CN115304466B - Guaiane sesquiterpene compound separated from lilac daphne root, and preparation method and application thereof - Google Patents
Guaiane sesquiterpene compound separated from lilac daphne root, and preparation method and application thereof Download PDFInfo
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- CN115304466B CN115304466B CN202210821138.2A CN202210821138A CN115304466B CN 115304466 B CN115304466 B CN 115304466B CN 202210821138 A CN202210821138 A CN 202210821138A CN 115304466 B CN115304466 B CN 115304466B
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- 241000934856 Daphne Species 0.000 title claims abstract description 19
- -1 Guaiane sesquiterpene compound Chemical class 0.000 title claims abstract description 11
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 241001104043 Syringa Species 0.000 title claims description 10
- 235000004338 Syringa vulgaris Nutrition 0.000 title claims description 10
- 239000003814 drug Substances 0.000 claims abstract description 9
- 150000000176 guaiane derivatives Chemical class 0.000 claims abstract description 9
- 241001310717 Daphne genkwa Species 0.000 claims abstract description 6
- 208000024827 Alzheimer disease Diseases 0.000 claims abstract description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 114
- 150000001875 compounds Chemical class 0.000 claims description 87
- 238000004458 analytical method Methods 0.000 claims description 17
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 9
- 238000010828 elution Methods 0.000 claims description 7
- 239000000284 extract Substances 0.000 claims description 7
- 229930004725 sesquiterpene Natural products 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 238000004440 column chromatography Methods 0.000 claims description 6
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 6
- 238000010898 silica gel chromatography Methods 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 101100313763 Arabidopsis thaliana TIM22-2 gene Proteins 0.000 claims description 3
- 230000007131 anti Alzheimer effect Effects 0.000 claims description 3
- WGLUMOCWFMKWIL-UHFFFAOYSA-N dichloromethane;methanol Chemical compound OC.ClCCl WGLUMOCWFMKWIL-UHFFFAOYSA-N 0.000 claims description 3
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims description 3
- 210000002196 fr. b Anatomy 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
- 239000003208 petroleum Substances 0.000 claims description 3
- 238000004007 reversed phase HPLC Methods 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 2
- 238000010992 reflux Methods 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 1
- 229940022698 acetylcholinesterase Drugs 0.000 abstract description 6
- 229940079593 drug Drugs 0.000 abstract description 6
- 102000012440 Acetylcholinesterase Human genes 0.000 abstract description 5
- 108010022752 Acetylcholinesterase Proteins 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 5
- 230000005764 inhibitory process Effects 0.000 abstract description 4
- 229910052799 carbon Inorganic materials 0.000 description 59
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 45
- 238000005481 NMR spectroscopy Methods 0.000 description 41
- 238000001228 spectrum Methods 0.000 description 41
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 33
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 22
- 230000002596 correlated effect Effects 0.000 description 15
- 229910052739 hydrogen Inorganic materials 0.000 description 14
- 239000001257 hydrogen Substances 0.000 description 14
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 13
- 239000012634 fragment Substances 0.000 description 13
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 13
- 230000005311 nuclear magnetism Effects 0.000 description 13
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 13
- 239000002904 solvent Substances 0.000 description 12
- VILAVOFMIJHSJA-UHFFFAOYSA-N dicarbon monoxide Chemical group [C]=C=O VILAVOFMIJHSJA-UHFFFAOYSA-N 0.000 description 11
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 11
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 11
- 150000002500 ions Chemical class 0.000 description 11
- SNVLJLYUUXKWOJ-UHFFFAOYSA-N methylidenecarbene Chemical group C=[C] SNVLJLYUUXKWOJ-UHFFFAOYSA-N 0.000 description 8
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 7
- 238000005100 correlation spectroscopy Methods 0.000 description 7
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 7
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 7
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical group [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 6
- 229910002091 carbon monoxide Inorganic materials 0.000 description 6
- 229940125782 compound 2 Drugs 0.000 description 6
- 150000002576 ketones Chemical group 0.000 description 6
- GHYOCDFICYLMRF-UTIIJYGPSA-N (2S,3R)-N-[(2S)-3-(cyclopenten-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-hydroxy-3-(4-methoxyphenyl)-2-[[(2S)-2-[(2-morpholin-4-ylacetyl)amino]propanoyl]amino]propanamide Chemical compound C1(=CCCC1)C[C@@H](C(=O)[C@@]1(OC1)C)NC([C@H]([C@@H](C1=CC=C(C=C1)OC)O)NC([C@H](C)NC(CN1CCOCC1)=O)=O)=O GHYOCDFICYLMRF-UTIIJYGPSA-N 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 238000004364 calculation method Methods 0.000 description 5
- 229940125904 compound 1 Drugs 0.000 description 5
- 229940125797 compound 12 Drugs 0.000 description 5
- UNILWMWFPHPYOR-KXEYIPSPSA-M 1-[6-[2-[3-[3-[3-[2-[2-[3-[[2-[2-[[(2r)-1-[[2-[[(2r)-1-[3-[2-[2-[3-[[2-(2-amino-2-oxoethoxy)acetyl]amino]propoxy]ethoxy]ethoxy]propylamino]-3-hydroxy-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-3-[(2r)-2,3-di(hexadecanoyloxy)propyl]sulfanyl-1-oxopropan-2-yl Chemical compound O=C1C(SCCC(=O)NCCCOCCOCCOCCCNC(=O)COCC(=O)N[C@@H](CSC[C@@H](COC(=O)CCCCCCCCCCCCCCC)OC(=O)CCCCCCCCCCCCCCC)C(=O)NCC(=O)N[C@H](CO)C(=O)NCCCOCCOCCOCCCNC(=O)COCC(N)=O)CC(=O)N1CCNC(=O)CCCCCN\1C2=CC=C(S([O-])(=O)=O)C=C2CC/1=C/C=C/C=C/C1=[N+](CC)C2=CC=C(S([O-])(=O)=O)C=C2C1 UNILWMWFPHPYOR-KXEYIPSPSA-M 0.000 description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 4
- 241000628997 Flos Species 0.000 description 4
- 229940125898 compound 5 Drugs 0.000 description 4
- 239000013078 crystal Substances 0.000 description 4
- ADEBPBSSDYVVLD-UHFFFAOYSA-N donepezil Chemical compound O=C1C=2C=C(OC)C(OC)=CC=2CC1CC(CC1)CCN1CC1=CC=CC=C1 ADEBPBSSDYVVLD-UHFFFAOYSA-N 0.000 description 4
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 4
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 150000004354 sesquiterpene derivatives Chemical class 0.000 description 4
- QFLWZFQWSBQYPS-AWRAUJHKSA-N (3S)-3-[[(2S)-2-[[(2S)-2-[5-[(3aS,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]-3-methylbutanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-4-[1-bis(4-chlorophenoxy)phosphorylbutylamino]-4-oxobutanoic acid Chemical compound CCCC(NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)[C@@H](NC(=O)CCCCC1SC[C@@H]2NC(=O)N[C@H]12)C(C)C)P(=O)(Oc1ccc(Cl)cc1)Oc1ccc(Cl)cc1 QFLWZFQWSBQYPS-AWRAUJHKSA-N 0.000 description 3
- 238000004057 DFT-B3LYP calculation Methods 0.000 description 3
- 229940125773 compound 10 Drugs 0.000 description 3
- 238000002212 electronic circular dichroism spectrum Methods 0.000 description 3
- 238000005570 heteronuclear single quantum coherence Methods 0.000 description 3
- ZLVXBBHTMQJRSX-VMGNSXQWSA-N jdtic Chemical compound C1([C@]2(C)CCN(C[C@@H]2C)C[C@H](C(C)C)NC(=O)[C@@H]2NCC3=CC(O)=CC=C3C2)=CC=CC(O)=C1 ZLVXBBHTMQJRSX-VMGNSXQWSA-N 0.000 description 3
- 230000004770 neurodegeneration Effects 0.000 description 3
- 208000015122 neurodegenerative disease Diseases 0.000 description 3
- 238000006467 substitution reaction Methods 0.000 description 3
- VTNULXUEOJMRKZ-UHFFFAOYSA-N 3-[4-(aminomethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy-N-(2H-tetrazol-5-ylmethyl)benzamide Chemical compound N=1NN=NC=1CNC(C1=CC(=CC=C1)OC1=NC(=CC(=C1)CN)C(F)(F)F)=O VTNULXUEOJMRKZ-UHFFFAOYSA-N 0.000 description 2
- 102100026816 DNA-dependent metalloprotease SPRTN Human genes 0.000 description 2
- MCNAURNYDFSEML-UHFFFAOYSA-N Guaiane Natural products CC1CCC(C(C)=C)C(O)C2=C(C)C(=O)CC12 MCNAURNYDFSEML-UHFFFAOYSA-N 0.000 description 2
- 125000002619 bicyclic group Chemical group 0.000 description 2
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229940126214 compound 3 Drugs 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 229960003530 donepezil Drugs 0.000 description 2
- QAQCPAHQVOKALN-RMEBNNNOSA-N guaiane Chemical compound C1[C@H](C(C)C)CC[C@H](C)[C@@H]2CC[C@H](C)[C@@H]21 QAQCPAHQVOKALN-RMEBNNNOSA-N 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- CSJDCSCTVDEHRN-UHFFFAOYSA-N methane;molecular oxygen Chemical group C.O=O CSJDCSCTVDEHRN-UHFFFAOYSA-N 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 238000004611 spectroscopical analysis Methods 0.000 description 2
- 238000010183 spectrum analysis Methods 0.000 description 2
- BFDGJIBEQVHHLT-UHFFFAOYSA-N 2-methylprop-1-ene-1,3-diol Chemical compound OCC(C)=CO BFDGJIBEQVHHLT-UHFFFAOYSA-N 0.000 description 1
- 238000005084 2D-nuclear magnetic resonance Methods 0.000 description 1
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 1
- 206010007247 Carbuncle Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 101710175461 DNA-dependent metalloprotease SPRTN Proteins 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 208000018737 Parkinson disease Diseases 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 206010042674 Swelling Diseases 0.000 description 1
- 208000024780 Urticaria Diseases 0.000 description 1
- ABRVLXLNVJHDRQ-UHFFFAOYSA-N [2-pyridin-3-yl-6-(trifluoromethyl)pyridin-4-yl]methanamine Chemical compound FC(C1=CC(=CC(=N1)C=1C=NC=CC=1)CN)(F)F ABRVLXLNVJHDRQ-UHFFFAOYSA-N 0.000 description 1
- OBOXTJCIIVUZEN-UHFFFAOYSA-N [C].[O] Chemical group [C].[O] OBOXTJCIIVUZEN-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 238000005284 basis set Methods 0.000 description 1
- 150000001555 benzenes Chemical class 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000002884 conformational search Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 230000000816 effect on animals Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000005281 excited state Effects 0.000 description 1
- 125000002749 guaiane group Chemical group 0.000 description 1
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 1
- 238000004896 high resolution mass spectrometry Methods 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 230000000324 neuroprotective effect Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 125000004430 oxygen atom Chemical group O* 0.000 description 1
- 125000005704 oxymethylene group Chemical group [H]C([H])([*:2])O[*:1] 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 235000019633 pungent taste Nutrition 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 201000003068 rheumatic fever Diseases 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/10—Spiro-condensed systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C49/00—Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
- C07C49/587—Unsaturated compounds containing a keto groups being part of a ring
- C07C49/703—Unsaturated compounds containing a keto groups being part of a ring containing hydroxy groups
- C07C49/723—Unsaturated compounds containing a keto groups being part of a ring containing hydroxy groups polycyclic
- C07C49/727—Unsaturated compounds containing a keto groups being part of a ring containing hydroxy groups polycyclic a keto group being part of a condensed ring system
- C07C49/733—Unsaturated compounds containing a keto groups being part of a ring containing hydroxy groups polycyclic a keto group being part of a condensed ring system having two rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C49/00—Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
- C07C49/587—Unsaturated compounds containing a keto groups being part of a ring
- C07C49/703—Unsaturated compounds containing a keto groups being part of a ring containing hydroxy groups
- C07C49/723—Unsaturated compounds containing a keto groups being part of a ring containing hydroxy groups polycyclic
- C07C49/727—Unsaturated compounds containing a keto groups being part of a ring containing hydroxy groups polycyclic a keto group being part of a condensed ring system
- C07C49/737—Unsaturated compounds containing a keto groups being part of a ring containing hydroxy groups polycyclic a keto group being part of a condensed ring system having three rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C49/00—Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
- C07C49/587—Unsaturated compounds containing a keto groups being part of a ring
- C07C49/703—Unsaturated compounds containing a keto groups being part of a ring containing hydroxy groups
- C07C49/743—Unsaturated compounds containing a keto groups being part of a ring containing hydroxy groups having unsaturation outside the rings, e.g. humulones, lupulones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2602/00—Systems containing two condensed rings
- C07C2602/02—Systems containing two condensed rings the rings having only two atoms in common
- C07C2602/14—All rings being cycloaliphatic
- C07C2602/26—All rings being cycloaliphatic the ring system containing ten carbon atoms
- C07C2602/30—Azulenes; Hydrogenated azulenes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2603/00—Systems containing at least three condensed rings
- C07C2603/56—Ring systems containing bridged rings
- C07C2603/58—Ring systems containing bridged rings containing three rings
- C07C2603/76—Ring systems containing bridged rings containing three rings containing at least one ring with more than six ring members
- C07C2603/78—Ring systems containing bridged rings containing three rings containing at least one ring with more than six ring members containing seven-membered rings
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- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Neurosurgery (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Psychiatry (AREA)
- Hospice & Palliative Care (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)
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- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
A guaiane sesquiterpene compound separated from Daphne roots, a preparation method and application thereof belong to the technical field of medicines, and in particular relates to 13 guaiane sesquiterpene compounds extracted and separated from Daphne roots (Daphne genkwa Sieb. Et Zucc.) of Daphne, which have the same guaiane sesquiterpene mother nucleus, good acetylcholinesterase inhibition activity and medical application of resisting Alzheimer disease. The preparation method is simple and easy to implement, good in reproducibility and high in purity.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a guaiane sesquiterpene compound prepared from plant lilac daphne roots, a preparation method thereof and application of the compound in the aspect of resisting Alzheimer's disease.
Background
The root of the Daphne genkwa (Daphne genkwa Sieb. Et Zucc.) belonging to the Daphne family (Thymelaeeaceae) is a common traditional Chinese medicine which has long drug-taking history, mild nature, pungent and bitter taste, is flat and toxic, has the effects of clearing heat and detoxicating, relieving cough and asthma and detumescence and diuresis, and is distributed in the regions of each province of Yangtze river and the yellow river basin. It is often used for treating carbuncle, sore, swelling, urticaria, edema, rheumatic arthritis, and rheumatoid arthritis. Pharmacological studies show that the lilac daphne root has certain antibacterial effect, anti-inflammatory effect, neuroprotective activity and anti-tumor effect, and the carbonic acid dissolved part has obvious excitation effect on animal uterus.
With the increasing aging of the global population, neurodegenerative diseases are receiving much attention, and Alzheimer's disease, parkinson's disease and the like become main killers leading to death of middle aged and elderly people. Recent researches show that inhibiting acetylcholinesterase can delay the progress of neurodegenerative diseases, can prevent neurons from being diseased, and has good prospect for treating neurodegenerative diseases.
Disclosure of Invention
The invention aims to provide 13 guaiane sesquiterpenoids separated from Daphne genkwa Sieb.et Zucc root of Daphne genus of daphnaceae family, and the structures are as follows:
the guaiane sesquiterpenoids are extracted and separated from flos Genkwa, which is dry root bark of flos Genkwa of daphne of daphnaceae.
The preparation method of the guaiane sesquiterpene compound comprises the following steps:
extracting dried lilac daphne root with 70% -80% industrial ethanol, mixing the extracting solutions, concentrating to obtain an extract, extracting the extract with ethyl acetate, subjecting the obtained components to silica gel column chromatography, performing isocratic gradient elution with a dichloromethane-methanol system of 100:0-0:100, and collecting three fractions A-C.
And (3) performing gradient elution on the fraction B through HP20 column chromatography by using an ethanol-water system of 0:100-100:0 to obtain four components B1, B2, B3 and B4. And (3) performing gradient elution on the component B1 by using an ODS column chromatography in an ethanol-water system of 10:90-90:10 to obtain five components a-e.
The component b obtained was subjected to silica gel column chromatography with a petroleum ether-ethyl acetate system of 100:1-10:1 to give four subfractions b1-b4 on the basis of TLC analysis.
The mobile phases of methanol-water (70:30, v/v) were used to separate b1 and b2 on preparative reverse phase high performance liquid chromatography to give compounds 1-13.
In the preparation method, the extraction is reflux extraction for 2-3 times, each time for 2-3 hours.
The flos Genkwa is the root of flos Genkwa (Daphne genkwa Sieb. Et Zucc.) of Daphne of daphnaceae.
The results of the system structure identification of the obtained compound are as follows:
the structural identification of compounds 1-13 was performed using high resolution mass spectrometry, one-dimensional NMR, two-dimensional NMR, computational ECD, and NMR techniques (FIGS. 1-26).
Daphnegenol A (1) is yellow oil (methanol) and is easy to dissolve in solvents such as methanol and DMSO.(c 0.10 MeOH) HRESIMS gives an excimer ion peak m/z 289.1415[ M+Na ]] + (calcd.for C 15 H 22 O 4 Na 289.1410), combined with 1 H and 13 c NMR determines that its molecular formula is C 15 H 22 O 4 The unsaturation was calculated to be 5.
1 H NMR(600MHz,DMSO-d 6 ) In delta H 1.62 (3H, s, H-15), 1.14 (3H, s, H-14), 1.08 (3H, s, H-12) are three methyl signals, δ H 3.56 (1H, dd, J=10.5, 4.5Hz, H-13 a), 3.11 (1H, dd, J=10.5, 4.8Hz, H-13 b) is a pair of magnetically unequal CH 2 Upper gem hydrogen signal, delta H 4.40 (1 h, s, oh-10), 4.73 (1 h, t, j=5.2 hz, oh-13) is an active hydrogen signal. 13 C NMR(150MHz,DMSO-d 6 ) The spectra showed 15 carbon signals in total, and in combination with HSQC spectrum analysis, it was speculated that the compound included 1 carbonyl carbon signal delta C 203.9 (C-3), 2 olefin carbon signals delta C 136.2 (C-4), 166.9 (C-5), 4 carbon monoxide signals delta C 83.9 (C-1), 75.5 (C-10), 76.6 (C-11), 66.1 (C-13), 1 methine carbon signal delta C 32.1 (C-7), 4 methylene carbon signals delta C 47.3 (C-2), 24.6 (C-6), 22.3 (C-8), 36.0 (C-9) and 3 methyl carbon signals delta C 24.2(C-12),24.8(C-14),7.8(C-15)。 13 C-NMR(150MHz,CDCl 3 ) There are 29 carbon signals in total, delta C 176.5 is suggested as carbonyl carbon signal, delta C 148.8,144.9,136.5,132.5,132.1,131.9,131.6,124.5,123.8,122.2,117.4,111.4 is 12 sp 2 The hybridized carbon signal, combined with its hydrogen spectrum, is presumed to contain a tetra-substituted benzene ring and three double bonds (including a cis double bond) in the structure. The high field region gives 16 aliphatic carbon signals. The structure is further determined by two-dimensional spectral data analysis.
Delta from HMBC spectra H 2.70,1.85 (H-2) and delta C 83.9 (C-1), 203.9 (C-3), 136.2 (C-4), 166.9 (C-5), which indicates a five-membered ring in the structure. Delta H 2.75,2.50 (H-6) and delta C 83.9 (C-1), 166.9 (C-5), 136.2 (C-4), 76.6 (C-11) and delta H 1.55,1.55 (H-8) and delta C 24.6 (C-6), 32.1 (C-7), 36.0 (C-9), 75.5 (C-10) and the like, which confirm that the compound 1 is a 5/7 bicyclic skeleton. Delta H 1.08 (H-12) and delta C 76.6 (C-11), 32.1 (C-7), 66.1 (C-13) the correlation states that C-11 is linked to C-7, C-12, C-13, which constitutes a skeleton of the guaiane-type sesquiterpene. Delta H 1.14 (H-14) and delta C 83.9 (C-1), 75.5 (C-10), 36.0 (C-9) and delta H 1.62 (H-15) and delta C 203.9 (C-3), 136.2 (C-4), 166.9 (C-5) and it was determined that two methyl groups were attached at positions 10 and 4, respectively. From HMBC signal delta H 4.40 (OH-10) delta C 83.9(C-1),75.5(C-10), 36.0 (C-9), 24.8 (C-14), 4.73 (OH-13) and delta C 66.1 (C-13), 76.6 (C-11) has defined two hydroxyl groups attached at positions 10 and 13, respectively. The planar configuration of the compound 1 is determined by the fact that the compound 1 has 5 unsaturations, the guaiane skeleton occupies 2 unsaturations, the double bond and the carbonyl group occupy 1 unsaturation, and one unsaturation is presumably obtained by connecting C-1 and C-11 through oxygen atoms to form a ring.
The relative configuration of compound 1 was determined by NOESY spectra and X-single crystal diffraction. The presence of NOE in H-12/H-6β correlates with the presence of NOE in H-14/H-13, indicating that Me-12 and H-6β are on the same side, and H-13 is on the same side as Me-14, combined with X-single crystal diffraction results, thereby determining the relative and absolute configuration of the compounds. The absolute configuration of the compound was verified by calculation of ECD and comparison with the measured ECD by first performing a conformational search using Spartan 14.0 under MMFF94 force field, selecting a conformation with Boltzmann distribution greater than 1% optimized at the B3LYP/6-31G (d) basal group level, then further performing an excited state calculation at the B3LYP/6-311++ G (2 d, p) basal group level, and finally fitting the ECD spectrum using SpecDis software. Compound 1 conformed well to the calculated 1r,7r,10r,11s configuration, thereby determining the absolute configuration of the compound.
daphnegenol B (2) is a yellow oil (methanol) which is easily dissolved in solvents such as methanol and DMSO.(c 0.10 MeOH) HRESIMS gives an excimer ion peak m/z 289.1412[ M+Na ]] + (calcd for C 15 H 22 O 4 Na, 289.1410), combined with 1 H-NMR 13 C-NMR deduces that its molecular formula is C 15 H 22 O 4 The unsaturation was calculated to be 5.
1 H NMR(600MHz,DMSO-d 6 ) The spectrum shows 1 olefinic carbon signal delta H 5.96 (1 h, d, j=1.2 hz, h-9), 3 active hydrogen signals δ H 4.77 (1H, s, OH-7), 4.38 (1H, o, OH-4), 4.37 (1H, o, OH-13); 2 methyl signals delta H 2.02(3H,d,J＝1.2Hz,H-14),1.17(3H,s,H-15)。 13 C NMR(150MHz,DMSO-d 6 ) Co-spectrum15 carbon signals were shown, combined with HSQC spectrum analysis, and it was speculated that 1 carbonyl carbon signal delta was included in the compound C 202.1 (C-8), 2 olefin carbon signals delta C 172.7 (C-10), 127.5 (C-9), 3 carbon oxygen signals delta C 78.6 (C-4), 81.0 (C-7), 62.0 (C-13), 1 sp 3 Hybrid quaternary carbon signal delta C 50.9 (C-1), 2 methine carbon signals delta C 56.9 (C-5), 45.0 (C-11), 5 methylene carbon signals delta C 32.2 (C-2), 39.9 (C-3), 33.6 (C-6), 30.5 (C-12) and 2 methyl carbon signals delta C 22.3(C-14),26.3(C-15)。
According to 1 H- 1 H COSY spectrum, compound 2 has two spin-coupled systems, delta in HMBC spectrum H 2.13 (H-2) and delta C 50.9 (C-1), 172.7 (C-10), 78.6 (C-4), 30.5 (C-12) are related, delta H 1.40 (H-2) and delta C 39.9 (C-3), 56.9 (C-5), 78.6 (C-4) are related, delta H 1.68 (H-3) and delta C 32.2 (C-2), 50.9 (C-1), 78.6 (C-4) are present to define five-membered ring fragments. Delta H 5.96 (H-9) and delta C 22.3 (C-14), 50.9 (C-1), 81.0 (C-7), 172.7 (C-10), 1.64 (H-6) and delta C 81.0 (C-7), 202.1 (C-8) being present in relation to 1.51 (H-6) and delta C 50.9 (C-1), 81.0 (C-7), 56.9 (C-5), 202.1 (C-8) are present to confirm that the compound contains a seven-membered ring fragment. Delta H 1.17 (H-15) and delta C 39.9 (C-3), 78.6 (C-4), 56.9 (C-5) are related, delta H 2.02 (H-14) and delta C 50.9 (C-1), 127.5 (C-9), 172.7 (C-10) are correlated, thereby determining the positions of the two methyl groups, delta H 4.38 (H-4) and delta C 39.9 (C-3), 78.6 (C-4), 56.9 (C-5), 26.3 (C-15) are present in relation, delta H 4.77 (H-7) and delta C 33.6 (C-6), 81.0 (C-7), 202.1 (C-8), 45.0 (C-11) are correlated, delta H 4.37 (OH-13) delta C 45.0 (C-11), 62.0 (C-13), positions of three hydroxyl groups, delta, are determined H 2.91 (H-13) and delta C 30.5 (C-11), 45.0 (C-12) and a hydroxymethyl group attached at position 11 are determined, thereby determining the planar structure of the compound 2.
The relative configuration of compound 2 was determined by both NOESY spectra and calculated nuclear magnetism. There is a correlation between H-5/H-9,H-9/H-13, indicating that H-5 is linked to a carbon bridgeC-1/C10/C9/C8/C7 is on the same side, and H-13 is on the same side as H-9. After optimization of conformations with Boltzmann distribution greater than 1% at the B3LYP/6-31G (d) basis set level using Spartan software under MMFF force field, further calculations were performed at the mPW PW91/6-311+g (d, p) basis set level to calculate two possible conformations (1R, 4R,5R,7S, 11R) -2 and (1R, 4S, 5R,7S, 11R) -2 13 C-NMR 1 H NMR, and finally, nuclear magnetic data of two configurations are calculated by using a weighted summation mode, and the relative configuration of the compound 2 is 1R,4R,5R,7S and 11R in the case that the dp4+ probability is close to 100%. The ECD spectra of 1R,4R,5R,7S,11R-2 and 1S,4S,5S,7R,11S-2 were calculated, and the absolute configuration was determined to be 1R,4R,5R,7S,11R by comparison of the experimental and calculated ECD spectra.
Table 1 compounds 1 and 2 1 H (600 MHz) and 13 c (150 MHz) NMR data (DMSO-d 6 )
daphnegenol C (3) colorless crystals (methanol), which are readily soluble in solvents such as methanol and DMSO.(c 0.10 MeOH) HRESIMS gives an excimer ion peak m/z 289.1415[ M+Na ]] + (calcd for C 15 H 22 O 4 Na, 289.1410), combined with 1 H-NMR 13 C-NMR deduces that its molecular formula is C 15 H 22 O 4 The unsaturation was calculated to be 5.
1 H NMR(600MHz,DMSO-d 6 ) The spectrum shows 3 hydroxyl hydrogen proton signals delta H 5.18 (1H, s, OH-7), 4.73 (1H, s, OH-8), 4.71 (1H, dd, J=5.6, 2.1Hz, OH-13), three methyl hydrogen signals δ H 0.83(3H,s,H-12),1.16(3H,s,H-14),1.07(3H,d,J＝7.1Hz,H-15)。 13 C NMR(150MHz,DMSO-d 6 ) The spectra showed 15 carbon signals in total, which are presumed to be sesquiterpenoids, and in combination with HSQC spectral analysis, 1 alpha, beta-unsaturated ketone signal delta is presumed to be included in the compounds C 204.9 (C-2), 176.5 (C-5), 143.6 (C-1), 3 carbon monoxide signals delta C 83.5 (C-7), 76.8 (C-8), 64.2 (C-13), 2 sp 3 Hybrid quaternary carbon signal delta C 43.6 (C-2), 49.2 (C-2), 1 methine carbon signal delta C 33.7 (C-2), 3 methylene carbon signals delta C 46.6 (C-9), 44.7, (C-3) 33.9 (C-6) and 3 methyl carbon signals delta C 18.2(C-2),14.1(C-2),12.3(C-2)。
Delta in HMBC spectra H 2.47 (H-3) and delta C 204.9 (C-2), 176.5 (C-5), 143.6 (C-1), 33.7 (C-4) and 18.2 (C-15) are present in relation, delta H 2.60 (H-1) and delta C 176.5 (C-5), 143.6 (C-1), 44.7 (C-3) and 18.2 (C-15) are present in relation to each other, indicating that it is a five membered ring structure containing an alpha, beta-unsaturated ketone fragment. Delta H 1.07 (H-15) and delta C 176.5 (C-5), 44.7 (C-3) and 33.7 (C-4) are related, indicating that C-15 is attached to C-4. Delta H 2.67 (H-6) and delta C 176.5 (C-5), 143.6 (C-1), 83.5 (C-7), 76.8 (C-8), 49.2 (C-11) and 33.7 (C-4) are present in relation, delta H 2.14 (H-9) and delta C 143.6 (C-1), 43.6 (C-10), which indicates the presence of a seven-membered ring, delta H 0.83 (H-12) and delta C 83.5 (C-7), 64.2 (C-13), 43.6 (C-10), 49.2 (C-11) and delta H 3.79 (H-13) and delta C 83.5 (C-7), 49.2 (C-11), 43.6 (C-10) and 12.3 (C-12) are related, indicating that C-11 is linked to C-10, C-7, whereby the planar structure of the compound is determined.
From the NOESY spectra, H-15 correlated with H-6β, H-6β correlated with H-12, combined with X-single crystal diffraction, the absolute configuration of compound was determined to be 4S,7S,10R,11R. The absolute configuration of this compound is 4s,7s,10r,11r.
daphnegenol D (4) is a yellow oil (methanol) which is easily dissolved in solvents such as methanol and DMSO.(c 0.10MeOH)，HRESIMS gives an excimer ion peak m/z 289.1412[ M+Na ]] + (calcd for C 15 H 22 O 4 Na, 289.1410), combined with 1 H-NMR 13 C-NMR deduces that its molecular formula is C 15 H 22 O 4 The unsaturation was calculated to be 5.
1 H NMR(600MHz,DMSO-d 6 ) In the spectrum, delta H 5.64 (1H, s, H-2) is an olefinic carbon signal, delta H 5.05 (1 h, d, j=3.9 hz, 8-OH), 5.01 (1 h, s, 7-OH), 4.80 (1 h, s, 13-OH) signals are 3 active hydrogen proton signals, δ H 3.78 (1 h, dd, j=11.1, 2.3hz, h-13 a), 3.45 (1 h, dd, j=11.1, 5.5hz, h-13 b), 1.87 (1 h, dd, j=12.1, 8.7hz, h-6α), 1.49 (1 h, dd, j=12.1, 12.1hz, h-6β) are two pairs of geminally coupled methylene signals. 13 C NMR(150MHz,DMSO-d 6 ) A total of 15 carbon signals are displayed in the spectrum, and delta is combined with analysis of HSQC spectrum C 211.3 (C-3), 190.7 (C-1), 122.4 (C-2) is an alpha, beta-unsaturated ketone signal, delta C 83.1 (C-7), 75.1 (C-8), 64.5 (C-13) the hint structure contains 3 oxygen-linked sp 3 Hybrid carbon, delta C 50.2 (C-11), 46.4 (C-10), 42.9 (C-9), 42.6 (C-4), 38.3 (C-5), 30.8 (C-6) indicate 6 sp in the structure 3 Hybrid carbon, delta C 17.1 (C-14), 13.0 (C-12), and 12.6 (C-15) suggest that there are 3 methyl groups in the structure.
In HMBC spectra, delta H 5.63 (H-2) and delta C 211.3 (C-3), 190.7 (C-1), 46.4 (C-10), 42.6 (C-4), 38.3 (C-5), indicating that the structure contains a five-membered ring, delta H 4.33 (H-8) and delta C 83.1 (C-7), 46.4 (C-10), 42.9 (C-9), 36.8 (C-6) are related, indicating that the structure contains a seven-membered ring, delta H 0.66 (H-12) and delta C 50.2 (C-11), 64.5 (C-13), 46.4 (C-10), 83.1 (C-7), proving that the C-11 position is linked to C-7, C-10 and C-12, delta H 1.04 (H-14) and delta C 190.7 (C-1), 50.2 (C-11), 42.9 (C-9) are related, delta H 0.96 (H-15) and delta C 211.3 (C-3), 38.3 (C-5) positions the other two methyl groups, and is combined with 1 H- 1 The H COSY spectrum shows 3 spin systems, which ultimately determine the planar structure of compound 4.
The relative configuration of compound 4 was determined by NOESY spectroscopy. H-14 and H-12 are correlated, H-12 and H-15 are correlated, H-5 and H-9α are correlated, H-9β and H-14 are correlated, and the relative configuration of compound 4 is determined to be 4R, 5S,7S,8R,10S,11R by combining the calculated nuclear magnetism and the DP4+ analysis. The absolute configuration of compound 4 was determined by comparing the calculated ECD with the measured ECD, thereby determining that the absolute configuration of compound 4 was 1r,7r,10s.
TABLE 2 Compounds 3 and 4 1 H (600 MHz) and 13 c (150 MHz) NMR data (DMSO-d 6 )
daphnegenol E (5) is a yellow oil (methanol) which is easily dissolved in solvents such as methanol and DMSO.(c 0.10 MeOH) HRESIMS gives an excimer ion peak 289.1411[ M+Na ]] + (calcd for C 15 H 22 O 4 Na, 289.1410), the molecular formula of the compound is C 15 H 22 O 4 The unsaturation was 5.
1 H NMR(600MHz,DMSO-d 6 ) The spectrum shows 3 hydroxyl hydrogen signals delta H 4.99 (1H, s, OH-7), 4.99 (1H, s, OH-8), 4.80 (1H, br.s, OH-13); 3 methyl proton signals delta H 0.67 (3H, s, H-12), 1.03 (3H, s, H-14), 1.04 (3H, o, H-15); hydrogen signal delta of 1 double bond H 5.64 (1 h, d, j=1.8 hz, h-2); group 1 methylene hydrogen signals 3.79 (1 h, d, j=11.1 hz, h-13 a), 3.44 (1 h, d, j=11.1 hz, h-13 b). 13 C NMR(150MHz,DMSO-d 6 ) The spectra showed 15 carbon signals in total, and in combination with HSQC spectrum analysis, it was speculated that the compound included 1 carbonyl carbon signal delta C 209.4 (C-3), 2 double bond carbon signals delta C 189.4 (C-1), 122.6 (C-2), 3 oxygen-linked aliphatic carbon signals delta C 83.0 (C-7), 75.0 (C-8), 64.6 (C-13), 6 fatsCarbon signal delta C 49.1 (C-4), 43.7 (C-5), 35.3 (C-6), 42.7 (C-9), 46.1 (C-10), 50.3 (C-11) and 3 methyl carbon signals delta C 13.4(C-12),17.0(C-14),13.6(C-15)。
Delta in HMBC spectra H 5.64 (H-2) and delta C 209.4 (C-3), 189.4 (C-1), 46.1 (C-10), 49.1 (C-4), 43.7 (C-5), indicating that the structure contains a five-membered ring, delta H 4.33 (H-8) and delta C 83.0 (C-7), 46.1 (C-10), 42.7 (C-9), 35.3 (C-6) are related, indicating that the structure contains a seven-membered ring, delta H 0.67 (H-12) and delta C 50.3 (C-11), 64.6 (C-13), 46.1 (C-10), 83.0 (C-7), proving that the C-11 position is linked to C-7, C-10 and C-12, delta H 1.03 (H-14) and delta C 189.4 (C-1), 50.3 (C-11), 42.7 (C-9) are related, delta H 1.04 (H-15) and delta C 209.4 (C-3), 43.7 (C-5) determines the position of the other two methyl groups, in combination with it 1 H- 1 The H COSY spectra are related and the following fragments can be ligated, thus determining the planar structure of Compound 5.
The relative configuration of compound 5 was determined from the NOESY spectrum and calculated nuclear magnetism. The relative configuration of the compounds was determined to be 4S,5S,7S,8R,10S,11R by combining calculated nuclear magnetism with DP4+ analysis, with the NOE correlation of H-15/H-5,H-5/H-9α, H-9β/H-14, H-12/H-14, H-9β/H-13. The absolute configuration of this compound is determined by comparing the measured ECD with the calculated ECD, so the absolute configuration of this compound is 4S,5S,7S,8R,10S,11R.
daphnegenol F (6) is a yellow oil (methanol) which is easily dissolved in solvents such as methanol and DMSO.(c 0.10 MeOH). HRESIMS gives an excimer ion peak of 305.1394[ M+Na ]] + (calcd for C 15 H 22 O 5 Na, 305.1359), the molecular weight of the compound was 282, and the molecular formula was C 15 H 22 O 5 The calculated unsaturation was 5.
1 H NMR(600MHz,DMSO-d 6 ) In the spectrum, delta H 2.09 (3 h, d, j=2.5 hz, h-14), 0.72 (3 h, d, j=7.1 hz, h-15) are two methyl signals; delta H 4.38(1 h, s, oh-11), 4.46 (1 h, o, oh-13), 5.10 (1 h, d, j=4.6 hz, oh-8) are 3 active hydrogen signals. 13 C NMR(150MHz,DMSO-d 6 ) The spectra showed 15 carbon signals in total, combined with HSQC spectral analysis, and it was inferred that the compound included 1 carbonyl carbon signal delta C 207.7 (C-2), 2 ethylenic carbon signals delta C 138.0 (C-1), 143.0 (C-10), 5 carbon monoxide signals delta C 67.2 (C-8), 81.1 (C-9), 75.0 (C-11), 64.2 (C-12), 62.4 (C-13), 3 methine carbon signals delta C 30.6 (C-4), 39.1 (C-7), 39.4 (C-5), 2 methylene signals delta C 29.4 (C-6), 48.7 (C-3), 2 methyl carbon signals delta C 15.6(C-15)，18.5(C-14)。
Delta in HMBC spectra H 2.45 (H-3) and delta C 30.6 (C-4), 15.6 (C-15), 207.7 (C-2), 138.0 (C-1), 39.4 (C-5) are correlated, delta H 2.14 (H-4) and delta C 207.7 (C-2), 138.0 (C-1) is present, indicating the presence of a five-membered ring fragment. Delta H 0.72 (H-15) and delta C 48.7 (C-3), 39.4 (C-5), 30.6 (C-4) are present to indicate that C-15 is attached to C-4. Delta H 1.73 (H-6) and delta C 39.1 (C-7), 75.0 (C-11) presence-related, delta H 4.46 (H-8) and delta C 143.0 (C-10), 81.1 (C-9), 39.1 (C-7), 29.4 (C-6) are present in relation, delta H 4.06 (H-9) and delta C 143.0 (C-10), 138.0 (C-1), 67.2 (C-8), 62.4 (C-12), 39.1 (C-7), 18.5 (C-14) are related, indicating the presence of a seven-membered ring. Delta H 3.32 (H-12) and delta C 81.1 (C-9), 75.0 (C-11), 39.1 (C-7) are present in relation to C-9 (delta) C 81.1),C-12(δ C 62.4 Chemical shift values, indicating the presence of six-membered oxygen ring structural fragments. Delta H 2.09 (H-14) and delta C 207.7 (C-2), 143.0 (C-10), 138.0 (C-1), 81.1 (C-9) are present in relation, indicating that C-14 is attached to C-10. Delta H 3.31 (H-13) and delta C 39.1 (C-7), 62.4 (C-12), 75.0 (C-11) are present to indicate that C-13 is attached to C-11. The planar structure of compound 6 can be determined therefrom.
The relative configuration of compound 6 was determined by NOESY spectra together with calculated nuclear magnetism. NOE correlation exists between H-7/H-9,H-7/OH-8,H-9/OH-8, OH-8/H-5,H-13/H-6. Calculating the nuclear magnetism of (4S, 5S,7S,8R,9R, 11S) -6 and (4R, 5S,7S,8R,9R, 11S) -6 to determine the relative configuration of the compound 6 as 4S,5S,7S,8R,9R,11S. The absolute configuration of this compound is determined by comparing the measured ECD with the calculated ECD, so the absolute configuration of this compound is 4S,5S,7S,8R,9R,11S.
TABLE 3 Compounds 5 and 6 1 H (600 MHz) and 13 c (150 MHz) NMR data ((DMSO-d) 6 )
daphnegenol G (7) is a yellow oil (methanol) which is easily dissolved in solvents such as methanol and DMSO.(c 0.10 MeOH) HRESIMS gives an excimer ion peak m/z 291.1525[ M+Na ]] + (calcd for C 15 H 24 O 4 Na, 291.1567), combined with 1 H NMR 13 C NMR deduces that its molecular formula is C 15 H 24 O 4 The unsaturation was calculated to be 4.
1 H NMR(600MHz,DMSO-d 6 ) The spectrum shows 3 active hydrogen signals delta H 5.20 (1H, s, 10-OH), 4.42 (1H, s, 13-OH), 3.99 (1H, s, 11-OH); 3 methyl signals delta H 0.95(3H,s,H-12),1.56(3H,s,H-15),0.61(3H,d,J＝7.0Hz,H-14)。 13 C NMR(150MHz,DMSO-d 6 ) The spectra showed 15 carbon signals in total, and in combination with HSQC spectrum analysis, it was speculated that the compound included 1 carbonyl carbon signal delta C 204.3 (C-3), 2 ethylenic carbon signals delta C 173.2 (C-5), 136.3 (C-4), 3 carbon oxygen signals delta C 81.3 (C-1), 73.9 (C-11), 67.6 (C-13), 2 methine carbon signals delta C 39.2 (C-10), 41.2 (C-7), 4 methylene carbon signals delta C 23.6 (C-8), 30.6 (C-6), 31.0 (C-9), 50.7 (C-2) and 3 methyl carbon signals delta C 20.3(C-12),14.4(C-14),7.6(C-15)。
By passing through 1 H- 1 The H COSY spectrum shows that the compound 7 has a spin coupling system according to delta in the HMBC spectrum H 2.42 (H-2) and delta C 81.2(C-1),204.3(C-3),39.2(C-10),136.3(C-4)，δ H 1.56 (H-15) and delta C 204.3 (C-3), 136.3 (C-4), 173.2 (C-5) are present in association with 13 C NMR confirms that the compound contains a five membered ring fragment, C-15 is attached to C-4. Delta H 1.35 (H-9) and delta C 81.2 (C-1), 39.2 (C-10), 31.0 (C-9), 23.6 (C-8) and 41.2 (C-7) are present in relation, delta H 2.65 (H-6) and delta C 23.6 (C-8) there is a correlation, and the structure of the seven-membered ring is determined. Delta H 5.20 (OH-1) and delta C 40.4(C-10),δ C 81.3(C-1),δ C 173.2 (C-5) there is a correlation; delta H 3.99 (OH-11) delta C 73.9(C-11),δ C 67.6(C-13),δ C 20.3 (C-12) there is a correlation; delta H 3.30 (OH-13) delta C 20.3(C-12),δ C 73.9 (C-11) there is correlation, determining the positions of three hydroxyl groups; delta H 0.95 (H-12) and delta C 41.1(C-7)，δ C 73.9(C-11),δ C 67.6 (C-13) the related description methyl group was attached to C-12, whereby the planar structure of Compound 7 was determined.
The relative configuration of compound 7 was determined by NOESY spectroscopy. The presence of H-2α and H-14 correlates, and the presence of H-2β and OH-1 correlates, indicating that Me-14 and OH-1 are off-plane. The relative configuration was determined by calculation of nuclear magnetism and dp4+ analysis to be 1R,7R,10S, 11S. The absolute configuration of the compound is determined by comparing the measured ECD with a method for calculating the ECD, and the calculated 1R,7R,10S,11S configuration has higher map matching degree with the measured ECD, so that the absolute configuration of the compound is determined to be 1'S, 1' R,4 'S, 6' R.
daphnegenol H (8) is a yellow oil (methanol) which is easily dissolved in solvents such as methanol and DMSO.(c 0.10 MeOH) HRESIMS gives an excimer ion peak m/z 289.1415[ M+Na ]] + (calcd for C 15 H 22 O 4 Na, 289.1410), combined with 1 H-NMR 13 C-NMR deducing its molecular formulaIs C 15 H 22 O 4 The unsaturation was calculated to be 5.
1 H NMR(600MHz,DMSO-d 6 ) The spectrum shows 3 active hydrogen signals delta H 5.25 (1 h, s, oh-1), 4.80 (1 h, t, j=5.5 hz, oh-13), 4.45 (1 h, s, oh-10); 2 methyl signals delta H 0.82 (3H, s, H-14), 1.53 (3H, s, H-15); proton signal delta of terminal double bond H 4.96(1H,d,J＝1.6Hz,H-12a),4.84(1H,d,J＝1.6Hz,H-12b)。 13 C NMR(150MHz,DMSO-d 6 ) The spectra showed 15 carbon signals in total, and in combination with HSQC spectrum analysis, it was speculated that the compound included 1 carbonyl carbon signal delta C 205.2 (C-3), 4 olefin carbon signals delta C 171.5 (C-5), 136.4 (C-4), 155.9 (C-11), 106.5 (C-12), 3 carbon monoxide signals delta C 84.4 (C-1), 75.2 (C-10), 63.0 (C-13), 1 methine carbon signal delta C 36.9 (C-7), 4 methylene carbon signals delta C 48.1 (C-2), 36.4 (C-6), 31.1 (C-8), 39.5 (C-9) and 2 methyl carbon signals delta C 22.5(C-14),7.5(C-15)。
According to 1 H- 1 The H COSY spectrum, compound 8, shows a spin-coupling system. Delta in HMBC spectra H 2.13 (H-2) and delta C 75.2 (C-10), 84.4 (C-1), 136.4 (C-4), 171.5 (C-5), 205.3 (C-3) are present in relation to determine that the compound contains a five-membered ring. Delta H 2.84 (H-7) and delta C 31.3 (C-8), 36.4 (C-6), 63.0 (C-13), 106.5 (C-12), 155.9 (C-11) are related, delta H 1.29 (H-8) and delta C 36.9 (C-7), 39.5 (C-9), 75.2 (C-10), 155.9 (C-11) are related, indicating the presence of structural fragments of the seven-membered ring. Delta H 0.82 (H-14) and delta C 39.5 (C-9), 75.2 (C-10), 155.9 (C-11) are related, delta H 1.53 (H-15) and delta C 136.4 (C-4), 171.5 (C-5), 205.3 (C-3) are correlated, thereby determining the positions of the two methyl groups, delta H 5.25 (OH-1) and delta C 75.2 (C-10), 84.4 (C-1), 48.1 (C-2), 171.5 (C-5) are present in relation, delta H 4.45 (OH-10) delta C 84.4 (C-1), 39.5 (C-9), 75.2 (C-10), 22.5 (C-14) are present in relation, delta H 4.80 (OH-13) delta C 155.9 (C-11), 63.0 (C-13) are correlated, and the positions of three hydroxyl groups are determined; delta H 4.84 (H-12) and delta C 36.9 (C-7), 155.9 (C-11), 63.0 (C-13) are related, delta H 3.94 (H-13) and delta C 36.9 (C-7), 155.9 (C-11), 106.5 (C-12) the side chain in the C-7 position is 3-hydroxy-2-methylpropenol. The planar structure of compound 8 was thus determined.
Determination of its relative configuration as 1R by calculation of the nuclear magnetism and DP4+ analysis * ,7R * ,10S * . There is a correlation between H-7/H-9α, H-9β/H-14 in the NOESY spectrum, which demonstrates the relative configuration of compound 8. Compound 8 conformed well to the calculated 1r,7r,10s configuration, thereby determining the absolute configuration of the compound.
TABLE 4 Compounds 7 and 8 1 H (600 MHz) and 13 c (150 MHz) NMR data (DMSO-d 6 )
daphnegenol I (9) is a yellow oil (methanol) which is easily dissolved in solvents such as methanol and DMSO.(c 0.10 MeOH) HRESIMS gives an excimer ion peak 289.1400[ M+Na ]] + (calcd for C 15 H 22 O 4 Na, 289.1410), the molecular weight of the compound was 266, and the molecular formula was C 15 H 22 O 4 The calculated unsaturation was 5.
1 H NMR(600MHz,DMSO-d 6 ) Delta in spectrum H 1.05 (3 h, d, j=7.3 Hz), 1.58 (3 h, s) is two methyl signals, δ H 4.98 (1 h, d, j=1.6 hz, h-12 a), 4.86 (1 h, d, j=1.6 hz, h-12 b) is a proton signal on a set of terminal double bonds, δ H 5.12 (1 h, s, oh-1), 4.56 (1 h, s, oh-10), 4.82 (1 h, t, j=5.5 hz, oh-13) are three active hydrogen signals. 13 C NMR (150 MHz, DMSO) spectra showed 15 carbon signals in total, combined with HSQC spectraAnalysis, deducing that the structure contains 1 carbonyl carbon signal delta C 206.1 (C-3), 4 olefin carbon signals delta C 173.6 (C-5), 135.5 (C-4), 155.0 (C-11), 106.7 (C-12), 3 carbon monoxide signals delta C 80.7 (C-1), 72.7 (C-10), 62.8 (C-13), 1 methine carbon signal delta C 41.6 (C-7), 4 methylene carbon signals delta C 46.6 (C-2), 30.4 (C-6), 27.9 (C-8), 34.9 (C-9) and 2 methyl carbon signals delta C 25.4(C-14),7.2(C-15)。
Delta in HMBC spectra H 2.53 (H-2) and delta C 206.1 (C-3), 173.6 (C-5), 135.5 (C-4), 80.7 (C-1), 72.7 (C-10) are related, indicating that they are five-membered ring structures in which alpha, beta-unsaturated ketone fragments are present. Delta H 1.58 (H-15) and delta C 206.1 (C-3), 135.5 (C-4), 173.6 (C-5) are present in relation, indicating that C-15 is attached to C-4. Delta H 2.69 (H-6) and delta C 173.6 (C-5), 135.5 (C-4), 80.7 (C-1), 41.6 (C-7), 27.9 (C-8), delta H 1.85 (H-8) and delta C 72.7 (C-10), 41.6 (C-7), delta H 1.90 (H-9) and delta C 80.7 (C-1), 72.7 (C-10), 41.6 (C-7), 25.4 (C-14) are related, indicating the presence of a seven-membered ring fragment. Delta H 1.05 (H-14) and delta C 80.7 (C-1), 72.7 (C-10), 34.9 (C-9) are present in relation, indicating that C-14 is attached to C-10. Delta H 4.98 (H-12) and delta C 41.6 (C-7), 62.8 (C-13), 155.0 (C-11) are related, delta H 3.95 (H-13) and delta C 155.0 (C-11), 106.7 (C-12), 41.6 (C-7) are present to correlate, indicating that C-11 is linked to C-12, C-7, C-13, and by delta C 155.0 (C-11), 106.7 (C-12) can judge that the C-11 and the C-12 are connected through double bonds, and the plane structure of the compound is determined to be the guaiane type sesquiterpene by combining the signals.
The relative configuration is determined by NOESY spectra together with the calculated nuclear magnetism. The presence of NOE in H-7/H-14 correlates to the presence of H-7 and Me-14 in the alpha configuration and OH-10 in the beta configuration. In combination with calculating the nuclear magnetism, the relative configuration of compound 9 was determined to be 1R,7R,10S. The absolute configuration of the compound was determined by comparing the calculated ECD with the measured ECD, and the absolute configuration of the compound was 1R,7R,10S.
Daphnegenol J (10) yellow oil (methanol) soluble in formazanAlcohols, DMSO, and the like.(c 0.10 MeOH) HRESIMS gives an excimer ion peak 289.1407[ M+Na ]] + (calcd for C 15 H 22 O 4 Na, 289.1410), the molecular weight of the compound was 266, and the molecular formula was C 15 H 22 O 4 The calculated unsaturation was 5.
1 H NMR(600MHz,DMSO-d 6 ) In the spectrum, two methyl proton signals δ are given H 1.07 (3 h, s, h-15), 0.97 (3 h, d, j=7.3 hz, h-14), proton signal δ at a set of terminal double bonds H 4.99 (1 h, d, j=1.6 hz, h-12 a), 4.89 (1 h, d, j=1.6 hz, h-12 b) and three hydroxyl proton signals δ H 5.60(1H,d,J＝6.2Hz,OH-3),5.23(1H,s,OH-4),4.86(1H,t,J＝5.0Hz,OH-13)。 13 C NMR(150MHz,DMSO-d 6 ) The spectrum shows 15 carbon signals in total, and can be attributed to one carbonyl carbon signal delta C 202.2 (C-2) and two sets of olefinic carbon signals delta C 171.8 (C-5), 155.4 (C-11), 141.2 (C-1), 106.8 (C-12); three oxygen-carbon signals delta C 81.1 (C-3), 77.9 (C-4), 63.0 (C-13); two methine carbon signals delta C 41.2 (C-7), 25.9 (C-10); three methylene carbon signals delta C 32.5 (C-6), 32.2 (C-9), 30.6 (C-8); two methyl carbon signals delta C 22.4(C-15),16.6(C-14)。
In HMBC spectra, δ H 2.53 (H-3) and delta C 202.2 (C-2), 77.9 (C-4), 22.4 (C-15) are present to illustrate the presence of a five-membered ring structural fragment of an alpha, beta-unsaturated ketone in the compound. Delta H 1.07 (H-15) and delta C 171.8 (C-5), 81.1 (C-3), 77.9 (C-4) are related, delta H 5.60 (OH-3) delta C 202.2 (C-2), 81.1 (C-3), 77.9 (C-4) are related, delta H 5.23 (OH-4) and delta C 171.8 (C-5), 77.9 (C-4), 22.4 (C-15) are present in relation, indicating that C-15 is attached to C-4 and two hydroxyl groups are attached to C-3 and C-4, respectively. Delta H 2.42 (H-6) and delta C 171.8 (C-5), 155.4 (C-11), 141.2 (C-1), 41.2 (C-7), 30.6 (C-8), delta H 1.70 (H-9) and delta C 16.6 (C-14), 41.6 (C-7), indicating the presence of one of the compoundsSeven membered rings. Delta H 0.97 (H-14) and delta C 141.2 (C-1), 32.2 (C-9), 25.9 (C-10) are present in relation, indicating that C-14 is attached to C-10. Delta H 4.89 (H-12) and delta C 155.4 (C-11), 63.0 (C-13), 41.2 (C-7) are related, delta H 3.96 (H-13) and delta C 155.4 (C-11), 106.8 (C-12), 41.2 (C-7) are related, which indicates that C-11 is linked to C-12, C-7, C-13, and the above signals are combined to determine the planar structure of the compound.
The relative configuration is determined by NOESY spectra together with the calculated nuclear magnetism. H-6β is related to the presence of NOE for H-14, indicating that H-6β and Me-14 are in the β configuration; h-6α and H-15 are related to NOE, H-6α and Me-15 are in an α configuration, and OH-4 is in a β configuration. In combination with calculating the nuclear magnetism, the relative configuration of compound 10 was determined to be 3R,4R,7R,10S. Absolute configuration 3r,4r,7r,10s was determined by combining the calculated ECD with the measured ECD.
Table 5 compounds 9 and 10 1 H (600 MHz) and 13 c (150 MHz) NMR data (DMSO-d 6 )
Daphnegenol K (11) is yellow oil (methanol) and is easy to dissolve in solvents such as methanol and DMSO.(c 0.10 MeOH) HRESIMS gives an excimer ion peak 289.1415[ M+Na ]] + (calcd for C 15 H 22 O 4 Na, 289.1410), the molecular formula of the compound is C 15 H 22 O 4 The unsaturation was 5.
1 H NMR(600MHz,DMSO-d 6 ) The spectrum shows 4 hydroxyl hydrogen signals delta H 5.47 (1 h, s, oh-1), 5.28 (1 h, d, j=5.6 hz, oh-8), 4.70 (1 h, s, oh-10), 4.90 (1 h, t, j=5.4 hz, oh-13); 2 methyl proton signals delta H 1.17 (3H, s, H-14), 1.60 (3H, s, H-15); hydrogen signal delta at 1 terminal double bond H 5.00(2H,s)。 13 C NMR(150MHz,DMSO-d 6 ) The spectra showed 15 carbon signals in total, and in combination with HSQC spectrum analysis, it was speculated that the compound included 1 carbonyl carbon signal delta C 205.1 (C-3), 4 double bond carbon signals delta C 136.5 (C-4), 171.8 (C-5), 153.3, (C-11) 108.2 (C-12), 4 oxygen sp3 hybridized carbon signals delta C 83.8 (C-1), 76.1 (C-10), 71.6 (C-8), 63.3 (C-13), 4 sp3 hybridized carbon signals delta C 47.8 (C-2), 29.3 (C-6), 41.5 (C-7), 44.2 (C-9), and 2 methyl carbon signals delta C 27.3(C-14),7.8(C-15)。
Delta in HMBC spectra H 2.71 (H-2) and delta C 205.1 (C-3), 136.5 (C-4), 171.8 (C-5) and 83.8 (C-1) to determine the presence of five-membered ring fragments; delta H 1.60 (H-15) and delta C 136.5 (C-4), 205.1 (C-3) and 171.8 (C-5) are correlated, indicating that the C-4 position is substituted with methyl; delta H 2.46 (H-6) and delta C 136.5 (C-4), 171.8 (C-5), 41.5 (C-7), 71.6 (C-8) are related, delta H 3.98 (H-8) and delta C 44.2 (C-9), 76.1 (C-10) are related, delta H 1.17 (H-14) and delta C 44.2 (C-9), 76.1 (C-10) and 83.8 (C-1) are present in relation, indicating the presence of a 5/7 bicyclic backbone with methyl substitution at position C-10; delta H 3.95 (H-13) is associated with the presence of 153.4 (C-11), 108.2 (C-12), 41.5 (C-7), indicating that C-12, C-13 and C-11 are linked and that the substituents formed are substituted at the C-7 position in combination therewith 1 H- 1 The H COSY spectrum is correlated, thus determining the planar structure of compound 11.
The relative configuration was determined by NOESY and calculated nuclear magnetism, and the presence of NOE in OH-1/H-7,H-9α/H-7,H-9β/H-14, indicated that H-7 is on the same side as OH-1 and OH-10. The relative configuration was determined to be 1S,7R,8S,10R. Absolute configurations were determined to be 1s,7r,8s,10r by comparing the calculated ECD with the measured ECD.
Daphnegenol L (12) is yellow oil (methanol) and is easy to dissolve in solvents such as methanol and DMSO.(c 0.10 MeOH) according to 1 H and 13 c NMR, deducing the formula C 15 H 22 O 4 The unsaturation was 5.
1 H NMR(600MHz,DMSO-d 6 ) In the spectrum, delta H 5.24 (1H, s, 1-OH), 4.80 (1H, s, 13-OH), 4.30 (1H, s, 10-OH), suggesting the presence of 3 active hydrogen proton signals, δ H 4.95 (1 h, d, j=1.7 hz, h-12 a), 4.82 (1 h, d, j=1.7 hz, h-12 b), suggesting the presence of a proton signal, δ, of the 1 terminal double bond H 3.94 (2H, s, H-13) suggesting the presence of 1 oxymethylene proton signal, δ H 2.79 (1 h, d, j=17.3 hz, h-2 a), 2.06 (1 h, d, j=17.3 hz, h-2 b) position 1 to methylene proton signal, δ H 1.53 (3H, s, H-15), 1.19 (3H, s, H-14) are 2 methyl proton signals. 13 C-NMR(150Hz,DMSO-d 6 ) The spectrum gives 15 carbon signals, where δ C 205.4 (C-3) the hint Structure contains 1 carbonyl group, delta C 172.1 (C-5), 156.0 (C-11), 106.2 (C-12), 136.7 (C-4) suggest that the structure contains 2 double bond groups, delta C 83.2 (C-1), 73.9 (C-10), 62.9 (C-13) suggest that the structure contains 3 oxygen sp3 hybridized carbons, delta C 47.8 (C-2), 38.9 (C-9), 37.7 (C-7), 35.5 (C-6), 29.2 (C-8), 27.6 (C-14), 8.0 (C-18) are 7 SPs 3 Hybridization of the carbon signal.
HMBC spectra 1 H- 1 H COSY Spectrum confirms that Compound 12 has 2 spin systems, terminal double bond protons δ H 4.95 (H-12 a), 4.82 (H-12 a) and carbon Signal delta C 62.9 (C-13), 37.7 (C-7) are present in relation to determine that the double bond is in position 11 and is a side chain of the 2-allylic alcohol structure, methylene proton delta H 2.79 (H-2 a), 2.06 (H-2 b) and delta C 205.3 (C-3), 172.1 (C-5), 136.7 (C-4), 83.2 (C-1), 73.9 (C-10) are correlated to determine the five-membered ring structure, δ H 2.64 (H-7) and delta C 172.1 (C-5), 38.8 (C-9) are related, delta H 1.49 (H-9) and delta C 83.2 (C-1) correlation, determination of seven-membered ring Structure, delta H 1.53 (H-15) and delta C 205.4 (C-3), 172.1 (C-5) are related, the five-membered ring has methyl substitution at position 4, delta H 1.19 (H-14) and delta C 83.2 (C-1), 38.8 (C-9) and determining that the Me-14 substitution is at the C-10 position, thereby determining the planar structure of compound 12.
The relative configuration was determined by NOSEY spectra in combination with DP4+ analysis, in which H-14 was correlated with H-9α based on the correlation of H-7 with H-9β, thus determining H-7 and Me-14 different planes, and the C-1 phase relative configuration was determined by DP4+ analysis, finally determining that the relative configuration of compound 12 was 1R,7R, and 14R. The absolute configuration of the compound was determined by comparing the calculated ECD with the measured ECD, and was 1S,7S,10S.
TABLE 6 Compounds 11 and 12 1 H (600 MHz) and 13 c (150 MHz) NMR data (DMSO-d 6 )
Daphnegenol M (13) is yellow oil (methanol) and is easily dissolved in solvents such as methanol and DMSO.(c 0.10 MeOH) according to 1 H and 13 c NMR deduces that its molecular formula is C 15 H 20 O 4 The unsaturation was calculated to be 6.
1 H NMR(600MHz,DMSO-d 6 ) In the spectrum, two methyl proton signals δ are given H 0.94 (3 h, d, j=7.3 hz, h-15), 2.22 (3 h, d, j=1.3 hz, h-14), proton signal δ at the two terminal double bonds H 5.25 (1 h, dd, j=2.0, 2.0hz, h-13 a), 5.15 (1 h, dd, j=2.0, 2.0hz, h-13 b), proton signal δ on one double bond H 5.89 (1 h, dd, j=1.3 hz, h-9) and three hydroxyl proton signals δ H 5.41(1H,d,J＝6.1Hz,OH-12),5.11(1H,s,OH-7),4.67(1H,d,J＝3.7Hz,OH-2)。 13 C NMR(150MHz,DMSO-d 6 ) A total of 15 carbon signals are shown in the spectrum, including a carbonyl carbon signal delta C 200.3 (C-8) and two sets of ethylenic carbon signals delta C 169.9 (C-10), 151.4 (C-11), 127.6 (C-9), 109.2 (C-12), 3 carbon monoxide signals delta C 81.0(C-7)72.9 (C-2), 71.0 (C-12), 1 quaternary carbon signal delta C 62.1 (C-1), 2 methine carbon signals delta C 41.6 (C-5), 31.9 (C-4), 2 methylene carbon signals delta C 41.4 (C-3), 30.2 (C-6) and 2 methyl carbon signals delta C 26.1(C-14),17.7(C-15)。
In HMBC spectra, δ H 1.85,1.59 (H-3) and delta C 72.9 (C-2), 62.1 (C-1), 41.6 (C-5), 31.9 (C-4), 26.1 (C-14) are related, indicating the presence of a five-membered ring structural fragment in the compound. Delta H 4.17 (H-12) and delta C 169.9 (C-10), 151.4 (C-11), 109.2 (C-12), 72.9 (C-2), 62.1 (C-1) are related, delta H 1.73,1.42 (H-6) and delta C 200.3 (C-8), 151.4 (C-11), 81.0 (C-7), 62.1 (C-1), 41.6 (C-5), 31.9 (C-4) are present to illustrate that the compound has a six-membered ring structural fragment, delta H 5.89 (H-9) and delta C 81.0 (C-7), 62.1 (C-1), 26.1 (C-14) are related, which indicates that a structural fragment of an alpha, beta-unsaturated ketone seven-membered ring exists, forming a 5/6/7 tricyclic skeleton. Delta H 2.22 (H-14) and delta C 169.9 (C-10), 127.6 (C-9), 62.1 (C-1) are related, delta H 0.94 (H-15) and delta C 41.4 (C-3), 31.9 (C-4), 41.6 (C-5) presence correlation determines the position of attachment of two methyl groups. And determining that the compound is the guaiane type sesquiterpene after the transformation by combining the signals, and determining the plane structure of the compound.
The relative configuration was determined by NOSEY profile in combination with dp4+ analysis, and the relative configuration of compound 13 was determined to be 1R,2S,4S,5S,7S,12S. The absolute configuration of the compound was determined by comparing the calculated ECD with the measured ECD, and the absolute configuration of the compound was 1R,2S,4S,5S,7S,12S.
TABLE 7 Compound 13 1 H (600 MHz) and 13 c (150 MHz) NMR data (DMSO-d 6 )
A pharmaceutical composition comprising said guaiane-type sesquiterpenes 1-13 and a pharmaceutically acceptable carrier.
The invention also provides application of the guaiane sesquiterpene compound 1-13 or the pharmaceutical composition containing the compound in anti-Alzheimer disease drugs.
The anti-acetylcholinesterase activities of the 13 novel guaiane sesquiterpenoids of the invention were examined, and the presence or absence of acetylcholinesterase inhibitory activity of the compounds was evaluated by taking positive control donepezil base as a reference, wherein the inhibition rate of compounds 6, 11-13 at 100. Mu.M was higher than that of the positive drug, and the inhibition rates of compounds 12 and 13 at 10. Mu.M were close to or higher than that of the positive drug. Therefore, the guaiane sesquiterpene compound has the prospect of further developing medicines for preventing and treating Alzheimer's disease.
The invention has the advantages that the compounds are novel compounds, have novel structures, are optical pure compounds with determined three-dimensional configuration, have strong anti-Alzheimer disease effect and have further development value.
Drawings
FIG. 1 Compound 1 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 2 Compound 1 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 3 Compound 2 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 4 Compound 2 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 5 Compound 3 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 6 Compound 3 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 7 Compound 4 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 8 Compound 4 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 9 Compound 5 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 10 Compound 5 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 11 is a view ofCompound 6 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 12 Compound 6 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 13 Compound 7 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 14 Compound 7 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 15 Compound 8 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 16 Compound 8 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 17 Compound 9 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 18 Compound 9 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 19 Compound 10 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 20 Compound 10 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 21 Compound 11 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 22 Compound 11 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 23 Compound 12 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 24 Compound 12 13 C-NMR spectrum (150 MHz, DMSO-d 6 )；
FIG. 25 Compound 13 1 H-NMR spectrum (600 MHz, DMSO-d) 6 )；
FIG. 26 Compound 13 13 C-NMR spectrum (150 MHz, DMSO-d 6 )。
Detailed Description
The examples set forth below are presented to aid one skilled in the art in a better understanding of the present invention and are not intended to limit the invention in any way.
Example 1
The preparation method of the guaiane sesquiterpenoids 1-13 separated from the lilac daphne root comprises the following specific operation steps:
reflux-extracting 50KG dried radix Genkwa with 70% industrial ethanol (100L each time) for 2 times and 2 hours each time, mixing the extracts, concentrating to obtain extract, extracting the extract with ethyl acetate, subjecting the obtained component to silica gel column chromatography, and performing isocratic gradient elution with dichloromethane-methanol system 100:0,90:10,80:20,70:30,60:40,50:50,40:60,30:70,20:80,10:90,0:100 (v/v), and collecting 3 fractions A-C.
Fraction B was subjected to HP20 column chromatography with a gradient of ethanol-water systems 0:100,10:90,20:80,30:70,40:60,50:50,60:40,70:30,80:20,90:10,100:0 (v/v) to give four components B1, B2, B3 and B4. Component B1 was eluted with a gradient of ethanol-water system 10:90,20:80,30:70,40:60,50:50,60:40,70:30,80:20,90:10 (v/v) by ODS column chromatography to give five components a-e.
The component b obtained was subjected to silica gel column chromatography with a petroleum ether-ethyl acetate system of 100:1,80:1,60:1,40:1,20:1,10:1 (v/v) to give four subfractions b1-b4 on the basis of TLC analysis.
Separating b1 and b2 on preparative reverse phase high performance liquid chromatography using a mobile phase of methanol-water (70:30, v/v) gives compounds 1-13, wherein the amount of the separated compounds 1-13 is 3.2mg,8.3mg,10.1mg,13.1mg,2.1mg,5.6mg,8.7mg,11.0mg,2.3mg,3.6mg,5.1mg,8.3mg,2.8mg, respectively.
Example 2
Acetylcholinesterase inhibition activity of guaiane-type sesquiterpenoids 1-13 separated from lilac daphne root is examined.
The guaiane sesquiterpenoids 1-13 prepared in example 1 were prepared in 1mg/mL stock with 1% DMSO in PBS buffer. DTNB as a substrate can be hydrolyzed by acetylcholinesterase, and the hydrolysis product can react with a color reagent ATCI to turn yellow and absorb at 412 nm. Adding a reaction reagent into a 96-well plate according to a test method, refrigerating a blank control group, a negative control group, a sample group and a sample control group overnight, adding 0.5mM ATCI, reacting for 5min, and measuring the absorbance at 412nm by using an enzyme-labeling instrument.
a Donepezil base was used as a positive control.
Claims (8)
1. The guaiane sesquiterpene compound separated from the lilac daphne root is characterized by being any one of the following structures:
2. a process for the preparation of the guaiane-type sesquiterpenoids of claim 1 comprising the steps of:
extracting dried lilac daphne root with 70% -80% industrial ethanol, mixing the extracting solutions, concentrating to obtain an extract, extracting the extract with ethyl acetate, subjecting the obtained component to silica gel column chromatography, performing isocratic gradient elution with a dichloromethane-methanol system of 100:0-0:100, and collecting three fractions A-C;
performing HP20 column chromatography on the fraction B, and performing gradient elution by using an ethanol-water system of 0:100-100:0 to obtain four components B1, B2, B3 and B4; performing gradient elution on the component B1 by using an ODS column chromatography in an ethanol-water system of 10:90-90:10 to obtain five components a-e;
subjecting the component b to silica gel column chromatography with petroleum ether-ethyl acetate system 100:1-10:1 to obtain four sub-components b1-b4 on the basis of TLC analysis;
the mobile phase of methanol-water was used to separate b1 and b2 on preparative reverse phase high performance liquid chromatography to give compounds 1-13.
3. The method for preparing a guaiane-type sesquiterpene compound according to claim 2, wherein the extraction is reflux extraction for 2-3 times, each for 2-3 hours.
4. The method for producing a guaiane-type sesquiterpene compound according to claim 2, wherein the lilac Daphne root is a root of lilac Daphne [ Daphne genkwa sieb.et Zucc ] belonging to the genus Daphne of the family daphnaceae.
5. The method for preparing the guaiane sesquiterpenes according to claim 2, wherein the volume ratio of methanol to water in the mobile phase is 70:30.
6. A pharmaceutical composition comprising a guaiane-type sesquiterpene compound according to claim 1 and a pharmaceutically acceptable carrier.
7. The use of the guaiane sesquiterpenes according to claim 1 for the preparation of a medicament against alzheimer's disease.
8. The use of the pharmaceutical composition of claim 6 for preparing an anti-alzheimer's disease medicament.
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| CN110305093A (en) * | 2019-04-11 | 2019-10-08 | 沈阳药科大学 | Guainane type sequiterpene and its preparation method and application |
| CN110305092A (en) * | 2019-04-11 | 2019-10-08 | 沈阳药科大学 | Guainane sequiterpene and its preparation and application |
| CN113248361A (en) * | 2020-02-10 | 2021-08-13 | 沈阳药科大学 | Sesquiterpenoids and medical application thereof |
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| CN110305093A (en) * | 2019-04-11 | 2019-10-08 | 沈阳药科大学 | Guainane type sequiterpene and its preparation method and application |
| CN110305092A (en) * | 2019-04-11 | 2019-10-08 | 沈阳药科大学 | Guainane sequiterpene and its preparation and application |
| CN113248361A (en) * | 2020-02-10 | 2021-08-13 | 沈阳药科大学 | Sesquiterpenoids and medical application thereof |
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| Title |
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| A new sesquiterpenoid from the shoots of Iranian Daphne mucronata Royle with selective inhibition of STAT3 and Smad3/4 cancer-related signaling pathways;Ghanadian, Mustafa等;Journal of Pharmaceutical Sciences;第28卷(第1期);253-262 * |
| Rupestonic acids B-G, NO inhibitory sesquiterpenoids from Artemisia rupestris;Chen Zhang等;Bioorganic & Medicinal Chemistry Letters;第24卷(第17期);4318-4322 * |
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