Neurospora parvosporium, salathromycin compound, and preparation method and application of neurospora parvosporium and salathromycin compound

Abstract

The invention discloses an acidophilic streptospora parvoides and salathromycin compound as well as a preparation method and application thereof, and belongs to the field of microbial products and medicine application. According to the invention, a group of salathromycin compounds are obtained from a fermentation product of the liquid fermentation of the acidosporium parvosporium (Catenulipora acidophilus), and the liquid fermentation is carried out on the acidosporium parvosporium (Catenulipora acidophilus) to obtain the salathromycin compounds. Experiments prove that the salathromycin compound provided by the invention has different degrees of inhibition effects on staphylococcus aureus and bacillus subtilis and has relatively strong inhibition activity on various tumor cells, and the half inhibitory concentration IC50 is a micromole level; it is indicated that the salathromycin compound is possibly further developed into a novel antibacterial and anti-tumor drug, it is indicated that the obtained salathromycin compound is expected to gain the application value in development of the antibacterial and anti-tumor drug, and the salathromycin compound has the potential of generating good social benefits and economic values.

Description

Technical Field

[0001] The present invention belongs to the field of microbial products and medical applications, and specifically relates to an acidophilic Neurospora parvum, a selamycin compound, and a preparation method and application thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance understanding of the overall background of the invention and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to those skilled in the art.

[0003] Chrolactomycins are a class of spirocyclic tetronic acid polyketides with a rare γ-pyrone skeleton that have attracted considerable attention due to their novel structure and promising biological activities. These compounds exhibit antibacterial activity and unique antitumor activity targeting telomerase.

[0004] Selamin compounds have the potential to become anti-tumor lead compounds targeting telomerase, but this type of natural product is rare. Currently, only three analogs, chrolactomycin, okilactomycin, and 6-hydroxychrolactomycin, have been reported, and the chemical diversity is seriously insufficient. Furthermore, the IC of existing selamin compounds (such as chrolactomycin) on solid tumor cells is still unclear. 50 It is generally greater than 10 μM, and the anti-tumor cytotoxic activity still needs to be improved.

[0005] Therefore, it is extremely necessary to discover and study new selamycin compounds in nature and apply them to the research and development of antibacterial and anti-tumor drugs. Summary of the Invention

[0006] In order to overcome the deficiencies of the prior art, the present invention aims to provide a Catenulispora acidiphila, a selamycin compound, and a preparation method and application thereof. The present invention isolates six selamycin compounds from the fermentation product of Catenulispora acidiphila, which have varying degrees of inhibitory effects on Staphylococcus aureus and Bacillus subtilis, and have strong inhibitory activity against a variety of tumor cells, with a half inhibitory concentration IC 50 The concentration of selamicin is at the micromolar level, indicating that the selamicin compounds provided by the present invention may be further developed into new antibacterial and antitumor drugs.

[0007] In order to achieve the above object, the technical solution of the present invention is:

[0008] The first aspect of the present invention provides a Catenulispora acidiphila, which was deposited in the General Microbiology Center of the China Culture Collection Administration on April 18, 2025, with a deposit number of CGMCC No.34268.

[0009] The second aspect of the present invention provides a use of the above-mentioned Catenulispora acidiphila in the production of selamycin compounds;

[0010] The structure of the seramycin compound is any one or more of the following:

[0011]

[0012] The third aspect of the present invention provides a seramycin compound, wherein the structure of the seramycin compound is any one or more of the following:

[0013]

[0014] A fourth aspect of the present invention provides a method for preparing the above-mentioned seramycin compounds, comprising:

[0015] fermenting Catenulispora acidiphila to obtain a fermentation product;

[0016] The fermentation product is separated and purified to obtain seramycin compounds.

[0017] In some embodiments of the invention, the fermentation comprises:

[0018] Catenulispora acidiphila is fermented in a culture medium at 25-35°C and 180-200 rpm. After 7-9 days, sterilized macroporous resin XAD-16 is added and culture is continued for 1-2 days to terminate the fermentation.

[0019] In some embodiments of the present invention, the culture medium is composed of 40-60 g / L soluble starch, 1-5 g / L yeast extract, 3-8 g / L peptone, and 5-15 g / L glucose.

[0020] In some embodiments of the present invention, the separation and purification comprises:

[0021] The fermentation product is centrifuged to collect the macroporous resin XAD-16 and the bacterial cells, methanol is added to resuspend the macroporous resin XAD-16 and the bacterial cells, and the extract is extracted and filtered to obtain an extract; the extract is spin-dried to obtain a crude extract;

[0022] The crude extract was dissolved in methanol and separated using a normal phase silica gel column with dichloromethane and methanol as solvents and gradient elution to obtain the fraction containing the target compound;

[0023] The components containing the target compound are purified by semi-preparative high performance liquid chromatography to obtain seramycin compounds.

[0024] In some embodiments of the present invention, the extraction comprises: adding methanol to resuspend the macroporous resin XAD-16 and the bacteria, extracting at 25-35° C. and 150-180 rpm for 1-5 hours, filtering to obtain an extract, repeating the extraction operation 2-5 times, and combining the extracts.

[0025] In some embodiments of the present invention, the conditions of the semi-preparative HPLC are as follows: the separation column model is Agilent ZORBAX SB-C18, 9.4×250 mm, 5 μm, the flow rate is 1.8-2.2 mL / min, phase A is ultrapure water, and phase B is acetonitrile; the elution program is: 0-4 min, 70% B; 4-30 min, 70%-100% B; 30-40 min, 100% B or 0-4 min, 70% B; 4-25 min, 70%-94% B; 25-25.1 min, 94%-100% B; 25.1-36 min, 100% B.

[0026] A fifth aspect of the present invention provides use of the above-mentioned seramycin compounds in the preparation of antibacterial and / or antitumor drugs.

[0027] In some embodiments of the present invention, the bacteria include Staphylococcus aureus and Bacillus subtilis.

[0028] In some embodiments of the present invention, the tumor comprises at least one of renal clear cell adenocarcinoma, esophageal cancer, bladder cancer, thyroid cancer, breast cancer, osteosarcoma, and malignant melanoma.

[0029] The beneficial effects of the present invention are:

[0030] The present invention separates a strain of Catenulispora acidiphila capable of producing selamycin compounds from a symbiotic bacterium of a moss. After liquid fermentation, the fermentation product of the strain contains selamycin compounds with a new structure, thereby expanding the types of natural selamycin compounds.

[0031] It has been verified that the selamycin compounds provided by the present invention have different degrees of inhibitory effects on Staphylococcus aureus and Bacillus subtilis, and have strong inhibitory activity against various tumor cells, with a half inhibitory concentration IC 50The concentration of seramycin in the micromolar range indicates that the seramycin compounds may be further developed into new antibacterial and antitumor drugs, and that the seramycin compounds obtained in the present invention are expected to increase their application value in the development of antibacterial and antitumor drugs, and have the potential to generate good social benefits and economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The accompanying drawings, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0033] Figure 1 is the 16S rDNA phylogenetic tree of Catenulispora acidiphila TX15 in Example 1 of the present invention;

[0034] Figure 2 The liquid fermentation results of Catenulispora acidiphila TX15 in Example 2 of the present invention; wherein 1 corresponds to selamycin A, 2 corresponds to selamycin B, 3 corresponds to selamycin C, 4 corresponds to selamycin D, 5 corresponds to selamycin E, and 6 corresponds to selamycin F;

[0035] Figure 3 The structures of seramycin compounds AF obtained in Example 2 of the present invention;

[0036] Figure 4 These are the key COSY and HMBC correlation signals of seramycin-based compounds AF obtained in Example 2 of the present invention;

[0037] Figure 5 is the NOESY correlation signal of the seramycin-based compounds AF obtained in Example 2 of the present invention;

[0038] Figure 6 This is a high-resolution mass spectrum of seramycin compounds AC obtained in Example 2 of the present invention;

[0039] Figure 7 This is a high-resolution mass spectrum of the seramycin compound DF obtained in Example 2 of the present invention. DETAILED DESCRIPTION

[0040] Biological Deposit Description

[0041] Culture name: Catenulispora acidiphila, deposited on April 18, 2025 in the General Microbiology Center of China Culture Collection Administration, the address of the collection center is: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences; the collection number is CGMCC No. 34268.

[0042] The TX15 mentioned in this application is the strain with the deposit number CGMCC No.34268.

[0043] In view of the fact that there are few types of existing natural selamin compounds, the antibacterial and antitumor activities still need to be improved, and the current situation that existing antibacterial and antitumor drugs are in urgent need of development, the present invention discloses acidophilic Neurospora microspora, selamin compounds and their preparation methods and applications. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art and they are all considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0044] The present invention isolated a strain of Catenulispora acidiphila TX15 from a moss symbiotic bacterium that can produce selamin-like compounds. The identification results are as follows:

[0045] 1. Morphological identification

[0046] Gram-positive bacteria, colonies are round in shape and white in color.

[0047] 2. Molecular Identification

[0048] The 16S sequence of this strain is:

[0049] CCGCCTTCGGGCGGGGATCAGTGGCGAACGGGTGAGTAACACGTGGGTAACCTG

[0050] CCCTCCACTCTGGGATAACTCCGGGAAACCGGGGCTAATACCGGATACGACCTGA

[0051] GGCGGCATCGCCTCGGGTGGAAAGTTTTTCGGTGGAGGATGGACCCGCGGCCTAT

[0052] CAGCTTGTTGGTGGGGTAAAGGCCTACCAAGGCGATGACGGGTAGCCGGCCTGA

[0053] GAGGGCGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGG

[0054] CAGCAGTGGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCGACGCCGCGT

[0055] GAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAGCAGGGACGAAGCGCAAG

[0056] TGACTGTACCTGCAGAAGAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAAT

[0057] ACGTAGGGTGCGAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCTTGTAGGTGGC

[0058] TCGTTGCGTCGGGAGTGAAAACCCGCGGCTTAACCGCGGGCCTGCTTCCGATACG

[0059] GGCGAGCTAGAGTTCGGCAGGGGAGACTGGAATTCCTGGTGTAGCGGTGGAATG

[0060] CGCAGATATCAGGAGGAACACCGGTGGCGAAGGCGGGTCTCTGGGCCGATACTG

[0061] ACACTGAGAAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCC

[0062] ACGCCGTAAACGTTGGGAACTAGGTGTGGGGTCCATTCCACGGGCTCTGTGCCGC

[0063] AGCTAACGCATTAAGTTCCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCA

[0064] AAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGTGGCTTAATTCGATGC

[0065] AACGCGAAGAACCTTACCTGGGCTTGACATGTTCGGTCTACCTGCAGAGATGTGG

[0066] GGTGCTTTTGCGCCGTTCACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGA

[0067] GATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGTTCCATGTTGCCAGCGCG

[0068] TTATGGCGGGGACTCATGGGAGACTGCCGGGGTCAACTCGGAGGAAGGTGGGGA

[0069] TGACGTCAAATCATCATGCCCCTTATGTCCAGGGCTGCACACATGCTACAATGGCC

[0070] GGTACAGAGGGCTGCGATACCGCAAGGTGGAGCGAATCCCAAAAAGCCGGTCTC

[0071] AGTTCGGATCGGGGTCTGCAACTCGACCCCGTGAAGTCGGAGTCGCTAGTAATCG

[0072] CAGATCAGCAACGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACGTCACGAAAGTCGGTAACACCCGAAGCCGGTGGCCT(SEQ ID NO.1)

[0073] Based on its 16S rDNA sequence, a phylogenetic tree was constructed as Figure 1 shown, indicating that this strain has the closest genetic relationship with Catenulispora acidiphila strain DSM 44928.

[0074] Morphological characteristics and homology sequence analysis indicated that the strain belongs to the kingdom Bacteria, the genus Catenulispora, and the species acidiphila. It was deposited at the General Microbiology Center of the China Culture Collection Administration (CGMCC), Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, on April 18, 2025, under the accession number CGMCC No. 34268, and named Catenulispora acidiphila TX15.

[0075] The present invention isolated and identified a group of novel chrolactomycin compounds (named chrolactomycin A, chrolactomycin B, chrolactomycin C, chrolactomycin D, chrolactomycin E, and chrolactomycin F) from the fermentation extract of a rare actinomycete, Catenulispora acidiphila TX15. Selamycin D, chrolactomycin E, and chrolactomycin F are novel dimeric backbone compounds, and their cytotoxic activity against various tumor cells is superior to that of chrolactomycin.

[0076] Therefore, a second typical embodiment of the present invention provides the use of the above-mentioned Catenulispora acidiphila in the production of selamycin compounds;

[0077] The structure of the seramycin compound is any one or more of the following:

[0078]

[0079] A third typical embodiment of the present invention provides a seramycin compound, wherein the structure of the seramycin compound is any one or more of the following:

[0080]

[0081]

[0082] The present invention has demonstrated through experiments that selamycin AC has different degrees of inhibitory effects on Staphylococcus aureus ATCC 29213 and Bacillus subtilis ATCC 6633. Selamycin AF has better cytotoxic activity against various tumor cells than chrolactomycin. Among them, selamycin CE has good cytotoxic activity against human renal clear cell adenocarcinoma cells (786-O), human esophageal cancer cells (TE-1), human bladder cancer cells (5637), human thyroid cancer cells (CAL-62), human breast cancer cells (MCF7), human osteosarcoma cells (HOS) and human malignant melanoma cells (A-375), with a half inhibitory concentration IC50 Selamicin B has good cytotoxic activity against human renal clear cell adenocarcinoma cells (786-O), human esophageal cancer cells (TE-1), human bladder cancer cells (5637), human breast cancer cells (MCF7) and human osteosarcoma cells (HOS), with a half inhibitory concentration IC 50 Selamicin F has cytotoxic activity against human renal clear cell adenocarcinoma cells (786-O), human esophageal cancer cells (TE-1), human bladder cancer cells (5637) and human thyroid cancer cells (CAL-62), with a half inhibitory concentration IC 50 It is at the micromolar level. It can be seen that the selamycin compounds obtained by the present invention may be further developed into new antibacterial and antitumor drugs, indicating that the selamycin compounds obtained by the present invention are expected to increase their application value in the development of antibacterial and antitumor drugs and have the potential to generate good social benefits and economic value.

[0083] It should be noted that the present invention further provides a structural formula of selamicycin AF using Arabic numerals to position carbon atoms in the chemical structure, as shown below:

[0084]

[0085] The selamycin compounds are obtained from the fermentation products of Catenulispora acidiphila through liquid fermentation.

[0086] Therefore, a fourth typical embodiment of the present invention provides a method for preparing the above-mentioned seramycin compounds, comprising:

[0087] fermenting Catenulispora acidiphila to obtain a fermentation product;

[0088] The fermentation product is separated and purified to obtain seramycin compounds.

[0089] In some examples of this embodiment, the fermenting comprises:

[0090] Catenulispora acidiphila is fermented in a culture medium at 25-35°C and 180-200 rpm. After 7-9 days, sterilized macroporous resin XAD-16 is added and culture is continued for 1-2 days to terminate the fermentation.

[0091] In some examples of this embodiment, the culture medium is composed of 40-60 g / L soluble starch, 1-5 g / L yeast extract, 3-8 g / L peptone, and 5-15 g / L glucose.

[0092] In order to improve the fermentation efficiency, the formula of the culture medium is preferably: 50 g / L soluble starch, 3 g / L yeast extract, 5 g / L peptone, and 10 g / L glucose.

[0093] It should be noted that the term soluble starch, also known as soluble starch in English, is a starch derivative obtained by treating starch with oxidants, acids, glycerol, enzymes or other methods. Its chemical formula is (C6H 10 O5) n , CAS registration number is 9005-84-9, EINECS registration number is 232-686-4, it is a white or off-white powder, odorless and tasteless, a conventional commercial product, and can be purchased.

[0094] It should be noted that yeast extract, also known as yeast flavor, is abbreviated as YE in English. It is a pure natural product that comes in a brownish-yellow soluble paste or light yellow powder form, and is made from protein-rich edible yeast using modern biotechnology, such as autolysis, enzymatic hydrolysis, separation, and concentration, to degrade and refine the proteins and nucleic acids within yeast cells. It is commonly used in food, cosmetics, and health supplements. It is a commercially available product.

[0095] It should be noted that the full name of the macroporous resin XAD-16 is non-ionic macroporous resin XAD16, CAS number: 9003-69-4, molecular formula: C 10 H 10 , molecular weight: 130.19, MDL number: MFCD00145831. It is a conventional commercial product and can be purchased.

[0096] In some examples of this embodiment, the separation and purification comprises:

[0097] The fermentation product is centrifuged to collect the macroporous resin XAD-16 and the bacterial cells, methanol is added to resuspend the macroporous resin XAD-16 and the bacterial cells, and the extract is extracted and filtered to obtain an extract; the extract is spin-dried to obtain a crude extract;

[0098] The crude extract was dissolved in methanol and separated using a normal phase silica gel column with dichloromethane and methanol as solvents and gradient elution to obtain the fraction containing the target compound;

[0099] The components containing the target compound are purified by semi-preparative high performance liquid chromatography to obtain seramycin compounds.

[0100] In some examples of this embodiment, the extraction includes: adding methanol to resuspend the macroporous resin XAD-16 and the bacteria, extracting at 25-35°C and 150-180 rpm for 1-5 hours, filtering to obtain an extract, repeating the extraction operation 2-5 times, and combining the extracts.

[0101] In some examples of this embodiment, the conditions of the semi-preparative high performance liquid chromatography are as follows: the separation column model is Agilent ZORBAX SB-C18, 9.4×250 mm, 5 μm, the flow rate is 1.8-2.2 mL / min, phase A is ultrapure water, and phase B is acetonitrile; the elution program is: 0-4 min, 70% B; 4-30 min, 70%-100% B; 30-40 min, 100% B or 0-4 min, 70% B; 4-25 min, 70%-94% B; 25-25.1 min, 94%-100% B; 25.1-36 min, 100% B.

[0102] In some examples of this embodiment, the preparation method comprises the following steps:

[0103] Catenulispora acidiphila was inoculated onto a MS solid plate at 30°C. After the bacteria grew, they were scraped and cultured in YEME liquid and cultured overnight at 30°C to obtain a fermentation seed solution.

[0104] The fermentation seed liquid was transferred to YEME fermentation medium at an inoculum rate of 2%, v / v, at 30°C, 200 rpm, and after 8 days, sterilized macroporous resin XAD-16 was added. The culture was continued for 1 day, and the fermentation was completed to obtain the fermentation product.

[0105] The fermentation product was centrifuged to collect the macroporous resin XAD-16 and the bacterial cells, and then methanol was added to resuspend the macroporous resin XAD-16 and the bacterial cells. The mixture was extracted at 30°C and 180 rpm for 3 hours, and the extract was filtered to obtain the extract. The extraction operation was repeated three times. The extracts were combined and rotary evaporated to dryness to obtain a crude extract.

[0106] The crude extract was dissolved in methanol and initially separated using a normal phase silica gel column with dichloromethane and methanol as solvents by gradient elution to obtain two fractions containing the target compound, named NP-1 and NP-2.

[0107] NP-1 and NP-2 were purified by semi-preparative HPLC. The separation column model equipped with the instrument was Agilent ZORBAX SB-C18, 9.4×250 mm, 5 μm, the flow rate was 2 mL / min, phase A was ultrapure water (Milli-Q H2O), and phase B was acetonitrile; the separation procedure of NP-1 was: 0-4 min, 70% B; 4-30 min, 70%-100% B; 30-40 min, 100% B; the separation procedure of NP-2 was: 0-4 min, 70% B; 4-25 min, 70%-94% B; 25-25.1 min, 94%-100% B; 25.1-36 min, 100% B; to obtain seramycin compounds.

[0108] The formula of MS medium is: D-mannitol 20g / L, cooked soybean powder 20g / L, agar powder 20g / L. The formula of YEME medium is: soluble starch 50g / L, yeast extract 3g / L, peptone 5g / L, glucose 10g / L.

[0109] A fifth typical embodiment of the present invention provides the use of the above-mentioned seramycin compounds in the preparation of antibacterial and / or antitumor drugs.

[0110] In some examples of this embodiment, the bacteria include but are not limited to Staphylococcus aureus and Bacillus subtilis.

[0111] In some examples of this embodiment, the tumor includes but is not limited to at least one of renal clear cell adenocarcinoma, esophageal cancer, bladder cancer, thyroid cancer, breast cancer, osteosarcoma, and malignant melanoma.

[0112] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0113] If no specific conditions are specified in the examples, the experiments were carried out under conventional conditions or those recommended by the manufacturer. Other experimental methods, reagents, or instruments, unless otherwise specified, are conventional methods in the relevant fields and commercially available conventional products.

[0114] Example 1: Liquid fermentation of Catenulispora acidiphila TX15

[0115] Acidophilic Neurospora tenuissima TX15 was inoculated onto a MS solid plate at 30°C. After bacterial growth, the cells were scraped and cultured in YEME liquid (50mL / 250mL) and incubated at 30°C overnight to obtain a fermentation seed solution. The fermentation seed solution was transferred to a 250mL Erlenmeyer flask containing 50mL of YEME fermentation medium at a 2% (v / v) inoculum. A total of 30L was fermented at 30°C, 200rpm, and sterilized macroporous resin XAD-16 was added after 8 days. The culture was continued for 1 day, and the fermentation product was obtained and the compound was extracted.

[0116] The fermentation product was centrifuged at 8000 rpm for 5 minutes to collect the macroporous resin XAD-16 and the bacterial cells. 1 L of methanol was then added to resuspend the macroporous resin XAD-16 and the bacterial cells. Extraction was performed at 30°C and 180 rpm for 3 hours. The extract was filtered to obtain the extract. The extraction process was repeated three times. The extracts were combined and rotary evaporated to dryness to obtain a crude extract.

[0117] The above-mentioned MS culture medium formula is: D-mannitol 20g / L, cooked soybean powder 20g / L, and agar powder 20g / L.

[0118] The YEME culture medium formula is: 50 g / L soluble starch, 3 g / L yeast extract, 5 g / L peptone, and 10 g / L glucose.

[0119] Example 2: Isolation, purification and structural identification of seramycin-like compounds

[0120] 2.1 Isolation and purification of seramycin-like compounds

[0121] The crude extract obtained in Example 1 was dissolved in a small amount of methanol and initially separated using a normal phase silica gel column with dichloromethane and methanol as solvents by gradient elution to obtain two fractions containing the target compound, which were named NP-1 and NP-2.

[0122] NP-1 and NP-2 were purified by semi-preparative HPLC using an Agilent ZORBAX SB-C18 column (9.4 × 250 mm, 5 μm) at a flow rate of 2 mL / min. Phase A consisted of Milli-Q HO and phase B consisted of acetonitrile. The separation profile for NP-1 was: 0-4 min, 70% B; 4-30 min, 70%-100% B; 30-40 min, 100% B. The separation profile for NP-2 was: 0-4 min, 70% B; 4-25 min, 70%-94% B; 25-25.1 min, 94%-100% B; 25.1-36 min, 100% B. Six compounds were purified and designated seramycin AF. The yields of these six compounds are shown in Table 1.

[0123] Table 1 Yield of Selamicin AF

[0124] Selamicin Yield Selamicin Yield A 7.7mg D 9.4mg B 6.6mg E 4.0mg C 3.0mg F 3.0mg

[0125] 2.2 Structural identification of selamycin-like compounds

[0126] The selamin-like compounds separated in 2.1 were further tested by high-resolution mass spectrometry, nuclear magnetic resonance, and optical rotation.

[0127] Instrument models: Bruker Impact HD microTOF Q III mass spectrometer was used for high-resolution mass spectrometry; Bruker Avance NEO 600 MHz was used for nuclear magnetic resonance measurements; and Rudolph AUTOPOL III fully automatic polarimeter was used for optical rotation measurements.

[0128] The physicochemical properties and spectral data of the seramycin compounds obtained in this example are as follows:

[0129] Seramycin A: white powder; [α] 20 D -27(c 0.1,MeOH);UV(MeOH):λ max 217nm; HRMS(ESI)m / z:451.2327[M+H] + (Calculated value C 24 H 35 O8,451.2326); 1 H and 13 C-NMR data are shown in Table 2.

[0130] Seramycin B: white powder; [α] 20 D -18(c 0.1,MeOH);UV(MeOH):λ max 221nm; HRMS(ESI)m / z:465.2483[M+H] + (Calculated value C 25 H 37 O8,465.2483); 1 H and 13 C-NMR data are shown in Table 2.

[0131] Seramycin C: white powder; [α] 20 D -18(c 0.1,MeOH);UV(MeOH):λ max 221nm,352nm; HRMS(ESI)m / z:570.2693[M+H] + (Calculated value C 31 H 40 NO9,570.2698);1 H and 13 C-NMR data are shown in Table 3.

[0132] Seramycin D: white powder; [α] 20 D +6(c 0.1,MeOH);UV(MeOH):λ max 212nm; HRMS(ESI)m / z:993.4604[M+H] + (Calculated value C 52 H 69 N2O 17 ,993.4591); 1 H and 13 C-NMR data are shown in Table 3.

[0133] Seramycin E: white powder; [α] 20 D -8(c 0.1,MeOH);UV(MeOH):λ max 220nm; HRMS(ESI)m / z:993.4590[M+H] + (Calculated value C 52 H 69 N2O 17 993.4591); 1 H and 13 C-NMR data are shown in Table 4.

[0134] Seramycin F: white powder; [α] 20 D +4(c 0.1,MeOH);UV(MeOH):λ max 224nm; HRMS(ESI)m / z:979.4412[M+H] + (Calculated value C 51 H 67 N2O 17 979.4434); 1 H and 13 C-NMR data are shown in Table 5.

[0135] Table 2 Selamycin A and in DMSO-d6 1 H (600MHz) and 13 C (150 MHz) NMR data

[0136]

[0137]

[0138] a Indicates overlapping signals.

[0139] Table 3 Seramycin C and Seramycin D in DMSO-d6 1 H (600MHz) and 13 C (150 MHz) NMR data

[0140]

[0141]

[0142]

[0143] a Indicates overlapping signals.

[0144] Table 4 Selamicin E in DMSO-d6 1 H (600MHz) and 13 C (150 MHz) NMR data

[0145]

[0146]

[0147] a Indicates overlapping signals.

[0148] Table 5 Selamycin F in DMSO-d6 1 H (600MHz) and 13 C (150 MHz) NMR data

[0149]

[0150]

[0151] a Indicates overlapping signals.

[0152] The molecular formula of seramycin A was estimated to be C based on high resolution mass spectrometry (HRESIMS). 24 H 34 O8. Seramycin A 1 H and 13 The C NMR data (Table 2) gave the corresponding 24 carbon signals, including one keto carbonyl (δ C 203.0), 2 ester carbonyl groups (δ C 168.2, 167.9), 2 olefinic carbons (δ H / C 6.57 / 138.0, 133.6), two oxygen-linked quaternary carbons (δ c 83.5, 79.5), 7 methines (δ H / C4.03 / 84.0, 4.54 / 83.1, 2.86 / 52.7, 2.39 / 41.4, 1.99 / 33.0, 1.82 / 29.7, 2.39 / 26.9), 6 methylene groups (δ H / C 3.94,3.81 / 57.5,1.38,0.86 / 44.6,1.12,0.97 / 37.2,1.67,1.23 / 33.6,1.49,1.21 / 31.7,1.55 / 23.4), 4 methyl groups (δ H / C 3.53 / 54.6, 0.84 / 23.8, 1.00 / 19.9, 0.89 / 18.7). Further through 1 H- 1 H COSY and HMBC related signals ( Figure 4 ) determined the planar structure of seramycin A, which is a novel seramycin analog. Considering that seramycin A and the known compound chrolactomycin have the same biosynthetic pathway, the stereo configuration of the shared chiral center should be consistent with that of the known compound chrolactomycin, and combined with the NOESY related signals ( Figure 5 ) were determined to be 1S, 5R, 7R, 8R, 9S, 11S, 12S, 13S, and 15S. The structures of seramycin B and seramycin C were deduced in the same way, which are methoxy and anthranilic acid substituted derivatives of seramycin A, respectively.

[0153] The molecular formula of selamycin D was estimated to be C based on high resolution mass spectrometry (HRESIMS). 52 H 69 N2O 17 ,and 13 The C NMR spectrum showed only 27 carbon signals (Table 3), suggesting that seramycin D may be a symmetrical dimer structure. 1 H- 1 H COSY and HMBC related signals ( Figure 4 ) determined the planar structure of seramycin D, which is a dimeric seramycin compound with a new skeleton formed by the polymerization of one molecule of barbituric acid and two molecules of seramycin monomers. Considering that seramycin D and seramycin A have the same NOESY related signals ( Figure 5 ), thus determining that the absolute configuration of seramycin D is consistent with that of seramycin A. 1 H and 13 C NMR data (Table 4) and 1 H- 1 H COSY and HMBC related signals ( Figure 4) showed that seramycin E and seramycin D had the same planar structure, but the chemical shift of seramycin E was significantly different from that of seramycin D, suggesting that seramycin E and seramycin D had different stereo configurations. Further analysis of NOESY related signals ( Figure 5 ) It was determined that C-9′ in seramycin E was in R configuration, which was opposite to 9′S in seramycin D, while the stereo configurations of other chiral centers were the same.

[0154] 1 H and 13 C NMR data (Table 5) and 1 H- 1 H COSY and HMBC related signals ( Figure 4 ) showed that seramycin F also has a dimeric structure, and the main difference from seramycin D is the lack of the 21′ methyl group. NOESY related signals ( Figure 5 ) confirmed that seramycin F and seramycin D have the same stereo configuration.

[0155] Example 3: Antibacterial and anti-tumor cytotoxic activity testing of seramycin compounds

[0156] 3.1 Antimicrobial activity test

[0157] The test sample was seramycin AF isolated and purified in Example 2. An appropriate amount of sample was accurately weighed and prepared into a 10 mg / mL solution in DMSO. The antibacterial activity was determined using the two-fold dilution method. The antibacterial activity results are shown in Table 6.

[0158] Table 6 Antibacterial activity (MIC / μg / mL)

[0159]

[0160] As shown in Table 6, seramycin AC has a certain inhibitory effect on Staphylococcus aureus ATCC 29213 and Bacillus subtilis ATCC6633.

[0161] Comparative experiments showed that selamycin B had the best antibacterial activity, outperforming the other compounds. Since selamycin AC exhibited antibacterial activity, it was shown that the selamycin compounds had the potential to be developed into antibacterial drugs.

[0162] 3.2 Antitumor cytotoxic activity test

[0163] The test sample was seramycin AF isolated and purified in Example 2. An appropriate amount of sample was accurately weighed and prepared with DMSO to a solution of the desired concentration. The anti-tumor cytotoxic activity was determined using the MTT assay. The anti-tumor cytotoxic activity results are shown in Table 7.

[0164] Table 7 Antitumor cytotoxic activity (IC50 / μM)

[0165]

[0166]

[0167] As shown in Table 7, selamycin CE has good cytotoxic activity against human renal clear cell adenocarcinoma cells (786-O), human esophageal cancer cells (TE-1), human bladder cancer cells (5637), human thyroid cancer cells (CAL-62), human breast cancer cells (MCF7), human osteosarcoma cells (HOS) and human malignant melanoma cells (A-375), with a half inhibitory concentration IC 50 Selamicin B has good cytotoxic activity against human renal clear cell adenocarcinoma cells (786-O), human esophageal cancer cells (TE-1), human bladder cancer cells (5637), human breast cancer cells (MCF7) and human osteosarcoma cells (HOS), with a half inhibitory concentration IC 50 Selamicin F has cytotoxic activity against human renal clear cell adenocarcinoma cells (786-O), human esophageal cancer cells (TE-1), human bladder cancer cells (5637) and human thyroid cancer cells (CAL-62), with a half inhibitory concentration IC 50 At the micromolar level.

[0168] Comparative experiments showed that the dimeric compound chrolactomycin D exhibited the best antitumor cytotoxic activity, surpassing all monomeric compounds. Furthermore, chrolactomycin D and chrolactomycin E exhibited higher antitumor activity than the known compound chrolactomycin. The highly potent antitumor cytotoxic activity of chrolactomycin D and chrolactomycin E suggests that these chrolactomycin compounds have the potential to be developed into novel antitumor drugs.

[0169] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. Catenulispora acidiphila, characterized in that It was deposited in the General Microbiology Center of China Culture Collection Administration on April 18, 2025, with the deposit number CGMCC No.34268.

2. Use of the Catenulispora acidiphila according to claim 1 in the production of selamycin-related compounds; The structure of the seramycin compound is any one or more of the following:

3. A seramycin compound, characterized in that: The structure of the seramycin compound is any one or more of the following:

4. The method for preparing the seramycin compound according to claim 3, characterized in that: include: fermenting Catenulispora acidiphila to obtain a fermentation product; The fermentation product is separated and purified to obtain seramycin compounds.

5. The preparation method according to claim 4, wherein The fermentation comprises: Catenulispora acidiphila is fermented in a culture medium at 25-35°C and 180-200 rpm. After 7-9 days, sterilized macroporous resin XAD-16 is added and culture is continued for 1-2 days to terminate the fermentation.

6. The preparation method according to claim 5, wherein The culture medium comprises 40-60 g / L of soluble starch, 1-5 g / L of yeast extract, 3-8 g / L of peptone, and 5-15 g / L of glucose.

7. The preparation method according to claim 4, wherein The separation and purification comprises: The fermentation product is centrifuged to collect the macroporous resin XAD-16 and the bacterial cells, methanol is added to resuspend the macroporous resin XAD-16 and the bacterial cells, and the extract is extracted and filtered to obtain an extract; the extract is spin-dried to obtain a crude extract; The crude extract was dissolved in methanol and separated using a normal phase silica gel column with dichloromethane and methanol as solvents and gradient elution to obtain the fraction containing the target compound; The components containing the target compound are purified by semi-preparative high performance liquid chromatography to obtain seramycin compounds.

8. The preparation method according to claim 7, wherein The extraction comprises: adding methanol to resuspend the macroporous resin XAD-16 and the bacteria, extracting at 25-35° C. and 150-180 rpm for 1-5 hours, filtering to obtain an extract, repeating the extraction operation 2-5 times, and combining the extracts.

9. The preparation method according to claim 7, wherein The conditions for the semi-preparative high performance liquid chromatography are as follows: the separation column model is Agilent ZORBAX SB-C18, 9.4×250 mm, 5 μm, the flow rate is 1.8-2.2 mL / min, phase A is ultrapure water, and phase B is acetonitrile; the elution program is: 0-4 min, 70% B; 4-30 min, 70%-100% B; 30-40 min, 100% B or 0-4 min, 70% B; 4-25 min, 70%-94% B; 25-25.1 min, 94%-100% B; 25.1-36 min, 100% B.

10. Use of the seramycin compound according to claim 3 in the preparation of antibacterial and / or antitumor drugs; Preferably, the bacteria include Staphylococcus aureus and Bacillus subtilis; Preferably, the tumor includes at least one of renal clear cell adenocarcinoma, esophageal cancer, bladder cancer, thyroid cancer, breast cancer, osteosarcoma and malignant melanoma.

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