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Afr. J. Biomed. Res. Vol.19 (September, 2016); 213- 218
Full Length Research Paper
Correlation of Lipophilicity Descriptors with Pharmacokinetic
Parameters of Selected Benzodiazepines
Adeyemo M.A1 and Idowu S.O1*
1
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Ibadan, Ibadan, Nigeria.

ABSTRACT
In early-stage drug discovery science, it is often important to reliably predict the absorption, distribution, metabolism and
elimination (ADME) property of new chemical entities in pipeline, in order to filter molecules that are not drug-like. Several in-
vitro models of lipophilicity profiling as predictor of ADME property have been developed. The validity of lipophilicity
determination based on different descriptors was evaluated using 4 model compounds of the benzodiazepine class; bromazepam,
clonazepam, diazepam and lorazepam. Lipophilicity descriptors describing the retention behaviours of the model compounds
were obtained from three approaches, namely; planar chromatographic determination (i.e. Isocratic chromatographic
hydrophobicity index), calculated log P values (i.e. clog P, ChemAxon), and octanol-water partition coefficient (i.e. log P). These
descriptors were correlated with in-vivo pharmacokinetic parameters - maximum plasma concentration (Cmax), time to reach peak
plasma concentration (Tmax) and area under plasma concentration – time curve (AUC), using the Pearson rank correlation. The
experimental approaches ranked diazepam as most lipophilic while in-silico approach ranked lorazepam as most lipophilic. AUC
and Cmax correlated positively with the lipophilicity descriptors while T max gave negative correlation (except for the in-silico
method) (r = 0.60, -0.74, and -0.25) with lipophilicity descriptors clog P, ICHI and logP respectively. ICHI gave the best
correlation with pharmacokinetic parameters - Cmax (r = 1.0), AUC (r = 0.89) and T max (r = -0.74) The planar chromatographic
platform was shown to be a valid biomembrane model for lipophilicity profiling. The lipophilicity descriptor, ICHI is positively
and strongly correlated with Cmax, and found superior to the correlation of Cmax with time-honoured octanol/water log P. A larger
study is thus warranted to delineate the potential of ICHI for general utility.

Keywords: Lipophilicity descriptors, benzodiazepines, pharmacokinetic parameters, correlation.

*Author for correspondence: E-mail: olakunleid@yahoo.com; Tel: +234-80-5842-7072

Received: April 2016; Accepted: July, 2016

Abstracted by:
Bioline International, African Journals online (AJOL), Index Copernicus, African Index Medicus (WHO), Excerpta medica (EMBASE), CAB
Abstracts, SCOPUS, Global Health Abstracts, Asian Science Index, Index Veterinarius

INTRODUCTION Lipophilicity is established as a physicochemical property

with major predictive value on ADMET properties and
The drug discovery process is a very rigorous, time consuming pharmacological activity, since molecules must traverse
and capital intensive venture in the pharmaceutical industry, biological membrane in order to interact with receptors and
because it is usually accompanied by a high degree of attrition. elicit drug action (Serda et al, 2012). It is reported to be
As a result several molecules fail to reach the market as drugs, closely correlated with permeability and drug solubility (Cross
with failure occurring even at the last phase of clinical trials. et al, 2003), hence it is widely used in pharmacokinetic
Attrition is often attributed to poor pharmacokinetic modeling (Kaliszan et al, 2003). Although lipophilicity
parameters -absorption, distribution, metabolism and toxicity enhancement of molecules improves bioavailability of drug, it
(ADMET) (Kennedy, 1997). Several physicochemical often leads to alteration of activity (Feng et al, 2010).
parameters such as lipophilicity, solubility, polar surface area, Lipophilicity refers to relative affinity of a molecule for
and permeability have been found to influence lipophilic environment, commonly measured by its
biopharmaceutical properties and drug likeness of a molecule partitioning in a biphasic medium (International Union of Pure
(Lipinski, 2000). These properties must be accurately and applied Chemistry, 1997). Various approaches used in the
predicted at the early stage of the drug discovery process, to determination include logarithm of octanol-water partition
reduce incidence of attrition (Kern et al, 2003; Penzotti et al, coefficient (log P), referred to as “the gold standard”
2004). (Mannhold et al, 2009); potentiometric titration techniques,
 Lipophilicity and Pharmacokinetic Parameters of Benzodiazepines

chromatographic techniques like reversed phase planar determined by inear regression analysis (GraphPad Prism
chromatography, high performance liquid chromatography, version 6.0, 2015). The x-intercept (φo) known as the ICHI,
immobilized artificial membrane chromatography (IAM), extrapolated from the graph was used in ranking the
immobilized liposome chromatography (ILC), micellar lipophilicity of the model compounds (Idowu et al, 2009).
electrokinetic chromatography (MEKC) and Bio-partitioning
Micellar Chromatography (BMC) etc. Correlation Analysis: Secondary data obtained from the
Lipophilicity is a critical parameter for central nervous literature (www.drugbank.ca) comprising of the experimental
system agents that are active in vivo (Ewelina, 2013). The octanol - water partition coefficient and calculated log P using
benzodiazepines are among the most commonly prescribed the most popular and widely accepted ChemAxon clog P
centrally acting drugs used as anxiolytics, muscle relaxants, algorithm were pooled together with the experimentally
antiepileptic and as hypnotic agents (Mihic and Harris, 2011); determined chromatographic lipophilicity descriptor, ICHI.
whose pharmacology depends on the compound’s This was correlated with pharmacokinetic parameters like
lipophilicity. Lipophilicity of benzodiazepines has been area under the plasma concentration-time curve (AUC), peak
reported to influence their speed of action; and thus the plasma concentration (Cmax) and time to reach peak plasma
rational choice of the more lipophilic members of the class e.g. concentration (Tmax) which were culled from literature. The
diazepam in eliciting rapid effect in acute conditions (Vinkers correlation analysis of the three lipophilicity descriptors with
et al, 2012). Furthermore, the absorption pharmacokinetic the pharmacokinetic parameters was performed using the
profile indicated that the highly lipophilic benzodiazepines Pearson correlation coefficient as a measure of goodness-of-
penetrate the CNS rapidly with consequent tendency for high correlation between the parameters. All statistical analysis
abuse (Melton and Kirkwood, 2014). was performed by GraphPad Prism version 6.0 for Windows
The validity of lipophilicity measurement methods is (Graph Pad Software, San Diego, CA, www.graphpad.com,
dependent on the extent of similarity of the system to the 2015)
biological membrane architecture (i.e. biomimetic feature)
(Smith et al., 1996). In this paper, we report a comparative RESULTS
evaluation of the correlation of various lipophilicity
descriptors with pharmacokinetic parameters, as a validation The retention behavior of the model compounds depicted by
of the strategies adopted in obtaining the descriptors, using a the linear regression of Rm versus the organic modifier fraction
small chemical library of benzodiazepines (Figure 1). is displayed in Figure 2. The derived lipophilicity parameter
(Isocratic chromatographic hydrophobicity index –ICHI), and
MATERIALS AND METHODS experimental log P values and cLogP values (obtained from
the literature) for the model compounds are shown in Table 1.
Chemicals and Equipment: Diazepam, clonazepam, Correlation analysis between the pharmacokinetic parameters
lorazepam and bromazepam (Secondary reference sample, (Table 2) revealed different levels of correlation with the
British Pharmacopoeia), pre-coated silica gel thin layer lipophilicity descriptors. Figure 3 shows the correlation of the
chromatographic plate (0.2mm, Merck, Germany) , n-hexane lipophilicity descriptors with the bioavailability parameter
(Analar, British Drug House), methanol (Analar, British Drug (AUC) revealing a poor correlation with experimental log P (r
House), liquid paraffin (Analar, British Drug House), Mettler =0.47), negative correlation with clog P (r = -0.47) and fair
H80 analytical balance (UK), UV lamp (254 and 365 nm, correlation with chromatographic ICHI parameter (r = 0.89).
Gallenkamp, U.K.) Correlation between the maximum plasma concentration
Preparation of reversed phase stationary phase: 5% liquid (Cmax) and time for maximum plasma concentration (Tmax) is
paraffin in n-hexane was used to coat the surface of the silica displayed in Figure 4 and 5 respectively.
gel plates (5 x 10cm) by the ascending development method.
Table 1: Lipophilicity index from planar chromatographic
Preparation of the mobile phase: Various concentrations (φ) experiment (ICHI), experimental log P (octanol/water partition) and
comprising of 30, 35, 40, 45, 50, 55, 60, 62.5 and 65% of calculated logP (ACD ChemAxon)
methanol-water mixture was prepared in developing the Lipophilicity descriptors
chromatogram of the model compounds. Model ICHI LogP cLogP
Lipophilicity profiling of the model compounds: Methanolic compound Experimental
solutions (2 µL of 1% w/v) of the model compounds were Bromazepam 0.529 2.05 2.54
spotted on reversed phase plates, and developed in the gradient Clonazepam 0.531 2.41 3.15
Diazepam 0.618 2.82 3.08
series of mobile phase. The retardation factor (Rf) obtained
Lorazepam 0.516 2.39 3.53
was transposed into Rm values (Biagi et al, 1969). Linear plots
of the Rm versus φ were made and the line of best – fit was

Table 2: Mean values of pharmacokinetic parameters of Bromazepam, Clonazepam, Diazepam and Lorazepam (Culled from literature)
AUC (nghr/ml) Cmax (ng/ml) Tmax (hrs)
Bromazepam 2501 (Podilsky et al, 2009) 46 (Podilsky et al, 2009) 1.53 (Podilsky et al, 2009)
Clonazepam 561(Crevoisier et al, 2003) 50 (Sio et al, 1975) 2.5 (Crevoisier et al, 2003)
Diazepam 4430 (Guo et al, 2013) 178 (Guo et al, 2013) 1.25 (Guo et al, 2013)
Lorazepam 405 (Blin et al, 1999) 20 (www.accessdata.fda.gov) 2.37 (Blin et al, 1999)

214 Afr. J. Biomed. Res. Vol. 19, No.3 (September) 2016 Adeyemo and Idowu
 Lipophilicity and Pharmacokinetic Parameters of Benzodiazepines

Figure 1:
Chemical structures of the study compounds

DISCUSSION

The lipophilicity parameter for the model compounds

obtained from the retention behavior on the chromatographic
platform displayed in Figure 2 and Table 1 revealed that
lipophilicity of the model compounds follows the sequence
below:
Diazepam > clonazepam > bromazepam > lorazepam
The ranking from the experimentally determined log P in a
octanol-water partition system followed a similar order but
slight disparity in the ranking of the last 2 compounds as
shown below:
Diazepam > clonazepam > lorazepam > bromazepam.
The ranking on the computer algorithm shows:
Lorazepam > clonazepam > diazepam > bromazepam.

The lipophilicity of benzodiazepines homologues has been

reported to vary with polarity and electronegativity of the
various substituents (Brunton et al, 2008). Lipophilicity
encodes different intermolecular forces, expressed as interplay
of the hydrophobic and polar interactions (Liu et al, 2011).
According to Van de Waterbeemd, the relationship between
Lipophilicity, hydrophobicity and polarity is:
Lipophilicity = Hydrophobicity – polarity

Figure 2:
Linear regression of the Rm versus the organic modifier Figure 3:
fraction revealing the retention behavior of the model Correlation analysis of the AUC with the 3 lipophilic
compounds descriptors

215 Afr. J. Biomed. Res. Vol. 19, No.3 (September) 2016 Adeyemo and Idowu
 Lipophilicity and Pharmacokinetic Parameters of Benzodiazepines

Figure 5:
Figure 4: Correlation analysis of Tmax with the 3 lipophilicity descriptors.
Correlation analysis of Cmax with the 3 lipophilicity descriptors
Nitrogen being a hydrogen bond acceptor, will confer water
This implies that the greater the polarity of a molecule, the solubility, thus making bromazepam less lipophilic than
lower the lipophilicity. Thus, based on structural theory, clonazepam, devoid of an heterocycle albeit with a polar nitro
lorazepam which has a hydroxyl moiety (a hydrogen bond substituent on another phenyl ring. The ranking based on the
donor and acceptor) in the benzodiazepine skeleton is results of the chromatographic method thus corroborate
expected to have the lowest lipophilicity, while diazepam structural theory. On the contrary, the in-silico approach (c log
which has an additional methyl group should be the most P) emphasized the inherent limitation of in-silico approach in
lipophilic. predicting molecular properties, due to inability of computer
Between the two extremes, bromazepam has a nitrogen algorithms to fully account for the 3 dimensional structural
heteroatom in a 6-membered heterocycle (pyridine moiety) details of molecules and lack of parameterization of certain
instead of the phenyl ring in all the other compounds. molecular fragments (Mannhold, 2008). Ranking lorazepam

216 Afr. J. Biomed. Res. Vol. 19, No.3 (September) 2016 Adeyemo and Idowu
 Lipophilicity and Pharmacokinetic Parameters of Benzodiazepines

as most lipophilic of the series cannot be supported by drug discovery science. Such a larger study is ongoing in our
structural theory. The closeness of values used in ranking the laboratory.
experimentally determined parameters implies they are more
accurate estimate of lipophilicity than the in silico predictions. Acknowledgement
(Eros et al, 2002) The authors thank Ms. Monsurat Oriyomi Nureni for her
technical assistance
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