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Using Environment-Sensitive Fluorescent Probes to Characterize Liquid-Liquid Phase Separation in Supersaturated Solutions of Poorly Water Soluble Compounds

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Abstract

Purpose

Highly supersaturated aqueous solutions of poorly soluble compounds can undergo liquid-liquid phase separation (LLPS) when the concentration exceeds the “amorphous solubility”. This phenomenon has been widely observed during high throughput screening of new molecular entities as well as during the dissolution of amorphous solid dispersions. In this study, we have evaluated the use of environment-sensitive fluorescence probes to investigate the formation and properties of the non-crystalline drug-rich aggregates formed in aqueous solutions as a result of LLPS.

Methods

Six different environment-sensitive fluorophores were employed to study LLPS in highly supersaturated solutions of several model compounds, all dihydropyridine derivatives.

Results

Each fluoroprobe exhibited a large hypsochromic shift with decreasing environment polarity. Upon drug aggregate formation, the probes partitioned into the drug-rich phase and exhibited changes in emission wavelength and intensity consistent with sensing a lower polarity environment. The LLPS onset concentrations determined using the fluorescence measurements were in good agreement with light scattering measurements as well as theoretically estimated amorphous solubility values.

Conclusions

Environment-sensitive fluorescence probes are useful to help understand the phase behavior of highly supersaturated aqueous solutions, which in turn is important in the context of developing enabling formulations for poorly soluble compounds.

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Abbreviations

ASDs:

Amorphous solid dispersions

CMC:

Critical micelle concentration

DASPEI:

2-(4-(dimethylamino)styryl) -N-Ethylpyridinium Iodide

DMSO:

Dimethyl sulfoxide

ESIPT:

Excited state intermolecular proton transfer

HPMC:

Hydroxypropylmethyl cellulose

HPMCAS:

Hydroxypropylmethyl cellulose acetate succinate

ICT:

Intermolecular charge transfer

IPA:

Isopropyl alcohol

LLPS:

Liquid-liquid phase separation

PLM:

Polarized light microscopy

PRODAN:

4-(4-(Diethylamino)styryl) -N-Methylpyridinium Iodide (4-Di-2ASP) 6-Propionyl-2-dimethylaminonaphthalene

PVP:

Polyvinylypyrrolidone

TFA:

Trifluoroacetic acid

UV:

Ultraviolet

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ACKNOWLEDGMENTS AND DISCLOSURES

We would like to acknowledge AbbVie Inc. for providing research funding for this project.

Purdue University and AbbVie jointly participated in study design, research, data collection, analysis and interpretation of data, writing, reviewing, and approving the publication. Lynne S. Taylor is a professor at Purdue University, and has no additional conflicts of interest to report. David A. Alonzo is an employee at Gilead Sciences, Inc. and has no additional conflicts of interest to report. Shweta A. Raina, Geoff G. Z. Zhang and Yi Gao are employees of AbbVie and may own AbbVie stock.

Author information

Authors and Affiliations

  1. Department of Industrial and Physical Pharmacy, College of Pharmacy, Purdue University, 575 Stadium Mall Drive, West Lafayette, Indiana, 47907, USA

    Shweta A. Raina & Lynne S. Taylor

  2. Drug Product Development, Research and Development, AbbVie Inc, North Chicago, Illinois, USA

    David E. Alonzo, Geoff G. Z. Zhang & Yi Gao

  3. Manufacturing Science and Technology, AbbVie Inc, North Chicago, Illinois, USA

    Shweta A. Raina

  4. Formulation & Process Development, Gilead Sciences, Inc, Foster City, California, USA

    David E. Alonzo

  5. Analytical Sciences, Manufacturing Science and Technology, AbbVie Inc, North Chicago, Illinois, USA

    Yi Gao

Authors
  1. Shweta A. Raina
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Correspondence to Lynne S. Taylor.

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Raina, S.A., Alonzo, D.E., Zhang, G.G.Z. et al. Using Environment-Sensitive Fluorescent Probes to Characterize Liquid-Liquid Phase Separation in Supersaturated Solutions of Poorly Water Soluble Compounds. Pharm Res 32, 3660–3673 (2015). https://doi.org/10.1007/s11095-015-1725-z

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  • DOI: https://doi.org/10.1007/s11095-015-1725-z

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