CN111840561A - Application of S100A9 inhibitor in preparation of medicine for treating pancreatitis - Google Patents
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- 206010033645 Pancreatitis Diseases 0.000 title claims abstract description 30
- 239000003814 drug Substances 0.000 title claims abstract description 13
- 238000002360 preparation method Methods 0.000 title abstract description 3
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- 206010033647 Pancreatitis acute Diseases 0.000 claims description 4
- 201000003229 acute pancreatitis Diseases 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 2
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- 230000000694 effects Effects 0.000 abstract description 4
- 238000010874 in vitro model Methods 0.000 abstract description 3
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- 238000002474 experimental method Methods 0.000 description 5
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- JAJWGJBVLPIOOH-IZYKLYLVSA-M sodium taurocholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 JAJWGJBVLPIOOH-IZYKLYLVSA-M 0.000 description 5
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- 125000000539 amino acid group Chemical group 0.000 description 2
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
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- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
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- 102000014823 calbindin Human genes 0.000 description 1
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- 230000034994 death Effects 0.000 description 1
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- 230000001988 toxicity Effects 0.000 description 1
- 238000011277 treatment modality Methods 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/425—Thiazoles
- A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
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Abstract
The invention discloses an application of an S100A9 inhibitor in preparation of a medicine for treating pancreatitis, and belongs to the technical field of biomedicine. Use of an S100a9 inhibitor, such as Chemdiv ID:0884, 3948, 1191,3232, 0780. The invention adopts an S100A9 inhibitor to pretreat an pancreatitis in vitro model, and proves that an S100A9 inhibitor (ID:0884-0014,3948-1191,3232-0780) can improve the activity of HPDE6-C7 cells and reduce STC-induced cell damage. The invention provides a basis for using the S100A9 inhibitor as a novel medicine for treating pancreatitis.
Description
Technical Field
The invention relates to an application of an S100A9 inhibitor in preparing a medicine for treating pancreatitis, belonging to the technical field of biomedicine.
Background
Acute pancreatitis is a common clinical local inflammation of pancreas, and is a digestive system disease mainly characterized by local necrosis and Multiple Organ Dysfunction (MODS), and the death rate is high at the head of various acute abdominal diseases. Currently, a non-operative treatment system mainly based on early supportive treatment is formed for acute pancreatitis, but due to insufficient understanding of pathogenesis of the disease, no specific preventive and therapeutic medicine exists so far. Clinical observations of patients with pancreatitis have shown that the pancreatic duct is most vulnerable to the noxious stimuli of cholelithiasis and endoscopic retrograde pancreatography, and the pancreatic duct is the tissue most vulnerable to the common noxious stimuli of pancreatitis. Therefore, it has been shown that the research on the pathophysiological changes after the catheter injury in pancreatitis is sufficiently carried out in order to search for better drug treatment means.
S100a9 is a member of the calbindin S100 protein family and is expressed predominantly on early differentiated cells of myeloid origin. The research shows that the S100A9 protein passes through Ca2+The heterodimer is formed in a dependent mode and plays a biological role in regulating physiological and pathological processes such as calcium balance in vivo, apoptosis, inflammatory reaction and the like through interaction with effector target proteins. However, the relationship between S100A9 and pancreatitis is rarely reported in the literature at present.
Disclosure of Invention
In order to solve the problems, the invention screens out 8S 100A9 inhibitors with the highest scores based on a drug target library (Chemdiv), detects the cytotoxicity of the inhibitors on a human normal pancreatic ductal epithelial cell strain HPDE6-C7 by adopting an MTT method, and screens out 3 drugs which can effectively treat pancreatitis by adopting an in-vitro pancreatitis model.
The invention provides an application of an S100A9 inhibitor in preparing a medicine for treating pancreatitis, wherein the S100A9 inhibitor is shown as a formula I, a formula II or a formula III;
further, in the above technical scheme, the pancreatitis is acute pancreatitis.
Further, in the above technical solution, the S100a9 inhibitor is an S100a9-VNN1 inhibitor.
A medicament comprising an inhibitor of S100a9 and pharmaceutically acceptable salts thereof for the treatment of pancreatitis.
According to the invention, the protein level of S100A9 is found to be obviously up-regulated in pancreatitis ductal tissues through research, and S100A9shRNA interference plasmids and gene knockout mice can obviously reduce in vitro and in vivo pancreatitis injuries; and its mechanism of action involves S100a9 up-regulating VNN 1-mediated ROS release levels. Therefore, inhibition of the S100a9-VNN1 interaction is one potential treatment modality for improving pancreatitis. Based on the molecular docking technology and the pancreatitis in-vitro model, three efficient S100A9-VNN1 inhibitors (ID: 0884-.
Advantageous effects of the invention
The invention adopts an S100A9 inhibitor to pretreat an pancreatitis in vitro model, and proves that an S100A9 inhibitor (ID:0884-0014,3948-1191,3232-0780) can improve the activity of HPDE6-C7 cells and reduce Sodium Taurocholate (STC) -induced cell damage. The invention provides a basis for using the S100A9 inhibitor as a novel medicine for treating pancreatitis.
Drawings
FIG. 1 is a binding model of S100A9 and VNN 1; (A) s100a9 interacted with the surface of VNN 1; (B) three-dimensional interaction of S100a9 with VNN 1.
FIG. 2 is a schematic representation of 8 virtually screened S100A9-VNN1 interaction inhibitors.
FIG. 3 shows the toxicity test of S100A9 inhibitor.
FIG. 4 shows drug screening of S100A9 inhibitor.
Fig. 5 is a graph showing that the S100a9 inhibitor improves STC-induced cell damage.
Detailed Description
The following non-limiting examples will allow one of ordinary skill in the art to more fully understand the present invention, but are not intended to limit the invention in any way.
Example 1
The first experimental scheme is as follows:
1. molecule docking: the 3D structures of the S100A9 protein and VNN1 protein were downloaded from the RCSB protein database (PDB ID:5I8N for S100A9 and 4CYF for VNN 1). Performing molecular docking simulation and predicting the binding affinity of the complex by utilizing the protein-protein docking function in the ClusPro; the docking structures and interfacial residues were analyzed using MOE v2018.01 to finally generate a molecular pattern from PyMOL, as shown in fig. 1.
2. Virtual screening: the Dock module in MOE v2015.1001 was used for receptor-based virtual screening (SBVS). The S100a9 protein was defined as the receptor and 10 ten thousand compounds of Chemdiv were defined as the virtual screening library. Receptor binding sites were selected near the residues where S100a9 interacted with VNN1, and the top 8 ranked inhibitors were finally selected, the structural formulae of the 8 inhibitors being shown in figure 2.
3. Toxicity test: the toxicity of 8S 100a9 inhibitors on HPDE6-C7 cells (human normal pancreatic ductal epithelial cells, available from QCB429, shanghai sinceri biotechnology, inc.) was determined by MTT method, as follows: the cells were cultured at 1X 105Each/ml (100. mu.l) was inoculated into a 96-well plate using DMEM medium containing 10% fetal bovine serum in 5% CO2After incubation for 24h at 37 ℃ under saturated humidity conditions, different concentrations (final concentrations of 10,1,0.1,0.01,0.001 and 0. mu.M) of S100A9 inhibitor (dissolved in DMSO, final concentration)<0.1%) for 24 h. Subsequently, 10. mu.l of MTT (5mg/ml) was added and incubated at 37 ℃ in the dark, after 4 hours, the MTT was blotted, 100. mu.l of DMSO was added, dissolved with shaking for 10 minutes, and the absorbance A490 was measured with a microplate reader. And calculating the collected data to obtain the cell viability inhibition rates of different groups, and screening the maximum non-toxic dose for subsequent drug effect research. Duplicate wells were set for each concentration, and each set of experiments was repeated 3 times. The results of the experiment are shown in FIG. 3.
4. And (3) pharmacodynamic experiment: HPDE6-C7 cells at 1X 105Each/ml (100. mu.l) was inoculated into a 96-well plate using DMEM medium containing 10% fetal bovine serum in 5% CO2After culturing for 24 hours under the condition of saturated humidity at 37 ℃, the cells are divided into 10 groups: 1) to pairGroup control; 2) a model group; 3) group 8S 100a9 inhibitor groups ( compounds 1, 2, 3, 4, 5, 6, 7 and 8). S100A9 inhibitor component maximum non-toxic doses (1, 0.001, 1,0.001 and 0.001. mu.M) of S100A9 inhibitor (DMSO dissolved at final concentration) were added<0.1%); the control group and the model group were added with the same volume of DMSO (final concentration)<0.1%). After 24h, adding 1015 mu M sodium taurocholate (STC, DMEM dissolved) solution into the cells of the model group and the 8 groups of S100A9 inhibitor groups to act for 1h, not processing the control group, finally detecting the cell viability by adopting an MTT method, and screening S100A9 inhibitors possibly used for treating pancreatitis. Each concentration was set to 6 multiple wells, each set of experiments was repeated 3 times, and the results are shown in fig. 4. And the cell damage was observed under an inverted microscope and photographed, and the experimental results are shown in fig. 5.
II, experimental results:
1. molecule docking: the model of the binding of S100A9 and VNN1 to each other is shown in FIG. 1. Orange structure represents S100a9, green structure represents VNN 1; the red dotted line represents hydrogen bonds and the blue dotted line represents salt bridges. Docking simulation studies have shown that residues His228, Arg259, Ser309, His310, Ser311, Val313, Val314, Asn315, Ser321, Ile323, Glu324, Phe431 and Gln434 in VNN1 bind to residues Glu52, Lys57, Arg85, Glu92, Met94, Gly100, Gly102, His103 and His105 in S100a9 through salt bridges and hydrogen bonding interactions. Specific docking results are shown in table 1.
TABLE 1 list of interacting amino acid residues of S100A9 and VNN1
2. Virtual screening: 8 inhibitors are screened according to the amino acid residues of the mutual combination of S100A9 and VNN1, and the structural formulas of the 8 inhibitors are shown in figure 2.
3. Toxicity test: as shown in FIG. 3, it was found that the maximum non-toxic doses of the 8 inhibitors (ID Nos: 0683-0021, 0884-0014, 2372-3991, 3948-1149, 3948-1191, 0249-0003, 2324-0140 and 3232-0780) were 1,0.001, 1, 0.001. mu.M, respectively.
4. And (3) pharmacodynamic experiment: MTT results show that compared with a model group, 3S 100A9 inhibitors (ID:0884-0014,3948-1191,3232-0780) can significantly improve the activity of HPDE6-C7 cells (FIG. 4), and meanwhile, the 3 inhibitors can improve cell damage and reduce cell debris when observed in an inverted microscope bright field (FIG. 5).
Claims (4)
1. The application of an S100A9 inhibitor in preparing a medicine for treating pancreatitis is characterized in that the S100A9 inhibitor is shown as a formula I, a formula II or a formula III;
2. the use according to claim 1, wherein said pancreatitis is acute pancreatitis.
3. The use of claim 1, wherein the S100a9 inhibitor is a S100a9-VNN1 inhibitor.
4. A medicament for the treatment of pancreatitis, comprising an inhibitor of S100a9 and pharmaceutically acceptable salts thereof.
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Cited By (2)
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| CN115120727A (en) * | 2022-06-16 | 2022-09-30 | 甘肃农业大学 | Application of S100A9 inhibitor in preparation of drugs for preventing and treating C-type clostridium perfringens infectious diarrhea |
| WO2023141659A3 (en) * | 2022-01-24 | 2023-09-14 | The Trustees Of Columbia University In The City Of New York | Targeting s100a9-aldh1a1-retinoic acid signaling to suppress brain relapse in egfr-mutant lung cancer |
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