CN117447471A - Indole diketopiperazine compound, preparation method thereof and application thereof in preparation of osteoclast differentiation inhibitor - Google Patents

Indole diketopiperazine compound, preparation method thereof and application thereof in preparation of osteoclast differentiation inhibitor Download PDF

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CN117447471A
CN117447471A CN202311457989.4A CN202311457989A CN117447471A CN 117447471 A CN117447471 A CN 117447471A CN 202311457989 A CN202311457989 A CN 202311457989A CN 117447471 A CN117447471 A CN 117447471A
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preparation
indole
diketopiperazine compound
indole diketopiperazine
prenylcyclotryprostatin
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刘昕明
罗小卫
林妙萍
谭艳辉
彭帅
王佳敏
高程海
刘永宏
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Guangxi University of Chinese Medicine
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Abstract

The invention provides an indole diketopiperazine compound, a preparation method thereof and application thereof in preparing an osteoclast differentiation inhibitor, and discloses an indole diketopiperazine compound named as presbycylotrypostatin B, which has a structural formula shown as a formula (I), has a remarkable inhibition effect on LPS-induced NF- κB luciferase at a concentration of 10 mu M, can remarkably inhibit RANKL-induced osteoclast precursor cells BMMs from differentiating into osteoclasts, has no obvious cytotoxicity, and is an ideal candidate compound developed into a novel osteoclast differentiation inhibitor drug.

Description

Indole diketopiperazine compound, preparation method thereof and application thereof in preparation of osteoclast differentiation inhibitor
Technical Field
The invention relates to the field of marine medicaments, in particular to an indole diketopiperazine compound, a preparation method thereof and application thereof in preparation of an osteoclast differentiation inhibitor.
Background
Osteoporosis is a common degenerative disease of the bone, which is mainly manifested by increased bone fragility, reduced bone mass and destruction of bone microstructure, severely threatening the health of the aging population and postmenopausal women. Osteoclasts (OCs) are special cells formed by the fusion of monocyte/macrophage hematopoietic lineage precursor cells, the only cells in humans that have bone resorption function. The defect of osteoclast activity leads to bone sclerosis and bone marrow failure, and excessive activation can lead to osteolytic diseases such as osteoporosis, rheumatoid arthritis, tumor bone metastasis and the like, and inhibition of the formation and resorption functions of OCs is one of the main strategies for treating osteoporosis.
The formation of osteoclasts is a stepwise process initiated by the binding of a receptor activator of the nuclear factor- κb receptor activator ligand (Receptor activator of nuclear kappa B ligand, RANKL) to its receptor RANK on the monocyte/macrophage precursor. The related inhibitors of osteoclast differentiation applied clinically at present are mainly denoxib and bisphosphonate drugs, but have certain complications and side effects. There are few studies on marine-derived anti-osteoclast differentiation inhibitors, and only some compounds have been publicly reported to have an anti-osteoclast differentiation effect, as in chinese patent application 1: CN201610478740.5, "application of marine-source nitrophenyl ester sesquiterpenoids in preparation of osteoclast differentiation inhibitors"; chinese patent application 2: CN202310241947.0, "open-ring indole diterpenoid compounds peniditerpenoid A derived from marine fungi, and preparation method and application thereof"; chinese patent application 3: CN202210093211.9, "chlororesorcinol aldehyde compound and its application in preparing osteoclast differentiation inhibitor", chinese patent application 4: CN201910176291.2, "a terpenoid derivative, its preparation method and application. The indole diketopiperazine compounds have no effect of resisting osteoclast differentiation, and no related application is reported. Therefore, more drugs which are safe, effective, controllable in quality and economical and can inhibit the formation of osteoclasts and bone resorption in a targeted way are further developed, the method has great significance in solving the clinical demands for osteoclast differentiation inhibitors, and meanwhile, the application value of the indole diketopiperazine compounds can be improved.
Disclosure of Invention
The invention aims at the problems and provides an indole diketopiperazine compound, a preparation method thereof and application thereof in preparing an osteoclast differentiation inhibitor.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
an indolyl diketopiperazine compound, designated prenylcyclotryprostatin B; the structural formula of prenylcyclotryprostatin B is shown as formula (I):
another object of the present invention is also to protect a strain of Penicillium brefeldianum (Penicillium brefeldianum) GXIMD 02511, deposited at the microorganism strain collection (GDMCC) of Guangdong province, month 09 of 2022, address: the Guangzhou City of Guangdong province, first, the middle road No. 100 Guangdong province microorganism research No. 59 building five, the Guangdong province institute of microorganisms, accession number: GDMCC No.62843.
The invention also aims at protecting a method for preparing the indole diketopiperazine compound, wherein the indole diketopiperazine compound is prepared and separated from a fermentation culture of the strain Penicillium brefeldianum (Penicillium brefeldianum) GXIMD 02511.
Further, the method comprises the following steps:
s1 preparation of fermentation culture: preparing a fermentation culture of the strain penicillium brefeldii (Penicillium brefeldianum) GXIMD 02511;
s2, extracting: soaking the fermentation culture in ethyl acetate, cutting or pulverizing the fermentation culture, performing ultrasonic extraction, and filtering to obtain filter residue and filtrate; extracting the filtrate and the filter residue with ethyl acetate respectively, concentrating to remove ethyl acetate, and mixing the extracts;
s3, separation and purification: subjecting the extract to medium pressure normal phase liquid chromatography with petroleum ether/dichloromethane as eluent, wherein the volume ratio is 100:0 to 0:10, gradient elution is carried out, and petroleum ether is collected: the volume ratio of dichloromethane is 90:10 fractions eluted in gradient, continuing through medium-pressure reversed-phase C 18 Column chromatography, using methanol/water as eluent, from a volume ratio of 10: 90-100: 0, gradient elution, collecting methanol: water volume ratio 30:70, purifying the eluted fraction to obtain the indole diketopiperazine compound.
Further, in S1, the fermentation culture is prepared by fermentation according to the following method: inoculating activated strain Penicillium brefeldianum (Penicillium brefeldianum) GXIMD 02511 into seed culture medium, and dynamically culturing at 25deg.C and 180rpm for 72 hr to obtain seed solution; inoculating the seed solution into a fermentation culture medium in an inoculum size of 5%, and statically culturing for 30 days at 25 ℃ to obtain a fermentation culture; the seed culture medium formula comprises the following components in each 1L of culture medium: 15g of malt extract powder, the balance of water and pH 7.5; the formula of the fermentation medium comprises the following components in each 1L of triangular flask medium: 200g of rice, 2g of sea salt, 200mL of water and pH 7.5.
The invention also aims at protecting the application of the indole diketopiperazine compound in preparing an osteoclast differentiation inhibitor.
Further, the osteoclast differentiation inhibitor is a medicament for treating osteolytic diseases caused by overactivation of osteoclasts.
Further, the osteoclast differentiation inhibitor is a medicament for treating osteoporosis, rheumatoid arthritis and tumor metastasis bone destruction.
The invention also aims at protecting the application of the indole diketopiperazine compound in preparing NF- κB nuclear factor expression inhibitor drugs.
It is a further object of the present invention to provide a pharmaceutical composition which may be an NF- κb nuclear factor expression inhibitor or an osteoclast differentiation inhibitor drug, comprising an effective amount of an indoledione piperazine compound and/or a pharmaceutically acceptable salt and a pharmaceutically acceptable carrier or adjuvant.
The invention is obtained by test: the indole diketopiperazine compound prenylcyclotryprostatin B has an inhibition effect (p < 0.001) on LPS-induced NF- κB luciferase at a concentration of 10 mu M, can be used as a lead compound for developing an NF- κB nuclear factor expression inhibitor, can remarkably inhibit RANKL-induced osteoclast precursor cells BMMs (Bone marrow macrophage cells, bone marrow macrophages) from differentiating into osteoclasts, and has no obvious cytotoxicity, so the indole diketopiperazine compound prenylcyclotryprostatin B is expected to be developed into a safe and effective novel osteoclast differentiation inhibitor drug.
By adopting the technical scheme, the invention has the following beneficial effects:
in the research process of secondary metabolites of penicillium brefeldianum (Penicillium brefeldianum) GXIMD 02511 which are the rhizosphere substrate sludge source in the protection zone of the mangrove forest in the northern Europe of Guangxi, the invention separates and obtains the indole diketopiperazine compound prenylcyclotryprostatin B which has obvious inhibition effect on NF- κB luciferase induced by LPS, obviously inhibits the differentiation of bone-breaking precursor cells BMMs induced by RANKL into bone-breaking cells in a dose-dependent manner, and has no cytotoxicity. Therefore, the compound is an ideal candidate compound developed into a novel NF- κB nuclear factor expression inhibitor or an osteoclast differentiation inhibitor drug.
Drawings
FIG. 1 is a graph showing comparison of inhibitory activity of indole diketopiperazine prenylcyclotryprostatin B against Lipopolysaccharide (LPS) -induced NF- κB luciferase in RAW264.7 cells at a concentration of 10 μM, BAY being a positive control, ### p<0.001vs.control group；***p<0.001vs.LPS group；
FIG. 2 is a graph showing the effect of indole diketopiperazine prenylcyclotryprostatin B on cell viability of mouse Bone Marrow Macrophages (BMMs);
fig. 3 is an experimental result of the effect of indole diketopiperazine prenylcyclotryprostatin B on differentiation of osteoclast precursor cells BMMs into osteoclasts, wherein: RANKL is a ligand for activating nuclear factor NF- κb receptor, and # # # P <0.001 compared to the blank; p <0.05, P <0.01 compared to RANKL group;
Detailed Description
In order to make the objects, technical schemes and technical effects of the present invention more clear, the present invention will be further described in detail with reference to the following examples and the accompanying drawings. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
Example 1
An indolyl diketopiperazine compound named prenylcyclotryprostatin B; the structural formula of prenylcyclotryprostatin B is shown as formula (I):
EXAMPLE 2 strain Penicillium brefeldianum (Penicillium brefeldianum) GXIMD 02511
The strain Penicillium brefeldianum (Penicillium brefeldianum) GXIMD 02511 was isolated from the rhizosphere bottom sludge of the mangrove protection area of the mouth of Heteropap, guangdong province, china, deposited at the microbiological culture Collection center (GDMCC) at 2022, 09, 27, address: the Guangzhou City of Guangdong province, first, the middle road No. 100 Guangdong province microorganism research No. 59 building five, the Guangdong province institute of microorganisms, accession number: GDMCC No.62843.
The strain Penicillium brefeldin (Penicillium brefeldianum) GXIMD 02511 of this example can be used to prepare indole diketopiperazine prenylcyclotryprostatin B of example 1.
EXAMPLE 3 preparation and isolation of indole diketopiperazine prenylcyclotryprostatin B
1. Culture medium
1.1, seed culture medium: each 1L of the culture medium contains 15g of malt extract powder, the balance of water and has the pH of 7.5. Mixing the above components and contents, and sterilizing at 121deg.C for 30 min.
1.2, fermentation medium: each 1L of the flask medium contains: 200g of rice, 2g of sea salt, 200mL of water and pH 7.5. Mixing the above components and contents, and sterilizing at 121deg.C for 30 min.
2. Fermentation
2.1, seed culture: the activated strain Penicillium brefeldianum (Penicillium brefeldianum) GXIMD 02511 was inoculated into 1L triangular flask containing 300mL of seed medium per flask, and cultured at 25℃and 180rpm for 72 hours to obtain a seed solution.
2.2, fermentation culture: the seed solution was inoculated into 120 flasks of fermentation medium at an inoculum size (volume percent) of 5%, and statically cultured at 25℃for 100 days to prepare a fermentation culture.
3. Extracting: soaking the fermentation culture in ethyl acetate with volume of 2 times for about 24 hours, cutting or mashing, ultrasonic extracting in an ultrasonic instrument for 20min, and filtering with 8 layers of gauze to obtain filtrate and residue respectively. The filtrate was concentrated by rotary evaporator to remove the organic solvent and then extracted with ethyl acetate multiple times until the color of the aqueous phase became light, and concentrated to remove ethyl acetate. The residue was extracted with ethyl acetate several times and concentrated to remove ethyl acetate, and the two brown yellow extractum were combined to about 132g.
4. Separation and purification of indole diketopiperazine compounds prenylcyclotryprostatin B
Subjecting the extract (132 g) to medium-pressure normal-phase liquid chromatography, using petroleum ether/dichloromethane as eluent, performing gradient elution from the volume ratio (100:0) - (0:100), and collecting petroleum ether/dichloromethane volume ratio of 90:10 fractions eluted in gradient, continuing through medium-pressure reversed-phase C 18 Column chromatography, using methanol/water as eluent, gradient elution from volume ratio (10:90) - (100:0), collecting methanol/water volume ratio 30:70, and finally separating the fraction by semi-preparative high performance liquid chromatography, purifying with methanol/water (volume ratio 50:50, YMC-pack ODS-A chromatographic column, 10×250mm,5 μm,2 m/min) to obtain indole diketopiperazine compound prenylcyclotryprostatin B (3 mg).
The structure of the indole diketopiperazine compound prenylcyclotryprostatin B obtained by the extraction and separation is identified prenylcyclotryprostatin B as white powder, 1 the H NMR spectrum of the compound can be deduced that the compound contains a 1,2, 4-trisubstituted benzene ring (. Delta.) H 7.46,6.81,6.74), 4 methyl groups (. Delta.) H 2.14,2.00,1.83,1.76) 2 methoxy groups (. Delta H 3.84,3.67), and signals for 4 methylene and methylamino groups. 13 The C NMR spectrum showed 28 carbon signals including 6 methyl groups, 3 aromatic carbons, 4 methylene groups, 5 methine groups and 10 aprotic carbons. The nuclear magnetic data are attributed as follows:
1 H NMR(700MHz,CDCl 3 )δ H 7.46(1H,dd,J＝8.6,2.7Hz,H-4),6.81(1H,dd,J＝8.6,2.2Hz,H-5),6.74(1H,d,J＝2.2Hz,H-7),6.73(1H,d,J＝9.7Hz,H-18),5.56(1H,m,H-19),5.12(1H,d,J＝5.9Hz,H-24),4.72(1H,s,H-8),4.62(1H,dd,J＝16.0,6.4Hz,H-23a),4.52(1H,dd,J＝16.0,6.4Hz,H-23b),4.48(1H,br s,9-OH),4.41(1H,dd,J＝10.8,6.2Hz,H-12),3.84(3H,s,6-OCH 3 ),3.75(2H,m,H 2 -15),3.67(3H,s,8-OCH 3 ),2.49(1H,m,H-13a),2.14(1H,m,H-14a),2.00(3H,s,H 3 -22),1.98(1H,m,H-13b),1.98(1H,m,H-14b),1.83(3H,s,H 3 -27),1.76(3H,s,H 3 -21),1.71(3H,s,H 3 -26)； 13 C NMR(175MHz,CDCl 3 )δ C 165.7(C-11),164.7(C-17),156.8(C-6),138.2(C-20),137.9(C-7a),134.9(C-25),133.4(C-2),123.9(C-19),120.4(C-3a),120.0(C-24),118.7(C-4),109.9(C-5),104.5(C-3),94.5(C-7),85.7(C-9),77.2(C-8),60.2(C-12),59.2(OCH 3 -8),55.9(OCH 3 -6),48.3(C-18),45.9(C-15),42.3(C-23),30.0(C-13),26.2(C-21),25.6(C-26),22.04(C-14),18.62(C-22),18.35(C-27).
the above data are essentially identical to those reported in literature (Chemistry & Biodiversity,2012, 9:385-393.) and therefore identify isopentenyl indole diketopiperazine compounds prenylcyclotryprostatin B.
On the basis of the above, the invention also carries out the following experiment:
experiment one: determination of LPS-induced NF- κB luciferase inhibition activity of indole diketopiperazine prenylcyclotryprostatin B
NF- κB luciferase inhibition activity assay primary reference (Fisterapia, 2022, 159:105201.).
RAW264.7 cells stably transfected with NF- κB luciferase reporter gene were inoculated in 96-well plates (1×10) 4 200. Mu.L of DMEM medium containing 10% fetal bovine serum, 100IU/mL penicillin and streptomycin and 0.1. Mu.g/mL G418 was added to each well, and after cell attachment was stabilized, indole diketopiperazine prenylcyclotryprostatin B was added to set 6 multiplex wells. After further incubation for 4h, LPS and RANKL were added to each of the compound group (3 wells) and positive control group (NF- κB inhibitor, BAY11-7082,5. Mu.M) respectively to a final concentration of 100ng/mL per well, after 8h of stimulation, the supernatant was discarded and cell lysate 25 was added per wellmu.L, shake at low speed for 10min to lyse the cells thoroughly, transfer 20. Mu.L to the whiteboard, add 50. Mu.L of fluorescein solution per well, and check the Lucifer value with a multifunctional microplate reader.
Conclusion of the test: as shown in FIG. 1, the indole diketopiperazine prenylcyclotryprostatin B has a moderate inhibition (p < 0.001) of LPS-induced NF- κB luciferase at 10 μM compared to the LPS blank.
Experiment II: effect of indolyl diketopiperazine prenylcyclotryprostatin B on the viability of osteoclast precursor cells BMMs
Method for detecting activity influence of indole diketopiperazine prenylcyclotryprostatin B on mouse Bone Marrow Macrophages (BMMs) by CCK-8 method
a. Preparation of mouse Bone Marrow Macrophages (BMMs): under the aseptic condition, taking femur of C57BL/6 female mice with age of 8-12 weeks, cutting joint parts at two ends of femur, repeatedly flushing femur with phenol red-free alpha-MEM culture medium (containing 10% fetal calf serum, 100IU/mL penicillin and 100IU/mL streptomycin) until femur cavity blurs. Placing the washed bone marrow cavity cells of femur at 37deg.C and 5% CO 2 Incubating the cells in a cell incubator for 2 hours, absorbing the supernatant, lysing the red blood cells by using lysate, centrifuging and re-suspending to obtain the BMMs.
CCK-8 method to detect cell survival:
taking the BMMs (1×10) prepared in step a 5 0/well) was added to 200. Mu.L of phenol red-free alpha-MEM medium (containing 10% fetal bovine serum, 100IU/mL penicillin and 100IU/mL streptomycin) per well while macrophage colony stimulating factor (M-CSF, final concentration 50 ng/mL) was added per well, and then the 96-well plate was placed at 37℃with 5% CO 2 Is incubated overnight. After the cell adhesion is stable, adding the indole diketopiperazine compounds prenylcyclotryprostatin B with different concentrations respectively, so that the final concentration of the indole diketopiperazine compounds prenylcyclotryprostatin B in the holes is 10 mu M and 15 mu M, and each group is provided with 3 compound holes for 4 days of incubation. After the incubation was completed, the supernatant (100. Mu.L) was discarded, 5. Mu.L of CCK-8 reagent (Cell Counting Kit-8 cell counting reagent) was added to each well, and the mixture was shaken and placed at 37℃in 5% CO 2 In the environmentIncubation was continued for 3h, and the Optical Density (OD) at 450nm was measured using a TECANGENiosPro multifunctional microplate reader, and the cell viability of each group was calculated.
The results, as shown in figure 2, show that there was no significant difference in BMMs cell viability after addition of 10 μm and 15 μm of indole diketopiperazine prenylcyclotryprostatin B, indicating that indole diketopiperazine prenylcyclotryprostatin B was non-cytotoxic to BMMs in vitro assays.
Experiment III: effect of indole diketopiperazine prenylcyclotryprostatin B on RANKL-induced differentiation of osteoclast precursor cells BMMs into osteoclasts
RANKL-induced osteoclast precursor BMMs cell differentiation inhibitory activity was determined as the primary reference (fitterapia, 2022, 159:105201.).
Taking BMMs (2×10) obtained in step a above 4 0/well) was added to 200. Mu.L of phenol red-free alpha-MEM medium (containing 10% fetal bovine serum, 100IU/mL penicillin and 100IU/mL streptomycin) per well while macrophage colony stimulating factor (M-CSF, final concentration 50 ng/mL) was added per well, and then the 96-well plate was placed at 37℃with 5% CO 2 Is incubated overnight. After the cell adhesion is stable, adding the indole diketopiperazine compounds prenylcyclotryprostatin B with different concentrations respectively, so that the final concentration of the indole diketopiperazine compounds prenylcyclotryprostatin B in the holes is 5 mu M,10 mu M and 15 mu M, and each group is provided with 3 compound holes for 4 hours of incubation. After the incubation, RANKL (final concentration 100 ng/mL) was added and incubated for 3-4d. TRAP staining is carried out on the cells after incubation, and the cells are photographed and counted under an inverted microscope, wherein TRAP positive cells with more than 5 nuclei are osteoclasts.
As shown in fig. 3, the indole diketopiperazine compound prenylcyclotryprostatin B can significantly inhibit RANKL from inducing BMMs to generate osteoclasts. At the effective concentration of 10 mu M, the preparation can obviously inhibit the RANKL from inducing the osteoclast precursor cells BMMs to generate osteoclasts.
From the above results, it is known that the indole diketopiperazine compound prenylcyclotryprostatin B has a remarkable inhibitory effect on NF- κb luciferase induced by LPS, can remarkably inhibit RANKL-induced differentiation of osteoclast precursor cells BMMs into osteoclasts at a concentration of 10 μm, can remarkably inhibit generation and activation of osteoclasts in vitro, has no remarkable toxic effect, can be developed as a novel osteoclast differentiation inhibitor or NF- κb nuclear factor expression inhibitor, and is used for preventing and treating bone-soluble diseases such as osteoporosis.
Based on the experiment, the application of the two indole diketopiperazine compounds prenylcyclotryprostatin B in preparing the anti-osteoclast differentiation inhibitor or NF- κB nuclear factor expression inhibitor medicine can be proved. The osteoclast differentiation inhibitor or NF- κB nuclear factor expression inhibitor is in the form of oral preparation, injection or external preparation; comprises an active ingredient of indole diketopiperazine compound prenylcyclotryprostatin B and medical acceptable pharmaceutic adjuvant. Anti-osteoclast differentiation class drugs may be treated including, but not limited to: clinical indications for which known osteoclast inhibitors such as biphosphate and denoximab are approved for treatment, such as postmenopausal osteoporosis and tumor metastasis bone destruction.
Generally, the medicines are clinically applied after being prepared into preparations. The indole diketopiperazine compound serving as an effective active ingredient can be prepared according to a method known in the art. Any dosage form suitable for human or animal use can be made by combining the active ingredient indole diketopiperazine compounds of the invention with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants. The indole diketopiperazine compound as an effective active ingredient or the indole diketopiperazine compound containing the same can be administered in a unit dosage form, and the administration route can be intestinal tract or parenteral tract, such as oral administration, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eye, lung, respiratory tract, skin, vagina, rectum and the like.
The oral preparation is preferably capsule. In order to prepare the administration unit into a capsule, the phenylacetic acid compound serving as an effective ingredient of the present invention may be mixed with a diluent and a glidant, and the mixture may be directly placed into a hard capsule or a soft capsule. The indole diketopiperazine compound as the effective component can be prepared into particles or pellets by mixing the effective component with a diluent, an adhesive and a disintegrating agent, and then placing the particles or pellets into hard capsules or soft capsules. The diluents, binders, wetting agents, disintegrants and glidants used for preparing the tablets of the indole diketopiperazine compounds serving as the effective active ingredients can also be used for preparing capsules of the indole diketopiperazine compounds serving as the effective active ingredients.
In order to prepare the indole diketopiperazine compound serving as an effective active ingredient into an injection, water, ethanol, isopropanol, propylene glycol or a mixture thereof can be used as a solvent, and a proper amount of a solubilizer, a cosolvent, a pH regulator and an osmotic pressure regulator which are commonly used in the field can be added. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-beta-cyclodextrin, etc.; the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. For example, mannitol, glucose, etc. can be added as propping agent for preparing lyophilized powder for injection.
In summary, the invention provides a new candidate compound for developing a novel osteoclast differentiation inhibitor or NF- κB nuclear factor expression inhibitor drug, and has important significance for developing new drugs with independent intellectual property rights.
The above description is of a detailed description of a preferred embodiment of the invention, but the embodiment is not intended to limit the scope of the invention. All equivalent changes or modification changes which are accomplished under the technical conception suggested by the invention are included in the scope of the patent covered by the invention.

Claims (10)

1. The indole diketopiperazine compound is characterized in that the indole diketopiperazine compound is named prenylcyclotryprostatin B; the structural formula of prenylcyclotryprostatin B is shown as formula (I):
2. a strain of penicillium brefeldii (Penicillium brefeldianum) GXIMD 02511 for the preparation of an indolyl diketopiperazine compound of claim 1, deposited under the accession number: GDMCC No.62843.
3. A process for the preparation of the indole diketopiperazine compound according to claim 1, characterized in that the indole diketopiperazine compound is isolated from a fermentation culture of the strain penicillium brefeldianum (Penicillium brefeldianum) GXIMD 02511.
4. A method according to claim 3, characterized in that the method comprises the steps of:
s1 preparation of fermentation culture: preparing a fermentation culture of the strain penicillium brefeldii (Penicillium brefeldianum) GXIMD 02511;
s2, extracting: soaking the fermentation culture in ethyl acetate, cutting or pulverizing the fermentation culture, performing ultrasonic extraction, and filtering to obtain filter residue and filtrate; extracting the filtrate and the filter residue with ethyl acetate respectively, concentrating to remove ethyl acetate, and mixing the extracts;
s3, separation and purification: subjecting the extract to medium pressure normal phase liquid chromatography with petroleum ether/dichloromethane as eluent, wherein the volume ratio is 100:0 to 0:10, gradient elution is carried out, and petroleum ether is collected: the volume ratio of dichloromethane is 90:10 gradient eluted fraction, and the eluted fraction is continuously passed through medium-pressure reversed phase C 18 Column chromatography, using methanol/water as eluent, from a volume ratio of 10: 90-100: 0, gradient elution, collecting methanol: water volume ratio 30:70, purifying the fractions eluted in a gradient way to obtain the indole diketopiperazine compound.
5. The method of claim 4, wherein in S1, the fermentation culture is prepared by fermentation by: inoculating activated strain Penicillium brefeldianum (Penicillium brefeldianum) GXIMD 02511 into seed culture medium, and dynamically culturing at 25deg.C and 180rpm for 72 hr to obtain seed solution; inoculating the seed solution into a fermentation culture medium in an inoculum size of 5%, and statically culturing for 30 days at 25 ℃ to obtain a fermentation culture; the seed culture medium formula comprises the following components in each 1L of culture medium: 15g of malt extract powder, the balance of water and pH 7.5; the formula of the fermentation medium comprises the following components in each 1L of triangular flask medium: 200g of rice, 2g of sea salt, 200mL of water and pH 7.5.
6. The use of an indole diketopiperazine compound according to claim 1 for the preparation of an osteoclast differentiation inhibitor.
7. The use according to claim 6, characterized in that: the osteoclast differentiation inhibitor is a medicine for treating osteolytic diseases caused by overactivation of osteoclasts.
8. The use according to claim 6, characterized in that: the osteoclast differentiation inhibitor is a medicament for treating osteoporosis, rheumatoid arthritis and tumor metastasis bone destruction.
9. The use of an indole diketopiperazine compound according to claim 1 for the preparation of an NF- κb nuclear factor expression inhibitor drug.
10. A pharmaceutical composition comprising an effective amount of prenylcyclotryprostatin B and/or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or adjuvant.
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