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MOE's pharmacophore modeling tools determine the chemical features and their spatial
arrangement in 3D that are essential to the binding of a ligand to its receptor and thus to the
ligand's drug activity. Pharmacophore models can be generated from the structural data of
protein-ligand complexes as well as from ligands when no receptor information is available and
from the receptor structure when no ligands are available. The generated models can be used to
screen virtual compound libraries for potentially active molecules.
What is a Pharmacophore?
Creating Features
Consensus Features
Pharmacophore Elucidator
About Constraints
Running in MOE/batch
References
Pharmacophore Reference
SVL Commands
See Also
What is a Pharmacophore?
The term pharmacophore was coined by Paul Ehrlich in 1909 to mean "a molecular framework
that carries (phoros) the essential features responsible for a drug's (pharmacon) biological
activity" [Ehrlich 1909]. In other words, a pharmacophore is a 3D model describing the type and
location of the binding interactions between a ligand and its target receptor. The IUPAC
definition [IUPAC 1998] is more precise: "A pharmacophore is an ensemble of steric and
electronic features that is necessary to ensure the optimal supramolecular interactions with a
specific biological target and to trigger (or block) its biological response." The term
supramolecular means noncovalent. Typical pharmacophoric features include hydrogen bond
donor, hydrogen bond acceptor, hydrophobic, and positively and negatively ionized areas.
such as hydrogen donors and acceptors, aromatic centers, projected positions of possible
interaction partners or R-groups, charged groups, and bio-isosteres. The annotation points on a
ligand are the potential locations of the features that will constitute the pharmacophore query.
Annotation points relevant to the pharmacophore are converted into query features with the
addition of an extra parameter: a non-zero radius that encodes the permissible variation in the
pharmacophore query's geometry.
Once generated, a pharmacophore query can be used to screen virtual compound libraries for
novel ligands. Pharmacophore queries can also be used to filter conformer databases, e.g. output
from molecular docking runs, for biologically active conformations.
Pharmacophore Search. Search 3D conformation databases for structures that match the
pharmacophore query. The Conformation Import application can be used to generate 3D
conformation databases from vendor catalogs and in-house databases.
Model-Composer. Create a composite model from one or more models of varying types,
including pharmacophore.
Acetate
Acetic acid
Pharmacophore Annotations
Annotation is the process of identifying regions of pharmacophoric importance in space around a
molecular conformation and associating with those regions pharmacophore types. In MOE, the
Pharmacophore Query Editor both annotates the molecular system currently loaded in MOE as
well as renders the resulting annotations graphically in the MOE Window.
Locating and assigning pharmacophore annotations can be done in different ways. Although
MOE provides a variety of built-in pharmacophore annotation schemes or simply schemes, each
of which has its own particular set of annotation types, the default Unified scheme is to be
preferred as it is comprehensive, providing the annotation types of all the other schemes without
loss of efficiency.
Once assigned, annotations can be turned into features using the Pharmacophore Query Editor.
Note, however, that it is not a requirement that a feature be created from an annotation. Features
can be created, positioned, and assigned pharmacophoric types within the Pharmacophore Query
Editor independent of annotations, although it is rarely necessary to do so. Annotations thus
serve as suggestions for which type of features to use and where they should be located. In
almost all cases, features are created from annotation points.
H-bond
Donor
Acc
H-bond
Acceptor
Cat
Cation
Ani
Anion
ML
Metal
Ligator
Hyd
A
Hydrophobi
c Atom
Projected
Don2 Projected
Donor
Acc2
Projected
Acceptor
ML2
Projected
Metal
Ligator
PiN
Ring
Normal
Aromatic
PiR
pi-Ring
Hyd
Hydrophobi
c
CN2
NCN+
O2
COO-
Launch the Pharmacophore Consensus panel, which is used to suggest a consensus set of
pharmacophore features.
For more details, please refer to the Pharmacophore Query Editor Reference.
Pressing the Scheme: Info button in the panel opens a list of the different annotation types and
controls which ones are displayed in the Main Window. The Ligand Annotation options in the
bottom of the panel control which atoms are annotated (e.g. visible atoms or atoms of selected
chains only) and how the annotation points are displayed (e.g. as points, with or without labels).
For example, in Long mode, each annotation point is labeled by its annotation type.
Donor Atoms
Acc
Acceptor Atoms
Aro
Aromatic centroids
Hyd
Pharmacophore annotations
These may be isolated visually by turning off the display of all other types. This can be
accomplished using the Unified Info panel, which is opened with
Scheme: Info
This panel lists all the types of annotation points defined in the current scheme.
1. Turn off the display of all annotations.
Show | None
2. Turn on the display of the desired annotations.
Use the checkboxes in the Unified Info panel.
The HIV1-PR ligand should now be annotated with 4 kinds of annotation points.
Creating Features
A location in space, which can be modified directly in the MOE Window in the same way
as that of an atom: by selecting the feature (either in the panel or in the MOE Window)
and then using Shift-Alt-drag middle;
A logical expression called a feature expression that may include both annotation point
types and SMARTS strings. A feature inherits its types from its generating annotation
points.
Once completed, a pharmacophore query can be saved to a MOE pharmacophore file for future
use by pressing Save in the Pharmacophore Query Editor. The default extension is .ph4. This
file can subsequently be edited with the Pharmacophore Query Editor or used by other
applications such as Pharmacophore Search or Dock.
Example: Creating and Editing Pharmacophore Query Features for the 1HPV
Ligand
a. In the MOE Window, select the annotation point at the center of the ring. If two
annotation points overlap, select them both.
b. Press Create: Feature in the Pharmacophore Query Editor. A new feature will be
drawn in the MOE Window and will appear in the Pharmacophore Query Editor
list.
One of the rings will result in a feature labeled Hyd, and two in features labeled Hyd|
Aro.
2. Change the feature color.
Select all features in the list with Popup | Select All, then use the Color option to change
the color of all features to Green.
The features can also be edited singly by selecting them one at a time and making the
changes.
modifying the contents of the F* textfield to Hyd|Aro. Press Apply to put the change
into effect.
6. Save the pharmacophore query for future use.
Press Save. In the Write Query File panel, enter 1HPV-query1.ph4. Press OK.
The Pharmacophore Consensus panel is opened from the Pharmacophore Query Editor by
pressing Consensus (at top right). Once the suggested features have been calculated (by pressing
Calculate in the Pharmacophore Consensus panel), they can be transferred to the invoking
Pharmacophore Query Editor by selecting them in the Suggested Features list and pressing
Load Selected. The selected features will be appended to the feature list in the query editor.
Please refer to the Pharmacophore Tutorial for an example of how to use the
Pharmacophore Consensus application.
Pharmacophore Elucidator
The Pharmacophore Elucidator exhaustively generates pharmacophore queries for a set of
aligned molecules. and scores them based on a combined measure of how well they overlay the
ligands and how well they classify the data set into actives and inactives.
Molecular conformations can be either read in or calculated at runtime by choosing one of the
conformation generation methods from the Conformations option menu (As-Is implies that the
conformations have been precalculated).
To run the application, an Output Database name must be supplied. The Active Coverage
(which is the number of active molecules represented by the given query) and Feature Limit
(maximum number of pharmacophore features) parameters can be adjusted based on the amount
and type of data available. Query Spacing and Query Cluster are resolution tuning parameters
used to determine when a query is considered to be representative of a molecule and when two
queries are considered to be the same. Only different queries that satisfy the Active Coverage and
Feature Limit parameters are output.
For more details about scoring and query generation, please refer to the Pharmacophore
Elucidator manual page.
When invoked from a Pharmacophore Query Editor, the query currently open in the Editor is
used for the search. This launch point is convenient when the query is undergoing iterative
refinement, and search results are being examined after each modification step to evaluate the
changes being made. A Pharmacophore Search panel launched from a Pharmacophore Query
Editor automatically uses the latest version of the pharmacophore query for its search. The
databases to be searched are supplied when Search is pressed, and can be modified from within
the Pharmacophore Search panel using the Input controls.
If Pharmacophore Search is invoked from a Database Viewer, a pharmacophore query must be
specified in the Pharmacophore Search panel using the Query: Browse button. A
Pharmacophore Query Editor can also be opened from the Pharmacophore Search panel using
Query: Edit to edit the query currently being used by the Search panel.
In general, the bound conformations of the ligands being searched are not known. A
conformational search must be performed on the candidate molecules prior to initiating the
search (or existent conformational databases used); the pharmacophore search is then applied to
those conformations. The search proceeds by annotating each candidate ligand conformation and
then attempting to match the annotations to the constraints of the pharmacophore query such that
all restrictions of the query are satisfied. There can be more than one way in which a candidate
conformation can match a query.
Successful matches (hits) can be written to an output database using the Output options; they
can also be selected or isolated in the input Database Viewer using the Hit Entries menu.
search hits to be selected in the Database Viewer. Keep the default parameters in the
Results area of the panel: the output conformations will be written to the file
ph4out.mdb.
5. Perform the search.
Press Search to begin the pharmacophore search. The status line at the bottom of the
panel reports the current status of the search. Hits will be selected in the Database Viewer
as the search proceeds.
6. Examine the results.
When the search is finished, the status line reports the number of unique molecules
searched and the number of unique molecules of which at least one conformation was
found to be a hit. In this case, almost all the molecules are search hits. The hit structures
output to the database ph4out.mdb are aligned to the query. Press Results: Open to open
the output database in a Database Viewer.
The first structure from the database being viewed is automatically loaded into MOE.
The 1HPV-query1.ph4 pharmacophore query is rather simplistic and would probably match
many inactive compounds. At the same time, it successfully matched most of the candidate
inhibitors. This suggests that the query may be a reasonable starting point for a more advanced
query. In the next section, query refinement will be discussed.
only ligand information. If the receptor structure is available, knowledge of the binding pocket
can be used to enhance the query with steric and other structural constraints.
Some strategies for refining pharmacophore queries include:
Adding / deleting features and volumes. Larger numbers of features may reduce the hit
rate but increase specificity.
Determining features from pocket analysis. Examination of the binding pocket, for
example in terms of electrostatics and preferred contacts, can reveal potentially important
pharmacophoric features.
Adjusting feature radius and position. Custom fine-tuning can improve performance.
Note that features are normalized with respect to feature size (radius) so that large radius
features (i.e. features with higher positional tolerance) are not given more importance
than small radius ones.
When further refinement of a pharmacophore query using explicit 3D constraints yields little
gain, 2D modeling approaches in MOE can be used to refine the pharmacophore search results.
For example, a binary QSAR filter could be used to pre- or post-process the hit list.
Please refer to the Pharmacophore Tutorial for an example of how to refine a query using
interactive tuning and directional features.
About Constraints
MOE pharmacophore queries can be refined with a number of different constraints:
Volume and Shape Filters
Volume constraints are spatial constraints imposed on particular atoms. These constraints
serve both to confine hits to spatial regions in which steric clashes with the receptor are
avoided as well as to ensure that favorable contact regions will be populated with
appropriate atoms.
Each volume constraint, analogously to features, has an associated volume expression.
The volume expression is a SMARTS string. The empty string (i.e. if no expression is
specified) is equivalent to [#Q], which means "any heavy atom". The volume constraint
places restrictions only on atoms that match the volume expression.
There are three types of volume constraint:
The volume constraint is specified as one or more spheres where each sphere is centered
on a selected atom.
Tip Volume constraints are evaluated on atom centers, i.e the atoms are considered to be
points having zero radii. As a consequence, there still may be clashes between the hit
molecules and the pharmacophore volumes. It is advisable therefore to take into account
the van der Waals radii of candidate ligand atoms by calculating the Interaction (VDW)
surface and adjusting (increasing, if excluded volume, decreasing otherwise) the radii of
the volume constraints so that they just penetrate the interaction surface. To allow for
some tolerance, relax (decrease if excluded volume, increase otherwise) this radius by
0.3-0.5.
Feature Expressions
These are logical constraints that can be added to both features and volumes, e.g. Don &
Acc and can help focus the query toward a specific property.
SMARTS Matches
The chemistry of a group (substructure) to be matched can be specified within a feature
expression using SMARTS strings, e.g. Don & Acc ! "[N]"
Partial Matches
Allowing partial matches permits optional features to be included and can make the query
less specific.
Essential Features
Features marked essential to the query must always be matched, even when partial
matching is enabled.
Group Constraints
These specify that a certain number of the specified features must be matched. This kind
of constraint may be useful for accounting for alternative feature locations, e.g. to allow
for sidechain flexibility.
Please refer to the Pharmacophore Tutorial for an example of how to use volume
constraints.
In the Ligand Interactions panel, press Isolate to isolate the ligand and pocket residues in the
MOE Window. Turn on hydrogen bond display with
MOE | Footer | Contacts
A bound water HOH_201 (also bound to ILE50) and an H-bond interaction of the ligand
hydroxyl group with residue ASP25 can be observed in both the Ligand Interactions panel and
the MOE Window.
2.
Ligand and receptor, hydrogens hidden
In these images, the transparency of the surface has been adjusted (using the TF and TB
sliders in the Surfaces and Maps panel) for better ligand visibility.
4.
Calculate pocket electrostatics.
The electrostatic maps of the 1HPV pocket predict where hydrophobic, hydrogen bond
acceptor and hydrogen bond donor contacts are preferred. Partial charges must be
assigned to the system before the calculation is performed. Here, the charges have already
been assigned by the Protonate 3D application.
In the Surfaces and Maps panel, change the Surface to Electrostatic Map. Press
Apply.
The calculation solves the Poisson-Boltzmann equation for the electrostatic potential,
which is then used to generate the acceptor, donor, and hydrophobic predictive maps. The
maps show the locations in space at which an atom of the given type has a potential equal
to the value (in kcal/mol) given by the corresponding Level slider.
We will now examine each of the electrostatic maps in turn.
5.
Hydrophobic map
Analyze the hydrophobic map.
Isolate the hydrophobic map. The hydrophobic map gives the locations of receptor
contact and of low preference for acceptors and donors.
Here, the rendering mode has been set to Solid and the color to green. Hide the other two
maps by setting the rendering mode of Acc and Don to None.
There are four -3 kcal/mol hydrophobic regions. The 1HPV receptor comprises two
chains identical in sequence, and, accordingly, the hydrophobic areas appear as a
mirrored pair, with one smaller and one larger hydrophobic preference region associated
with each chain. These regions are suggestive of locations for hydrophobic and/or
aromatic pharmacophore features.
6.
Acceptor map
Analyze the hydrogen acceptor map.
Isolate the hydrogen acceptor map by setting the Hyd and Don rendering modes to None.
Here, the rendering mode of Acc has been set to Solid.
There is a -2 kcal/mol hydrogen acceptor region contacting the bound water HOH_201.
This suggests adding a projected feature to the pharmacophore to contact this area.
7. Analyze the hydrogen donor map.
Donor map
Isolate the hydrogen donor map by setting the Hyd and Acc rendering modes to None.
Here, the rendering mode of Don has been set to Solid.
There is a -2 kcal/mol hydrogen donor region at ASP25 contacting the hydroxyl group on
the ligand. This suggests a possible donor pharmacophore feature.
Refining the Query
Having analyzed the ligand-pocket interactions, we can now refine the 1HPV-query1
pharmacophore query by adding the pharmacophore features suggested by the analysis. If it is
still open, close the Ligand Interactions panel as it is no longer needed.
1.
Hydrophobic map and query
3.
Acceptor map with query
Add features suggested by the hydrophobic map.
Show the hydrophobic map (in isolation) by setting the Hyd rendering mode to Solid.
The hydrophobic features already in the query can be seen to coincide with the regions of
preferred hydrophobic contact. Thus, no additional features are needed to capture the
interactions depicted by the hydrophobic map.
4. Add features suggested by the hydrogen acceptor map.
Hide the hydrophobic map (set its rendering mode to None in the Surfaces and Maps
panel).
In the Pharmacophore Query Editor, press the Scheme: Info button to open the
information panel for the current scheme (Unified). In the Unified Info panel, turn on
Acc2 to enable the projected acceptor annotation points (the pharmacophore type is Acc2)
and then close the Info panel. Turn on Show Projecting Vectors in the Pharmacophore
Query Editor.
In the MOE Window, projected Acc2 acceptor annotation points will be displayed. It can
be seen that the projections from the C=O and SO2 groups coincide with the bound water.
Select these two annotation points, then press Create: Feature. An Acc2 projected
feature will be created at the centroid of the selected annotation points.
5.
Donor map with query
Add features suggested by the hydrogen donor map.
In the Unified Info panel, turn off Acc2. In the Pharmacophore Query Editor, change the
Ligand Annotation render mode to Long.
In the MOE Window, an annotation point can be seen on the hydroxyl group. Select this
annotation point (it is easiest to do this by left-dragging the mouse), then press Create:
Feature to create a Don&Acc feature on the hydroxyl group.
6.
Excluded volumes
Add excluded volumes on the receptor atoms of the binding site to avoid steric clashes
within the pocket.
First ensure no atoms are selected using MOE | Popup | Select | Clear. Then, select the
binding pocket atoms with MOE | Popup | Select | Pocket.
In the Pharmacophore Query Editor, press Create: Union. This will create a grouped
volume feature made up of volumes placed on all selected atoms. By default, the volume
is already of type Excluded.
Increase the radius R of the volume feature to 2.0. As explained above in the section on
constraints, atoms are considered to have zero radii when evaluating them against volume
constraints. To avoid clashes, the volume radius should be increased to take into account
the van der Waals radii of the ligand atoms of interest. The value of 2.0 accounts for the
atom radii, with some tolerance.
For easier visualization, you can turn off the display of the volumes in the MOE Window
by selecting V1 in the feature list of the Pharmacophore Query Editor and then turning on
the Hidden checkbox.
When the volume feature is selected in the list and the Hidden attribute is on, the centers
of the volumes are marked with selection markers and a dotted contour line drawn around
the volumes showing the volume's location.
Note that the Hidden attribute only affects rendering in the MOE Window. It has no effect
on searching: hidden features participate in pharmacophore searching as normal.
7. Save the refined query.
Press Save in the Pharmacophore Query Editor. In the Write Query File panel, enter the
name 1HPV-query2.ph4 and press OK.
The search results in 24 hits out of 82 molecules. Compared to the original query 1HPVquery1.ph4, the refined query is more selective. If the output database is examined as
described above in the example on pharmacophore search, it can be seen that the hits
exhibit mostly the same orientation, a desirable result.
Experiments using partial matches can be made. However, since the presence of the
projected acceptor feature Acc2 prevents hits that displace the bound water, the Acc2
feature should probably be made essential when partial matches are enabled. To make a
feature essential, turn on its F:Essential attribute in the Pharmacophore Query Editor.
Running in MOE/batch
Pharmacophore query generation in general requires user interaction and is not well-suited for
batch mode. Pharmacophore search, on the other hand, can be done in MOE/batch using an SVL
script or runnable file that invokes the SVL function ph4_Search. In the following, we show how
to use a script to run pharmacophore search as a batch job.
An SVL script is a sequence of SVL expressions stored in a plain text file. The expressions are
executed as though they had been entered at the SVL CLI (Command Line Interface), except that
in a script each expression must be terminated by a semi-colon (;).
Here is an example of a one-line script for performing a pharmacophore search on the database
dbfile.mdb using the pharmacophore query file query.ph4:
ph4_Search ['dbfile.mdb', 'query.ph4'];
The above call assumes that dbfile.mdb contains a pre-calculated molecule index field called
mseq (molecule sequence number).
Here is an example of a more complicated invocation, using some of the possible ph4_Search
options. These lines would be the exact contents of the script file:
ph4_Search [
'dbfile.mdb',
'query.ph4',
[
molfield: 'ligand2',
maxmolhits: 5000,
use_mseqfield: 0,
out_dbfile:'srch_out.mdb',
out_append: 0
//
//
//
//
];
Supposing one of the above calling sequences is saved to the file 'ph4search.svl'. The search
can then be executed in MOE/batch using the following in a command shell (Unix) or in a
command prompt window (Windows):
% moebatch -script ph4search.svl
This command invokes moebatch to run the script and then terminate once the script has finished
executing.
Using print and write statements in the script file allows additional information to be reported to
the shell:
ph4_Search ['dbfile.mdb', 'query.ph4'];
write ['\nDone. {}\n\n', asctime []];
References
[Ehrlich 1909] Ehrlich, P.; Ber. Dtsch. Chem. Ges. 42 (1909) 1747.
[IUPAC 1998] Wermuth, C.G., Ganellin, C.R., Lindberg, P., Mitscher, L.A.; Glossary of Terms
Used in Medicinal Chemistry (IUPAC Recommendations 1998); Pure & Appl.
Chem. 70:5 (1998) 11291143.
Pharmacophore Reference
Pharmacophore Tutorial. Use MOE pharmacophore applications to study serotonin 5HT3 receptors.
SVL Commands
ph4_Search
See Also
PLIF
Flexible Alignment
Site Finder
Surfaces and Maps
Dock
Pharmacophore-Type Molecular Fingerprints
Pharmacophore Feature Descriptors