Supercritical fluid chromatography approach for a sustainable manufacture of new stereoisomeric anticancer agent

Article abstract of DOI:10.1016/j.jpba.2017.08.006

Two routes aimed at the manufacture of unprecedented stereoisomeric combretastatin A-4 analogue were described: flash chromatography vs supercritical fluid chromatography. The latter has many advantages over liquid chromatography and was therefore chosen for the small scale separation of methyl 1-[(3-hydroxy-4-methoxyphenyl) (3,4,5-trimethoxyphenyl)methyl]-5-oxo-L-prolinate 5, with potential antitumoral activity. After a screening of six different polysaccharide based chiral stationary phases and four co-solvents, the percentage of co-solvent, the flow-rate and the outlet pressure were optimized through a design of experiments (DoE). The preparation of 50 mg of each stereoisomer was achieved successfully on a Chiralpak AD-H with isopropanol as a co-solvent. Productivity (kkd), solvent usage and environmental factor (E Factor) were calculated. Flash chromatography and supercritical fluid chromatography approaches were compared in terms of yield and purity of each stereoisomer manufactured.

Full text of DOI:10.1016/j.jpba.2017.08.006

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Accepted Manuscript
Title: Supercritical fluid chromatography approach for a
sustainable manufacture of new stereoisomeric anticancer
agent
Authors: Alina Ghinet, Yasmine Zehani, Emmanuelle Lipka
PII:
S0731-7085(17)31417-6
DOI:
Reference:
http://dx.doi.org/doi:10.1016/j.jpba.2017.08.006
PBA 11450
To appear in:
Journal of Pharmaceutical and Biomedical Analysis
Received date:
Revised date:
Accepted date:
1-6-2017
2-8-2017
3-8-2017
Please cite this article as: Alina Ghinet, Yasmine Zehani, Emmanuelle Lipka,
Supercritical fluid chromatography approach for a sustainable manufacture of
new stereoisomeric anticancer agent, Journal of Pharmaceutical and Biomedical
Analysishttp://dx.doi.org/10.1016/j.jpba.2017.08.006
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Supercritical fluid chromatography approach for a sustainable manufacture of new
stereoisomeric anticancer agent
Alina GHINET1, 2, 3, Yasmine ZEHANI1, 4, Emmanuelle LIPKA 1,4
*
1 Univ. Lille, Inserm, U995 - LIRIC - Lille Inflammation Research International Center, F-59000 Lille, France
2
Ecole des Hautes Etudes d’Ingénieur (HEI), Laboratoire de Pharmacochimie, 13 rue de Toul, F-59046 Lille,
France
3 ‘Alexandru Ioan Cuza’ University of Iasi, Faculty of Chemistry, Bd. Carol I nr. 11, 700506 Iasi, Romania
4 UFR Pharmacie, Laboratoire de Chimie Analytique, BP 83, F-59006 Lille, France
GRAPHICAL ABSTRACT
Highlights
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Stereoisomeric combretastatin A-4 analogue as anticancer agent
Two different approaches were applied for diastereomeric separation.
Optimization of percentage of co-solvent, flow-rate and outlet pressure in pSFC.
Green factors evaluation
Abstract
Two routes aimed at the manufacture of unprecedented stereoisomeric combretastatin A-4
analogue were described: flash chromatography vs supercritical fluid chromatography. The
latter has many advantages over liquid chromatography and was therefore chosen for the small
scale separation of methyl 1-[(3-hydroxy-4-methoxyphenyl) (3,4,5-trimethoxyphenyl)methyl]-
5-oxo-L-prolinate 5, with potential antitumoral activity. After a screening of six different
polysaccharide based chiral stationary phases and four co-solvents, the percentage of co-

Evaluation Only. Created with Aspose.PDF. Copyright 2002-2021 Aspose Pty Ltd.
solvent, the flow-rate and the outlet pressure were optimized through a design of experiments
(DoE). The preparation of 50 mg of each stereoisomer was achieved successfully on a Chiralpak
AD-H with isopropanol as a co-solvent. Productivity (kkd), solvent usage and environmental
factor (E Factor) were calculated. Flash chromatography and supercritical fluid
chromatography approaches were compared in terms of yield and purity of each stereoisomer
manufactured.
Keywords: Diastereoisomers; chirality; productivity; kkd; E factor
Introduction
The development of drugs marketed as single enantiomers is growing continuously in the
pharmaceutical business and in academic laboratories alike, as it can be observed from the
number of single enantiomers launched in recent years [1]. At the discovery stage, the most
important factor is time. The required amount usually ranges between a few milligrams to 50 g
and the cost factor as well as the scale-up feasibility are negligible. However, it is important to
be able to isolate the single enantiomers in a short period of time in order to rapidly perform
the biological tests. Over the past fifty years high performance liquid chromatography (HPLC)
has become very popular, particularly for preparative scale. However, faster analysis time
demands of the market, together with the necessity of a greener and sustainable way of
separation issued by governmental authorities have facilitated a supercritical fluid
chromatography (SFC) renew of these last 15 years. Taking advantage of its intrinsic properties
[2] this technique became the mainstream one in preparative scale. In SFC, the mobile phase is
constituted by, at least, 50% of supercritical carbon dioxide (from T = 31°C and P = 74 bar).
The low viscosity and high diffusivity of the mobile phase together with a lower pressure drop
permit high flow-rate with a reduced influence on peak efficiency. Carbon dioxide is not polar
2

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enough [3] and most of the time a polar modifier such as alcohol or acetonitrile must be added
to elute chiral pharmaceutical compounds. It is worth noting that the addition of a liquid
modifier moves conditions to subcritical, however advantages stated previously still exist.
Among the widely known twelve principles of green chemistry stated by Anastas and Warner
in 1998 [4] not all, but many, can be directly applied to analytical chemistry: prevent waste, use
safer solvents and processes, minimize use of energy, avoid chemical derivatives, analyze in
real time to prevent pollution and lastly increase the safety of the operator [5]. Some papers
have addressed this topic of green chromatography in practice [6-9]. Nevertheless, three of the
principles, called the three Rs: Reduce, Replace, Recycle are considered to be most relevant for
greening both the analytical [10-11] and large scale preparative separation technologies [12]
and can be applied daily in every laboratory [6]. Based on those trends, SFC has established it-
self as a green method, in particular for the small-scale preparative, mainly because of the
numerous advantages of CO2.
Indeed the replacement of hazardous organic solvent by a large amount of carbon dioxide is
desirable in an eco-friendly point of view. It is non-toxic, non-flammable, and a renewable
resource as the SFC installation uses carbon dioxide that is condensed from the atmosphere (or
industrial waste plumes) delivered into the chromatographic device and then returned to the
atmosphere (or recycled). It is important to indicate that preparative SFC is not a net generator
of carbon dioxide. This gas is a recovered industrial waste byproduct (such as the production
of cement, the production of metals or alloys such as iron and steel, and the production of
chemicals) or comes from natural processes like beverage fermentation, while incineration of
organic solvents results in the net generation of carbon dioxide [13].
From an economical point of view, replacing the mobile phase by large carbon dioxide amount
reduces expensive organic solvent volume to be used, and evaporated (energy cost). With
regard to HPLC, which consumes large volume of water (in Reversed Phase) needing high
3