CN104031052A - Antibiotic Indimicins A-E and preparation method thereof, and application of antibiotic Indimicins A-E in preparing antineoplastic drugs - Google Patents
Antibiotic Indimicins A-E and preparation method thereof, and application of antibiotic Indimicins A-E in preparing antineoplastic drugs Download PDFInfo
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Abstract
The invention discloses antibiotic Indimicins A-E and preparation method thereof and application in preparations of anti-tumor drugs. Indimicins A-E, shown in structural formula such as formula (I), it is to be fermented to generate by streptomycete Streptomyces sp.SCSIO03032, contain novelty 6 '-chloro- 1 ', 3 '-bis- -2 '-H- of methyl indole structures and to MCF-7 Human Breast Cancer Cells, human liver cancer cell HepG2, human lung cancer H460 cell and human glioma cell SF-268 have cytotoxic activity, and bioengineering transformation is carried out to it or modifying for chemical structure is expected to be developed into anti-cancer agent.
Description
Technical field:
The invention belongs to industrial microorganism field, be specifically related to new microbiotic Indimicins A – E and preparation method thereof and in the application of preparing in antitumor drug.
Background technology:
Indole alkaloid structure is various and have multiple biology, a pharmacologically active.At present, the analogue of some indole alkaloids as the inhibitor of protein kinase K or the inhibitor of DNA topoisomerase I for clinical trial (H.Nakano, and S.Omura, ' Chemical Biology of Natural Indolocarbazole Products:30Years since the Discovery of Staurosporine ', J Antibiot, 62 (2009), 17-26.).
Summary of the invention:
First object of the present invention is to provide five kinds of new indole alkaloid indimicins A – E with anti-tumor activity.
Five kinds of new indole alkaloid indimicins A – E of the present invention, its structure is suc as formula shown in (I):
Wherein work as R
1=CH
3, R
2when=H, be Indimicins A (1); Work as R
1=CH
3, R
2=CH
3time, be Indimicins B (2); Work as R
1=H, R
2=CH
3time, be Indimicins C (3); Work as R
1=H, R
2when=H, be Indimicins D (4); When in indimicin A, C-2 ' and C-2 " between carbon-carbon single bond while disconnecting, be indimicin E (5).
Second object of the present invention is to provide the preparation method of a kind of indole alkaloid Indimicins A – E.
Indimicins A – E of the present invention is that preparation separation obtains from the fermenting culture of streptomycete (Streptomyces sp.) SCSIO03032.
The present invention preferably prepares Indimicins A – E by the following method from the fermenting culture of streptomycete (Streptomyces sp.) SCSIO03032, and concrete steps are as follows:
The fermenting culture of a, preparation streptomycete (Streptomyces sp.) SCSIO03032, the fermented liquid of this fermenting culture and mycelium are separated, fermented liquid is through macroporous resin adsorption, after use acetone wash-out macroporous resin, after elutriant reclaims acetone, remaining water mixed liquid is extracted with ethyl acetate, and ethyl acetate layer obtains extractum A after distillation and concentration; Mycelium is first used acetone extraction, and after leaching liquid reclaims acetone, remaining water mixed liquid is extracted with ethyl acetate again, and ethyl acetate layer obtains medicinal extract B after distillation and concentration;
B, the crude extract that extractum A and medicinal extract B are merged is through silica gel column chromatography, by chloroform/methanol as eluent, carry out gradient elution from volume ratio 100:0～0:100, collect the cut Fr.2 that chloroform/methanol volume ratio 98:2 gradient elution gets off, by the anti-phase medium pressure liquid chromatography of ODS, water/methyl alcohol is as eluent, carry out gradient elution from volume ratio 100:0～0:100, collect the cut Fr.2-3 that water/methyl alcohol volume ratio 20:80 gradient elution gets off, after LH-20 gel column, using chloroform/methanol volume ratio 1:1 as moving phase wash-out, eluting fraction is again through thin-layer chromatography purifies and separates, obtain compound indimicin A, indimicin B, indimicin C, indimicin D and indimicin E.
The fermenting culture of preparation streptomycete (Streptomyces sp.) SCSIO03032 of described a) step is preparation by the following method preferably: streptomycete (Streptomyces sp.) SCSIO03032 of activation is accessed to seed culture medium, 28 DEG C, 200rpm, cultivate 48h and obtain seed liquor, seed liquor is linked in fermention medium with 10% inoculum size, 28 DEG C, 200rpm, shaking culture 120h, and make fermenting culture, described seed culture medium and the formula of fermention medium are all to contain in every liter of substratum: starch 10g, yeast powder 5g, peptone 4g, CaCO
32g, thick sea salt 30g, surplus is water, pH7.2.
The 3rd object of the present invention is to provide streptomycete (Streptomyces sp.) SCSIO03032 in the application of preparing in compound indimicin A, indimicin B, indimicin C, indimicin D or indimicin E.
The inventor found through experiments, compound indimicins A – E comprises mammary cancer MCF-7 to human cancer cell, people's liver cancer HepG2, people's lung cancer H460 and human glioma cell SF-268 have cytotoxic activity, and wherein indimicin B comprises the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 to human cancer cell
50respectively 10.0 μ M, 13.9 μ M, 19.1 μ M and 11.5 μ M; Indimicin A comprises the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 to human cancer cell
50respectively 23.7 μ M, 22.3 μ M, 41.2 μ M and 17.9 μ M; Indimicin C comprises the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 to human cancer cell
50respectively 21.8 μ M, 20.9 μ M, 29.7 μ M and 23.4 μ M; Indimicin D comprises the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 to human cancer cell
50respectively 22.4 μ M, 23.9 μ M, 21.5 μ M and 22.3 μ M; Indimicin E comprises the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 to human cancer cell
50respectively 36.4 μ M, 32.0 μ M, 43.7 μ M and 33.4 μ M.
Therefore, the 4th object of the present invention is to provide compound indimicin A, B, C, D or E in the application of preparing in antitumor drug.
Described antitumor drug is preferably anti-breast cancer, people's liver cancer, people's lung cancer or human glioma's medicine.
The 5th object of the present invention is to provide a kind of antitumor drug, it is characterized in that, indimicin A, the B, C, D or the E that comprise significant quantity are as active ingredient.
Described antitumor drug is preferably anti-breast cancer, people's liver cancer, people's lung cancer or human glioma's medicine.
The present invention separates and obtains 5 new indole alkaloid indimicins A – E with anti-tumor activity from streptomycete (Streptomyces sp.) SCSIO03032, it can be for the preparation of antitumor drug, and being expected exploitation by biotechnology or chemically modified becomes anti-cancer agent.
Streptomycete of the present invention (Streptomyces sp.) SCSIO03032 is preserved in Chinese Typical Representative culture collection center (CCTCC) on July 18th, 2011, address: Wuhan University of Wuhan, China city, its deposit number is CCTCC NO:M2011258, it is disclosed in the patent No.: ZL201210087537.7, denomination of invention is: in the patent of streptomycete, antineoplastic compound Spiro-Indimycin A-D and its preparation method and application.
Brief description of the drawings:
Fig. 1 is the extract-extractum A of supernatant liquor and the high-efficient liquid phase chromatogram of mycelial extract-medicinal extract B;
High performance liquid chromatography (HPLC) condition: chromatographic column is phenomex150 × 4.6mm (SphereClone SAX), moving phase comprises A phase and B phase, mobile phase A phase: the formic acid of acetonitrile+0.08% (volume fraction) of 10% (volume fraction), solvent is water, B phase flows: the acetonitrile of 90% (volume fraction), and solvent is water; Sample introduction program: 0-20min, mobile phase ratio is A phase/B phase (volume ratio): 95:5-0:100,20-21min, mobile phase ratio is A phase/B phase (volume ratio): 0:100,21-22min, mobile phase ratio is A phase/B phase (volume ratio): 0:100-95:5,22-30min, mobile phase ratio is A phase/B phase (volume ratio): 95:5, detects wavelength 254nm, flow velocity 1ml/min, wherein 1 representation compound 1,2 representation compound 2,3 representation compounds 3,4 representation compound 4,5 representation compounds 5.
The crucial HMBC of Fig. 2 compound 1,
1h–
1the crystalline diffraction structure of H COSY and compound 1 relevant with NOE.
The crucial HMBC of Fig. 3 compound 2-4,
1h–
1h COSY is relevant with NOE.
The CD collection of illustrative plates of Fig. 4 compound 2-4 in methanol solution.
The crucial HMBC of Fig. 5 compound 5 and
1h–
1h COSY is relevant, ECD collection of illustrative plates and (3 ' R)-5 of calculating and the ECD collection of illustrative plates of (3 ' S)-5 of compound 5 in methanol solution.
Embodiment:
Following examples are to further illustrate of the present invention, instead of limitation of the present invention.
Embodiment 1: separation and the preparation of active metabolite Indimicin A-E
1, substratum (seed culture medium and fermention medium): every liter contains starch 10g, yeast powder 5g, peptone 4g, CaCO
32g, sea salt 30g, surplus is water, pH7.2.121 DEG C, sterilizing 30min;
2, fermentation
2.1, seed culture: single bacterium colony of the streptomycete activating on culture dish (Streptomyces sp.) SCSIO03032 is accessed respectively to 18 bottles, in the taper culturing bottle of every bottle of 250mL that contains 50mL seed culture medium, 28 DEG C, 200rmin-1, cultivate 48h, make seed liquor 900mL.
2.2, fermentation culture: seed liquor is linked into 9L fermention medium with 10% inoculum size (volume percent) and (is placed in the taper culturing bottle of 250mL, every bottle contains 50ml fermention medium, totally 180 bottles) in, 28 DEG C, 200rmin-1, shaking culture 120h, obtains 9L fermenting culture.
3, extraction: fermenting culture first carries out centrifugation (3500rmin-1,8min), obtains supernatant liquor (fermented liquid) and the mycelium of 9L volume.Fermented liquid is through macroporous resin XAD16 Solid-Phase Extraction, after use acetone wash-out macroporous resin 3 times, after elutriant reclaims acetone, remaining water mixed liquid is extracted with ethyl acetate 3 times, and ethyl acetate layer obtains supernatant liquor extract-extractum A (3.6g) after distillation and concentration; At room temperature lixiviate of 2L acetone 3 times for mycelium, each 3 hours, united extraction liquid, remained the extraction of water mixed liquid 6L ethyl acetate after reclaim under reduced pressure acetone, and ethyl acetate layer underpressure distillation obtains mycelium extract-medicinal extract B (0.7g).
4, the extraction of active compound separates and qualification
The extraction of 4.1 Compound I ndimicin A-E separates and qualification: the sample of the extractum A that takes a morsel respectively and medicinal extract B is dissolved in 100 μ l methyl alcohol, centrifuging and taking supernatant 10 μ l, through HPLC sample detection, show, in extractum A, contain compound 1 (1), 4 (4) and 5 (5), in medicinal extract B, contain compound 2 (2) and 3 (3) (Fig. 1), extractum A and medicinal extract B are merged.By the crude extract of this merging through silicagel column (300-400 order) chromatography, by chloroform/methanol as eluent, carry out gradient elution from volume ratio 100:0～0:100, collect the cut Fr.2 (2.54g) that chloroform/methanol volume ratio 98:2 gradient elution gets off, evaporate to dryness, through the anti-phase medium pressure liquid chromatography of ODS (YMC*GEL ODS-A ball-shaped filling material, 120A aperture, 50um particle diameter, 240 × 4.0cm I.D.), flow velocity is 25ml/min, water/methyl alcohol is as eluent, carry out gradient elution from volume ratio 100:0～0:100, collect the cut Fr.2-3 (1.41g) that water/methyl alcohol volume ratio 20:80 gradient elution gets off, after LH-20 gel column (2.5*100cm), using chloroform/methanol volume ratio 1:1 as moving phase wash-out, every 50ml collects and merges into portion, order obtains Fr.2-3-A-F five duplicate samples, Fr.2-3-D (280mg) is prepared with thin-layer chromatography, collect thin layer prepare on plate with petrol ether/ethyl acetate volume ratio be the 9:1 cut that Rf value is 0.8 during as developping agent, be compound 1 (indimicin A) (100.6mg), Rf value is 0.7 cut, is compound 3 (indimicin C) (17.2mg), Rf value is 0.6 cut, is compound 5 (indimicin E) (53.5mg).Thin-layer chromatography preparation for Fr.2-3-C (25mg), collect on thin layer plate with petrol ether/ethyl acetate volume ratio be the 9:1 cut that Rf value is 0.8 during as developping agent, be compound 2 (indimicin B) (14.3mg).Fr.2-3-E (17mg) is prepared with thin-layer chromatography, collect on thin layer plate with petrol ether/ethyl acetate volume ratio be the 75:25 cut that Rf value is 0.8 during as developping agent, be compound 4 (indimicin D) (10.6mg).By structural analysis, 5 compound-compound 1-5 that prepare from the fermenting culture of streptomycete (Streptomyces sp.) SCSIO03032 of the present invention (corresponding compound indimycin A-E) (formula (I)) are identified it, and its qualification result is as follows:
Compound 1 (Indimicin A): white crystal,
uV (MeOH) λ
max(log ε) 252nm (4.55); 207nm (4.75); ECD (c3.9 × 10
-5m, MeOH) λ
max(Δ ε)-4.43 (251), 8.67 (227), 12.11 (217) nm; IR (KBr) ν
max3371,2291,1603,1477cm
-1;
1h NMR (500MHz, CDCl
3) and
13c NMR (125MHz, CDCl
3) data, in table 1, table 2; HRESIMS m/z[M-H]
-408.1051, (calcd for C
23h
20cl
2n
3, 408.1029).
The positive source high resolution electrospray ionization mass spectrum figure of compound 1 shows that its quasi-molecular ion peak is m/z408.1051[M+H]
+, infer that its molecular formula is C
23h
19cl
2n
3(calculated value is 408.1029, and degree of unsaturation is 15), IR composes at 3371cm
-1have strong absorption, there is NH in prompting in molecule.Analyze
1h,
13c and HSQC (table 1, table 2) data show that this compound contains three methyl [δ
h3.60 (3H, s, H-10), δ
c36.3; δ
h2.77 (3H, s, H-10 '), δ
c34.9; δ
h1.50 (3H, s, H-11 '), δ
c27.1], eight sp
2the methyne of hydridization, a sp
3methyne [the δ of hydridization
h4.02 (1H, s, H-2 '), δ
c72.8], 10 sp
2the quaternary carbon of hydridization and a sp
3quaternary carbon [the δ of hydridization
c45.5 (C-3 ')].Pass through
1h-
1the demonstration of H COSY atlas analysis contains two typical ABX coupling system [δ
h7.33 (1H, d, J=2.0Hz), 7.09 (1H, dd, J=8.5,2.0Hz), 6.43 (1H, d, J=8.5Hz) and δ
h7.71 (1H, d, J=2.0Hz), 7.18 (1H, dd, J=8.5,2.0Hz), 7.31 (1H, d, J=8.5Hz], in this explanation compound 1, contain two 1,2, the trisubstituted phenyl ring of 4-.These NMR data characteristicses of compound 1 and the spiroindimicins A-D of bibliographical information and lynamicins A-D (Zhang, W.; Liu, Z.; Li, S.; Yang, T.; Zhang, Q.; Ma, L.; Tian, X.; Zhang, H.; Huang, C.; Zhang, S.; Ju, J.; Shen, Y.; Zhang, C.Org.Lett.2012,14,3364-3367; McArthur, K.A.; Mitchell, S.S.; Tsueng, G.; Rheingold, A.; White, D.J.; Grodberg, J.; Lam, K.S.; Potts, B.C.J.Nat.Prod.2008,71,1732-1737) similar, prompting compound 1 belongs to the two indole ring alkaloid of single pyrroles family.Carefully analyze
1h-
1h COSY belonged in compound 16 " chloro-2 " relevant with HMBC, 3 " bis-replacement-1 "-H-indoles and 1-methyl-3, the two substituted azole rings (Fig. 2) of 4-.Different from compound lynamicins A-D is in compound 1, contain one rare 6 '-chloro-1 ', 3 '-bis-methyl-2 '-H-indoles, this is by H-10 ' (δ
h2.77) to C-2 ' (δ
c72.8) and C-9 ' (δ
c134.9), H-2 ' (δ
h4.02) to C-3 ' (δ
c45.5), and H-11 ' (δ
h1.50) to C-2 ' (δ
c72.8), C-3 ' (δ
c45.5) and C-4 ' (δ
c139.3) HMBC relevant (Fig. 2) is confirmed.Except this, " be connected to form six-ring, this is by H-2 ' (δ for C-2 ' and C-2 in compound 1
h4.02) to C-2 " (δ
c130.3), C-3 " (δ
c109.2) and C-4 (δ
c123.1) HMBC relevant (Fig. 2) confirms.Like this, determined the two dimensional structure of compound 1.H-2 ' and H in compound 1
3-11 ' cis-configuration be by H-2 ' and H
3-11 ' relevant (Fig. 2) determining of NOESY.The Cu target diffraction of compound 1 monocrystalline has further been confirmed its structure (Fig. 2), simultaneously according to the configuration of C-2 ' and C-3 ' in compound 1 being defined as to S and R (absolute configuration parameter-0.014 (12)) (Fig. 2).Therefore the structure of compound 1 suc as formula (I) 1 shown in, its R
1=CH
3, R
2=H, called after Indimicin A.
Compound 2 (Indimicin B): white solid,
uV (MeOH) λ
max(log ε) 207nm (4.81); ECD (c2.4 × 10
-5m, MeOH) λ
max(Δ ε)-7.81 (252), 5.94 (233), 34.23 (207) nm; IR (KBr) ν
max3339,2922,1600,1476cm
-1;
1h NMR (500MHz, CDCl
3) and
13c NMR (125MHz, CDCl
3) data are in table 1, table 2; HRESIMS m/z[M-H]
-422.1191, (calcd for C
23h
20cl
2n
3, 422.1185).
The positive source high resolution electrospray ionization mass spectrum figure of compound 2 shows that its quasi-molecular ion peak is m/z422.1191[M+H]
+, infer that its molecular formula is C
24h
21cl
2n
3(calculated value is 422.1185).Compound 2
1h and
13c spectrum data (table 1, table 2) are very similar to compound 1.Different places is many methyl in compound 2, from H
3-11 (δ
h2.60,3H, s) to C-2 (δ
c122.0) and C-3 (δ
c112.5) methyl is positioned C-2 position by HMBC.The two dimensional structure of compound 2 is further confirmed (Fig. 3) by 2DNMR collection of illustrative plates.Therefore the structure of compound 2 suc as formula (I) 2 shown in, its R
1=CH
3, R
2=CH
3, called after Indimicin B.
Compound 3 (Indimicin C): an off-white, optically inactive solid.
uV (MeOH) λ
max(log ε) 204nm (4.51); ECD (c2.4 × 10
-5m, MeOH) λ
max(Δ ε)-9.73 (254), 8.72 (235), 32.37 (208) nm; IR (KBr) ν
max3410,2921,1603,1479cm
-1;
1h NMR (500MHz, CDCl
3) and
13cNMR (125MHz, CDCl
3) data are in table 1, table 2; HRESIMS m/z[M-H]
-408.1039, (calcd for C
23h
20cl
2n
3, 408.10295).
Compound 3 is white solid.The positive source high resolution electrospray ionization mass spectrum figure of compound 3 shows that its quasi-molecular ion peak is m/z408.1039[M+H]
+, infer that its molecular formula is C
24h
21cl
2n
3(calculated value is 408.1029).Compound 3
1h and
13very similar (table 1, the table 2) of CNMR data and compound 2.According to the NH-1 (δ in compound 3
h7.641H, br s) and H-5 (δ
h6.341H, d, J=2.0Hz) between exist COSY relevant, infer that compound 3 lacks the methyl group of N-1 compared with compound 2.The two dimensional structure of compound 3 is further confirmed (Fig. 3) by 2D NMR collection of illustrative plates.Therefore the structure of compound 3 suc as formula (I) 3 shown in, its R
1=H, R
2=CH
3, called after Indimicin C.
Compound 4 (Indimicin D): white solid.
uV (MeOH) λ
max(log ε) 204nm (4.39); ECD (c2.6 × 10
-5m, MeOH) λ
max(Δ ε)-3.89 (251), 3.67 (228), 7.38 (207) nm; IR (KBr) ν
max3278,2924,1604,1481cm
-1;
1h NMR (500MHz, CDCl
3) and
13c NMR (125MHz, CDCl
3) data are in table 1, table 2; HRESIMS m/z[M-H]
-392.0712, (calcd for C
23h
20cl
2n
3, 392.0727).
Compound 4 is white solid, and the positive source high resolution electrospray ionization mass spectrum figure of compound 4 shows that its quasi-molecular ion peak is m/z392.0712[M+H]
+, infer that its molecular formula is C
22h
17cl
2n
3(calculated value is 392.0727).Compound 4
1h and
13cNMR spectrum data (table 1, table 2) are similar to compound 1.H-2 (δ in compound 4
h6.991H, dd, J=1.5,1.5)/NH-1 (δ
h8.041H, br s), with H-5 (δ
h6.531H, dd, J=1.5,1.5)/NH-1 (δ
h8.041H br is between s)
1h-
1hCOSY is relevant shows that compound 4 lacks N-1 position methyl group.The two dimensional structure of compound 4 is further confirmed (Fig. 3) by 2D NMR collection of illustrative plates.Identical with compound 1, compound 2 – 4 have two chiral carbon at C-2 ' and C-3 ' position, and their relative configuration is passed through H
3-11 ' with the relevant cis-configuration (Fig. 3) that is defined as of NOESY of between H-2 '.By their CD spectrum is compared (Fig. 4) with compound 1, the absolute configuration of C-2 ' and C-3 ' position is defined as S and R.Therefore the structure of compound 4 suc as formula (I) 4 shown in, its R
1=H, R
2=H, called after Indimicin D.
Compound 5 (Indimicin E): white solid.
uV (MeOH) λ
max(log ε) 203nm (4.50); ECD (c2.4 × 10
-5m, MeOH) λ
max(Δ ε)-4.24 (318), 12.98 (239), 13.80 (206) nm; IR (KBr) ν
max3418,2929,1722,1462cm
-1;
1h NMR (500MHz, CDCl
3) and
13c NMR (125MHz, CDCl
3) in table 1, table 2; HRESIMS m/z[M-H]
-410.1207, (calcd for C
23h
20cl
2n
3, 410.1185).
Compound 5 is white solid, and the positive source high resolution electrospray ionization mass spectrum figure of compound 5 shows that its quasi-molecular ion peak is m/z410.1207[M+H]
+, infer that its molecular formula is C
23h
21cl
2n
3(calculated value is 410.1185).Compound 5
1h,
13c and HSQC NMR data (table 1, table 2) show that it has similar structure fragment to compound 1: 6 " chloro-2 ", and 3 " bis-replacement-1 "-H-indoles, 6 '-chloro-1 ', 3 '-bis-methyl-2 '-H-indoles and 1-methyl-3, the two substituted azole rings of 4-.Different is in compound 5, to lack a methyne and a fragrant quaternary carbon contained in compound 1, many methylene radical [δ
h3.08,3.31, (2H, d, H-2 '); Two keys disconnect, this supposition by H3-10 ' in compound 5 (δ H2.32) to C-2 ' (δ C69.3) and H2-2 ' (δ H3.03,3.31) and H-2 relevant to the HMBC of C-3 ' (δ C43.7) " (δ H6.091H; s) " (δ H7.931H, the COSY (Fig. 5) of br between s) confirms br with NH-1.The CD spectrum that comparative compound 5 is measured in methanol solution and the ECD spectrum of the 3 ' R calculating and 3 ' S, find that the CD spectrum of measuring is consistent in the region of 200-400nm with the ECD spectrum of 3 ' R, all at 200-240, there is negative Ces in 270-350nm region, has positive Ces (Fig. 5) in 240-260nm region.Thus, deterministic compound 5 is at the R that is configured as of C-3 ' position.Therefore, the structure of compound 5 suc as formula (I) 5 shown in, called after Indimicin E, when in indimicin A, C-2 ' and C-2 " between carbon-carbon single bond while disconnecting, be indimicin E.
Table 1. Compound I ndimicins A – E's (1 – 5)
1h NMR (500MHz) nuclear magnetic data ownership
δ C69.3] and methyne [δ H6.09 (1H, s, a H-2 "); δ C125.4].Infer C-2 ' and C-2 in compound 5 " between carbon-to-carbon
Table 2. Compound I ndimicins A – E's (1 – 5)
13c NMR (125MHz) nuclear magnetic data ownership
The determination of cytotoxic activity of embodiment 2:Indimicins A-E
Indimicins A-E is for four kinds of tumor cell lines: MCF-7 Human Breast Cancer Cells, and human liver cancer cell HepG2, people's lung cancer H460 cell and human glioma cell SF-268 have carried out determination of activity, experimental technique reference [Wu, Z.C.; Li, D.L.; Chen, Y.C.; Zhang, W.M., A new isofuranonaphthalenone and benzopyrans from the endophytic fungus Nodulisporium sp.A4from Aquilaria sinensis.Helv.Chim.Acta2010,93, (5), 920-924.], (human cancer cell is comprised to the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 in contrast with cis-platinum
50respectively 6.0 μ M, 1.9 μ M, 3.6 μ M and 4.1 μ M).Result shows that indimicin B comprises the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 to human cancer cell
50respectively 10.0 μ M, 13.9 μ M, 19.1 μ M and 11.5 μ M; Indimicin A comprises the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 to human cancer cell
50respectively 23.7 μ M, 22.3 μ M, 41.2 μ M and 17.9 μ M; Indimicin C comprises the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 to human cancer cell
50respectively 21.8 μ M, 20.9 μ M, 29.7 μ M and 23.4 μ M; Indimicin D comprises the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 to human cancer cell
50respectively 22.4 μ M, 23.9 μ M, 21.5 μ M and 22.3 μ M; Indimicin E comprises the IC of mammary cancer MCF-7, people's lung cancer H460 cell, people's liver cancer HepG2 and human glioma cell SF-268 to human cancer cell
50respectively 36.4 μ M, 32.0 μ M, 43.7 μ M and 33.4 μ M.Indimicins A-E has cytotoxic activity to the performance of four kinds of surveyed cells, and being expected exploitation by biotechnology or chemically modified becomes anti-cancer agent.
Claims (9)
1. compound indimicins A-E, its structure is suc as formula shown in (I):
Wherein, compound 1 is indimicin A, its R
1=CH
3, R
2=H; Compound 2 is indimicin B, its R
1=CH
3, R
2=CH
3; Compound 3 is indimicin C, its R
1=H, R
2=CH
3; Compound 4 is indimicin D, its R
1=H, R
2=H; Compound 5 is indimicin E.
2. the preparation method of a compound indimicins A-E claimed in claim 1, it is characterized in that, described compound indimicins A-E is that preparation separation obtains from the fermenting culture of streptomycete (Streptomyces sp.) SCSIO03032.
3. preparation method according to claim 2, is characterized in that, concrete steps are as follows:
The fermenting culture of a, preparation streptomycete (Streptomyces sp.) SCSIO03032, the fermented liquid of this fermenting culture and mycelium are separated, fermented liquid is through macroporous resin adsorption, after use acetone wash-out macroporous resin, after elutriant reclaims acetone, remaining water mixed liquid is extracted with ethyl acetate, and ethyl acetate layer obtains extractum A after distillation and concentration; Mycelium is first used acetone extraction, and after leaching liquid reclaims acetone, remaining water mixed liquid is extracted with ethyl acetate again, and ethyl acetate layer obtains medicinal extract B after distillation and concentration;
B, the crude extract that extractum A and medicinal extract B are merged is through silica gel column chromatography, by chloroform/methanol as eluent, carry out gradient elution from volume ratio 100:0～0:100, collect the cut Fr.2 that chloroform/methanol volume ratio 98:2 gradient elution gets off, by the anti-phase medium pressure liquid chromatography of ODS, water/methyl alcohol is as eluent, carry out gradient elution from volume ratio 100:0～0:100, collect the cut Fr.2-3 that water/methyl alcohol volume ratio 20:80 gradient elution gets off, after LH-20 gel column, using chloroform/methanol volume ratio 1:1 as moving phase wash-out, eluting fraction is again through thin-layer chromatography purifies and separates, obtain compound indimicin A, indimicin B, indimicin C, indimicin D and indimicin E.
4. preparation method according to claim 3, it is characterized in that, the fermenting culture of preparation streptomycete (Streptomyces sp.) SCSIO03032 of described a step is to prepare by the following method: streptomycete (Streptomyces sp.) SCSIO03032 of activation is accessed to seed culture medium, 28 DEG C, 200rpm, cultivate 48h and obtain seed liquor, seed liquor is linked in fermention medium with 10% inoculum size, 28 DEG C, 200rpm, shaking culture 120h, and make fermenting culture, described seed culture medium and the formula of fermention medium are all to contain in every liter of substratum: starch 10g, yeast powder 5g, peptone 4g, CaCO32g, thick sea salt 30g, surplus is water, pH7.2.
5. the application of streptomycete (Streptomyces sp.) SCSIO03032 in preparation compound indimicin A claimed in claim 1, indimicin B, indimicin C, indimicin D or indimicin E.
6. compound indimicins A claimed in claim 1, B, C, D or E are in the application of preparing in antitumor drug.
7. application according to claim 6, is characterized in that, described antitumor drug is anti-breast cancer, people's liver cancer, people's lung cancer or human glioma's medicine.
8. an antitumor drug, is characterized in that, indimicin A, the B, C, D or the E that comprise significant quantity are as active ingredient.
9. antitumor drug according to claim 8, is characterized in that, described antitumor drug is anti-breast cancer, people's liver cancer, people's lung cancer or human glioma's medicine.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104928305A (en) * | 2015-07-09 | 2015-09-23 | 中国科学院南海海洋研究所 | Biosynthetic gene cluster of big-ring lactam compound heronamides and application thereof |
| CN105112322A (en) * | 2015-08-07 | 2015-12-02 | 浙江大学 | Grisic quinone A, grisic quinone B, and preparation method and medical application of grisic quinone A and grisic quinone B |
| CN116354899A (en) * | 2023-03-03 | 2023-06-30 | 西北农林科技大学 | Hybrid polyketone compound containing oxazole ring, and preparation method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001044247A2 (en) * | 1999-12-16 | 2001-06-21 | Eli Lilly And Company | Agents and methods for the treatment of proliferative diseases |
| CN1303859A (en) * | 1999-11-17 | 2001-07-18 | 阿迪尔公司 | Carbazole derivatives their preparing process and pharmaceutical composition containing them |
| WO2002030941A2 (en) * | 2000-10-06 | 2002-04-18 | Bristol-Myers Squibb Company | Topoisomerase inhibitors |
| CN1571793A (en) * | 2001-10-22 | 2005-01-26 | 瑟维尔实验室 | Novel hydroxyalkyl indolocarbazole derivatives, preparation method and pharmaceutical compositions containing same |
| CN1852914A (en) * | 2003-09-16 | 2006-10-25 | 万有制药株式会社 | Novel indolopyrrolocarbazole derivative with antitumor activity |
-
2014
- 2014-05-20 CN CN201410214540.XA patent/CN104031052B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1303859A (en) * | 1999-11-17 | 2001-07-18 | 阿迪尔公司 | Carbazole derivatives their preparing process and pharmaceutical composition containing them |
| WO2001044247A2 (en) * | 1999-12-16 | 2001-06-21 | Eli Lilly And Company | Agents and methods for the treatment of proliferative diseases |
| WO2002030941A2 (en) * | 2000-10-06 | 2002-04-18 | Bristol-Myers Squibb Company | Topoisomerase inhibitors |
| CN1571793A (en) * | 2001-10-22 | 2005-01-26 | 瑟维尔实验室 | Novel hydroxyalkyl indolocarbazole derivatives, preparation method and pharmaceutical compositions containing same |
| CN1852914A (en) * | 2003-09-16 | 2006-10-25 | 万有制药株式会社 | Novel indolopyrrolocarbazole derivative with antitumor activity |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104928305A (en) * | 2015-07-09 | 2015-09-23 | 中国科学院南海海洋研究所 | Biosynthetic gene cluster of big-ring lactam compound heronamides and application thereof |
| CN104928305B (en) * | 2015-07-09 | 2019-06-11 | 中国科学院南海海洋研究所 | The biological synthesis gene cluster of macrolactams compound heronamides a kind of and its application |
| CN105112322A (en) * | 2015-08-07 | 2015-12-02 | 浙江大学 | Grisic quinone A, grisic quinone B, and preparation method and medical application of grisic quinone A and grisic quinone B |
| CN105112322B (en) * | 2015-08-07 | 2018-07-06 | 浙江大学 | Grey mold quinone A and B and preparation method thereof and medical usage |
| CN116354899A (en) * | 2023-03-03 | 2023-06-30 | 西北农林科技大学 | Hybrid polyketone compound containing oxazole ring, and preparation method and application thereof |
| CN116354899B (en) * | 2023-03-03 | 2024-02-27 | 西北农林科技大学 | Hybrid polyketone compound containing oxazole ring, and preparation method and application thereof |
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