CN108231206A - A kind of screening technique and lead compound of moulting hormone analog - Google Patents

A kind of screening technique and lead compound of moulting hormone analog Download PDF

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CN108231206A
CN108231206A CN201810010234.2A CN201810010234A CN108231206A CN 108231206 A CN108231206 A CN 108231206A CN 201810010234 A CN201810010234 A CN 201810010234A CN 108231206 A CN108231206 A CN 108231206A
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receptor
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张莉
胡雪萍
胡松
董雅雯
王家哲
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China Agricultural University
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    • G16INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
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Abstract

The present invention provides a kind of screening technique of moulting hormone analog, including:S1 active pocket) is determined;S2) according to receptors ligand Pharmacophore Model and ligand Pharmacophore Model screening SPECS libraries, database 1 is established;S3) compound in database 1 is clustered and quasi-medicated property is analyzed, establishes database 2;S4) compound in database 2 with receptor is docked and establishes database 3, screens its receptor-binding activity, obtains the compound of virtual screening;S5) combination of compound and receptor is active in garbled data library 2, obtains the compound of virtual screening;S6 the compound of virtual screening) is subjected to ecdysone receptor combination determination of activity, the lead compound hit;S7) lead compound based on the hit carries out similarity retrieval.Virtual screening method provided by the invention can reduce the quantity of experiment screening compound, shorten the R&D cycle, reduce financial cost.

Description

A kind of screening technique and lead compound of moulting hormone analog
Technical field
The invention belongs to technical field of pharmaceuticals more particularly to a kind of screening techniques of moulting hormone analog and guide's chemical combination Object.
Background technology
Ecdysone receptor is by husking receptor protein (Ecdysteroid Receptor, EcR) and super valve albumen The dynamic complex of (ultraspiracle, USP) composition.Both albumen are nuclear receptor superfamily (nuclear receptor Super-family) forming member.Ecdysone receptor EcR/USP is existed only in insect bodies, it can with moulting hormone and Moulting hormone analog carries out single-minded combination and forms compound, and then is combined with target gene promoter element, and regulation and control insect sloughs off Skin.This regulation process influences insect normal growth and development and breeding, therefore ecdysone receptor is a kind of important pesticide Action target, moulting hormone analog can be as the potential drugs of pest control.
At present, the steroid moulting hormone analog obtained is extracted more than 500 kinds from natural animal source or plant source, still Due to complicated, synthesis difficulty is high, and stability is poor and the problem of to insect cuticle poor permeability, leads to such compound So far it is not all actually used in field of pest control.1988, Rohm and Haas Company (US) Independenec Mall West, Philadelphia, Pennsy Lvania 1 of the U.S. was to a large amount of natural or artificial conjunction On the basis of being screened into compound, developing first different from natural ecdysone structure, similary there is husking to swash The bishydrazide insect growth regulator, IGR RH-5849 (suppression food hydrazine) of plain activity.Has the medicament of 5 commercializations at present:Tebufenozide (RH-5992), methoxyfenozide (RH-2485), chlorine tebufenozide (RH-0345), ring tebufenozide (ANS-118), furan tebufenozide (JS-118).But due to being used for a long time, have there are resistance problems in bishydrazide compounds.
In recent years, some moulting hormone analogs, such as γ-methyl-γ-lactams, oxazoline, benzamides chemical combination Object, Tetrahydroquinolinesas, acetylamino ketone compounds, 8-O- acetyl Harpagides, maocrystal E etc. are sent out successively It is existing, but because the more low reason of live body activity is not all developed further.Therefore, it is badly in need of screening and finds that a kind of skeleton is novel, easily In synthesis and the smaller high activity moulting hormone analog of financial cost.
The conventional method of drug screening is random screening, i.e., is largely sieved according to pharmacological model from a large amount of compound Choosing, this method heavy workload is time-consuming longer, costly, and blindness is bigger.With Computer-Aided Drug Design The appearance of method, computer virtual screening provide a kind of quick, economical and effective drug screening method.The technology is by grinding Study carefully the three-dimensional structure of particular target macromolecular, establish effective Pharmacophore Model, by technological means such as molecular dockings, in business Change in small molecule libraries, filtering out has interaction with target macromolecule or meets the micromolecular compound of Pharmacophore Model, from And shorten the medicament research and development period, and increase drug screening probalility of success, it is cost-effective.Therefore, a kind of quick screening husking is established to swash The method of plain analog is of great significance.
Invention content
In view of this, the technical problem to be solved in the present invention is to provide a kind of screening moulting hormone rapidly and efficiently similar The method and lead compound of object.
The present invention provides a kind of screening technique of moulting hormone analog, including:
S1 the three-dimensional structure of EcR/USP dimers and ligand eutectic compound) is obtained from Protein Data Bank, Analysis is compared, determines active pocket;
S2 receptor-ligand Pharmacophore Model and ligand Pharmacophore Model) are built according to active pocket, and according to receptor-match Body Pharmacophore Model establishes database 1 with ligand Pharmacophore Model screening SPECS libraries, qualified compound;
S3) compound in the database 1 is clustered and quasi-medicated property is analyzed, qualified compound is selected to build Vertical database 2;
S4) using molecular docking programs, the compound in the database 2 with receptor is docked, database 3 is established, adopts With the receptor-binding activity of compound in QSAR model prediction garbled datas library 3, the compound of first virtual screening is obtained;
S5) using the combination activity of compound and receptor in MM-PBSA methods prediction garbled data library 2, second batch is obtained The compound of virtual screening;
S6) by the compound of the compound of first virtual screening and second batch virtual screening carry out moulting hormone by Body combination determination of activity, the lead compound hit;
S7 the lead compound) based on the hit carries out similarity retrieval to the compound in the SPECS libraries, obtains The compound of third batch virtual screening;
Wherein, the step S4) and step S5) have no point of sequencing.
Preferably, the step S1) in compare analysis using Align/Superpose modules in MOE;The EcR/USP Dimer and ligand eutectic compound be selected from Lepidoptera tobacco budworm EcR/USP dimers and ponasterone eutectic compound, Lepidoptera tobacco budworm EcR/USP dimers and bishydrazide BYI06830 eutectic compounds, Homoptera Bemisia tabaci EcR/USP bis- Aggressiveness and ponasterone eutectic compound;Elytrum hot eyes model one's literary or artistic style on that of the ancients steal EcR/USP dimers and ponasterone eutectic compound, Lepidoptera tobacco budworm EcR/USP dimers and 20- hydroxyecdysones eutectic compound, Lepidoptera bollworm EcR/USP dimerization Body and bishydrazide BYI08346 eutectic compounds.
Preferably, the step S2) in structure receptor-ligand Pharmacophore Model and the method for ligand Pharmacophore Model be: Using Ligand interaction module analysis extracting EcR/USP dimers and natural ecdysone in ligand eutectic compound With the common pharmacodynamic properties of bishydrazide compounds, common pharmacodynamic properties are then based on using Pharmacophore module constructions Receptor-ligand Pharmacophore Model and ligand Pharmacophore Model;The common pharmacodynamic properties include hydrogen bond receptor, hydrogen bond donor, row Except volume and hydrophobic effect.
Preferably, the step S4) in molecular docking using Dock modules carry out, laying method Triangle Matcher;After molecular compound is docked with receptor, using Affinity dG, Alpha HB, ASE scoring functions combination evaluation point The binding ability of sub- compound and receptor.
Preferably, the QSAR models are established in accordance with the following methods:Choose bishydrazide compounds composing training collection, data Compound in library 3 forms test set, and meter is iterated using the AutoQSAR modules in MOE to the compound in test set It calculates, obtains QSAR models.
Preferably, the step S5) it is middle using the knot of compound and receptor in MM-PBSA methods prediction garbled data library 2 Conjugated free energy of the activity based on Amber12 is closed to calculate.
The present invention also provides a kind of screening technique of moulting hormone analog, including:
A1 the three-dimensional structure of EcR/USP dimers and ligand eutectic compound) is obtained from Protein Data Bank, Analysis is compared, determines active pocket;
A2) molecular compound in SPECS libraries is clustered and quasi-medicated property is analyzed, qualified compound is selected to build Vertical database 1 ';
A3 receptor-ligand Pharmacophore Model and ligand Pharmacophore Model) are built according to active pocket, and according to receptor-match Body Pharmacophore Model and ligand Pharmacophore Model garbled data library 1 ', qualified compound establishes database 2 ';
A4) using molecular docking programs, the compound in the database 2 ' with receptor is docked, establishes database 3 ', Using the receptor-binding activity of 3 ' middle compound of QSAR model prediction garbled datas library, the chemical combination of first virtual screening is obtained Object;
A5) using 2 ' middle compound of MM-PBSA methods prediction garbled data library and the combination activity of receptor, second batch is obtained The compound of virtual screening;
A6) by the compound of the compound of first virtual screening and second batch virtual screening carry out moulting hormone by Body combination determination of activity, the lead compound hit;
A7 the lead compound) based on the hit carries out similarity retrieval to the compound in the SPECS libraries, obtains The compound of third batch virtual screening;;
Wherein, the step A1) and step A2) have no point of sequencing;The step A4) and step A5) have no elder generation Point of sequence afterwards.
The present invention also provides the lead compound for mosquito larvae with killing activity that above-mentioned screening technique obtains, tools There are VSI-1, VSI-2, VSI-3, VSI-4, VSI-5, VSI-6, VSI-7, VSI-8, VSI-10, VSI-12, VSI-15, VSI- 16th, structure shown in VSI-19, VSI-20, VSI-23, VSIII-4, VSIII-39, VSIII-50:
What is obtained the present invention also provides above-mentioned screening technique has the guide's chemical combination for combining activity with ecdysone receptor Object, have VSI-3, VSI-9, VSI-10, VSI-14, VSI-15, VSI-16, VSI-17, VSI-18, VSI-20, VSII-2, VSII-7、VSII-17、VSIII-1、VSIII-8、VSIII-9、VSIII-12、VSIII-15、VSIII-26、VSIII-28、 Structure shown in VSIII-36, VSIII-37, VSIII-38, VSIII-43, VSIII-63, VSIII-64:
The present invention also provides above-mentioned screening technique obtain to diamondback moth have ecdysone receptor combine activity and/or The moulting hormone analog of live body killing activity, compound or its pharmacy with structure shown in formula one, formula two, formula three, formula four Upper acceptable salt, ester, solvate:
Wherein, R1With R1' it is each independently selected from substituted or unsubstituted cycloalkyl, substituted or unsubstituted Heterocyclylalkyl, substituted or unsubstituted heterocycloalkenyl, is substituted or unsubstituted substituted or unsubstituted cycloalkenyl group Fused ring aryl, substituted or unsubstituted condensed hetero ring base, substituted or unsubstituted aryl or substituted or do not taken The heteroaryl in generation;
R2With R2' it is each independently selected from substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycle alkane Base, substituted or unsubstituted cycloalkenyl group, substituted or unsubstituted heterocycloalkenyl, substituted or unsubstituted condensed ring Aryl, substituted or unsubstituted condensed hetero ring base, substituted or unsubstituted aryl or substituted or unsubstituted miscellaneous Aryl;
R3With R3' be each independently selected from H, halogen, substituted or unsubstituted formamido, be substituted or do not taken The formic acid esters in generation, substituted or unsubstituted amino, substituted or unsubstituted nitro, cyano, acetonitrile-base, hydroxyl, hydrogen Sulfenyl, substituted or unsubstituted alkyl, substituted or unsubstituted miscellaneous alkyl;
R4With R4' be each independently selected from H, halogen, substituted or unsubstituted formamido, be substituted or do not taken The formic acid esters in generation, substituted or unsubstituted amino, substituted or unsubstituted nitro, cyano, acetonitrile-base, hydroxyl, hydrogen Sulfenyl, substituted or unsubstituted alkyl, substituted or unsubstituted miscellaneous alkyl;
R5With R5' it is each independently selected from H, halogen, substituted or unsubstituted amino, substituted or unsubstituted Nitro, cyano, acetonitrile-base, hydroxyl, sulfhydryl, substituted or unsubstituted alkyl, substituted or unsubstituted miscellaneous alkyl, Substituted or unsubstituted cycloalkyl, substituted or unsubstituted Heterocyclylalkyl, substituted or unsubstituted cycloalkenyl group, It is substituted or unsubstituted heterocycloalkenyl, substituted or unsubstituted fused ring aryl, substituted or unsubstituted thick miscellaneous Ring group, substituted or unsubstituted aryl, substituted or unsubstituted urea groups, substituted or unsubstituted phenoxy group, Or substituted or unsubstituted heteroaryl, substituted or unsubstituted ester group.
The present invention provides a kind of screening technique of moulting hormone analog, including:S1) from Protein Data Bank The middle three-dimensional structure for obtaining EcR/USP dimers and ligand eutectic compound, compares analysis, determines active pocket;S2) according to work Property pocket structure receptor-ligand Pharmacophore Model and ligand Pharmacophore Model, and according to receptor-ligand Pharmacophore Model and ligand Pharmacophore Model screening SPECS libraries, qualified molecular compound establish database 1;S3) to point in the database 1 Sub- compound is clustered and quasi-medicated property analysis, and qualified compound is selected to establish database 2;S4 molecular docking) is utilized Molecular compound in the database 2 with receptor is docked, establishes database 3 by program, and number is screened using QSAR model predictions According to the receptor-binding activity of compound in library 3, the compound of first virtual screening is obtained;S5 it) is predicted using MM-PBSA methods The combination activity of compound and receptor, obtains the compound of second batch virtual screening in garbled data library 2;S6) by described first The compound and the compound of second batch virtual screening for criticizing virtual screening carry out ecdysone receptor combination determination of activity, are ordered In lead compound;S7 the lead compound) based on the hit carries out similitude inspection to the compound in the SPECS libraries Rope obtains the compound of third batch virtual screening;Wherein, the step S4) and step S5) have no point of sequencing.With showing There is technology to compare, virtual screening method provided by the invention can reduce the quantity of experiment screening compound, shorten the R&D cycle, drop Low financial cost.
Description of the drawings
Fig. 1 is the overlapping figure of 6 eutectic compounds;
Fig. 2 is receptor-ligand Pharmacophore Model figure;
Fig. 3 is ligand Pharmacophore Model figure;
Fig. 4 is the enrichment curve graph of five kinds of scoring functions and combinations thereof;
Fig. 5 is binding activity result figure of the compound to diamondback moth ecdysone receptor of two batches virtual screening;
Fig. 6 is binding activity result figure of the compound to diamondback moth ecdysone receptor of third batch virtual screening.
Specific embodiment
Below in conjunction with the embodiment of the present invention, the technical solution in the embodiment of the present invention is clearly and completely described, Obviously, described embodiment is only part of the embodiment of the present invention, instead of all the embodiments.Based in the present invention Embodiment, those of ordinary skill in the art's all other embodiments obtained without making creative work, all Belong to the scope of protection of the invention.
The present invention provides a kind of screening technique of moulting hormone analog, including:
S1 the three-dimensional structure of EcR/USP dimers and ligand eutectic compound) is obtained from Protein Data Bank, Analysis is compared, determines active pocket;
S2 receptor-ligand Pharmacophore Model and ligand Pharmacophore Model) are built according to active pocket, and according to receptor-match Body Pharmacophore Model establishes database 1 with ligand Pharmacophore Model screening SPECS libraries, qualified molecular compound;
S3) molecular compound in the database 1 is clustered and quasi-medicated property is analyzed, selects qualified chemical combination Object establishes database 2;
S4) using molecular docking programs, the molecular compound in the database 2 with receptor is docked, establishes database 3, using the receptor-binding activity of compound in QSAR model prediction garbled datas library 3, obtain the chemical combination of first virtual screening Object;
S5) using the combination activity of compound and receptor in MM-PBSA methods prediction garbled data library 2, second batch is obtained The compound of virtual screening;
S6) by the compound of the compound of first virtual screening and second batch virtual screening carry out moulting hormone by Body combination determination of activity, the lead compound hit;
S7 the lead compound) based on the hit carries out similarity retrieval to the compound in the SPECS libraries, obtains The compound of third batch virtual screening;
Wherein, the step S4) and step S5) have no point of sequencing.
The three-dimensional structure of EcR/USP dimers and ligand eutectic compound is obtained from Protein Data Bank, is compared Analysis, determines active pocket:Wherein, the EcR/USP dimers and ligand eutectic compound is known to those skilled in the art EcR/USP dimers and ligand eutectic compound, have no special limitation, preferably Lepidoptera cigarette bud night in the present invention Moth EcR/USP dimers and ponasterone eutectic compound (PDB:1R1K), Lepidoptera tobacco budworm EcR/USP dimers with Bishydrazide BYI06830 eutectic compounds (PDB:1R20), Homoptera Bemisia tabaci EcR/USP dimers and ponasterone are total to Brilliant compound (PDB:1Z5X);Elytrum hot eyes, which is modeled one's literary or artistic style on that of the ancients, steals EcR/USP dimers and ponasterone eutectic compound (PDB: 2NXX), Lepidoptera tobacco budworm EcR/USP dimers and 20- hydroxyecdysones eutectic compound (20E) (PDB:2R40), squama Wing mesh bollworm EcR/USP dimers and bishydrazide BYI08346 eutectic compounds (PDB:3IXP);The comparison analysis is adopted With Align/Superpose modules in MOE, Align/Superpose modules in MOE 2014.08 are more preferably used;Through comparing Analysis, for determining active pocket as shown in FIG. 1, FIG. 1 is the overlapping figure of 6 eutectic compounds, the ligand in wherein 1R1K is green Color, 1R20 are blue, and 1Z5X is cyan, and 2NXX is brown, and 2R40 is purple, and 3IXP is grey;Colored region is ligand binding Pocket.
Receptor-ligand Pharmacophore Model and ligand Pharmacophore Model are built according to active pocket, the method for the structure is excellent It is selected as:EcR/USP dimers are extracted with naturally being sloughed off in ligand eutectic compound using Ligand interaction module analysis The common pharmacodynamic properties of skin hormone and bishydrazide compounds are then based on common pharmacodynamic properties using Pharmacophore moulds Block builds receptor-ligand Pharmacophore Model and ligand Pharmacophore Model;The common pharmacodynamic properties include hydrogen bond receptor, hydrogen bond supplies Body, excluded volume and hydrophobic effect;Ligand interaction modules are preferably the Ligand interaction moulds in MOE Ligand interaction modules in block, more preferably MOE 2014.08;The Pharmacophore modules are preferably Pharmacophore modules in Pharmacophore modules in MOE, more preferably MOE 2014.08.
According to the present invention, receptor-ligand Pharmacophore Model and ligand Pharmacophore Model are more preferably built in accordance with the following methods:
Using ligand in 6 eutectic compounds of Ligand interaction module analysis in software MOE 2014.08 Interaction between receptor extracts the common pharmacodynamic properties of natural ecdysone and bishydrazide compounds.4 naturally slough off All there are Thr343, Asn504, Ile339, Trp526 in the binding cavity of skin hormone, wherein Thr343, Asn504 does hydrogen bond confession Body, Ile339, Trp526 form hydrophobic region around the alkyl chain of natural ecdysone, and there are important hydrophobic effects.2 All there are Thr343, Tyr408, Ile339, Trp526, Met380 in the binding cavity of bishydrazide compounds, wherein Thr343 is done Hydrogen bond donor, Tyr408 do hydrogen bond receptor, and Ile339, Trp526, Met380 are respectively around the aromatic ring of bishydrazide, tertiary butyl Form two hydrophobic regions.Using Pharmacophore modules in software MOE2014.08, based on Ligand interaction The analysis result of module builds two Pharmacophore Models:Receptor-ligand Pharmacophore Model (as shown in Figure 2), ligand pharmacophore mould Type (as shown in Figure 3).
To verify the separating capacity of Pharmacophore Model high low activity to compound, by 25 high activity bishydrazide chemical combination Object, 14 low activity bishydrazide compounds and 6 natural ecdysones are as test set.In receptor-ligand Pharmacophore Model, A hydrogen bond donor site is defined at Thr343, a hydrogen bond receptor site is defined at Tyr408, Asn504 defines a hydrogen Key donor site defines a hydrophobic effect at alkyl chain, tertiary butyl.The positive accuracy of receptor-ligand Pharmacophore Model= 100%, negative accuracy=50%.In ligand Pharmacophore Model, Thr343 corresponds at 17-C=O, 15-N on ligand Define one be hydrogen bond receptor site, Tyr408 correspond to ligand on 14-N place be hydrogen bond donor site, Ile339, Trp526, Met380 corresponds to the aromatic ring of ligand and alkyl is two hydrophobic effect sites.And excluded volume is added to binding pocket atom, Optimize pharmacophore, excluded volume radius R=1.72.Positive accuracy=100% of ligand Pharmacophore Model, negative accuracy= 78%.
According to receptor-ligand Pharmacophore Model and ligand Pharmacophore Model screening SPECS libraries, qualified molecule chemical combination Object establishes database 1, is particularly preferred as:Screening marking is carried out to SPECS libraries using two Pharmacophore Models of structure, will be given a mark Value S values are ranked up, and S values are more negative, show that the matching degree of micromolecular compound and pharmacophore is higher, select S values forward 8851 compounds, establish database 1.
Molecular compound in the database 1 is clustered and quasi-medicated property is analyzed, selects qualified compound Establish database 2;The cluster and the method for quasi-medicated property analysis are method well known to those skilled in the art, and it is special to have no Limitation, present invention preferably employs the Calculate Descriptors modules in MOE, more preferably using in MOE 2014.08 Calculate Descriptors modules;It is more specific to be preferably:Using the Calculate in software MOE2014.08 Descriptors modules, cluster database 1 and quasi-medicated property is analyzed.Calculate the weight of all compounds in database 1 (molecular weight)、ClogP(log octanol/water partition coefficient)、HBA(number of H-bond acceptor atoms)、HBD(number of H-bond donor atoms)、ROB(number of Rotatable bonds) and ARB (number of aromatic atoms) parameter, according to class pesticide rule, selector Conjunction condition:weight<435、ClogP<6、HBA<6、HBD<2、ROB<9、ARB<17 micromolecular compound establishes database 2, Totally 2538 compounds.
Using molecular docking programs, the molecular compound in the database 2 with receptor is docked, establishes database 3;Institute Molecular docking programs are stated as molecular docking programs well known to those skilled in the art, have no special limitation, the present invention is excellent Choosing more preferably selects the compound in database 2 using Dock modules in MOE 2014.08 silent using Dock modules in MOE The docking agreement recognized carries out molecular docking;Laying method is Triangle Matcher;The receptor is those skilled in the art Well known receptor, has no special limitation, preferably using 1R20 as receptor in the present invention;After docking, it is preferred to use marking letter Number carries out pose rescoring, and database 3 is established according to sequence;The scoring functions are beaten to be well known to those skilled in the art Divide function, special limitation is had no, after preferably being docked in the present invention using molecular compound with receptor, using Affinity DG, Alpha HB, ASE scoring functions combination evaluation molecular compounds and receptor binding ability.According to according to Wang Ren little et al. Research to scoring functions, a variety of scoring functions combinations can improve the reliability of molecular docking marking.To select accumulation ability High scoring functions, by 6 compounds (tebufenozide, furan tebufenozide, methoxyfenozide, chlorine tebufenozide, the ring worm of commercialization Hydrazides, suppression food hydrazine) and known effective 4 compounds to ecdysone receptor (BYI08346, BYI09181, BYI08738, BYI06934), it is added to and picks out 1000 compounds from SPECS libraries at random, forms accumulation ability evaluation library.First to this 1010 small molecules carry out molecular docking by receptor of 1R20.After molecular docking, docked using the pose rescoring in MOE Agreement is to docking conformation S_A:GBVI/WSAdG、S_B:Affinity dG、S_C:Alpha HB、S_D:ASE、S_E: Five kinds of scoring functions of London dG are given a mark again, then carry out accumulation ability to different scoring functions and combination of function Evaluation, obtains the enrichment curve graph of five kinds of scoring functions and combinations thereof, as shown in Figure 4.It is found through experiments that, Affinity dG+ The combination accumulation ability of tri- functions of Alpha HB+ASE is most strong, therefore the molecular docking part in virtual screening, selects first Compound is docked with the docking agreement of acquiescence, is then combined with Affinity dG, Alpha HB, ASE scoring functions Pose rescoring are carried out, database 3 is established according to sequence;30%~35% compound is built before the preferred selection of sequence Vertical database 3,32% compound establishes database 3 before more preferably choosing, totally 790 compounds.
Using the receptor-binding activity of compound in QSAR model prediction garbled datas library 3, first virtual screening is obtained Compound.The receptor-binding activity using QSAR model prediction screening compounds is preferably based on QuaSAR modules in MOE, QuaSAR modules in more preferably based on MOE 2014.08;The QSAR models preferably pass through the double hydrazine acyl class chemical combination of quantitative analysis The structure-activity relation of object is established, and is more preferably established according to following steps:Choose bishydrazide compounds composing training collection, number Test set is formed according to the compound in library 3, meter is iterated using the AutoQSAR modules in MOE to the compound in test set It calculates, obtains QSAR models;The number of compound is preferably 81~101 in the training set, more preferably 91;The test The number for concentrating compound is preferably 31~51, more preferably 41;According to the present invention, this step is more preferably specially:Root According to structure diversity, 91 bishydrazide compounds composing training collection are selected, 41 compounds form test set, to test set Object is closed using the AutoQSAR modules in MOE, iterative calculation selects suitable descriptor combinations, finally obtaining model eq (1) is QSAR models.
Observations:91Descriptors:8 R2:0.64066 XR2:0.56736
Eq (1) model analysis:ASA-(negative accessible surface area)、E_tor(torsion energy)、PEOE_VSA+1(total positive 1 vdw surface area)、PEOE_VSA_NEG(total Negative vdw surface area), SlogP_VSA4 (4 slogp of bin (0.10,0.15)), a_hyd (number of hydrophobic atoms)、b_count(number ofbonds)、vsurf_CW2(capacity factor at-0.5)。
Model eq (1) is 36/41=87.8% to the accuracy rate that test set compound activity is predicted, utilizes eq (1) logarithm Activity Prediction is carried out according to library 3, obtains 23 candidate compounds (VSI-1~VSI-23) of first virtual screening.
Using the combination activity of compound and receptor in MM-PBSA methods prediction garbled data library 2, it is virtual to obtain second batch The compound of screening;In the research of early period, MM-PBSA can preferably predict natural ecdyson and bishydrazide compounds And the combination activity of receptor.Wherein, the preferred base of combination activity that screening compounds and receptor are predicted using MM-PBSA methods It is calculated in the Conjugated free energy of Amber12;The prediction screening preferably follows the steps below:By the chemical combination in database 2 Object is rigidly docked in the binding cavity of receptor, then carries out 20ps Dynamics Optimizations to it, using MM-PBSA methods predict with by The combination activity of body;The receptor is receptor well known to those skilled in the art, has no special limitation, excellent in of the invention It is selected as 1R20.
The compound of first virtual screening and the compound of second batch virtual screening are subjected to ecdysone receptor With reference to determination of activity, the lead compound hit;The method for measuring is that method well known to those skilled in the art is Can, special limitation is had no, preferably:With the pure protein (EcRB, USP) of in-vitro transcription translation for receptor, with the low of pH=7.9 Salt buffer is buffer solution, and setting competitive binding group, summary are combined, non-specific binding group, and every group sets three repetitions;Competition knot It is combined:The diamondback moth ecdysone receptor working solution (EcR+USP) of 8 μ l, 4.8 μ l [3H] PonA working solution (final concentrations 15nM), the 3.2 certain density untested compounds of μ l;Summary is combined (TB):The diamondback moth ecdysone receptor working solution of 8 μ l (EcR+USP), 4.8 μ l [3H] PonA working solutions (final concentration 15nM), 3.2 μ l less salts buffer supplement volumes to 16 μ l;It is non- Specific binding group:The diamondback moth ecdysone receptor working solution (EcR+USP) of 8 μ l, 4.8 μ l [3H] PonA working solutions are (eventually Concentration 15nM), the PonA, 16 μ l of total volume of 3.2 μ l not tape labels.Above-mentioned reaction solution 25 DEG C of reaction 90min in water-bath, instead It should finish and be placed on ice.Cylinder type filter (diameter 25mm, effective diameter 21mm) vacuum filtration separation, filter membrane select nitric acid fine NC45 is tieed up, the ice-cold elution buffer filter washes film of 3ml 3 times adds 3ml scintillation solutions, and liquid scintillation counter obtains data.Percentage bound It is calculated as follows, experimental result is as shown in figure 5, the compound that Fig. 5 is two batches virtual screening is casted off a skin to diamondback moth and swashed The binding activity result figure of plain receptor.The compound high for combining activity, gradient reduce concentration, measure IC50
Lead compound based on the hit carries out similarity retrieval to the compound in the SPECS libraries, obtains the The compound of three batches of virtual screenings.
According to the present invention, ecdysone receptor preferably is carried out to the compound of the third batch virtual screening and combines activity survey Examination, the method for the test is same as above, and details are not described herein, and the compound for obtaining third batch virtual screening casts off a skin to diamondback moth The binding activity result figure of hormone receptor is as shown in Figure 6.
According to the present invention, preferably also to the compound of the compound of first virtual screening, second batch virtual screening and the The compound of three batches of virtual screenings carries out receptor radioligand analysis, measures the combination activity of itself and ecdysone receptor.
The present invention also provides the screening technique of another moulting hormone analog, including:
A1 the three-dimensional structure of EcR/USP dimers and ligand eutectic compound) is obtained from Protein Data Bank, Analysis is compared, determines active pocket;
A2) molecular compound in SPECS libraries is clustered and quasi-medicated property is analyzed, qualified compound is selected to build Vertical database 1 ';
A3 receptor-ligand Pharmacophore Model and ligand Pharmacophore Model) are built according to active pocket, and according to receptor-match Body Pharmacophore Model and ligand Pharmacophore Model garbled data library 1 ', qualified compound establishes database 2 ';
A4) using molecular docking programs, the compound in the database 2 ' with receptor is docked, establishes database 3 ', Using the receptor-binding activity of 3 ' middle compound of QSAR model prediction garbled datas library, the chemical combination of first virtual screening is obtained Object;;
A5) using 2 ' middle compound of MM-PBSA methods prediction garbled data library and the combination activity of receptor, second batch is obtained The compound of virtual screening;
A6) by the compound of the compound of first virtual screening and second batch virtual screening carry out moulting hormone by Body combination determination of activity, the lead compound hit;
A7 the lead compound) based on the hit carries out similarity retrieval to the compound in the SPECS libraries, obtains The compound of third batch virtual screening;;
Wherein, the step A1) and step A2) have no point of sequencing;The step A4) and step A5) have no elder generation Point of sequence afterwards.
I.e. cluster and quasi-medicated property analysis can be placed in screens it according to receptor-ligand Pharmacophore Model with ligand Pharmacophore Model Preceding progress, instructs the screening of lead compound, other steps are same as mentioned above, and details are not described herein.
The present invention also provides guide's chemical combination for mosquito larvae with killing activity that a kind of above-mentioned screening technique obtains Object, have VSI-1, VSI-2, VSI-3, VSI-4, VSI-5, VSI-6, VSI-7, VSI-8, VSI-10, VSI-12, VSI-15, Structure shown in VSI-16, VSI-19, VSI-20, VSI-23, VSIII-4, VSIII-39, VSIII-50:
What is obtained the present invention also provides a kind of above-mentioned screening technique has the guide for combining activity with ecdysone receptor Compound, preferably with diamondback moth ecdysone receptor have combine activity lead compound, have VSI-3, VSI-9, VSI-10、VSI-14、VSI-15、VSI-16、VSI-17、VSI-18、VSI-20、VSII-2、VSII-7、VSII-17、VSIII- 1、VSIII-8、VSIII-9、VSIII-12、VSIII-15、VSIII-26、VSIII-28、VSIII-36、VSIII-37、 Structure shown in VSIII-38, VSIII-43, VSIII-63, VSIII-64:
The present invention also provides what a kind of above-mentioned screening technique obtained there is ecdysone receptor to combine activity diamondback moth And/or the moulting hormone analog of live body killing activity, have formula one, formula two, formula three, the compound of structure shown in formula four or its Pharmaceutically acceptable salt, ester, solvate:
Wherein, R1With R1' it is each independently substituted or unsubstituted cycloalkyl, substituted or unsubstituted miscellaneous It is cycloalkyl, substituted or unsubstituted cycloalkenyl group, substituted or unsubstituted heterocycloalkenyl, substituted or unsubstituted Fused ring aryl, substituted or unsubstituted condensed hetero ring base, substituted or unsubstituted aryl are substituted or unsubstituted Heteroaryl;Above-mentioned substituent group is preferably one or more in the alkyl and halogen of C1~C5, more preferably H, C1~C3 Alkyl and halogen in it is one or more, be further preferably methyl and/or halogen.
R2With R2' it is each independently selected from substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycle alkane Base, substituted or unsubstituted cycloalkenyl group, substituted or unsubstituted heterocycloalkenyl, substituted or unsubstituted condensed ring Aryl, substituted or unsubstituted condensed hetero ring base, substituted or unsubstituted aryl or substituted or unsubstituted miscellaneous Aryl;
R3With R3' be each independently selected from H, halogen, substituted or unsubstituted formamido, be substituted or do not taken The formic acid esters in generation, substituted or unsubstituted amino, substituted or unsubstituted nitro, cyano, acetonitrile-base, hydroxyl, hydrogen Sulfenyl, substituted or unsubstituted alkyl, substituted or unsubstituted miscellaneous alkyl;
R4With R4' be each independently selected from H, halogen, substituted or unsubstituted formamido, be substituted or do not taken The formic acid esters in generation, substituted or unsubstituted amino, substituted or unsubstituted nitro, cyano, acetonitrile-base, hydroxyl, hydrogen Sulfenyl, substituted or unsubstituted alkyl, substituted or unsubstituted miscellaneous alkyl;
R5With R5' it is each independently selected from H, halogen, substituted or unsubstituted amino, substituted or unsubstituted Nitro, cyano, acetonitrile-base, hydroxyl, sulfhydryl, substituted or unsubstituted alkyl, substituted or unsubstituted miscellaneous alkyl, Substituted or unsubstituted cycloalkyl, substituted or unsubstituted Heterocyclylalkyl, substituted or unsubstituted cycloalkenyl group, It is substituted or unsubstituted heterocycloalkenyl, substituted or unsubstituted fused ring aryl, substituted or unsubstituted thick miscellaneous Ring group, substituted or unsubstituted aryl, substituted or unsubstituted urea groups, substituted or unsubstituted phenoxy group, Or substituted or unsubstituted heteroaryl, substituted or unsubstituted ester group.
In the present invention, it is above-mentioned it is substituted in substituent group be selected from halogen, cycloalkyl, Heterocyclylalkyl, cycloalkenyl group, heterocycle Alkenyl, fused ring aryl, condensed hetero ring base, aryl, heteroaryl, formamido, formic acid esters, amino, nitro, cyano, acetonitrile-base, carboxylic It is one or more in base, sulfhydryl, alkyl, miscellaneous alkyl, phenoxy group, urea groups and ester group.
The virtual screening method and its compound of screening that the present invention is established have the following advantages that:1) by being established Virtual screening method reduces the quantity of experiment screening compound, shortens the R&D cycle, reduces financial cost;2) by vitro by Body radioassay is tested, and confirms virtual screening hit compound VSI-3, VSI-9, VSI-10, VSI-14, VSI-15, VSI- 16、VSI-17、VSI-18、VSI-20、VSII-2、VSII-7、VSII-17、VSIII-1、VSIII-8、VSIII-9、VSIII- 12、VSIII-15、VSIII-26、VSIII-28、VSIII-36、VSIII-37、VSIII-38、VSIII-43、VSIII-63、 VSIII-64 has preferable combination activity for diamondback moth ecdysone receptor.And VSIII-1, VSIII-8, VSIII-9, VSIII-12, VSIII-15, VSIII-26, VSIII-28, VSIII-37, VSIII-43 are under 500mg/L concentration, to diamondback moth With good killing activity, there is preferable application prospect in control of insect;3) moulting hormone obtained by virtual screening is similar Object structure novel is simple, is relatively easily-synthesized, low in raw material price, can further apply the exploitation of ecdysone agonists;4) virtual sieve Choosing hit compound VSI-1, VSI-2, VSI-3, VSI-4, VSI-5, VSI-6, VSI-7, VSI-8, VSI-10, VSI-12, VSI-15, VSI-16, VSI-19, VSI-20, VSI-23, VSIII-4, VSIII-39, VSIII-50 have mosquito larvae Good killing activity has preferable application prospect in health pest control.
In order to further illustrate the present invention, the screening to moulting hormone analog provided by the invention with reference to embodiments Method and lead compound are described in detail.
Reagent used in following embodiment is commercially available.
Embodiment 1
1) three-dimensional structure of 6 EcR/USP dimers and ligand eutectic is obtained from Protein Data Bank, respectively It is:Lepidoptera tobacco budworm EcR/USP dimers and ponasterone (PonA) (PDB:1R1K) and bishydrazide BYI06830(PDB:1R20);Homoptera Bemisia tabaci EcR/USP dimers and PonA (PDB:1Z5X);Elytrum hot eyes models one's literary or artistic style on that of the ancients robber EcR/USP dimers and PonA (PDB:2NXX);Lepidoptera tobacco budworm EcR/USP dimers and 20- hydroxyecdysones (20E) (PDB:2R40);Lepidoptera bollworm EcR/USP dimers and bishydrazide BYI08346 (PDB:3IXP).Using MOE Align/Superpose modules in 2014.08 compare six crystal complex ligand binding chamber positions of analysis, determine activity mouth Bag (Fig. 1).
2) using ligand in 6 crystal complexes of Ligand interaction module analysis in software MOE2014.08 Interaction between receptor extracts the common pharmacodynamic properties of natural ecdysone and bishydrazide compounds.4 naturally slough off All there are Thr343, Asn504, Ile339, Trp526 in the binding cavity of skin hormone, wherein Thr343, Asn504 does hydrogen bond confession Body, Ile339, Trp526 form hydrophobic region around the alkyl chain of natural ecdysone, and there are important hydrophobic effects.2 All there are Thr343, Tyr408, Ile339, Trp526, Met380 in the binding cavity of bishydrazide compounds, wherein Thr343 is done Hydrogen bond donor, Tyr408 do hydrogen bond receptor, and Ile339, Trp526, Met380 are respectively around the aromatic ring of bishydrazide, tertiary butyl Form two hydrophobic regions.
3) using Pharmacophore modules in software MOE2014.08, based on Ligand interaction modules Analysis result builds two Pharmacophore Models:Receptor-ligand Pharmacophore Model (Fig. 2), ligand Pharmacophore Model (Fig. 3).To test The separating capacity of Pharmacophore Model high low activity to compound is demonstrate,proved, by 25 high activity bishydrazide compounds, 14 low activities Bishydrazide compounds and 6 natural ecdysones are as test set.It is fixed at Thr343 in receptor-ligand Pharmacophore Model An adopted hydrogen bond donor site, Tyr408 places define a hydrogen bond receptor site, one hydrogen bond donor site of Asn504 definition, alkane A hydrophobic effect is defined at base chain, tertiary butyl.Positive accuracy=100% of receptor-ligand Pharmacophore Model, it is negative correct Rate=50%.In ligand Pharmacophore Model, Thr343, which corresponds at 17-C=O, 15-N on ligand, defines one as hydrogen bond Acceptor site, it is hydrogen bond donor site at 14-N that Tyr408, which corresponds on ligand, and Ile339, Trp526, Met380 correspond to and match The aromatic ring and alkyl of body are two hydrophobic effect sites.And excluded volume is added to binding pocket atom, optimize pharmacophore, exclude Volume radius R=1.72.Positive accuracy=100% of ligand Pharmacophore Model, negative accuracy=78%.
Screening marking is carried out to SPECS libraries using two Pharmacophore Models of structure, marking value S values are ranked up, S values It is more negative, show that the matching degree of micromolecular compound and pharmacophore is higher, 8851 compounds for selecting S values forward establish number According to library 1.
4) using the Calculate Descriptors modules in software MOE2014.08, database 1 is clustered and Quasi-medicated property is analyzed.Calculate weight (molecular weight), the ClogP (log of all compounds in database 1 octanol/water partition coefficient)、HBA(number of H-bond acceptor atoms)、HBD (number of H-bond donor atoms), ROB (number of rotatable bonds) and ARB (number of Aromatic atoms) parameter, according to class pesticide rule, select eligible:weight<435、ClogP<6、HBA<6、 HBD<2、ROB<9、ARB<17 micromolecular compound establishes database 2, totally 2538 compounds.
5) the docking agreement of acquiescence is selected to carry out the compound in database 2 using Dock modules in MOE 2014.08 Molecular docking.According to researchs of the Wang Ren little et al. to scoring functions, a variety of scoring functions combinations can improve molecular docking marking Reliability.The scoring functions high to select accumulation ability, we are by 6 compounds (tebufenozide, the furans worm acyl of commercialization Hydrazine, methoxyfenozide, chlorine tebufenozide, ring tebufenozide, suppression food hydrazine) and known 4 compounds effective to ecdysone receptor (BYI08346, BYI09181, BYI08738, BYI06934) is added to and picks out 1000 chemical combination from SPECS libraries at random Object forms accumulation ability evaluation library.Molecular docking is carried out by receptor of 1R20 to 1010 small molecules first.Molecular docking Afterwards, using the pose rescoring docking agreements in MOE to docking conformation S_A:GBVI/WSA dG、S_B:Affinity dG、S_C:Alpha HB、S_D:ASE、S_E:Five kinds of scoring functions of London dG are given a mark again.Then we are to difference Scoring functions and combination of function carry out accumulation ability evaluation (Fig. 4).It is found through experiments that, Affinity dG+Alpha HB+ The combination accumulation ability of tri- functions of ASE is most strong.Therefore molecular docking part in virtual screening, first we select acquiescence Docking agreement compound is docked, then with Affinity dG, Alpha HB, ASE scoring functions combination carry out Pose rescoring, and 32% compound establishes database 3 before choosing, totally 790 compounds.
6) compound activity prediction is carried out respectively using two methods:A) based on QuaSAR modules in MOE 2014.08; B) Conjugated free energy based on Amber12 calculates.
A) the QSAR models of bishydrazide compounds and its evaluation
According to structure diversity, 91 bishydrazide compounds composing training collection are selected, 41 compounds form test set. To test set compound using the AutoQSAR modules in MOE, iterative calculation is selected suitable descriptor combinations, is finally obtained Model eq (1).
Observations:91Descriptors:8 R2:0.64066 XR2:0.56736
Eq (1) model analysis:ASA-(negative accessible surface area)、E_tor(torsion energy)、PEOE_VSA+1(total positive 1 vdw surface area)、PEOE_VSA_NEG(total Negative vdw surface area), SlogP_VSA4 (4 slogp of bin (0.10,0.15)), a_hyd (number of hydrophobic atoms)、b_count(number of bonds)、vsurf_CW2(capacity factor at- 0.5)。
Model eq (1) is 36/41=87.8% to the accuracy rate that test set compound activity is predicted.So using eq (1) Activity Prediction is carried out to database 3, obtains 23 candidate compounds (VSI-1~VSI-23) of first virtual screening.
B) calculations incorporated free energy predicts receptor-binding activity
In the research of early period, MM-PBSA can preferably predict natural ecdyson and bishydrazide compounds with by The combination activity of body.First, by the compound of database 2, be rigidly docked in the binding cavity of 1R20, to 2538 compounds into Row 20ps Dynamics Optimizations, using the prediction of MM-PBSA methods and the combination of receptor activity.It is final to obtain second batch virtual screening 18 A candidate compound (VSII-1~VSII-18).
7) it to the ecdysone receptor combination determination of activity of 41 candidate compounds of two batches virtual screening, is turned over in-vitro transcription The pure protein (EcRB, USP) translated is receptor, and using the less salt buffer of PH=7.9 as buffer solution, setting competitive binding group is summarized It is combined, non-specific binding group, every group sets three repetitions.Competitive binding group:The diamondback moth ecdysone receptor working solution of 8 μ l (EcR+USP), 4.8 μ l [3H] PonA working solutions (final concentration 15nM), the 3.2 certain density untested compounds of μ l.Total binding Group (TB):The diamondback moth ecdysone receptor working solution (EcR+USP) of 8 μ l, 4.8 μ l [3H] PonA working solution (final concentrations 15nM), 3.2 μ l less salts buffer supplement volume to 16 μ l.Non-specific binding group:The diamondback moth ecdysone receptor work of 8 μ l Make liquid (EcR+USP), 4.8 μ l [3H] PonA working solutions (final concentration 15nM), the PonA, 16 μ of total volume of 3.2 μ l not tape labels l.Above-mentioned reaction solution 25 DEG C of reaction 90min in water-bath, reaction, which finishes, to be placed on ice.
Cylinder type filter (diameter 25mm, effective diameter 21mm) vacuum filtration separation, filter membrane select cellulose nitrate NC45, The ice-cold elution buffer filter washes film of 3ml 3 times, adds 3ml scintillation solutions, and liquid scintillation counter obtains data.Percentage bound is by as follows Formula is calculated, and experimental result is as shown in the figure (Fig. 5).The compound high for combining activity, gradient reduce concentration, measure IC50
8) structural similarity is retrieved:Guide's molecule VSI-9, VSI-10, VSII-14 based on hit carry out similitude inspection Rope obtains third batch virtual screening compound (VSIII-1~VSIII-64), and has carried out to diamondback moth ecdysone receptor knot Active testing is closed, obtains binding activity result figure such as figure of the compound to diamondback moth ecdysone receptor of third batch virtual screening Shown in 6.
According to the above results obtain having the compound of structure shown in Formulas I~formula IV or its pharmaceutically acceptable salt, ester, Solvate is respectively provided with diamondback moth ecdysone receptor and combines activity.
Wherein, R6、R6′、R11With R11' it is each independently H, alkyl, halogen;The alkyl is preferably the alkane of C1~C10 The alkyl of base, more preferably C1~C5;R7With R7' it is monosubstituted on phenyl ring, it is each independently alkyl, halogen;The alkyl The alkyl of the preferably alkyl of C1~C10, more preferably C1~C5;R8With R8' it is each independently H, acetonitrile-base, ester group, amide Base, halogen, alkyl;The alkyl is preferably the alkyl of C1~C10, more preferably the alkyl of C1~C5;The ester group and amide The carbon atom number of base is each independently 1~10, more preferably 2~10, is further preferably 2~5;R9With R9' H is each independently, Acetonitrile-base, ester group, amide groups, halogen, alkyl;The alkyl is preferably the alkyl of C1~C10, more preferably the alkane of C1~C5 Base;The carbon atom number of the ester group and amide groups is each independently 1~10, more preferably 2~10, is further preferably 2~5;R10 With R10' it is each independently 1,2 or 3 connected saturated or unsaturated five membered hydrocarbon ring or hexa-atomic hydrocarbon ring.
9) the general sieve of live body insecticidal activity is carried out to all obtained compounds of screening, obtains its insecticidal activity and the results are shown in Table 1, Part of compounds shows good mosquito-larvicidal activity as shown in Table 1;The desinsection for obtaining third batch virtual screening compound is lived Property it is general sieve the results are shown in Table 2, good killing activity is shown to diamondback moth under its 500mg/L concentration as shown in Table 2.
Compound is to the insecticidal activity of mosquito larvae under 1 various concentration of table
The general sieve of insecticidal activity of table 2 third batch virtual screening compound

Claims (10)

1. a kind of screening technique of moulting hormone analog, which is characterized in that including:
S1 the three-dimensional structure of EcR/USP dimers and ligand eutectic compound) is obtained from Protein Data Bank, is compared Analysis, determines active pocket;
S2 receptor-ligand Pharmacophore Model and ligand Pharmacophore Model) are built according to active pocket, and according to receptor-ligand medicine Effect group's model establishes database 1 with ligand Pharmacophore Model screening SPECS libraries, qualified compound;
S3) compound in the database 1 is clustered and quasi-medicated property is analyzed, qualified compound is selected to establish number According to library 2;
S4) using molecular docking programs, the compound in the database 2 with receptor is docked, establishes database 3, is used The receptor-binding activity of compound, obtains the compound of first virtual screening in QSAR model prediction garbled datas library 3;
S5) using the combination activity of compound and receptor in MM-PBSA methods prediction garbled data library 2, it is virtual to obtain second batch The compound of screening;
S6 the compound of first virtual screening and the compound of second batch virtual screening) are subjected to ecdysone receptor knot Close determination of activity, the lead compound hit;
S7 the lead compound) based on the hit carries out similarity retrieval to the compound in the SPECS libraries, obtains third Criticize the compound of virtual screening;
Wherein, the step S4) and step S5) have no point of sequencing.
2. screening technique according to claim 1, which is characterized in that the step S1) in compare analysis using in MOE Align/Superpose modules;The EcR/USP dimers are selected from Lepidoptera tobacco budworm EcR/ with ligand eutectic compound USP dimers and ponasterone eutectic compound, Lepidoptera tobacco budworm EcR/USP dimers and bishydrazide BYI06830 Eutectic compound, Homoptera Bemisia tabaci EcR/USP dimers and ponasterone eutectic compound;Elytrum hot eyes, which is modeled one's literary or artistic style on that of the ancients, steals EcR/ USP dimers are total to ponasterone eutectic compound, Lepidoptera tobacco budworm EcR/USP dimers and 20- hydroxyecdysones Brilliant compound, Lepidoptera bollworm EcR/USP dimers and bishydrazide BYI08346 eutectic compounds.
3. screening technique according to claim 1, which is characterized in that the step S2) in structure receptor-ligand pharmacophore Model and the method for ligand Pharmacophore Model are:EcR/USP dimers are extracted using Ligand interaction module analysis With the common pharmacodynamic properties of natural ecdysone in ligand eutectic compound and bishydrazide compounds, it is then based on common drug effect Feature is using Pharmacophore module construction receptor-ligand Pharmacophore Models and ligand Pharmacophore Model;The common drug effect Feature includes hydrogen bond receptor, hydrogen bond donor, excluded volume and hydrophobic effect.
4. screening technique according to claim 1, which is characterized in that the step S4) in molecular docking use Dock moulds Block carries out, and laying method is Triangle Matcher;After molecular compound is docked with receptor, using Affinity dG, The binding ability of Alpha HB, ASE scoring functions combination evaluation molecular compounds and receptor.
5. screening technique according to claim 1, which is characterized in that the QSAR models are established in accordance with the following methods:Choosing Take bishydrazide compounds composing training collection, the compound in database 3 forms test set, and the compound in test set is used AutoQSAR modules in MOE are iterated calculating, obtain QSAR models.
6. screening technique according to claim 1, which is characterized in that the step S5) it is middle using the prediction of MM-PBSA methods The Conjugated free energy of compound and the combination activity of receptor based on Amber12 calculates in garbled data library 2.
7. a kind of screening technique of moulting hormone analog, which is characterized in that including:
A1 the three-dimensional structure of EcR/USP dimers and ligand eutectic compound) is obtained from Protein Data Bank, is compared Analysis, determines active pocket;
A2) molecular compound in SPECS libraries is clustered and quasi-medicated property is analyzed, qualified compound is selected to establish number According to library 1 ';
A3 receptor-ligand Pharmacophore Model and ligand Pharmacophore Model) are built according to active pocket, and according to receptor-ligand medicine Effect group's model and ligand Pharmacophore Model garbled data library 1 ', qualified compound establishes database 2 ';
A4) using molecular docking programs, the compound in the database 2 ' with receptor is docked, establishes database 3 ', is used The receptor-binding activity of 3 ' middle compound of QSAR model prediction garbled datas library, obtains the compound of first virtual screening;
A5) using 2 ' middle compound of MM-PBSA methods prediction garbled data library and the combination activity of receptor, it is virtual to obtain second batch The compound of screening;
A6 the compound of first virtual screening and the compound of second batch virtual screening) are subjected to ecdysone receptor knot Close determination of activity, the lead compound hit;
A7 the lead compound) based on the hit carries out similarity retrieval to the compound in the SPECS libraries, obtains third Criticize the compound of virtual screening;;
Wherein, the step A1) and step A2) have no point of sequencing;The step A4) with step A5) have no successively it is suitable Point of sequence.
What 8. a kind of screening technique according to claim 1~7 any one obtained has killing activity for mosquito larvae Lead compound, have VSI-1, VSI-2, VSI-3, VSI-4, VSI-5, VSI-6, VSI-7, VSI-8, VSI-10, VSI- 12nd, structure shown in VSI-15, VSI-16, VSI-19, VSI-20, VSI-23, VSIII-4, VSIII-39, VSIII-50:
It is combined 9. what a kind of screening technique according to claim 1~7 any one obtained has with ecdysone receptor Activity lead compound, have VSI-3, VSI-9, VSI-10, VSI-14, VSI-15, VSI-16, VSI-17, VSI-18, VSI-20、VSII-2、VSII-7、VSII-17、VSIII-1、VSIII-8、VSIII-9、VSIII-12、VSIII-15、VSIII- 26th, structure shown in VSIII-28, VSIII-36, VSIII-37, VSIII-38, VSIII-43, VSIII-63, VSIII-64:
10. a kind of screening technique according to claim 1~7 any one obtain to diamondback moth have moulting hormone by Body combines the moulting hormone analog of activity and/or live body killing activity, which is characterized in that has formula one, formula two, formula three, formula The compound of structure shown in four or its pharmaceutically acceptable salt, ester, solvate:
Wherein, R1With R1' it is each independently selected from substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycle It is alkyl, substituted or unsubstituted cycloalkenyl group, substituted or unsubstituted heterocycloalkenyl, substituted or unsubstituted thick Cyclophane base, substituted or unsubstituted condensed hetero ring base, substituted or unsubstituted aryl or substituted or unsubstituted Heteroaryl;
R2With R2' be each independently selected from substituted or unsubstituted cycloalkyl, substituted or unsubstituted Heterocyclylalkyl, Substituted or unsubstituted cycloalkenyl group, substituted or unsubstituted heterocycloalkenyl, substituted or unsubstituted condensed ring virtue Base, substituted or unsubstituted condensed hetero ring base, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl Base;
R3With R3' it is each independently selected from H, halogen, substituted or unsubstituted formamido, substituted or unsubstituted Formic acid esters, substituted or unsubstituted amino, substituted or unsubstituted nitro, cyano, acetonitrile-base, hydroxyl, sulfhydryl, Substituted or unsubstituted alkyl, substituted or unsubstituted miscellaneous alkyl;
R4With R4' it is each independently selected from H, halogen, substituted or unsubstituted formamido, substituted or unsubstituted Formic acid esters, substituted or unsubstituted amino, substituted or unsubstituted nitro, cyano, acetonitrile-base, hydroxyl, sulfhydryl, Substituted or unsubstituted alkyl, substituted or unsubstituted miscellaneous alkyl;
R5With R5' be each independently selected from H, halogen, substituted or unsubstituted amino, substituted or unsubstituted nitro, Cyano, hydroxyl, sulfhydryl, substituted or unsubstituted alkyl, substituted or unsubstituted miscellaneous alkyl, is taken acetonitrile-base Generation or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl group, are taken substituted or unsubstituted Heterocyclylalkyl Generation or unsubstituted heterocycloalkenyl, substituted or unsubstituted fused ring aryl, substituted or unsubstituted condensed hetero ring base, Substituted or unsubstituted aryl, substituted or unsubstituted urea groups, substituted or unsubstituted phenoxy group are taken Generation or unsubstituted heteroaryl, substituted or unsubstituted ester group.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109712720A (en) * 2018-12-20 2019-05-03 周凌霄 The Mass production method of Pharmacophore Model
CN110246538A (en) * 2019-06-14 2019-09-17 北京市计算中心 Small molecule target nucleic acids aptamers computer-aided screening method and small molecule target nucleic acids aptamers based on high-performance calculation platform
CN116153412A (en) * 2022-12-14 2023-05-23 山西省人民医院 CHRNA7 ligand screening and application

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105820167A (en) * 2016-04-15 2016-08-03 南京工业大学 Aryl pyrazole purine compound with insecticidal effect, preparation method and application
CN106565678A (en) * 2016-11-10 2017-04-19 杭州市第人民医院 Virtual screening method for JAK2 kinase inhibitor and medicine lead compound

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105820167A (en) * 2016-04-15 2016-08-03 南京工业大学 Aryl pyrazole purine compound with insecticidal effect, preparation method and application
CN106565678A (en) * 2016-11-10 2017-04-19 杭州市第人民医院 Virtual screening method for JAK2 kinase inhibitor and medicine lead compound

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
HU XUEPING等: "Exploration of the binding affinities between ecdysone agonists and EcR/USP by docking and MM-PB/GBSA approaches", 《J MOL MODEL》 *
HU, XUEPING等: "Virtual screening for potential ligands of insect ecdysone receptors", 《ABSTRACTS OF PAPERS, 248TH ACS NATIONAL MEETING & EXPOSITION》 *
STN: "《CASRN:701930-92-9等》", 《STN REG》 *
WANG QINGMIN等: "Syntheses and Biological Activities of Novel N-tert-butyl-N′-Alkoxycarbonyl-N-Substituted Benzoylhydrazines and their Derivatives", 《LETTERS IN ORGANIC CHEMISTRY》 *
WANG QINGMIN等: "Synthesis and biological activity of novel N -tert-butyl-N -substituted benzoyl-N-(substituted phenyl)aminocarbonylhydrazines and their derivatives", 《TETRAHEDRON LETTERS》 *
傅晓东等: "芳基吡唑氟尿嘧啶类化合物的设计、合成与杀虫活性研究", 《有机化学》 *
胡雪萍等: "QSAR和MM-PBSA在EcR蜕皮激动剂虚拟筛选中的应用", 《第十届全国化学生物学学术会议论文摘要集(墙报)》 *
胡雪萍等: "蜕皮激素类似物的活性预测平台", 《中国化工学会农药专业委员会第十六届年会》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109712720A (en) * 2018-12-20 2019-05-03 周凌霄 The Mass production method of Pharmacophore Model
CN110246538A (en) * 2019-06-14 2019-09-17 北京市计算中心 Small molecule target nucleic acids aptamers computer-aided screening method and small molecule target nucleic acids aptamers based on high-performance calculation platform
CN110246538B (en) * 2019-06-14 2021-05-07 北京市计算中心 Small molecule target aptamer computer-assisted screening method based on high-performance computing platform and small molecule target aptamer
CN116153412A (en) * 2022-12-14 2023-05-23 山西省人民医院 CHRNA7 ligand screening and application
CN116153412B (en) * 2022-12-14 2023-07-28 山西省人民医院 CHRNA7 ligand screening and application

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