CN108383815B - Polyketone compounds, preparation method and application thereof - Google Patents

Polyketone compounds, preparation method and application thereof Download PDF

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CN108383815B
CN108383815B CN201810309651.7A CN201810309651A CN108383815B CN 108383815 B CN108383815 B CN 108383815B CN 201810309651 A CN201810309651 A CN 201810309651A CN 108383815 B CN108383815 B CN 108383815B
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张怡轩
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Abstract

The invention relates to the field of medicines, in particular to a preparation method and medical application of polyketone compounds. The invention provides a series of polyketide compounds shown in a general formula (I) and pharmaceutically acceptable salts, solvates or isomers thereof, wherein the compounds are obtained by separating and extracting from fermentation cultures of fasciolopsis fungi (Cephalotrichum sp.). The compounds have good anti-tumor activity. Meanwhile, the compounds also have certain activity for resisting human pathogenic drug-Resistant bacteria MRSA (Methicillin-Resistant Staphylococcus aureus) and VRE (Vancomycin-Resistant Enterococcus).

Description

Polyketone compounds, preparation method and application thereof
Technical Field
The invention belongs to the technical field of medicines, relates to a polyketone compound, and a preparation method and application thereof, and particularly relates to application of the polyketone compound in preparation of anti-tumor and anti-drug-resistant bacteria medicines.
Background
Tumor (tumor) refers to a new organism (neograwth) formed by local tissue cell proliferation of the body under the action of various tumorigenic factors, because the new organism is mostly in the form of space-occupying block-shaped protrusion, also called neoplasms (neoplasms).
According to the cellular characteristics of the new organism and the degree of harm to the organism, tumors are divided into two major categories, namely benign tumors and malignant tumors, and cancers are a general term for malignant tumors.
In 2017, in 2 months, the national cancer center releases the latest cancer data in China, and summarizes the data of 347 national cancer registration points. The latest published is the morbidity and mortality data of 2013. About 1 million people diagnose cancer per day and about 7 people diagnose cancer per minute.
Therefore, the discovery and development of new antitumor drugs is urgent, the screening of antitumor compounds is an important field of new drug development, and the discovery of specific compounds capable of killing tumor cells will have important significance for human life health.
Methicillin-resistant staphylococcus aureus (MRSA) is a pathogenic bacterium with a wide epidemic range, strong pathogenicity, and high morbidity and mortality, and has resistance to various antibiotics. Therefore, the search for novel MRSA-resistant lead compounds and the development of novel drug-resistant bacteria-resistant drugs become one of the hotspots in the current drug research and development.
Vancomycin-Resistant enterococci (VRE for short), also known as Vancomycin-Resistant enterococci, is a bacterium belonging to the genus Enterococcus. Enterococci (Enterococcus) are known to have persistent resistance, and the early resistance to Penicillin (Penicillin) and the recent resistance to vancomycin (vancomycin) make the clinical treatment of enterococci difficult, so that the search for more effective drugs for antagonizing VRE is important.
Fungi is a treasure pool endowed to human by nature, and a large amount of active substances are extracted from secondary metabolites of the fungi and are used for treating diseases caused by various types of tumor cells and various pathogenic bacteria. The extraction of bioactive components using fungal metabolites is becoming increasingly appreciated by those skilled in the relevant art.
Disclosure of Invention
The invention aims to provide a series of polyketone compounds.
The second purpose of the invention is to provide the method for extracting, separating and purifying the polyketone compounds.
The third purpose of the invention is to provide a pharmaceutical composition containing the polyketide.
The fourth purpose of the invention is to provide the application of the polyketide compound or the pharmaceutical composition in preparing anti-tumor and anti-drug-resistant bacteria drugs.
The invention is realized by the following technical scheme:
the invention provides polyketone compounds shown as a general formula (I) and pharmaceutically acceptable salts, solvates or isomers thereof:
Figure BDA0001621765730000021
wherein:
R1、R4、R5is H, C1-C4Carbonyl group, C2-C4Alkenyl, hydroxy, substituted or unsubstituted C1-C10Alkyl, wherein the substituent is hydroxyl, halogen, amino or cyano;
R2is OH, OAc, O (CH)2)mCOCH3、(CH2)mOCOCH2(CHOH)nCH3Substituted or unsubstituted C1-C6The substituent is hydroxyl, halogen, amino and cyano, wherein m is 0-10, and n is 0-10;
R3is H, Ac, substituted or unsubstituted C1-C10Alkyl, wherein the substituent is hydroxyl, halogen, amino or cyano;
the invention preferably selects polyketone compounds shown in a general formula (I) and pharmaceutically acceptable salts, solvates or isomers thereof:
wherein:
R1、R4、R5is H, C1-C4Carbonyl group, C2-C4Alkenyl, hydroxy, substituted or unsubstituted C1-C4Alkyl, the substituent is hydroxyl;
R2is OH, OAc, (CH)2)mOCOCH2(CHOH)nCH3Substituted or unsubstituted C1-C6The substituent is hydroxyl, halogen, amino and cyano, wherein m is 0-4, and n is 0-4;
R3h, Ac;
the invention preferably selects polyketone compounds shown in a general formula (I) and pharmaceutically acceptable salts, solvates or isomers thereof:
wherein:
R1、R4、R5is H, C1-C4Carbonyl group, C2-C3Alkenyl, hydroxy, substituted or unsubstituted C1-C4Alkyl, the substituent is hydroxyl;
R2is OH, OAc, OCO (CH)2)m(CHOH)nCH3Wherein m is 0-4, n is 0-4;
R3h, Ac;
the invention preferably selects polyketone compounds shown in a general formula (I) and pharmaceutically acceptable salts, solvates or isomers thereof:
wherein:
R1、R4、R5is H, C1-C2Carbonyl group, C2-C3Alkenyl, hydroxy, substituted or unsubstituted C1-C4Alkyl, the substituent is hydroxyl;
R2is OH, OAc, OCO (CH)2)m(CHOH)nCH3Wherein m is 0 and n is 0;
R3h, Ac;
the invention preferably selects polyketone compounds shown in a general formula (I) and pharmaceutically acceptable salts, solvates or isomers thereof:
wherein:
R1、R4、R5is H, C1-C2Carbonyl group, C2-C3Alkenyl, hydroxy, substituted or unsubstituted C1-C4Alkyl, the substituent is hydroxyl;
R2is OH, OAc, OCO (CH)2)m(CHOH)nCH3Wherein, in the step (A),m＝1，n＝1；
R3h, Ac;
the following polyketone compounds and pharmaceutically acceptable salts, solvates or isomers thereof are preferred in the present invention:
Figure BDA0001621765730000041
Figure BDA0001621765730000051
the invention also provides a preparation method of the polyketone compound, which comprises the following steps:
(1) fermentation production of strains:
selecting a solid rice culture medium, diluting a strain cercospora cephamii (Cephalotrichum sp) with sterile water, uniformly inoculating the strain cercospora cephamii on the rice culture medium, and culturing at the constant temperature of 25-35 ℃ for 25-40 days to obtain a fermentation product.
(2) Preparing a total extract:
and (2) soaking the fermentation product obtained in the step (1) by using ethyl acetate, performing ultrasonic treatment, and then performing reduced pressure distillation to obtain a total extract.
(3) Separation and purification of compounds:
and (3) roughly separating the total extract in the step (2) into three sections by silica gel column chromatography by using a dichloromethane/trichloromethane-methanol system (V/V is 80:1-3: 1):
the first section is a crude fraction of dichloromethane/trichloromethane-methanol (V/V) ═ 60:1-40:1, methanol-water (10-100% methanol) is used as a mobile phase, ODS column chromatography separation is firstly carried out, then 60-70% methanol elution fraction obtained by ODS column separation is separated, methanol is used as a mobile phase, Sephadex LH-20 separation is carried out, all fractions are obtained, and finally, a double-wavelength full-preparation liquid phase is used for separation and purification by using methanol-water (60-65% methanol) as a mobile phase, so that the compound Z1 is obtained.
The second stage is that dichloromethane/trichloromethane-methanol (V/V) ═ 80:1-60:1 and crude fractions of 40:1-20:1 are combined, petroleum ether-acetone (V/V) ═ 7:1, 5:1, 3:1) is used as a mobile phase, silica gel column separation is firstly carried out, then a dual-wavelength full-preparation liquid phase is utilized, methanol-water or acetonitrile-water is used as the mobile phase, 275nm is used as a detection wavelength, and 20-25% acetonitrile-water is used as the mobile phase for preparing the liquid phase, so that compounds Z2, Z3 and Z7 are obtained; using 272nm as a detection wavelength and methanol-water (V/V is 73-77: 37-33) as a mobile phase for preparing a liquid phase to obtain a compound Z4; using 272nm as a detection wavelength and methanol-water (V/V is 68-72: 32-38) as a mobile phase for preparing a liquid phase to obtain compounds Z5 and Z6; the conditions for preparing the thin-layer plate developing agent are as follows: preparing a compound Z9 from methanol 15: 1-20: 1; thus, 7 compounds were obtained.
The third stage is a crude fraction of dichloromethane/chloroform-methanol (V/V) ═ 10:1-3:1, silica gel column separation is carried out by taking petroleum ether-acetone (V/V ═ 4:1, 2:1, 1:1) as a mobile phase, and then compound Z8 is obtained by utilizing a two-wavelength full-preparation liquid phase, acetonitrile-water as a mobile phase, 268nm as a detection wavelength, and 30% -35% acetonitrile-water as a mobile phase of the preparation liquid phase.
The preparation method of the polyketone compound comprises the following steps:
the solid rice culture medium in the step (1) comprises the following components: 40g of rice and 50-70 mL of water.
The inoculation concentration of the strain Isaria xiaojianensis (Cephalotrichum sp.) in the step (1) is as follows: spore concentration of 10 in 1mL7～109(cfu/mL)。
The soaking time in the step (2) is as follows: 3-12 h; ultrasonic conditions are as follows: 2-5 min.
The double wavelength in the step (3) refers to: the detector for preparing the liquid phase is an ultraviolet dual-wavelength detector, and corresponding wavelengths can be set according to the ultraviolet absorption characteristics of the compound.
In the silica gel column chromatography of the step (3), the gradient of dichloromethane/trichloromethane-methanol is as follows: V/V is 80:1, 50:1, 30:1, 20:1, 10:1, 5:1, 3: 1.
The invention also relates to a pharmaceutical composition which comprises the polyketide compound and pharmaceutically acceptable salts, solvates or isomers thereof and a pharmaceutically acceptable carrier.
The invention provides application of polyketone compounds, pharmaceutically acceptable salts, solvates or isomers thereof or pharmaceutical compositions thereof in preparing antitumor drugs. Pharmacological experiments prove that the compound separated and identified by the invention has stronger antitumor effect on human cervical cancer HeLa cells, human lung cancer A549 cells, human kidney cancer 786-O cells, human liver cancer HepG2, human breast cancer MCF-7 cells and human breast cancer MDA-MB231, and the effect is better than that of positive control drug 5-fluorouracil, especially the effect of the compound Z1-Z6 is more obvious.
The invention also provides application of the polyketone compound, pharmaceutically acceptable salt, solvate or isomer thereof or a pharmaceutical composition thereof in preparation of drugs for resisting drug-resistant bacteria. The drug-resistant bacteria are methicillin-resistant staphylococcus aureus (MRSA) and vancomycin-resistant enterococcus (VRE).
The compound Z1 of the invention has stronger anti-drug-resistant bacterial activity, wherein the compound has anti-MRSA IC5010ug/mL with activity against VRE and an IC50 of 25 ug/mL.
The fascicularis microsporus (Cephalotrichum sp.) used by the invention is preserved in China general microbiological culture Collection center (CGMCC), and the address is as follows: the microbiological research institute of western road No.1, 3, national academy of sciences, north-kyo, the rising area, the preservation date: 29/01/2018, accession number: CGMCC No. 15382.
Detailed Description
Example 1
The enrichment and preparation of polyketone compounds comprises the following steps:
(1) fermentation production of strains: selecting solid rice culture medium (250ml triangular flask, 40g rice 53ml water), diluting the strain (Cephalotrichum sp.) with sterile water, uniformly inoculating on the rice culture medium, and culturing at constant temperature of 30 deg.C for 35-40 days.
(2) Preparing an extract: soaking the fermented product in ethyl acetate, performing ultrasonic treatment for 3 times to obtain total components, and distilling under reduced pressure to obtain total extract.
(3) Separation and purification of compounds: fermenting to obtain total extract, and performing silica gel column chromatography with dichloromethane-methanol system to obtain three sections. The first stage is a crude fraction of dichloromethane-methanol (V/V) ═ 50:1, methanol-water is taken as a mobile phase, ODS column chromatography is firstly carried out, then the fraction obtained by ODS column chromatography is separated, methanol is taken as the mobile phase, Sephadex LH-20 is used for separation, and finally a double-wavelength full-preparation liquid phase is used for separation and purification by taking methanol-water as the mobile phase to obtain a compound Z1. The second stage was dichloromethane-methanol (V/V) ═ 80:1, 30: 1. the crude fraction of 20:1 is separated by silica gel column with petroleum ether-acetone (V/V is 7:1, 5:1 and 3:1) as a mobile phase, then a dual-wavelength full-preparation liquid phase is utilized, and fractions obtained by the silica gel column are sequentially prepared with methanol-water (V/V is 55-75: 45-25) as a mobile phase, so that 7 compounds of corresponding compounds Z2, Z3, Z4, Z5, Z6, Z7 and Z9 are obtained. The third stage is a crude fraction of dichloromethane-methanol (V/V) ═ 10:1, 5:1, 3:1:1, which is subjected to silica gel column separation using petroleum ether-acetone as a mobile phase, and the fraction obtained from the silica gel column is prepared using a two-wavelength full-preparative liquid phase using methanol-water as a mobile phase, to obtain compound Z8.
Nuclear magnetic data for the compounds are as follows:
z1: is a light yellow powder, HRESIMS shows the peak of the excimer ion M/z [ M + Na ]]+457.2729 molecular formula C25H38O6And the molecular weight is 434.2729.
1H NMR(600MHz CDCL3-d)δ:6.17(1H,d,9.6,H-3),7.00(1H,dd,9.6,6.0,H-4),5.31(1H,dd,6.0,24.,H-5),4.09(1H,dd,9,2.4,H-6),2.14(1H,m,H-7),1.04(1H,ddd,13.5,9.9,4.8,H-8α),1.18(1H,ddd,13.5,9.3,5.1,H-8β),1.65(1H,m,H-9),1.44(1H,dd,13.3,9.9,H-10α),2.06(1H,brd,13.1,H-10β),5.09(1H,m,H-12),1.51(3H,d,6.6,H-13),1.12(3H,d,6.5,H-14),0.74(3H,d,6.6,H-15),1.49(3H,s,H-16),5.70(1H,d,15.7,H-2′),7.30(1H,d,15.7,H-3′),5.70(1H,brd,9.3,H-5′),2.87(1H,m,H-6′),1.65(1H,m,H-7′α),1.79(1H,m,H-7′β),3.57(1H,m,H-8′α)3.71(1H,m,H-8′β),1.81(3H,s,H-9′),3.59(2H,d,6.1,H-10′)。
13C NMR(150MHz，CDCL3-d)δ:163.3(C-2),124.9(C-3),140.5(C-4),61.7(C-5),83.3(C-6),31.4(C-7),39.9(C-8),27.9(C-9),46.3(C-10),134.2(C-11),120.0(C-12),13.3(C-13),15.8(C-14),20.6(C-15),15.5(C-16),166.0(C-1′),115.0(C-2′),150.9(C-3′),134.7(C-4′),143.2(C-5′),39.2(C-6′),34.7(C-7′),60.6(C-8′),12.6(C-9′),65.8(C-10′)。
Z2: as a pale yellow oil, HRESIMS showed an excimer peak m/z 503.2414(calcd fo r 503.2412, C)25H40ClO8) Molecular formula C25H40O8。
1H NMR(600MHz CDCL3-d)δ:6.22(1H,d,9.6,H-3),7.03(1H,dd,9.6,6.0,H-4),5.45(1H,dd,6.0,2.4,H-5),4.14(1H,dd,9.6,1.8,H-6),2.13(1H,m,H-7),1.52(1H,m,H-8α),0.98(1H,m,H-8β),1.79(1H,m,H-9),1.39(1H,m,H-10α),0.96(1H,m,H-10β),3.53(1H,m,H-12),1.06(3H,d,6.6,H-13),1.17(3H,d,6.6,H-14),0.99(3H,d,6.6,H-15),1.12(3H,s,H-16),5.85(1H,d,15.6,H-2′),7.37(1H,d,15.6,H-3′),5.82(1H,d,15.6,H-5′),2.88(1H,m,H-6′),1.80(1H,m,H-7′α),1.61(1H,m,H-7′β),3.71(1H,m,H-8′α)3.62(1H,m,H-8′β),1.84(3H,s,H-9′),3.58(2H,m,H-10′)。
13C NMR(150MHz，CDCL3-d)δ:163.4(C-2),125.1(C-3),140.6(C-4),61.4(C-5),83.5(C-6),31.7(C-7),40.8(C-8),25.7(C-9),41.8(C-10),74.9(C-11),74.9(C-12),17.4(C-13),16.5(C-14),24.1(C-15),24.0(C-16),166.3(C-1′),115.2(C-2′),151.1(C-3′),134.8(C-4′),143.8(C-5′),39.3(C-6′),34.6(C-7′),60.7(C-8′),12.7(C-9′),65.9(C-10′)。
Z3: as an opalescent powder, HRESIMS showed the excimer peak m/z 445.2197, (calcd for 445.2202, C)23H34NaO7) Molecular formula is C23H34O7。
1H NMR(CDCL3-d，600MHz)δ:6.22(1H,d,9.6,H-3),7.04(1H,dd,9.6,6.0,H-4),5.42(1H,dd,6.0,2.4,H-5),4.17(1H,dd,9.0,2.4,H-6),2.04(1H,m,H-7),1.31(1H,m,H-8α),1.06(1H,m,H-8β),2.7(1H,m,H-9),2.39(1H,m,H-10α),2.07(1H,m,H-10β),2.08(1H,s,H-12),1.15(3H,d,6.6,H-13),0.91(3H,d,6.6,H-14),5.83(1H,d,15.6,H-2′),7.37(1H,d,15.6,H-3′),5.81(1H,d,9.6,H-5′),2.89(1H,m,H-6′),1.80(1H,m,H-7′α)1.62(m,H-7′β),3.74(1H,m,H-8′α)3.62(1H,m,H-8′β),1.85(3H,s,H-9′),3.61(2H,m,H-10′)。
13C NMR(CDCL3D, 150MHz) delta 163.2(C-2),125.1(C-3),140.5(C-4),61.7(C-5),82.9(C-6),31.7(C-7),39.6(C-8),30.4(C-9),49.9(C-10),208.5(C-11),26.5(C-12),16.1(C-13),21.2(C-14),166.1(C-1 '), 115.0 (C-2'), 151.2(C-3 '), 134.6 (C-4'), 143.8(C-5 '), 39.3 (C-6'), 34.8(C-7 '), 60.7 (C-8'), 12.7(C-9 '), 65.9 (C-10'). Z4: a pale yellow oil, ESIMS, showed the excimer peak M/z 499.5[ M + Na ]]+Molecular formula is C27H40O7Molecular weight is 476.2.
1H NMR(600MHz CDCL3-d)δ:6.21(1H,d,9.6,H-3),7.04(1H,dd,9.6,6.1,H-4),5.35(1H,dd,5.9,2.4,H-5),4.12(1H,brd,8.8,H-6),2.16(1H,m,H-7),1.05(1H,m,H-8α)1.23(1H,m,H-8β),1.67(1H,m,H-9),1.43(1H,m,H-10α)2.06(1H,m,H-10β),5.11(1H,q,6.0,H-12),1.54(3H,d,6.3,H-13),1.04(3H,d,6.3,H-14),0.84(3H,d,6.3,H-15),1.52(3H,s,H-16),5.81(1H,d,15.7,H-2′),7.34(1H,d,15.7,H-3′),5.71(1H,brd,9.9,H-5′),2.87(1H,m,H-6′),1.59(1H,m,H-7′α)1.89(1H,m,H-7′β),3.95(1H,m,H-8′α)4.10(m,H-8′β),1.81(3H,s,H-9′),3.56(1H,m,H-10′α)3.62(1H,m,H-10′β),2.02(3H,s,8′-OCO 3CH)。
13C NMR(150MHz，CDCL3-d)δ:163.5(C-2),125.1(C-3),140.7(C-4),61.9(C-5),83.4(C-6),31.5(C-7),40.1(C-8),27.9(C-9),46.4(C-10),134.3(C-11),120.2(C-12),13.5(C-13),15.7(C-14),20.7(C-15),15.7(C-16),166.2(C-1′),115.5(C-2′),150.8(C-3′),135.7(C-4′),142.8(C-5′),38.9(C-6′),30.4(C-7′),62.4(C-8′),12.8(C-9′),65.9(C-10′),21.1(8′-OCOCH3),171.1(8′-OCOCH3)。
Z5, light yellow oil, ESIMS shows the excimer peak M/Z [ M + Na ]]+499.6 molecular formula C27H40O7Molecular weight is 476.2.
1H NMR(CDCL3-d，600MHz)δ：6.22(1H,d,9.6,H-3),7.04(1H,dd,9.6,6.0,H-4),5.38(1H,dd,6.01,2.45,H-5),4.12(1H,brd,8.8,H-6),2.14(1H,m,H-7),1.06(1H,m,H-8α)1.20(1H,m,H-8β),1.71(1H,m,H-9),1.43(1H,m,H-10α)2.07(1H,m,H-10β),5.11(1H,q,6.0,H-12),1.54(3H,d,6.3,H-13),1.15(3H,d,6.3,H-14),0.79(3H,d,6.3,H-15),1.52(3H,s,H-16),5.81(1H,d,15.7,H-2′),7.33(1H,d,15.7,H-3′),5.79(1H,d,9.8,H-5′),3.05(1H,m,H-6′),1.57(1H,m,H-7′)1.79(1H,m,H-7′),3.57(1H,m,H-8′α)3.67(m,H-8′β),1.82(3H,s,H-9′),4.03(2H,m,H-10′)2.02(3H,s,10′-OCO 3CH)。
13C NMR(CDCL3-d，150MHz)δ:163.5(C-2),125.2(C-3),140.8(C-4),61.9(C-5),83.5(C-6),31.6(C-7),40.1(C-8),28.1(C-9),45.9(C-10),134.4(C-11),120.3(C-12),13.6(C-13),15.8(C-14),21.1(C-15),16.1(C-16),166.3(C-1′),115.5(C-2′),151.0(C-3′),135.4(C-4′),142.6(C-5′),35.2(C-6′),34.5(C-7′),60.4(C-8′),12.8(C-9′),67.0(C-10′),20.8(10′-OCOCH3),171.2(10′-OCOCH3)。
Z6: a pale yellow oil, ESIMS, showed the excimer peak M/z 543.4[ M + Na ]]+Molecular formula is C29H44O8And the molecular weight is 520.
1H NMR(CDCL3-d，600MHz)δ:6.22(1H,d,9.6,H-3),7.04(1H,9.6,6.0,H-4),5.36(1H,dd,6.0,2.4,H-5),4.14(1H,dd,8.4,1.8,H-6),2.17(1H,m,H-7),1.21(1H,m,H-8α)1.04(1H,m,H-8β),1.68(1H,m,H-9),2.05(1H,m,H-10α)1.44(1H,dd,13.2,10.2,H-10β),5.12(1H,dd,12.6,6.0,H-12),1.55(3H,d,6.6,H-13),1.15(3H,d,6.6,H-14),0.78(3H,d,6.6,H-15),1.53(3H,s,H-16),5.82(1H,d,15.6,H-2′),7.35(1H,d,15.6,H-3′),5.72(1H,d,9.6,H-5′),2.84(1H,m,H-6′),1.94(1H,m,H-7′α)1.61(1H,m,H-7′β),4.17(1H,m,H-8′α)4.01(1H,m,H-8′β),1.82(3H,s,H-9′),3.63(1H,H-10′α)3.57(1H,m,H-10′β),2.44(2H,m,H-2″),4.19(1H,m,H-3″),1.22(3H,d,6.2,H-4″)。
13C NMR(CDCL3-d,150MHz)δ：163.3(C-2),125.0(C-3),140.6(C-4),61.7(C-5),83.2(C-6),31.3(C-7),39.9(C-8),27.8(C-9),46.3(C-10),134.9(C-11),120.1(C-12),13.3(C-13),15.8(C-14),20.6(C-15),15.5(C-16),166.0(C-1′),115.4(C-2′),150.6(C-3′),135.5(C-4′),142.4(C-5′),38.7(C-6′),30.2(C-7′),62.5(C-8′),12.7(C-9′),65.6(C-10′),172.8(C-1″),42.8(C-2″),64.2(C-3″),22.5(C-4″)。
Z7: ESIMS showed the excimer peak m/z503.2421(calcd for 503.2412, C) as a pale yellow oil25H40ClO8).-Molecular formula is C25H40O8。
1H NMR(600MHz CDCL3-d)δ:6.24(1H,d,9.6,H-3),7.01(1H,dd,9.6,6.0,H-4),5.49(1H,dd,6.0,2.4,H-5),4.13(1H,dd,9.6,1.8,H-6),2..05(1H,m,H-7),1.23(1H,m,H-8α),1.09(1H,m,H-8β),1.79(1H,m,H-9),1.23(1H,m,H-10α),0.99(1H,m,H-10β),3.44(1H,m,H-12),1.03(3H,d,6.6,H-13),1.17(3H,d,6.6,H-14),1.05(3H,d,5.4,H-15),1.12(3H,s,H-16),5.81(1H,m,15.6,H-2′),7.37(1H,d,15.6,H-3′),5.82(1H,d,15.6,H-5′),2.90(1H,m,H-6′),1.80(1H,m,H-7′α),1.59(1H,m,H-7′β),3.71(1H,m,H-8′α)3.60(1H,m,H-8′β),1.84(3H,s,H-9′),3.56(1H,m,H-10′)3.64(1H,m,H-10′)。
13C NMR(150MHz，CDCL3-d)δ:163.3(C-2),125.2(C-3),140.4(C-4),61.1(C-5),83.4(C-6),31.4(C-7),39.9(C-8),25.9(C-9),41.3(C-10),75.3(C-11),75.2(C-12),17.5(C-13),16.0(C-14),23.5(C-15),23.4(C-16),166.1(C-1′),114.9(C-2′),151.4(C-3′),134.9(C-4′),144.3(C-5′),39.3(C-6′),34.6(C-7′),60.6(C-8′),12.8(C-9′),66.0(C-10′)。
Z8: is light yellow oil with molecular formula C25H38O7Molecular weight 450.2.
1H NMR(600MHz CDCL3-d)δ:6.17(1H,d,9.6,H-3),6.95(1H,dd,9.6,6.0,H-4),5.37(1H,dd,6.0,2.5,H-5),4.07(1H,dd,9.5,2.5,H-6),2.10(1H,m,H-7),1.01(1H,m,H-8α)1.20(1H,m,H-8β),1.72(1H,m,H-9),1.53(1H,dd,14.9,10.2,H-10α)2.10(1H,dd,14.9,3.5,H-10β),4.12(1H,q,6.5,H-12),1.10(3H,d,6.5,H-13),1.15(3H,d,6.5,H-14),0.82(3H,d,6.5,H-15),4.69(1H,s,H-16),5.01(1H,s,H-16),5.75(1H,d,15.8,-2'),7.30(1H,d,15.7,H-3'),5.73(1H,d,10.2,H-5'),2.81(1H,m,H-6'),1.59(2H,dd,13.9,8.2,H-7'α)1.79(2H,dd,13.9,8.1,H-7'β),3.54(1H,m,H-8'α),3.64(1H,m,H-8'β),1.77(3H,s,H-9'),3.57(1H,dd,10.7,6.5,H-10'α),3.63(1H,dd,10.7,5.8,H-10'β)。
13C NMR(150MHz，CDCL3-d)δ:163.4(C-2),125.2(C-3),140.5(C-4),61.5(C-5),83.4(C-6),31.7(C-7),40.1(C-8),28.6(C-9),38.8(C-10),151.5(C-11),70.8(C-12),22.4(C-13),16.4(C-14),21.0(C-15),109.6(C-16),166.3(C-1'),115.1(C-2'),151.5(C-3'),134.9(C-4'),144.0(C-5'),39.4(C-6'),34.7(C-7'),60.7(C-8'),12.8(C-9'),66.0(C-10')。
Z9: ESIMS showed the peak of the excimer ion M/z 459.0[ M + Na ] as a milky white powder]+Molecular formula is C25H40O6And the molecular weight is 436.2.
1H NMR(600MHz CDCL3-d)δ:2.49(2H,m,H-3),1.94(1H,m,H-4α)2.29(1H,m,H-4β),4.75(1H,m,H-5),4.97(1H,dd,4.1,5.8,H-6),2.02(1H,m,H-7),1.0(1H,m,H-8α)1.35(1H,m,H-8β),1.71(1H,m,H-9),1.63(1H,m,H-10α)2.02(1H,m,H-10β),5.16(1H,q,6.3,H-12),1.55(3H,d,6.7,H-13),0.95(3H,d,6.7,H-14),0.81(3H,d,6.6,H-15),1.55(3H,s,H-16),5.86(1H,d,15.7,H-2′),7.35(1H,d,15.7,H-3′),5.77(1H,d,9.9,H-5′),2.89(1H,m,H-6′),1.82(1H,m,H-7′α)1.63(1H,m,H-7′β),3.61(1H,m,H-8′α)3.75(1H,m,H-8′β),1.86(3H,s,H-9′),3.61(2H,m,H-10′)
13C NMR(150MHz，CDCL3-d)δ:176.9(C-2),28.2(C-3),24.4(C-4),79.3(C-5),76.9(C-6),31.7(C-7),40.7(C-8),28.2(C-9),47.1(C-10),134.3(C-11),120.0(C-12),13.3(C-13),15.5(C-14),20.3(C-15),15.7(C-16),166.7(C-1′),115.8(C-2′),150.2(C-3′),134.7(C-4′),142.7(C-5′),39.2(C-6′),34.8(C-7′),60.6(C-8′),12.7(C-9′),65.8(C-10′)
Example 2
Adopting an MTT reduction method, purchasing human cervical cancer HeLa cells, human lung cancer A549 cells, human breast cancer MCF-7 cells, human breast cancer MDA-MB231 and human liver cancer HepG2 cells from American Type Culture Collection (ATCC, Rockville, Md., USA); the human kidney cancer 786-O cells are purchased from cell banks of Chinese academy of sciences (Shanghai, China), the cells are adherent cells, and adherent tumor cells in logarithmic growth phase are selectedSelecting adherent tumor cells in logarithmic growth phase, digesting with pancreatin, and preparing into 5 × 10 with culture medium containing 10% fetal calf serum4Per mL of cell suspension, seeded in 96-well plates at 100 uL/well, 37 ℃ 5% CO2And culturing for 24 h. The experimental group is replaced by new culture solution containing samples to be tested with different concentrations, the control group is replaced by culture solution containing equal volume of solvent, each group is provided with 3 parallel holes, the temperature is 37 ℃, and the concentration is 5% CO2Culturing for 48 h. The supernatant was discarded, carefully washed 2 times with PBS, 100uL of freshly prepared medium containing 0.5mg/mL MTT was added to each well, and incubation continued at 37 ℃ for 4 h. The supernatant was carefully discarded, 150uL of DMSO was added, and after mixing for 10min with a micro-shaker, the optical density was measured at 492nm using a microplate reader. 5-fluorouracil (5-FU) is selected as a positive control drug.
Tumor cell growth inhibition (%) was [ a492 (negative control) -a 492 (plus drug group) ]/a492 (negative control) × 100%
Results of in vitro antitumor Activity of a Compound
Figure BDA0001621765730000131
786-5-Fu is a positive control drug, ND means no measurement 786-O human renal cancer cells; HeLa human cervical cancer cells; a549 human lung cancer cells; MCF-7 human breast cancer cells; MDA-MB231 human breast cancer cells; HepG2 human hepatoma cells.
Example 3
And (3) detection of antibacterial activity:
(1) diluting the test bacteria: detecting activation of MRSA and VRE strains, diluting with sterile water to ultraviolet OD600nm is 0.3 (10)8cfu/mL), diluting the above concentration by 100 times, adding into LB culture medium, and mixing.
(2) Preparing a liquid medicine: various compounds to be tested were dissolved and diluted with DMSO.
(3) Inoculation experiment:
experimental groups: 198uL test bacterial suspension +2uL drug sample (Z1)
Blank control group: 198uL culture medium +2uL drug sample (Z1)
Solvent control group: 198uL Medium +2uL DMSO
Positive control group: 198uL test bacterial suspension +2uL positive control drug (commercial gentamicin/vancomycin concentration)
Negative control group: 198uL bacterial suspension for detection
Each group of experiments is simultaneously provided with 3 parallel experiments
(4) Culturing: the 96-well plate is placed in an incubator at 37 ℃ for constant-temperature culture for 12h, and the OD value is measured at 600nm by using a microplate reader.
The bacteriostatic rate (%) was (1-A/B). times.100%
Average OD value of experimental group-blank control group OD value
Mean OD value of solvent control group-blank medium control group OD value
IC of two different compounds against drug-resistant bacteria50(ug/mL)
Compound (I) MRSA VRE
Z1 10 25
Z2 50 30
Z3 40 35
Z4 20 >100
Z5 15 80
Z6 25 >100
Z7 50 40
Z8 >100 >100
Z9 80 >100
As can be seen from Table two, the various compounds have different anti-drug-resistant bacteria activities, wherein the activity of Z1 is strongest, and the anti-drug-resistant bacteria MRSA IC thereof5010 ug/mL; IC against drug-resistant bacteria VRE50＝25ug/mL。

Claims (9)

1. Polyketides of general formula (I) and pharmaceutically acceptable salts thereof,
Figure 347448DEST_PATH_IMAGE001
wherein:
R1、R4、R5is H, hydroxy, substituted or unsubstituted C1-C10Alkyl, said substituent being hydroxyA group;
R2is substituted or unsubstituted C1-C6Alkyl, the substituent is hydroxyl;
R3is H.
2. Polyketone compounds and pharmaceutically acceptable salts thereof,
Figure 336132DEST_PATH_IMAGE002
Figure 619346DEST_PATH_IMAGE003
Figure 35284DEST_PATH_IMAGE004
。
3. the preparation method of the polyketone compound comprises the following steps:
(1) fermentation production of strains:
selecting a solid rice culture medium, diluting a strain of Cercospora oligospora (Cephalotrichum microsporum) with the preservation number of CGMCC No.15382 with sterile water, uniformly inoculating the diluted strain on the rice culture medium, and culturing at the constant temperature of 25-35 ℃ for 25-40 days to obtain a fermented product;
(2) preparing a total extract:
soaking the fermentation product obtained in the step (1) by using ethyl acetate, performing ultrasonic treatment, and then performing reduced pressure distillation to obtain a total extract;
(3) separation and purification of compounds:
and (3) roughly separating the total extract in the step (2) into three sections by using a silica gel column chromatography with a dichloromethane/trichloromethane-methanol system with the volume ratio of 80:1-3: 1:
the first section is a crude fraction of dichloromethane/trichloromethane-methanol with a volume ratio of 60:1-40:1, ODS column chromatography is firstly carried out by taking 10-100% methanol-water as a mobile phase, then 60-70% methanol elution fraction obtained by ODS column separation is separated by taking methanol as the mobile phase, Sephadex LH-20 separation is carried out to obtain all fractions, and finally, a double-wavelength full-preparation liquid phase is used for separation and purification by taking 60-65% methanol as the mobile phase to obtain a compound Z1;
in the second stage, dichloromethane/chloroform-methanol with the volume ratio of 80:1-60:1 and crude fraction with the volume ratio of 40:1-20:1 are combined, petroleum ether-acetone with the volume ratio of 7:1, 5:1 and 3:1 is taken as a mobile phase, silica gel column separation is firstly carried out, then a dual-wavelength full-preparation liquid phase is utilized, and methanol-water or acetonitrile-water is taken as the mobile phase to prepare compounds Z2-Z7 and Z9;
the third section is a crude fraction of dichloromethane/trichloromethane-methanol with the volume ratio of 10:1-3:1, and the crude fraction is subjected to silica gel column separation by taking petroleum ether-acetone with the volume ratio of 4:1, 2:1 and 1:1 as a mobile phase, and then a compound Z8 is prepared by utilizing a double-wavelength full-preparation liquid phase and taking 30-35% acetonitrile-water as a mobile phase;
Figure 771159DEST_PATH_IMAGE005
Figure 297955DEST_PATH_IMAGE006
Figure 701255DEST_PATH_IMAGE007
Figure 40093DEST_PATH_IMAGE008
Figure 263264DEST_PATH_IMAGE009
Figure 593751DEST_PATH_IMAGE010
Figure DEST_PATH_IMAGE011
Figure 179453DEST_PATH_IMAGE012
Figure 874876DEST_PATH_IMAGE013
。
4. the method according to claim 3,
in the second stage of preparation in the step (3), compounds Z2, Z3 and Z7 are obtained by taking 275nm as a detection wavelength and 20-25% acetonitrile-water as a mobile phase of the preparation liquid phase; using 272nm as a detection wavelength and methanol-water with a volume ratio of 73-77: 37-33 as a mobile phase for preparing a liquid phase to obtain a compound Z4; taking 272nm as a detection wavelength, and taking methanol-water with a volume ratio of 68-72: 32-38 as a mobile phase for preparing a liquid phase to obtain compounds Z5 and Z6; the conditions for preparing the thin-layer plate developing agent are as follows: preparing a compound Z9 from methanol =15: 1-20: 1; thus, 7 compounds were obtained.
5. A pharmaceutical composition comprising a polyketide compound as claimed in claim 1 or 2 and a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
6. Use of a polyketide compound as claimed in claim 1 or claim 2 and a pharmaceutically acceptable salt thereof or a pharmaceutical composition as claimed in claim 5 in the preparation of an anti-tumour medicament.
7. Use of a polyketide compound of claim 1 or 2 and a pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 5 in the preparation of a medicament against drug-resistant bacteria MRSA or VRE.
8. The application of the following compounds in preparing the drug-resistant bacteria MRSA medicine:
Figure DEST_PATH_IMAGE014
Figure 178819DEST_PATH_IMAGE008
Figure 922784DEST_PATH_IMAGE015
Figure 159730DEST_PATH_IMAGE016
。
9. the application of the following compounds in preparing the drug-resistant bacteria VRE drugs:
Figure 698159DEST_PATH_IMAGE014
Figure 20556DEST_PATH_IMAGE017
。
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