Polyene macrocyclic lactone compound, preparation method and application thereof

Abstract

The application belongs to the technical field of microorganisms and novel food and medicine, and particularly relates to a novel polyene macrocyclic lactam compound and application thereof, and further discloses a method for preparing the polyene macrocyclic lactam compound based on a fermentation method. The polyene macrocyclic lactam compounds FW8-1 and FW8-4 both have polyene macrocyclic lactam structures, and due to the addition reaction in the molecule, the novel structure of intramolecular double ring is formed, which provides a compound reserve for researching and developing new potential anti-tumor, anti-inflammatory, anti-angiogenesis, anti-fibrosis, blood pressure regulation, immunoregulation and neuroprotective drugs.

Description

Technical Field

[0001] This invention belongs to the field of microbial and novel food and pharmaceutical technology, specifically relating to a novel polyene macrocyclic lactam compound and its applications, and further disclosing a method for preparing the polyene macrocyclic lactam compound based on fermentation. Background Technology

[0002] Polyene macrolide antibiotics are a class of natural compounds containing macrolide structures, primarily produced by the metabolism of some actinomycetes. These compounds are named for their long olefinic chains with multiple conjugated double bonds. Their activity is mainly manifested in their potent antibiotic effects, especially their inhibitory effects on fungi. Examples include nystatin, currently used to treat Candida infections; amphotericin B, used to treat severe fungal infections of Aspergillus, Candida, and Cryptococcus; and natamycin, widely used in the pharmaceutical and food industries to prevent the growth of molds and yeasts. However, issues such as drug solubility and high toxicity limit the clinical use of these drugs. Furthermore, the emergence of drug-resistant bacteria has prompted researchers to continuously search for new polyene compounds or their derivatives in hopes of finding antibiotics with lower toxicity and a broader antibiotic spectrum. In addition, polyene macrolides, due to their specific structural characteristics, have also shown potential medical applications in antiviral and anticancer fields, but these applications are still in the research and exploration stage.

[0003] For example, Chinese patent CN107287131A discloses a novel 26-membered polyene macrocyclic lactam compound, FW05328-1, isolated from the microbial secondary metabolites of marine micromonas FIM05-328. This compound exhibits significant activity against human esophageal squamous cell carcinoma cell lines, but it is unstable and easily degraded. Another example is Chinese patent CN1112939865A, which discloses a novel homologue of FIM05-328, FW05328-d, isolated from micromonas FIM MA181224. This homologue exhibits slightly better stability and activity against mouse melanoma cell lines, human esophageal cancer cell lines, and human colon cancer cell lines, but its antitumor activity is significantly reduced.

[0004] Therefore, the search for novel structures and the study of the activity of polyene macrocyclic lactam structural analogs have positive guiding significance for subsequent structural chemical modifications. Summary of the Invention

[0005] Therefore, the technical problem to be solved by the present invention is to provide a novel polyene macrocyclic lactam compound;

[0006] The second technical problem to be solved by the present invention is to provide a method for preparing the polyene macrocyclic lactam compound based on fermentation;

[0007] The third technical problem to be solved by the present invention is to provide the use of the polyene macrocyclic lactam compound.

[0008] To solve the above-mentioned technical problems, the present invention provides a polyene macrocyclic lactam compound, wherein the polyene macrocyclic lactam compound comprises compound FW8-1 represented by formula (I) and / or compound FW8-4 represented by formula (II):

[0009]

[0010] The present invention also discloses a method for preparing the polyene macrocyclic lactam compound by fermentation, comprising the step of inoculating marine micromonospora into a suitable fermentation medium for fermentation culture;

[0011] The marine micromonospora strain, classified as Micromonospora sp. FIM-MA181224, has been deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.17139.

[0012] Specifically, the method for preparing the polyene macrocyclic lactam compound by fermentation includes the following steps:

[0013] (1) Seed culture: The marine micromonospora sp. FIM-MA181224 preserved on slant was inoculated into liquid seed culture medium and cultured at constant temperature. The seed culture was collected and used for later use.

[0014] (2) Fermentation broth culture: The seed liquid is transferred to the fermentation medium for constant temperature culture to obtain the fermentation product containing the desired polyene macrocyclic lactam compound.

[0015] Specifically, in the method for preparing the polyene macrocyclic lactam compound by fermentation, in step (1), the liquid seed culture medium comprises the following components in the indicated mass percentages: soluble starch 1.5-2.0%, glucose 0.3-0.5%, peptone 0.3-0.5%, yeast extract 0.3-0.5%, MgSO4·7H2O 0.01-0.05%, NaCl 0.05-0.1%, (NH4)2SO4 0.01-0.05%, CaCO3 0.05-0.1%, pH 6.0-8.0;

[0016] Preferably, the temperature of the isothermal incubation step is 25-35℃, and the incubation time is 1-3 days.

[0017] Specifically, in the method for preparing the polyene macrocyclic lactam compound by fermentation, in step (2), the fermentation culture medium comprises the following components in the indicated mass percentages: soluble starch 1.6-2.0%, glucose 0.3-0.6%, peptone 0.4-0.6%, soybean meal 0.2-0.3%, yeast powder 0.15-0.25%, K2HPO4·3H2O 0.03-0.06%, MgSO4·7H2O 0.01-0.05%, NaCl 0.05-0.1%, (NH4)2SO4 0.03-0.06%, CaCO3 0.08-0.12%, sea salt 0.8-0.12%, pH 6.2-8.0;

[0018] Preferably, the temperature for the isothermal incubation step is 25-35℃, and the incubation is carried out with shaking for 3-6 days.

[0019] Specifically, the method for preparing the polyene macrocyclic lactam compound by fermentation further includes the steps of extraction and purification of the polyene macrocyclic lactam compound, specifically including:

[0020] (3) Extraction: The collected fermentation products are separated into fermentation broth and mycelium respectively; the fermentation broth is added to macroporous resin for adsorption, and desorbed by ethanol solution to collect crude extract A; the mycelium is extracted by alcohol solvent, and the extract is collected and concentrated to obtain crude extract B;

[0021] (4) Purification: Take the crude extract A and crude extract B, mix them and add HP20 macroporous resin for adsorption, elute with a gradient of 30%-75% ethanol, collect the eluent in segments, and obtain the polyene macrocyclic lactam compounds FW8-1 and FW8-4 in the 70-75% eluent.

[0022] Specifically, the method for preparing the polyene macrocyclic lactam compound by fermentation:

[0023] In step (3), the macroporous resin includes macroporous resin HP20;

[0024] In step (3), the mass ratio of the resin to the fermentation broth is 1:10-1:30;

[0025] In step (4), the mass ratio of the macroporous resin to the total amount of crude extract A and crude extract B is: 1:20～1: 40 .

[0026] Specifically, the method for preparing the polyene macrocyclic lactam compound by fermentation further includes a step of purifying the polyene macrocyclic lactam compound, specifically including:

[0027] (5) Refining: Take the polyene macrocyclic lactam compounds FW8-1 and FW8-4, and use C 18 Reversed-phase column chromatography was performed using a gradient elution with 40%-80% methanol-water solution to obtain FW8-1 and FW8-4 with purities greater than 80%; then, elution was performed with 30-40% acetonitrile-water solution to obtain FW8-1 and FW8-4 compounds with purities greater than 90%, respectively.

[0028] Preferably, in the refining step, C 18 Reversed-phase column chromatography includes Ailgent SB C18 The preparation column was eluted with 65% methanol-water at a flow rate of 5-10 ml / min and a detection wavelength of 290-300 nm to obtain the target substance with a purity greater than 80%. Then, it was eluted with 35% acetonitrile-water at a flow rate of 5-10 ml / min to finally obtain compounds FW8-1 and FW8-4 with a purity greater than 90%.

[0029] Specifically, in the method for preparing the polyene macrocyclic lactam compound by fermentation, step (4) and / or (5) further includes using Ailigent SB C18. The steps for HPLC monitoring of analytical columns;

[0030] Preferably, the solvent system is controlled as 30-40% acetonitrile-water, the analysis time is 50 min, the flow rate is 0.5-1.2 ml / min, and the detection wavelength is 290-300 nm.

[0031] This invention also discloses the use of the polyene macrocyclic lactam compound in the preparation of targeted drugs;

[0032] Preferably, the potential targets corresponding to the compound FW8-1 include: AKT2, ALK, AURKB, CDK2, CB1R, FGFR3, HDAC1 / HDAC2, JAK1, NAMPT or ZAP70;

[0033] Preferably, the potential targets of the compound FW8-4 include: AKT3, AURKB, BRD4, JAK1, NAMPT, PI3K-γ, or Renin.

[0034] This invention also discloses the potential use of the polyene macrocyclic lactam compound corresponding to the target in the preparation of antitumor, anti-inflammatory, anti-angiogenic, anti-fibrotic, blood pressure regulating, immunomodulatory, and neuroprotective drugs;

[0035] Preferably, the indications for compound FW8-1 include prostate cancer, breast cancer, ovarian cancer, non-small cell lung cancer, lymphoma, neuroblastoma, bladder cancer, multiple myeloma, T-cell lymphoma, lung cancer, rheumatoid arthritis, systemic lupus erythematosus, and inflammatory bowel disease.

[0036] Preferably, the indications for the compound FW8-4 include brain cancer, skin cancer, lung cancer, liver cancer, breast cancer, treatment of testicular nucleoprotein midline carcinoma, acute myeloid leukemia, rheumatoid arthritis, inflammatory bowel disease, as well as regulating blood pressure and protecting the cardiovascular system.

[0037] The polyene macrocyclic lactam compounds FW8-1 and FW8-4 described in this invention both have a polyene macrocyclic lactam structure, and due to an intramolecular addition reaction, they form a novel intramolecular bicyclic structure, providing a compound reserve for the research and development of new potential anti-tumor, anti-inflammatory, anti-angiogenic, anti-fibrotic, blood pressure regulating, immunomodulatory, and neuroprotective drugs.

[0038] The method for extracting and isolating polyene macrocyclic lactam compounds FW8-1 and FW8-4 from the fermentation broth of Micromonospora sp. FIM MA181224 is more suitable for large-scale production.

[0039] The polyene macrocyclic lactam compounds FW8-1 and FW8-4 described in this invention were subjected to virtual screening and biological activity prediction using the CODD-PRED database. The results showed that the compounds have potential antitumor, anti-inflammatory, and neuroprotective activities. This invention provides lead compounds for the research and development of new active drugs.

[0040] The polyene macrolides described in this invention, through virtual screening using the CODD-PRED database, revealed that FW8-1 possesses potential anticancer capabilities, including targets associated with prostate cancer, breast cancer, ovarian cancer, non-small cell lung cancer, lymphoma, neuroblastoma, bladder cancer, multiple myeloma, T-cell lymphoma, and lung cancer; as well as various autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, and inflammatory bowel disease. Virtual screening results indicate that FW8-4 exhibits target binding ability against brain cancers (such as malignant glioma) and skin cancers (such as melanoma), lung cancer, liver cancer, breast cancer, treatment of testicular nucleoprotein midline carcinoma, acute myeloid leukemia, rheumatoid arthritis, inflammatory bowel disease, and regulation of blood pressure and cardiovascular protection. Attached Figure Description

[0041] To make the content of this invention easier to understand, the invention will be further described in detail below with reference to specific embodiments and accompanying drawings, wherein...

[0042] Figure 1 This is the high-resolution mass spectrum of the macrocyclic lactam compound FW8-1 in this invention;

[0043] Figure 2 This is the proton nuclear magnetic resonance image of the macrocyclic lactam compound FW8-1 in this invention. 1 H spectrum;

[0044] Figure 3 This is the carbon NMR spectrum of the macrocyclic lactam compound FW8-1 in this invention. 13 C spectrum;

[0045] Figure 4 It is the macrocyclic lactam compound FW8-1 in this invention. 1 H- 1 HCOSY diagram;

[0046] Figure 5 This is the HSQC correlation spectrum of the macrocyclic lactam compound FW8-1 in this invention;

[0047] Figure 6 This is the HMBC correlation spectrum of the macrocyclic lactam compound FW8-1 in this invention;

[0048] Figure 7 This is the high-resolution mass spectrum of the macrocyclic lactam compound FW8-4 in this invention;

[0049] Figure 8 This is the proton NMR spectrum of the macrocyclic lactam compound FW8-4 in this invention. 1 H spectrum;

[0050] Figure 9 This is the carbon NMR spectrum of the macrocyclic lactam compound FW8-4 in this invention. 13 C spectrum;

[0051] Figure 10 It is the macrocyclic lactam compound FW8-4 in this invention. 1 H- 1 HCOSY diagram;

[0052] Figure 11 This is the HSQC correlation spectrum of the macrocyclic lactam compound FW8-4 in this invention;

[0053] Figure 12 This is the HMBC correlation spectrum of the macrocyclic lactam compound FW8-4 in this invention. Detailed Implementation

[0054] The marine micromonospora strain Micromonospora sp. FIM-MA181224 used in the following embodiments of the present invention is deposited at the China General Microbiological Culture Collection Center, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC NO.17139 and deposit date of January 7, 2019.

[0055] In the following embodiments of the present invention, novel polyene macrocyclic lactam compounds FW8-1 and FW8-4 were extracted and isolated from the fermentation broth of Micromonospora sp. FIM MA181224.

[0056] Example 1: Fermentation by Producing Microorganisms

[0057] Micromonospora sp. FIM MA181224 was inoculated onto starch-asparagine agar slant culture for activation and preservation, and then inoculated into liquid seed culture medium. After incubation at 30°C for 2 days, a shake-flask seed culture was obtained. The seed culture was inoculated into fermentation medium at a volume ratio of 10% and mixed, and cultured at 30°C with shaking for 5 days. The fermentation product was then collected.

[0058] The liquid seed culture medium comprises the following components in the indicated mass ratios: 1.8% soluble starch, 0.4% glucose, 0.4% peptone, 0.4% yeast extract, 0.03% MgSO4·7H2O, 0.08% NaCl, 0.03% (NH4)2SO4, 0.08% CaCO3, prepared with distilled water, and adjusted to pH 7.2.

[0059] The fermentation medium comprises the following components in the indicated mass ratios: 1.8% soluble starch, 0.5% glucose, 0.5% peptone, 0.2% soybean meal, 0.2% yeast powder, 0.05% K2HPO4·3H2O, 0.03% MgSO4·7H2O, 0.08% NaCl, 0.05% (NH4)2SO4, 0.1% CaCO3, 0.1% sea salt, prepared with tap water, and adjusted to pH 7.2.

[0060] Example 2: Extraction, purification, and refining of compounds FW8-1 and FW8-4

[0061] The fermentation products from Example 1 were collected for solid-liquid separation, and the fermentation broth and bacterial cells were collected separately.

[0062] Take the fermentation broth and add HP20 resin to mix. Control the volume ratio of the resin to the fermentation broth to be 1:20. After mixing, load the mixture onto a resin column for adsorption. After adsorption, wash with distilled water and then wash with 10-20% ethanol solution to remove impurities. Then perform full desorption with 75% ethanol (or perform gradient desorption with 30%-75% ethanol). After recovering the ethanol solvent, crude extract A is obtained.

[0063] The isolated mycelium was extracted three times with ethanol. The extract was collected and concentrated under reduced pressure to obtain crude extract B.

[0064] The crude extracts A and B were combined and dissolved in anhydrous ethanol. They were then adsorbed using HP20 macroporous resin and eluted with gradients of 30%, 40%, 50%, and 75% ethanol. Target substances FW8-1 and FW8-4 with a purity of approximately 50%-60% were obtained in the 50-75% elution zone.

[0065] Take the above compounds FW8-1 and FW8-4, and select Ailgent SB C18. The preparative column was used for purification and refining. Gradient elution with methanol-water (40%-80%) was performed at a flow rate of 3-8 ml / min. The target product was monitored at a wavelength of 290-300 nm and analyzed by analytical HPLC. Then, FW8-1 and FW8-4 with a purity greater than 80% were obtained in the 65% methanol-water elution zone.

[0066] Continue with the above compounds FW8-1 and FW8-4, using Ailgent SB C18 The column was purified by elution with 35% acetonitrile-water at a flow rate of 10 ml / min and a detection wavelength of 290-300 nm. Finally, compounds FW8-1 and FW8-4 with a purity greater than 90% were obtained. The collection time for compound FW8-1 was 14.5 min and the collection time for compound FW8-4 was 30.5 min.

[0067] In this embodiment, high-performance liquid chromatography (HPLC) can be used for detection and analysis during the extraction, purification, and refining processes. Specifically, Ailgent SB C18 is used. The analytical column was monitored by HPLC. The solvent system was 30-40% acetonitrile-water, the analysis time was 50 min, the flow rate was 1.0 ml / min, and the detection wavelength was 290-300 nm.

[0068] Structural analysis of compounds FW8-1 and FW8-4 in Example 3

[0069] In this embodiment, the structures of compounds FW8-1 and FW8-4 collected in Example 2 were identified and analyzed.

[0070] In this embodiment, the structure of the macrocyclic lactam compound FW8-1 was identified using MS and NMR techniques.

[0071] The physicochemical properties of compound FW8-1 are as follows: white amorphous solid, molecular formula: C 29 H 41 NO5, High-resolution mass spectrometry: Measured value: m / z 484.3063 [M+H] + The theoretical value is m / z 484.3058 [M+H]. + The compound FW8-1 is soluble in organic solvents such as methanol, acetone, acetonitrile, ethyl acetate, and dimethyl sulfoxide, and has an unsaturation degree of 10.

[0072] The compound FW8-1 was determined to have... 1 1H NMR spectrum (DMSO-d6, 600MHz): δ 7.49 (d, J = 8.9Hz, 1H), 6.82 (dd, J = 14.9, 11.4Hz, 1H), 6.47 (ddd, J = 15.0, 11.4, 1.9Hz, 1H), 6.36 (dd, J = 14.9, 10.8Hz, 1H), 6.14 (dd, J = 14.9, 11.3Hz, 1H), 6.09 (dd, J = 15.0, 10.9Hz, 1H), 5.89 (d, J = 15.5Hz, 1H), 5.81 (d, J = 11.4Hz, 1H), 5.75 (s, 1H), 5.72 (s, 1H), 5.56 (m, 2H), 5.50 (m ,1H),5.00(d,J=3.6Hz,1H),4.41(dq,J=10.3,5.6Hz,1H),4.30(d,J=4.6Hz, 1H),4.04(m,1H),3.92(m,2H),3.39(s,1H),2.96(q,J=9.6Hz,1H),2.09(td,J =9.6,6.5Hz,2H),1.90(dd,J=12.9,6.1Hz,1H),1.83(d,J=8.5Hz,6H),1.53(m ,2H),1.41(m,1H),1.23(s,1H),1.01(d,J=6.5Hz,3H),0.98(d,J=6.6Hz,3H).

[0073] The compound FW8-1 13C-NMR spectrum (DMSO-D6, 150MHz): δ163.86,141.63,138.48,138.15,134.76,134.54,134.30,133.84,133.68,132.63,130.16,130.00,128.77,126.02,124.48,85.80,76.21,75.77,71.80,71.76,45.47,44.35,41.91,38.92,28.73,23.23,17.96,17.01,14.57.

[0074] The compound FW8-1 1 H and 13 The C(DMSO-d6) attribution data are shown in Table 1 below.

[0075] Table 1. Compounds of FW8-1 1 H and 13 C(DMSO-d6) attribution

[0076]

[0077]

[0078] High-resolution mass spectrum and proton nuclear magnetic resonance spectrum of compound FW8-1 1 H spectrum, carbon nuclear magnetic resonance image 13 C-spectrum, HSQC related spectra 1 H- 1 The HCOSY spectrum and related HMBC spectra are attached. Figure 1-6 As shown.

[0079] The physicochemical properties of the compound FW8-4 are as follows: a pale yellow amorphous solid, with the molecular formula: C. 29 H 41 NO5, High-resolution mass spectrometry: Measured value: m / z 484.3050 [M+H] + The theoretical value is m / z 484.3058 [M+H]. + The compound FW8-4 is soluble in organic solvents such as methanol, acetone, acetonitrile, ethyl acetate, and dimethyl sulfoxide, and has an unsaturation degree of 10.

[0080] The compound FW8-4 16.51 H NMR spectrum (DMSO-d6, 600MHz): δ 6.92 (dd, J = 14.8, 11.3Hz, 1H), 6.51 (dd, J = 14.9, 10.8Hz, 1H), 6.31 (m, 1H), 6.26 (dd, J = 15.0, 11.3Hz, 1H), 6.23 (m, 1H), 6.20 (m, 1H), 6.16 (d, J = 14.8, 1H). 9Hz,1H),5.93(t,J=10.9Hz,1H),5.85(m,1H),5.66(dd,J=15.0,10.1Hz,1H),5.40–5.38 (m,3H),5.18(d,J=3.0Hz,1H),4.86(d,J=4.6Hz,1H),4.85(m,1H),4.63(d,J=7.2Hz,1H), 4.30(m,1H),4.20(m,1H),4.04(d,J=4.4Hz,1H),3.95(t,J=9.2Hz,1H),3.46(d,J=10.0H z,1H),2.08(td,J=9.7,5.7Hz,1H),2.02(ddd,J=13.5,8.9,4.7Hz,1H),1.78(s,3H),1.7 5(d,J=13.9Hz,1H),1.61(d,J=11.9Hz,1H),1.56(d,J=1.3Hz,3H),1.53(m,2H),1.32(d, J=6.4Hz, 3H), 1.19 (d, J=11.6Hz, 1H), 1.1 (dd, J=10.6, 3.9Hz, 1H), 1.02 (d, J=6.4Hz, 3H).

[0081] The compound FW8-4 13 C-NMR spectrum (DMSO-D6, 150MHz): δ165.05,140.81,139.38,138.02,136.36,136.23,134.70,134.04,133.55,130.84,130.21,130.18,125.93,125.28,123.61,77.09,74.84,68.68,66.47,63.45,53.10,46.65,46.56,40.06,37.58,21.01,18.06,17.84,14.55.

[0082] The compound FW8-4 1 H and 13 The C(DMSO-d6) attribution data are shown in Table 2 below.

[0083] Table 2. Compounds of FW8-4 1 H and13 C(DMSO-d6) attribution

[0084]

[0085]

[0086] High-resolution mass spectrum and proton nuclear magnetic resonance spectrum of compound FW8-4 1 H spectrum, carbon nuclear magnetic resonance image 13 C-spectrum, HSQC related spectra 1 H- 1 The HCOSY spectrum and related HMBC spectra are attached. Figure 7-12 As shown.

[0087] In summary, this embodiment determined the assignment of all carbon and hydrogen atoms in compounds FW8-1 and FW8-4, as well as the chemical structure of the compounds, confirming them as novel polyene macrocyclic lactam compounds. The specific structural formula is as follows:

[0088]

[0089] Example 4 Virtual screening and bioactivity prediction of compounds FW8-1 and FW8-4

[0090] In this embodiment, virtual target selection and biological activity prediction experiments were conducted on polyene macrocyclic lactam compounds FW8-1 and FW8-4.

[0091] The structures of the aforementioned polyene macrocyclic lactam compounds FW8-1 and FW8-4 were drawn and submitted to the CODD-PRED database for virtual screening to obtain the corresponding high-probability target types. Then, the biological activities of the corresponding targets were predicted, and the final output showed a theoretical value p Ic50 > 6.5. It can be seen that the potential targets corresponding to FW8-1 are: AKT2, ALK, AURKB, CDK2, CB1R, FGFR3, HDAC1 / HDAC2, JAK1, NAMPT, and ZAP70; the potential targets of compound FW8-4, as indicated by virtual screening, are: AKT3, AURKB, BRD4, JAK1, NAMPT, PI3K-γ, and Renin.

[0092] The predicted biological activities of the novel polyene macrocyclic lactam compounds FW8-1 and FW8-4 indicate that they possess potential biological activities against various tumor types, anti-autoimmune activity, and anti-inflammatory activity, thus providing lead compounds for the research and development of new polyene macrocyclic lactam compounds.

[0093] In summary, the polyene macrocyclic lactam compounds FW8-1 and FW8-4 of this invention both possess potential antitumor and anti-inflammatory functions. FW8-4, in particular, exhibits the ability to bind to potential targets for blood pressure regulation and neuroprotection. Therefore, FW8-1 and FW8-4 hold promise as antitumor and anti-inflammatory active substances.

[0094] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A polyene macrocyclic lactam compound, characterized in that, The polyene macrocyclic lactam compounds include compound FW8-1 as shown in formula (I) and / or compound FW8-4 as shown in formula (II):

2. A method for preparing the polyene macrocyclic lactam compound of claim 1 by fermentation, characterized in that, Includes the following steps: (1) Seed culture: Marine Micromonospora preserved on slant culture was inoculated into liquid seed culture medium and cultured at constant temperature. Seed culture was collected for later use. The marine micromonospora strain, classified as Micromonospora sp. FIM-MA181224, has been deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.17139. The liquid seed culture medium comprises the following components in the indicated mass percentages: soluble starch 1.5-2.0%, glucose 0.3-0.5%, peptone 0.3-0.5%, yeast extract 0.3-0.5%, MgSO4·7H2O 0.01-0.05%, NaCl 0.05-0.1%, (NH4)2SO4 0.01-0.05%, CaCO3 0.05-0.1%, pH 6.0-8.0; The temperature for the constant temperature culture step is 25-35℃, and the culture time is 1-3 days. (2) Fermentation broth culture: The seed liquid is transferred to the fermentation medium for constant temperature culture to obtain the fermentation product containing the desired polyene macrocyclic lactam compound. The fermentation medium comprises the following components in the indicated mass percentages: soluble starch 1.6-2.0%, glucose 0.3-0.6%, peptone 0.4-0.6%, soybean meal 0.2-0.3%, yeast powder 0.15-0.25%, K₂HPO₄·3H₂O 0.03-0.06%, MgSO₄·7H₂O 0.01-0.05%, NaCl 0.05-0.1%, (NH₄)₂SO₄ 0.03-0.06%, CaCO₃ 0.08-0.12%, and sea salt 0.8-0.12%. pH 6.2-8.0; The isothermal incubation step is performed at a temperature of 25-35℃, with shaking incubation for 3-6 days.

3. The method for preparing the polyene macrocyclic lactam compound by fermentation according to claim 2, characterized in that, The method further includes the steps of extraction and purification of the polyene macrocyclic lactam compound, specifically including: (3) Extraction: The collected fermentation products are separated into fermentation broth and mycelium respectively; the fermentation broth is added to macroporous resin for adsorption, and desorbed by ethanol solution to collect crude extract A; the mycelium is extracted by alcohol solvent, and the extract is collected and concentrated to obtain crude extract B; (4) Purification: Take crude extract A and crude extract B, mix them and add them to the macroporous resin for adsorption, elute with a gradient of 30%-75% ethanol, collect the eluent in segments, and obtain the polyene macrocyclic lactam compounds FW8-1 and FW8-4 in the 70-75% segment.

4. The method for preparing the polyene macrocyclic lactam compound by fermentation according to claim 3, characterized in that, The method further includes a step of purifying the polyene macrocyclic lactam compound, specifically including: (5) Purification: The polyene macrocyclic lactam compounds FW8-1 and FW8-4 were subjected to C18 reversed-phase column chromatography and gradient elution with 40%-80% methanol-water solution to obtain FW8-1 and FW8-4 with a purity greater than 80%; then eluted with 30-40% acetonitrile-water solution to obtain FW8-1 and FW8-4 compounds with a purity greater than 90%, respectively.

5. The method for preparing the polyene macrocyclic lactam compound by fermentation according to claim 4, characterized in that, Step (5) also includes using Ailgent SB C18 The steps for HPLC monitoring of analytical columns; The solvent system was controlled as 30-40% acetonitrile-water, the analysis time was 50 min, the flow rate was 0.5-1.2 ml / min, and the detection wavelength was 290-300 nm.

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