CN104926916A - Steroid, medicine composition containing novel steroid I, as well as preparation method and application of medicine composition - Google Patents
Steroid, medicine composition containing novel steroid I, as well as preparation method and application of medicine composition Download PDFInfo
- Publication number
- CN104926916A CN104926916A CN201510301493.7A CN201510301493A CN104926916A CN 104926916 A CN104926916 A CN 104926916A CN 201510301493 A CN201510301493 A CN 201510301493A CN 104926916 A CN104926916 A CN 104926916A
- Authority
- CN
- China
- Prior art keywords
- preparation
- sah
- medicine composition
- steroid
- separated
- 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
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 239000003814 drug Substances 0.000 title abstract description 8
- 239000000203 mixture Substances 0.000 title abstract description 6
- 150000003431 steroids Chemical class 0.000 title abstract description 6
- 230000000840 anti-viral effect Effects 0.000 claims abstract description 9
- 150000001875 compounds Chemical class 0.000 claims description 31
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical class CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 11
- 239000008194 pharmaceutical composition Substances 0.000 claims description 10
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 239000000284 extract Substances 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 7
- 238000000605 extraction Methods 0.000 claims description 7
- 239000000741 silica gel Substances 0.000 claims description 7
- 229910002027 silica gel Inorganic materials 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 239000002027 dichloromethane extract Substances 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 5
- 230000003637 steroidlike Effects 0.000 claims description 5
- 239000003937 drug carrier Substances 0.000 claims description 4
- 238000000746 purification Methods 0.000 claims description 4
- 230000002441 reversible effect Effects 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 3
- FDSGHYHRLSWSLQ-UHFFFAOYSA-N dichloromethane;propan-2-one Chemical compound ClCCl.CC(C)=O FDSGHYHRLSWSLQ-UHFFFAOYSA-N 0.000 claims description 3
- 239000012259 ether extract Substances 0.000 claims description 3
- 239000000499 gel Substances 0.000 claims description 3
- 238000010829 isocratic elution Methods 0.000 claims 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 claims description 3
- 239000003208 petroleum Substances 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 13
- 108020002202 Adenosylhomocysteinase Proteins 0.000 abstract description 4
- 102000005234 Adenosylhomocysteinase Human genes 0.000 abstract description 3
- 238000000338 in vitro Methods 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 abstract description 2
- 238000000926 separation method Methods 0.000 abstract 1
- ZJUKTBDSGOFHSH-WFMPWKQPSA-N S-Adenosylhomocysteine Chemical compound O[C@@H]1[C@H](O)[C@@H](CSCC[C@H](N)C(O)=O)O[C@H]1N1C2=NC=NC(N)=C2N=C1 ZJUKTBDSGOFHSH-WFMPWKQPSA-N 0.000 description 31
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 20
- 101000925662 Enterobacteria phage PRD1 Endolysin Proteins 0.000 description 17
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 15
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 15
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 12
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 12
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 10
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 10
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 9
- 229960005305 adenosine Drugs 0.000 description 9
- FFFHZYDWPBMWHY-VKHMYHEASA-N L-homocysteine Chemical compound OC(=O)[C@@H](N)CCS FFFHZYDWPBMWHY-VKHMYHEASA-N 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 8
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 7
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 7
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 6
- 241000208688 Eucommia Species 0.000 description 6
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 6
- 235000019253 formic acid Nutrition 0.000 description 6
- 230000003301 hydrolyzing effect Effects 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 229910052500 inorganic mineral Inorganic materials 0.000 description 6
- FEWJPZIEWOKRBE-JCYAYHJZSA-L L-tartrate(2-) Chemical compound [O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O FEWJPZIEWOKRBE-JCYAYHJZSA-L 0.000 description 5
- 241000700605 Viruses Species 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 235000015165 citric acid Nutrition 0.000 description 5
- 108020004999 messenger RNA Proteins 0.000 description 5
- 239000011707 mineral Substances 0.000 description 5
- 150000007524 organic acids Chemical class 0.000 description 5
- 235000006408 oxalic acid Nutrition 0.000 description 5
- 150000003839 salts Chemical class 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 229940095064 tartrate Drugs 0.000 description 5
- FFFHZYDWPBMWHY-UHFFFAOYSA-N L-Homocysteine Natural products OC(=O)C(N)CCS FFFHZYDWPBMWHY-UHFFFAOYSA-N 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 239000003981 vehicle Substances 0.000 description 4
- 239000002775 capsule Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000008187 granular material Substances 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 239000008057 potassium phosphate buffer Substances 0.000 description 3
- 239000011541 reaction mixture Substances 0.000 description 3
- 230000000452 restraining effect Effects 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 208000032843 Hemorrhage Diseases 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 206010037660 Pyrexia Diseases 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- -1 lignanoids Chemical class 0.000 description 2
- 150000002894 organic compounds Chemical class 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 206010000242 Abortion threatened Diseases 0.000 description 1
- 102100020925 Adenosylhomocysteinase Human genes 0.000 description 1
- 208000008035 Back Pain Diseases 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000208686 Eucommiaceae Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 206010020112 Hirsutism Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 229940124036 Hydrolase inhibitor Drugs 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- 208000008930 Low Back Pain Diseases 0.000 description 1
- 241000218922 Magnoliophyta Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000016397 Methyltransferase Human genes 0.000 description 1
- 108060004795 Methyltransferase Proteins 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000009328 Perro Species 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- MEFKEPWMEQBLKI-AIRLBKTGSA-N S-adenosyl-L-methioninate Chemical compound O[C@@H]1[C@H](O)[C@@H](C[S+](CC[C@H](N)C([O-])=O)C)O[C@H]1N1C2=NC=NC(N)=C2N=C1 MEFKEPWMEQBLKI-AIRLBKTGSA-N 0.000 description 1
- 208000005985 Threatened Abortion Diseases 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 108010067390 Viral Proteins Proteins 0.000 description 1
- XEDIUOFTIAIXCV-UHFFFAOYSA-N [N+](=O)([O-])C1=C(C(=O)O)C=CC=C1.[S] Chemical compound [N+](=O)([O-])C1=C(C(=O)O)C=CC=C1.[S] XEDIUOFTIAIXCV-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 150000003835 adenosine derivatives Chemical class 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 238000012870 ammonium sulfate precipitation Methods 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 150000001721 carbon Chemical group 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000001952 enzyme assay Methods 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 239000004093 hydrolase inhibitor Substances 0.000 description 1
- 208000021822 hypotensive Diseases 0.000 description 1
- 230000001077 hypotensive effect Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 230000002101 lytic effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000002366 mineral element Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000005311 nuclear magnetism Effects 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 238000005453 pelletization Methods 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- CRUSHRATSSNWSB-UHFFFAOYSA-N phenyl-sulfanylidene-sulfidoazanium Chemical compound [S-][N+](=S)C1=CC=CC=C1 CRUSHRATSSNWSB-UHFFFAOYSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000008227 sterile water for injection Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J71/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton is condensed with a heterocyclic ring
- C07J71/0005—Oxygen-containing hetero ring
- C07J71/001—Oxiranes
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the technical field of medicine and particularly relates to a novel steroid I obtained from folium cortex eucommiae through separation, a medicine composition containing the novel steroid I, as well as a preparation method and application of the medicine composition. The novel steroid I is reported for the first time, can restrain activity of SAH hydrolase as proved by in vitro test, may have the antiviral activity, and can be applied in the antiviral field.
Description
Technical field
The invention belongs to medical art, be specifically related to from Folium Eucommiae, be separated a kind of new steroidal compounds obtained, containing its pharmaceutical composition and its preparation method and application.
Background technology
The bark of eucommia
eucommia ulmoidesoliv. belong to Angiospermae, Eucommiaceae, the bark of eucommia belongs to, deciduous tree, up to 20m, and bark beige, together with branch, leaf, root all containing glue, the silvery white that fractureed filament.Elliptic leaf or oval volt ovum type, long 6 ~ 18cm, there is sawtooth at edge, hairiness on following arteries and veins, and the long 1 ~ 2cm of petiole, really for samara is flat and thin, includes a seed.The bark of eucommia has the medicinal history of more than 2000 year in China as Chinese medicine, Shennong's Herbal and Compendium of Material Medica are all classified as top grade, and it has multiple efficacies, is mainly used in treating lumbago due to renal deficiency, and muscles and bones is unable, blood leaking in gestation, threatened abortion, vascular hypertension.
In recent years, scholars has carried out large quantity research to chemical component of eucommia bark used, is separated at present to obtain and organic compound through qualification has more than 70 kinds, and inorganic mineral element is no less than 15 kinds.Research finds, containing similar chemical composition in each several parts such as Cortex Eucommiae, flower, leaf and branch, mainly comprises the organic compound such as lignanoids, iridoids, flavonoid and steroid.Modern pharmacology proves, the bark of eucommia has several functions activity, as hypotensive, anti-ageing, anticancer, antibacterial, analgesia, anti-inflammatory, diuresis, calmness, to calm the nerves, antivirus action, enhancing body nonspecific immunity power, and the effects such as dual modulation can be carried out to cellular immunization, also there is certain effect to nervous center in addition.The bark of eucommia is also because its significant physiologically active obtains the concern of numerous scholar.
Modern study shows, some plant not only has the immunologic function regulating body, also has nonspecific antiviral functions.S-adenosyl-L-homocysteine lytic enzyme (S-Adenosyl-L-homocysteine hydrolase, SAH lytic enzyme) be the effect target that a class has the adenosine derivative of unique broad-spectrum antiviral activity, especially the object target of hemorrhage fever virus inhibitor, extensively be present in vertebrates (as people, ox, dog, rabbit, rat, mouse), plant, fungi and other microorganism cellss, there is important physiological function.SAH lytic enzyme and transmethylation closely related, eukaryotic cell and virus mRNA 5 '-end must have methylated cap-like structure, just can translate.MRNA 5 '-terminal methyl group depends on the multiple methyl transferase catalytic of adenosylmethionine, and SAH is enzyme reaction product, is also the competitive feedback inhibition agent of this enzyme.SAH can be hydrolyzed into adenosine and L-homocysteine by SAH lytic enzyme, and this reaction is reversible, external trend synthesis, trend hydrolysis in body.The SAH of transmethylated generation in body is constantly hydrolyzed by SAH lytic enzyme, transmethylase is reacted and proceeds, without the accumulation of SAH.As suppressed in SAH lytic enzyme, SAH accumulation in cell, suppress methylferase, and then affect mRNA function, viral protein can not synthesize, and also just can not generate progeny virus, virus replication is suppressed.Most of virus mRNA needs methylated cap-like structure, and responsive to the rejection ratio cell mRNA of methylferase, and in cell, SAH concentration has increased slightly, and is just first involved, and it is selective that this feature gives SAH hydrolase inhibitor.Therefore, SAH lytic enzyme is used as the in-vitro screening model of screening hemorrhage fever virus inhibitor in the world.
Summary of the invention
The object of this invention is to provide and a kind ofly from Folium Eucommiae, be separated a kind of new steroidal compounds (I) obtained, its pharmaceutical composition and its production and use.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
There is the steroidal compounds (I) of following structural formula,
Pharmaceutical composition, the compound (I) wherein containing treatment significant quantity and pharmaceutically acceptable carrier.
The preparation method of described compound (I), is characterized in that, comprise following methods step: the Folium Eucommiae that (a) dries in the shade 90% alcohol heat reflux extracts 3 times, and united extraction liquid is concentrated into without alcohol taste, obtains concentrated solution; B the concentrated solution of step (a) gained is used sherwood oil, methylene dichloride and water saturated n-butanol extraction by () successively, obtain petroleum ether extract, dichloromethane extract and n-butyl alcohol extract respectively; C () is separated dichloromethane extract purification on normal-phase silica gel in step (b), obtain 5 components successively with the dichloromethane-acetone gradient elution that volume ratio is 50:1,20:1,10:1,5:1 and 1:1; D component 2 in step (c) reverse phase silica gel is separated by (), be the aqueous methanol gradient wash-out of 30%, 50%, 70% and 100% successively, obtain 4 components by concentration expressed in percentage by volume; E component 3 in step (d) dextrane gel Sephadex LH-20 is separated by (), be the methanol aqueous solution isocratic elution of 75%, obtain pure compound (I) by concentration expressed in percentage by volume.
The application of described compound (I) in preparation antiviral.
The application of described pharmaceutical composition in preparation antiviral.
When the compounds of this invention is used as medicine, directly can uses, or use with the form of pharmaceutical composition.This pharmaceutical composition contains the compounds of this invention (I) for the treatment of significant quantity, and all the other are acceptable on pharmacology, nontoxic to humans and animals and pharmaceutically acceptable carrier of inertia and/or vehicle.
Described pharmaceutically acceptable carrier or vehicle are that one or more are selected from solid, semisolid and liquid diluent, filler and pharmaceutical preparation assistant agent.Pharmaceutical composition of the present invention is used with the form of per weight dose.Medicine of the present invention is applied to by form that is oral or injection the patient needing treatment.For time oral, tablet, slow releasing tablet, controlled release tablet, capsule, dripping pill, micropill, suspensoid, emulsion, powder or granule, oral liquid etc. can be made into; During for injecting, can be made into water-based or oily solution, aseptic powder injection, liposome or the emulsion etc. of sterilizing.
Accompanying drawing explanation
Fig. 1 is compound (I) chemical structural formula;
Fig. 2 is blank curve and SAH hydrolytic enzyme activities measurement result;
Fig. 3 is that compound (I) is to the restraining effect of SAH lytic enzyme.
Embodiment
Further illustrate essentiality content of the present invention below in conjunction with embodiment, but do not limit scope with this.Although be explained in detail the present invention with reference to preferred embodiment, those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the scope of technical solution of the present invention.
Main raw and reagent: ethanol, sherwood oil, methylene dichloride, propyl carbinol, acetone are analytical pure, purchased from Nanjing Chemistry Reagent Co., Ltd., methyl alcohol, analytical pure, purchased from Hanbon Sci. & Tech. Co., Ltd.; Ado, Sigma company; L-Hey, Sigma company; DTNB, Sigma company; Dithiothreitol (DTT) (DTT), analytical pure, Bai Ao bio tech ltd, Shanghai; Ethylenediamine tetraacetic acid (EDTA) (EDTA), analytical pure, Tianjin oceangoing voyage Chemical Company; Ammonium sulfate, analytical pure, the huge chemical reagent factory in Dongli District, Tianjin; Potassium primary phosphate, analytical pure, the huge chemical reagent factory in Dongli District, Tianjin; Dipotassium hydrogen phosphate, analytical pure, the huge chemical reagent factory in Dongli District, Tianjin; Wistar rat, Harbin Medical University's Experimental Animal Center.Folium Eucommiae originates from Jiangsu great Feng Forestry Base.
Nuclear magnetic resonance analyser (German Bruker company); WRS-1B type digital melting point determinator (Shanghai precision instrument science company limited); WatersSynaptTMQ-TOF type mass spectrograph (Waters, US); The automatic purification system of Waters2545-2767-UV2489 (Waters, US); Thin-layer chromatography and column chromatography silica gel (Haiyang Chemical Plant, Qingdao).
Embodiment 1: the preparation method of compound (I)
Extraction and isolation step: the Folium Eucommiae (10.5 Kg) that (a) dries in the shade extracts with 90% alcohol heat reflux, extracts three times, each 30L, and extract 2.5 hours, united extraction liquid, is concentrated into without alcohol taste, obtains concentrated solution; B () step (a) gained concentrated solution uses sherwood oil (2L), methylene dichloride (2L) and water saturated propyl carbinol (2L) to extract successively, obtain petroleum ether extract, dichloromethane extract (265g) and n-butyl alcohol extract; C () is separated the dichloromethane extract purification on normal-phase silica gel in step (b), be the dichloromethane-acetone gradient elution of 50:1,20:1,10:1,5:1 and 1:1 successively by volume ratio, each gradient elution 6 column volumes, merge that identical gradient eluent is concentrated obtains 5 components; D () is by the component 2(78g in step (c)) be separated with reverse phase silica gel, be the aqueous methanol gradient wash-out of 30%, 50%, 70% and 100% successively by concentration expressed in percentage by volume, each gradient elution 5 column volumes, merge that identical gradient eluent is concentrated obtains 4 components; E () is by the component 3(5.4g in step (d)) be separated with dextrane gel Sephadex LH-20, be the methanol aqueous solution isocratic elution of 75% by concentration expressed in percentage by volume, obtain pure compound (I) (27mg).
Structural identification: the crystallization of white needles; HR-ESIMS shows [M+Na]
+for m/z 461.3424, can obtain molecular formula in conjunction with nuclear-magnetism feature is C
30h
46o
2, degree of unsaturation is 8.Hydrogen nuclear magnetic resonance modal data δ H(ppm, CDCl
3, 600MHz): 1.32(1H, m, 1-H), 1.08(1H, m, 1-H), 1.53(1H, m, 2-H), 1.28(1H, m, 2-H), 3.54(1H, m, 3-H), 2.22(1H, dd, 4-H), 1.97(1H, dd, 4-H), 5.28(1H, dd, 6-H), 2.19(1H, m, 7-H), 1.95(1H, m, 7-H), 1.49(1H, m, 8-H), 1.69(1H, dd, 9-H), 5.91(1H, dd, 11-H), 5.59(1H, d, 12-H), 1.45(1H, dd, 14-H), 2.25(1H, dd, 15-H), 2.25(1H, dd, 16-H), 1.45(1H, dd, 17-H), 1.05(3H, s, 18-H), 1.05(3H, s, 19-H), 1.21(1H, m, 20-H), 0.89(3H, d, 21-H), 1.13(1H, m, 22-H), 1.11(1H, m, 24-H), 1.42(1H, m, 25-H), 0.84(3H, d, 26-H), 0.84(3H, d, 27-H), 0.89(3H, d, 28-H), 0.85(3H, s, 29-H), 0.81(1H, dd, 30-H), 0.26(1H, dd, 30-H), 4.78(1H, s, 3-OH), carbon-13 nmr spectra data δ C(ppm, CDCl
3, 150MHz): 38.0(CH
2, 1-C), 31.9(CH
2, 2-C), 71.7(CH, 3-C), 41.9(CH
2, 4-C), 142.4(C, 5-C), 121.9(CH, 6-C) and, 25.9(CH
2, 7-C), 27.4(CH, 8-C), 56.6(CH, 9-C) and, 41.6(C, 10-C), 131.4(CH, 11-C), 138.7(CH, 12-C), 29.5(C, 13-C) and, 59.8(CH, 14-C), 61.4(CH, 15-C), 61.1(CH, 16-C), 59.4(CH, 17-C) and, 20.9(CH
3, 18-C), 19.1(CH
3, 19-C), carbon atom mark is see accompanying drawing 1.
Embodiment 2: the activity test of compound (I)
1, experimental technique
The extraction of 1.1 SAH lytic enzymes
Get fresh rat liver at 4 DEG C, make homogenate with 2 times of 10mmol/L pH7.6 phosphoric acid buffers to liver weight (containing 2mmol/L DTT, 1mmol/L EDTA).Liver pulp is at 4 DEG C 10, and the centrifugal 30min of 000 rad/min, gets supernatant liquor, 4 DEG C 10, and the centrifugal 60min of 000 rad/min, gets supernatant liquor, and supernatant liquor 38% ammonium sulfate precipitation, in 4 DEG C 10, after 000 rad/min is centrifugal, abandons supernatant, get precipitation freeze-drying for subsequent use.
1.2 SAH hydrolytic enzyme activities measure
By the activity of colorimetric method for determining SAH lytic enzyme.SAH lytic enzyme reversibly can be decomposed into adenosine (Ado), homocysteine (Hcy) by catalysis S-adenosyl-L-homocysteine (SAH), can there is thio-disulfide permutoid reaction with dithio-nitrobenzene formic acid (DTNB) in halfcystine, in reaction, the halfcystine of 1 molecule causes the release of the sulphur nitrobenzoic acid of 1 molecule.It is when PH 8.0, has strong absorption at 412 nm wavelength places, and colorimetric determination therefore can be utilized to measure-SH base, measures absorb light angle value to detect SAH hydrolytic enzyme activities.
Blank curve plotting: by 100 μ l reaction mixtures (containing DTT 2mM, EDTA 1mM, adenosine 2mM, D, L-homocysteine 2mM, potassium phosphate buffer pH 7.6 120mM) 6 parts, cultivate 0 second in 37 DEG C of water-baths respectively, 10 seconds, 20 seconds, 30 seconds, 40 seconds, 50 seconds, then 20 μ l are respectively got, join respectively in the centrifuge tube filling 60 μ l Tris-Cl damping fluids (PH 8.0) and mix, respectively get 20 μ l again to join 6 respectively and fill in the centrifuge tube of 100 μ l DTNB analytic liquids, accurately leave standstill 5 min, it is made fully to develop the color, light absorption value is detected at 412nm place.
SAH hydrolytic activity curve plotting: by 100 μ l reaction mixtures (containing DTT 2mM, EDTA 1mM, adenosine 2mM, D, L-homocysteine 2mM, potassium phosphate buffer pH 7.6 120mM, SAH lytic enzyme 3 μ l) 6 parts, experimental technique is the same.
1.3 compounds (I) are to the restraining effect of SAH hydrolytic enzyme activities
Compound (I) distilled water is mixed with the sample solution that concentration is 0.05mg/ml.By 100 μ l reaction mixtures (containing DTT 2mM, EDTA 1mM, adenosine 2mM, D, L-homocysteine 2mM, potassium phosphate buffer pH 7.6 120mM, SAH lytic enzyme 3 μ l, compound (I) sample solution 3 μ l) 6 parts, experimental technique is the same.
2 results and analysis
2.1 blank curves and SAH hydrolytic enzyme activities measure
Visible in Fig. 2, reaction substrate adenosine and homocysteine, can slowly to the orienting responses of synthesis when not adding SAH lytic enzyme, and in blank curve, the absorbancy of substrate reduces gradually.SAH lytic enzyme is the catalyzer of this reaction, when substrate adenosine and homocysteine concentration very high, SAH Hydrolases catalyze adenosine and homocysteine react rapidly, but after 10 seconds, this Reactive Synthesis and hydrolysis reach balance, and in reaction system, the amount of homocysteine tends towards stability.In Fig. 2, Enzyme assay curve first 10 seconds substrate absorbancys reduce rapidly, and after 10 seconds, the absorbancy of substrate tends towards stability gradually, and reaction just reaches balance.
2.2 compounds (I) are to the restraining effect of SAH lytic enzyme
Visible in Fig. 3, after adding compound (I), adenosine and homocysteine continue the orienting response to synthesis, do not occur that the accelerated reaction of SAH lytic enzyme also reaches rapidly the phenomenon of molecular balance.Illustrate that compound (I) inhibits the activity of SAH lytic enzyme.Analysis may have antiviral activity.
Embodiment 3:
The preparation of tablet: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, vehicle is added, pelletizing press sheet than the ratio for 1:7 in itself and excipient weight.
Embodiment 4:
The preparation of oral liquid: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, oral liquid method for making makes oral liquid routinely.
Embodiment 5:
The preparation of capsule or granule: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, add vehicle in itself and excipient weight than the ratio for 1:8.5, make capsule or granule.
Embodiment 6:
The preparation of injection liquid: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, inject with water routinely, essence filter, injection liquid is made in embedding sterilizing.
Embodiment 7:
The preparation of aseptic powder injection: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, be dissolved in sterile water for injection, stirring makes molten, filter with aseptic suction funnel, aseptic essence filter again, be sub-packed in ampoule, after frozen drying, aseptic sealing by fusing obtains powder injection.
Claims (4)
1. there is the steroidal compounds (I) of following structural formula,
Pharmaceutical composition, the compound according to claim 1 (I) wherein containing treatment significant quantity and pharmaceutically acceptable carrier.
2. the preparation method of compound according to claim 1 (I), is characterized in that, comprises following methods step: the Folium Eucommiae that (a) dries in the shade 90% alcohol heat reflux extracts 3 times, and united extraction liquid is concentrated into without alcohol taste, obtains concentrated solution; B the concentrated solution of step (a) gained is used sherwood oil, methylene dichloride and water saturated n-butanol extraction by () successively, obtain petroleum ether extract, dichloromethane extract and n-butyl alcohol extract respectively; C () is separated dichloromethane extract purification on normal-phase silica gel in step (b), obtain 5 components successively with the dichloromethane-acetone gradient elution that volume ratio is 50:1,20:1,10:1,5:1 and 1:1; D component 2 in step (c) reverse phase silica gel is separated by (), be the aqueous methanol gradient wash-out of 30%, 50%, 70% and 100% successively, obtain 4 components by concentration expressed in percentage by volume; E component 3 in step (d) dextrane gel Sephadex LH-20 is separated by (), be the methanol aqueous solution isocratic elution of 75%, obtain pure compound (I) by concentration expressed in percentage by volume.
3. the application of compound according to claim 1 (I) in preparation antiviral.
4. the application of pharmaceutical composition according to claim 2 in preparation antiviral.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510301493.7A CN104926916B (en) | 2015-06-05 | 2015-06-05 | Steroidal compounds, containing its pharmaceutical composition and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510301493.7A CN104926916B (en) | 2015-06-05 | 2015-06-05 | Steroidal compounds, containing its pharmaceutical composition and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104926916A true CN104926916A (en) | 2015-09-23 |
| CN104926916B CN104926916B (en) | 2016-08-24 |
Family
ID=54114360
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510301493.7A Expired - Fee Related CN104926916B (en) | 2015-06-05 | 2015-06-05 | Steroidal compounds, containing its pharmaceutical composition and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104926916B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1993003732A1 (en) * | 1991-08-13 | 1993-03-04 | Cocensys, Inc. | Gaba receptor modulators |
| WO2000063228A1 (en) * | 1999-04-15 | 2000-10-26 | Schering Aktiengesellschaft | Ent-steroids as selectively active estrogens |
-
2015
- 2015-06-05 CN CN201510301493.7A patent/CN104926916B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1993003732A1 (en) * | 1991-08-13 | 1993-03-04 | Cocensys, Inc. | Gaba receptor modulators |
| WO2000063228A1 (en) * | 1999-04-15 | 2000-10-26 | Schering Aktiengesellschaft | Ent-steroids as selectively active estrogens |
Non-Patent Citations (1)
| Title |
|---|
| 闫朝辉: "卫矛化学成分研究", 《中国优秀硕士学位论文全文数据库》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104926916B (en) | 2016-08-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Maas et al. | An unusual dimeric guaianolide with antiprotozoal activity and further sesquiterpene lactones from Eupatorium perfoliatum | |
| CN103508919B (en) | Alkaloid compound and preparing the purposes in anticomplement medicament | |
| Qi et al. | Chemical constituents from leaves of Camellia nitidissima and their potential cytotoxicity on SGC7901 cells | |
| CN105061451A (en) | Highly oxidized diterpenoid compound and preparation method and medical application thereof | |
| Siddiqui et al. | Optimization of ultrasound-assisted parthenolide extraction from Tarchonanthus camphoratus leaves using response surface methodology: HPTLC and cytotoxicity analysis | |
| CN105218617A (en) | A kind of new triterpenoid and preparation method thereof and medicinal use | |
| CN107936069B (en) | Coagulation-promoting apple flower effective component and extraction and separation method and application thereof | |
| CN101745024B (en) | Use of Nucleoside isoborneol in controlling quality for injection of red sage and dwarf lilyturf root | |
| CN105566427A (en) | Lanostane triterpene compound, and preparation method and medicinal use thereof | |
| CN104926916A (en) | Steroid, medicine composition containing novel steroid I, as well as preparation method and application of medicine composition | |
| CN105503995A (en) | Novel triterpenoid compound as well as preparation method and medical application thereof | |
| CN106176782A (en) | Herba Ecliptae chemical composition is as the purposes of phytoestrogen | |
| Zaghloul et al. | Taxodione, a DNA-binding compound from Taxodium distichum L.(Rich.) | |
| CN105461732A (en) | Novel diterpenoid compound and preparation method and medical application thereof | |
| CN101747190A (en) | Bisabolane sesquiterpene compound and preparation method and application thereof | |
| Du et al. | C-methyl flavonoid from the leaves of Cleistocalyx conspersipunctatus: α-glucosidase inhibitory, molecular docking simulation and biosynthetic pathway | |
| CN105418719A (en) | Lanostane triterpenoid for treating acute T-lymphocyte leukemia | |
| Ata | Novel plant-derived biomedical agents and their biosynthetic origin | |
| CN106749476A (en) | A kind of new liquor-saturated eggplant lactone compound and preparation method thereof and medical usage | |
| CN106606506A (en) | Use of enantio-labdane-type diterpene compounds in preparation of anti-complement drugs | |
| Khishova et al. | Quantitative determination of procyanidins in hawthorn fruits | |
| CN105520926A (en) | Uses of lignan compound (7S,8R)-dihydrodehydrodiconiferyl alcohol in preparation of anti-complement drugs | |
| CN105985347B (en) | A kind of benzo Macrocyclic lactams indole alkaloid and its purposes in anticomplement medicament is prepared | |
| Wang et al. | Four new tirucallane-type triterpenoids from Sapindus mukorossi Gaertn. flowers induced neurite outgrowth in PC12 cells related to insulin-like growth factor 1 receptor/phosphoinositide 3-kinase/extracellular regulated protein kinase signaling pathway | |
| CN106349328A (en) | Novel lactone compound and preparation method and medical 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 | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20170612 Address after: Ten in Hebei city of Handan Province, Guangping County 057650 Pu Xiang Bei Si Lang Gu Cun No. 174 Patentee after: Qin Meiying Address before: Quangang District of Quanzhou city Fujian province 362801 lotus pond community No. 723 CuO wa Patentee before: Zheng Zulan |
|
| CP02 | Change in the address of a patent holder | ||
| CP02 | Change in the address of a patent holder |
Address after: 266700 Liu Jia Zhuang village, Mingcun Town, Pingdu City, Qingdao, Shandong Patentee after: Qin Meiying Address before: Ten in Hebei city of Handan Province, Guangping County 057650 Pu Xiang Bei Si Lang Gu Cun No. 174 Patentee before: Qin Meiying |
|
| 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: 20160824 Termination date: 20180605 |