CN104744375A - Guanidine NTR1 micromolecular antagonist - Google Patents

Guanidine NTR1 micromolecular antagonist Download PDF

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Publication number
CN104744375A
CN104744375A CN201510069216.8A CN201510069216A CN104744375A CN 104744375 A CN104744375 A CN 104744375A CN 201510069216 A CN201510069216 A CN 201510069216A CN 104744375 A CN104744375 A CN 104744375A
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ntr1
ring
small molecular
compound
guanidine class
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CN201510069216.8A
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CN104744375B (en
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徐峰
张果
李晓丹
王坤
田园诗
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Nankai University
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Nankai University
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D235/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings
    • C07D235/02—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings condensed with carbocyclic rings or ring systems
    • C07D235/04—Benzimidazoles; Hydrogenated benzimidazoles
    • C07D235/24—Benzimidazoles; Hydrogenated benzimidazoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached in position 2
    • C07D235/30—Nitrogen atoms not forming part of a nitro radical

Abstract

The invention provides a guanidine NTR1 micromolecular antagonist and relates to the fields of drug design science and pharmacology. Compounds having a general formula I as shown in the specification are provided by virtue of virtual screening, cell activity testing and structural modification based on a receptor, wherein R1 represents a hydrophobic group; R2 represents a substituted aromatic ring; R3 represents a polar substituted group. The compounds have inhibition activity on NTR1 and have important use in preparing drugs for treating NTR1 protein related diseases.

Description

A kind of guanidine class NTR1 small molecular antagonists
Technical field
The present invention relates to medicinal design and field of pharmacology, test in particular to through virtual screening and cytoactive, and carry out the structure of a series of guanidine class NTR1 micromolecular inhibitors obtained based on the structural modification of acceptor.
Background technology
G protein coupled receptor (G Protein-Coupled Receptor, GPCR) has seven transmembrane structure, extensively distributes in vivo, participates in the response activity to various signaling molecule.GPCR and signal transduction system thereof are in running balance usually, and start the adjustment of downstream signaling pathway according to the change of various physiologic factor, maintain the stable of intracellular environment; Under pathological conditions, cell accepts outer signals by GPCR equally and regulates cell activities.Especially in malignant cell, GPCR is by the signal transduction process of mediate tumor cell, the process such as generation, growth, invasion inhibition of tumour cell is regulated, almost in all malignant cells, the improper expression of certain class GPCR molecule can both be detected.The excessive activation of GPCR or suppression all may cause the imbalance of intracellular signal transduction approach, thus cause disorderly physiological response, cause abnormal physiological phenomenon.
In GPCR superfamily, category-A member neurotensin receptor 1 (NTR1) albumen is mainly distributed in central nervous system and gastrointestinal system, Gq protein signaling path in mediated cell, the concentration level of calcium ion in regulating cell.The native agonist neurotensin (NT) of this albumen is 13 peptides, and wherein six amino acid Arg-Arg-Pro-Tyr-Ile-Leu play a crucial role.Discovered in recent years NT and NTR1 plays crucial effect in the growth of Several Kinds of Malignancy (mammary cancer, carcinoma of the pancreas, colorectal carcinoma, lung cancer, prostate cancer etc.), propagation, invasion and attack, transfer.There is bibliographical information in the cancerous tissue of normal mammary epithelial and infiltrative breast carcinoma, all have the expression of NT, and NTR1 only high expression level in the cancerous tissue and tumor cell line of mammary cancer, in infiltration ductal carcinomas of breast, NTR1 expression rate is up to 91%.Animal experiment study confirms that NTR1 can promote the growth of transplanted tumor cell and can shorten the oncocyte doubling time, the Infiltration and metastasis of inducing transplantation knurl.Therefore, the antagonist of NTR1 can play certain restraining effect to the generation development of tumour, is expected to the active drug becoming the diseases such as treatment tumour.The crystalline structure of mouse source NTR1 was successfully parsed in 2012, and this is provide favourable condition based on the medicinal design of acceptor and the Optimizing Reconstruction of compound.
This laboratory to have set up early stage one for NTR1 containing 10000 micromolecular databases, effectively improve breakneck acceleration.At present, in document, report multiple antagonist for NTR1 target, from known antagonist, contributed to the screening method setting up Pharmacophore Model and 2D-Fingerprint, thus find new small molecule antagonist.
Summary of the invention
Existing is that the new drug development of target spot not yet occurs marketed drug with NTR1, and is not resolved due to the crystalline structure of NTR1 at that time, and the three-dimensional structure that can only obtain according to homology modeling carries out drug screening and design.The present invention seeks to the three-dimensional crystalline structure based on the mouse source NTR1 albumen of having resolved, carry out virtual screening to incorporating the NTR1 small molecules database that multiple compound database obtains and cytoactive test is carried out to the selection result, structural modification based on acceptor is carried out to the activated small molecules of tool, to seek micromolecular compound NTR1 to inhibit activities, for the preparation of the medicine of the disease relevant to NTR1.
The present invention tests and structural modification through virtual screening and cytoactive, and obtain a kind of guanidine compound, structure is as shown in formula I.
Wherein:
R 1morpholine ring, do not replace and methyl, halogen, hydroxyl replace C 5-C 6cycloalkyl group, piperidine ring, phenyl ring, pyridine ring, dimethylamino, sec.-propyl, tertiary butyl hydrophobic group; R 2the phenyl ring or pyridine lopps aromatic nucleus that are replaced by halogen, methyl or ethyl; R 3hydrogen, or amino, hydroxyl, aminomethyl or methylol polar group; N is 1-3.
During guanidine compound provided by the invention can be applicable to prepare with the medicine of NTR1 protein related diseases.
The present invention screens and is R through the micromolecular compound 1 with inhibit activities that cytoactive test obtains 1morpholine ring, R 2contraposition chlorine substituted benzene ring, R 3be hydrogen, n is 2, its structure following (compound 1), IC 50value is 44.49 μMs.
Advantage of the present invention and beneficial effect:
The present invention is by virtual screening and cytoactive test, and result proves, screening of the present invention obtains NTR1 small molecular antagonists (compound 1) has better inhibit activities to intracellular calcium concentration.And then analyzed for acceptor, charge property etc. by the hydrophilic and hydrophobic of the pocket to NTR1 and ligand binding, hydrogen bond, the result of docking with NTR1 according to compound 1 and marking, additions and deletions and the replacement of group is carried out at some position, and carry out docking checking, thus structure of modification and modification are carried out to compound 1, obtain a based compound, these compounds are by the NTR1 that acts in body thus participate in its regulation mechanism, for the preparation of the medicine of the diseases such as treatment or adjuvant therapy of tumors, Parkinson disease, schizophrenia, there is good medicinal application prospect.
Accompanying drawing explanation
Fig. 1 is the interaction schematic diagram of compound 1 and NTR1.
Fig. 2 is the interaction schematic diagram of compound 2 and NTR1 after modifying
Fig. 3 is the Ca of compound 1 2+fluorescent value is to the graphic representation of compound concentration logarithm.
Embodiment
Be further described with feature to various aspects of the present invention below.But these embodiments do not mean that has any restriction to the present invention.
The virtual screening of embodiment 1, NTR1 micromolecular inhibitor and cytoactive test
One, the method for virtual screening of the present invention comprises following 4 steps:
The structure of step 1:NTR1 small molecules database
Integrate multiple business small molecules database and (comprise ZINC, SPECS, J & K Screening Library, InterBioScreening, Timtec, Msdiscove, Iris-biotech, TCM Database, Ambinter NaturalProducts, AnalytiCon Discovery NP, Princeton NP, Molecular Diversity PreservationInternational, PUBCHEM), the molecule of repetition and the molecule of structural similitude is therefrom rejected with Pipeline pilot software, and pick out good absorption according to the ADMET character of the molecule calculated, the compound that toxicity is low carries out Cluster, obtain 1,000,000 micromolecular databases.By the Glide module in Schrodinger, these small molecules are carried out docking of SP precision with NTR1, choose the small molecules of marking front 10000 in result, form NTR1 small molecules database.
Step 2: the structure of Pharmacophore Model and screening
Find out from BindingDB database 6 activity better, compound (as follows) that textural difference is larger is as training set, the Create Pharmacophore Automatically module in Discovery Studio3.5 is utilized to set up Pharmacophore Model, collection checking and FitValue value determine most suitable Pharmacophore Model after tested, and this model comprises 2 hydrophobic centers, 1 aromatic nucleus center, 1 hydrogen bond donor.Search 3D Database is adopted to screen NTR1 small molecules database.
Step 3:Fingerprint screens
Obtain in BindingDB 3 active compounds (structural formula is as follows) and 11 non-active compounds are adopted respectively the Fingerprint module in Discovery Studio3.5,2D-Fingerprint is carried out with NTR1 small molecules database, the compound of structural similitude in structure in active compound Fingerprint result and non-active compound Fingerprint result is removed, more therefrom selects structure without the compound delivered and report.
Step 4: cluster analysis
The Cluster module of being screened by Fingerprint in pharmacophore model discrimination in step 2 and step 3 under the Design and Analyze Libraries in the result DiscoveryStudio3.5 obtained carries out cluster analysis, be divided into into 99 classes, from each class, select 1 molecule, obtain 99 compounds.Buy these compounds and cytoactive test is carried out to it.
Two, cytoactive testing method of the present invention comprises following 2 steps:
Step 1:NTR1 surely turns the structure of expressing cho cell system
With the RPMI1640 substratum containing 10% foetal calf serum, 37 DEG C, 5%CO 2, under the condition of saturated humidity, cultivate Chinese hamster ovary celI.By the DH5 α glycerol stock containing PUC57-NTR1 recombinant plasmid (restriction enzyme site is EcoR I and Hind III), transform, amplification, extract recombinant plasmid PUC57-NTR1, double digestion PUC57-NTR1 and expression vector pcDNA3.1 (+), reclaim goal gene NTR1 and expression vector pcDNA3.1 (+), NTR1 with pcDNA3.1 (+) is connected, connect product conversion DH5 α competence bacterium, positive colony is filtered out with penbritin, amplification, extracts plasmid, called after pcDNA3.1 (+)-NTR1.With pcDNA3.1 (+)-NTR1 Transfected Recombinant Plasmid Chinese hamster ovary celI, use a Chinese hamster ovary celI for transfection pcDNA3.1 (+) expression vector to make positive control simultaneously.The Chinese hamster ovary celI screening stable transfection with G418 is cloned, and go down to posterity enlarged culturing, obtains the people source system of mono-clonal.
Step 2: compound surely turns the active testing of Chinese hamster ovary celI to NTR1
NT is the native ligand of NTR1, after NT and NTR1 combines, the rising of intracellular calcium concentration can be caused by the signaling pathways of the Gq protein activation of coupling, the micromolecular compound having an inhibit activities to NTR1 then can the rising of calcium ion concn in T suppression cell, therefore after can adding NT and micromolecular compound by detection, whether intracellular calcium ion concn raises, and judges whether micromolecular compound has inhibit activities to NTR1.SR48692 is the selective antagonist of the NTR1 reported, can the activity of antagonism NT.
Adopt calcium 6 test kit, first carry out preliminary experiment.Add the NT of 1nM in 96 orifice plates after, add 10 μMs and 100 μMs of testing compounds respectively, measure the calcium current value of each compound under 10 μMs and 100 μMs of concentration, be chosen at the compound that 10 μMs have inhibit activities, its general IC of initial guess 50value.Then with this IC 50centered by value, draw 8 concentration gradients in the ratio geometric ratio of 1:2, the calcium current value of compound under mensuration different concns, and respectively using NT and SR48692 as negative and positive control, obtain calcium ion fluorescent value-compound concentration curve.The IC of GraphPad Prismversion5.0 computed in software micromolecular compound is adopted according to this curve 50value.
With above-mentioned virtual screening and cell measuring method for activity, screen and obtain the micromolecular compound (compound 1) NTR1 to inhibit activities: 1-(4-chlorophenoxy)-3-{2-imines-3-[2-(4-morpholine) ethyl]-2,3-dihydro-1-benzimidazolyl-}-2-propyl alcohol, its IC 50value is 44.49 μMs (Fig. 3).
Embodiment 2, carry out structural modification based on the mode of action of small molecules and acceptor NTR1.
First in Discovery Studio3.5, open the three-dimensional crystalline structure of NTR1 albumen, this structure is from PDB (ProteinData Bank), and No. PDB is 4GRV.Hydrogenation is carried out to it, anhydrates, leave out the T4 Lysozyme structure of insertion, adopt Prepare Protein module to prepare albumen, and find avtive spot with Define Site according to the NT in part.Prepare Ligands module is adopted to prepare small molecules.Ready, adopt the Libdock method of Dock Ligands module to dock, docking result as shown in Figure 1.According to this docking result, compound 1 forms hydrogen bond with key amino acid Arg327, Tyr146 of the acceptor of bibliographical information, and the bond distance of hydrogen bond is respectively (Fig. 1), this hydrogen bond action for the two in conjunction with extremely important.Chlorine substituted benzene chlorine in ring is replaced with sec.-propyl (compound 2), the hydrogen-bond length of this two places key shortens to respectively (Fig. 2).Bond distance is shorter, and hydrogen bond action is stronger, and the combination of part and acceptor is tightr, then the activity of part may be better.Adopt the cytoactive testing method in embodiment 1 to carry out survey to the compound 2 after modification to live, and calculate its IC 50value.Record the IC of compound 2 50value is 39.62 μMs.
Here be modified to example with this, be concrete implementation method is described, but can not as restriction.

Claims (7)

1. a guanidine class NTR1 small molecular antagonists, has such as formula the structure shown in I,
Wherein, R 1it is hydrophobic group; R 2it is substituted aroma ring; R 3hydrogen or polar substitution group.
2. the guanidine class NTR1 small molecular antagonists according to claims 1, is characterized in that, described hydrophobic group is morpholine ring, do not replace and C that methyl, halogen, hydroxyl replace 5-C 6cycloalkyl group, piperidine ring, phenyl ring, pyridine ring, dimethylamino, sec.-propyl, the tertiary butyl.
3. the guanidine class NTR1 small molecular antagonists according to claims 1, is characterized in that, described substituted aroma ring is the phenyl ring or pyridine ring that are replaced by halogen, methyl or ethyl.
4. the guanidine class NTR1 small molecular antagonists according to claims 1, is characterized in that, described polar substitution group is amino, hydroxyl, aminomethyl or methylol.
5. the guanidine class NTR1 small molecular antagonists according to any one of claims 1 to 4, is characterized in that, the n in described general formula is 1-3.
6. the guanidine class NTR1 small molecular antagonists according to claims 1, it is characterized in that, described compound is preferably R 1morpholine ring, R 2contraposition chlorine substituted benzene ring, R 3be hydrogen, n is 2, and its structure is as follows,
7. guanidine class NTR1 small molecular antagonists according to claim 1 is in preparation and the application in the medicine of NTR1 protein related diseases.
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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003105779A2 (en) * 2002-06-17 2003-12-24 Taktix Corporation Guanidine compounds as anesthetics and for treatment of nervous system disorders
WO2008003752A1 (en) * 2006-07-07 2008-01-10 Neurosearch A/S 2 -amino benzimidazole derivatives and their use as modulators of small - conductance calcium -act ivated potassium channels
CN101405000A (en) * 2006-01-19 2009-04-08 艾博特公司 2-imino-benzimidazoles
KR20090084502A (en) * 2008-02-01 2009-08-05 한국화학연구원 Antibacterial composition containing 2-iminobenzoimidazole derivatives
EP1838306B1 (en) * 2005-01-11 2010-11-17 NeuroSearch A/S Novel 2-amino benzimidazole derivatives and their use as modulators of small-conductance calcium-activated potassium channels
CN102307862A (en) * 2008-12-08 2012-01-04 贝林格尔.英格海姆国际有限公司 Compounds for treating cancer
WO2013033037A2 (en) * 2011-08-26 2013-03-07 The Regents Of The University Of California Novel antiprion compounds

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003105779A2 (en) * 2002-06-17 2003-12-24 Taktix Corporation Guanidine compounds as anesthetics and for treatment of nervous system disorders
EP1838306B1 (en) * 2005-01-11 2010-11-17 NeuroSearch A/S Novel 2-amino benzimidazole derivatives and their use as modulators of small-conductance calcium-activated potassium channels
CN101405000A (en) * 2006-01-19 2009-04-08 艾博特公司 2-imino-benzimidazoles
WO2008003752A1 (en) * 2006-07-07 2008-01-10 Neurosearch A/S 2 -amino benzimidazole derivatives and their use as modulators of small - conductance calcium -act ivated potassium channels
KR20090084502A (en) * 2008-02-01 2009-08-05 한국화학연구원 Antibacterial composition containing 2-iminobenzoimidazole derivatives
CN102307862A (en) * 2008-12-08 2012-01-04 贝林格尔.英格海姆国际有限公司 Compounds for treating cancer
WO2013033037A2 (en) * 2011-08-26 2013-03-07 The Regents Of The University Of California Novel antiprion compounds

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
RENE RAPHEMOT ET AL: "Discovery, characterization, and structure-activity relationships of an inhibitor of inward rectifier potassium (Kir) channels with preference for Kir2.3, Kir3.X, and Kir7.1", 《FRONTIERS IN PHARMACOLOGY OF ION CHANNELS AND CHANNELOPATHIES》 *
RENE RAPHEMOT ET AL: "Eliciting Renal Failure in Mosquitoes with a Small-Molecule Inhibitor of Inward-Rectifying Potassium Channels", 《PLOS ONE》 *
张果，等: "人源神经降压素受体-1稳转细胞系的建立及其应用", 《药学学报》 *

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