CN102532156A - Swerilactones H-K, 1-4 and medicinal composition and application thereof - Google Patents

Swerilactones H-K, 1-4 and medicinal composition and application thereof Download PDF

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CN102532156A
CN102532156A CN2010105879695A CN201010587969A CN102532156A CN 102532156 A CN102532156 A CN 102532156A CN 2010105879695 A CN2010105879695 A CN 2010105879695A CN 201010587969 A CN201010587969 A CN 201010587969A CN 102532156 A CN102532156 A CN 102532156A
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lactone
herba swertiae
swertiae bimaculatae
hepatitis
pharmaceutical composition
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CN102532156B (en
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陈纪军
耿长安
马云保
张雪梅
罗杰
黄晓燕
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Haobang Pharmaceutical Co., Ltd., Yunnan
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Kunming Institute of Botany of CAS
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Abstract

The invention discloses a compound shown as a structural formula (I), i.e., swerilactones H-K, 1-4, a medicinal composition consisting of an effective amount of the compound 1-4 and a medicinal carrier or an excipient, preparation methods of a medicinal compound, i.e., swerilactones H-K (1-4) and the medicinal composition thereof, an application of the swerilactones H-K, 1-4 to treatment of human diseases, and particularly an application of the swerilactones H-K, 1-4 to preparation of a medicament for resisting virus B hepatitis and a medicament for treating virus B hepatitis.

Description

Herba Swertiae bimaculatae lactone H-K, its medical composition and its use
Technical field:
The invention belongs to technical field of pharmaceuticals, specifically, relate to Herba Swertiae bimaculatae lactone H-K; Contain its pharmaceutical composition for the treatment hepatitis B of effective constituent; Its preparation method, and the application in pharmacy, the particularly application in the anti-hepatitis B medicine of preparation.
Background technology:
Hepatitis B is hepatitis B virus (hepatitis B virus; HBV) cause, be to lead and cause a kind of communicable disease of multiple organ injury with the liver inflammatory lesion, wherein chronic viral hepatitis is the main diseases therefore that causes liver cirrhosis, liver cancer.According to The World Health Organization (WHO) statistics, the whole world has 2,000,000,000 people of surpassing to infect or is infecting HBV, and wherein about 400,000,000 people are HBV chronic infection person, the chronic hepatitis, liver cirrhosis and the primary hepatitis that have every year more than 100 ten thousand populations to die from hepatitis B to cause.China belongs to hepatitis B infected high popular district, and chronic viral hepatitis B cause of disease carrier surpasses 1.2 hundred million, about 2800-3000 ten thousand examples of chronic viral hepatitis B patient.Therefore the hepatitis B infection is serious world's public health problem, and the expense of treatment chronic hepatitis is very expensive, and every year, due to illness virus hepatitis caused direct economic loss to reach 30,000,000,000～50,000,000,000 Renminbi.
The course of infection of hepatitis B virus related generally to for 5 steps: combine, invade, transcribe, translation, gene replication and packing.At first virion is attached to liver cell through acceptor, sloughs adventitia behind the intrusion endochylema, forms core particle.Core particle moves to liver cell nuclear, HBV DNA core particle shelling and going out, in nucleus, form cccDNA (covalently closed circularDNA, cccDNA).CccDNA is the template of hepatitis B replication, is that liver cell continues infected key substance for a long time.CccDNA has 2 kinds of functions, the various members of the synthetic virion of the first, and like HBsAg (surface antigen constitutes adventitia), HBcAg (hepatitis virus c antigens, cAg constitute inner membrance), HBeAg (being secreted in the blood); It two is synthetic HBV DNA of future generation.Most of new synthetic HBV DNA is used for the packing of reovirion, and part then gets into liver cell nuclear again, becomes another source of cccDNA.The new synthetic HBV DNA of part at first is assembled into core particle with HBcAg, and then is assembled into complete HBV particle with HBsAg, discharges to the extracellular.Superfluous HBsAg is released into blood sometimes alone, forms microspheric form particle or cast particle.
Present multiple therapeutic modality comprises Hepatitis B virus vaccine, immunomodulator and anti-HBV medicine nucleoside analog etc., has been applied to treat HBV and has infected, and obtained certain curative effect.Hepatitis B virus vaccine can effectively prevent HBV to infect, yet invalid to the patient who infects HBV.Immunomodulator like alpha-interferon (IFN) etc., can show anti-HBV effect through improving body immunity, yet itself does not have antiviral activity.And Interferon, rabbit has tangible restraining factors at the treatment chronic hepatitis B: at first, it never obtains real success, only if the substantive immunoreation to HBV has appearred in patient before treatment.And spinoff is obvious, and before interferon-induced HBeAg serum conversion, hepatitis active unexpected one property crossed increases the weight of; Showing as ALT (alanine aminotransferase, ALT, alanine aminotransferase) raises; Can cause patient with liver cirrhosis generation liver failure, occur dead once in a while.Nucleoside medicine is a present internationally recognized curative effect anti-hepatic-B virus medicine preferably, all is nucleoside analog like lamivudine (lamivudine), adefovir ester (adefovir dipivoxiil), Entecavir (enticavir), Telbivudine (tibivudine), the tynofovir (tenofovir) that has gone on the market at present.During these medicines are tested, HBV virus there is remarkable restraining effect in external or body.Its mechanism of action is to become the triphosphoric acid compound in the medicine entering cell, through the competition of substrate, to the polysaccharase or the ThermoScript II inhibition of virus, finally suppresses synthetic, the viral propagation of viral DNA.The anti-HBV of nucleoside analog has characteristics such as evident in efficacy, easy administration, bioavailability is high, patient tolerability is good.Yet also have tangible weak point, single like action target spot, take for a long time and cause virus strain variation, produce resistance, can not thoroughly eradicate HBV, be prone to recurrence after the drug withdrawal.Present anti-HBV medicine far can not satisfy the needs that treatment HBV infects, so the Antihepatitis medicament that the development structure novelty has new role mechanism has very large market outlook.
China's traditional Chinese medicine aboundresources, medicinal history is long, and takes for a long time through human body and to show having determined curative effect, low toxicity or advantage such as nontoxic.Mile Swertia Herb is the peculiar anti-hepatitis Chinese medicine in Yunnan, is used to treat acute icterohepatitis for a long time among the people, have better curative effect, yet its anti-HBV active substance is still unclear.
So far; The report of no compound Herba Swertiae bimaculatae lactone H-K (shown in the formula I) in the prior art; Do not have its report yet, do not have Herba Swertiae bimaculatae lactone H-K and pharmaceutical composition thereof the application report in preparation or treatment hepatitis B medicine yet as the pharmaceutical composition of effective constituent.
Summary of the invention:
The object of the present invention is to provide one type of Herba Swertiae bimaculatae lactone H-K (1-4) shown in the new formula with pharmaceutical use (I); Contain the Herba Swertiae bimaculatae lactone H-K (I) of treatment hepatitis B significant quantity and the treatment hepatitis B pharmaceutical composition of pharmaceutical carrier or vehicle; Herba Swertiae bimaculatae lactone H-K and preparation of drug combination method thereof, the application in the anti-hepatitis B medicine of preparation of this compounds or its pharmaceutical composition.
In order to realize above-mentioned purpose of the present invention, the invention provides following technical scheme:
Herba Swertiae bimaculatae lactone H-K (1-4) shown in the structural formula (I),
Figure BSA00000385383000041
Pharmaceutical composition, wherein contain treat significant quantity formula (I) Herba Swertiae bimaculatae lactone H-K (1-4) any with the carrier that pharmaceutically can give and accept.
Described pharmaceutical composition is the pharmaceutical composition of treatment hepatitis B.
Any application in the medicine of preparation treatment human diseases or illness of formula (I) Herba Swertiae bimaculatae lactone H-K (1-4).
Above-mentioned application, described disease is a hepatitis B.
Contain the application of pharmaceutical composition in the medicine of preparation treatment human diseases or illness of formula (I) Herba Swertiae bimaculatae lactone H-K (1-4) any and the carrier that pharmaceutically can give and accept of treating significant quantity.
Above-mentioned application, described disease is a hepatitis B.
The method that the present invention prepares formula (I) Herba Swertiae bimaculatae lactone H-K (1-4) is to get Mile Swertia Herb (Swertia mileensis) herb, drying; Pulverize, use 90% and 50% alcohol reflux 3 times successively, each 2-3 hour; Merge alcohol extract, filter, decompression recycling ethanol is to there not being the alcohol flavor; This extracting solution is suspended in the aqueous solution, uses sherwood oil, ETHYLE ACETATE and n-butanol extraction successively, and ethyl acetate extraction part is adsorbed on the silica gel with the chloroform/methanol dissolving; Room temperature is placed and is volatilized, and grinds, through silica gel column chromatography; With chloroform/methanol (100: 1 → 80: 20) wash-out, be divided into ten parts, wherein first part uses silica gel column chromatography; Chloroform/acetone gradient elution with 95: 5 → 80: 20; Further pass through silica gel column chromatography then,, and obtain compound Herba Swertiae bimaculatae lactone H-K (1-4) through chloroform/methanol (1: 1) recrystallization with chloroform/methanol (95: 5) wash-out.
The method of pharmaceutical composition that preparation contains Herba Swertiae bimaculatae lactone H-K (1-4) is be raw material with compound Herba Swertiae bimaculatae lactone H-K (1-4), adding pharmaceutically acceptable carrier or vehicle.
When The compounds of this invention is used as medicine, can directly use, perhaps use with the form of pharmaceutical composition.This pharmaceutical composition contains 0.1-99%, is preferably the The compounds of this invention of 0.5-90%, and all the other are acceptable on the pharmacology, nontoxic and inert pharmaceutically acceptable carrier and/or vehicle to humans and animals.
Described pharmaceutical carrier or vehicle are one or more solids, 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 can be through injection (quiet notes, intramuscular injection) and oral two kinds of form administrations.
Embodiment:
In order to understand essence of the present invention better, below the pharmacological action result of formula of the present invention (I) Herba Swertiae bimaculatae lactone H-K (1-4) will be described with Test Example of the present invention, but not limit the present invention with this Test Example.
Test Example 1:
Herba Swertiae bimaculatae lactone H-K (I) is to the drug toxicity of Hep G2.2.15 cell and the restraining effect of HBV dna replication dna:
1 material and method
1.1 material: Herba Swertiae bimaculatae lactone H-K (I); Lamivudine (lamivudine) [GlaxoSmithKline PLC pharmacy (Suzhou) ltd, the accurate word H20030581 of traditional Chinese medicines]; Hep G2.2.15 cell (Guangzhou air hospital); High sugared DMEM (GIBCO); G418 (GIBICO); Foetal calf serum (Tianjin blood grinds institute); L-glutaminate (AMRESCO); Penicillium mould, Streptomycin sulphate [Zhongnuo Pharmaceutical (Shijiazhuang) Co., Ltd., Shiyao Group].
1.2 Hep G2.2.15 cell is cultivated with high sugared DMEM liquid, nutrient solution adds 10% foetal calf serum, 0.03%L-Stimulina, 100mg/L G418,10 5The blue or green enzyme of IU/L is plain, 100mg/L Streptomycin sulphate, 5%NaCO 3Transfer pH to 6.8-7.0.
1.3 instrument: ELIASA Bio-RAD 680 (U.S.); CO 2Incubator Thermo Forma 3310 (U.S.); Inverted biologic microscope XD-101 type (Nanjing); Quantitative real time PCR Instrument Mastercycler ep realplex type (German eppendorf company); Low-temperature and high-speed whizzer Centrifuge 5415D type (German eppendorf company).
1.4 experimentation: Hep G2.2.15 cell is with having added the 0.03%L-Stimulina, 100mg/L G418, the blue or green enzyme element of 100IU, high sugared DMEM (GIBICO) culture medium culturing of 100IU Streptomycin sulphate.Cell single cell suspension with after the trysinization is inoculated on the cell plate; Behind 48h, use instead and contain 2% foetal calf serum; The pastille substratum of 380 μ g/mL; It is four drug levels that every kind of medicine uses four times of dilutions of substratum, and cell control group and the acellular blank control group that does not add medicine is set simultaneously, and does the positive drug contrast with lamivudine.Measure the toxicity of medicine pair cell with mtt assay; Detect HBV DNA carrying capacity with fluorescence quantitative PCR method.
1.5 drug cell toxicity test.The cytotoxicity of the mtt assay detection of drugs of setting up according to Mosmann (Y.Nakajima, Y.Saton, M.Katsumata, K.Tsujiyama, Y.Ida, and J.Shoji, Phytochemistry 1994,36,119-127).Concrete grammar is: according to 5 * 10 5Individual every hole is inoculated in and changes supernatant behind the cell cultures 48h of 48 orifice plates is the pastille substratum.After continuing to cultivate 72h, adding concentration by the every hole of 0.3mL is the MTT of 0.4mg/mL, in 37 ℃ of 5%CO 2Abandon supernatant after hatching 4h, add the 0.3mL DMSO 99.8MIN. in every hole, hatch 10min in 37 ℃, measuring the absorbance of solution under 490nm on the ELIASA.Calculate the destruction percentage of medicine pair cell according to the result:
η Destroy=(A The cell control group-A Supply the test agent group)/(A The cell control group-A Blank control group) * 100.
1.6 medicine is to the influence of HBV DNA carrying capacity.Concrete grammar is: Hep G2.2.15 cell is by 5 * 10 5Individual every hole is inoculated in 24 porocyte plates, in 5%CO 2, be replaced by the pastille substratum behind the cultivation 48h in 37 ℃ of incubators, and every separated 48h changes cell conditioned medium with the pastille nutrient solution.Cell continued to cultivate after 8 days, and (TIANamp Gemomic DNA Kit, TIANGEN China) extract DNA to use blood/cell/tissue genome DNA extracting reagent kit.Method detection by quantitative HBV DNA carrying capacity with quantitative fluorescent PCR.1 μ L DNA sample is with 20 μ L2 * SYBR Green PCR Master Mix (Applied Biosystems; USA) and the special primer of 2 HBV as the pcr amplification system: preceding primer (5 ' GGA ACC TCT ATG TAT CCC TCC 3 '); Back primer (5 ' TCC GTC CGA AGG TTT GGT AC 3 '). amplification and detection Mastercycler Ep Realplex System quantitative PCR appearance (Eppendorf on probation; Masteraycler Eprealplex, German) 95 ℃ of preheatings 2 minutes, and by 95 ℃ 20 seconds; 58 ℃ 15 seconds, 72 ℃ of conditions of 20 seconds repeat 40 round-robin reactions.Calculate the inhibition percentage of medicine according to the result:
η Inhibition=(A The cell control group-A Supply the test agent group)/(A The cell control group-A Blank control group) * 100.
2. result: CC 50Be half cell-lethal concentration, according to destroying percentage η DestroyCalculate.IC 50For half virus inhibition concentration, according to suppressing percentage η InhibitionCalculate.Net result is with selectivity index (SI=CC 50/ IC 50) estimate.
CC 50, IC 50And the calculation formula of SI:
A=log (＞50% o'clock η Destroy/ η InhibitionDrug level)
B=log (＜50% o'clock η Destroy/ η InhibitionDrug level)
C＝|A-B|
CC 50=Anti the log [(η of 50-＜50% o'clock Destroy) * C/ (＞50% o'clock η Destroy-＜50% o'clock η Destroy)]+B
IC 50=the Antilog [(η of 50-＜50% o'clock Inhibition) * C/ (＞50% o'clock η Inhibition-＜50% o'clock η Inhibition)]+B
SI＝CC 50/IC 50
Concrete outcome is seen table 1:
Table 1 Herba Swertiae bimaculatae lactone H-K (1-4) is (I) in inhibition effect and the cytotoxicity (unit μ M) of Hep G2.2.15 cell to the HBV dna replication dna
Figure BSA00000385383000081
3, conclusion:
Experimental result shows that Herba Swertiae bimaculatae lactone H-K (1-4) has significant inhibitory effect external to the HBV dna replication dna, its IC 50Value and does not have obvious cytotoxicity between 1.53-5.34 μ M.
Below come further to illustrate preparation method of the present invention and medicine through embodiment and form, being situated between does not limit the present invention with this.
Embodiment 1:
Herba Swertiae bimaculatae lactone H-K (1-4) preparation (I):
The extraction separation of Herba Swertiae bimaculatae lactone H-K (1-4):
Gather the Mile Swertia Herb herb, its formal name used at school dries in the shade under the room temperature through being accredited as Swertia mileensis T.N.Ho et W.L.Shi; Pulverize, get 5.0kg dry powder, use 90% and 50% alcohol reflux 3 times successively; Each 2 hours, merge the ethanol extract, decompression recycling ethanol is to there not being the alcohol flavor.This extracting solution is suspended in the aqueous solution, uses sherwood oil, ETHYLE ACETATE and n-butanol extraction successively.Ethyl acetate extraction part is adsorbed on the silica gel with the chloroform/methanol dissolving, and room temperature is placed and volatilized solvent, grinds, and through silica gel column chromatography, with chloroform/methanol (100: 1 → 80: 20) wash-out, is divided into ten parts.Wherein first part continues to use silica gel column chromatography, with 95: 5 → 80: 20 chloroform/acetone gradient elution, further passes through the silicagel column purifying then; With chloroform/methanol (95: 5) wash-out, and, obtain Herba Swertiae bimaculatae lactone (swerilactone) H (1 respectively through chloroform/methanol (1: 1) recrystallization; 15.0mg), I (2; 19.0mg), J (3,20.0mg) and K (4,11.0mg).The structure of above compound is passed through 1H, 13C NMR, IR, UV, mass spectrum, and X-ray crystalline diffraction data are able to confirm.
The structured data of Herba Swertiae bimaculatae lactone H-K (1-4)
Fusing point is measured with the micro-fusing point appearance of XRC-1 type that appearance factory of section of Sichuan University produces, and TM is not revised; (Horiba, Tokyo Japan) measure by Jascomodel 1020 polarimeters in optically-active; Ir spectra (IR) adopts the KBr pressed disc method, by Bio-Rad FTS-135 type IR; UV spectrum is measured by UV-2401A type UVS; Nuclear magnetic resonance spectrum ( 1H-, 13C-NMR, DEPT) measure with DRX-500 type NMR spectrometer with superconducting magnet with Brucker AM-400 type and DRX-500 type NMR spectrometer with superconducting magnet mensuration, two dimensional NMR spectrum, with pyridine-d 5As solvent, mark in TMS (TMS) does; Mass spectrum (MS) is measured with VGAutospec-3000 type mass spectrograph; High resolution mass spectrum is measured with API Qstar Pulsar mass spectrograph; X-ray crystalline diffraction data use D/max-3B (Japanese company of science, Japan) and CAD4/PC (Enraf Noius & Enraf Noius company, Holland) x-ray diffractometer to measure respectively; Thin-layer chromatography silica gel, column chromatography silica gel (200-300 order) are available from Qingdao Makall Group Co., Ltd.;
Herba Swertiae bimaculatae lactone H (1)
Figure BSA00000385383000101
Molecular formula: C 30H 32O 11
Molecular weight: 568.56
Proterties: colourless prismatic crystal
mp：268-269℃
Optically-active
Figure BSA00000385383000102
(c=2.60mg cm -3, pyridine)
IR(KBr)v max：3456，1726，1701，1681，1642，1403，1278，1240，1107，1062，1041，1008，964，792cm -1。
UV/Vis(CHCl 3/MeOH＝1∶1，v/v)λ max(ε)：277(8989)nm。
ESIMS(-)m/z：603[M -+Cl]，567[M-H] -，341，297。
HRESIMS (-) m/z: experimental value 567.1876, calculated value 567.1866 (C 30H 31O 11, [M-H] -).
1H?NMR(C 5D 5N)δ：6.14(1H，br?s，H-7)，5.86(1H，br?s，H-11)，5.79(1H，brs，H-25)，5.23(1H，d，J＝2.6Hz，H-17)，5.10(1H，m，H-23)，4.96(1H，m，H-20a)，4.91(1H，d，J＝14.6Hz，H-20b)，4.61(1H，d，J＝?3.7Hz，H-13)，4.43(1H，m，H-3a)，4.42(1H，m，H-27a)，4.27(1H，m，H-3b)，3.91(1H，q，J＝6.2Hz，H-19)，3.89(1H，m，H-27b)，3.53(3H，s，H-33)，2.54(1H，m，H-16a)，2.52(1H，m，H-15)，2.29(1H，m，H-4a)，2.27(1H，m，H-28a)，2.16(1H，m，H-4b)，2.15(1H，m，H-14)，2.12(1H，m，H-28b)，1.47(3H，d，J＝6.8Hz，H-32)，1.46(1H，m，H-16b)，0.99(3H，d，J＝6.2Hz，H-31)。
13C?NMR(C 5D 5N)δ：172.4(s，C-30)，164.2(s，C-22)，161.0(s，C-1)，156.0(s，C-5)，141.0(s，C-9)，135.9(s，C-24)，132.4(s，C-8)，131.2(s，C-18)，130.1(s，C-29)，127.6(d，C-7)，126.4(s，C-10)，89.4(d，C-25)，76.3(d，C-19)，71.4(d，C-23)，68.5(t，C-20)，66.7(t，C-3)，66.2(d，C-13)，64.6(d，C-17)，63.3(d，C-11)，57.9(t，C-27)，54.3(s，C-12)，52.5(q，C-33)，47.7(s，C-6)，41.6(d，C-14)，36.1(d，C-15)，31.9(t，C-16)，27.2(t，C-4)，25.6(t，C-28)，18.2(q，C-31)，17.6(q，C-32)。
X-ray crystalline diffraction data: oblique system, spacer are p2 1
Figure BSA00000385383000111
Figure BSA00000385383000113
Figure BSA00000385383000114
α=90.00, β=96.91 (3), γ=90.00,
Figure BSA00000385383000115
Z=4, d=1.427g/cm 3, crystalline size 0.10 * 0.10 * 0.05nm 3, R 1=0.0374, wR 2=0.0710.
Herba Swertiae bimaculatae lactone I (2)
Molecular formula: C 29H 28O 10
Molecular weight: 536.53
Proterties: colourless prismatic crystal
mp：204-205℃
Optically-active
Figure BSA00000385383000121
(c=1.20mg cm -3, CHCl 3/ MeOH=1: 1, v/v)
IR(KBr)v max：3437，2925，1720，1672，1640，1462，1405，1376，1287，1247，1128，1100，1073，933，874cm -1。
UV/Vis(CHCl 3/MeOH＝1∶1，v/v)λ max(ε)：226(10896)，222(11110)，212(10977)nm。
ESIMS(+)m/z：559[M+Na] +。
HRESIMS (+) m/z: experimental value 559.1578, calculated value 559.1580 (C 29H 28O 10Na, [M+Na] +).
1H?NMR(C 5D 5N)δ：10.34(1H，s，H-25)，6.25(1H，s，H-7)，5.42(1H，s，H-11)，5.25(1H，s，H-17)，4.95(1H，d，J＝14.0Hz，H-20a)，4.89(1H，d，J＝14.0Hz，H-20b)，4.85(1H，m，H-23)，4.82(1H，m，H-27a)，4.64(1H，d，J＝3.2Hz，H-13)，4.48(1H，m，H-27b)，4.45(1H，m，H-3a)，4.40(1H，m，H-3b)，4.23(1H，d，J＝12.7Hz，H-14)，4.17(1H，m，H-19)，3.85(1H，m，H-28a)，3.67(1H，m，H-28b)，3.23(1H，m，H-15)，2.66(1H，m，H-4a)，2.48(1H，m，H-4b)，1.93(1H，m，H-16)，1.38(3H，d，J＝6.6Hz，H-32)，0.80(3H，d，J＝6.2Hz，H-31)。
13C?NMR(C 5D 5N)δ：189.5(d，C-25)，168.1(s，C-30)，163.7(s，C-22)，161.6(s，C-1)，159.0(s，C-5)，154.4(s，C-29)，144.3(s，C-9)，135.0(s，C-24)，131.3(s，C-8)，130.0(s，C-18)，128.4(d，C-7)，124.1(s，C-10)，71.3(d，C-19)，71.0(d，C-11)，68.9(d，C-23)，68.6(t，C-20)，67.6(t，C-27)，67.4(d，C-13)，66.8(t，C-3)，64.9(d，C-17)，53.5(s，C-12)，46.5(s，C-6)，37.4(d，C-14)，33.7(d，C-15)，30.8(t，C-16)，25.3(t，C-4)，24.8(t，C-28)，19.6(q，C-31)，18.8(q，C-32)。
X-ray crystalline diffraction data: oblique system, spacer are p2 1
Figure BSA00000385383000131
Figure BSA00000385383000132
Figure BSA00000385383000133
α=90, β=98.960 (3), γ=90,
Figure BSA00000385383000134
Figure BSA00000385383000135
Z=4, d=1.545g/cm 3, crystalline size 0.26 * 0.14 * 0.10nm 3, R 1=0.1069 (2391), wR 2=0.3491 (7831).
Herba Swertiae bimaculatae lactone J (3)
Figure BSA00000385383000136
Molecular formula: C 29H 28O 10
Molecular weight: 536.53
Proterties: colourless prismatic crystal
mp：266-267℃
Optically-active
Figure BSA00000385383000137
(c=2.52mg cm -3, CHCl 3/ MeOH=1: 1, v/v)
IR(KBr)v max：3434，1718，1690，1636，1460，1407，1266，1220，1182，1129，1108，1036，1023，1003，948，881cm -1。
UV/Vis(CHCl 3/MeOH＝1∶1，v/v)λ max(ε)：281(18499)，211(11673)，193(11756)nm。
ESIMS(+)m/z：1095[2M ++Na]，559[M+Na] +。
HRESIMS(+)m/z：found：559.1581[M ++Na]，calcd?for?559.1580(C 29H 28O 10Na，[M+Na] +)。
1H?NMR(C 5D 5N)δ：6.12(1H，s，H-7)，5.99(1H，br?s，H-28)，0.98(1H，d，J＝6.1Hz，H-31)，5.80(1H，s，H-11)，5.68(1H，br?s，H-25)，5.23?(1H，d，J＝14.6Hz，H-20a)，5.13(1H，d，J＝14.6Hz，H-20b)，5.04(1H，dd，J＝14.3，1.9Hz，H-27a)，4.92(1H，dd，J＝16.4，2.0Hz，H-27b)，4.50(1H，br?s，H-13)，4.44(1H，m，H-23)，4.33(1H，m，H-3a)，4.21(1H，m，H-3b)，3.93(1H，q，J＝6.3Hz，H-19)，2.66(1H，dd，J＝13.0，3.4Hz，H-14)，2.54(1H，br?s，H-24)，2.51(1H，m，H-15)，2.38(1H，m，H-16a)，5.14(1H，m，H-17)，2.31(1H，m，H-4a)，2.25(1H，m，H-4b)，1.35(1H，d，J＝6.6Hz，H-32)，1.19(1H，dd，J＝13.0，5.0Hz，H-16b)。
13C?NMR(C 5D 5N)δ：166.9(s，C-30)，164.0(s，C-22)，161.1(s，C-1)，156.2(s，C-5)，145.4(s，C-9)，132.6(s，C-8)，131.0(s，C-18)，129.4(s，C-29)，127.1(d，C-7)，123.5(s，C-10)，118.4(d，C-28)，95.6(d，C-25)，76.5(d，C-19)，73.0(d，C-23)，69.7(t，C-27)，68.7(t，C-20)，68.1(d，C-13)，66.7(t，C-3)，65.8(d，C-11)，64.7(d，C-17)，49.2(d，C-24)，48.0(s，C-6)，44.9(s，C-12)，42.9(d，C-14)，36.4(d，C-15)，31.0(t，C-16)，27.1(t，C-4)，18.4(q，C-31)，17.1(q，C-32)。
X-ray crystalline diffraction data: triclinic(crystalline)system, spacer are p2 1
Figure BSA00000385383000141
Figure BSA00000385383000143
α=70.965 (3), β=84.606 (3), γ=79.944 (3), Z=2, d=1.442g/cm 3, crystalline size 0.18 * 0.16 * 0.08nm 3, R 1=0.0925, wR 2=0.2123.
Herba Swertiae bimaculatae lactone K (4)
Molecular formula: C 29H 26O 9
Molecular weight: 518.51
Proterties: colourless prismatic crystal
mp：250-251℃
Optically-active
Figure BSA00000385383000151
(c=1.35mg cm -3, CHCl 3/ MeOH=1: 1, v/v)
IR(KBr)v max：3456，2958，2927，1713，1683，1645，1596，1468，1405，1281，1131，1103，1072，942cm -1。
UV/Vis(CHCl 3/MeOH＝1∶1，v/v)λ max(ε)：275(6337)，263(6122)，256(6058)，226(5641)，223(5666)，212(5505)，206(5407)nm。
ESIMS(+)m/z：541[M+Na] +。
HRESIMS (+) m/z: experimental value 541.1476, calculated value 541.1474 (C 29H 26O 9Na, [M+Na] +).
1H?NMR(C 5D 5N)δ：8.20(1H，d，J＝7.6Hz，H-26)，7.88(1H，d，J＝7.6Hz，H-24)，7.26(1H，dd，J＝7.6，7.6Hz，H-25)，6.26(1H，s，H-7)，6.40(1H，br?s，H-11)，5.74(1H，d，J＝9.8Hz，H-13)，5.28(1H，d，J＝14.1Hz，H-20a)，5.17(1H，d，J＝14.1Hz，H-20b)，4.88(1H，d，J＝3.5Hz，H-17)，4.49(1H，m，H-3)，4.46(1H，m，H-30b)，4.26(1H，m，H-30a)，3.39(1H，q，J＝6.1Hz，H-19)，3.25(1H，m，H-29)，2.95(1H，dd，J＝11.9，9.8Hz，H-14)，2.59(1H，m，H-4a)，2.39(1H，m，H-4b)，2.27(1H，m，H-15)，1.91(1H，m，H-16a)，1.01(1H，d，J＝6.1Hz，H-31)，0.65(1H，dd，J＝12.9，5.2Hz，H-16b)。
13C?NMR(C 5D 5N)δ：165.0(s，C-32)，162.7(s，C-22)，160.8(s，C-1)，155.9(s，C-5)，146.0(s，C-9)，138.9(s，C-23)，138.6(s，C-28)，133.2(d，C-24)，131.5(s，C-18)，131.0(d，C-26)，131.0(s，C-8)，127.8(d，C-25)，127.6(d，C-7)，126.6(s，C-27)，125.1(s，C-10)，88.5(d，C-11)，76.5(d，C-19)，69.0(t，C-20)，66.8(t，C-3)，66.8(t，C-30)，66.6(d，C-13)，64.2(d，?C-17)，47.5(s，C-6)，27.1(t，C-4)，42.8(d，C-14)，38.7(d，C-15)，32.8(t，C-16)，24.8(t，C-29)，18.3(q，C-31)。
X-ray crystalline diffraction data: oblique system, spacer are p2 1
Figure BSA00000385383000161
Figure BSA00000385383000162
α=90.00, β=106.921 (3), γ=90.00,
Figure BSA00000385383000164
Z=4, d=1.447g/cm 3, crystalline size 0.23 * 0.15 * 0.10nm 3, R 1=0.0656 (1877), wR 2=0.2035 (5631).
Embodiment 2:
Method by embodiment 1 makes Herba Swertiae bimaculatae lactone H-K (1-4) earlier, with after a spot of DMSO dissolving, adds the injection water by routine respectively, smart filter, and injection liquid is processed in the embedding sterilization.
Embodiment 3:
Method by embodiment 1 makes Herba Swertiae bimaculatae lactone H-K (1-4) earlier, with after a spot of DMSO dissolving, it is dissolved in the sterile water for injection respectively; Stirring makes dissolving, filters with aseptic suction funnel, aseptic more smart filter; Be sub-packed in the ampoule, aseptic sealing by fusing gets powder injection behind the frozen drying.
Embodiment 4:
With separate of the Herba Swertiae bimaculatae lactone H-K (1-4) that obtains, be that 9: 1 ratio adds vehicle in itself and vehicle weight ratio respectively, process pulvis.
Embodiment 5:
Method by embodiment 1 makes Herba Swertiae bimaculatae lactone H-K (1-4) earlier, is that 5: 1 ratio adds vehicle, pelletizing press sheet in itself and vehicle weight ratio respectively.
Embodiment 6:
Method by embodiment 1 makes Herba Swertiae bimaculatae lactone H-K (1-4) earlier, processes oral liquid by conventional oral liquid method for making respectively.
Embodiment 7:
Method by embodiment 1 makes Herba Swertiae bimaculatae lactone H-K (1-4) earlier, is that 5: 1 ratio adds vehicle in itself and vehicle weight ratio respectively, processes capsule.
Embodiment 8:
Method by embodiment 1 makes Herba Swertiae bimaculatae lactone H-K (1-4) earlier, is that 3: 1 ratio adds vehicle in itself and vehicle weight ratio respectively, processes capsule.

Claims (9)

1. the Herba Swertiae bimaculatae lactone H-K (1-4) shown in the structural formula (I),
2. pharmaceutical composition, wherein contain the said formula of the claim 1 of treating significant quantity (I) Herba Swertiae bimaculatae lactone H-K (1-4) any with pharmaceutically acceptable carrier.
3. pharmaceutical composition as claimed in claim 2 is the pharmaceutical composition of treatment hepatitis B.
4. any application in the medicine of preparation treatment human diseases or illness of the said formula of claim 1 (I) Herba Swertiae bimaculatae lactone H-K (1-4).
5. application as claimed in claim 4, described disease is a hepatitis B.
6. the application of the said pharmaceutical composition of claim 2 in the medicine of preparation treatment human diseases or illness.
7. application as claimed in claim 6, described disease is a hepatitis B.
8. the method for preparing the said formula of claim 1 (I) Herba Swertiae bimaculatae lactone H-K (1-4) is characterized in that getting Mile Swertia Herb (Swertia mileensis) herb, drying; Pulverize; Use 90% and 50% alcohol reflux 3 times successively, each 2-3 hour, merge the ethanol extract; Filter, being evaporated to does not have the alcohol flavor; This extracting solution is suspended in the aqueous solution; Use sherwood oil, ETHYLE ACETATE and n-butanol extraction successively, ethyl acetate extraction part is adsorbed on the silica gel with the chloroform-methanol dissolving, and room temperature is placed and volatilized solvent; Grind; Through silica gel column chromatography,, obtain ten parts with 100: 1 → 80: 20 chloroform/methanol wash-out; Wherein first part continues to use silica gel column chromatography; Chloroform/acetone gradient elution with 95: 5 → 80: 20; Further pass through silica gel column chromatography then, with 95: 5 chloroform/methanol wash-out, and 1: 1 chloroform/methanol recrystallization of warp obtained compound Herba Swertiae bimaculatae lactone H-K (1-4).
9. the method for preparing the described pharmaceutical composition of claim 2, compound Herba Swertiae bimaculatae lactone H-K (1-4) adds pharmaceutically acceptable carrier or vehicle respectively.
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CN104497004A (en) * 2014-12-30 2015-04-08 云南中医学院 Method for preparing swertia lactone I-K
CN104497005A (en) * 2014-12-30 2015-04-08 云南中医学院 Swertiamarin thermal conversion product, as well as preparation method, preparation and application thereof

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CN104478893A (en) * 2014-12-30 2015-04-01 云南中医学院 Lactone compound of false Chinese swertia herb, as well as preparation method, preparation and application of lactone compound
CN104497004A (en) * 2014-12-30 2015-04-08 云南中医学院 Method for preparing swertia lactone I-K
CN104497005A (en) * 2014-12-30 2015-04-08 云南中医学院 Swertiamarin thermal conversion product, as well as preparation method, preparation and application thereof
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