CN102250181A - Polyhydroxy steroid compounds and purpose thereof - Google Patents
Polyhydroxy steroid compounds and purpose thereof Download PDFInfo
- Publication number
- CN102250181A CN102250181A CN 201110119322 CN201110119322A CN102250181A CN 102250181 A CN102250181 A CN 102250181A CN 201110119322 CN201110119322 CN 201110119322 CN 201110119322 A CN201110119322 A CN 201110119322A CN 102250181 A CN102250181 A CN 102250181A
- Authority
- CN
- China
- Prior art keywords
- beta
- steroid
- courage steroid
- compound
- alkene
- 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.)
- Granted
Links
- -1 steroid compounds Chemical class 0.000 title claims abstract description 13
- 150000001875 compounds Chemical class 0.000 claims abstract description 61
- 229930187017 sarcsteroid Natural products 0.000 claims abstract description 34
- 241000195493 Cryptophyta Species 0.000 claims abstract description 8
- 230000000844 anti-bacterial effect Effects 0.000 claims abstract description 8
- 239000000126 substance Substances 0.000 claims abstract description 8
- 239000003814 drug Substances 0.000 claims abstract description 5
- 238000002360 preparation method Methods 0.000 claims description 25
- XEKOWRVHYACXOJ-UHFFFAOYSA-N ethyl acetate Substances CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 24
- AICOOMRHRUFYCM-ZRRPKQBOSA-N oxazine, 1 Chemical compound C([C@@H]1[C@H](C(C[C@]2(C)[C@@H]([C@H](C)N(C)C)[C@H](O)C[C@]21C)=O)CC1=CC2)C[C@H]1[C@@]1(C)[C@H]2N=C(C(C)C)OC1 AICOOMRHRUFYCM-ZRRPKQBOSA-N 0.000 claims description 16
- 229940088710 antibiotic agent Drugs 0.000 claims description 3
- 235000014653 Carica parviflora Nutrition 0.000 abstract description 15
- 241000894006 Bacteria Species 0.000 abstract description 13
- 238000012360 testing method Methods 0.000 abstract description 10
- 241001400151 Anthogorgia Species 0.000 abstract description 5
- 241000233866 Fungi Species 0.000 abstract description 5
- 241001185255 Muriceopsis flavida Species 0.000 abstract description 5
- 241000512743 Sarcophyton <anthozoan> Species 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 5
- 241000242757 Anthozoa Species 0.000 abstract description 3
- 238000000338 in vitro Methods 0.000 abstract description 3
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 96
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 62
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 50
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 42
- 238000005481 NMR spectroscopy Methods 0.000 description 30
- 238000001228 spectrum Methods 0.000 description 30
- 241000124001 Alcyonacea Species 0.000 description 26
- 238000012544 monitoring process Methods 0.000 description 25
- 230000014759 maintenance of location Effects 0.000 description 21
- 239000000843 powder Substances 0.000 description 21
- 238000004440 column chromatography Methods 0.000 description 16
- 239000000499 gel Substances 0.000 description 16
- 238000000034 method Methods 0.000 description 16
- 238000004007 reversed phase HPLC Methods 0.000 description 16
- 238000010898 silica gel chromatography Methods 0.000 description 16
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 15
- 238000005160 1H NMR spectroscopy Methods 0.000 description 15
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 15
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 15
- 229910052799 carbon Inorganic materials 0.000 description 15
- 230000008878 coupling Effects 0.000 description 15
- 238000010168 coupling process Methods 0.000 description 15
- 238000005859 coupling reaction Methods 0.000 description 15
- 229910052739 hydrogen Inorganic materials 0.000 description 15
- 239000001257 hydrogen Substances 0.000 description 15
- 238000001819 mass spectrum Methods 0.000 description 15
- 230000009897 systematic effect Effects 0.000 description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 14
- 241000243321 Cnidaria Species 0.000 description 13
- 238000000605 extraction Methods 0.000 description 13
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 12
- XILIYVSXLSWUAI-UHFFFAOYSA-N 2-(diethylamino)ethyl n'-phenylcarbamimidothioate;dihydrobromide Chemical compound Br.Br.CCN(CC)CCSC(N)=NC1=CC=CC=C1 XILIYVSXLSWUAI-UHFFFAOYSA-N 0.000 description 11
- 239000007788 liquid Substances 0.000 description 11
- 239000012046 mixed solvent Substances 0.000 description 10
- 238000010828 elution Methods 0.000 description 9
- 238000005084 2D-nuclear magnetic resonance Methods 0.000 description 8
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 8
- 241000207961 Sesamum Species 0.000 description 6
- 235000003434 Sesamum indicum Nutrition 0.000 description 6
- 229930182558 Sterol Natural products 0.000 description 6
- 239000006185 dispersion Substances 0.000 description 6
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 6
- 239000000796 flavoring agent Substances 0.000 description 6
- 235000019634 flavors Nutrition 0.000 description 6
- 235000013372 meat Nutrition 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 150000003432 sterols Chemical class 0.000 description 6
- 235000003702 sterols Nutrition 0.000 description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 5
- 241001299682 Salix purpurea Species 0.000 description 5
- 238000002329 infrared spectrum Methods 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- 241000194107 Bacillus megaterium Species 0.000 description 3
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 3
- 241000242752 Octocorallia Species 0.000 description 3
- 229930182555 Penicillin Natural products 0.000 description 3
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 3
- 241001674391 Sphaerulina musiva Species 0.000 description 3
- 229960003328 benzoyl peroxide Drugs 0.000 description 3
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 3
- 229930014626 natural product Natural products 0.000 description 3
- 229940049954 penicillin Drugs 0.000 description 3
- XWBSJQAHXPZQLU-XUJKSOOQSA-N sarcsteroid C Natural products CC(C)[C@@H](C)C(=C[C@@H](C)[C@H]1CC[C@H]2[C@@H]3C[C@@H](O)[C@@]4(O)C[C@@H](O)CC[C@]4(C)[C@H]3[C@H](O)C[C@]12C)C XWBSJQAHXPZQLU-XUJKSOOQSA-N 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000003643 water by type Substances 0.000 description 3
- 241000897681 Cliona copiosa Species 0.000 description 2
- 241000700108 Ctenophora <comb jellyfish phylum> Species 0.000 description 2
- 229930004069 diterpene Natural products 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 238000013332 literature search Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 229930000044 secondary metabolite Natural products 0.000 description 2
- 150000003431 steroids Chemical class 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- QTENRWWVYAAPBI-YCRXJPFRSA-N streptomycin sulfate Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O QTENRWWVYAAPBI-YCRXJPFRSA-N 0.000 description 2
- YUZXLQMUJSICKF-UHFFFAOYSA-N (3beta,5alpha,6beta,22R,23R,24R)-23-Norgorgost-7-ene-3,5,6-triol Natural products CC(C)C(C)C1CC1C(C)C1C2(C)CCC3C4(C)CCC(O)CC4(O)C(O)C=C3C2CC1 YUZXLQMUJSICKF-UHFFFAOYSA-N 0.000 description 1
- KVUXYQHEESDGIJ-UHFFFAOYSA-N 10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthrene-3,16-diol Chemical class C1CC2CC(O)CCC2(C)C2C1C1CC(O)CC1(C)CC2 KVUXYQHEESDGIJ-UHFFFAOYSA-N 0.000 description 1
- 241000178555 Acanthogorgiidae Species 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000221495 Microbotryum violaceum Species 0.000 description 1
- 241001184767 Muriceopsis Species 0.000 description 1
- 241001223361 Sarcophyton glaucum Species 0.000 description 1
- 240000008866 Ziziphus nummularia Species 0.000 description 1
- 241001360088 Zymoseptoria tritici Species 0.000 description 1
- 241000195647 [Chlorella] fusca Species 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 150000001335 aliphatic alkanes Chemical class 0.000 description 1
- XRWSZZJLZRKHHD-WVWIJVSJSA-N asunaprevir Chemical compound O=C([C@@H]1C[C@H](CN1C(=O)[C@@H](NC(=O)OC(C)(C)C)C(C)(C)C)OC1=NC=C(C2=CC=C(Cl)C=C21)OC)N[C@]1(C(=O)NS(=O)(=O)C2CC2)C[C@H]1C=C XRWSZZJLZRKHHD-WVWIJVSJSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 229940125961 compound 24 Drugs 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000006471 dimerization reaction Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003777 experimental drug Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 1
- 238000003929 heteronuclear multiple quantum coherence Methods 0.000 description 1
- 230000033444 hydroxylation Effects 0.000 description 1
- 238000005805 hydroxylation reaction Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 150000002611 lead compounds Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 239000003186 pharmaceutical solution Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 150000003535 tetraterpenes Chemical class 0.000 description 1
- 235000009657 tetraterpenes Nutrition 0.000 description 1
Landscapes
- Steroid Compounds (AREA)
Abstract
The invention relates to the technical field of medicine. The invention provides a class of polyhydroxy steroid compounds which are separated from three corals (Muriceopsis flavida, Sarcophyton sp., and Anthogorgia sp.) collected from Hainan and have antibacterial and anti-microalgae activity, and the chemical structure formula of the polyhydroxy steroid compounds is as shown in the following picture. In vitro antibacterial tests show that the compounds, which contains Muristeroids A-G, Sarcsteroids A-F, Anthsteroids A-B, cholesteric-5alpha, 6beta-diol-3beta-acetic acid ester, cholesteric-5alpha-methoxy-3beta, 6beta-diol, (22E)-22-alkene-cholesteric-3beta, 5alpha, 6beta-triol, 24(28)-alkene-ergosta-3beta, 5alpha, 6beta-triol, cholesteric-3beta, 5alpha, 6beta-triol, cholesteric-1beta, 3beta, 5alpha and 6beta-tetrol, have an obvious effect of inhibiting fungi, bacteria and algae, and can be used for preparing antibacterial and anti-microalgae medicaments. The polyhydroxy steroid compounds provided by the invention are of great significance for developing and utilizing China's marine medicinal biological resources.
Description
Technical field
The present invention relates to medical technical field, be specifically related to from marine invertebrate-coral (Muriceopsis flavida, Sarcophyton sp., Anthogorgia sp.) separate in 15 of obtaining new have antibiotic, the poly-hydroxy steroid compound Muristeroids A～G of anti-activity of microalgae, Sarcsteroids A～F, AnthsteroidsA～B and 6 are known have antibiotic, the poly-hydroxy steroid compound of anti-activity of microalgae: courage steroid-5 α, 6 beta-diols-3 β-acetic ester, courage steroid-5 α-methoxyl group-3 β, 6 beta-diols, (22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α, 6 beta-triols, courage steroid-1 β, 3 β, 5 α, the new purposes of 6 β-tetrol.
Background technology
South China Sea located in subtropical zone zone, the coral resource is particularly abundant, is coral one of the marine site of concentrated distribution in the world, and in more than the 6100 kind of coral that the whole world is found, Chinese marine site just has 719 kinds at present.Coral is no vertebra lower animal, for numerous biologies provides the habitat, and as crinosity section, bivalve section, abdominal foot section, fish etc.Though coral self lacks effective physics defence capability, can secrete some chemical defence materials, resist the invasion of protozoon and other biological, this phenomenon is to study the theoretical basis of secondary metabolite in the corallum in the world.Coral can be divided into alcyon and two subclasses of zoantharian according to the number of branches of gastrovascular cavity, and soft coral in the Alcyonaria and gorgonian become the focus of marine natural product research because of containing novel structure, the good secondary metabolite of biological activity.
Purple willow coral (Muriceopsis flavida) belongs to Coelenterata (Coelenterata) Anthozoa (Anthoaoa) Alcyonaria (Octocorallia) Gorgonacea (Gorgonacea) the clump Muriceopsis of gorgonian section (Pexuridae) and belongs to animal.The purple willow coral mainly is distributed in east, the Pacific Ocean, California seashore and Caribbean Division.At present, the report of relevant this genus coral chemical constitution study has only one piece both at home and abroad, 5 four sterol that methylates [Kokke WCMC have therefrom been obtained, Bihlin L, Fenical W, Djerassi C.Novel dinoflagaellate 4 α-methylated sterols from four Caribbean gorgonians[J] .Phytochemistry, 1982,21 (4): 881-887], do not see the pertinent literature of its active report.
Meat sesame soft coral (Sarcophyton sp.) belongs to soft coral order (Alcyonacea) soft coral section (Alcyonacea) meat sesame soft coral and belongs to (Sarcophyton) animal.It is various that this belongs to the soft coral kind, and it is wide to distribute.Highly oxidized lax alkane diterpene and sterol are the main metabolites of this genus soft coral, except lax alkane diterpene and sterol, have also therefrom found the tetraterpene of the lax alkane type of dimerization that structure is very peculiar.More than 180 new compounds from this genus coral, have been found so far.India scholar Anjaneyulu had once done detailed summary [Anjaneuyulu A.S.R., Rao G.V.J.Ind.Chem.Soc., 1997,74,272.] to the chemical constitution study situation of meat sesame soft coral.
Piece flower gorgonian (Anthogorgia sp.) belongs to Gorgonacea (Gorginacea) sour jujube gorgonian section (Acanthogorgiidae) animal.Literature search finds no the relevant report of closing piece flower gorgonian chemical constitution study.
Summary of the invention
The invention provides 21 poly-hydroxy steroid compounds that extraction separation obtains from three kinds of South Sea corals (Muriceopsis flavida, Sarcophyton sp., Anthogorgia sp.), difference called after Muristeroids A～G, Sarcsteroid A～F, Anthsteroids A～B, courage steroid-5 α, 6 beta-diols-3 β-acetic ester, courage steroid-5 α-methoxyl group-3 β, 6 beta-diols, (22E)-and 22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α, 6 beta-triols, courage steroid-1 β, 3 β, 5 α, 6 β-tetrol.Wherein compound Muristeroids A～G and courage steroid-5 α, 6 beta-diols-3 β-acetic ester, courage steroid-5 α-methoxyl group-3 β, 6 beta-diols, (22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α, 6 beta-triols, courage steroid-1 β, 3 β, 5 α, 6 β-tetrol separates from the purple willow coral and obtains, compound S arcsteroids A～F separates from meat sesame soft coral and obtains, and compd A nthsteroids A～B separates from piece flower gorgonian and obtains, and their chemical structure of general formula is as follows:
Wherein Muristeroids A～G, Sarcsteroids A～F, Anthsteroids A～B are new compound.
Muristeroid A is a white powder, and systematic naming method falls courage steroid-5 α for (22E)-22-alkene-24-, 6 beta-diols-3 β-acetic ester.The accurate molecular weight m/z 469.3294[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of carbon spectrum hydrogen spectrum, the molecular formula that provides this compound is C
28H
46O
4, molecular weight is 446;
IR (film) v
Max: 3454,2953,2941,2867,1716,1599,1496,1452,1382,1367,1281,1161,1037,968,699; UV (CHCl
3) λ
MaxNm (Abs): 257 (0.3642);
1H and
13The C nuclear magnetic resonance data sees Table 1.
Muristeroid B is a white powder, and systematic naming method is (22E)-22-alkene-courage steroid-5 α, 6 beta-diols-3 β-acetic ester.The accurate molecular weight m/z483.2188[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, reach carbon spectrum hydrogen spectrum and resolve, provide the molecular formula C of this compound
29H
48O
4, molecular weight 460;
Infrared spectra IR (film) v
Max: 3369,2956,2927,1736,1664,1599,1458,1378,1260,1043,968,699; UV (CHCl
3) λ
MaxNm (Abs): 240 (0.8793);
1H and
13The C nuclear magnetic resonance data sees Table 2.
Muristeroid C is a white powder, and systematic naming method is (22E)-22-alkene-5 α-methoxyl group-courage steroid-3 β, 6 beta-diols.The accurate molecular weight m/z455.3498[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
28H
48O
3, molecular weight 432;
Infrared spectra IR (film) v
Max: 3374,3026,2929,2867,1600,1496,1454,1379,1262,1078,1032,754,699; UV (CHCl
3) λ
MaxNm (Abs): 337 (0.4653);
1H and
13The C nuclear magnetic resonance data sees Table 3.
Muristeroid D is a white powder, and systematic naming method falls courage steroid-3 β for (22E)-22-alkene-24-, 5 α, 6 beta-diols.The accurate molecular weight m/z427.6338[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
26H
44O
3, molecular weight 404;
Infrared spectra IR (film) v
Max: 3373,3026,2959,2928,2854,1600,1496,1453,1379,1302,1260,1040,966,749,699; UV (CHCl
3) λ
MaxNm (Abs): 278 (0.4411);
1H and
13The C nuclear magnetic resonance data sees Table 4.
Muristeroid E is a white powder, and systematic naming method is 24 (25)-alkene-courage steroid-3 β, 5 α, 6 beta-triols.The accurate molecular weight m/z 441.6371[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, to resolve in conjunction with hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
27H
46O
3, molecular weight 418; ,
1H-NMR,
13C-NMR, DEPT,
1H-
1HCOSY, NOESY, HMBC, HMQC determine compound structure,
1H and
13The C nuclear magnetic resonance data sees Table 5.
Muristeroid F is a white powder, and systematic naming method falls courage steroid 1 β for (22E)-22-alkene-24-, 3 β, 5 α, 6 β-tetrol.The accurate molecular weight m/z443.2857[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
26H
44O
4, molecular weight 420;
Infrared spectra IR (film) v
Max: 3316,2953,2926,2851,1600,1496,1455,1379,1212,1098,1040,1009,960; UV (CHCl
3) λ
MaxNm (Abs): 278 (0.1204);
1H and
13The C nuclear magnetic resonance data sees Table 6.
Muristeroid G is a white powder, and systematic naming method is (22E)-22-alkene-courage steroid 1 β, 3 β, 5 α, 6 β-tetrol.The accurate molecular weight m/z 457.3291[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
26H
44O
4, molecular weight 434;
Infrared spectra IR (film) v
Max: 3337,2951,2927,2867,1604,1459,1371,1042,1007,964;
1H and
13The C nuclear magnetic resonance data sees Table 7.
Courage steroid-5 α, 6 beta-diols-3 β-acetic ester: be white powder, by ESI-MS,
1H-NMR,
13C-NMR, DEPT and determine compound structure [Joao FS Carvalho, M Manuel Cruz Silva, MLuisa Sa Emelo.Efficient trans-diaxial hydroxylation of △ in conjunction with document
5Steroids[J] .Tetrahedron, 2010,66; 2455-2462].
Courage steroid-5 α-methoxyl group-3 β, 6 beta-diols: be white powder.By ESI-MS,
1H-NMR,
13C-NMR, DEPT and determine compound structure [Blunt J W in conjunction with document, Fischer A, Hartshorn M P, Jones FW, Kirk D N, Yoong S W.Acid catalyzed reactions of 5 α-hydroxy steroids.III.Westphalen rearrangement[J] .Tetrahedron, 1965,21 (6); 1567-1580].
(22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols: white powder, by ESIMS,
1H-NMR,
13C-NMR also determines compound structure [Giacino Notaro in conjunction with document, Bincenzo Piccialli, Donato Sica.3 β, 5 α, 6 β-trihydrolylated sterols with a saturated mucleus from two populations of the marine sponge Cliona copiosa[J] .Journal of natural products, 1991,54 (6); 1570-1575].
24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols: white powder, by ESIMS,
1H-NMR,
13C-NMR also determines compound structure [Kobasyashi in conjunction with document, Masaru, Madala M Krishna, et al.Isolation of 23-Demethylgorgost-7-ene-3 β, 5 α, 6 β-triol and (24)-Ergostane-3 β, 5 α, 6 β, 7 β, 15 β-pentol from soft corals of the Andaman and Micobar coasts[J] .Chemical ﹠amp; Phamaceutical Bulletin, 1993,41 (1); 87-89].
Courage steroid-3 β, 5c, 6 beta-triols: white powder, by ESIMS,
1H-NMR,
13C-NMR also determines compound structure [Giacomo Notaro in conjunction with document, Vincenzo Piccialli, Donato Sica.3 β, 5 α, 6 β-trihydrolylated sterols with a saturated nucleus from two populations of the marine sponge Cliona copiosa[J] .Journal of natural products.1991,54 (6); 1570-1575].
Courage steroid-1 β, 3 β, 5 α, 6 β-tetrol; White powder, by ESIMS,
1H-NMR,
13C-NMR also determines compound structure [Kobayashi Masaru in conjunction with document, Hayashi Takaaki, Nakajima Fumie, Mitsuhashi Hiroshi.Marine steroids.IX.Occurrence of 24 ε-methylcholastane-1 β, 3 β, 5 α, 6 β, 25-pentol-25-monoacetate in the soft coral Sarcophyton glaucum[J] .Steroids, 1979,34 (3); 285-293].
Sarcsteroid A is a white powder, and systematic naming method is 11 α-acetoxyl group-24-methyl-22 (23), 25 (27)-diene-courage steroid-3 β, 5 α, 6 beta-triols.The accurate molecular weight m/z 511.2634[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
30H
48O
5, molecular weight 488; By
1H-NMR,
13C-NMR, DEPT, and a complete set of two dimensional NMR is determined compound structure;
1H and
13The C nuclear magnetic resonance data sees Table 8.
Sarcsteroid B is white amorphous powder, and systematic naming method is 11 α-acetoxyl group-24 (25)-alkene-courage steroid-3 β, 5 α, 6 beta-triols.The accurate molecular weight m/z 499.4274[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
29H
48O
5, molecular weight 476; By
1H-NMR,
13C-NMR, DEPT, and a complete set of two dimensional NMR is determined compound structure;
1H and
13The C nuclear magnetic resonance data sees Table 9.
Sarcsteroid C is the white powder solid, and systematic naming method is 23,24-dimethyl-22 (23)-diene-courage steroid-3 β, 5 α, 6 beta-triols.The accurate molecular weight m/z 485.3532[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
29H
50O
5, molecular weight 478; By
1H-NMR,
13C-NMR, DEPT, and a complete set of two dimensional NMR is determined compound structure;
1H and
13The C nuclear magnetic resonance data sees Table 10.
Sarcsteroid D is the white powder solid, and systematic naming method is 23,24-dimethyl-22 (23)-diene-courage steroid-3 β, 5 α, 6 beta-triols.The accurate molecular weight m/z 485.3532[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
29H
50O
5, molecular weight 478; By
1H-NMR,
13C-NMR, DEPT, and a complete set of two dimensional NMR is determined compound structure;
1H and
13The C nuclear magnetic resonance data sees Table 11.
Sarcsteroid E is the white powder solid, and systematic naming method is 11 α-acetoxyl group-gorgostane-3 β, 5 α, 6 β, 12 α-tetrol.The accurate molecular weight m/z 557.5432[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
32H
54O
6, molecular weight 534; By
1H-NMR,
13C-NMR, DEPT, and a complete set of two dimensional NMR is determined compound structure;
1H and
13The C nuclear magnetic resonance data sees Table 12.
Sarcsteroid F is the white powder solid, and systematic naming method is (24S)-24-methyl-courage steroid-1 α, 3 β, 5 α, 6 β, 11 α-pentol.The accurate molecular weight m/z 489.4582[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
28H
50O
5, molecular weight 466; By
1H-NMR,
13C-NMR, DEPT, and a complete set of two dimensional NMR is determined compound structure;
1H and
13The C nuclear magnetic resonance data sees Table 13.
AnthsteroidA is the white powder solid, and systematic naming method is courage steroid-3 β, 5 α, 6 β, 11 α-tetrol.The accurate molecular weight m/z 435.3476[M-H that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
-, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
27H
48O
4, molecular weight 436; By
1H-NMR,
13C-NMR, DEPT, and a complete set of two dimensional NMR is determined compound structure;
1H and
13The C nuclear magnetic resonance data sees Table 14.
Anthsteroid B is the white powder solid, and systematic naming method is 22 (23)-alkene-courage steroid-3 β, 5 α, 6 β, 11 α-tetrol.The accurate molecular weight m/z 457.3295[M+Na that provides by quasi-molecular ion peak in the high resolution mass spectrum (HR-ESI-MS)]
+, and the parsing of hydrogen spectrum carbon spectrum, the molecular formula that provides this compound is C
27H
46O
4, molecular weight 434; By
1H-NMR,
13C-NMR, DEPT, and a complete set of two dimensional NMR is determined compound structure;
1H and
13The C nuclear magnetic resonance data sees Table 15.
Table 1.Muristeroid A's
1H and
13C nuclear magnetic resonance data (CDCl
3, 400MHz/100MHz)
J value: coupling constant
Table 2.Muristeroid B's
1H and
13C nuclear magnetic resonance data (CDCl
3, 400MHz/100MHz)
J value: coupling constant
Table 3.Muristeroid C's
1H and
13C nuclear magnetic resonance data (CDCl
3, 400MHz/100MHz)
J value: coupling constant
Table 4.Muristeroid D's
1H and
13C nuclear magnetic resonance data (CD
30D, 400MHz/100MHz)
J value: coupling constant
Table 5.Muristeroid E's
1H and
13C nuclear magnetic resonance data (CDCl
3, 500MHz/100MHz)
J value: coupling constant
Table 6.Muristeroid F's
1H and
13C nuclear magnetic resonance data (CD
3OD, 400MHz/100MHz)
J value: coupling constant
Table 7.Muristeroid G's
1H and
13C nuclear magnetic resonance data (CD
3OD, 400MHz/100MHz)
J value: coupling constant
Table 8.Sarcsteroid A's
1H and
13C nuclear magnetic resonance data (CDCl
3, 400MHz/100MHz)
J value: coupling constant
Table 9.Sarcsteroid B's
1H and
13C nuclear magnetic resonance data (CDCl
3, 400MHz/100MHz)
J value: coupling constant
Table 10.Sarcsteroid C's
1H and
13C nuclear magnetic resonance data (CDCl
3, 400MHz/100MHz)
J value: coupling constant
Table 11.Sarcsteroid D's
1H and
13C nuclear magnetic resonance data (CDCl
3, 400MHz/100MHz)
J value: coupling constant
Table 12.Sarcsteroid E's
1H and
13C nuclear magnetic resonance data (CDCl
3, 400MHz/100MHz)
J value: coupling constant
Table 13.Sarcsteroid F's
1H and
13C nuclear magnetic resonance data (CDCl
3, 500MHz/100MHz)
J value: coupling constant
Table 14.Anthsteroid A's
1H and
13C nuclear magnetic resonance data (CD
3OD, 400MHz/100MHz)
J value: coupling constant
Table 15.Anthsteroid B's
1H and
13C nuclear magnetic resonance data (CD
3OD, 400MHz/100MHz)
J value: coupling constant
Muristeroids A～G of the present invention, courage steroid-5 α, 6 beta-diols-3 β-acetic ester, courage steroid-5 α-methoxyl group-3 β, 6 beta-diols, (22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α, 6 beta-triols, courage steroid-1 β, 3 β, 5 α, 6 β-tetrol, Sarcsteroids A～F, the preparation method of AnthsteroidsA～B is as follows:
1.Muristeroids the preparation of A～G and 6 known compounds
Get fresh purple willow coral and clean and to shred, with weight ratio be 5～10 times acetone (adding less water in the acetone) supersound extraction to extracting liquid colourless, reclaim under reduced pressure acetone is distinguished the flavor of to there being acetone, use the equal-volume water-dispersion, extracted with diethyl ether several, combining extraction liquid, the reclaim under reduced pressure ether gets total medicinal extract to doing.Total medicinal extract is separated with normal phase silica gel chromatography (200～300 order),,, collect, total medicinal extract is merged into 16 parts according to cut polarity size by the thin layer plate monitoring with sherwood oil/acetone system gradient elution.
Fr.6 partly pass through Sephandex LH-20 gel column chromatography (moving phase: sherwood oil: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.6-1～Fr.6-6.Fr.6-5 part is passed through normal phase silica gel chromatography (400-600 order) again and separated sherwood oil: 4: 1 wash-outs of acetone, the thin layer plate monitoring is divided into 4 parts (Fr.6-5-1～Fr.6-5-4) with component.Fr.6-5-2 is by half preparation type RP-HPLC purifying, with methyl alcohol: the volume ratio of water is that 90: 10 mixed solvent carries out wash-out, the differential detector monitors, collection contains Muristeroid C and courage steroid-5 α-methoxyl group-3 β, stream part of 6 beta-diols, reclaim under reduced pressure gets compound Muristeroid C and courage steroid-5 α-methoxyl group-3 β, 6 beta-diols to doing.Fr.6-5-3 is by half preparation type RP-HPLC purifying, with methyl alcohol: the volume ratio of water is that 95: 5 mixed solvent carries out wash-out, the differential detector monitors, collection contains Muristeroid A, B and courage steroid-5 α, stream part of 6 beta-diols-3 β-acetic ester, reclaim under reduced pressure gets compound MuristeroidA, Muristeroid B and courage steroid-5 α, 6 beta-diols-3 β-acetic ester to doing.
Add The addition of C HCl in the Fr.12 part
3, filter, get filter residue, with filter residue CH
3OH: CHCl
31: 1 mixed solvent dissolving separates (moving phase CH through Sephadex LH-20 gel column chromatography
3OH: CHCl
3=1: 1), thin layer plate detects, collect merge Fr.12-1～Fr.12-4, Fr.12-3 is with half preparation type RP-HPLC purifying (moving phase, 85% methanol-water), the parallax detector detects, and collects to contain MuristeroidsD, Muristeroids E, (22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α, stream part of 6 beta-triols, reclaim under reduced pressure gets compound Muristeroids D to doing, Muristeroids E, (22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α t, 6 beta-triols.
Fr.13 part is separated through normal phase silica gel column chromatography, by the chloroform/methanol gradient elution, detects monitoring according to thin layer plate, collect merge Fr.13-1～Fr.13-4.Add an amount of chloroform among the Fr.13-3, filter, get filter residue.With the chloroform/methanol dissolving of filter residue, separate (moving phase, CH through Sephadex LH-20 gel column chromatography with 1: 1
3OH: CHCl
3=1: 1).Thin layer plate detects, collect merge Fr.13-3-1～Fr.13-3-3.Fr.13-3-2 is with half preparation type RP-HPLC purifying (moving phase, 85% methanol-water), the parallax detector detects, and collects to contain Muristeroids F～G, courage steroid-1 β, 3 β, 5 α, stream part of 6 β-tetrol, reclaim under reduced pressure is to doing, get compound Muristeroid F, Muristeroid G and courage steroid-1 β, 3 β, 5 α, 6 β-tetrol.
2.Sarcsteroids the preparation of A～F
Get green meat sesame soft coral and clean and to shred, with weight ratio be 5～10 times acetone (adding less water in the acetone) supersound extraction to extracting liquid colourless, reclaim under reduced pressure acetone is distinguished the flavor of to there being acetone, use the equal-volume water-dispersion, extracted with diethyl ether several, combining extraction liquid, the reclaim under reduced pressure ether gets total medicinal extract to doing.Total medicinal extract is separated with normal phase silica gel chromatography (200～300 order),,, collect, total medicinal extract is merged into 23 parts according to cut polarity size by the thin layer plate monitoring with n-hexane/acetone system gradient elution.
Fr.20 partly pass through Sephandex LH-20 gel column chromatography (moving phase: normal hexane: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.20-1～Fr.20-7.Fr.20-3 part pass through again Sephandex LH-20 gel column chromatography (moving phase: normal hexane: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.20-3-1～Fr.20-3-4.Fr.20-3-3 is by half preparation type RP-HPLC purifying, with methyl alcohol: the volume ratio of water is that 96: 4 mixed solvent carries out wash-out, the differential detector monitors, collection contains stream part of Sarcsteroid A, Sarcsteroid B, Sarcsteroid D, reclaim under reduced pressure gets compound S acrsteroid A, Sacrsteroid B, Sacrsteroid D to doing.
Fr.21 partly pass through Sephandex LH-20 gel column chromatography (moving phase: normal hexane: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.21-1～Fr.21-7.Fr.21-5 part pass through again normal phase silica gel column chromatography (moving phase: methyl alcohol: wash-out chloroform=1: 50), detect monitoring according to thin layer plate, collect merge Fr.21-5-1～Fr.21-5-5.Fr.21-5-5 is by half preparation type RP-HPLC purifying, with methyl alcohol: the volume ratio of water is that 95: 5 mixed solvent carries out wash-out, the differential detector monitors, collection contains stream part of Sarcsteroid E, Sarcsteroid F, reclaim under reduced pressure gets compound S arcsteroidE, Sarcsteroid F to doing.
Fr.22 partly pass through Sephandex LH-20 gel column chromatography (moving phase: normal hexane: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.22-1～Fr.22-6.Fr.22-3 part pass through again Sephandex LH-20 gel column chromatography (moving phase: normal hexane: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.22-3-1～Fr.22-3-6.Fr.22-3-5 is by normal phase silica gel column chromatography (moving phase: chloroform/methanol=40: 1) wash-out, again through more than half preparation type RP-HPLC purifying, with methyl alcohol: the volume ratio of water is that 94: 6 mixed solvent carries out wash-out, the differential detector monitors, collection contains stream part of Sarcsteroid C, reclaim under reduced pressure gets compound S arcsteroid C to doing.
3.Anthsteroids the preparation of A～B
Get fresh block of colored gorgonian and clean and to shred, with weight ratio be 5～10 times acetone (adding less water in the acetone) supersound extraction to extracting liquid colourless, reclaim under reduced pressure acetone is distinguished the flavor of to there being acetone, use the equal-volume water-dispersion, extracted with diethyl ether several, combining extraction liquid, the reclaim under reduced pressure ether gets total medicinal extract to doing.Total medicinal extract is separated with normal phase silica gel chromatography (200～300 order),,, collect, total medicinal extract is merged into 32 parts according to cut polarity size by the thin layer plate monitoring with sherwood oil/acetone system gradient elution.
Fr.21 partly passes through reversed-phase silica gel column chromatography (moving phase: methyl alcohol: water=50%, 70%, 80%, 90%) wash-out, detects monitoring according to thin layer plate, collect merge Fr.21-1～Fr.21-6.Fr.21-5 is by half preparation type RP-HPLC purifying, with methyl alcohol: the volume ratio of water is that 75: 25 mixed solvent carries out wash-out, the differential detector monitors, collection contains stream part of Anthsteroid A and Anthsteroid B, reclaim under reduced pressure gets compd A nthsteroid A and Anthsteroid B to doing.
Literature search shows, compound Muristeroids A～G, courage steroid-5 α, 6 beta-diols-3 β-acetic ester, courage steroid-5 α-methoxyl group-3 β, 6 beta-diols, (22E)-and 22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α, 6 beta-triols, courage steroid-1 β, 3 β, 5 α, 6 β-tetrol, Sarcsteroids A～F, Anthsteroids A～B all do not have the research report of antibiotic and activity of microalgae.
The present invention shows in-vitro antibacterial and the anti-little algae test that above-mentioned 21 compounds carry out, they have the obvious suppression effect for test with fungi, bacterium and algae, therefore, the invention provides the application of above-mentioned poly-hydroxy steroid compound in preparing antibiotic, anti-little algae medicine.
Embodiment
Now in conjunction with the embodiments the present invention is described in detail.
Embodiment 1. preparation Muristeroids A～G and 6 known compounds.
Get purple willow coral (Muriceopsis flavida) 1500g of surrounding waters, Sanya, Chinese Hainan, clean, shred, with the acetone supersound extraction of 5 times of weight, reclaim under reduced pressure acetone is to there not being the acetone flavor, use the equal-volume water-dispersion, extracted with diethyl ether 6 times, each 1000ml, combining extraction liquid, reclaim ether and be concentrated into dried, total medicinal extract 12.5g.Total medicinal extract is separated through normal phase silica gel chromatography (200～300 order), it with volume ratio 80: 1,40: 1,20: 1,10: 1,5: 1,3: 1,2: 1,1: 1,1: 5,1: 15 petrol ether/ethyl acetate gradient elution, monitor according to thin layer plate, each flow point is collected by polarity size, collect altogether 16 part Fr.1～Fr.16.
Fr.6 partly use Sephandex LH-20 gel column chromatography (mobile sherwood oil: methyl alcohol: chloroform=2: 1: 1) separate,, collect and merge stream part according to the thin layer plate monitoring, six part Fr.6-1～Fr.6-6.Fr.6-5 part is passed through normal phase silica gel chromatography (400-600 order) separation and purification (4: 1 wash-outs of sherwood oil/acetone) again, and the thin layer plate monitoring is divided into 4 parts (Fr.6-5-1～Fr.6-5-4) with component.Fr.6-5-2 is by partly preparing the RP-HPLC purifying, with methyl alcohol: the volume ratio of water is that 90: 10 mixed solvent is a moving phase, differential detector monitors, control flow velocity 1.5ml/min, 30 ℃ of constant column temperature temperature, stream part when the collection retention time is 30min, reclaim under reduced pressure gets compound Muristeroid C (1.3mg) to doing, stream part when the collection retention time is 45min, reclaim under reduced pressure gets compound courage steroid-5 α-methoxyl group-3 β, 6 beta-diols (1.6mg) to doing.Fr.6-5-3 is by half preparation type RP-HPLC purifying, with methyl alcohol: the volume ratio of water is that 95: 5 mixed solvent is a moving phase, the differential detector monitors, control flow velocity 1.5ml/min, 30 ℃ of constant column temperature temperature, stream part when the collection retention time is 30min, reclaim under reduced pressure gets compound MuristeroidA (8mg) to doing, the stream part when the collection retention time is 42min, reclaim under reduced pressure is to doing, get compound Muristeroid B (5mg), the stream part when the collection retention time is 55min, reclaim under reduced pressure is to doing, get compound courage steroid-5 α, 6 beta-diols-3 β-acetic ester (3mg).
Add an amount of chloroform in the Fr.12 part, filter, get filter residue, use chloroform: the dissolving of 1: 1 mixed solvent of methyl alcohol, with Sephandex LH-20 gel column chromatography (mobile methyl alcohol: chloroform=1: 1) separate, monitor according to thin layer plate, collect to merge and flow part, get four part Fr.12-1～Fr.12-4.The Fr.12-3 part is again through more than half preparation RP-HPLC purifying (methanol-waters of moving phase 85%; Flow velocity: 1.5ml/min; Column temperature: 30 ℃), differential detector monitors, the stream part when the collection retention time is 52min, reclaim under reduced pressure gets compound Muristeroid D (2mg) to doing, the stream part when the collection retention time is 68min, reclaim under reduced pressure gets compound Muristeroid E (2mg) to doing; Stream part when the collection retention time is 75min, reclaim under reduced pressure gets compound (22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols (5.5mg) to doing; Stream part when the collection retention time is 90min, reclaim under reduced pressure gets compound 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols (3.5mg) to doing; Stream part when the collection retention time is 119min, reclaim under reduced pressure gets compound courage steroid-3 β, 5 α, 6 beta-triols (11.0mg) to doing.
Fr.13 part is separated through normal phase silica gel column chromatography, by chloroform: methyl alcohol 40: 1,30: 1,20: 1,15: 1, for eluent do gradient elution at 10: 1, detect monitoring according to thin layer plate, collect and merge stream part, four part Fr.13-1～Fr.13-4.Add an amount of chloroform among the Fr.13-3, filter, get filter residue, with the chloroform of filter residue with 1: 1: dissolve with methanol, through Sephadex LH-20 gel column chromatography separation (moving phase CH
3OH: CHCl
3=1: 1).The thin layer plate detection is monitored, and collects to merge stream part, gets three part Fr.13-3-1～Fr.13-3-3.Fr.13-3-2 is with the half preparation type RP-HPLC purifying (methanol-water of moving phase 85%; Flow velocity: 1.5ml/min; Column temperature: 30 ℃), the parallax detector monitors is collected the stream part when containing retention time 66min, and reclaim under reduced pressure gets compound Muristeroid F (2.5mg) to doing; Stream part when collecting retention time 78min, reclaim under reduced pressure gets compound Muristeroid G (5.0mg) to doing; Stream part when collecting retention time 85min, reclaim under reduced pressure gets compound courage steroid-1 β, 3 β, 5 α, 6 β-tetrol (8.0mg) to doing.
Embodiment 2. preparation Sarcsteroids A～F
Get meat sesame soft coral (Sarcophytum sp) 1642g of surrounding waters, the Guangxi China North Sea, clean, shred, with the acetone supersound extraction of 5 times of weight, reclaim under reduced pressure acetone is to there not being the acetone flavor, use the equal-volume water-dispersion, extracted with diethyl ether 6 times, each 1000ml, combining extraction liquid, reclaim ether and be concentrated into dried, total medicinal extract 21.6g.Total medicinal extract is separated through normal phase silica gel chromatography (200～300 order), with volume ratio is that gradient elution is done in 99: 1,79: 1,69: 1,59: 1,49: 1,39: 1,29: 1,25: 1,22: 1,17: 1,15: 1,12: 1,9: 1,8: 1,7: 1,6: 1,5: 1,4: 1,3: 1,2: 1,1: 1,1: 2,1: 5,1: 10, the petrol ether/ethyl acetate of 100% ethyl acetate, according to the thin layer plate monitoring, each flow point is collected altogether from small to large by the polarity size and is merged into 23 part Fr.1～Fr.23.
Fr.20 (210mg) part through Sephandex LH-20 gel column chromatography (moving phase: normal hexane: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.20-1～Fr.20-7.Fr.20-3 (72mg) part pass through again Sephandex LH-20 gel column chromatography (moving phase normal hexane: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.20-3-1～Fr.20-3-4.Fr.20-3-3 is by the half preparation type RP-HPLC purifying (methanol-water of moving phase 96%; Flow velocity 1.5ml/min; 30 ℃ of column temperatures), the differential detector monitors, stream part when collection contains retention time and is 19min, reclaim under reduced pressure is to doing, get Sarcsteroid A (2.3mg), the stream part when the collection retention time is 26min, reclaim under reduced pressure gets Sarcsteroid B (3.3mg), the stream part when the collection retention time is 54min to doing, reclaim under reduced pressure gets Sarcsteroid D (5.4mg) to doing.
Fr.21 partly pass through Sephandex LH-20 gel column chromatography (moving phase normal hexane: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.21-1～Fr.21-7.Fr.21-5 part pass through again normal phase silica gel column chromatography (mobile phase methanol: wash-out chloroform=1: 50), detect monitoring according to thin layer plate, collect merge Fr.21～5-1～Fr.21-5-5.Fr.21-5-5 is by half preparation type RP-HPLC purifying (moving phase 95% methanol-water; Flow velocity 1.5ml/min; 30 ℃ of column temperatures), the differential detector monitors is collected the stream part when containing retention time and being 30min, and reclaim under reduced pressure is to doing, Sarcsteroid E (3.4mg), collect retention time stream part when being 38min, reclaim under reduced pressure is to doing, Sarcsteroid F (3.4mg).
Fr.22 partly pass through Sephandex LH-20 gel column chromatography (moving phase normal hexane: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.22-1～Fr.22-6.Fr.22-3 part pass through again Sephandex LH-20 gel column chromatography (moving phase normal hexane: methyl alcohol: wash-out chloroform=2: 1: 1), detect monitoring according to thin layer plate, collect merge Fr.22-3-1～Fr.22-3-6.Fr.22-3-5 by normal phase silica gel column chromatography (the moving phase chloroform: wash-out methyl alcohol=40: 1), again through more than half preparation type RP-HPLC purifying (moving phase 94% methanol-waters; Flow velocity 1.5ml/min; 30 ℃ of column temperatures), the differential detector monitors is collected the stream part when containing retention time and being 23min, and reclaim under reduced pressure is to doing, SarcsteroidC (1.4mg).
Embodiment 3. preparation Anthsteroids A～B
Get piece flower gorgonian (Anthogorgia sp) 2167g of surrounding waters, the Guangxi China North Sea, clean, shred, with the acetone supersound extraction of 5 times of weight, reclaim under reduced pressure acetone is used the equal-volume water-dispersion to there not being the acetone flavor, extracted with diethyl ether 6 times, each 2000ml, combining extraction liquid reclaims ether and is concentrated into dried.With the water suspendible of total medicinal extract with 2000ml, extract respectively 6 times with ether and propyl carbinol successively, merge and the reclaim under reduced pressure ether, get ether layer medicinal extract 28g, merge propyl carbinol and reclaim under reduced pressure propyl carbinol and get n-butanol layer medicinal extract 5g.Ether layer medicinal extract (28g) separates through normal phase silica gel column chromatography (200～300 order), with volume ratio is 99: 1,49: 1,34: 1,29: 1,24: 1,19: 1,14: 1,11: 1,9: 1,8: 1,7: 1,6: 1,5: 1,4: 1,3: 1,2: 1,1: 1, the sherwood oil/acetone gradient elution of 100% acetone, according to the thin layer plate monitoring, each flow point is collected as 32 part Fr.1～Fr.32 from small to large altogether by the polarity size.
Fr.21 partly passes through reversed-phase silica gel column chromatography (mobile phase methanol: water=50%, 70%, 80%, 90%) and does gradient elution, detects monitoring according to thin layer plate, collect merge Fr.21-1～Fr.21-6.Fr.21-5 is by half preparation type RP-HPLC purifying (moving phase 75% methanol-water; 30 ℃ of column temperatures; Flow velocity 1.5ml/min), differential detector monitors, the stream part when the collection retention time is 165min, reclaim under reduced pressure is to doing, get Anthsteroid A (1.2mg), the stream part when the collection retention time is 86min, reclaim under reduced pressure gets Anthsteroid B (2.1mg) to doing.
Embodiment 4: the in-vitro antibacterial test
One, fungi, bacterium and algae are used in test
1. fungi is used in test: anther smut (Microbotryum violaceum) and septoria musiva leaf spoting bacteria (Septoria tritici).
2. bacterium is used in test: intestinal bacteria (Escherichia coli), bacillus megaterium (Bacillus megaterium).
3. algae is used in test: chlorella (Chlorella fusca).
Two, experimental drug
1. positive control drug: penicillin (Penicillin), Streptomycin sulphate (Streptomycin), KETOKONAZOL.
2. negative control product: acetone (Acetone).
3.Muristeroids A～G, courage steroid-5 α, 6 beta-diols-3 β-acetic ester, courage steroid-5 α-methoxyl group-3 β, 6 beta-diols, (22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α, 6 beta-triols, courage steroid-1 β, 3 β, 5 α, 6 β-tetrol, Sarcsteroids A～F, Anthsteroids A～B is by embodiment 1 preparation
Three, experimental technique
1. compounding pharmaceutical solution: with penicillin, Streptomycin sulphate, KETOKONAZOL, Muristeroids A～G, courage steroid-5 α, 6 beta-diols-3 β-acetic ester, courage steroid-5 α-methoxyl group-3 β, 6 beta-diols, (22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α, 6 beta-triols, courage steroid-1 β, 3 β, 5 α, 6 β-tetrol, Sarcsteroids A～F, Anthsteroids A～B is mixed with the solution that concentration is 2mg/ml with acetone respectively, single test consumption 25 μ l.
2. above-mentioned 2 kinds of fungies, 2 kinds of bacteriums and a kind of algae are mixed with bacterium liquid with the 7ml aqua sterilisa respectively by the requirement of aseptic technique, each gets 4ml bacterium liquid with atomizer, evenly be sprayed on the media surface of culture dish separately respectively, in each culture dish, place two of the about 1cm sterilization of diameter filter paper more respectively, be covered in media surface, the soup 50 μ l that get above-mentioned preparation then respectively drip on filter paper, cover the substratum lid and cultivate.All indicate corresponding kinds of culture medium, bacterial classification, compound title, inoculation time on each culture dish lid.Anther smut was cultivated 4 days for 20 ℃, and the septoria musiva leaf spoting bacteria was cultivated 4 days for 20 ℃, and intestinal bacteria were cultivated 24 hours for 37 ℃, and bacillus megaterium was cultivated 24 hours for 37 ℃, and chlorella was cultivated 5 days for 20 ℃.Observations is on time measured the size (radius) of inhibition zone, and parallel test 3 times is averaged, and the results are shown in Table 16.
Table 16. agar diffusion experiment screening active ingredients (mm)
By table 16 as seen, Muristeroids A～G of the present invention, Sarcsteroids A～F, AnthsteroidsA～B, courage steroid-5 α, 6 beta-diols-3 β-acetic ester, courage steroid-5 α-methoxyl group-3 β, 6 beta-diols, (22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α, 6 beta-triols, courage steroid-1 β, 3 β, 5 α, 6 β-tetrol is to anther smut, the septoria musiva leaf spoting bacteria, Gram-negative bacteria-intestinal bacteria, gram-positive microorganism-bacillus megaterium and chlorella all have significant inhibitory effect, therefore can be used for preparing antibacterials.
The present invention provides new lead compound for developing new antibacterials, and is significant to the marine pharmaceutical organism resource that develops China.
Claims (2)
1. polyhydroxyl steroid compounds Muristeroids A～G, Sarcsteroids A～F, AnthsteroidsA～B is characterized in that their chemical structure of general formula is as follows:
The R of each compound wherein
1, R
2, R
3, R
4, R
5And R
6Group collocation as follows:
2. the application of polyhydroxyl steroid compounds in preparation antibacterials or anti-little algae medicine is characterized in that this compound is poly-hydroxy steroid compound Muristeroids A～G as claimed in claim 1, Sarcsteroids A～F, Anthsteroids A～B; And courage steroid-5 α, 6 beta-diols-3 β-acetic ester, courage steroid-5 α-methoxyl group-3 β, 6 beta-diols, (22E)-22-alkene-courage steroid-3 β, 5 α, 6 beta-triols, 24 (28)-alkene-ergot steroid-3 β, 5 α, 6 beta-triols, courage steroid-3 β, 5 α, 6 beta-triols, or courage steroid-1 β, 3 β, 5 α, 6 β-tetrol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110119322 CN102250181B (en) | 2011-05-10 | 2011-05-10 | Polyhydroxy steroid compounds and purpose thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110119322 CN102250181B (en) | 2011-05-10 | 2011-05-10 | Polyhydroxy steroid compounds and purpose thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102250181A true CN102250181A (en) | 2011-11-23 |
| CN102250181B CN102250181B (en) | 2013-06-12 |
Family
ID=44977783
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110119322 Expired - Fee Related CN102250181B (en) | 2011-05-10 | 2011-05-10 | Polyhydroxy steroid compounds and purpose thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102250181B (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103214536A (en) * | 2013-02-07 | 2013-07-24 | 中国人民解放军第二军医大学 | Polyhydroxy steroidal compounds separated from coral, and application thereof |
| CN104844438A (en) * | 2014-02-18 | 2015-08-19 | 中国人民解放军第二军医大学 | Compounds with tetracyclic fused ring structure, and preparation method and application thereof |
| CN105837650A (en) * | 2016-04-05 | 2016-08-10 | 中国人民解放军第二军医大学 | Steroid compound and application thereof |
| CN107822912A (en) * | 2017-10-23 | 2018-03-23 | 陈有平 | Compound titanii dioxydi gel composition |
| CN107822913A (en) * | 2017-10-23 | 2018-03-23 | 陈有平 | Compound external-use gel containing titanium dioxide |
| CN107823219A (en) * | 2017-10-23 | 2018-03-23 | 陈有平 | Compound external-use gel combination |
| CN107823218A (en) * | 2017-10-23 | 2018-03-23 | 陈有平 | Suppress the Compound titanii dioxydi gel composition of light aging |
| CN108135909A (en) * | 2015-05-18 | 2018-06-08 | 布莱阿姆青年大学 | Cationic steroidal Antimicrobe compound and the method for preparing such compound |
-
2011
- 2011-05-10 CN CN 201110119322 patent/CN102250181B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| 《Chemical & Pharmaceutical Bulletin》 20100629 Sung Ping-Jyun, et al. New 3beta,5alpha,6beta-Trihydroxysteroids from the Octocorals Bebryce sp. (Plexauridae) and Carijoa sp. (Clavulariidae) 第1240-1242页 1 第58卷, 第9期 * |
| 《中国博士学位论文全文数据库-医药卫生科技辑》 20091015 王增蕾 仿刺参和一种肉芝软珊瑚化学成分及生物活性研究 第E079-8页 1 , 第10期 * |
| 《药学实践杂志》 20100725 孟丽媛等 短指软珊瑚中的生态活性物质 第274-278页 1-2 第28卷, 第4期 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103214536A (en) * | 2013-02-07 | 2013-07-24 | 中国人民解放军第二军医大学 | Polyhydroxy steroidal compounds separated from coral, and application thereof |
| CN103214536B (en) * | 2013-02-07 | 2015-12-09 | 中国人民解放军第二军医大学 | The polyhydroxysteroids compounds and application thereof that obtain is separated from coral |
| CN104844438A (en) * | 2014-02-18 | 2015-08-19 | 中国人民解放军第二军医大学 | Compounds with tetracyclic fused ring structure, and preparation method and application thereof |
| WO2015124037A1 (en) * | 2014-02-18 | 2015-08-27 | 中国人民解放军第二军医大学 | Compound with four-ring fused structure, preparation method therefor and application thereof |
| CN104844438B (en) * | 2014-02-18 | 2017-05-17 | 中国人民解放军第二军医大学 | Compounds with tetracyclic fused ring structure, and preparation method and application thereof |
| CN108135909A (en) * | 2015-05-18 | 2018-06-08 | 布莱阿姆青年大学 | Cationic steroidal Antimicrobe compound and the method for preparing such compound |
| CN105837650A (en) * | 2016-04-05 | 2016-08-10 | 中国人民解放军第二军医大学 | Steroid compound and application thereof |
| CN107822912A (en) * | 2017-10-23 | 2018-03-23 | 陈有平 | Compound titanii dioxydi gel composition |
| CN107822913A (en) * | 2017-10-23 | 2018-03-23 | 陈有平 | Compound external-use gel containing titanium dioxide |
| CN107823219A (en) * | 2017-10-23 | 2018-03-23 | 陈有平 | Compound external-use gel combination |
| CN107823218A (en) * | 2017-10-23 | 2018-03-23 | 陈有平 | Suppress the Compound titanii dioxydi gel composition of light aging |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102250181B (en) | 2013-06-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102250181B (en) | Polyhydroxy steroid compounds and purpose thereof | |
| CN103665071B (en) | Elaiophylin derivative and the application in antimicrobial agent and resistance m tuberculosis infection thereof | |
| CN108640968B (en) | Mixed-source terpenoid and application thereof in preparation of anti-inflammatory drugs | |
| CN101362784A (en) | Ebomycin glycosides compounds, composition using the same as activity component and use thereof | |
| CN100494189C (en) | Benzofurantone compound, its production and use | |
| CN103214536B (en) | The polyhydroxysteroids compounds and application thereof that obtain is separated from coral | |
| CN110407792B (en) | Secalonic acid J compound derived from penicillium oxalicum and preparation method thereof | |
| CN102603818B (en) | Preparation method and use of cerebroside compounds | |
| CN107974412A (en) | A kind of penicillium roqueforti for being used to prepare antiinflammatory active compound peniroquesine A and its application | |
| CN109988129A (en) | Compound and its preparing the application in antituberculotic | |
| CN102229588B (en) | Briarane type diterpenoid compounds with anti-tumor and antibacterial activities and application thereof | |
| CN109970687A (en) | A kind of terpene derivant and the preparation method and application thereof | |
| CN103145740B (en) | Sulfoxide alkaloid compound as well as preparation method and application for same | |
| CN107522605A (en) | Sequiterpene methyl cyclopentene diketone, its preparation method and its application | |
| CN110407794B (en) | Selenolonic acid K derived from penicillium oxalicum and application thereof in inhibiting cancer cell proliferation | |
| CN101863936B (en) | Open loop steroid saponin compound with antibacterial activity in sinularia gyrosa | |
| CN110407797B (en) | Secalonic acid K compound derived from penicillium oxalicum and preparation method thereof | |
| CN107815473B (en) | Diphenyl ether compound and preparation method and application thereof | |
| CN101235040B (en) | Phomopsis rhzomorph compound and its preparation method and application | |
| CN109956883A (en) | A kind of acetylation flores aurantii containing azo-cycle alkane type sesquiterpene derivative and its preparation and application | |
| CN105218495A (en) | A kind of red sage root water soluble ingredient new compound, preparation method and application thereof | |
| CN114573537B (en) | Compound with anti-neurodegenerative change activity, preparation method and application thereof | |
| CN110407795B (en) | Penicillium oxalicum ketoacid L and application thereof in inhibiting human cancer cell proliferation | |
| CN110403929B (en) | Selenolonic acid M derived from penicillium oxalicum and application thereof in inhibiting human cancer cell proliferation | |
| CN110357864B (en) | Pyrrole sesquiterpenoid compound and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130612 |