WO2019227969A1 - Ansamycin all-carbon cycle polyketide-type antibiotic and uses thereof in preparing antimicrobial medications or anti-tumor medications - Google Patents

Abstract

An ansamycin all-carbon cycle polyketide-type antibiotic and uses thereof in preparing antimicrobial medications or anti-tumor medications. Verrucosispora sp. SCSIO 07399 preserved on 22 October 2018 at Guangdong Microbial Culture Collection Center (GDMCC) having the address of: Guangdong Institute of Microbiology, 100 Xianlie Middle Road, Building 59, 5th floor, Guangzhou, and having the preservation number of GDMCC No. 60466. The fermentation culture of Verrucosispora sp. SCSIO 07399 can make new compounds kendomycins B to D, as shown in formulas (I) to (III). Said three compounds have markable antimicrobial and anti-tumor activities.

Description

Ansa full carbocyclic polyketide antibiotics and application thereof in preparing antibacterial or antitumor drugs Technical field:

The invention belongs to the technical field of microorganisms, and particularly relates to a marine wart spore (Verrucosispora sp.) SCSIO07399 and three kinds of new kendomycin compounds kendomycin BD extracted and isolated from the strain. The invention also relates to the preparation of the three new compounds by the strain Methods and applications.

Background technique:

Kendomycin is a polyketone compound of the ansamycin antibiotic family. The mother nucleus is characterized by a macrolide mixed with a quinone methyl chromophore. It was originally isolated from Streptomyces AL-71389 by Takeda Pharmaceutical Company as an antagonist of the endothelin receptor in 1996. [Bahnck KB, Rychnovsky SD. Formal synthesis of (-)-kendomycin featuring aprins-cyclization method to construct the macrocycle [J] .J Am Chem Soc, 2008.130 (39): 13177-13181]. In 1998, Su et al. Isolated it again from Streptomyces NRRL-21370, and the activity test results showed that it has an anti-osteoporosis effect [Xu S, Arimoto H. Strategies for construction, of the all-carbon macro-skeleton of the ansamycin [J] .J Antibioti (Tokyo), 2016, 47 (31): 203]. In 2000, Bode and Zeeck determined their absolute configuration through the Mosher reaction [Bode, H, B, Zeeck, A. Structure, and biosynthesis of Kendomycin, a carbocyclic, ansa-compound, from Streptomyces [J]. 58 (3): 323-328]. The biosynthetic pathway of Kendomycin antibiotics was first explained by Bode and Zeek using isotope labeling method [Bode, H, Zeeck, A. Biosynthesis of Kendomycin: Origin, of the oxygengen atomos, and Further Investment Investigations [J]. , 16 (16): 2665-2670]. Later, Wenzel et al. Cloned their biosynthetic gene clusters. Biosynthetic analysis results showed that the formation of the mother nucleus of this type of compound was responsible for the mixing of type I and type III PKS polyketide synthase, which was first reported in ansamycin antibiotics [Wenzel “SC , Bode, HB, Kochems, I, et al. A Type I / Type III polyketide synthase hybrids Biosynthetic pathway for the structurally unique unique compound [kendomycin] [Chem] Chembiochem, 2008, 9 (16): 271-2721]. According to reports, kendomycin antibiotics have good biological activity, not only a wide range of antibacterial spectrum, but also inhibit methicillin-resistant Staphylococcus aureus and vancomycin-resistant Staphylococcus aureus, and also have a significant number of human tumor cells. Toxic activity, and is expected to develop into endocortin receptor antagonists and anti-osteoporosis agents [Magauer T, Martin HJ, Mulzer J. Ring-closing metathesis and photo-fries reaction for the construction of the ansamycin antibioticskendomycin: development a protecting group free oxidation endgame [J] .Chemistry, 2010,16 (2): 507-519].

Summary of the invention:

One of the objectives of the present invention is to provide a marine wart spore (Verrucosispora sp.) SCSIO 07399 capable of producing kendomycin BD, which was deposited at the Guangdong Provincial Center for Microbial Strains (GDMCC) on October 22, 2018, address : 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Provincial Institute of Microbiology, Deposit No .: GDMCC No. 60466.

Another object of the present invention is to provide an ansa full carbocyclic polyketide antibiotic kendomycin B-D as shown in formulae (I), (II) and (III), or a pharmaceutically acceptable salt thereof:

Among them, formula (I) is kendomycin B, formula (II) is kendomycin C, and formula (III) is kendomycin D.

The third object of the present invention is to provide the application of marine wart spore (Verrucosispora sp.) SCSIO 07399 in the preparation of kendomycin B-D.

The fourth object of the present invention is to provide a method for preparing kendomycins B-D. The kendomycin B-D is prepared from a fermented culture of Verrucosispora sp. SCSIO 07399.

According to the present invention, preferably, kendomycin B-D is prepared from a fermented culture of Verrucosispora sp. SCSIO07399, and specifically includes the following steps:

(a) Prepare a fermented culture of Verrucosispora sp. SCSIO 07399, separate the fermentation supernatant and mycelium of the fermentation culture, extract the fermentation supernatant with methyl ethyl ketone, and concentrate the methyl ethyl ketone phase After that, extract A is obtained, and the mycelia are immersed and extracted with acetone, and the acetone extract is concentrated to obtain extract B;

(b) Combine extract A and extract B, and use silica gel column chromatography with chloroform-methanol as the eluent. The volume ratios are 100: 0, 99: 1, 98: 2, 96: 4, and 94: 6. Gradient elution was performed at 92,8, 9: 1, 8: 2, 5: 5, and 0: 100, and the chloroform-methanol volume ratios were collected at 92: 8, 9: 1, 8: 2, and 5: 5. Fractions Fr.A6 to A9, combined fractions Fr.A6 to A9, and subjected to reverse phase medium pressure liquid chromatography, eluting with a mobile phase CH ₃ CN / H ₂ O 10% to 100% v / v linear gradient for 120min, The flow rate is 10mL / min, and each fraction is 150mL. Eight fractions Fr.B1 to B8 are sequentially obtained. The fraction Fr.B5 is passed through a Sephex LH-20 glucan gel chromatography column, and the volume ratio of CHCl ₃ / MeOH is 1: 1. The phases were eluted, and the fractions were collected and purified to obtain kendomycin D. The fraction Fr.B7 was passed through a Sephex LH-20 glucan gel column and eluted with CHCl ₃ / MeOH volume ratio 1: 1 as the mobile phase. The fractions were collected. After further purification, kendomycin B and kendomycin C were obtained.

The fermented culture for the preparation of Verrucosispora sp. SCSIO 07399 is prepared by the following method:

Verrucosispora sp. SCSIO 07399 is inserted into a seed medium, and a seed culture liquid is obtained by fermentation. The seed culture liquid is connected to a fermentation medium, and a fermentation culture is obtained by fermentation. The formula of the seed medium For: 10g of soluble starch, 4g of yeast extract, 2g of bacteriological peptone and 30g of sea salt per liter, the balance is water; the formula of the fermentation medium is: 20g of soluble starch, 10g of glucose, 10g of maltose, 5g of corn flour per liter , 10g of malt extract powder, 2g of CaCO ₃ and 30g of sea salt, the balance is water.

A fifth object of the present invention is to provide the application of kendomycin B, kendomycin C, or kendomycin D, or a pharmaceutically acceptable salt thereof in the preparation of an antibacterial or antineoplastic drug.

A sixth object of the present invention is to provide an antibacterial or antineoplastic drug, which contains an effective amount of the kendomycin B, kendomycin C or kendomycin D, or a pharmaceutically acceptable salt thereof as an active ingredient.

Preferably, the antibacterial drug is a drug against Bacillus thuringiensis, Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, or methicillin-resistant Staphylococcus aureus; the antitumor drug is anti-human gastric cancer, human lung cancer , Human cervical cancer, human liver cancer, human breast adenocarcinoma, or human colorectal cancer.

The present invention provides a strain of marine wart spore (Verrucosispora sp.) SCSIO 07399 capable of producing kendomycin antibiotics. A new compound kendomycin BD can be prepared by using this bacterium. These three compounds have significant antibacterial and antitumor activities and have broad Application prospects.

The Verrucosispora sp. SCSIO07399 of the present invention was deposited on October 22, 2018 at the Guangdong Provincial Center for Microbial Strains (GDMCC), Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Institute of Microbiology, deposit number: GDMCC No. 60466.

Brief description of the drawings:

Fig. 1 is a morphological diagram of Verrucosispora sp. SCSIO 07399 on M-ISP4 medium;

Figure 2 is a phylogenetic tree of Verrucosispora sp. SCSIO 07399, where 07399 represents Verrucosis pora sp. SCSIO 07399;

Figure 3 is an HPLC chart of the production of kendomycin B and kendomycin C by SCSIO 07399 from Verrucosispora sp .;

Fig. 4 is an HPLC chart of kendomycinD produced by Verrucosispora sp. SCSIO 07399 fermentation.

Detailed ways:

The following examples are further illustrations of the present invention, but not limitation of the present invention.

Example 1: Isolation and identification of Verrucosispora sp. SCSIO 07399

The Verrucosispora sp. SCSIO 07399 of the present invention is isolated from a sediment sample in the South China Sea.

The taxonomic characteristics of this strain are:

1. Morphological characteristics:

The colonies are small, usually orange-yellow, slender mycelium, branched, and not broken. Basal hyphae were formed, and occasionally sparse slightly white aerial hyphae (as shown in Figure 1).

2. Molecular biology separation characteristics:

Extract the genomic DNA of the isolated strain, PCR-amplify its 16SrDNA sequence (the nucleotide sequence is shown in SEQ ID NO.1) by conventional methods, and perform sequencing analysis to construct a phylogenetic tree based on the 16SrDNA sequence (such as Figure 2) shows that the sequence similarity between strain SCSIO 07399 and Verrucosispora sp. R8-37 16SrDNA is 99%, indicating that strain SCSIO 07399 belongs to Verrucosispora sp.

In summary, the identified strain SCSIO 07399 belongs to a species of the genus Warthospora, named Verrucosispora sp. SCSIO 07399, which was deposited in the Guangdong Provincial Center for Microbial Strains on October 22, 2018. (GDMCC), Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Provincial Institute of Microbiology, Deposit No .: GDMCC No. 60466.

Example 2: Isolation and identification of kendomycin B-D

1. Preparation of fermented culture of Verrucosispora sp. SCSIO 07399:

(1) Preparation of seed medium and fermentation medium:

a) Preparation of seed medium: Each liter of seed medium contains 10g of soluble starch, 4g of yeast extract, 2g of bacteriological peptone, and 30g of sea salt, and the balance is water. The above components are mixed uniformly and the pH is adjusted to 7.0. Prepare 1L seed medium, and then aliquot it into 20 250mL Erlenmeyer flasks, each 50mL, and sterilize at 121 ° C for 30min.

b) Preparation of fermentation medium: Each liter of fermentation medium contains 20g of soluble starch, 10g of glucose, 10g of maltose, 5g of corn flour, 10g of malt extract powder, 2g of CaCO ₃ and 30g of sea salt. The balance is water. Mix well and adjust pH to 7.0. Prepare 1L fermentation medium, then aliquot it into 5 1L Erlenmeyer flasks and sterilize at 121 ° C for 30min for later use.

(2) Seed cultivation:

The activated marine wart spore (Verrucosispora sp.) SCSIO 07399 (deposited as: GDMCC No. 60466) was inserted into a 250 mL Erlenmeyer flask containing 50 mL of seed medium, and cultured on a shaker at 28 ° C and 200 rpm for 24 h To obtain a seed culture solution.

(3) Scale fermentation culture:

50 mL of the seed culture solution in the above Erlenmeyer flask was transferred to a 1 L Erlenmeyer flask containing 200 mL of fermentation medium, and cultured on a shaker at 28 ° C. and 200 r / min for 7 days to obtain Verrucosispora sp. ) Fermentation culture of SCSIO 07399.

2. Isolation of kendomycin B-D from Verrucosispora sp. SCSIO 07399

(1) Extraction of fermentation culture

The fermented culture of Verrucosispora sp. SCSIO 07399 was centrifuged (3800 ～ 4000r / min, 10 ～ 15min) to separate the fermentation supernatant from the mycelium; the fermentation supernatant was treated with methyl ethyl ketone, etc. Extraction was performed three times in volume. The methyl ethyl ketone phase was condensed by rotary evaporation to obtain extract A. The mycelium was immersed and extracted with acetone (2L), and the acetone extract was obtained by ultrasonic treatment. Cream B.

(2) Extraction of kendomycin B-D

By HPLC analysis, the components of extract A and extract B are similar. Extract A and extract B are combined, and subjected to normal phase silica gel column chromatography, using chloroform-methanol as the mobile phase. Different volume ratios (100: 0, 99 : 1, 98: 2, 96: 4, 94: 6, 92: 8, 9: 1, 8: 2, 5: 5, 0: 100), followed by gradient elution, and eluted under a gradient of 100% chloroform. The fraction was recorded as Fr.A1, the fraction eluted under a gradient of chloroform-methanol volume ratio of 99: 1 was recorded as Fr.A2, and the fraction eluted under a gradient of chloroform-methanol volume ratio of 98: 2 was recorded as Fr.A3. The fraction eluted under a gradient of chloroform-methanol volume ratio of 96: 4 is recorded as Fr.A4, and the fraction eluted under a gradient of chloroform-methanol volume ratio of 94: 6 is recorded as Fr.A5. The fraction eluted under a gradient of 92: 8 was recorded as Fr.A6, the fraction eluted under a gradient of 9: 1 was recorded as Fr.A7, and the volume ratio of chloroform-methanol was determined under a gradient of 8: 2. The eluted fraction was recorded as Fr.A8, the fraction eluted under a chloroform-methanol volume ratio of 5: 5 was recorded as Fr.A9, and the fraction eluted under a chloroform-methanol volume ratio was 0: 100 It is Fr.A10. The above-mentioned fractions Fr.A1 to A10 were analyzed by high performance liquid phase, and it was found that the fractions Fr.A6, Fr.A7, Fr.A8 and Fr.A9 contained kendomycin BD (1-3), and were combined. The combined fractions were subjected to reverse phase medium pressure liquid chromatography (CH ₃ CN / H ₂ O ₁₀ % to 100% v / v linear gradient elution 120min, flow rate 10mL / min), each fraction 150mL, 8 fractions Fr were sequentially obtained .B1 ～ B8. By HPLC, it was found that compound 3 was contained in Fr. B5, and compound 1 and compound 2 were contained in Fr. B7. Fractions Fr.B5 and Fr.B7 were purified by Sephex LH-20 dextran gel chromatography column (mobile phase CHCl ₃ / MeOH1 / 1v / v isocratic elution) to obtain Fr.C1 ～ C15 and Fr.D1 ～ D15. After HPLC detection, Fr.C5 ～ C9 (containing kendomycin D) were combined and separated and purified by reverse-phase ODS (YMC-Pack ODS-A column, 250 × 20mm, 5μm) (mobile phase CH ₃ CN / H ₂ O 55% to 100% v / v linear gradient elution for 30 min, flow rate 2.5 mL / min) (as shown in Figure 4), compound 3 (6.2 mg, retention time 16.2 min) was obtained, which is kendomycin D. Fr.D5 ～ D12 (containing kendomycin B and kendomycin C) were separated and purified by reverse-phase ODS (YMC-Pack ODS-A column, 250 × 20mm, 5μm) by semi-preparative high performance liquid chromatography (mobile phase CH ₃ CN / H ₂ O 70% ～ 100% v / v linear gradient elution for 30 min, flow rate 2.5 mL / min) (as shown in Figure 3), compound 1 (5.0 mg; retention time 26.0 min) and compound 2 (4.0 mg; retention Time 24.5min), compound 1 is kendomycin B, and compound 2 is kendomycin C.

Through structural analysis, the identification results of the compound 1-3 prepared from the fermentation culture of Verrucosispora sp. SCSIO 07399 of the present invention are as follows:

Compound 1 is a yellow powder with molecular formula C ₂₈ H ₄₀ O ₆ , HR-ESI-MS m / z 471.2757 ([MH] ^- ); [α] ²⁰ _D -59 (c 0.21, MeOH); UV (MeOH) λ _max (logε) 202 (4.276), 307 (3.681) nm; IR (film): v _max : 3325, 2937, 2358, 2341, 1602, 1456, 1359, 1195, 1095, 1022 cm ^-1 . In the high field of the ¹ H NMR spectrum, six methyl signals δ _H 0.73 (3H, d, J = 6.8 Hz, Me-26), δ _H 0.89 (3H, d, J = 6.6 Hz, Me-23), δ _H 0.91 (3H, d, J = 6.6 Hz, Me-21), δ _H 0.95 (3H, d, J = 7.0 Hz, Me-22), δ _H 0.95 (3H, d, J = 6.5 Hz, Me- 25) and δ _H 1.61 (3H, m, Me-24). Combining one-dimensional ¹³ C NMR, DEPT NMR and two-dimensional HSQC data, a total of 28 carbon signals are shown, including 6 methyl groups, 4 methylene groups, 12 methine groups, and 6 quaternary carbons according to the COSY and HMBC spectra The graph analysis can infer that C-5 / C-6 (C-21) / C-7 / C-8 (C-22) / C-9 / C-10 / C-11 / C-12 ( C-23) / C-13 and C-15 / C-16 (C-25) / C-17 / C-18 (C-26) fragments. H-12 and C-14 can be observed in the HMBC spectrum; H-13 and C-14, C-15, C-24; H-15 and C-13, C-14, C-24; H-16 Correlation with C-14 and H-24 with C-13, C-14, C-15, it is inferred that the above two fragments are connected through C-14. Combined with one-dimensional ¹³ C NMR and HMBC correlation, 8 typical aromatic carbons in compound 1 are shown: δ _C 173.9 (C-1), 96.7 (C-2), 183.9 (C-3), 148.2 (C-4) 113.0 (C-4a), 120.6 (C-19), 143.5 (C-20) and 131.4 (C-20a) are a polysubstituted pyranone ring structure. At the same time, through H-5 and C-4, C-4a, C-20a; H-6 and C-4a; H-17 and C-19; H-18 and C-19, C-20; H-26 Related to the HMBC of C-19, the planar structure of Compound 1 was determined.

Look up literature, NMR data and literature of compound 1 [Magauer T, Martin HJ, Mulzer J. Ring-closing metathesis and photo-fries reaction for the construction of the ansamycin antibiotic kendomycin: development of the protection of the group free free oxidation. Chemistry, 2010,16 (2): 507-519] Comparison of NMR data reported for kendomycin A, reported that compound 1 is a new compound of kendomycin antibiotics, named kendomycin B, whose structure is shown in formula (I), carbon spectrum And proton spectrum data are shown in Table 1.

Compound 2 is a yellow powder with molecular formula C ₂₉ H ₄₂ O ₆ S, HR-ESI-MS m / z 517.2633 ([MH] ^- ); [α] ²⁰ _D -547 (c 0.22, MeOH); UV (MeOH) λ _max (logε) 202 (4.265), 240 (3.490), 380 (3.600) nm; IR (film): v _max : 3298, 2937, 2358, 2333, 1597, 1421, 1008 cm ^-1 . By comparison of the ¹ H, ¹³ C and DEPT NMR spectra, Compound 2 is a homologue of 1. The difference is that there is one more -SCH ₃ structure than 2. Finally, according to the correlation between H-27 and C-20 in the HMBC spectrum, it was determined that the -SCH _{3 was} connected to C-20 and named kendomycin C. Its structure is shown in formula (II), and the carbon spectrum and hydrogen spectrum data are shown in Table 1. As shown.

Compound 3 is a yellow powder with molecular formula C ₃₃ H ₄₇ NO ₉ S, HR-ESI-MS m / z 471.2757 ([MH] ^- ); [α] ²⁰ _D -240 (c 0.34, MeOH); UV (MeOH) λ _max (logε) 202 (4.328), 244 (3.591), 382 (3.453) nm; IR (film): v _max : 3363, 2949, 2362, 2330, 1602, 1373, 1010 cm ^-1 . By nuclear magnetic spectrum assignment, it was determined that compound 3 and 1 contained the same 22-membered polysubstituted pyranolactone ring as the mother core structure. The main difference is that in the ¹ H NMR spectrum, there is an additional methyl signal δ _H 1.93 (3H, s, Me-31) in the high field region and δ _H 3.52 (1H, dd, J = 5.0, 13.6 Hz), δ _H 3.87 (1H, dd, J = 7.8, 13.6 Hz) and δ _H 4.60 (1H, dd, J = 5.0, 7.8 Hz) 3 signals; ⁵ C in ¹³ C NMR The signals include 1 methyl signal δ _C 22.7, 1 methylene signal δ _C 35.8, 1 methine signal, 1 carbonyl carbon δ _C 172.6 and 1 carboxyl carbon δ _C 173.9. H-27 (δ _H 3.52, δ _H 3.87) and C-20 (δ _C 151.5) and C-28 (δ _C 55.1H-28 (δ _H 4.60) and C-27 ( δ _C 35.8) C-29 (δ _C 173.9) and C-30 (δ _C 172.7), H-31 (δ _H 1.93) and C-30 (δ _C 172.7) are remotely correlated, and combine ¹ H- ¹ H The coupling of H-27 (δ _C 35.8) and H-28 (δ _H 4.60) in COSY infers the existence of acetylacetylcysteine. The position of the connection of acetylacetylcysteine is through H-27 in the HMBC spectrum (δ _H 3.52, δ _H 3.87) and C-20 (δ _C 151.5) were determined. The absolute configuration of azaacetylcysteine was determined chemically, and the L and D forms of acetylacetylcysteine and the base Compound 1 reacted under neutral conditions. Through analysis by HPLC and LC-MS, it was finally determined that the C-20 link in compound 3 was L-type acetylacetylcysteine. Therefore, the structure of compound 3 was finally determined and named kendomycin. D. Its structure is shown in formula (III), and its carbon spectrum and hydrogen spectrum data are shown in Table 1.

Table 1 Nuclear magnetic carbon spectrum and hydrogen spectrum data of Kendomycin B-D

^a Measured in CD ₃ OD.

The filter paper sheet method was used to test the antibacterial activity of the compound, and it was found that compounds 1-3 had certain antibacterial activity against most Gram-positive bacteria. Continue to select Bacillus thuringiensis BT01, Bacillus subtilis BS01, Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 292292, Staphyureococcus 745a And methicillin-resistant Staphylococcus aureus shhs-A1, 96-well plate method was used to determine the minimum inhibitory concentration, kanamycin, ampicillin and vancomycin were used as positive controls. The minimum inhibitory concentration (MIC) of compound 1 against Staphylococcus aureus ATCC 29213, Staphylococcus aureus 745524, Enterococcus faecalis ATCC 29212, and Bacillus thuringiensis BT01 were 0.5, 1.0, 2.0, 1.0 μg / mL, respectively, which were better than those of card. Positive control of natamycin, ampicillin and vancomycin, compound 2 against Staphylococcus aureus ATCC 29213, Staphylococcus aureus 745524, methicillin-resistant Staphylococcus aureus shhs-A1, Enterococcus faecalis ATCC 29292, and Su Yunjin The minimum inhibitory concentration (MIC) of Bacillus BT01 was 0.5, 1.0, 1.0, 0.5, 0.5 μg / mL, which was better than the positive controls of kanamycin, ampicillin, and vancomycin. Compound 3 against golden yellow grapes The minimum inhibitory concentration (MIC) of Coccus ATCC 29213, Staphylococcus aureus 745524 and Bacillus thuringiensis BT01 were 1.0, 2.0, 2.0 μg / mL, respectively, which were better than the positive controls of kanamycin, ampicillin and vancomycin. . The test results are shown in Table 2.

Table 2 MIC values of compounds 1-3 (μg / mL)

^a Vancomycin, kanamycin and ampicilin served as positive controls. The tests were performed in triplicate.

Compounds 1-3 were applied to human gastric cancer cell line MGC803 (human gastric cancer cell line), human lung cancer cell line A549 (human lung cancer cell line) and human cervical cancer cell line Hela (human cervical cancer cell line) by standard MTT method. Six human cancer cell lines including the human hepatocellular carcinoma cell line HepG2, the human breast adenocarcinoma cell line MCF-7, and the colorectal carcinoma cell line RKO Human liver cell line L02 (normal human human hepatic cell line) and human umbilical vein endothelial cell line Huvec-12 (normal human human vein endothelial cell line) were tested for cytotoxic activity, and cisplatin was used as a positive control. Compounds 1-3 all have significant antitumor activity. The test results are shown in Table 3. According to reports by Janssen et al., The cytotoxic activity of compounds 1 and 2 on A549 is significantly better than that of kendomycin A [Janssen CO, Lim, S, Lo EP, et al. Interaction of kendomycin and semi-synthetic analogues with the anti-apoptotic protein Bcl-xl [J]. Bioorg Med Chem Lett, 2008, 18 (21): 5771-5773].

Table 3 IC ₅₀ values (μM) of compounds 1-3

^a Data are expressed as mean of at least three independent experiments. Cis-platinum was used as positive control

Claims (10)

Verrucosispora sp. SCSIO 07399, whose deposit number is GDMCC No. 60466. Any of Ansa's full carbocyclic polyketide antibiotics or their pharmaceutically acceptable salts represented by formulae (I), (II) and (III):

Among them, formula (I) is kendomycin B, formula (II) is kendomycin C, and formula (III) is kendomycin D. The use of the marine wart spore (Verrucosispora sp.) SCSIO 07399 according to claim 1 in the preparation of kendomycin B, C or D according to claim 2. A method for preparing kendomycin BD according to claim 2, wherein the kendomycin BD is prepared from a fermented culture of Verrucosispora sp. SCSIO 07399 according to claim 1. . The method for preparing kendomycin B-D according to claim 4, characterized in that it specifically comprises the following steps: (a) Prepare a fermented culture of Verrucosispora sp. SCSIO 07399, separate the fermentation supernatant and mycelium of the fermentation culture, extract the fermentation supernatant with methyl ethyl ketone, and concentrate the methyl ethyl ketone phase After that, extract A is obtained, and the mycelia are immersed and extracted with acetone, and the acetone extract is concentrated to obtain extract B; (b) Combine extract A and extract B, and use silica gel column chromatography with chloroform-methanol as the eluent. The volume ratios are 100: 0, 99: 1, 98: 2, 96: 4, and 94: 6. Gradient elution was performed at 92,8, 9: 1, 8: 2, 5: 5, and 0: 100, and the chloroform-methanol volume ratios were collected at 92: 8, 9: 1, 8: 2, and 5: 5. Fractions Fr.A6 to A9, combined fractions Fr.A6 to A9, and subjected to reverse phase medium pressure liquid chromatography, eluting with a mobile phase CH ₃ CN / H ₂ O 10% to 100% v / v linear gradient for 120min, The flow rate is 10mL / min, and each fraction is 150mL. Eight fractions Fr.B1 to B8 are sequentially obtained. The fraction Fr.B5 is passed through a Sephex LH-20 glucan gel chromatography column, and the volume ratio of CHCl ₃ / MeOH is 1: 1. The phases were eluted, and the fractions were collected and purified to obtain kendomycin D. The fraction Fr.B7 was passed through a Sephex LH-20 glucan gel column and eluted with CHCl ₃ / MeOH volume ratio 1: 1 as the mobile phase. The fractions were collected. After further purification, kendomycin B and kendomycin C were obtained. The preparation method according to claim 5, wherein the fermentation culture for preparing Verrucosispora sp. SCSIO 07399 is prepared by the following method: Verrucosispora sp. SCSIO 07399 is inserted into a seed medium, and fermented to obtain a seed culture solution. The seed culture solution is connected to a fermentation medium, and a fermentation culture is obtained by fermentation. The formula is: 10g of soluble starch, 4g of yeast extract, 2g of bacteriological peptone and 30g of sea salt per liter, the balance is water; the formula of the fermentation medium is: 20g of soluble starch, 10g of glucose, 10g of maltose, corn flour per liter 5g, malt extract powder 10g, CaCO ₃ 2g and sea salt 30g, the balance is water. The use of kendomycin B, kendomycin C or kendomycin D, or a pharmaceutically acceptable salt thereof according to claim 2 in the preparation of an antibacterial or antineoplastic drug. The application according to claim 7, characterized in that the antibacterial drug is a drug against Bacillus thuringiensis, Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus or methicillin-resistant Staphylococcus aureus; said The anti-tumor drugs are drugs against human gastric cancer, human lung cancer, human cervical cancer, human liver cancer, human breast adenocarcinoma or human colorectal cancer. An antibacterial or antitumor drug, comprising an effective amount of kendomycin B, kendomycin C, or kendomycin D according to claim 2, or a pharmaceutically acceptable salt thereof as an active ingredient. The antibacterial or antitumor drug according to claim 9, wherein the antibacterial drug is anti-Bacillus thuringiensis, Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, or methicillin-resistant Staphylococcus aureus The antitumor drug is a drug against human gastric cancer, human lung cancer, human cervical cancer, human liver cancer, human breast adenocarcinoma or human colorectal cancer.

Images