CN113149857B - Amide compound and preparation method and application thereof - Google Patents
Amide compound and preparation method and application thereof Download PDFInfo
- Publication number
- CN113149857B CN113149857B CN202110504151.0A CN202110504151A CN113149857B CN 113149857 B CN113149857 B CN 113149857B CN 202110504151 A CN202110504151 A CN 202110504151A CN 113149857 B CN113149857 B CN 113149857B
- Authority
- CN
- China
- Prior art keywords
- amide compound
- ethyl acetate
- uncaria
- compound
- compound according
- 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
- -1 Amide compound Chemical class 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 150000001875 compounds Chemical class 0.000 claims abstract description 29
- 241000157373 Uncaria rhynchophylla Species 0.000 claims abstract description 10
- 239000000126 substance Substances 0.000 claims abstract description 6
- 241001107098 Rubiaceae Species 0.000 claims abstract description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 18
- 241000157352 Uncaria Species 0.000 claims description 16
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- 239000002024 ethyl acetate extract Substances 0.000 claims description 14
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 12
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 12
- 239000000287 crude extract Substances 0.000 claims description 11
- 238000001514 detection method Methods 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 239000012153 distilled water Substances 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 6
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 5
- 238000004811 liquid chromatography Methods 0.000 claims description 5
- 239000000741 silica gel Substances 0.000 claims description 5
- 229910002027 silica gel Inorganic materials 0.000 claims description 5
- 238000002791 soaking Methods 0.000 claims description 5
- 239000004480 active ingredient Substances 0.000 claims description 4
- 238000000034 method Methods 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 3
- 230000014759 maintenance of location Effects 0.000 claims description 3
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 238000010298 pulverizing process Methods 0.000 claims description 2
- 150000003839 salts Chemical class 0.000 claims description 2
- 239000000725 suspension Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 3
- 229940121363 anti-inflammatory agent Drugs 0.000 claims 1
- 239000002260 anti-inflammatory agent Substances 0.000 claims 1
- 230000003110 anti-inflammatory effect Effects 0.000 abstract description 6
- 239000003814 drug Substances 0.000 abstract description 5
- 239000002994 raw material Substances 0.000 abstract 1
- 238000005481 NMR spectroscopy Methods 0.000 description 10
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 description 10
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- 239000002609 medium Substances 0.000 description 8
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 7
- 206010061218 Inflammation Diseases 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 7
- 230000004054 inflammatory process Effects 0.000 description 7
- 238000001228 spectrum Methods 0.000 description 7
- 238000002835 absorbance Methods 0.000 description 5
- 229940109262 curcumin Drugs 0.000 description 5
- 235000012754 curcumin Nutrition 0.000 description 5
- 239000004148 curcumin Substances 0.000 description 5
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000012091 fetal bovine serum Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 229920006395 saturated elastomer Polymers 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 108010019160 Pancreatin Proteins 0.000 description 2
- 229930013930 alkaloid Natural products 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 2
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 241000208340 Araliaceae Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 1
- MUVGVMUWMAGNSY-YCBKVTMHSA-N Corynoxeine Natural products O=C(OC)/C(=C\OC)/[C@@H]1[C@@H](C=C)CN2[C@H]([C@@]3(C(=O)Nc4c3cccc4)CC2)C1 MUVGVMUWMAGNSY-YCBKVTMHSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 244000081822 Uncaria gambir Species 0.000 description 1
- 241000123747 Uncaria hirsuta Species 0.000 description 1
- 241000123748 Uncaria macrophylla Species 0.000 description 1
- 229930187904 Uncarine Natural products 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 229940124599 anti-inflammatory drug Drugs 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 239000002021 butanolic extract Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 239000011365 complex material Substances 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000002035 hexane extract Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- MUVGVMUWMAGNSY-KAXDATADSA-N methyl (e)-2-[(3r,6'r,7's,8'as)-6'-ethenyl-2-oxospiro[1h-indole-3,1'-3,5,6,7,8,8a-hexahydro-2h-indolizine]-7'-yl]-3-methoxyprop-2-enoate Chemical compound O=C1NC2=CC=CC=C2[C@]21CCN1[C@H]2C[C@H](\C(=C/OC)C(=O)OC)[C@@H](C=C)C1 MUVGVMUWMAGNSY-KAXDATADSA-N 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 229940002612 prodrug Drugs 0.000 description 1
- 239000000651 prodrug Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000012453 solvate Substances 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
- C07C235/88—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having the nitrogen atom of at least one of the carboxamide groups further acylated
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention belongs to the field of chemical medicine, and particularly relates to an amide compound, and a preparation method and application thereof. In particular to a novel amide compound which is obtained by taking dry stems and branches of uncaria rhynchophylla of rubiaceae as raw materials and separating through certain preparation steps. The invention discloses a specific structure of the compound, and the anti-inflammatory activity of the compound is measured, and the result shows that the compound has obvious anti-inflammatory activity.
Description
Technical Field
The invention belongs to the field of chemical medicine, and particularly relates to an amide compound, and a preparation method and application thereof.
Background
Uncaria rhynchophylla (Uncaria rhynchophylla), also known as Uncaria gambir, Uncaria oblonga, Araliaceae, evergreen vine, etc. Uncaria rhynchophylla is classified into Uncaria macrophylla Wall, Uncaria hirsuta Wall, Uncaria sinica Wall or Uncaria sessilifolia Wall. The proper growing environment of uncaria is warm and moist, loose and fertile soil, and the uncaria is planted in Guangdong, Guizhou, Guangxi, Fujian, Jiangxi and the like. The medicinal part of uncaria is usually the branch with hook, which is sweet and bitter in taste and slightly cold in flavor, enters heart meridian and has the function of clearing heat and expelling pathogenic factors. More than 100 chemical components are reported to be separated from uncaria rhynchophylla at present, and comprise alkaloids, flavonoids, triterpenes, glycosides and the like, wherein the content of the alkaloids is particularly rich, such as uncarine, isocoubine, corynoxeine and the like. The pharmacological actions of uncaria are closely related to the chemical components of uncaria, such as diminishing inflammation, relieving pain, reducing blood pressure, resisting cancer, resisting epilepsy and the like.
The rapid separation and purification technology of traditional Chinese medicine components has important significance for understanding the complex material basis of traditional Chinese medicine, controlling important quality and discovering potential active substances. The high-pressure preparation liquid phase system greatly improves the sample loading amount on the premise of ensuring the separation degree of a sample on the basis of the analytical high performance liquid chromatography, thereby quickly obtaining a high-purity target compound. The high-pressure preparation liquid phase is widely applied to the separation research of samples such as traditional Chinese medicines, biological products, foods and the like.
Disclosure of Invention
The object of the present invention is to provide a novel amide compound. Another object of the present invention is to provide a method for preparing the compound and its use.
The amide compound has the chemical structural formula as follows:
the preparation method of the amide compound comprises the following steps:
(1) pulverizing dried stem and branch of Uncaria rhynchophylla of Rubiaceae, adding ethanol-water (70% -30%), cold soaking, filtering, repeating for 3-5 times, mixing extractive solutions, concentrating under reduced pressure, and drying to obtain crude extract;
(2) dissolving the crude extract in distilled water to obtain suspension, sequentially extracting with n-hexane, ethyl acetate and n-butanol, retaining ethyl acetate extract phase to obtain ethyl acetate extract, concentrating the ethyl acetate extract under reduced pressure, and drying to obtain ethyl acetate extract;
(3) passing the ethyl acetate extract through silica gel column, gradient eluting, and dividing into 9 fractions, which are respectively named as Fr.1-Fr.9; fr.3 is taken to pass through liquid chromatography, and 66 percent methanol-water to obtain the compound shown in the formula I, namely the amide compound.
In the above extraction method, wherein
In the step (1), the mass-volume ratio of the uncaria to 70% ethanol water is 1: 7-13, preferably 1:10, the unit of mass-to-volume ratio is kg/L. For example, the mass-to-volume ratio is 1:10, i.e. 10L of 70% ethanol water is required to be added into 1kg of uncaria; the cold soaking time is 13-17 days, preferably 15 days; the number of repetitions is preferably 3.
In the step (2), the mass-volume ratio of the uncaria crude extract to distilled water is 1: 1-2; the mass volume ratio of the uncaria crude extract to the n-hexane, the ethyl acetate and the n-butanol is 1: 0.5-2, preferably 1: 1-1.2, and the unit of the mass-volume ratio is kg/L. For example, 1L n-hexane, 1L ethyl acetate, 1L n-butanol may be added to 1:1 mass/volume ratio, i.e., 1kg of uncaria crude extract.
In the step (3), a mixed elution system of dichloromethane and methanol is adopted when the ethyl acetate extract passes through a silica gel column, and the volume ratio of the dichloromethane to the methanol is 9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8 and 1:9 in sequence; the elution system was changed every five column volumes.
In step (3), the specification of the chromatographic column in the liquid chromatography is Innocal ODS-2, 10X 250mm,5 μm. The flow rate is 3mL/min, the column temperature is 25 ℃, the detection is dual-wavelength detection, 254nm and 280nm respectively, and the retention time is 22 min.
The invention also provides a pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable carrier; the active ingredient comprises the amide compound or pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, prodrug molecule, metabolite or mixture thereof.
The invention also relates to application of the amide compound or the pharmaceutical composition in preparation of anti-inflammatory drugs. Wherein, the amide compound shows the capability of inhibiting inflammatory cells, and the in vitro anti-inflammatory activity detection is carried out on the amide compound by adopting an LPS induced RAW264.7 inflammation model method, and the result shows that the inflammation inhibition rate is 90.91%.
Drawings
FIG. 1 Pre-HPLC chromatogram of Uncaria rhynchophylla extract
FIG. 2 NMR of Compounds of formula I1H spectrogram
FIG. 3 NMR of Compounds of formula I13C spectrum
FIG. 4 nuclear magnetic resonance DEPT135 spectrum of compound of formula I
FIG. 5 NMR HMBC spectrum of the compound of formula I
FIG. 6 NMR HSQC spectra of compounds of formula I
Detailed Description
The invention is further illustrated by the following examples. The following examples must be interpreted to illustrate the invention without limiting it. Simple modifications of the invention in accordance with its spirit fall within the scope of the claimed invention.
Some of the instruments and reagents used in the following examples were: pujindo TU-1950 ultraviolet spectrophotometer (Beijing Pujindo); model METTLER AE240 electronic balance (mettler-toledo (shanghai) ltd), KQ-250B ultrasonic cleaner (kunshan ultrasonic instruments ltd). CO 22An incubator; 96-well cell culture plates; an AM-100 ultraviolet spectrophotometer; DEME medium, fetal bovine serum (FBS, Gibco); LPS, DMSO (Sigma company); CCK8 kit; a cell counter; the other reagents were analytical grade, and the water was distilled water. Other instruments are common laboratory instruments; the other reagents are also conventional reagents and are commercially available or available in the laboratory.
EXAMPLE 1 extraction of amides of formula I
The amide compound of formula I is prepared by the following method:
(1) weighing 10kg of uncaria rhynchophylla, dividing into 2 batches, each 5kg of uncaria rhynchophylla, adding 50L of ethanol water (70% -30%) into each batch, carrying out cold soaking for 15 days, repeating for 3 times, combining extracting solutions, concentrating under reduced pressure, and drying to obtain 940.3g of crude extract.
(2) Dissolving the crude extract in 1L of distilled water, placing in a 3L separating funnel, adding n-hexane 1L, extracting for 3 times, mixing the extractive solutions, concentrating under reduced pressure, and drying to obtain n-hexane extract 5.1 g. For the remaining aqueous phase, extraction was carried out in the same manner using 1L of ethyl acetate and 1L of n-butanol in this order to obtain 9.4g of ethyl acetate extract and 91.9g of n-butanol extract, respectively, and the aqueous layer was 830 g.
(3) Passing the ethyl acetate extract through silica gel column (dichloromethane: methanol 9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8, 1:9), changing the elution system every five column volumes, and performing gradient elution to divide the mixture into 9 components which are respectively named as Fr.1-Fr.9. Subjecting Fr.3 to liquid chromatography, and subjecting to 66% methanol-water to obtain amide compound of formula I. The liquid chromatography column specification is Innocal ODS-2, 10X 250mm,5 μm. The flow rate is 3mL/min, the column temperature is 25 ℃, the detection is dual-wavelength detection, 254nm and 280nm respectively, and the retention time is 22 min. As shown in fig. 1.
The structure of the obtained compound is identified, and the identification result is as follows:
hydrogen spectrum: 7.00(dd, J ═ 8.4,1.8Hz,1H, H-2),6.71(d, J ═ 8.4Hz,1H, H-3),7.51(d, J ═ 1.8Hz,1H, H-6),6.66(d, J ═ 12.6Hz,1H, H-7),5.86(d, J ═ 12.6Hz,1H, H-8),7.02(dd, J ═ 8.4,1.8Hz,1H, H-2 '), 6.82(d, J ═ 8.4Hz,1H, H-3'), 7.15(d, J ═ 1.8Hz,1H, H-6 '), 7.49(d, J ═ 15.6Hz,1H, H-7'), 6 ', 6.46(d, J ═ 8.81, H, 3H-3, 5H-3, OCH, 5H-6'), and H-63),3.85(s,5’-OCH3)。
Carbon spectrum: 128.5(C-1),125.3(C-2),115.9(C-3),148.8(C-4),148.6(C-5),114.2(C-6),139.2(C-7),120.9(C-8),173.0(C-9),128.2(C-1 '), 123.5(C-2 '), 116.6(C-3 '), 149.5(C-4 '), 150.2(C-5 '), 111.7(C-6 '), 141.7(C-7 '), 118.3(C-8 '), 171.7(C-9 '), 56.54 (5-OCH)3),56.46(5’-OCH3)。
Nuclear magnetic resonance of the compound1The spectrum H is shown in figure 2.
Nuclear magnetic resonance of the compound13The spectrum C is shown in FIG. 3.
The nuclear magnetic resonance DEPT135 spectrum of the compound is shown in figure 4.
The compound has a nuclear magnetic resonance HMBC spectrum as shown in figure 5.
The nuclear magnetic resonance HSQC spectrum of the compound is shown in FIG. 6.
Determining the structural formula as follows:
EXAMPLE 2 study of the anti-inflammatory Activity of Compounds of formula I
Preparing a test article: taking the compound of the formula I, preparing 50mg/mL stock solution by DMSO, and diluting the compound into 50 mu g/mL test sample liquid by serum-free DMEM medium when dosing. In addition, curcumin is taken and prepared into 50mg/mL stock solution by DMSO, and is diluted into 50 mug/mL test sample liquid by serum-free DMEM medium when dosing.
Cell culture: RAW264.7 cells were cultured in DMEM containing 10% FBS and 5% CO2And cultured in a cell culture box at 37 ℃ and saturated humidity. And (4) carrying out passage for 1 time after the cells grow to the logarithmic growth phase and carrying out passage for 2-3 d. Cells after at least 3 passages were taken for experiments.
Effect of compounds on growth of RAW264.7 cells: digesting RAW264.7 cells in logarithmic phase with 0.25% pancreatin-0.02% EDTA for about 2min, discarding pancreatin, neutralizing pancreatin with 10% FBS DMEM medium, gently blowing to obtain single cell suspension, centrifuging, discarding supernatant, resuspending cells with complete medium, counting, and adjusting cell suspension to 3 × 105Perml, 100. mu.L per well was inoculated into 96-well plates at 37 ℃ with 5% CO2Culturing for 24h under saturated humidity condition, completely sucking supernatant from each well, adding 100 μ L serum-free DMEM medium into each well, and dividing into control group, positive group (curcumin), and compound experiment group of formula I; an additional cell-free blank was set (100. mu.L serum-free DMEM medium was added directly to the wells). Each group is provided with 3 multiple wells, and after adding corresponding drugs (wherein, 100 μ L of the compound of the formula I with the concentration of 50 μ g/mL is added into each well of the compound of the formula I experimental group, 100 μ L of curcumin with the concentration of 50 μ g/mL is added into each well of the positive group, and 100 μ L of serum-free DMEM medium is added into each well of the control group and the blank group), 5% CO is added2Culturing at 37 deg.C under saturated humidity for 24 hr. Then, adding CCK-8 according to 10 mu L of each well, detecting the absorbance value at 450nm by an enzyme-linked immunosorbent assay after 4h (A)450). The inflammation inhibition rate of the compound was calculated according to the following formula, wherein ApostiveAbsorbance value for positive group, AcontrolAs absorbance value of control group, AtreatedIs the absorbance value of the test group of the compound of formula I, AblankBlank absorbance values.
Inflammation inhibition rate [1- (A)treated-Ablank)/(Acontrol-Ablank)]×100％
The anti-inflammatory experimental results show that Apostive＝0.099，Atreated＝0.105，Acontrol＝0.185，Ablank0.097. The positive group curcumin has the inflammation inhibition rate of 97.72 percent and the compound shown in the formula I has the inflammation inhibition rate of 90 percent through calculation91%. The anti-inflammatory effect of the compound of formula I is equivalent to that of curcumin.
Claims (7)
1. An amide compound characterized by having a chemical structural formula shown in formula I:
2. a method for producing the amide compound according to claim 1, comprising the steps of:
(1) pulverizing dried stem and branch of Uncaria rhynchophylla of Rubiaceae, adding 70% ethanol-water, cold soaking, filtering, repeating for 3-5 times, mixing extractive solutions, concentrating under reduced pressure, and drying to obtain crude extract;
(2) dissolving the crude extract in distilled water to obtain suspension, sequentially extracting with n-hexane, ethyl acetate and n-butanol, retaining ethyl acetate extract phase to obtain ethyl acetate extract, concentrating the ethyl acetate extract under reduced pressure, and drying to obtain ethyl acetate extract;
(3) passing the ethyl acetate extract through silica gel column, gradient eluting, and dividing into 9 fractions, which are respectively named as Fr.1-Fr.9; fr.3 is taken to pass through liquid chromatography, and 66 percent methanol-water to obtain the compound shown in the formula I, namely the amide compound.
3. The method for producing an amide compound according to claim 2, wherein in step (1), the mass-to-volume ratio of the uncaria stem with respect to 70% ethanol-water is 1: 7-13; the cold soaking time is 13-17 days; the number of repetitions was 3.
4. The method for producing an amide compound according to claim 2, wherein in step (2), the mass-to-volume ratio of the uncaria crude extract to distilled water is 1: 1-2; the mass volume ratio of the uncaria crude extract to the n-hexane, the ethyl acetate and the n-butanol is 1: 0.5-2.
5. The process for producing an amide compound according to claim 2, wherein, in the step (3),
when the ethyl acetate extract passes through a silica gel column, a mixed elution system of dichloromethane and methanol is adopted, wherein the volume ratio of the dichloromethane to the methanol is 9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8 and 1:9 in sequence; the elution system was changed every five column volumes;
the specification of a chromatographic column in the liquid chromatogram is incal ODS-2,10 x 250mm,5 μm; the flow rate is 3mL/min, the column temperature is 25 ℃, the detection is dual-wavelength detection, 254nm and 280nm respectively, and the retention time is 22 min.
6. A pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable carrier; the active ingredient comprising the amide compound according to claim 1 or a pharmaceutically acceptable salt thereof.
7. Use of the amide compound according to claim 1 or the pharmaceutical composition according to claim 6 for the preparation of an anti-inflammatory agent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110504151.0A CN113149857B (en) | 2021-05-10 | 2021-05-10 | Amide compound and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110504151.0A CN113149857B (en) | 2021-05-10 | 2021-05-10 | Amide compound and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113149857A CN113149857A (en) | 2021-07-23 |
| CN113149857B true CN113149857B (en) | 2021-10-29 |
Family
ID=76874106
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110504151.0A Active CN113149857B (en) | 2021-05-10 | 2021-05-10 | Amide compound and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113149857B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6039949A (en) * | 1997-02-27 | 2000-03-21 | Campamed, Inc. | Method of preparation and composition of a water soluble extract of the plant species uncaria |
| CN102309576A (en) * | 2011-07-05 | 2012-01-11 | 南京泽朗农业发展有限公司 | Preparation method of uncaria total alkaloids |
| ES2478690A1 (en) * | 2012-12-18 | 2014-07-22 | Consejo Superior De Investigaciones Científicas (Csic) | Phenolic extracts of uncaria tomentosa l. (cat's claw) containing procyanidins, propelargonidins and flavonolignans, method for the production thereof, and applications of same |
| CN107827940A (en) * | 2017-11-20 | 2018-03-23 | 中国科学院昆明植物研究所 | Yncaria stem with hooks acid amides A and its pharmaceutical composition and application |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102274319B (en) * | 2011-07-22 | 2013-05-15 | 福建农林大学 | Preparation method of rhomotoxine hydrolysis product |
-
2021
- 2021-05-10 CN CN202110504151.0A patent/CN113149857B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6039949A (en) * | 1997-02-27 | 2000-03-21 | Campamed, Inc. | Method of preparation and composition of a water soluble extract of the plant species uncaria |
| CN102309576A (en) * | 2011-07-05 | 2012-01-11 | 南京泽朗农业发展有限公司 | Preparation method of uncaria total alkaloids |
| ES2478690A1 (en) * | 2012-12-18 | 2014-07-22 | Consejo Superior De Investigaciones Científicas (Csic) | Phenolic extracts of uncaria tomentosa l. (cat's claw) containing procyanidins, propelargonidins and flavonolignans, method for the production thereof, and applications of same |
| CN107827940A (en) * | 2017-11-20 | 2018-03-23 | 中国科学院昆明植物研究所 | Yncaria stem with hooks acid amides A and its pharmaceutical composition and application |
Non-Patent Citations (2)
| Title |
|---|
| HPLC-MS法同时测定钩藤药材中四种生物碱含量;池雨锋，等;《山东中医药大学学报》;20170531;第262-265页 * |
| Uncaria tomentosa improves insulin sensitivity and inflammation in experimental NAFLD;Layanne C. C.，等;《SCIENTIFIC REPORTS》;20180720;第1-14页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113149857A (en) | 2021-07-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105663118A (en) | Application of flavonoids compound to preparation of neuroprotection medicament | |
| CN115160337A (en) | 1 alpha-alkyl daphnane diterpenoid compounds, and preparation method and application thereof | |
| CN113149857B (en) | Amide compound and preparation method and application thereof | |
| CN110746421A (en) | Extraction method and application of indole monoterpene compound | |
| CN114249784B (en) | Coumarin derivative compound III, extraction method and application thereof | |
| Krstić et al. | Secoiridoids and xanthones in the shoots and roots of Centaurium pulchellum cultured in vitro | |
| CN112898357B (en) | Diterpene glycoside novel compound in trollius chinensis bunge and separation and purification method and application thereof | |
| CN110204589B (en) | Effective component of feather cockscomb seed, extraction method and application thereof in preparing neuroprotective medicament | |
| CN108191936B (en) | A neoline J and K extracted from semen Lepidii, and its preparation method and application | |
| CN109912393B (en) | Method for extracting compound 2',3, 5-trihydroxy-4-methoxy bibenzyl from stem and leaf of dioscorea opposita and application thereof | |
| CN109053641B (en) | Dineolignan compound, and separation preparation method and application thereof | |
| CN108948040B (en) | Gilmaxane type sesquiterpene compound extracted from herba Centellae and application thereof | |
| CN111606801A (en) | Split-ring labdane diterpenoid compound and separation method and application thereof | |
| CN115368420B (en) | Compound with anti-inflammatory activity and preparation method and application thereof | |
| CN107353315A (en) | A kind of method and its application that the new alkali A compounds of northern Roripa are extracted from lepidii,semen | |
| CN115745745B (en) | Method for extracting compound 1, 8-terpineol from ephedra grass stem and application thereof | |
| CN108129534B (en) | A neoline C and D extracted from semen Lepidii, and its preparation method and application | |
| CN112358515B (en) | Application of naringenin (4-O-methyl) glucoside compound in preparation of anti-inflammatory or lipid-lowering drugs | |
| CN110963918B (en) | Sinapic acid derivative and preparation method and application thereof | |
| CN107459467B (en) | A kind of new amine D of Roripa montana, the new amine E of Roripa montana and the preparation method and application thereof | |
| US20070148268A1 (en) | Chemical compounds and their use as therapeutic agent and means of quality assessment of herbal medicines | |
| CN112300239B (en) | Steroid compound in bamboo leaves, its extraction method and use | |
| CN109438535B (en) | Novel benzofuran methyl ketone type compound with anti-gastric cancer effect and extraction and separation method thereof | |
| CN107446008B (en) | Neoside E, and preparation method and application thereof | |
| CN114105765A (en) | Method for preparing chrysanthemum new patch I and chrysanthemum new patch J from stem and leaf of Huai chrysanthemum and application of method |
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 | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240112 Address after: Room 417, Quality Control R&D Pilot Production Workshop, No. 3333 Dazheng Road, High tech Zone, Jinan City, Shandong Province, 250104 Patentee after: Shandong Gujin Traditional Chinese Medicine Technology Co.,Ltd. Address before: No.16369, Jingshi Road, Jinan, Shandong 250355 Patentee before: SHANDONG University OF TRADITIONAL CHINESE MEDICINE |
|
| TR01 | Transfer of patent right |