CN114057764B - Linderane type dimeric sesquiterpene with anti-inflammatory activity and preparation method and application thereof - Google Patents

Linderane type dimeric sesquiterpene with anti-inflammatory activity and preparation method and application thereof Download PDF

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CN114057764B
CN114057764B CN202111486837.8A CN202111486837A CN114057764B CN 114057764 B CN114057764 B CN 114057764B CN 202111486837 A CN202111486837 A CN 202111486837A CN 114057764 B CN114057764 B CN 114057764B
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column chromatography
methanol
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linderane
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倪伟
严欢
李萍
肖龙高
刘海洋
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Kunming Institute of Botany of CAS
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
    • C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
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Abstract

The invention belongs to the field of phytochemistry and medicines, relates to linderane type dimeric sesquiterpene sarcglabralides A-C with anti-inflammatory activity, a preparation method and application thereof, a pharmaceutical composition containing sarcglabralides A-C as an active pharmaceutical ingredient and pharmaceutical application of sarcglabralides A-C. The linderane dimeric sesquiterpene sarcglabralides A-C has better anti-inflammatory activity and can be used for treating inflammatory diseases.

Description

Linderane type dimeric sesquiterpene with anti-inflammatory activity and preparation method and application thereof
Technical Field
The invention belongs to the technical field of phytochemistry and medicines, and particularly relates to three linderane type dimeric sesquiterpenes sarcglabralides A-C with anti-inflammatory activity, a pharmaceutical composition containing any one or two or three of the linderane type dimeric sesquiterpenes, a preparation method of a compound sarcglabralides A-C and application of the compound sarcglabralides A-C in preparation of anti-inflammatory medicines.
Background
Inflammation is an active defensive response of living tissue with the vascular system to endogenous or exogenous stimuli. Under normal physiological conditions, moderately controlled inflammation can help the body clear and repair damage. Under pathological conditions, uncontrolled and excessive inflammation can in turn lead to damage to tissues and organs. Chronic inflammation, which persists for a long period of time, presents a number of health hazards, such as: inducing cancer occurrence, helicobacter pylori infectious inflammation, EB virus infectious inflammation, and hepatitis virus inflammation, respectively associated with gastric cancer, nasopharyngeal carcinoma and liver cancer; cardiovascular health is compromised-chronic inflammation caused by fat plaques in blood vessels can form thrombi which ultimately lead to pulmonary arterial hypertension, chronic obstructive pulmonary disease, emphysema, heart disease and the like; inducing Alzheimer's disease-with age, the inflammatory response causes a disturbance of the regulatory mechanisms of the immune system, increasing the occurrence of Alzheimer's disease; in addition, chronic excessive inflammatory reactions can also cause diabetes and its complications, parkinson's disease, macular degeneration, osteoarthritis, and the like. With the increasing speed of urbanization and aging processes and changes in lifestyle, inflammatory diseases have become one of the important threats affecting the health of the global population, are spreading worldwide and have evolved into important public health problems seriously jeopardizing human health and sustainable development of socioeconomic.
The inflammatory response involves a series of physiological and pathological changes: such as increased capillary permeability of damaged tissue, release of arachidonic acid metabolites and platelet-activating factors, release of inflammatory factors (IL-1, IL-6, TNF-alpha, histamine, 5-hydroxytryptamine), etc., and these physiological and pathological changes constitute an inflammatory response network. NO plays a key regulatory role in the inflammatory response, particularly in the occurrence of inflammatory responses and signaling. NO is a sequence of regulatory actions by acting on autogenous cells or diffusing to neighboring cells and binding to its target receptor (transcription factor, protein kinase, etc.) after the production of L-arginine is catalyzed by Nitric Oxide Synthase (NOS). In the immune inflammation process, NO in the body can activate NF- κB under the high concentration state, induce the production of pro-inflammatory cytokines TNF-alpha, IL-1β and the like, and the production of IL-1 and TNF-alpha can activate iNOS (inducible nitric oxide synthase) to promote the body to produce more NO, so that the secretion of cytokines and the self-generated expression of NO can be sustained, and the inflammatory reaction is more durable and more severe. Thus, inhibiting the production of NO in an inflammatory response can result in an improvement in inflammatory symptoms.
Currently, the treatment of inflammation is mainly a chemical synthesis of steroids (glucocorticoids: such as dexamethasone, prednisone, hydrocortisone, etc.) and non-steroids (NSAIDS: such as aspirin, ibuprofen, indomethacin, etc.), which have various side effects, such as: gastrointestinal reactions, cardiotoxicity, hepatotoxicity, nephrotoxicity, immune dysfunction, and the like. Therefore, the search for new anti-inflammatory lead compounds is continued, and research and development of anti-inflammatory drugs with good curative effects and small side effects are necessary and urgent.
The natural products undergo a long and lengthy evolution process in nature, the novelty of chemical structures and the diversity of biological activities are one of important resources for drug development, and the search for lead compounds with anti-inflammatory activity from natural products has great potential.
The invention comprises the following steps:
therefore, the invention aims at three new linderane type dimeric sesquiterpene natural products, namely the compounds of the Sarcabralide A, the Sarcabralide B and the Sarcabralide C, and the application of the three compounds as active ingredients in preparing anti-inflammatory medicaments, thereby providing natural products with good curative effect and small side effect for the treatment of inflammatory diseases.
The object of the invention can be achieved by the following technical scheme.
In a first aspect, the present invention provides linderane dimeric sesquiterpenes, namely compounds sarcglabralides A, B and C, having the structural formulae shown below in formulas I, ii and iii, respectively:
in a second aspect, the present invention provides a pharmaceutical composition for the treatment of inflammatory diseases comprising as active pharmaceutical ingredient a linderane dimeric sesquiterpene compound sarcglabralides A, B or C or a combination thereof, or a pharmaceutically acceptable derivative thereof, together with pharmaceutically acceptable carriers, adjuvants and excipients.
In a third aspect, the invention relates to the use of a linderane dimeric sesquiterpene compound sarcglabralides A, B or C or a pharmaceutically acceptable derivative thereof for the manufacture of a medicament for the treatment of inflammatory diseases.
In a fourth aspect, the present invention provides a process for preparing a linderane dimeric sesquiterpene compound sarcglabralides A, B or C, comprising the steps of:
1) Drying and pulverizing herba Pileae Scriptae (Sarcandra) plant material, extracting with organic solvent, and desolventizing to obtain herba Pileae Scriptae plant material extract;
2) And (3) sequentially adopting column chromatography, medium-pressure separation gel and semi-preparation high-pressure liquid chromatography to obtain a compound sarcglabralides A-C.
In a preferred embodiment, the organic solvent comprises at least one of petroleum ether, chloroform, methylene chloride, ethyl acetate, acetone, ethanol, methanol, n-butanol, acetonitrile, and formic acid.
Compared with the prior art, the invention has obvious beneficial effects. In particular, the linderane type dimeric sesquiterpene sarcglabralides A-C has remarkable anti-inflammatory activity and can be used for treating inflammatory diseases.
Drawings
FIG. 1 shows the chemical structural formula of the linderane dimeric sesquiterpene compound sarcglabralides A-C.
FIG. 2 is a graph showing the results of ECD (electronic circular dichroism) calculation of absolute configuration of the linderane dimeric sesquiterpene sarcglabralides A-C compound of the invention.
Detailed Description
The present invention will be described more fully hereinafter in order to facilitate an understanding of the present invention, and preferred embodiments of the present invention are set forth. This invention may, however, be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the scope of the invention. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
According to a first aspect, the present invention provides linderane dimeric sesquiterpene compounds sarcglabralides A, B and C having the structural formulae shown in formulae I, ii and iii below, respectively:
according to a second aspect of the present invention there is provided a pharmaceutical composition for use in the treatment of inflammatory diseases comprising as an active pharmaceutical ingredient a linderane dimeric sesquiterpene compound sarcglabralides A, B or C or a pharmaceutically acceptable derivative thereof, together with a pharmaceutically acceptable carrier, adjuvant or excipient.
In embodiments of the pharmaceutical compositions of the present invention, inflammatory diseases include, but are not limited to: skin inflammation, rheumatic diseases, cardiovascular diseases, autoimmune diseases, etc.
In an embodiment of the pharmaceutical composition of the present invention, the linderane-type dimeric sesquiterpenes as the active ingredient may be any one, any two or three of the compounds sarcglabralides A-C.
It will be appreciated by those skilled in the art that pharmaceutically acceptable derivatives of the compounds sarcglabralides A-C of the present invention, such as salts, solvates or hydrates of the compounds sarcglabralides A-C, may also be used in the pharmaceutical compositions of the present invention.
The pharmaceutically acceptable salt may be, for example, a pharmaceutically acceptable addition base salt.
Pharmaceutically acceptable addition salts include salts derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Salts derived from pharmaceutically acceptable non-toxic organic bases include salts of primary, secondary and tertiary amines, including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine and triethanolamine.
Thus, in embodiments of the pharmaceutical compositions of the present invention, the active pharmaceutical ingredient may be any one of compounds sarcglabralides A-C, mixtures of any two or three, or a base addition salt, solvate, hydrate or the like derivative thereof.
Suitable pharmaceutical excipients are well known to those skilled in the art. Pharmaceutically acceptable carriers or excipients are one or more solid, semi-solid and liquid diluents, fillers and pharmaceutical product adjuvants including, but not limited to, fillers (diluents), lubricants (glidants or anti-adherents), dispersants, wetting agents, binders, solubilizing agents, antioxidants, bacteriostats, emulsifiers, disintegrants and the like. The binder comprises syrup, acacia, gelatin, sorbitol, tragacanth, cellulose and its derivatives (such as microcrystalline cellulose, sodium carboxymethylcellulose, ethylcellulose or hydroxypropyl methylcellulose), gelatin slurry, syrup, starch slurry or polyvinylpyrrolidone; the filler comprises lactose, sugar powder, dextrin, starch and its derivatives, cellulose and its derivatives, inorganic calcium salt (such as calcium sulfate, calcium phosphate, calcium hydrogen phosphate, precipitated calcium carbonate, etc.), sorbitol or glycine, etc.; the lubricant comprises aerosil, magnesium stearate, talcum powder, aluminum hydroxide, boric acid, hydrogenated vegetable oil, polyethylene glycol and the like; disintegrants include starch and its derivatives (e.g., sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch, etc.), polyvinylpyrrolidone, microcrystalline cellulose, etc.; the wetting agent comprises sodium dodecyl sulfate, water or alcohol, etc.; the antioxidant comprises sodium sulfite, sodium bisulphite, sodium metabisulfite, dibutyl benzoic acid and the like; the bacteriostat comprises 0.5% phenol, 0.3% cresol, 0.5% chlorobutanol and the like; the emulsifier comprises polysorbate-80, sorbitan without acid, lecithin, soybean lecithin, etc.; the solubilizer comprises Tween-80, bile, glycerol, etc.
When the linderane dimeric sesquiterpenes of the present invention are used as medicaments, they may be administered directly or in the form of pharmaceutical compositions. In the pharmaceutical composition of the present invention, the pharmaceutical composition may contain sarcglabralides A, B or C in an amount of 0.1 to 99%, preferably 0.5 to 90% by weight of the total pharmaceutical composition.
An "effective amount" of a linderane dimeric sesquiterpene compound Sarcglabralides A, B or C or a pharmaceutically acceptable derivative thereof, e.g. a base addition salt, refers to an amount sufficient to achieve the desired biological effect. It will be appreciated that the effective dose will depend on the age, sex, health condition and weight of the recipient. Typically, the effective amount is determined by the person administering the treatment, e.g., a treating physician.
The pharmaceutical composition of the present invention may be administered in the form of a unit weight dose. All pharmaceutical compositions containing the linderane dimeric sesquiterpene sarcglabralides A-C or pharmaceutically acceptable derivatives thereof such as base addition salts as active ingredients are prepared into various dosage forms such as liquid preparations (injections, suspensions, emulsions, solutions, syrups and the like), solid preparations (tablets, capsules, granules, medicinal granules and the like), sprays, aerosols and the like by adopting methods accepted in the pharmaceutical and food fields. The pharmaceutical composition of the present invention can be used for the treatment of inflammatory diseases by injection (intravenous injection, intravenous drip, intramuscular injection, intraperitoneal injection, subcutaneous injection), oral administration, sublingual administration, mucosal dialysis and other administration routes.
According to a third aspect, the present invention provides the use of a linderane dimeric sesquiterpene compound sarcglabralides A-C or a pharmaceutically acceptable derivative thereof according to the invention in the manufacture of a medicament for the treatment of inflammatory diseases.
In embodiments of the invention, inflammatory diseases include, but are not limited to: skin inflammation, rheumatic diseases, cardiovascular diseases, autoimmune diseases, etc.
According to a fourth aspect, the present invention provides a process for preparing a linderane dimeric sesquiterpene compound sarcglabralides A-C, comprising the steps of:
1) Drying and pulverizing plant material of Sarcandra (Sarcandra), leaching with organic solvent, and desolventizing to obtain Sarcandra plant extract;
2) And (3) sequentially adopting column chromatography, medium pressure chromatography to separate gel and semi-preparative high performance liquid chromatography to obtain sarcglabralides A-C.
In particular embodiments, the sarcandra plant material may be various parts of a sarcandra plant, such as roots, stems, branches, leaves or fruits, or whole plants. In embodiments of the invention, the sarcandra plant material may be parts (e.g., roots, stems, branches, leaves, or fruits) or whole plants of sarcandra (s.glabra), sarcandra (s.hainanensis) plants. In a preferred embodiment of the invention, the sarcandra plant material is from sarcandra (s.glabra).
In an embodiment of the present invention, the organic solvent used for the extraction in step 1) includes at least one of petroleum ether, chloroform, methylene chloride, ethyl acetate, acetone, ethanol, methanol, n-butanol, acetonitrile, and formic acid, and preferably, the organic solvent is methanol, ethyl acetate, or 95% ethanol.
In the embodiment of the present invention, the extraction method may be organic solvent cold leaching extraction, heating reflux extraction, ultrasonic extraction, or the like.
In an embodiment of the invention, the sarcandra material is extracted with the organic solvent at least 1 time, for example 2, 3, 4 times, preferably 3 times.
In embodiments of the invention, the volume ratio of organic solvent to dried sarcandra plant material is from 1:1 to 5:1, e.g., 1:1, 2:1, 3:1, 4:1, 5:1, and ratios between any two of the foregoing, e.g., 1.5:1, 2.5:1, 3.5:1, 4.5:1, etc., preferably 3:1.
In a specific embodiment of the present invention, step 2) specifically comprises:
in particular embodiments of the present invention, in step 2), the column chromatography includes normal phase silica Gel column chromatography, reverse phase silica Gel column chromatography (e.g., RP-18), gel column chromatography (e.g., sephadex LH-20), medium pressure chromatographic separation gels (e.g., MCI Gel CHP 20P), and preparative or semi-preparative High Performance Liquid Chromatography (HPLC).
In a specific embodiment of the present invention, in step 2), the eluent used for column chromatography comprises one or a combination of any two of petroleum ether, chloroform, methylene chloride, ethyl acetate, acetone, ethanol, methanol, n-butanol, acetonitrile, water and formic acid; the normal phase silica gel column chromatography adopts petroleum ether and ethyl acetate to carry out gradient elution according to the volume ratio from 1:0 to 0:1, such as 1:0, 50:1, 25:1, 10:1, 5:1 and 0:1; or normal phase silica gel column chromatography is eluted with chloroform and methanol in a volume ratio from 100:1 to 1:1, for example 100:1, 50:1, 30:1, 10:1, 1:1; reversed-phase RP-18 column chromatography adopts methanol and water to carry out gradient elution according to the volume ratio of 2:8 to 1:0; gel column chromatography, e.g., sephadex LH-20 gel column chromatography, eluting with methanol, chloroform-methanol (v: v 1:1); the medium pressure chromatographic separation Gel (MCI Gel) is eluted with methanol and water in the volume ratio of 2:8 to 1:0; preparative or semi-preparative HPLC eluting with a gradient from 2:8 to 6:4 volume ratio of acetonitrile and water.
Preferred embodiments of the present invention will be described in detail with reference to examples. It should be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and substitutions may be made by those skilled in the art without departing from the spirit and scope of the invention, all such modifications and substitutions being within the scope of the invention as set forth in the appended claims.
The experimental methods used in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Measuring by an Agilent UPLC/Q-Tof liquid chromatography-mass spectrometer; ultraviolet (UV) spectroscopy in methanol with a Shimazu UV-2401PC ultraviolet spectrometer; HPLC analysis and preparation were performed using an Agilent type 1260 or 1100 high performance liquid chromatograph, with a column of AgilentZORBAX-SB-C18 chromatography in the examples described below, 1 H, 13 c NMR and 2D NMR spectra were measured on a Bruker DRX-500 NMR spectrometer; electrospray ionization mass spectrometry (ESI-MS) was determined by a Waters XenvoTQ-S triple quadrupole tandem mass spectrometer or a Bruker HTC/Esquire liquid-ion trap chromatography mass spectrometer; high resolution mass spectrometry (HR-ESI-MS) was performed on columns (5 μm; 4.6X100 mm) or Agilent ZORBAX-SB-C18 chromatography columns (5 μm; 9.4X1250 mm); the normal phase silica gel (200-300 meshes) for column chromatography and the thin layer chromatography plate are all products of Qingdao ocean chemical plants; thin layer chromatography by 10% FeCl 3 -observation of the spots with an ethanol solution; sephadex LH-20 is a product of GE Healthcare; the reversed phase material Rp-8 and Rp-18 are products of Merck company. MCI Gel CHP20P is a Mitsubishi chemical product of Japan.
Example 1: process for preparing compound sarcglabralides A-C
Crushing sarcandra glabra (S.glabra) root, extracting with ethyl acetate at room temperature in the volume ratio of organic solvent to plant material of 3:1 for 4 times (2 days/time), mixing the extractive solutions, and distilling under reduced pressure to obtain pasty extract. The pasty extract was subjected to silica gel column chromatography and eluted with a petroleum ether-ethyl acetate gradient (40:1.fwdarw.1:1, v/v) to give 6 fractions (Fr.1-Fr.6). Subjecting the collected Fr.4 fraction to RP-18 reverse phase column chromatography, eluting with methanol-water gradient (2:8→1:0, v/v) to obtain 6 components Fr.4.1-Fr.4.6.Fr.4.3 gel column chromatography (Sephadex LH-20) with chloroform: methanol (1:1, v/v) is used as an eluent, and then the sarcoglabra A and the sarcoglabra B can be prepared by a semi-preparative High Performance Liquid Chromatography (HPLC) with acetonitrile-water as a mobile phase and adopting an isocratic elution method (the concentration of acetonitrile is 45%, v/v). Fr.4.5 was eluted with chloroform-methanol (30:1, v/v) using normal phase silica gel column chromatography to give sarcoglabra C.
Physical constants and spectral data for sarcoglabra a: yellow gum; UV(MeOH)λ max (logε)216(2.93)nm；CD(MeOH)λ max (Δε) 225(3.50)nm；IR(KBr)ν max 3440,2925,2853,1720,1632,1454,1384,1263, 1128cm -1 ； 1 H NMR(CDCl 3 ,600MHz)δ2.06(1H,dt,J＝9.1,5.2Hz,H-1), 0.83(1H,m,H-2α),1.72(1H,m,H-2β),1.67(1H,o,H-3),4.26(1H,s,H-9), 1.34(3H,s,H 3 -13),1.06(3H,s,H 3 -14),1.58(1H,m,H-15α),2.64(1H,dd,J ＝14.6,3.7Hz,H-15β),1.67(1H,o,H-1′),0.64(1H,td,J＝8.9,5.5Hz, H-2′α),1.17(1H,m,H-2′β),1.70(1H,o,H-3′),1.82(1H,o,H-5′),1.91(1H, d,J＝12.8Hz,H-6′α),1.80(1H,d,J＝12.8Hz,H-6′β),2.83(1H,dd,J＝13.5,3.8Hz,H-9′),6.55(1H,s,H-13′a),5.70(1H,s,H-13′b),0.91(3H,s, H 3 -14′),4.07(1H,d,J＝11.0Hz,H-15′a),3.93(1H,d,J＝11.0Hz,H-15′b), 6.88(1H,q,J＝7.1Hz,H-3″),1.82(3H,d,J＝7.1Hz,H 3 -4″),1.85(3H,s, H 3 -5″),6.12(1H,s,H-1″′),3.47(3H,s,MeO-1″′),3.77(3H,s,MeO-12)； 13 C NMR(CDCl 3 ,150MHz)δ31.9(C-1,d),9.5(C-2,t),27.2(C-3,d),92.6 (C-4,s),90.2(C-5,s),148.1(C-6,s),143.1(C-7,s),198.1(C-8,s),81.2(C-9,d),54.8(C-10,s),63.3(C-11,s),173.0(C-12,s),25.6(C-13,q),14.3(C-14, q),33.2(C-15,t),27.1(C-1′,d),10.8(C-2′,t),29.3(C-3′,d),78.7(C-4′,s), 53.1(C-5′,d),33.2(C-6′,t),57.7(C-7′,s),95.9(C-8′,s),50.0(C-9′,d),41.5 (C-10′,s),143.5(C-11′,s),167.6,(C-12′,s),126.6(C-13′,t),23.8(C-14′,q), 69.4(C-15′,t),168.0(C-1″,s),128.4(C-2″,s),138.2(C-3″,d),14.6(C-4″, q),12.3(C-5″,q),121.2(C-1″′,d),54.1(MeO-1″′,q),53.3(MeO-12,q)。 HRESIMS m/z 727.2537[M+Cl] - (calcd.for C 38 H 44 O 12 Cl,727.2527)。
physical constants and spectral data for sarcoglabra B: yellow gum; UV(MeOH)λ max (logε)220(3.13)nm；CD(MeOH)λ max (Δε) 223(14.8),333(–1.04)nm；IR(KBr)ν max 3446,2928,2854,1714,1643, 1441,1381,1263,1132cm -1 ； 1 H NMR(CDCl 3 ,500MHz)δ2.03(1H,dt,J＝ 8.6,5.0Hz,H-1),0.73(1H,m,H-2α),0.81(1H,m,H-2β),2.28(1H,ddd,J＝8.6,5.9,3.6Hz,H-3),4.23(1H,s,H-9),1.33(3H,s,H 3 -13),1.02(3H,s, H 3 -14),1.40(1H,t,J＝14.3Hz,H-15α),3.31(1H,dd,J＝14.6,3.1Hz, H-15β),1.58(1H,m,H-1′),0.61(1H,td,J＝8.9,5.5Hz,H-2′α),1.14(1H,m,H-2′β),1.69(1H,m,H-3′),1.76(1H,s,H-5′),1.92(1H,d,J＝11.5Hz, H-6′α),1.75(1H,d,J＝11.5Hz,H-6′β),2.42(1H,dd,J＝13.9,3.0Hz,H-9′), 6.56(1H,s,H-13′a),5.72(1H,s,H-13′b),0.86(3H,s,H 3 -14′),4.06(1H,d,J ＝11.0Hz,H-15′a),3.89(1H,d,J＝11.0Hz,H-15′b),6.87(1H,q,J＝7.1Hz, H-3″),1.81(3H,d,J＝7.1Hz,H 3 -4″),1.84(3H,s,H 3 -5″),2.23(1H,q, H-2″′),3.77(3H,s,MeO-12)；3.18(1H,s,5-OH),3.42(1H,d,J＝4.0Hz, 9-OH)； 13 C NMR(CDCl 3 ,125MHz)δ29.9(C-1,d),7.7(C-2,t),27.1(C-3, d),89.4(C-4,s),78.6(C-5,s),150.3(C-6,s),141.3(C-7,s),198.1(C-8,s), 80.9(C-9,d),54.6(C-10,s),63.3(C-11,s),173.0(C-12,s),25.7(C-13,q),13.7(C-14,q),31.7(C-15,t),27.1(C-1′,d),10.9(C-2′,t),29.3(C-3′,d),78.7 (C-4′,s),52.8(C-5′,d),32.9(C-6′,t),57.4(C-7′,s),96.7(C-8′,s),50.1(C-9′, d),41.3(C-10′,s),142.9(C-11′,s),167.6,(C-12′,s),127.1(C-13′,t),24.1 (C-14′,q),69.4(C-15′,t),168.0(C-1″,s),128.4(C-2″,d),138.2(C-3″,d), 14.6(C-4″,q),12.3(C-5″,q),169.3(C-1″′,s),22.2(Me-2″′,q),53.3 (MeO-12,q)。HRESIMS m/z 715.2723[M+Na] + (calcd.for C 38 H 44 O 12 Na, 715.2725)。
physical constants and spectral data for sarcoglabra C: yellow gum; UV(MeOH)λ max (logε)214(3.24)nm；CD(MeOH)λ max (Δε) 260(5.93),212(–9.53),343(–1.58)nm；IR(KBr)ν max 3440,2955,2924, 1738,1636,1437,1382,1241,1126cm -1 ； 1 H NMR(CDCl 3 ,600MHz)δ2.05 (1H,m,H-1),0.98(1H,m,H-2α),0.28(1H,m,H-2β),1.83(1H,o,H-3), 3.91(1H,s,H-6),3.93(1H,s,H-9),1.88(3H,s,H 3 -13),1.01(3H,s,H 3 -14), 2.59(1H,dt,J＝16.5,5.1Hz,H-15α),2.75(1H,d,J＝16.5Hz,H-15β),1.58(1H,o,H-1′),0.72(1H,td,J＝8.7,6.0Hz,H-2′α),1.28(1H,m,H-2′β),1.43 (1H,td,J＝8.6,3.6Hz,H-3′),1.75(1H,dd,J＝13.8,5.8Hz,H-5′),2.41(1H,dd,J＝18.4,5.8Hz,H-6′α),2.72(1H,dd,J＝18.4,13.8Hz,H-6′β),1.83 (1H,o,H-9′),4.81(1H,s,H-13′a),4.81(1H,s,H-13′b),0.86(3H,s,H 3 -14′), 4.06(1H,d,J＝11.6Hz,H-15′a),3.75(1H,d,J＝11.6Hz,H-15′b),2.13 (3H,s,H 3 -2″),2.09(3H,s,H-2″′),3.78(3H,s,MeO-12)； 13 C NMR(CDCl 3 , 150MHz)δ25.5(C-1,d),16.0(C-2,t),24.8(C-3,d),142.4(C-4,s),131.7(C-5,s),40.9(C-6,d),131.4(C-7,s),200.5(C-8,s),80.4(C-9,d),51.3 (C-10,s),148.0(C-11,s),170.6(C-12,s),20.6(C-13,q),15.3(C-14,q),25.4(C-15,t),25.8(C-1′,d),12.0(C-2′,t),28.2(C-3′,d),77.2(C-4′,s),60.7 (C-5′,d),23.0(C-6′,t),171.5(C-7′,s),93.3(C-8′,s),55.9(C-9′,d),44.8 (C-10′,s),123.7(C-11′,s),172.3,(C-12′,s),55.2(C-13′,t),26.5(C-14′,q), 71.6(C-15′,t),170.7(C-1″′,s),20.6(C-2″′,q),52.8(MeO-12,q)。HRESIMS m/z 659.2465[M+Na] + (calcd.for C 35 H 40 O 11 Na,659.2463)。
example 2: preparation method II of compound Sarcabralides sA-C
Extracting herba Pileae Scriptae (S.glabra) with 95% ethanol under reflux for 3 times at a volume ratio of organic solvent to plant material of 2:1 (each time for 2 hr), mixing extractive solutions, vacuum distilling to recover small volume (ethanol concentration of 25% -30%), purifying with LS-700B macroporous resin column, eluting with 5 column volumes of distilled water to remove sugar, eluting with 75% ethanol, and recovering ethanol solvent to obtain pasty extract. The paste was chromatographed on an MCI column, eluting with methanol-water gradients (3:7, 5:5, 8:2, v/v), 4-6 column volumes per gradient, to give 3 fractions (Fr.1-Fr.3). Subjecting the collected Fr.2 part to normal phase silica gel column chromatography (the mass ratio of the silica gel to the sample is 20:1), and gradient eluting with petroleum ether-ethyl acetate as an eluent (3-5 column volumes per gradient) to obtain six parts: fr.2.1 (petroleum ether: ethyl acetate=1:0, v/v), fr.2.2 (petroleum ether: ethyl acetate=50:1, v/v), fr.2.3 (petroleum ether: ethyl acetate=25:1, v/v), fr.2.4 (petroleum ether: ethyl acetate=10:1, v/v), fr.2.5 (petroleum ether: ethyl acetate=5:1, v/v); fr.2.6 (petroleum ether: ethyl acetate=0:1, v/v); fr.2.5 gel column chromatography (Sephadex LH-20) with chloroform: methanol (1:1, v/v) is used as an eluent, and then the sarcoglabra A, the sarcoglabra B and the sarcoglabra C are prepared by a semi-preparative High Performance Liquid Chromatography (HPLC) with acetonitrile-water as a mobile phase and a gradient elution method (the concentration of acetonitrile is 20% -60%, v/v).
Example 3: preparation method three of compound Sarcabralides A-C
Branch and leaf (dried) of sarcandra glabra (S.glabra), pulverizing, extracting with methanol at room temperature at a volume ratio of organic solvent to plant material of 5:1 for 3 times (2 days/time), mixing the extractive solutions, and distilling under reduced pressure to obtain pasty extract. Mixing the extractive solutions, distilling under reduced pressure to recover solvent to obtain extract, separating Gel (MCI Gel) by medium pressure chromatography, and eluting with methanol-water gradient (2:8, 5:5, 8:2, 10:0, v/v), wherein each gradient has 4-6 column volumes to obtain 3 components (Fr.1-Fr., 4). And carrying out gradient elution (100:1- & gt 1:1, v/v) on the collected Fr.2 part by adopting normal phase silica gel column chromatography (the mass ratio of the silica gel to the sample is 20:1) by taking chloroform-methanol as an eluent to obtain five parts to obtain 5 components Fr.2.1-Fr.2.5. The method comprises the steps of carrying out a first treatment on the surface of the Fr.2.4 gel column chromatography (Sephadex LH-20) with chloroform: methanol (1:1, v/v) is used as an eluent, and then the sarcoglabra A, the sarcoglabra B and the sarcoglabra C are prepared by a semi-preparative High Performance Liquid Chromatography (HPLC) with acetonitrile-water as a mobile phase and a gradient elution method (the concentration of acetonitrile is 30% -55%, v/v).
Example 4: preparation method of compound sarcglabralides A-C
The whole plant (dried) of Hainanensis (S.hainanensis), after crushing, extracting with methanol at room temperature according to the volume ratio of organic solvent to plant material of 4:1 for 2 times by ultrasonic wave, merging the extracting solution, and then distilling under reduced pressure to obtain pasty extract. Subjecting the pasty extract to silica gel column chromatography, and gradient eluting with petroleum ether-ethyl acetate (1:0→0:1, v/v) to obtain 8 components (Fr.1-Fr.8). The collected fraction Fr.5 was subjected to medium pressure chromatography separation Gel (MCI Gel) column chromatography, and eluted with a methanol-water gradient (2:8.fwdarw.10:0, v/v) to give 5 fractions Fr.5.1-Fr.5.5. Fr.5.2 is subjected to gel column chromatography (Sephadex LH-20) with methanol as an eluent, then semi-preparative High Performance Liquid Chromatography (HPLC) is performed with acetonitrile-water as a mobile phase, and a gradient elution method (30% -45% acetonitrile concentration, v/v) is adopted to prepare the sarcoglabra A-C.
Example 5: anti-inflammatory Activity of Sarcabralides A-C
Nitric Oxide (NO) has a wide and important biological regulatory function and plays an important role in inflammation, tumor, cardiovascular system and the like. When immune cells are stimulated by microbial endotoxins, inflammatory mediators, etc., a large amount of inducible nitric oxide synthase (induced NO synthase, iNOS) is produced, which produces NO to mount an immune response, and thus inhibition of NO production is a direct indicator of the anti-inflammatory activity of the compounds.
RAW264.7 cells were inoculated into 96-well plates, induced stimulation with 1. Mu.g/ml LPS, and treatment with test compound (final concentration 50. Mu.g/ml) was performed, and a drug-free group and an L-NMMA positive drug group were set as controls. After overnight incubation of the cells, the medium was taken to detect NO production and absorbance was measured at 570 nm. Cell viability assays were performed by adding MTS to the remaining medium, excluding the toxic effects of the compounds on the cells.
NO generation inhibition (%) = (non-drug treated group OD 570nm Sample set OD 570nm ) Non-drug treatment group OD 570nm ×100％
IC 50 (semi-inhibitory concentration) per Reed&The Muench method.
The experimental results are shown in tables 1 and 2, and the sarcoglabra-C has remarkable NO generation resisting activity and IC thereof 50 The values were 16.60, 13.43 and 17.19. Mu.M, respectively, 2.4-3.1 times that of the positive control L-NMMA (41.33. Mu.M).
TABLE 1 inhibition of NO production by Sarcabaraalides A-C (%)
a L-NMMA as positive control
TABLE 2 IC for inhibiting NO production by Sarcabaraalides A-C 50 Value of
a L-NMMA was used as a positive control.

Claims (7)

1. Linderane dimeric sesquiterpenes are characterized by the following structural formula I or II:
2. a pharmaceutical composition for the treatment of inflammatory diseases, characterized by comprising, as an active pharmaceutical ingredient, compound sarcglabralides A or B of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, adjuvant or excipient.
3. Use of a compound of linderane dimeric sesquiterpene or a pharmaceutically acceptable salt thereof according to claim 1 in the manufacture of a medicament for the treatment of inflammatory diseases.
4. A process for preparing the linderane dimeric sesquiterpenes according to claim 1, comprising the steps of:
1) Drying and pulverizing herba Pileae Scriptae (Sarcandra), extracting with organic solvent, and desolventizing to obtain herba Pileae Scriptae plant material extract;
2) Sequentially subjecting the herba Pileae Scriptae plant material extract to column chromatography and semi-preparative high performance liquid chromatography to obtain compounds sarcglabralides A and B,
wherein the sarcandra plant material is root, stem, branch, leaf, fruit or whole plant of sarcandra glabra (S.glabra) or sarcandra glabra (S.hainanensis),
the organic solvent comprises ethyl acetate, methanol or 95% ethanol, and
the column chromatography comprises normal phase silica gel column chromatography, reversed phase silica gel column chromatography, resin column chromatography, gel column chromatography, medium pressure chromatography separation gel and preparation or semi-preparation high performance liquid chromatography.
5. The method according to claim 4, wherein in step 1), the organic solvent extraction method comprises an organic solvent room temperature cold leaching method, an organic solvent heating reflux method or an ultrasonic method; the volume ratio of the organic solvent to the sarcandra plant material is 1:1 to 5:1.
6. A method as claimed in claim 5, wherein the volume ratio of organic solvent to sarcandra plant material is 3:1.
7. The method according to claim 4, wherein in the step 2), the eluent used for the column chromatography comprises one or a combination of two or more of petroleum ether, chloroform, methylene chloride, ethyl acetate, acetone, ethanol, methanol, n-butanol, acetonitrile, water and formic acid; the normal phase silica gel column chromatography adopts petroleum ether and ethyl acetate to carry out gradient elution according to the volume ratio of 1:0 to 0:1; or normal phase silica gel column chromatography adopts chloroform and methanol to carry out gradient elution according to the volume ratio of 100:1 to 1:1; reversed phase silica gel column chromatography is reversed phase RP-18 column chromatography, acetonitrile and water are adopted for gradient elution according to the volume ratio of 2:8 to 1:0; eluting by adopting methanol or chloroform and methanol, wherein the volume ratio of chloroform to methanol is 1:1; the medium-pressure chromatographic separation gel is eluted with methanol and water in the volume ratio of 2:8 to 1:0; the preparative or semi-preparative high performance liquid chromatography uses acetonitrile and water to perform gradient elution according to the volume ratio from 2:8 to 6:4.
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