CN111518091B

CN111518091B - 1,2, 4-oxadiazole pyridine compound ahmu07 and application thereof as antibacterial drug

Info

Publication number: CN111518091B
Application number: CN202010321705.9A
Authority: CN
Inventors: 李增; 李明月; 滕霄; 郭亚辉; 王敏; 武明飞
Original assignee: Anhui Medical University
Current assignee: Anhui Medical University
Priority date: 2020-04-22
Filing date: 2020-04-22
Publication date: 2023-02-03
Anticipated expiration: 2040-04-22
Also published as: CN111518091A

Abstract

The invention discloses a 1,2, 4-oxadiazole pyridine compound ahmu07 and application thereof as an antibacterial drug, wherein the structural formula is shown as a formula I:

virtual screening and in-vitro antibacterial activity evaluation show that the 1,2, 4-oxadiazole pyridine compound ahmu07 has good antibacterial activity on MSSA and MRSA, and the MIC values are 1 mug/mL. The 1,2, 4-oxadiazole pyridine compound ahmu07 can be developed into a medicament for treating bacterial infection.

Description

1,2, 4-oxadiazole pyridine compound ahmu07 and application thereof as antibacterial drug

Technical Field

The invention belongs to the technical field of pharmacotherapeutics, and particularly relates to a 1,2, 4-oxadiazole pyridine compound ahmu07 and application thereof as an antibacterial drug.

Background

Bacterial resistance has become a global problem and the need for the development of new antibacterial agents has become very urgent. The main mode of obtaining the traditional antibacterial drugs is structural modification of the existing antibacterial drugs, but the development of the strategy becomes more and more difficult, so that the search of a new antibacterial drug action target and the development of the antibacterial drugs acting on the new target become more and more important ways. The filament Temperature-Sensitive protein Z (FtsZ) belongs to a bacterial division protein family, is widely distributed in prokaryotic cells, is highly conserved, has GTPase activity, is used as an initial regulation protein of bacterial division, and participates in the whole process of bacterial cell division. It has been found that inhibition of FtsZ function prevents the formation of bacterial cell divisions, which in turn leads to bacterial cell death.

Computer-Aided Drug Design (CADD) is an important method for rational Drug Design popular in recent years, and the basic principle of the CADD is a method for designing a novel Drug lead compound by simulating the interaction between a small-molecule Drug and a macromolecule. The development of the CADD technology greatly reduces the blindness and the contingency of new drug research and development, greatly shortens the cycle of new drug research and development, saves a large amount of manpower and financial resources, and becomes an important ring for new drug research and development. In the research, a CADD technology is utilized, a FtsZ crystal structure (PDB number: 4 DXD) from staphylococcus aureus is selected as a target spot, virtual screening is carried out, in-vitro anti-S.aureus activity evaluation is carried out on a screened compound, a partial screening result is analyzed by utilizing molecular dynamics simulation, and a FtsZ inhibitor is expected to be obtained, so that a certain reference is provided for researching and developing a new antibacterial drug.

Disclosure of Invention

The invention aims to make up for the defects of the prior art, and provides a 1,2, 4-oxadiazole pyridine compound ahmu07 and application thereof as an antibacterial drug by taking FtsZ protein as a virtual screening target, screening a lead compound with potential activity in a multiple screening mode and evaluating the in-vitro anti-S.aureus activity of the screened compound, and finds that the compound of the formula I has strong antibacterial activity.

In order to achieve the above object, the present invention provides the following technical solutions:

a1, 2, 4-oxadiazole pyridine compound ahmu07 has a structural formula shown in formula I:

the discovery method of the 1,2, 4-oxadiazole pyridine compound ahmu07 comprises the following steps:

(1) The small molecule compound library combined by the invention is used as an object, and the Discovery Studio 2017 software is adopted to virtually screen small molecule compounds which are targeted and combined with FtsZ and have potential druggability through multiple strategies.

(2) The antibacterial effect of small molecule compounds on s.

Another object of the present invention is to provide the use of the 1,2, 4-oxadiazolidine compound ahmu07 as described above in the preparation of antibacterial drugs which are pharmaceutically acceptable.

Further, the application medicine is any one of injection, tablets, pills, capsules, suspending agents or emulsion.

The invention has the advantages that:

the invention discovers that 1,2, 4-oxadiazole pyridine compound ahmu07 has good antibacterial activity on MSSA and MRSA through virtual screening and in-vitro antibacterial activity evaluation, and MIC values are 1 mug/mL. The 1,2, 4-oxadiazole pyridine compound ahmu07 can be developed into a medicament for treating bacterial infection.

Drawings

FIG. 1 shows ligand PC190723 Docking and result consistency evaluation in Docking modules Libdock, ligandfit, CDOCKER and Flexble Docking repeated 4DXD crystal complexes.

Detailed Description

The technical scheme of the invention is further explained by combining the specific examples as follows:

example 1

1. Docking software suitability evaluation

Four Docking modules, libdock, ligandfit, cdoccker and Flexble Docking, were used to dock the pro-ligand and FtsZ protein crystals and the results are shown in figure 1. The results show that the RMSD values of the ligand PC190723 after the four docking modules are docked and the RMSD value of the same ligand PC190723 in the FtsZ protein crystal compound before docking are all smaller than that of the ligand PC190723 in the FtsZ protein crystal compound before docking

Illustrate the four docking processesSequence and parameter settings are usually adapted to the study of molecular docking, and the docking results are reliable.

2. Virtual screening

The experimental steps are as follows: taking 1036 small molecular compounds in a small molecular library synthesized in the laboratory as screening objects, sequentially screening by sequentially using four Discovery Studio 2017 and four Docking modules Libdock, ligandfit, CDOCKER and Flexble Docking; subsequently, evaluation of the drug-like property was performed. The Libdock rigidly splices small molecule conformations into an active site pocket according to a hot zone matching principle, the LigandFit searches the flexible space conformation of molecules by adopting a Monte Carlo method based on the shape matching of a ligand and a receptor, and the two methods are used for rapid virtual screening; CDOCKER is based on a CHARMm force field, and each conformation is optimized at a receptor active site by adopting a simulated annealing method, so that the global energy minimum conformation is found, the CDOCKER docking result is more accurate, but the speed is lower, and the time consumption is about 5 times that of Libdock; the Flexible Docking simulates the states of a receptor and a ligand under physiological conditions, docking is carried out in a Flexible mode, and the receptor-ligand interaction process is accurately researched and used for accurate screening.

The experimental results are as follows: PC190723 was used as a control. The results show that 384 small molecule compounds are obtained after rapid screening by using Libdock and LigandFit modules; CDOCKER is used for optimizing each conformation of the 126 small molecule compounds obtained by screening at the active site of the receptor, and 32 small molecule compounds are obtained after accurate docking; carrying out full Flexible Docking on key amino acid residues by using a Flexible Docking module, and then, remaining 18 small molecule compounds; and finally, obtaining 11 small molecular compounds with patent medicine potential through evaluation of drug-like properties. See table 1.

TABLE 1Discovery Studio 2017 software four modules screening result score and in vitro anti-S.aureus activity evaluation

a:Libdock Score(Libdock)

b:Dock-Score(LigandFit)

c:-CDOCK_INTERACTION_ENERGY(CDOCKER)，

d:-CDOCK_INTERACTION_ENERGY(Flexible Docking)

MIC value (μ g/mL)

3. Evaluation of antibacterial Activity of Compound

The implementation steps are as follows: the antibacterial test was carried out by the method of the American Committee for clinical laboratory standards, and the antibacterial activity was examined by the broth dilution method at the final concentrations of 0.5. Mu.g/mL, 1. Mu.g/mL, 2. Mu.g/mL, 4. Mu.g/mL, 8. Mu.g/mL, 16. Mu.g/mL, 32. Mu.g/mL, 64. Mu.g/mL, 128. Mu.g/mL and 256. Mu.g/mL of the compound in Group 5. The method comprises the following specific operation steps: adding 100 mu L of nutrient broth culture medium into each well of a sterile 96-well plate, adding 95 mu L of nutrient broth culture medium into each well of the first row, respectively adding 5 mu L of a compound with the concentration of 10.24mg/mL into each well of the first row, uniformly mixing, performing half-time dilution, finally sucking 100 mu L of liquid from the 10 th well, finally adding 5 mu L of prepared bacterial liquid into each well, setting a blank control and a negative control, making a mark, incubating for 24h in an incubator at 37 ℃, observing the result, and calculating the MIC value.

The experimental results are as follows: as shown in Table 1, the selected compounds showed certain antibacterial activity against both MSSA and MRSA, and the MIC values ranged from 1-64. Mu.g/mL and 1-128. Mu.g/mL, respectively. Wherein the 1,2, 4-oxadiazole pyridine compound 12 shows the strongest activity, and MIC values are all 1 mug/mL.

4. Synthesis of Compound ahmu07

The intermediate is subjected to

(207mg, 0.5 mmol) was dissolved in 20mL of pyridine, followed by addition of 557mg,1.5mmol of 4-acetamidobenzenesulfonyl chloride), and the mixture was stirred at room temperature for 3 hours. The progress of the reaction was checked by TLC. After the reaction, dilute hydrochloric acid aqueous solution was added, and the mixture was placed on ice to precipitate a solid. Suction filtration is carried out, and the solid is dried. Purifying with dichloromethane in silica gel column chromatography: purification with methanol (300. ¹ H NMR(400MHz，DMSO)11.07(s，1H)，9.64(s，1H)， 8.34(d，J＝5.7Hz，1H)，8.03(d，J＝8.7Hz，2H)，7.85(ddd，J＝8.6，6.0， 2.6Hz，4H)，7.66(d，J＝8.5Hz，2H)，7.54(s，1H)，7.40(d，J＝8.8Hz，2H)， 7.38(dd，J＝15.0，5.7Hz，3H)，3.03(s，3H).MS(ESI)：m/z 594.6[M+H] ⁺ 。

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. A1, 2, 4-oxadiazole pyridine compound ahmu07 is characterized in that the structural formula is shown as a formula I:

formula I.

2. Use of the 1,2, 4-oxadiazolopyridine compound ahmu07 according to claim 1 in the manufacture of a pharmaceutically acceptable antibacterial medicament.

3. The use according to claim 2, wherein the medicament for use is any one of an injection, a tablet, a pill, a capsule, a suspension or an emulsion.