Assay kit, colorimetric): the operation method is modified appropriately, the protease provided by the kit is replaced by the protease prepared by the method, and other methods are strictly detected according to the operation instruction: adding 20 microliter of cAMP substrate, adding 15 microliter of determination buffer solution, adding 10 microliter of 5' nucleotidase, mixing uniformly, adding test compound with proper concentration, adding extracted and purified PDE enzyme, incubating at 30 ℃ for 30 minutes, adding Green Assay reagent to detect decomposed phosphate ions, mixing for 20 minutes to achieve uniform color, and determining OD620nm to calculate the inhibitory activity of the compound on the PDE enzyme.
Results are shown in Table 1, IC for PDE4 inhibition determined for each compound prepared in example 1 50 Value of 10 -11 -10 -4 M, the selectivity of 2, 3 and 5 types of PDE is a factor of 20-800, which shows that the novel traditional Chinese medicine flavone derivative has the function of inhibiting the activity of PDE4 and can be used as a strong PDE4 inhibitor.
Table 1: IC of traditional Chinese medicine flavone derivative for inhibiting PDE4 50 Value of
| Compound (I) | IC for PDE4 inhibition 50 (μmol/L) |
| MX1 | 0.00071 |
| MX2 | 0.00041 |
| MX4 | 0.00002 |
| HHQ2 | 0.00017 |
| YD2 | 0.00056 |
| SFH | 0.025 |
| YD3 | 0.00017 |
| HHQ3 | 0.00018 |
| YM2 | 0.00051 |
| HP2 | 0.00011 |
| HP3 | 0.00019 |
| HP4 | 0.00007 |
| SHT2 | 0.0012 |
| SHT3 | 0.000065 |
| SCJ2 | 0.00035 |
| SCJ3 | 0.000082 |
| SMX1 | 0.0012 |
| SHQ2 | 0.0073 |
| SHP3 | 0.0056 |
| SHT5 | 0.0062 |
Example 3 novel traditional Chinese medicine flavone derivatives inhibit TNF-alpha release from nasal polyp cells
Nasal polyp tissues stored in the laboratory were washed with RPMI 1640, then lysed with protease (2.5 mg/mL), collagenase (1.0 mg/mL), hyaluronidase (0.5 mg/mL) and DNase (0.1 mg/mL) for 150 minutes at 37 ℃ (1 g of tissue and 4mL of RPMI 1640 containing the enzyme). The resulting mixture of cells (epithelial cells, monocytes, macrophages, lymphocytes, fibroblasts and granulocytes) was filtered, washed by repeated centrifugation in culture solution, passively sensitized by the addition of human IgE, and the cell suspension diluted to a concentration of 2 million cells/mL in RPMI 1640 (supplemented with antibiotics, 10% fetal bovine serum, 2mM glutamine and 25mM Hepes). The suspension was dispensed onto 6 well cell culture plates (1 ml/well). Cells were preincubated for 30 minutes with varying concentrations of test compound and then stimulated to release TNF- α by addition of anti-IgE, with maximum release into the medium occurring after about 16 hours. During this period, the cells were cultured in an incubator containing 5% carbon dioxide at 37 ℃. The medium (supernatant) was collected by centrifugation and kept at-80 ℃ until cytokine determination. TNF- α was measured in the supernatant using an ELISA kit.
Cells not stimulated with anti-IgE produce hardly any TNF- α, whereas stimulated cells secrete large amounts of TNF- α, the amount of TNF- α can be reduced by PDE4 inhibitors, and the degree of reduction is dose-dependent. IC was calculated from the percent inhibition of different concentrations of compound (TNF-alpha release of cells stimulated with anti-IgE = 100%) 50 (concentration to 50% inhibition). Results IC of the compound prepared in example 1 are shown in Table 2 50 Value of 10 -10 -10 -3 M, showing that the traditional Chinese medicine flavone derivative has the function of inhibiting TNF-alpha release, and further showing that the novel traditional Chinese medicine flavone derivative can be used as a TNF-alpha inhibitor.
Table 2: results of inhibition of TNF-alpha Release from Compounds
Example 4 novel Chinese medicinal flavone derivatives inhibit eosinophilia in rat allergy model
50mg of V-grade ovalbumin, 100mg of aluminum hydroxide and inactivated bordetella pertussis 6X 10 9 Dissolving the mixture in 1ml of normal saline to prepare suspension gel, and expanding the suspension gel according to the proportion to prepare 12ml of sensitization liquid for later use (2 ml is prepared for preventing wall-hanging residual loss in 10 injections). Sensitization on day 1: the rats in the blank group are subjected to intraperitoneal injection of 1ml of normal saline, and the rats in the model group are subjected to intraperitoneal injection of 1ml of sensitizing solution for sensitization; challenge started on day 15: placing two groups of rats in atomization boxes with the same size, respectively, and administering physiological salt to blank groupWater 6ml atomization challenge, model group administration 5% v-class ovalbumin solution 6ml atomization challenge, once a day, 30min each challenge, continuous challenge for 10 days. The test compounds were administered intraperitoneally or orally as a suspension in 10% polyethylene glycol 300 and 0.5% 5-hydroxyethyl cellulose 2 hours prior to allergen challenge. The control group was treated with vehicle depending on the administration form of the test compound. Fixing four limbs of a rat at the end of an experiment, disinfecting the neck by using 75% alcohol, fully exposing the trachea, inserting a trachea cannula needle (the needle head is slightly ground flat) near the throat, and inserting the needle head into a certain position without exceeding the bifurcation of the trachea; lavage with 2mL precooled PBS was repeated 3 times, alveolar lavage fluid was collected into EP tubes, centrifuged at 1000rpm at 4 ℃ and cells were collected, stained with Reishi Giemsa and counted under microscope for differential cell counting.
The white blood cells are a vital cell type in the immune process, and the differential cell count can effectively analyze the change of the white blood cell proportion in the alveolar lavage fluid BALF. In the course of the disease, the inflammatory cells infiltrating the bronchi of rats are mainly lymphocytes and eosinophils. Lymphocytes amplify the inflammatory response of eosinophils on the bronchial mucosa and, as eosinophils increase, increase their accumulation, activation and interaction with other inflammatory cells, inflammatory mediators, cytokines in the lung, thereby exacerbating the allergy.
As shown in Table 3, the compound prepared in example 1 effectively inhibited eosinophilia symptoms by 62% -88% after intraperitoneal administration at a dose of 1-10 mg/kg; each compound inhibited eosinophilia by 41-77% after oral administration at a dose of 10-100 mg/kg. Therefore, the novel traditional Chinese medicine flavone derivatives are suitable for preparing medicines for treating and preventing diseases related to eosinophilic activity.
Table 3: results of inhibition of eosinophilia with Compounds
Example 5 novel Chinese medicinal flavone derivatives inhibit Lipopolysaccharide (LPS) -induced neutrophilia
The inhibitory effect of the compound prepared in example 1 on lung neutrophil infiltration was tested in male Wistar rats (200 ± 20 g). On the day of the experiment, animals were individually placed in an open 1L plexiglass box that was connected to a head-nose exposure device. Animals were exposed to an aerosol of lipopolysaccharide suspension (LPS 100 μ g/mL dissolved in PBS solution containing 0.1% hydroxylamine) (LPS challenge) for 45 minutes, and standard controls were sprayed with an aerosol of PBS solution containing 0.1% hydroxylamine for 45 minutes. After 6 hours of LPS challenge, a number of neutrophils migrated into the lungs of the animals. Each compound tested was administered orally as a suspension in 10% polyethylene glycol 300 and 0.5% 5-hydroxyethyl cellulose 2 hours prior to LPS challenge. The control group was treated with the vehicle depending on the administration form of the test substance. Fixing four limbs of a rat at the end of an experiment, disinfecting the neck by using 75% alcohol, fully exposing the trachea, inserting a trachea cannula needle (the needle head is slightly ground flat) near the throat, and inserting the needle head into a certain position without exceeding the bifurcation of the trachea; lavage with 2mL precooled PBS was repeated 3 times, alveolar lavage fluid was collected into EP tubes, centrifuged at 1000rpm at 4 ℃ and cells were collected, stained with Reishi Giemsa and counted under microscope for differential cell counting.
The results are shown in Table 4, and the compounds of the present invention inhibited neutrophilia symptoms by 36% -88% after oral administration at a dose of 10-100 mg/kg. Therefore, the novel traditional Chinese medicine flavone derivative is suitable for preparing medicines for treating and preventing diseases related to the activity of the neutral white blood cells.
Table 4: results of inhibition of neutrophilia by Compounds
Example 6 protective action of novel Chinese medicinal flavone derivatives on A beta aggregate-induced nerve cell injury
The inhibition effect of the compound prepared in example 1 on the nerve cell cytotoxicity induced by the A beta is observed by taking the cell viability without adding the A beta 1-42 as a negative control, and the specific implementation steps are as follows: inoculating PC12 cells into MEM complete culture solution, culturing in 96-well plate, incubating in constant temperature cell incubator for 24 hr, adding the A beta protein oligomer, adding the compound solution to each well after 2 hr, adding equal amount of sterile water to the model group, and incubating for 24 hr. After completion, the cell viability was measured by the MTT method. Each time three replicates were performed and the experiment was repeated three times.
The results are shown in table 5, and after treatment with the administration concentration of 50 μmol/L, the cell survival rate is significantly improved compared with that of the model group, which indicates that the compounds of the present invention have a good effect of protecting nerve cells, and have a significant repairing effect on the nerve cell injury induced by the a β aggregates. Therefore, the novel traditional Chinese medicine flavone derivatives are suitable for preparing medicines for treating and preventing diseases related to neuroprotection.
Table 5: results of protecting neuronal cells with Compounds
Example 7 novel Chinese medicinal flavone derivative vs CoCl 2 Protective action for inducing hypoxia injury of nerve cell and cardiac muscle cell
With no addition of CoCl 2 Cell viability of induced nerve cell PC12 and myocardial cell H9C2 was negative control, and the compound prepared in example 1 was observed against induced CoCl 2 The specific implementation steps of the generated inhibition effect of the hypoxia injury of the nerve cells and the cardiac muscle cells are as follows: inoculating PC12 and H9C2 cells into MEM or DMEM complete culture solution, culturing in 96-well plate, incubating in constant temperature cell incubator for 24 hr, adding pre-dissolved CoCl-containing solution 2 After 2h, each compound solution was added to each well of each compound group at a concentration of100 μmol/L, the model group was added with an equal amount of sterile water and incubation continued in the incubator for 48 hours. After completion, the cell viability was measured by the MTT method. Each time three replicates were performed and the experiment was repeated three times.
The results are shown in tables 6 and 7, and after treatment at the administration concentration of 100 μmol/l, the cell survival rates of both nerve cells and myocardial cells are remarkably improved compared with that of a model group, which indicates that the compounds of the invention have better effects of protecting the nerve cells and the myocardial cells from hypoxia injury. Therefore, the novel traditional Chinese medicine flavone derivative is suitable for preparing medicines for treating and preventing diseases related to the damage of nerve cells and cardiovascular systems.
Table 6: effect of Compounds on hypoxia-induced nerve Damage
| Group of | Cell survival rate (%) |
| Model set | 53±2.8 |
| MX1 | 60±1.2 |
| MX2 | 71±2.0 |
| MX4 | 88±2.7 |
| HHQ2 | 92±3.7 |
| YD2 | 82±3.2 |
| SFH | 60±2.1 |
| YD3 | 81±2.2 |
| HHQ3 | 77±2.1 |
| YM2 | 78±2.0 |
| HP2 | 73±1.5 |
| HP3 | 85±2.1 |
| HP4 | 95±3.7 |
| SHT2 | 87±3.1 |
| SHT3 | 92±3.6 |
| SCJ2 | 78±2.1 |
| SCJ3 | 90±2.7 |
| SMX1 | 81±2.6 |
| SHQ2 | 88±3.7 |
| SHP3 | 82±3.3 |
| SHT5 | 72±2.8 |
Table 7: protective effect of compound on cardiovascular cells caused by hypoxia
Example 8 inhibition of oxidative Low-Density lipoprotein (ox-LDL) -induced vascular endothelial cell injury by novel traditional Chinese medicine flavone derivatives
HUVEC of vascular endothelial cells are inoculated in MEM complete culture solution, and the protective effect of the compound on vascular endothelial atherosclerosis injury induced by ox-LDL is observed by taking a group of non-oxidized low-density lipoprotein ox-LDL as a negative control. The method comprises the following specific steps: HUVEC cells are inoculated in MEM or DMEM complete culture solution, placed into a 96-well plate for culture, placed into a constant-temperature cell incubator for incubation for 24 hours, then induction liquid containing ox-LDL dissolved in advance is added, and after 2 hours, 50 mu M of traditional Chinese medicine flavone derivative is added into each well, and a group without adding ox-LDL is used as a negative control. After finishing, the content of IL-6 inflammatory factors is detected by ELISA.
The results are shown in table 8, compared with the model group, the novel traditional Chinese medicine flavone derivative can effectively inhibit the increase of IL-6 in the vascular endothelial cells stimulated by oxidative low-density lipoprotein ox-LDL, and the novel traditional Chinese medicine flavone derivative is suitable for preparing the medicines for treating the diseases related to the injury of the vascular endothelial cells.
Table 8: inhibition of ox-LDL induced endothelial cell atheroma
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described in the foregoing embodiments, or equivalents may be substituted for some of the features thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Claims (10)
1. A novel traditional Chinese medicine flavone derivative is characterized in that the novel traditional Chinese medicine flavone derivative has a structure of the following general formula:
wherein R is 1 Selected from fluorine-containing alkyl group consisting of 1-5 carbon atoms, cycloalkyl or cycloalkylmethyl group consisting of 3-6 carbon atoms, or saturated or unsaturated alkyl group of C2-C6; r is 2 Selected from hydrogen or fluorine-containing alkyl group consisting of 1-5 carbon atoms, cycloalkyl or cycloalkylmethyl group consisting of 3-6 carbon atoms, or saturated or unsaturated alkyl group of C2-C6; r 3 Selected from hydrogen or alkoxy; r is 4 Selected from hydrogen or alkyl; r 5 Selected from hydrogen or alkyl; r 6 Selected from hydrogen, hydroxy or alkoxy; r is 7 Selected from hydrogen or alkoxy; r is 8 Selected from hydrogen, hydroxy or alkoxy.
2. The novel traditional Chinese medicine flavone derivative of claim 1, wherein the novel traditional Chinese medicine flavone derivative has a structure of the following general formula:
wherein R is 1 Is selected from-CH 3 、
Or
R 2 Is selected from-CH 3 、
Or alternatively
R 3 Is selected from-H or-OCH 3 ；R 4 Is selected from-H or-CH 3 ；R 5 Is selected from-H or-CH 3 ；R 6 Is selected from-H or-OH or-OCH 3 ；R 7 Is selected from-H or-OCH 3 ；R 8 Is selected from-H, -OH or-OCH 3 。
3. The novel chinese medicinal flavone derivative according to claim 1, wherein the novel chinese medicinal flavone derivative is MX1, MX2, MX4, HHQ2, YD2, SFH, YD3, HHQ3, YM2, HP3, HP4, SHT2, SHT3, SCJ2, SCJ3, SMX1, SHQ2, SHP3, SHT5, which specifically comprises the following structure:
4. the method for preparing a novel flavone derivative of a traditional Chinese medicine according to claim 3, comprising the steps of:
mixing a derivative X of 2-methoxyacetophenone protected by MOM hydroxyl with a derivative Y of benzaldehyde, carrying out nucleophilic addition under the catalysis of alkali, generating chalcone by using a claisen-Schmidt method, carrying out intramolecular cyclization, and removing a protecting group under an acidic condition to obtain novel traditional Chinese medicine flavone derivatives MX1, MX2, MX4, HHQ2, YD2, SFH, YD3, SMX1, SHQ2, HHQ3 and YM2;
or mixing MOM hydroxyl protected 2-hydroxyacetophenone derivative Z with benzaldehyde derivative Y, adding pyrrolidine to react completely, removing protecting groups, extracting, washing, drying, concentrating, and purifying to obtain new traditional Chinese medicine flavone derivatives HP2, HP3, HP4, SHT2, SHT3, SCJ2, SHP3, SHT5 and SCJ3;
the structure of the derivative X is as follows:
the structure of the derivative Y is as follows:
the structure of the derivative Z is as follows:
wherein R is 1 Is selected from-CH 3 、
Or alternatively
R 2 Is selected from-CH 3 、
Or alternatively
R 3 Is selected from-H or-OCH 3 ；R 4 Is selected from-H or-CH 3 ；R 5 Is selected from-H or-CH 3 ；R 6 Is selected from-H or-OH or-OCH 3 ；R 7 Is selected from-H or-OCH 3 。
5. Use of novel chinese medicinal flavone derivatives according to any one of claims 1 to 3 for the preparation of phosphodiesterase PDEs and/or TNF- α inhibitors.
6. Use of novel chinese medicinal flavone derivatives according to any one of claims 1 to 3 for the preparation of a medicament for the prevention and treatment of inflammatory and/or allergic diseases.
7. The use of the novel Chinese medicinal flavone derivatives according to claim 6 for preparing a medicament for preventing and treating inflammatory and/or allergic diseases, wherein the novel Chinese medicinal flavone derivatives can relieve the symptoms of inflammatory and/or allergic diseases by effectively inhibiting the activity of phosphodiesterase PDEs, inhibiting the release of TNF-alpha, and inhibiting the increase of neutrophils and eosinophils.
8. Use of novel chinese medicinal flavone derivatives according to any one of claims 1 to 3 for the preparation of a medicament for the prevention and treatment of diseases of the nervous system and/or cardiovascular system.
9. The use of the novel derivatives of Chinese medicinal flavonoids according to claim 8 for the preparation of a medicament for the prevention and treatment of diseases of the nervous system and/or cardiovascular system, wherein the novel derivatives of Chinese medicinal flavonoids are capable of inhibiting effectively the a β aggregates and/or CoCl 2 Induced nerve cell and/or myocardial cell damage, or effective reversal of oxidative low density lipoprotein ox-LDL inductionThe vascular endothelial cells are damaged, thereby achieving the effect of protecting nerve cells or cardiovascular cells.
10. The application of the novel traditional Chinese medicine flavone derivative disclosed by claim 6 in preparing a medicine for preventing and treating inflammatory and/or allergic diseases, or the application of the novel traditional Chinese medicine flavone derivative disclosed by claim 8 in preparing a medicine for preventing and treating diseases of a nervous system and/or a cardiovascular system, is characterized in that the medicine is tablets, oral liquid, aerosols, pills, capsules, granules, paste, dripping pills, syrup, powder, granules, tinctures, powder injections or injection solutions.
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| CN115197190A (en) * | 2021-04-09 | 2022-10-18 | 中国海洋大学 | Novel plant flavone derivative and preparation method and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115197190A (en) * | 2021-04-09 | 2022-10-18 | 中国海洋大学 | Novel plant flavone derivative and preparation method and application thereof |
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