PHASE: a new engine for pharmacophore perception, 3D QSAR model development, and 3D database screening: 1. Methodology and preliminary results

We introduce PHASE, a highly flexible system for common pharmacophore identification and assessment, 3D QSAR model development, and 3D database creation and searching. The primary workflows and tasks supported by PHASE are described, and details of the underlying scientific methodologies are provided. Using results from previously published investigations, PHASE is compared directly to other ligand-based software for its ability to identify target pharmacophores, rationalize structure-activity data, and predict activities of external compounds.

Authors and Affiliations

1. Schrödinger, Inc., 120 W. 45th St., 29th Floor, New York, NY, 10036, USA

   Steven L. Dixon, Alexander M. Smondyrev, Eric H. Knoll, Shashidhar N. Rao, David E. Shaw & Richard A. Friesner

2. Department of Chemistry, Columbia University, New York, NY, 10027, USA

   Eric H. Knoll & Richard A. Friesner

3. D E Shaw & Co, 120 W. 45th St., 39th Floor, New York, NY, 10036, USA

   David E. Shaw

Corresponding author

Correspondence to Steven L. Dixon.

Appendix A: selectivity estimation

In PHASE, the selectivity of a pharmacophore hypothesis H is defined as follows:

where p(H) is the probability that a random drug-like molecule will match the hypothesis, irrespective of any activity exhibited by that molecule toward the biological target in question. Given a database of drug-like molecules, it is straightforward to search that database for matches to a hypothesis, and thereby arrive at an estimate of selectivity based on that particular sample population of molecules. However, application of such a procedure is far too time-consuming to be practical when scoring a large number of hypotheses, so a rapid means of estimating selectivity based on the physical characteristics of a hypothesis is sought.

Van Drie [12] has shown that selectivities of two-point pharmacophores can be reliably estimated with respect to a given database using pre-tabulated probabilities that cover discrete distance ranges. He went on to show that highly selective three-point pharmacophores can be constructed by combining two-point pharmacophores with the highest selectivities. This is a natural consequence of the fact that the probability of matching a k-point pharmacophore \(\hbox{H}^{\langle k \rangle}\) is less than or equal to the probability of matching all (k· (k−1))/2 two-point pharmacophores embedded within \(\hbox{H}^{\langle k \rangle}\):

Strict equality is not preserved because a given molecule may match each of the two-point pharmacophores even if it fails to contain a single arrangement of k features that matches \(\hbox{H}^{\langle k \rangle}\). Nevertheless, since matching the two-point pharmacophores is a necessary condition for matching \(\hbox{H}^{\langle k \rangle}\), the right-hand-side of Eq. A2 is of interest for purposes of estimating selectivity.

If the two-point probabilities are independent, then the following relation holds:

Further, if sites i and j are separated by a distance of d _ij , and their pharmacophore feature types are α(i) and α(j), respectively, then Eq. A3 can be rewritten in terms of probabilities of matching specific inter-feature distances to within a tolerance Δd:

Given a population of drug like molecules and a pair of feature types x and y, there is a probability density p ^*(d _xy ) that describes the distribution of xy pharmacophores within that population. While p ^*(d _xy ) may be complex and possibly discontinuous, for purposes of estimating selectivity a simple Gaussian dependence is assumed, so that the probability density may be written as:

For small values of Δd, the following approximation can be made:

Substituting A5 and A6 into A4 yields

Taking logarithms,

Although it is certainly possible to estimate the univariate parameters σ_{α (i)α (j)} and μ_{α (i)α (j)} for each possible pair of feature types, it is advantageous to treat the right-hand-side of Eq. A8 as a general polynomial in d _ij , and fit the associated coefficients to observed probabilities for a large number and variety of pharmacophores:

This treatment can help overcome certain deficiencies in the model, such as the assumption that the two-point probabilities are independent of each other (Eq. A3). In practice, the second-order terms in Eq. A9 do not add much statistically independent information to the model, and we have found a first-order approximation to be satisfactory:

To determine appropriate values for the A and B parameters, a training set was assembled by randomly selecting 1000 minimized structures from a conformational database of the World Drug Index [36], then randomly choosing between two and seven pharmacophore sites from each structure. This yielded a training set of 1000 pharmacophores containing varying numbers of sites and different combinations of the features A, D, H, N, P, and R. A sample probability was computed for each pharmacophore H_λ by determining the number of structures M _λ out of the original 1000 that matched the pharmacophore to within a tolerance of 2.0 Å in all intersite distances:

Since there were six types of features in the sampled pharmacophores, the number of unique feature pairs was 21, requiring a total of 42 adjustable parameters. No attempt was made to optimize all of these independently because of the possibility of only limited information for certain pairs of features. For example, pharmacophores that contain both negative and positive ionizable features tend to be very rare among drug-like structures, so they cannot be expected to be well-represented in a relatively small population sample. Therefore, parameter values were determined by applying a partial least-squares (PLS) procedure to fit the −log₁₀(H_λ) values in terms of latent factors constructed from the pool of 42 variables. Details of the PLS algorithm used in PHASE are provided in Appendix B.

To arrive at an appropriate number of PLS factors to include in the model, predictions were made for a test set of 500 pharmacophores drawn from the same sample population of 1000 WDI structures. As successively more PLS factors were incorporated into the model, test set errors trended downward until reaching a minimum at 23 factors. At this point, the test set RMSE was 0.372 log units and Q ² was 0.786. This compared to a training set RMSE of 0.343 and R ² of 0.826. This model has been integrated into PHASE for computation of the Selectivity_Score term that appears in Eq. 7.

It is worth noting that training sets containing as many as 5000 structures were also investigated, and no significant improvement in the test set predictions was observed. The protocol of using 1000 structures was adopted because it is far less computationally demanding, and therefore represents a practical approach for users who wish to calibrate selectivity models based on a different set of structures.

Appendix B: partial least-squares regression

PHASE utilizes a standard recursive procedure for extracting orthogonal latent factors from a data matrix in a predetermined number of steps. It is distinguished from the NIPALS algorithm [37, 38], which is an iterative approach with a user-defined stopping criterion, but no absolute control over the total number of steps.

Let X∈R ^{n × m} represent the independent variable data matrix for a training set of n observations and a pool of m variables. Let y∈R ^{n × 1} represent the training set dependent data, which will be estimated using latent factors extracted from X. Creation of the PLS regression model proceeds as follows:

Center each column of X:

Center y:

Determine PLS factors and regression coefficients for up to M PLS factors (M ≤ m):

For a regression with M PLS factors, the estimates \({\hat{{\bf y}}}\) are then given by:

To apply the M-factor PLS model to a new set of \({\tilde{n}}\) observations with data matrix \({\tilde{{\bf X}}}\) , the regression coefficients b must first be translated back to the space of the original X variables:

Define

The coefficients b ^x may then be used to make estimates for the new observations as follows:

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