CN114057764B - Hignathane-type dimerized sesquiterpene with anti-inflammatory activity and its preparation method and use - Google Patents

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

Hignathane-type dimerized sesquiterpene with anti-inflammatory activity and its preparation method and use

Technical field

The present invention belongs to the technical field of phytochemistry and medicine. Specifically, it relates to three anti-inflammatory dimeric sesquiterpenes sarcglabralides A-C with anti-inflammatory activity, containing any one or a combination of two or three of them. The composition also relates to a preparation method of the compounds sarcglabralides A-C and their application in preparing anti-inflammatory drugs.

Background technique

Inflammation is an active defense response of living tissues with vascular systems to endogenous or exogenous stimuli. Under normal physiological conditions, moderate and controllable inflammation can help the body clear and repair damage. Under pathological conditions, uncontrollable and excessive inflammation can lead to tissue and organ damage. Long-term persistent chronic inflammation has many hazards to human health, such as: inducing the occurrence of cancer - Helicobacter pylori infectious inflammation, EB virus infectious inflammation, and hepatitis virus inflammation are respectively related to gastric cancer, nasopharyngeal cancer, and liver cancer; endangering cardiovascular health —Chronic inflammation caused by fatty plaques in blood vessels can form blood clots and eventually lead to pulmonary arterial hypertension, chronic obstructive pulmonary disease, emphysema, and heart disease; induce Alzheimer’s disease—As we age, the inflammatory response leads to immune Disturbances in the system's regulatory mechanism increase the occurrence of early Alzheimer's disease; in addition, long-term excessive inflammatory response can also cause diabetes and its complications, Parkinson's disease, macular degeneration, osteoarthritis, etc. With the continuous acceleration of urbanization and aging and changes in lifestyles, inflammatory diseases have become one of the important threats to the health of the global population. They are spreading around the world and have evolved into a serious hazard to human health and sustainable social and economic development. important public health issues.

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, inflammatory factors (IL-1, IL-6, TNF-α, amines, 5-hydroxytryptamine), etc. These physiological and pathological changes form an inflammatory response network. NO (nitric oxide) plays a key regulatory role in inflammatory responses, especially in the occurrence and signaling of inflammatory responses. After NO is catalyzed by nitric oxide synthase (NOS) to generate L-arginine, it acts on self-generated cells or diffuses to neighboring cells and binds to its target receptors (transcription factors, protein kinases, etc.), thereby exerting a series of regulation effect. In the process of immune inflammation, NO in the body can activate NF-κB at high concentrations and induce the production of pro-inflammatory cytokines such as TNF-α and IL-1β. The production of IL-1 and TNF-α can activate iNOS. (Inducible nitric oxide synthase), promotes the body to produce more NO, sustains the secretion of cytokines and its spontaneous expression of NO, and makes the inflammatory response longer-lasting and more intense. Therefore, inhibiting the production of NO during the inflammatory reaction can improve the inflammatory symptoms.

At present, the treatments for inflammation are mainly steroids (glucocorticoids: such as dexamethasone, prednisone, hydrocortisone, etc.) and non-steroidal drugs (NSAIDS: such as aspirin, ibuprofen, indomethacin etc.) chemically synthesized drugs, which all have a variety of side effects, such as gastrointestinal reactions, cardiotoxicity, hepatotoxicity, nephrotoxicity and immune dysfunction. Therefore, it is necessary and urgent to continue to search for new anti-inflammatory lead compounds and to research and develop anti-inflammatory drugs with good efficacy and few side effects.

Natural products have gone through a long and lengthy evolutionary process in nature. The novelty of their chemical structures and the diversity of biological activities are one of the important resources for drug research and development. There is huge potential in finding lead compounds with anti-inflammatory activity from natural products. .

Contents of the invention:

In view of this, the object of the present invention is to use three new igiganane-type dimeric sesquiterpene natural products, namely the compounds sarcglabralide A, sarcglabralide B and sarcglabralide C, and these three compounds as active ingredients in the preparation of anti-inflammatory drugs. It provides natural products with good efficacy and few side effects for the treatment of inflammatory diseases.

The object of the present invention can be achieved through the following technical solutions.

In a first aspect, the present invention provides higenazane-type dimerized sesquiterpenes, namely compounds sarcglabralides A, B and C, which have the structural formulas shown in the following formulas I, II and III respectively:

In a second aspect, the present invention provides a pharmaceutical composition for the treatment of inflammatory diseases, which contains as an active pharmaceutical ingredient the higenazane-type dimeric sesquiterpene compound sarcglabralides A, B or C or a combination thereof, or a pharmaceutical composition thereof. acceptable derivatives, as well as pharmaceutically acceptable carriers, adjuvants and excipients.

In a third aspect, the present invention relates to the use of higenazane-type dimeric sesquiterpene compounds sarcglabralides A, B or C or pharmaceutically acceptable derivatives thereof in the preparation of medicaments for the treatment of inflammatory diseases.

In a fourth aspect, the present invention provides a method for preparing hilagane-type dimeric sesquiterpene compounds sarcglabralidesA, B or C, comprising the following steps:

1) Dry and pulverize the Sarcandra plant material, extract it with an organic solvent and then desolubilize it to obtain the Sarcandra plant material extract;

2) Use column chromatography, medium-pressure separation gel and semi-preparative high-pressure liquid chromatography in sequence to obtain compounds sarcglabralides A-C from the plant material extract of the genus Sarcglabra.

In a preferred embodiment, the organic solvent includes at least one of petroleum ether, chloroform, dichloromethane, ethyl acetate, acetone, ethanol, methanol, n-butanol, acetonitrile and formic acid.

Compared with the prior art, the present invention has obvious beneficial effects. Specifically, the higenazane-type dimeric sesquiterpenes sarcglabralides A–C of the present invention have significant anti-inflammatory activity and can be used to treat inflammatory diseases.

Description of the drawings

Figure 1 is the chemical structural formula of sarcglabralides A-C, the higenane-type dimeric sesquiterpene compound of the present invention.

Figure 2 is a diagram showing the ECD (electron circular dichroism) calculation results of the absolute configuration determined for the compound of the present invention, the higenoid-type dimeric sesquiterpene sarcglabralides A–C.

Detailed ways

In order to facilitate understanding of the invention, the invention will be described more fully below, and preferred embodiments of the invention will be given. However, the invention may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that a thorough understanding of the present disclosure will be provided.

Unless otherwise defined, 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 is for the purpose of describing specific embodiments only and is not intended to limit the scope of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

According to a first aspect, the present invention provides higenazane-type dimeric sesquiterpene compounds sarcglabralides A, B and C, whose structural formulas are shown in the following formulas I, II and III respectively:

According to a second aspect of the present invention, the present invention provides a pharmaceutical composition for treating inflammatory diseases, which contains as an active pharmaceutical ingredient an iginadine-type dimeric sesquiterpene compound sarcglabralides A, B or C or a pharmaceutical composition thereof. Acceptable derivatives, and pharmaceutically acceptable carriers, adjuvants or excipients.

In embodiments of the pharmaceutical composition of the present invention, inflammatory diseases include, but are not limited to: skin inflammation, rheumatic diseases, cardiovascular diseases, autoimmune diseases, etc.

In the embodiment of the pharmaceutical composition of the present invention, the higenazane-type dimeric sesquiterpene as the active ingredient can be any one, any two or three of the compounds sarcglabralides A to C.

Those skilled in the art will understand 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, can also be used in the pharmaceutical compositions of the present invention.

The pharmaceutically acceptable salt may be, for example, a pharmaceutically acceptable salt with a base.

Pharmaceutically acceptable base 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 amines, triethylamine, tripropylamine, and ethanolamine and triethanolamine.

Therefore, in the embodiment of the pharmaceutical composition of the present invention, the active pharmaceutical ingredient can be any one compound, a mixture of any two or three of sarcglabralides A-C, or a base addition salt, solvate, hydrate, etc. thereof derivative.

Suitable pharmaceutical excipients are well known to those skilled in the art. Pharmaceutical carriers or excipients are one or more solid, semi-solid and liquid diluents, fillers and pharmaceutical product auxiliaries, including but not limited to fillers (diluents), lubricants (glidants or anti-adhesion agents) ), dispersants, wetting agents, adhesives, solubilizers, antioxidants, bacteriostatic agents, emulsifiers, disintegrants, etc. Binders include syrup, gum arabic, gelatin, sorbitol, tragacanth, cellulose and its derivatives (such as microcrystalline cellulose, sodium carboxymethylcellulose, ethylcellulose or hydroxypropylmethylcellulose, etc.) , gelatin slurry, syrup, starch slurry or polyvinylpyrrolidone, etc.; fillers include lactose, sugar powder, dextrin, starch and its derivatives, cellulose and its derivatives, inorganic calcium salts (such as calcium sulfate, calcium phosphate, phosphoric acid Hydrocalcium, precipitated calcium carbonate, etc.), sorbitol or glycine, etc.; lubricants include micronized silica gel, magnesium stearate, talc, aluminum hydroxide, boric acid, hydrogenated vegetable oil, polyethylene glycol, etc.; disintegrants include starch and Its derivatives (such as sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch, etc.), polyvinylpyrrolidone or microcrystalline cellulose, etc.; wetting agents include dodecane base sodium sulfate, water or alcohol, etc.; antioxidants include sodium sulfite, sodium bisulfite, sodium metabisulfite, dibutylbenzoic acid, etc.; bacteriostatic agents include 0.5% phenol, 0.3% cresol, 0.5% chlorobutanol etc.; emulsifiers include polysorbate-80, sorbitan, lecithin, soybean lecithin, etc.; solubilizers include Tween-80, bile, glycerin, etc.

When the higenazane-type dimerized sesquiterpene of the present invention is used as a medicine, it can be administered directly or in the form of a pharmaceutical composition. In the pharmaceutical composition of the present invention, the pharmaceutical composition may contain 0.1-99% of sarcglabralides A, B or C, preferably 0.5-90%, based on the total weight of the pharmaceutical composition.

The "effective amount" of the higenazane-type dimeric sesquiterpene compound Sarcglabralides A, B or C or its pharmaceutically acceptable derivatives such as base addition salts refers to the amount sufficient to achieve the desired biological effect. It is understood that effective dosages will depend on the age, sex, health and weight of the recipient. Generally, the effective amount is determined by the person administering the treatment, such as the treating physician.

The pharmaceutical composition of the present invention can be administered in a dosage per unit body weight. All pharmaceutical compositions containing igiganane-type dimeric sesquiterpenes sarcglabralides A–C or their pharmaceutically acceptable derivatives such as base addition salts as active ingredients are prepared into various dosage forms using methods recognized in the pharmaceutical and food fields, For example, liquid preparations (injections, suspensions, emulsions, solutions, syrups, etc.), solid preparations (tablets, capsules, granules, granules, etc.), sprays, aerosols, etc. The pharmaceutical composition of the present invention can be used to treat inflammatory diseases via 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 the higenazane-type dimeric sesquiterpene compounds sarcglabralides A–C or pharmaceutically acceptable derivatives thereof of the present invention in the preparation of medicaments for the treatment of inflammatory diseases.

In embodiments of the present 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 method for preparing hilagane-type dimeric sesquiterpene compounds sarcglabralidesA–C, which includes the following steps:

1) Dry and crush Sarcandra plant material, extract it with an organic solvent and then remove it to obtain Sarcandra plant extract;

2) Use column chromatography, medium-pressure chromatography to separate gel and semi-preparative high-performance liquid chromatography in sequence for the plant extract of the genus Sarcglabra to obtain sarcglabralides A–C.

In specific embodiments, the plant material of the genus Coral can be various parts of the plant of the genus Coral, such as roots, stems, branches, leaves or fruits, or the whole plant. In embodiments of the present invention, the plant material of the genus S. glabra can be various parts (eg, roots, stems, branches, leaves or fruits) or the whole plant of S. glabra, S. hainanensis. In a preferred embodiment of the invention, the glabra plant material is derived from S. glabra.

In an embodiment of the present invention, the organic solvent used for extraction in step 1) includes at least one of petroleum ether, chloroform, dichloromethane, ethyl acetate, acetone, ethanol, methanol, n-butanol, acetonitrile, and formic acid, preferably , the organic solvent is methanol, ethyl acetate or 95% ethanol.

In embodiments of the present invention, the extraction method may be organic solvent cold immersion extraction, heated reflux extraction or ultrasonic extraction, etc.

In an embodiment of the present invention, the number of times that the plant material of the genus Coral is extracted with an organic solvent is at least 1 time, such as 2, 3, 4 times, preferably 3 times.

In embodiments of the present invention, the volume ratio of organic solvent to dry Herba plant material is 1:1 to 5:1, such as 1:1, 2:1, 3:1, 4:1, 5:1 , and the ratio between any two of the above ratios, such as 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 includes:

In specific embodiments of the present invention, in step 2), the column chromatography includes normal phase silica gel column chromatography, reversed phase silica gel column chromatography (such as RP-18), gel column chromatography (such as Sephadex LH -20), medium pressure chromatographic separation gels (e.g. MCIGel CHP20P) and preparative or semi-preparative high performance liquid chromatography (HPLC).

In a specific embodiment of the present invention, in step 2), the eluent used in column chromatography includes petroleum ether, chloroform, dichloromethane, ethyl acetate, acetone, ethanol, methanol, n-butanol, acetonitrile, water and One or a combination of any two of formic acids; normal phase silica gel column chromatography uses petroleum ether and ethyl acetate in a gradient elution from 1:0 to 0:1 in a volume ratio, such as 1:0, 50:1, 25 :1, 10:1, 5:1, 0:1; or normal phase silica gel column chromatography using chloroform and methanol in a gradient elution from 100:1 to 1:1 in a volume ratio, such as 100:1, 50:1, 30:1, 10:1, 1:1; reversed-phase RP-18 column chromatography using gradient elution of methanol and water in a volume ratio from 2:8 to 1:0; gel column chromatography, such as Sephadex LH-20 Gel column chromatography uses methanol, chloroform-methanol (v:v 1:1) to elute; medium pressure chromatography separation gel (MCI Gel) uses methanol and water in a gradient wash from 2:8 to 1:0 in a volume ratio. Desorption; Preparative or semi-preparative HPLC uses acetonitrile and water in a volume ratio of gradient elution from 2:8 to 6:4.

The preferred embodiments of the present invention will be described in detail below 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. Those skilled in the art can make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention, and all of these modifications and substitutions fall within the scope of the claims of the present invention.

The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, etc. used in the following examples can all be obtained through commercial channels unless otherwise specified.

Agilent UPLC/Q-Tof liquid mass spectrometry was measured; ultraviolet spectrum (UV) was measured in methanol with ShimazuUV-2401PC ultraviolet spectrometer; HPLC analysis and preparation used Agilent 1260 or 1100 high performance liquid chromatography, and the chromatographic column was AgilentZORBAX -SB-C18 chromatography In the following examples, ¹ H, ¹³ C NMR and 2D NMR spectra were measured on a BrukerDRX-500 nuclear magnetic resonance instrument; electrospray ionization mass spectrometry (ESI-MS) was measured by Waters XevoTQ-S triple quadrupole Determined by tandem mass spectrometer or Bruker HTC/Esquire liquid phase-ion trap chromatography mass spectrometer; high-resolution mass spectrometry (HR-ESI-MS) by column (5μm; 4.6×150mm) or Agilent ZORBAX-SB-C18 chromatography Column (5μm; 9.4×250mm); normal-phase silica gel (200–300 mesh) used for column chromatography and thin-layer chromatography plates are products of Qingdao Ocean Chemical Factory; thin-layer chromatography was observed through 10% FeCl ₃ -ethanol solution. Spots; Sephadex LH-20 is a product of GE Healthcare; reversed phase materials Rp-8 and Rp-18 are products of Merck. MCI Gel CHP20P is a product of Mitsubishi Chemical of Japan.

Example 1: Preparation method 1 of compounds sarcglabralides A–C

Grass coral (S.glabra) roots (dry), crushed and extracted with ethyl acetate at room temperature at a volume ratio of organic solvent to plant material of 3:1, cold-soaked 4 times, 2 days/time, and the extracts were combined Distill under reduced pressure to obtain a paste extract. The paste extract was subjected to silica gel column chromatography and eluted with petroleum ether-ethyl acetate gradient (40:1→1:1, v/v) to obtain 6 components (Fr.1–Fr.6). The collected Fr.4 fraction was subjected to RP-18 reversed-phase column chromatography and eluted with methanol-water gradient (2:8→1:0, v/v) to obtain 6 components Fr.4.1–Fr. 4.6. Fr.4.3 uses gel column chromatography (Sephadex LH-20) with chloroform:methanol (1:1, v/v) as the eluent, and then passes through semi-preparative high performance liquid chromatography (HPLC) with acetonitrile-water As the mobile phase, use the isocratic elution method (acetonitrile concentration 45%, v/v) to prepare sarcglabralideA and sarcglabralideB. Fr.4.5 adopts normal phase silica gel column chromatography, and uses chloroform-methanol (30:1, v/v) to elute to obtain sarcglabralideC.

Physical constants and spectral data of SarcglabralideA: yellow gel; 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 of Sarcglabralide B: yellow gel; 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/z715.2723[M+Na] ⁺ (calcd. for C ₃₈ H ₄₄ O ₁₂ Na, 715.2725).

Physical constants and spectral data of Sarcglabralide C: yellow gel; 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/z659.2465[M+Na] ⁺ (calcd. for C ₃₅ H ₄₀ O ₁₁ Na,659.2463).

Example 2: Preparation method two of compounds sarcglabralidesA–C

The whole plant of S. glabra (dried) was crushed and extracted with 95% ethanol at a volume ratio of 2:1 between organic solvent and plant material, heated and refluxed three times for 2 hours each time. The extracts were combined and distilled under reduced pressure. Recover to a small volume (ethanol concentration is about 25%-30%), then pass through the LS-700B macroporous resin column, first use 5 column volumes of distilled water to elute to remove sugar, then use 75% ethanol to elute, and recover the ethanol solvent to obtain Paste extract. The paste was subjected to MCI column chromatography, using methanol-water gradient elution (3:7, 5:5, 8:2, v/v), with each gradient having 4 to 6 column volumes to obtain 3 components (Fr .1–Fr.3). The collected Fr.2 part was subjected to normal phase silica gel column chromatography (the mass ratio of silica gel dosage to sample was 20:1), and gradient elution was performed using petroleum ether-ethyl acetate as the eluent (each gradient was 3 to 5 column volumes), six parts were obtained: 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: acetic acid Ethyl ester = 5:1, v/v); Fr.2.6 (petroleum ether: ethyl acetate = 0:1, v/v); Fr.2.5 uses gel column chromatography (Sephadex LH-20), with chloroform : Methanol (1:1, v/v) is used as the eluent, and then through semi-preparative high performance liquid chromatography (HPLC), using acetonitrile-water as the mobile phase, using a gradient elution method (acetonitrile concentration 20% → 60% , v/v) to prepare sarcglabralide A, sarcglabralide B and sarcglabralide C.

Example 3: Preparation method three of compounds sarcglabralidesA–C

S. glabra branches and leaves (dried) were crushed and cold-soaked with methanol at room temperature at a volume ratio of organic solvent to plant material of 5:1, 3 times, 2 days/time, and the extracts were combined and decompressed. Distillation gives a paste-like extract. The extracts were combined and distilled under reduced pressure to recover the solvent to obtain an extract. The extract was subjected to medium pressure chromatography separation gel (MCI Gel) column chromatography, and was eluted with a methanol-water gradient (2:8, 5:5, 8:2, 10:0, v/v), 4 to 6 column volumes per gradient, and 3 components (Fr.1–Fr.,4) were obtained. The collected Fr.2 part was subjected to normal phase silica gel column chromatography (the mass ratio of silica gel dosage to sample was 20:1), and chloroform-methanol was used as the eluent for gradient elution (100:1→1:1, v/v), five parts were obtained to obtain 5 components Fr.2.1–Fr.2.5. ; Fr.2.4 uses gel column chromatography (Sephadex LH-20) with chloroform: methanol (1:1, v/v) as the eluent, and then passes through semi-preparative high performance liquid chromatography (HPLC) with acetonitrile- Water was used as the mobile phase, and the gradient elution method (acetonitrile concentration 30% → 55%, v/v) was used to prepare sarcglabralide A, sarcglabralide B and sarcglabralide C.

Example 4: Preparation method four of compounds sarcglabralides A–C

The whole plant of S. hainanensis (dried) was crushed and extracted with methanol at room temperature at a volume ratio of organic solvent to plant material of 4:1. Ultrasonic extraction was performed twice. The extracts were combined and distilled under reduced pressure to obtain a paste. Extract. The paste extract was subjected to silica gel column chromatography, using petroleum ether-ethyl acetate gradient elution (1:0 → 0:1, v/v) to obtain 8 components (Fr.1–Fr.8). The collected Fr.5 fraction was subjected to medium pressure chromatography separation gel (MCI Gel) column chromatography, and was eluted with a methanol-water gradient (2:8→10:0, v/v) to obtain 5 components Fr. .5.1–Fr.5.5. Fr.5.2 was subjected to gel column chromatography (SephadexLH-20), using methanol as the eluent, and then through semi-preparative high performance liquid chromatography (HPLC), using acetonitrile-water as the mobile phase, and using a gradient elution method ( Sarcglabralide A–C can be prepared by acetonitrile concentration 30% → 45%, v/v).

Example 5: Anti-inflammatory activity of Sarcglabralides A–C

Nitric oxide (NO) has extensive and important biological regulatory functions, and plays an important role in inflammation, tumors, and the cardiovascular system. When immune cells are stimulated by microbial endotoxins, inflammatory mediators, etc., they will produce a large amount of induced NO synthase (iNOS) to produce NO for immune response. Therefore, inhibiting NO production is a direct result of the anti-inflammatory activity of compounds. index.

RAW264.7 cells were seeded into a 96-well plate, induced and stimulated with 1 μg/ml LPS, and the compound to be tested (final concentration 50 μg/ml) was added for treatment. A drug-free group and an L-NMMA positive drug group were set up as controls. After the cells were cultured overnight, the culture medium was taken to detect NO production, and the absorbance value was measured at 570 nm. MTS was added to the remaining culture medium to detect cell viability to eliminate the toxic effects of compounds on cells.

NO production inhibition rate (%) = (non-drug treated group OD _570nm – sample group OD _570nm )/non-drug treated group OD _570nm × 100%

IC ₅₀ (half inhibitory concentration) is calculated according to Reed & Muench method.

The experimental results are shown in Table 1 and Table 2. sarcglabralidesA–C have significant anti-NO production activity. Their IC ₅₀ values are 16.60, 13.43 and 17.19 μM respectively, which are 2.4– 2.4% higher than the positive control drug L-NMMA (41.33 μM). 3.1 times.

Table 1. Inhibition rate of NO production by sarcglabralidesA–C (%)

^a L-NMMA is the positive control drug

Table 2. IC ₅₀ values of sarcglabralidesA–C for inhibiting NO production

^a L-NMMA is the positive control drug.

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.