Acylphloroglucinol heteroterpenoid compounds and their applications

Abstract

The invention belongs to the field of plant chemistry and pharmaceutical technology, and relates to an acylphloroglucinol terpenoid compound, a composition comprising the compound, the use of the compound in preparing a drug for resisting Gram-positive bacteria, and a preparation method of the compound. The acylphloroglucinol terpenoid of the invention has obvious inhibitory activity against Gram-positive bacteria, especially Methicillin-resistant Staphylococcus aureus (MRSA), Propionibacterium acnes and Staphylococcus epidermidis.

Description

Technical Field

[0001] The present invention belongs to the technical field of plant chemistry and pharmacology, and in particular, to an acylphloroglucinol heteroterpenoid compound (Formula 1-9) separated from Eucalyptus plants, a composition containing the compound, and application of the compound in preparing an anti-Gram-positive bacteria drug. Background Art

[0002] Antibiotics are the cornerstone of the modern healthcare system. Effective infection control has promoted the progress of modern medicine: invasive surgery has become routine; chemotherapy that destroys the immune system has been introduced into the "war" against cancer; organ transplants have prolonged life; and the complete replacement of deteriorating joints, diseased corneas, and burned skin has improved the quality of life. However, the rapid emergence and widespread spread of antibiotic resistance (AMR) has made diseases that were once easily curable deadly again. For example, methicillin-resistant Staphylococcus aureus (MRSA) causes chronic and recurrent infections, leading to severe illness and high mortality in humans and animals. The emergence of multi-drug resistant (MDR) bacteria has made almost all antibiotics ineffective, and we may face the dilemma of having no drugs available.

[0003] Unfortunately, the rise in global drug resistance is accompanied by the failure of research and development of new antimicrobial drugs. Since the discovery of daptomycin and linezolid in the 1980s, no new backbone-type antibiotics have been discovered. Therefore, innovative methods are urgently needed to improve the efficiency of the discovery of new antibiotics to address the drug resistance crisis. Historically, since Alexander Fleming discovered penicillin from Penicillium (fungus) in the 1920s, the US FDA has approved 162 antibiotics between 1981 and 2019, of which about 50% are from or derived from natural products, while synthetic combinatorial compound libraries have failed to meet expectations in the discovery of new antibiotics, suggesting that the biologically relevant chemical space with extensive chemical diversity and complexity catalyzed by proteins has the potential to discover new antibiotics because it has interacted with complex life phenomena during evolution.

[0004] Eucalyptus is a plant of the Myrtaceae family, native to Australia, with more than 800 species. Nearly 80 species were introduced into my country in 1890 and are widely distributed in Yunnan, Guangxi, Guangdong and other places. Eucalyptus is one of the three major timber afforestation tree species in the world. The plantation area of ​​artificial forests in my country exceeds 4.4 million hectares, and the resource volume is huge. The leaves, fruits and resins of E. robusta, E. globulus, E. maideni, E. citriodora, E. tereticornis and E. exserta are used by the people to treat influenza, eczema, dysentery and the like, and have a good traditional medicinal background. In addition to essential oils, plants of this genus are also rich in non-volatile characteristic components - phloroglucinol derivatives. However, there are few reports on the antibacterial activity of such compounds isolated from Eucalyptus plants. Summary of the invention

[0005] In view of this, the object of the present invention is to provide an acylphloroglucinol heteroterpenoid compound shown in Formula 1-9 or a pharmaceutically acceptable derivative, a prodrug and a preparation method thereof, a composition comprising the compound or a pharmaceutically acceptable derivative or prodrug thereof, and the use of such a compound or a pharmaceutically acceptable derivative or prodrug thereof in the preparation of a drug against Gram-positive bacteria.

[0006] In a first aspect, the present invention provides an acylphloroglucinol heteroterpenoid compound or a pharmaceutically acceptable derivative or prodrug thereof, wherein the structural formula of the compound is shown in Formula 1-9 below.

[0007]

[0008] In a second aspect, the present invention provides a pharmaceutical composition comprising as an active pharmaceutical ingredient one or more of the acylphloroglucinol terpenoid compounds of formula 1-9 or their pharmaceutically acceptable salts, prodrugs, and a pharmaceutically acceptable carrier, adjuvant or excipient.

[0009] In a third aspect, the present invention provides the use of acylphloroglucinol heteroterpenoid compounds represented by formula 1-9, and pharmaceutically acceptable derivatives or prodrugs thereof in the preparation of drugs against Gram-positive bacteria.

[0010] In a fourth aspect, the present invention provides a method for preparing the acylphloroglucinol terpenes described above, comprising the following steps:

[0011] 1) drying and crushing the eucalyptus plant material, extracting with an organic solvent and then desolventizing to obtain a eucalyptus plant extract; and

[0012] 2) The eucalyptus plant extract is sequentially subjected to column chromatography and thin layer chromatography to obtain the acylphloroglucinol terpenoid (Formula 1-9).

[0013] Compared with the prior art, the present invention has obvious beneficial effects. Specifically, the acylphloroglucinol terpenes provided by the present invention have significant inhibitory activity against methicillin-resistant Staphylococcus aureus (MRSA), Propionibacterium acnes (C.acnes) and Staphylococcus epidermidis (S.epidermidis), wherein the minimum inhibitory concentration (MIC) of the compound eucalrobusone F (1) against methicillin-resistant Staphylococcus aureus (MRSA440) is better than that of the positive control drug vancomycin; the minimum inhibitory concentration (MIC) of the compounds eucalypglobulusal H (2), eucalrobusone F (1) and eucarobustol D (7) against Propionibacterium acnes is better than that of the positive control drug tetracycline; the compounds eucalypglobulusal H (2), eucalrobusone F (1), eucarobustol F (3), eucarobustol H (4), macrocarpal O (5), eucalteretial E (6), macrocarpal H (8) and macrocarpal The minimum inhibitory concentration (MIC) of Q(9) against Staphylococcus epidermidis was better than that of the positive control drug tetracycline. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Schematic diagram of structural formula 1-9 of the acylphloroglucinol heteroterpenoid compound of the present invention. DETAILED DESCRIPTION

[0015] In order to facilitate the understanding of the present invention, the present invention will be described more fully below, and preferred embodiments of the present invention are provided. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0016] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present invention belongs. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the scope of the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0017] According to the first aspect of the present invention, the present invention provides an acylphloroglucinol heteroterpenoid compound (Formula 1-9) or a pharmaceutically acceptable salt or prodrug thereof. Specifically, the acylphloroglucinol heteroterpenoid compound includes: acylphloroglucinol coupled monoterpene eucalrobusone F (1) and acylphloroglucinol coupled sesquiterpene eucalypglobulusal H (2), eucarobustol F (3), eucarobustol H (4), macrocarpal O (5), eucalteretial E (6), eucarobustol D (7), macrocarpal H (8), macrocarpal Q (9), and their structural formulas are shown in Formulas 1-9 above, respectively.

[0018] According to the second aspect of the present invention, the present invention provides a pharmaceutical composition comprising as an active pharmaceutical ingredient any one or more of the above-mentioned acylphloroglucinol terpene compounds (Formula 1-9) or pharmaceutically acceptable salts, prodrugs thereof, and pharmaceutically acceptable carriers, adjuvants or excipients.

[0019] Those skilled in the art should understand that pharmaceutically acceptable derivatives of the acylphloroglucinol terpenoid compounds (Formula 1-9) of the present invention, such as salts, esters, solvates or hydrates of the acylphloroglucinol terpenoid compounds (Formula 1-9) can also be used in the pharmaceutical composition of the present invention.

[0020] Suitable pharmaceutical excipients are well known to those skilled in the art. Pharmaceutical carriers or excipients are one or more solid, semisolid and liquid diluents, fillers and adjuvants for pharmaceutical products, including but not limited to fillers (diluents), lubricants (glidants or anti-adhesives), dispersants, wetting agents, adhesives, solubilizers, antioxidants, antibacterial agents, emulsifiers, disintegrants, etc. Binders include syrup, gum arabic, gelatin, sorbitol, tragacanth, cellulose and its derivatives (such as microcrystalline cellulose, sodium carboxymethyl cellulose, ethyl cellulose or hydroxypropyl methylcellulose, etc.), gelatin slurry, syrup, starch slurry or polyvinyl pyrrolidone, etc.; fillers include lactose, powdered sugar, dextrin, starch and its derivatives, cellulose and its derivatives, inorganic calcium salts (such as calcium sulfate, calcium phosphate, calcium hydrogen phosphate, precipitated calcium carbonate, etc.), sorbitol or glycine, etc.; lubricants include micropowdered 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.), polyvinyl pyrrolidone or microcrystalline cellulose, etc.; wetting agents include sodium lauryl sulfate, water or alcohol, etc.; antioxidants include sodium sulfite, sodium bisulfite, sodium metabisulfite, dibutyl benzoic acid, etc.; antibacterial agents include 0.5% phenol, 0.3% cresol, 0.5% trichlorobutanol, etc.; emulsifiers include polysorbate-80, sorbitan monohydrate, lecithin, soybean lecithin, etc.; solubilizers include Tween-80, bile, glycerol, etc.

[0021] When the acylphloroglucinol heteroterpenoid compound (Formula 1-9) or its pharmaceutically acceptable prodrug or derivative is used as a drug, it can be directly administered or administered in the form of a pharmaceutical composition. In the pharmaceutical composition of the present invention, the pharmaceutical composition may contain 0.1-99%, preferably 0.5-90% of the acylphloroglucinol heteroterpenoid compound (Formula 1-9) or its derivative, based on the total weight of the pharmaceutical composition.

[0022] The therapeutically effective amount of acylphloroglucinol terpenoid compound (Formula 1-9) or a pharmaceutically acceptable prodrug or derivative thereof refers to the amount of acylphloroglucinol terpenoid compound (Formula 1-9) sufficient to achieve the desired biological effect, such as inducing platelet aggregation, thereby achieving a hemostatic effect without causing significant negative or adverse side effects to the recipient. It should be understood that the effective dose will depend on the age, sex, health status and weight of the recipient. Typically, the effective amount is determined by the person administering the treatment, such as a treating physician.

[0023] The pharmaceutical composition of the present invention can be administered in the form of a dosage per unit body weight. All pharmaceutical compositions with acylphloroglucinol heteroterpenoid compounds (Formula 1-9) or pharmaceutically acceptable prodrugs and derivatives thereof as active ingredients can be prepared into various dosage forms by methods recognized in the pharmaceutical and food fields, such as 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 administered by injection (intravenous injection, intravenous drip, intramuscular injection, intraperitoneal injection, subcutaneous injection), oral administration, sublingual administration, mucosal dialysis, etc., to treat hemorrhagic diseases.

[0024] According to the third aspect of the present invention, the present invention provides the use of acylphloroglucinol heteroterpenoid compounds represented by formula 1-9 or pharmaceutically acceptable derivatives and prodrugs thereof in the preparation of drugs against Gram-positive bacteria.

[0025] In an embodiment of the present invention, Gram-positive bacteria refer to bacteria whose bodies are purple or blue-purple after Gram staining, including cocci, non-spore rods and spore rods, for example, Gram-positive bacteria from the following genera: Actinomyces, Bacillus, Clostridium, Corynebacterium, Lactobacillus, Listeria, Mycobacterium, Myxococcus, Nocardia, Staphylococcus, Streptococcus, Streptomyces and Cutibacterium.

[0026] In a preferred embodiment of the third aspect of the present invention, the Gram-positive bacteria are from the genus Staphylococcus or Propionibacterium. In a further preferred embodiment, the Staphylococcus is methicillin-resistant Staphylococcus aureus (MRSA) or Staphylococcus epidermidis. In a further preferred embodiment, the Propionibacterium is Propionibacterium acnes.

[0027] In the embodiments of the present invention, the antibacterial activity can be expressed by minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC).

[0028] According to a fourth aspect of the present invention, the present invention provides a method for preparing the above-mentioned acylphloroglucinol heteroterpenoid compound (Formula 1-9), comprising the following steps:

[0029] 1) drying and crushing the eucalyptus plant material, extracting with an organic solvent and then desolventizing to obtain a eucalyptus plant extract; and

[0030] 2) The eucalyptus plant extract is sequentially subjected to column chromatography and thin layer chromatography to obtain the acylphloroglucinol terpenoid (Formula 1-9).

[0031] In the present invention, the genus Eucalyptus refers to species in the genus Eucalyptus L.Herit of the Myrtaceae family. In a specific embodiment, the genus Eucalyptus can be selected from, for example, E. alba, E. amplifolia, E. bicolor, E. blakelyi, E. botryoides, E. camaldulenais, E. camaldulensis, E. citriodora, E. crebra, E. exserta, E. globulus, E. grandis, E. kirtoniana, E. leptophleba. , spotted eucalyptus (E.maculata), straight-stemmed blue eucalyptus (E.maideni), honey-scented eucalyptus (E.melliodora), small-capped eucalyptus (E.microcorys), paniculata (E.paniculata), rough-skinned eucalyptus (E.pellita), broad-leaved eucalyptus (E.platyphylla), multi-flowered eucalyptus (E.polyanthemos), spotted leaf eucalyptus (E.punctata), large-leaved eucalyptus (E.robusta), wild eucalyptus (E.rudis), willow-leaved eucalyptus (E.saligna), fine-leaved eucalyptus (E.tereticornis), and hairy-leaved eucalyptus (E.torelliana) One or more.

[0032] In a preferred embodiment of the present invention, the Eucalyptus plant is one or more of Eucalyptus globulus, Eucalyptus serrata ...camaldulensis or Eucalyptus globulus.

[0033] In a specific embodiment, the eucalyptus plant material can be branches and leaves, fruits or mixtures thereof of eucalyptus plants. In a preferred embodiment, the branches and leaves, fruits of eucalyptus plants are dry. In a further preferred embodiment, the eucalyptus plant material is the fruit of Eucalyptus globulus.

[0034] In step 2), column chromatography includes normal phase silica gel column chromatography, reverse phase silica gel column chromatography, gel column chromatography, MCI GelCHP20P column chromatography, preparative or semi-preparative high performance liquid chromatography (HPLC); preferably, column chromatography includes normal phase silica gel column chromatography, reverse phase RP-18 column chromatography, gel column chromatography, semi-preparative high performance liquid chromatography (HPLC).

[0035] The preferred embodiments of the present invention are described in detail below in conjunction with the accompanying drawings and examples. It should be understood that the following examples are provided only for the purpose of illustration and are not intended to limit the scope of protection of the present invention. Those skilled in the art may 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 protection claimed in the claims of the present invention.

[0036] The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.

[0037] In the following embodiments, 1 H and 13 C NMR spectra were measured on a Bruker Avance III-500 NMR spectrometer using deuterated chloroform or methanol as solvents. Normal phase silica gel (200–300 mesh) and thin layer chromatography plates for column chromatography were products of Qingdao Ocean Chemical Factory. Sephadex LH-20 was a product of GE Healthcare. Reverse phase material RP-18 and RP-18 thin layer plates were products of Merck, Germany. MCI Gel CHP20P was a product of Mitsubishi Chemical, Japan. Semi-preparative HPLC was performed on Agilent 1260 or Agilent 1100 chromatographs, using Agilent ZORBAX SB-C18 (9.4×250 mm, 5.0 μm) or Agilent ZORBAX-SB-C8 columns (9.4×250 mm, 5 μm).

[0038] Example 1: Preparation method of acylphloroglucinol terpenes (Formula 1-9)

[0039] In this embodiment, the following steps are used to prepare acylphloroglucinol terpenes (Formula 1-9):

[0040] (1) drying and crushing the straight-stemmed eucalyptus globulus fruit to obtain a plant material;

[0041] (2) using petroleum ether for cold extraction 3 times, 3 days / time, combining the extracts, and recovering the solvent under reduced pressure to obtain the straight-stemmed eucalyptus globulus extract;

[0042] (3) The straight-rod eucalyptus globulus extract was subjected to normal phase silica gel column chromatography (the mass ratio of silica gel to extract was 4:1), and gradient eluted with petroleum ether-ethyl acetate (1:0→200:1→100:1→50:1→30:1→15:1→1:1). After TLC detection and merging, 8 fractions were obtained, namely: Fr.A (petroleum ether:ethyl acetate = 1:0, v / v), Fr.B (petroleum ether:ethyl acetate = 1:0, v / v), Fr.C (petroleum ether:ethyl acetate = 200:1, v / v), Fr.D (petroleum ether:ethyl acetate = 100:1, v / v), Fr.E (petroleum ether:ethyl acetate = 50:1, v / v), Fr.F (petroleum ether:ethyl acetate = 30:1, v / v), Fr.G (petroleum ether:ethyl acetate = 15:1, v / v), and Fr.H (petroleum ether:ethyl acetate = 1:1, v / v);

[0043] (4) The Fr.C fraction was subjected to Sephadex LH-20 column (chloroform:methanol = 7:3) to remove fatty acids, and then separated using a normal phase silica gel column to obtain compounds 28 and 15;

[0044] (5) After removing fatty acids from the Fr.D fraction using Sephadex LH-20 column chromatography, the fraction was subjected to gradient elution using a reversed-phase silica gel column (Rp-18) with methanol-water (containing 1 / 1000 formic acid) as the eluent, and three fractions were collected in sections: Fr.D-1 (methanol:water = 70:30, v / v), Fr.D-2 (methanol:water = 85:15, v / v), and Fr.D-3 (methanol:water = 100:0, v / v). Among them, Fr.D-2 was separated and purified by semi-HPLC (MeCN:H 2 O=95:10, 0-30min), to obtain compound 9;

[0045] (6) Fr.F was subjected to reverse phase silica gel column chromatography (Rp-18) with methanol-water (containing 1 / 1000 formic acid) as the eluent for gradient elution, and three parts were collected in sections: Fr.F-1 (methanol: water = 70:30, v / v), Fr.F-2 (methanol: water = 80:20, v / v), Fr.F-3 (methanol: water = 90:10, v / v), and Fr.F-4 (methanol: water = 100:0, v / v). Among them, Fr.F-4 was separated and purified by semi-HPLC C18 column (MeCN: H2O = 95:10, 0-40 min) and then separated and purified by semi-HPLC-PFP column (MeCN: H2O = 95:10, 0-40 min) to obtain compound 24;

[0046] (7) Fr.H was subjected to normal phase silica gel column chromatography, eluted isocratically with petroleum ether:ethyl acetate = 20:1, and after TLC detection, 8 fractions were combined to obtain Fr.H-1 to 8. Among them, Fr.H-8 was further subjected to normal phase silica gel column chromatography, eluted isocratically with petroleum ether:ethyl acetate = 30:1, and purified by semi-HPLC (MeCN:H2O = 85:10, 0-20 min) to obtain compounds 20, 25 and 26.

[0047] (8) Fr.G was purified by Sephadex LH-20 column (chloroform:methanol=1:1) and then separated by normal phase silica gel column chromatography (petroleum ether:ethyl acetate=20:1) to obtain compounds 14, 18 and 19.

[0048] (9) Physical constants and spectral data of acylphloroglucinol terpenes (1-9):

[0049] Eucalrobusone F(1): light yellow oil; molecular formula C 23 H 28 O 5 ; 1 H NMR (CDCl 3 ,500MHz)δ H 9.96(1H,s,H-13'),14.14(1H,s,OH-1'),15.64(1H,s,OH-1'),7.05(1H,brs,H-2),7.13(1H,dd,J=7.9,1.6Hz,H-4),7. 17(1H,dd,J=7.9Hz,H-5),2.88(1H,m,H-7),1.88(1H,d,J=6.8Hz,H-8),1.88(1H,d,J=6.8Hz,H-9),3.99(2H,m,H-12'); 13 C NMR (CDCl 3 ,125MHz)δ C135.6(s,C-1),126.6(d,C-2),148.3(s,C-3),125.9(d,C-4),131.9(d,C-5),135.3(s,C-6) ,33.9(d,C-7),24.3(q,C-8),24.3(q,C-9),19.3(q,C-10),104.1(s,C-1'),168.6(s,C-2') ,104.5(s,C-3'),171.9(s,C-4'),104.0(s,C-5'),163.6(s,C-6'),206.9(s,C-7'),53.9(t ,C-8'),25.6,(d,C-9'),23.3(q,C-10'),23.3(q,C-11'),27.1(t,C-12'),192.9(d,C-13').

[0050] Eucalglobial I(2): Yellow amorphous powder; molecular formula C 28 H 38 O 5 ; 1 HNMR (500MHz, CDCl 3 )δ H 10.03(1H,s,H-8'),2.21(1H,m,H-7'β),0.92(3H,d,J=6.7Hz,Me-12'),2.36(1H,m,H-1),1.84( 1H,m,H-2α),1.67(1H,m,H-2β),1.57(1H,m,H-3α),1.38(1H,m,H-3β),1.96(1H,m,H-8α),0.92( 1H,m,H-8β),2.37(1H,m,H-9α),1.95(1H,m,H-9β),1.14(1H,t,J=10.4Hz,H-5),0.94(3H,s,Me- 12),1.04(3H,s,Me-13),4.67(1H,s,H-14a),4.61(1H,s,H-14b),0.92(3H,d,J=6.7Hz,Me-13'); 13 C NMR (125 MHz, CDCl 3 )δ C50.8(d, C-1), 26.0(t, C-2), 40.1(t, C-3), 47.0(s, C-4), 53.6(d, C-5), 27.7(d, C-6), 27.5(d, C-7), 25.5(t, C-8), 39.3(t, C-9), 154.9(s, C-10), 20.1(s, C-11), 16.7(q, C-12), 28.9(q, C-13), 105.3(t, C-14), 20.5(q, C-15), 104.5(s, C-1'), 172.8(s, C-2'), 103.3(s, C-3'), 168.0(s, C-4'), 104.0(s, C-5'), 162.2(s, C-6'), 32.8(t, C-7'), 192.1(d, C-8'), 206.6(s, C-9'), 52.9(t, C-10'), 25.0(d, C-11'), 22.7(q, C-12'), 22.7(q, C-13').

[0051] Eucarobustol F(3): Pale yellow powder; molecular formula C 28 H 40 O 6 ; 1 HNMR(500MHz, CDCl 3 ) δ H 10.20(1H, s, H-8'), 10.08(1H, m, H-7'), 13.38(1H, s, OH-4'), 13.28(1H, s, OH-6'), 3.42(1H, dd, H-9'), 2.26(1H, m, H-10α'), 1.29(1H, m, H-10β'), 0.80(3H, s, J = 6.3Hz, H-12'), 0.79(3H, s, J = 6.3Hz, H-13'), 1.82(1H, m, H-1), 1.64(1H, m, H-2α), 1.68(1H, m, H-2β), 1.34(1H, m, H-3α), 2.02(1H, m, H-3β), 1.84(1H, m, H-5), 0.53(1H, m, H-6), 0.53(1H, m, H-7), 1.58(1H, m, H-8α), 1.46(1H, m, H-8β), 1.51(1H, m, H-9α), 1.71(1H, m, H-9β), 1.06(3H, s, H-12), 1.09(3H, s, H-13), 1.19(3H, s, H-14), 0.77(3H, s, H-15); 13 C NMR(125MHz, CDCl3) δ C56.5(d, C-1), 26.1(t, C-2), 36.6(t, C-3), 49.8(s, C-4), 43.7(d, C-5), 30.3(d, C-6), 28.3(d, C-7), 20.5(t, C-8), 43.6(t, C-9), 74.2(s, C-10), 21.8(s, C-11), 30.3(q, C-12), 17.4(q, C-13), 31.4(q, C-14), 21.8(q, C-15), 107.0(s, C-1'), 171.8(s, C-2'), 106.8(s, C-3'), 171.0(s, C-4'), 110.3(s, C-5'), 171.8(s, C-6'), 192.8(d, C-7'), 192.8(d, C-8'), 37.6(d, C-9'), 36.6(t, C-10'), 28.2(d, C-11'), 25.0(q, C-12'), 21.8(q, C-13').

[0052] Eucarobustol H(4): Pale yellow powder; molecular formula C 28 H 38 O 5 ; 1 HNMR(500MHz, CDCl 3 ) δ H 10.05(1H, s, H-8'), 10.06(1H, m, H-7'), 13.36(1H, s, OH-4'), 13.29(1H, s, OH-6'), 3.32(1H, dd, J = 12.0Hz, H-9'), 2.36(1H, m, J = 12.0Hz, H-10α'), 1.20(1H, m, H-10β'), 0.83(3H, s, J = 6.2Hz, H-12'), 0.77(3H, s, J = 6.2Hz, H-13'), 5.03(1H, m, H-2), 2.07(1H, m, H-3α), 2.52(1H, m, H-3β), 2.15(1H, m, H-5), 0.44(1H, m, H-6), 0.42(1H, m, H-7), 1.82(1H, m, H-8α), 0.96(1H, m, H-8β), 1.61(1H, m, H-9α), 0.99(1H, m, H-9β), 1.06(3H, s, H-12), 1.01(3H, s, H-13), 0.88(3H, s, H-14), 1.16(3H, s, H-15); 13 C NMR(125MHz, CDCl3) δ C159.7 (d, C-1), 118.7 (t, C-2), 49.7 (t, C-3), 50.0 (s, C-4), 53.7 (d, C-5), 28.5 (d, C-6), 28.7 (d, C-7), 25.5 (t, C-8), 41.0 (t, C-9), 41.5 (d, C-10), 20.0 (s, C-11), 30.1 (q, C-12), 16.0 (q, C-13), 21.0 (q, C-14), 23.6 (q, C-15), 106.2 (s, C-1'), 166.3 (s, C-2'), 104.6 (s, C-3'), 168.0 (s, C-4'), 106.2 (s, C-5'), 170.5 (s, C-6'), 192.9 (d, C-7'), 192.2 (d, C-8'), 41.5 (d, C-9'), 36.2 (t, C-10'), 28.5 (d, C-11'), 24.3 (q, C-12'), 22.5 (q, C-13').

[0053] Macrocarpal O(5): Pale yellow powder; molecular formula C 28 H 40 O 6 ; 1 H NMR(CDCl 3 , 500 MHz) δ H 10.23 (1H, s, H-8'), 10.14 (1H, s, H-7'), 3.54 (1H, d, J = 10.5 Hz, H-9'), 1.59 (1H, m, H-10'a), 1.83 (1H, m, H-10'b), 1.62 (1H, m, H-11'), 0.80 (3H, d, J = 6.9 Hz, Me-12'), 0.82 (3H, d, J = 6.9 Hz, Me-13'), 2.89 (1H, m, H-1), 1.41 (2H, m, H-3), 1.20 (1H, m, H-2a), 1.40 (1H, m, H-2b), 2.15 (1H, m, H-7), 6.00 (1H, s, H-6), 1.26 (1H, m, H-9a), 1.99 (1H, m, H-9b), 1.64 (1H, m, H-8a), 1.74 (1H, m, H-8b), 2.01 (1H, m, H-10), 1.20 (3H, s, Me-12), 1.23 (3H, s, Me-13), 0.86 (3H, d, J = 6.8 Hz, Me-14), 1.22 (3H, s, Me-15); 13 C NMR(CDCl 3 , 125 MHz) δ C42.9(d, C-1), 29.7(t, C-2), 38.8(t, C-3), 54.5(s, C-4), 152.5(s, C-5), 124.8(d, C-6), 53.2(d, C-7), 27.5(t, C-8), 35.8(t, C-9), 37.7(d, C-10), 74.5(s, C-11), 28.0(q, C-12), 29.7(q, C-13), 18.6(q, C-14), 24.5(q, C-15), 110.4(s, C-1'), 171.5(s, C-2'), 106.3(s, C-3'), 171.0(s, C-4'), 106.0(s, C-5'), 169.8(s, C-6'), 193.0(d, C-7'), 193.1(d, C-8'), 37.7(d, C-9'), 38.0(t, C-10'), 27.9(d, C-11'), 21.9(q, C-12'), 25.0(q, C-13').

[0054] Eucalteretial E(6): Pale yellow powder; molecular formula C 28 H 40 O 6 ; 1 HNMR(CDCl 3 , 500 MHz) δ H 10.16(1H, s, H-7'), 10.17(1H, s, H-8'), 2.81(1H, q, H-9'), 1.02(1H, m, H-10'a), 2.61(1H, m, H-10'b), 1.68(1H, m, H-11'), 0.96(3H, d, Me-12'), 0.88(3H, d, Me-13'), 2.05(1H, m, H-1), 1.76(1H, m, H-2α), 1.65(1H, m, H-2β), 1.33(1H, dd, H-3α), 2.00(1H, m, H-3β), 2.00(1H, m, H-4), 2.13(1H, dd, H-5), 1.09(1H, t, H-6), 0.76(1H, td, H-7), 1.10(1H, m, H-8α), 1.54(1H, m, H-8β), 1.30(1H, m, H-9α), 2.06(1H, m, H-9β), 0.99(3H, s, Me-12), 1.50(3H, s, Me-13), 1.30(1H, m, H-14α), 2.16(1H, m, H-14β), 1.26(3H, s, H-15); 13 C NMR(CDCl 3 , 125 MHz) δC 52.6(d, C-1), 25.5(t, C-2), 36.8(t, C-3), 49.6(s, C-4), 45.1(d, C-5), 27.1(t, C-6), 48.5(d, C-7), 31.4(t, C-8), 48.6(t, C-9), 76.6(s, C-10), 152.7(s, C-11), 111.5(t, C-12), 19.9(q, C-13), 22.8(q, C-14), 21.8(q, C-15), 106.0(s, C-1'), 172.9(s, C-2'), 106.0(s, C-3'), 171.9(s, C-4'), 105.9(s, C-5'), 168.4(s, C-6'), 193.2(d, C-7'), 193.0(d, C-8'), 39.9(d, C-9'), 36.9(t, C-10'), 36.5(d, C-11'), 25.5(q, C-12'), 21.8(q, C-13').

[0055] Eucarobustol D(7): Pale yellow powder; molecular formula C 28 H 38 O 5 ; 1 H NMR(CDCl 3 , 500MHz) δ H 9.95(1H, s, H-8'), 9.96(1H, s, H-7'), 3.47(1H, m, H-9'), 1.38(1H, m, H-1), 1.56(1H, m, H-2a), 1.66(1H, m, H-2b), 1.56(1H, m, H-3a), 5.66(1H, d, H-6), 4.86(1H, brs, H-12a), 4.66(1H, brs, H-12b), 1.77(3H, s, Me-13), 1.16(3H, s, Me-14), 1.07(3H, d, Me-15), 0.80(3H, d, J = 6.1Hz, Me-12'), 0.82(3H, d, J = 6.1Hz, Me-13'); 13 C NMR(CDCl 3 , 125MHz) δ C54.9(d, C-1), 22.6(t, C-2), 35.3(t, C-3), 40.9(d, C-4), 151.5(s, C-5), 124.5(d, C-6), 43.0(d, C-7), 23.7(t, C-8), 35.3(t, C-9), 40.9(s, C-10), 149.5(s, C-11), 112.0(t, C-12), 22.6(q, C-13), 23.4(q, C-14), 23.7(q, C-15), 108.7(s, C-1'), 173.9(s, C-2'), 109.4(s, C-3'), 171.8(s, C-4'), 107.8(s, C-5'), 172.6(s, C-6'), 192.3(d, C-7'), 192.1(d, C-8'), 33.0(d, C-9'), 45.7(t, C-10'), 30.9(d, C-11'), 25.3(q, C-12'), 22.2(q, C-13').

[0056] Macrocarpal H(8): Pale yellow powder; Molecular formula C 28 H 40 O 6 ; 1 HNMR(CDCl 3 , 500 MHz) δ H 10.54(1H, s, H-8'), 10.49(1H, s, H-7'), 3.77(1H, ddd, J = 13.0, 2.4, 2.4 Hz, H-9'), 2.56(1H, d, J = 13.0 Hz, H-2a), 1.76(1H, d, J = 13.0 Hz, H-2β), 2.56(1H, ddd, J = 13.0, 2.4, 2.4 Hz, H-3a), 2.26(1H, ddd, J = 13.0, 12.5, 4.9 Hz, H-3β), 1.35(3H, s, Me-12), 1.37(3H, s, Me-13), 0.99(3H, s, Me-14), 4.86(3H, s, Me-15), 0.90(3H, d, J = 6.4 Hz, Me-12'), 1.32(3H, d, J = 6.4 Hz, Me-13'); 13 C NMR(CDCl 3 , 125 MHz) δ C54.3(d, C-1), 27.6(t, C-2), 38.5(t, C-3), 152.6(s, C-4), 52.7(d, C-5), 25.0(t, C-6), 49.9(d, C-7), 23.8(t, C-8), 38.8(t, C-9), 41.9(s, C-10), 73.4(s, C-11), 27.6(q, C-12), 28.3(q, C-13), 12.1(q, C-14), 105.3(t, C-15), 106.6(s, C-1'), 170.3(s, C-2'), 106.0(s, C-3'), 169.9(s, C-4'), 116.2(s, C-5'), 168.4(s, C-6'), 193.1(d, C-7'), 193.1(d, C-8'), 31.3(d, C-9'), 40.1(t, C-10'), 27.2(d, C-11'), 22.4(q, C-12'), 25.0(q, C-13').

[0057] Macrocarpal Q(9): Pale yellow powder; molecular formula C 28 H 40 O 6 ; 1 HNMR(CD 3 OD, 500 MHz) δ H 10.04(2H, s, H-7' / 8'), 3.35(1H, m, H-9'), 1.29(1H, m, H-10'a), 2.01(1H, m, H-10'b), 1.19(1H, m, H-11'), 0.94(3H, d, J = 5.4 Hz, Me-12'), 0.81(3H, d, J = 5.7 Hz, Me-13'), 1.58(1H, m, H-1), 1.77(1H, qd, J = 2.8, 13.5 Hz, H-2a), 2.08(1H, dd, J = 3.2, 13.4 Hz, H-2b), 1.56(1H, m, H-3a), 1.64(1H, m, H-3b), 2.43(1H, m, H-4), 5.42(1H, d, J = 2.8 Hz, H-6), 1.97(1H, m, H-7), 1.50(1H, m, H-8a), 1.59(1H, m, H-8b), 1.66(1H, m, H-9b), 1.40(1H, td, J = 3.1, 12.0, 12.8 Hz, H-9a), 1.05(3H, s, Me-12), 1.06(3H, s, Me-13), 1.17(3H, d, J = 7.5 Hz, Me-15), 1.15(3H, s, Me-14); 13C NMR (CD 3 OD,125MHz)δ C 53.2(d,C-1),21.6(t,C-2),35.1(t,C-3,41.0(d,C-4),152.3(s,C-5),122.9(d,C-6),46.6(d,C-7),21.6(t, C-8),36.7(t,C-9),40.7(s,C-10),74.3(s,C-11),27.5(q,C-12),27.8(q,C-13),23.1(q,C-14),22.4(q,C-1 5),115.9(s,C-1'),170.6(s,C-2'),106.6(s,C-3'),168.6(s,C-4'),105.9(s,C-5'),168.3(s,C-6'),193.1 (d,C-7'),193.0(d,C-8'),31.5(d,C-9'),40.2(t,C-10'),27.7(d,C-11'),22.5(q,C-12'),25.0(q,C-13').

[0058] Example 2: Antibacterial activity of acylphloroglucinol terpenes (Formula 1-9)

[0059] In order to better illustrate the pharmacological activity test results of the present invention and the superiority of the present invention, in this example, the inhibitory activity of acylphloroglucinol terpenes (Formula 1-9) against methicillin-resistant Staphylococcus aureus (MRSA), Propionibacterium acnes (C. acnes) and Staphylococcus epidermidis (S. epidermidis) was verified, but the present invention is not limited thereto.

[0060] (1) Bacterial species: Propionibacterium acnes ATCC 6919 (purchased from Guangdong Microbiological Culture Collection Center), Staphylococcus epidermidis BNCC 102555 (standard bacteria purchased from Beina Chuanglian Biotechnology Co., Ltd.), methicillin-resistant Staphylococcus aureus MRSA202 and MRSA440 (clinical drug-resistant strains isolated from the Pharmacy Department of the 920th Hospital of the Joint Logistics Support Force of the People's Liberation Army);

[0061] (2) Culture medium: Brain heart infusion medium (BHI) (purchased from Oxoid) was used to culture Propionibacterium acnes; LB medium (purchased from Guangdong Huankai Microbiological Technology Co., Ltd.) was used to culture MRSA and Staphylococcus epidermidis.

[0062] (3) Determination of minimum inhibitory concentration (MIC): The minimum inhibitory concentration was determined by the micro-culture medium multiple dilution method. 100 μL of liquid culture medium was added to a 96-well plate, 100 μL of the test compound was added to the first well, 100 μL was aspirated and injected into the next well after mixing, and 100 μL was aspirated and injected into the last well after multiple dilution. The bacteria in the logarithmic growth phase were prepared into a bacterial suspension and the concentration of the bacterial suspension was adjusted to 1×10 5 CFU / mL and 100 μL of bacterial suspension was added to a 96-well plate and incubated at 37°C for 24 h before determining the minimum inhibitory concentration.

[0063] (4) Determination of minimum bactericidal concentration (MBC): After determining the MIC, use an inoculation loop to take the bacterial solution from the first four wells before the MIC (i.e., MIC, 2MIC, 4MIC, and 8MIC) and inoculate it on an agar plate. Continue to culture for 24 hours and then determine the minimum bactericidal concentration.

[0064] Table 1. Antibacterial test results of acylphloroglucinol terpenes (1-9) Unit: μg / mL

[0065]

[0066]

[0067] As can be seen from Table 1, the acylphloroglucinol heteroterpenoid compound pair (Formula 1-9) has significant inhibitory activity against Gram-positive bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA), Propionibacterium acnes and Staphylococcus epidermidis. The antibacterial activity of some of the compounds is better than that of the positive control drug, which can provide available candidate molecules for the preparation of drugs against Gram-positive bacteria.

Claims

1. Use of acylphloroglucinol heteroterpenoid compounds and pharmaceutically acceptable salts thereof in the preparation of drugs for resisting Gram-positive bacteria, characterized in that: The structural formula of the compound is shown in Formula 1-5 and 7-9: Wherein, the Gram-positive bacteria are Staphylococcus epidermidis and Propionibacterium.

2. The use according to claim 1, characterized in that: The propionibacterium is Propionibacterium acnes (C. acnes).

3. Use of acylphloroglucinol heteroterpenoid compounds and pharmaceutically acceptable salts thereof in the preparation of drugs for resisting Gram-positive bacteria, characterized in that: The structural formula of the compound is shown in the following formula 1-2: Wherein, the Gram-positive bacteria is Methicillin-resistant Staphylococcus aureus (MRSA).

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