CN110787166A

CN110787166A - Small molecule compound C28H26Cl2N8Application in E.coli meningitis

Info

Publication number: CN110787166A
Application number: CN201911074714.6A
Authority: CN
Inventors: 陈誉华; 徐晓倩; 赵伟东
Original assignee: China Medical University
Current assignee: China Medical University
Priority date: 2019-11-06
Filing date: 2019-11-06
Publication date: 2020-02-14
Anticipated expiration: 2039-11-06
Also published as: CN110787166B

Abstract

The invention discloses a small molecule compound C₂₈H₂₆Cl₂N₈The compound or an isomer, a solvate or a precursor thereof, or pharmaceutically acceptable salts thereof can be combined with hot spot amino acid residues of interaction of a brain vascular endothelial receptor protein Caspr1 in a bacterial meningitis virulence factor IbeA protein in a targeted competitive manner so as to block the combination of the two, thereby achieving the purpose of preventing the escherichia coli from passing through a blood brain barrier or blocking the meningitis escherichia coli from invading neurons. The invention starts from the aspects of intervening the interaction of a ligand (IbeA) -receptor (Caspr1) of a bacterium-host and blocking pathogenic bacteria, adopts a new targeted treatment method and is the bacterial meningitis of the newbornProvides a new target medicine for preventing and treating.

Description

Small molecule compound C28H26Cl2N8Application in E.coli meningitis

Technical Field

The invention relates to the technical field of medicaments, in particular to a new medicament target for blocking a pathogenic virulence factor IbeA of neonatal meningitis from passing through a blood brain barrier and attacking neurons, and application of the new medicament target in preparing new medicaments for the neonatal meningitis.

Neonatal bacterial meningitis is one of serious infectious diseases in pediatrics, and clinically, despite antibiotics and related supportive therapies against bacteria, 5% -40% of infants die due to bacterial drug resistance and difficulty in passing through barrier structures between blood and brain tissues of most antibiotics. Escherichia coli (e.coli) K1 strain is the most common gram-negative pathogen causing bacterial meningitis in newborns. The primary condition for causing e.coli meningitis is that bloodborne e.coli must cross the blood-brain barrier, which is mainly composed of Brain Microvascular Endothelial Cells (BMECs), where the key link is e.coli adhesion and invasion into BMECs. Recently, in the laboratory of the applicant, neuron contact chaperonin 1 (Caspr1) is identified in vivo as a receptor of Escherichia coli virulence factor IbeA on BMECs, and the IbeA is combined with a Laminin protein family domain consisting of Caspr1 extracellular segment 203-355aa through 229-343 amino acid fragment to realize the penetration of the blood brain barrier of bacteria. Based on the above research results, we first constructed the three-dimensional spatial structures and interaction sites of Caspr1 (203-. The presence of a stable small molecule binding pocket in IbeA (229-.

In view of the fact that the Caspr1 and IbeA protein purification cannot meet the technical requirements of structure analysis, the subject group of the applicant constructs three-dimensional space structures of Caspr1 (203) -355aa) and IbeA (229-343aa) by a computer-assisted homologous modeling method, predicts a binding mode and an interaction site between the Caspr1 (203) -355aa) and the IbeA (229-343aa), and determines stable binding of the Caspr1 (203) -355aa) and the IbeA (229-343aa) through MM/GBSA and RMSD. Based on the prediction model, the inventor finds that IbeA (229-.

CN108611411A discloses the use of Caspr1 membrane protein as a new drug for preparing neonatal meningitis drugs, this patent application discloses the use of Caspr1 membrane protein or IbeA-Caspr1 interacting domain as a new drug target for blocking meningitis escherichia coli from crossing the blood brain barrier and attacking neurons and as a new drug for preparing neonatal meningitis drugs, and discloses the preparation of oligopeptide drugs for blocking meningitis escherichia coli from crossing the blood brain barrier based on Caspr 1203-355 peptide fragment, but this technical scheme performs the knockout of Caspr1 gene on rats, thereby significantly reducing the incidence of neonatal mouse meningitis caused by e.coli K1, which is impractical in actual clinical and drug preparation, and the knockout of patient gene can cause a large number of problems that are difficult to verify subsequently.

In view of the above, the present invention is particularly proposed.

Disclosure of Invention

The invention aims to provide a small molecule compound C₂₈H₂₆Cl₂N₈The application in preparing the medicine for treating the E.coli meningitis.

In order to achieve the purpose, the invention adopts the following technical scheme:

small molecule compound C₂₈H₂₆Cl₂N₈The application in preparing the medicine for treating the E.coli meningitis;

the small molecule compound C₂₈H₂₆Cl₂N₈The structural formula of (A) is shown as formula I:

furthermore, the compound shown in the formula I, or an isomer, a solvate or a precursor thereof, or a pharmaceutically acceptable salt thereof is combined with hot spot amino acid residues of interaction between the bacterial meningitis virulence factor IbeA protein and the brain vascular endothelial receptor protein Caspr1 in a targeted and competitive manner, so that the combination of the two is blocked, and the purpose of preventing escherichia coli from passing through a blood brain barrier or blocking the meningitis escherichia coli from invading neurons is achieved.

Further, the IbeA protein is used as a virulence factor of pathogen E.coli K1 of neonatal meningitis.

Further, the interaction hotspot amino acid residues of the IbeA-Caspr1 are R189, W196, R331, V332, N333, L337, S202, G325, I328, F329, N330, D336, R340, H342 and R344 derived from Caspr1; v255, L319, F322, G323, T325, V326, G259, L261, T265, E266, M324, R329, S331 are derived from IbeA.

Furthermore, the medicine for treating the meningitis of the escherichia coli is used for blocking pathogenic bacteria to block meningitis escherichia coli from crossing a blood brain barrier or blocking meningitis escherichia coli from invading neurons.

Furthermore, the medicine for treating the E.coli meningitis is a medicine for treating central nervous system diseases which can easily pass through a blood brain barrier.

Furthermore, the medicine for treating the E.coli meningitis is a small molecular compound medicine.

A pharmaceutical composition for treating e.coli meningitis, comprising: a compound of formula (I) or an isomer, solvate or precursor thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier;

formula I:

a pharmaceutical composition for treating e.coli meningitis, comprising: a compound of formula (I) or an isomer, solvate or precursor thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier; and other effective components for treating E.coli meningitis.

Formula I:

furthermore, the other effective components for treating the E.coli meningitis are broad-spectrum antibiotics.

Further, in the pharmaceutical composition, the pharmaceutically acceptable carrier is a pharmaceutically or comestibly acceptable solvent, suspending agent or excipient for delivering the compound of formula I, isomer, solvate, precursor, or pharmaceutically acceptable salt thereof of the present invention to an animal or human; the carrier may be a liquid or a solid. .

The invention discloses an interaction site of a virulence factor IbeA of bacterium E.coli K1 and a receptor Caspr1 on human brain microvascular endothelial cells, so that a small molecule compound medicine for preventing and treating neonatal bacterial meningitis, which blocks the interaction, is screened and prepared on the basis of an IbeA-Caspr1 interaction domain.

In another aspect of the invention, a structural model of the interaction between Caspr1(203-355 amino acid peptide fragment) and IbeA (229-343 amino acid peptide fragment) was constructed for the first time, and the interaction amino acid hot spot residues were reported. Wherein the hot spot residues in IbeA (229-343 peptide segment) are mainly concentrated on 1 and 3 helices and form a small molecule binding pocket at the periphery, which is closely related to bacterial invasion.

In another aspect of the invention, IbeA (229-343 peptide fragment) is used as a target for the first time, and the obtained molecular model is virtually screened from a drug-like small molecular compound library to search for potential inhibitors from 213279 commercially available compounds. The first 50 small molecules with ordered binding energy were tested in vitro by Surface Plasmon Resonance (SPR) for their binding capacity titration to IbeA (229-343aa) purified protein, and 9 compounds showed direct binding capacity. Furthermore, in vitro toxicity tests and bacterial invasion experiments prove that the 4 molecules can obviously reduce or prevent the invasion of bacteria into human brain microvascular endothelial cells (hBMECs) within a low cytotoxicity range.

Compared with the prior art, the invention has the beneficial effects that:

(1) the prior art has no effective method aiming at preventing the occurrence of the neonatal bacterial meningitis, the neonatal meningitis is treated by adopting broad-spectrum antibiotics, and the defect that 5 to 40 percent of the sick children still die due to bacterial resistance and the fact that most of the antibiotics are difficult to pass through a barrier structure between blood and brain tissues; and the principle of antibiotic treatment is to kill bacteria, but at the same time is associated with a large amount of inflammatory reactions, such as hyperthermia, so that even if cured, 30% of the surviving children remain with permanent neurological sequelae, such as epilepsy, mental and movement disorders, etc. In this context, it is of great interest to develop targeted drugs that are effective in preventing bacterial meningitis in newborn infants.

(2) The invention provides a novel targeted medicine for preventing and treating the bacterial meningitis of the newborn by adopting a novel targeted treatment method from the viewpoints of intervening the interaction of a ligand (IbeA) -receptor (Caspr1) of a bacterium-host and blocking pathogenic bacteria.

Drawings

FIG. 1 is a diagram showing the interaction structure pattern of Caspr1 (203) -355 amino acid peptide fragment) and IbeA (229-343 amino acid peptide fragment) and hot spot amino acid residues, wherein A is the hot spot amino acid residue in the Caspr1 (203) -355aa) -IbeA (229-343aa) complex for maintaining stable structure, and the key site sequence and the amino acid name are marked with orange characters at the corresponding sites; b is a Caspr-IbeA interaction pattern diagram, and key amino acid sites are marked by black characters; c is a surface structure pattern diagram of an IbeA (229-343) amino acid peptide segment, and orange color shows a small molecule binding pocket related to bacterial invasion.

FIG. 2 is a graph showing the affinity of Top50 small molecules to IbeA protein screened by SPR in vitro test SPECS database, wherein 9 small molecules show the characteristic of direct binding to IbeA (229-343 aa).

FIG. 3 is a diagram of toxicity of small molecules capable of directly binding to IbeA in human brain microvascular endothelial cells (hBMEC) detected by CCK-8 assay.

Fig. 4 is a graph of bacterial invasion experiments to verify the blocking effect of 5 low-toxicity small-molecule compounds on the invasion of escherichia coli e.coli K1 into human brain microvascular endothelial cells.

FIG. 5 is a schematic diagram of the docking structure of a small molecule compound and IbeA (229-343 amino acid peptide) for blocking E.coli K1 from invading human brain microvascular endothelial cells, wherein A is C₂₈H₂₆Cl₂N₈(AK-968/11283147) and IbeA (229-; b is C₂₉H₂₈Cl₂N₆The interaction pattern diagram of O (AE-641/30154051) and IbeA (229-; c is C₃₄H₂₂FN₃O₅S₂(AF-399/42017932) and IbeA (229-; d is C₂₉H₂₂N₂O₈(AN-652/14563011) and IbeA (229-.

Detailed Description

The inventor of the invention, through the intensive research, reveals a small molecule compound C for the first time₂₈H₂₆Cl₂N₈The application of the polypeptide in preparing the medicine for treating the meningitis of the escherichia coli can block the meningitis escherichia coli from passing through a blood brain barrier or blocking the meningitis escherichia coli from invading neurons by targeting and competitively combining meningitis virulence factor IbeA protein or IbeA-Caspr1 interaction hot spot amino acid residues. The present invention has been completed based on this finding.

In the present invention, a "pharmaceutically acceptable" component is a substance that is suitable for use in humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic response) commensurate with a reasonable benefit/risk ratio.

In the present invention, the "pharmaceutically acceptable carrier" is a pharmaceutically or food acceptable solvent, suspending agent or excipient for delivering the compound of formula (I), isomer, solvate, precursor, or pharmaceutically acceptable salt thereof of the present invention to animals or humans. The carrier may be a liquid or a solid.

It will be understood by those skilled in the art that, once the structure of the compounds of the present invention is known, the compounds of the present invention can be obtained by a variety of methods well known in the art, using well known starting materials, such as chemical synthesis or extraction from organisms (e.g., animals or plants), which are encompassed by the present invention.

In another aspect of the invention, a structural model of the interaction between Caspr1(203-355 amino acid peptide fragment) and IbeA (229-343 peptide fragment) is constructed for the first time, and the interaction amino acid hot spot residues are reported, wherein the hot spot residues in the IbeA (229-343 peptide fragment) are mainly concentrated on the α 1 and α helices, and form a small molecule binding pocket at the periphery, which is closely related to bacterial invasion.

In another aspect of the invention, IbeA (229-343 peptide fragment) is used as a target for the first time, and the obtained molecular model is virtually screened from a drug-like small molecular compound library to search for potential inhibitors from 213279 commercially available compounds. The first 50 small molecules with ordered binding energy were tested in vitro by Surface Plasmon Resonance (SPR) for their binding capacity titration to IbeA (229-343aa) purified protein, and 9 compounds showed direct binding capacity. Further, in vitro toxicity tests and bacterial invasion experiments prove that 4 molecules can obviously reduce or prevent the invasion of bacteria into hBMECs in a low cytotoxicity range.

The dosage form of the pharmaceutical composition of the present invention may be various, as long as it is a dosage form that enables the active ingredient to efficiently reach the body of a mammal. Such as may be selected from: a solution, suspension, tablet, capsule, powder, granule, or syrup. Depending on the type of disease to be treated by the compounds of the present invention, one skilled in the art may select a dosage form that is convenient to use.

The effective administration dose of the compound of formula (I) as an active ingredient may vary depending on the mode of administration and the severity of the disease to be treated. Or in a sustained release form, which may be adjusted to provide the optimum therapeutic response. For example, divided doses may be administered several times daily, or the doses may be proportionally reduced, or used in combination with other drugs having an effect on the treatment of E.coli meningitis, as is urgently required for the treatment condition.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, for which specific conditions are not noted in the following examples, are generally performed according to conventional conditions such as those described in J. SammBruk et al, molecular cloning protocols, third edition, scientific Press, 2002, or according to the manufacturer's recommendations.

EXAMPLE 1 Compound screening

Recently, neuron contact chaperonin 1 (contact-associated protein1, Caspr1) was identified in vivo in the laboratory of the applicant as a receptor of Escherichia coli virulence factor IbeA on BMECs, and the binding of the IbeA to the Laminin protein family domain consisting of Caspr1 extracellular segment 203-355aa through 229-343 amino acid fragment realizes the penetration of bacteria through the blood brain barrier. Based on the above research results, the three-dimensional spatial structures and interaction sites of Caspr1 (203-. The presence of a stable small molecule binding pocket in IbeA (229-.

In view of the fact that the Caspr1 and IbeA protein purification cannot meet the technical requirements of structure analysis, the three-dimensional space structures of Caspr1 (203-. Based on the prediction model, we found that IbeA (229-.

Then, the obtained molecular model is virtually screened from a drug-like small molecular compound library, and potential inhibitors are searched from 213279 commercial compounds. The method comprises the following specific steps:

1) preparing materials: the virtual screening software is Autodock Vina version 1.1.2 (version); the molecular three-dimensional structure display and processing software was PyMoL 4.4(DeLano Scientific, Palo Alto, CA); the compound database is from SPECS (http:// www.specs.net).

2) Preparing a database: considering the structural diversity and availability of sources of compounds, the SPECS database containing 200000 small molecules was chosen as the primary database in this study. The OpenBabel 2.4.0 software is used for carrying out preliminary screening on the small molecules in the database, and each compound can obtain a group of related data of physical and biological properties. The screened small molecules form a database A, and all the small molecules in the database A are used for subsequent molecular docking screening.

3) Preparation of protein active pocket: we selected the predicted small molecule binding pocket in IbeA (229-343) as the receptor structure for molecular docking. In the preparation process of the acceptor protein structure, water molecules in the acceptor structure are deleted, hydrogen atoms are added while nonpolar hydrogen is merged, and the electric field is set to gastiger. After the acceptor structure is prepared, molecular docking is initiated.

4) Molecular docking and screening: we used Autodock Vina 1.1.2 for molecular docking. Ready for work, all molecules in database a were docked into protein pockets, 9 docking conformations per small molecule, compounds were scored by scoring function in Vina, resulting in top 500 ranked compounds, and then considered better docking scores and consistent binding patterns (mainly with the mode of action of key amino acid residues CYS74, ASP278 and HIS280 in protein pockets). Thus, 50 compounds were screened for purchase and for subsequent bioactivity testing.

Table 1 shows the physicochemical properties and SPECS database numbers of the first 50-position small molecule compounds with high affinity to IbeA (229-343 amino acid peptide) obtained by screening based on IbeA hot spot amino acid residues and SPECS database.

FIG. 1 shows the hot spot amino acid residues that remain structurally stable in the Caspr1 and IbeA (229-343aa) complexes, A) showing that the hot spot amino acid residues in the Caspr1(203-355aa) -IbeA (229-343aa) complexes are displayed by calculation of binding energy by the MM/GBSA method on the Y axis, the X axis being the amino acid sequence, the key site sequence and the amino acid name being marked with an orange letter in the corresponding site, B) showing that the hot spot amino acid residue sites are marked in the Caspr-IbeA docking map, IbeA being shown by the green ribbon model, the key site amino acid name and sequence being marked with a black letter, Caspr1 being shown by the blue ribbon model, the key site amino acid name and sequence being marked with a red letter, and C) that the hot spot residues in IbeA are mainly concentrated on the α 1 and α 3 helices and form a small molecule binding pocket in the periphery, shown by the orange surface structure model, indicating that the IbeA (229-343aa) fragment is closely associated with bacterial attack.

FIG. 2 is a graph showing the affinity of the selected Top50 small molecule to the IbeA protein in the SPR in vitro test SPECS database. Of these, 9 small molecules showed direct binding to IbeA (229-343 aa).

FIG. 3 is a graph of HBMEC cytotoxicity of IbeA directly bound to small molecules, with half lethal concentration (IC50) of each compound shown by the orange dotted line, as measured by CCK-8 experiments, averaged in triplicate for each set of experiments. The results show that 5 small molecule compounds in total have low toxicity to HBMEC cells, i.e. the cell survival rate is still over 90% when the drug concentration reaches 100. mu.M (as shown in the figure).

Table 2 shows information of the bacteriostatic small-molecule lead compound small-molecule compounds obtained by screening based on IbeA hotspot amino acid residues and a SPECS database and verifying through in vitro experiments.

Example 2 Low toxicity test of Small molecule Compounds

The 50 small molecule compounds selected in example 1 were tested in the CCK-8 assay, and the tested cells were Human Brain Microvascular Endothelial Cells (HBMECs) derived from a source given by the professor Kim of John Hopkins medical school, USA. The HBMECs culture condition is that the RPMI 1640 culture medium is added with the following nutrients: 10% fetal bovine serum (FBS, Hyclone), 10% Nu-serum (BD Bosciences), 2mM glutamine, 1mM sodium pyrolate, 1 Xnon-essentia aminoacid, 1 XMEM vitamin. Incubator with constant temperature of 37 ℃ and 5% CO 2. Cytotoxicity at 2h/6h/24h/48h/72h was assessed by CCK-8 assay (Beyotime).

FIG. 3 is a graph of toxicity of small molecules that bind directly to IbeA on human brain microvascular endothelial cells (hBMEC) measured in the CCK-8 assay, as shown by the orange dotted line showing the median lethal concentration (IC50) of each compound, averaged over three replicates per group. The results show that 5 small molecule compounds in total have low toxicity to HBMEC cells, and are respectively: c₂₅H₂₂N₂O₃

(AG-205/33674026)、C₂₉H₂₂N₂O₈(AN-652/14563011)、C₂₈H₂₆Cl₂N₈(AK-968/11283147)、C₃₄H₂₂FN₃O₅S₂(AF-399/42017932) and C₂₉H₂₈Cl₂N₆O(AE-641/30154051)。

Example 3 Small molecule Compound invasion inhibition assay

1. Escherichia coli K1 strain was isolated clinically from cerebrospinal fluid of neonatal meningitis patients under conditions of 100ug/ml rifampicin in brain-heart infusion medium (BHI) at 37 ℃.

2. Small molecule powders purchased from SPECS, the Netherlands, were dissolved in DMSO to prepare a stock DMSO solution at 10.24mg/ml for use.

3. The culture conditions of human brain microvascular endothelial cells (hBMECs) are as follows: RPMI-1640 medium, containing 10% fetal bovine serum total bone serum (FBS, Hyclone), 10% Nu-serum (BD biosciences), 2mM glutamine, 1mM sodium sulfate, 1 Xnon-essential amino acid and 1 XMEM vitamin.37 deg.C, 5% CO2, 95% humidity conditions.

4. The hBMECs cells were seeded in 24-well plates and the culture medium was changed to experimental medium (RPMI-1640, 5% FBS, 1mol/L sodium pyruvate) as the cells grew into monolayers.

5. The small molecule to be detected (e.g., 50/100/150. mu.M, calculated for different molecular weights) is added at the desired concentration and incubated for 30 minutes. Each test group with different drug concentrations required a DMSO control group, added at the same volume as the drug solution.

6. To monolayer cultured hBMECs cells, equal amounts of E.coli were added and incubated at 37 deg.C, 5% CO2, 100% humidity for 1.5 hours, after which the experimental broth was discarded.

Washed 3 times with RPMI-1640.

7. After the extracellular bacteria were killed by addition of the experimental medium containing 100U/ml gentamicin, the culture was continued for another 1 hour.

8. The experimental culture medium containing gentamicin was discarded and washed 3 times with RPMI-1640.

9. The cells were lysed with 0.5% Triton X-100, and the cell lysate was spread on LB agar plates containing rifampicin, cultured overnight, and the number of colonies was counted.

The results of the invasion experiments are expressed in relative invasiveness (E44: drug addition/control group).

The experimental results prove that: small molecule C₂₉H₂₂N₂O₈(AN-652/14563011)，C₂₈H₂₆Cl₂N₈(AK-968/11283147)，C₃₄H₂₂FN₃O₅S₂(AF-399/42017932) and C₂₉H₂₈Cl₂N₆O (AE-641/30154051) has the effect of reducing or blocking bacterial invasion into the brain by competitively inhibiting the binding of the E.coli virulence factor IbeA protein to the host protein Caspr1 on brain endothelial cells.

Fig. 4 shows in vitro experiments to verify the effect of 5 low-toxicity small-molecule compounds on blocking the invasion of escherichia coli e. coli K1 into HBMEC, and the results show that 4 compounds can effectively block bacterial invasion within the low cytotoxicity range, which are: c₂₉H₂₂N₂O₈(AN-652/14563011)，C₂₈H₂₆Cl₂N₈(AK-968/11283147)，C₃₄H₂₂FN₃O₅S₂(AF-399/42017932) and C₂₉H₂₈Cl₂N₆O(AE-641/30154051)。

Example 4 comparison of Small molecule Compound invasion inhibition Effect

The best effect for blocking the bacterial invasion is small molecule C29H28Cl2N6O, the small molecule concentration can inhibit 90% + -6.0% of bacterial invasion when reaching 100 mu M, the blocking effect of C34H22FN3O5S2 and C29H22N2O8 molecules is the second, when the molecular concentration reaches 150 mu M, the bacterial invasion of 90% + -6.4% and 90% + -7.4% can be blocked respectively, the bacterial invasion of about 67.5% + -10.1% of bacteria can be inhibited when the molecular concentration reaches 150 mu M by C28H26Cl2N8 molecules, and the inhibiting effect at lower concentration is not obvious.

Example 5 construction of Butt-Joint Structure model of Small molecule Compound and I beA (229-343 amino acid peptide fragment)

The above-mentioned small molecule compound is butt-jointed with I beA (229-343 amino acid peptide segment), the structural mode diagram is shown in figure 5, wherein A is C₂₈H₂₆Cl₂N₈(AK-968/11283147); b is C₂₉H₂₈Cl₂N₆O (AE-641/30154051); c is C₃₄H₂₂FN₃O₅S₂(AF-399/42017932) D is C₂₉H₂₂N₂O₈(AN-652/14563011)。

The invention sets a foundation for preparing a target medicament for preventing/treating the bacterial meningitis of the newborn by adopting a new target treatment method from the aspects of intervening the interaction of a ligand (IbeA) -receptor (Caspr1) of a bacterium-host and closing pathogenic bacteria.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. Small molecule compound C₂₈H₂₆Cl₂N₈The application in preparing the medicine for treating the E.coli meningitis;

。

2. the small molecule compound C of claim 1₂₈H₂₆Cl₂N₈The application of the compound shown in the formula I or isomers, solvates or precursors thereof or pharmaceutically acceptable salts of the compound and the isomers, solvates or precursors in preparation of the medicine for treating the meningitis of the escherichia coli is characterized in that the compound shown in the formula I or the isomers, solvates or precursors thereof is combined in a targeted and competitive manner with the interaction hotspot amino acid residue of a receptor protein Caspr1 on the inner skin of a cerebral blood vessel in bacterial meningitis virulence factor IbeA protein, so that the combination of the two is blocked, and the aim of preventing the escherichia coli from passing through the blood brain barrier or blocking the meningitis escherichia coli from invading neurons is fulfilled.

3. The small molecule compound C of claim 2₂₈H₂₆Cl₂N₈The application of the IbeA protein in preparing the medicine for treating the Escherichia coli meningitis is characterized in that the IbeA protein is used as pathogenic bacteria of neonatal meningitisE.coliVirulence factors of K1.

4. The small molecule compound C of claim 2₂₈H₂₆Cl₂N₈The application of the IbeA-Caspr1 in preparing the medicine for treating the E.coli meningitis is characterized in that the interaction hotspot amino acid residues of IbeA-Caspr1 are R189, W196, R331, V332, N333, L337, S202, G325, I328, F329, N330, D336, R340, H342 and R344 are derived from Caspr1, V255, L319, F322, G323, T325, V326, G259, L261, T265, E266, M324, R329 and S331 are derived from IbeA.

5. The small molecule compound C according to any one of claims 1 to 4₂₈H₂₆Cl₂N₈The application of the medicine for treating the meningitis of the escherichia coli is characterized in that the medicine for treating the meningitis of the escherichia coli is used for blocking pathogenic bacteria to block meningitis escherichia coli from passing through a blood brain barrier or block meningitis escherichia coli from invading neurons.

6. The small molecule compound C according to any one of claims 1 to 5₂₈H₂₆Cl₂N₈The application of the medicine in preparing the medicine for treating the E.coli meningitis is characterized in that the medicine for treating the E.coli meningitis is a medicine for treating the central nervous system diseases which can easily pass through a blood brain barrier.

7. A pharmaceutical composition for treating e.coli meningitis, comprising: a compound of formula (I) or an isomer, solvate or precursor thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier;

formula I:

。

8. a pharmaceutical composition for treating e.coli meningitis, comprising: a compound of formula (I) or an isomer, solvate or precursor thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier; and other effective components for treating E.coli meningitis;

formula I:

。

9. the small molecule compound C of claim 8₂₈H₂₆Cl₂N₈The application of the traditional Chinese medicine composition in preparing a medicine for treating the E.coli meningitis is characterized in that other effective components for treating the E.coli meningitis are broad-spectrum antibiotics.

10. The small molecule compound C according to any one of claims 7-9₂₈H₂₆Cl₂N₈The application of the compound in preparing the medicine for treating the E.coli meningitis is characterized in that in the medicine composition, the pharmaceutically acceptable carrier is the compound, isomer, solvate, precursor or the compound of the formula IPharmaceutically or comestibly acceptable solvents, suspending agents or excipients for the delivery of a pharmaceutically acceptable salt to an animal or human; the carrier may be a liquid or a solid.