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Simultaneous diastereo- and enantioseparation of farnesoid X receptor (FXR) agonists with a quinine carbamate-based chiral stationary phase

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Abstract

In the frame of a project aimed at finding non-steroidal farnesoid X receptor (FXR) agonists, we identified 4-(2,4-dimethoxyphenyl)-3,6-dimethyl-1-(2-tolyl)-4,8-dihydro-1H-pyrazole[3,4-e][1,4]thiazepin-7-one (1) as a hit endowed with FXR activity. Most of the compounds synthesised during the hit-to-lead optimisation work were characterised by the presence of two chiral centres and were therefore obtained as mixtures of anti(±)- and syn(±)-diastereoisomers. A restricted sub-set of species harboured with a carboxylic acid group on the distal phenyl ring of the biphenyl (a(±)5 (A1) and s(±)5 (S1)) or the phenoxyphenyl (a(±)6 (A2) and s(±)6 (S2)) moiety at C-4 position of the pyrazole[3,4-e][1,4]thiazepin-7-one core, resulted in suitable diastereo- and enantioresolution with a quinine (QN) carbamate-derived chiral stationary phase (CSP). Differently from the compounds usually analysed with QN-based CSPs, the couples A1/S1 and A2/S2 were atypical selectands, in which the two chiral carbon atoms reside at a remote position with respect to the carboxylic function, the main “point of attack” to the CSP. We produced evidence that the scarcely employed normal-phase (NP) eluent systems represent the elective choice for achieving the simultaneous diastereo- and enantioseparation of this class of compounds over the usually preferred reversed-phase (RP) and polar-organic (PO) modes of elution. Indeed, after the optimisation of the eluent composition, NP conditions allowed to obtain profitable enantioselectivity profiles, along with excellent diastereoselectivity levels (α(A1) = 1.07, R S(A1) = 1.15; α(S1) = 1.09, R S(S1) = 1.47; α(A2) = 1.08, R S(A2) = 1.31; and α(S2) = 1.06, R S(S2) = 1.18). The optimised NP methods are suitable for simultaneously providing information on the diastereo- and enantiopurity of the investigated compounds.

Simultaneous diastereoand enantioseparation of two non-steroidal FXR agonists with a quinine carbamate-based chiral stationary phase, in the normal-phase mode of elution.

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Abbreviations

AA:

Ammonium acetate

ACN:

Acetonitrile

AcOH:

Acetic acid

BA:

Bile acid

BzA:

Benzoic acid

CDCA:

Chenodeoxycholic acid

CEA:

Cyclohexylamine

CECA:

Cyclohexanecarboxylic acid

DEA:

Diethylamine

DEAA:

Diethylammonium acetate

DIPEA:

N,N-diisopropylethylamine

ETN:

Ethanolamine

EtOH:

Ethanol

FA:

Formic acid

FXR:

Farnesoid X receptor

IPA:

2-propanol

MeOH:

Methanol

NP:

Normal phase

PO:

Polar organic

QN:

Quinine

RP:

Reversed phase

TEA:

Triethylamine

TEAA:

Triethylammonium acetate

TFA:

Trifluoroacetic acid

THF:

Tetrahydrofuran

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Acknowledgements

This work was supported by Intercept Pharmaceuticals (New York). We thank Professor Wolfgang Lindner (University of Vienna) for the generous gift of the column.

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Authors and Affiliations

  1. Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Via del Liceo 1, 06123, Perugia, Italy

    Roccaldo Sardella, Maura Marinozzi, Federica Ianni, Antonella Lisanti & Benedetto Natalini

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  1. Roccaldo Sardella
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  2. Maura Marinozzi
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  3. Federica Ianni
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  4. Antonella Lisanti
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  5. Benedetto Natalini
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Correspondence to Benedetto Natalini.

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Published in the special issue Analytical Science in Italy with guest editor Aldo Roda.

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Sardella, R., Marinozzi, M., Ianni, F. et al. Simultaneous diastereo- and enantioseparation of farnesoid X receptor (FXR) agonists with a quinine carbamate-based chiral stationary phase. Anal Bioanal Chem 405, 847–862 (2013). https://doi.org/10.1007/s00216-012-6348-1

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