-Cyclodextrins as a Solubilization Tool

for
Insoluble APIs

One of the main challenges in drug delivery is to guarantee the bioavailability of the
active ingredients, especially when they are poorly soluble and/or with a limited
gastrointestinal permeability. To improve
aqueous solubility and thus the
therapeutic effectiveness of such compounds, a number of formulation strategies
have already been largely tested. It is a big challenge to find the right solubilization
tool for these active ingredients.
We will first explain why to choose -Cyclodextrins as a solubilization tool, and all
you need to know to successfully complexate your active ingredient with
-Cyclodextrins. Then using concrete examples will ease the understanding of the
tools used to study inclusion on -CDs: phase solubility, stability constant, and
complexation efficiency, complexation processes in liquid and solid state.
The Biopharmaceutics Classification System (BCS) predicts oral drug absorption
based on the chemicals aqueous solubility and intrinsic permeability through the
gastrointestinal mucosa. Two studies have been realized with highly permeable
drugs with limited water solubility, considered as BCS class II : Zotepine and
Zaleplon. These studies demonstrate why the Roquettes KLEPTOSE range
(chemically modified Beta Cyclodextrin) is the best solution for improving solubility
enhancement.

(-CDs) -Cyclodextrins are cyclic oligosaccharides with a bucket-like structure having a

hydrophobic internal cavity and a hydrophilic exterior. This unique structure allows for the
formation of inclusion complexes, where lipophilic compounds are non-covalently bound within
the cavity.

-Cyclodextrins (-CDs) are used to:

Increase

aqueous solubility of APIs

Increase chemical and physical stability of APIs
Reduce toxicity (e.g: local irritation after topical or oral
administration)
Enhance drug delivery to and through biological
membranes
Mask bad taste or odor
Convert liquid drugs to amorphous powders

-Cyclodextrins available on the market:

Native -cyclodextrin
-cyclodextrin (B-CD, KLEPTOSE, Roquette)
Modified -cyclodextrin
Hydroxypropyl--cyclodextrin with a 0.9 degree of molar
substitution (HP-CD, KLEPTOSE HP, Roquette)
Hydroxypropyl-- cyclodextrin with a 0.62 degree of molar
substitution (HPB-CD, KLEPTOSE HPB, Roquette)
-Cyclodextrin Sulfobutyl Ethers Sodium Salts (SBE)
Methyl--cyclodextrin (KLEPTOSE Crysmeb, Roquette
product under development showing interesting inclusion
properties)

What do you need to know about your API to successfully complexate it with
cyclodextrins (CDs)?
Small molecules:
how many atoms contains:
how many condensed rings:
water solubility:
melting point temperature:
molecular weight:
electrostatic charge:
log P
pKa
stability issues:

more than 5 atoms (C, P, S, and N) to form the skeleton of

the drug molecule
less than 5
less than 10mg/ml
below 250 C
between 100 and 400
positive, negative, neutral

chemical, photo, etc.

Large molecules:
the side chain in macromolecules may contain suitable groups which can interact with CDs in
aqueous solutions and form a partial complex with CDs.
Source: Lovatt,M., Cooper,A. and Camilleri,P. (1996) Eur. J. Biophys., 24, 354357

How are the complexes formed?

In the presence of water, CDs (host molecule) can form inclusion complexes with many drugs
(guest molecule) by taking up the molecule, or more frequently some lipophilic part of the
molecule, into the central cavity by a steric and a thermodynamic interaction. Drug molecules
in the complex are in rapid equilibrium with free molecules in the solution. No covalent bonds
are formed or broken during the complex formation.
Source: Frmming and Szejtli: Cyclodextrins in Pharmacy Kluwer Acad. Press, Dordrecht, 1994.

What type of inclusion complexes can -CDs form with an API?

What forces are involved in the complex formation?

Release of enthalpy-rich water molecules from the cavity

Electrostatic interactions
Van der Waals interactions
Hydrophobic interactions
Hydrogen bonds
Release of conformational strain and charge-transfer interactions

What solubilization solution can -CDs offer?

API formulation in Liquid phase and as Solid Dispersions

CASE STUDIES:
Using concrete examples will ease the understanding of the tools used to study inclusion on
-CDs: phase solubility, stability constant, and complexation efficiency, complexation processes
in liquid and solid state:

Case study 1: Zotepine as Model Drug for -CDs Solubilization

Case study 2: Zaleplon as Model Drug for Cyclodextrin Solubilization

Case Study 1: Zotepine as Model Drug for -CDs Solubilization

Zotepine is an atypical antipsychotic drug used in the treatment of
acute and chronic schizophrenia via blocking dopamine D2 receptors in
the central nervous system
Commercially available as a tablet in 25mg, 50mg and 100mg
Brand name : Nipolept, Losizopilon, Lodopin, Setous, Zoleptil
Due to its low solubility and extensive first pass metabolism its oral
bioavailability is 7-13%
Physical-chemical properties
Mol. Wt. : 331.86
Log P: 5.6
BCS class II (Low Solubility and High Permeability)
Zotepine is equipotent and price competitive compared to other
antipsychotics and its solubility enhancement by native and modified
betacyclodextrins can facilitate a better marketability.

What is the phase solubility?

Phase solubility evaluation (liquid state solubilization) using Zotepine as a model API
The phase solubility assessment is designed based on Higuchi and Connors method (T.
Higuchi and K.A. Connors: Phase-solubility techniques. Adv.Anal.Chem.Instrum.4, 117-212,
1965) (Fig. 1). Excess amount of API is added to aqueous solutions of increasing CDs
concentrations (10mM up to 50mM (or 200mM) and deionized water (pH 6.5) and optional
depending on the formulation requirements (API pKa) in buffer as controls (Fig.1 and Fig.3).
The vials are mixed for 1, 3 and 7 days at RT (25C) in order to evaluate the mixing time
needed to reach solubilization saturation. Aliquots of these solutions are filtered through
0.45m and analyzed by HPLC. Then the concentration of the solubilized API (expressed in
mM or M) is plotted for each CD concentration (in mM or M) and the resulting graph is
representing APIs Phase Solubility curve in the respective CD (Fig.3). Changes in
temperature and pH can lead to a different profile of the Phase Solubility Curve.

Fig.1 Vials set up for Phase Solubility evaluation at RT (25C) in deionized water (pH 6.5).

Fig.2. Samples set up for HPLC solubility evaluation, stability evaluation and lyophilization

Fig 3. Phase-solubility profiles of Zotepine in BCD,HPBCD, Crysmeb and SBE at RT (25oC) in deionized water (pH 6.5).

What profile the phase solubility curve can be?

Does phase solubility profile prove the inclusion complex formation?

The phase-solubility profiles do not verify formation of inclusion complexes. They only
describe how the increasing cyclodextrin concentration influences drug solubility.
Because cyclodextrins are also known to form:
- Non-inclusion complexes
- Complex aggregates
able to dissolve drugs through micelle-like structures.

What is Stability Constants (K1:1) and Complexation Efficiency (CE)?

From the Phase Solubility curve Fig 3. we can calculate as per Eq1 and Eq 2:

Where m is the slope of Phase Solubility curve as determined by linear regression and S0 is the
drug solubility in DI water or buffer as determined after (1 to 7) days of mixing (depending at what
time point the saturation solubility is reached).

Table 1. Zotepine Solubility Enhancement at RT (25oC) in deionized water (pH 6.5).

Table 2. Zotepine complexes K 1:1 and CE at RT (25oC) in deionized water (pH 6.5).

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Do we need stability evaluation of the API solubilized by -CDs in liquid

phase?
API solubilization is useless if the complex is not stable.
As per Fig 2 samples corresponding to each CD molarity in Phase Solubility experiment are put
on stability at 25 C and 40 C for 30 days and 60 days in order to evaluate API:CD complexes
stability in aqueous phase (Fig. 4).

Fig. 4. Zotepine complex stability evaluation at 25C and 40C.

If API:CD complexes are not stable in liquid phase do we have an alternative?

Yes, we lyophilize the liquid phase and we reconstitute it in a liquid vehicle just before IV
administration.
As in Fig 2 samples corresponding to each CD molarity are lyophilized and then put on stability at
25 C and 40 C for 30 days and 60 days in order to evaluate API:CD complexes stability in dry
phase.
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How to prove complex formation?

By Differential Scanning Calorimetry (DSC), we can see the API peak disappearing due to
API:-CDs complexation.

Fig. 5 DSC Thermograms of Lyophilized Zotepine Phase Solubility Samples

ZOTEPINE Phase Solubility Conclusions:

An AL type phase solubility was observed with all tested cyclodextrins
(1mol: 1mol of API:CD ratio).
A high complexation efficiency and stability constants were obtained for BCD and Crysmeb.
Compared to its solubility in water Zotepine solubility increased by ~2871, 1543 and 1240
times in presence of Crysmeb, HPBCD and SBE respectively.
The stability of the complexes formed in presence of Crysmeb>SBE>HPBCD>BCD following
30 and 60 days at 25 C and 40 C.
The results are offering to the formulator a wide choice in cyclodextrin selection for Zotepine
solubilization.
The complex formation is confirmed for both Crysmeb and HPBCD by DSC
Both Crysmeb and HPBCD would be ideal candidates for Zotepine solubilization.

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How can we solubilize an API by solid dispersion inclusion complexes?

By spray drying, freeze drying, kneading, physical mixture, etc. For all these procedures:
Find a volatile hydrophilic organic solvent in which your API is soluble
Prepare a saturated solution of the API in that organic solvent
Evaluate the ratio of the organic solvent (loaded with API) miscible with water (loaded with CD)
to get a clear solution (to avoid API precipitation out) in order to comply to AL complexation
profile (1mol: 1mol of API: CD ratio) as per the Phase solubility diagram ( Fig.3 ) for Zotepine.

How to get API:CDs complexation by Physical Mixture?

Simply mix in a V blender or Turbula mixer the two powders in the molar ratio as per phase
solubility. Depending on the API properties, the rate of encapsulation may be lower compared to
other techniques as Spray drying (SD), Freeze drying (Lyo), Physical mixture/kneading (PM).

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How to get API:CDs complexation by spray drying (solubilization from liquid to

solid state)?

How to get API:CDs complexation by freeze-drying (solubilization from liquid

to solid state)?

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How do I decide which method is the best to solubilize an API as a solid

dispersion?
The best method to create solid dispersions is decided based on DSC and dissolution assessment.

Fig.6 Thermograms for Zotepine Crysmeb Solid Dispersions

Fig.7 Thermograms for Zotepine HPBCD Solid Dispersions

Fig.8 Dissolution evaluation of Solid Dispersions of Zotepine in different cyclodextrins

ZOTEPINE Complexation by solid dispersion methods Conclusion:

Zotepine:HPBCD solid dispersion complexes prepared by spray drying can make this API a
very good candidate for tablet formulation with improved drug solubility and dissolution profile
The data obtained following DSC clearly demonstrate the amorphous nature and complexation
of the drug with cyclodextrin
The dissolution efficiency ranking is :
for HPBCD = Spray Drying (SD) > Lyophilization > Physical Mixture (PM)
for Crysmeb = Spray Drying (SD) > Physical Mixture (PM)
HPBCD is the best solution for Zotepine formulation in a solid dispersion

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Case Study 2: Zaleplon as Model Drug for Cyclodextrin Solubilization

Zaleplon is a hypnotic or sedative medication that induce sleep. It is used for
treating insomnia
It is a potent sedative or hypnotic, commercially available as a tablet at 5mg,
10mg and 20mg (Sonata)
Physical-chemical properties
- Mol. wt.: 305.34
- Log P: 1.53-2.0
BCS class II (Low Solubility and High Permeability)
Oral Bioavailability is 30% due to low solubility and extensive first pass
metabolism

Inclusion study of Zaleplon: phase solubility determination

Fig. 9 Phase-solubility profiles of Zaleplon in BCD,HPBCD, and Crysmeb at RT (25oC) in deionized water (pH 6.5).

Table 3. Zaleplon Solubility Enhancement at RT (25oC) in deionized water (pH 6.5).

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Complex formation analysis by Differential Scanning Calorimetry (DSC)

method

Fig. 10 DSC Thermograms of Lyophilized Zaleplon Phase Solubility Samples

The DSC chromatogram shows the peak disappearance and proves the API:BCDs complexation.

Complex characterization: stability constant and complexation efficiency

Table 4 Zaleplon complexes K 1:1 and CE at RT (25C) in delionized water (pH 6.5)

The complexation efficiency and stability constants are high when complexed with Crysmeb
and HPBCD

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Complex stability evaluation

Fig. 11 Zaleplon:CD complex stability evaluation at 25C and 40C.

The stability of the complexes after 30 and 60 days at 25C and 40C is ranking as follows:
Crysmeb > HPBCD > BCD SBE

ZALEPLON phase solubility Conclusion:

Zaleplon displays an AL type phase solubility profile in all tested cyclodextrins
The complexation efficiency and stability constants are high when complexed with Crysmeb
and HPBCD
Zaleplon solubility augmented by ~ 9, 5 and 3 times in presence of Crysmeb, HPBCD and
SBE respectively, compared to its solubility in water.
The stability of the complexes after 30 and 60 days at 25C and 40C is ranking as follows:
Crysmeb > HPBCD > BCD SBE
BCD solubilization potential is equal to SBE but is lower than HPBCD and Crysmeb.
As confirmed by DSC both HPBCD and Crysmeb are very good solubilization options
for improving Zaleplon solubility enhancement.

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Zaleplon solid dispersion inclusions prepared by: spray drying (SD), freeze
drying (LYO), physical mixture (PM=kneading)

Fig. 12 Thermograms for Zaleplon-Crysmeb Solid Dispersions

Fig. 13 Thermograms for Zaleplon-HPBCD Solid Dispersions

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Fig. 14 Dissolution evaluation of Solid Dispersions of Zaleplon in different Cyclodextrins

ZALEPLON complexation by solid dispersion methods Conclusion:

Solid dispersions of Zaleplon complexes with Crysmeb and HPBCD by spray drying are very
good candidates for drug solubility increase and improved dissolution profile which will result in
optimized solid dosage forms for its oral delivery
DSC confirms the change in crystalline nature of the drug by the complete disappearance of
Zaleplon peak in spray drying (SD) thermograms
For both CDs (Crysmeb and HPBCD) the dissolution efficiency ranking is:
Spray Drying (SD) > Physical Mixture (PM)
Due to the large amount of organic solvent needed for Zaleplon solubilization, the lyophilized
solid dispersions did not perform as expected.

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CONCLUSION
As demonstrated in the case studies of this document, -Cyclodextrins are fully
adapted tools to increase the aqueous solubility of APIs and their physico-chemical
stability.
They can also be used to reduce toxicity (e.g: local irritation after topical or oral
administration) and to mask material bad taste or odor.
Several pharmaceutical products containing Hydroxypropyl Betacyclodextrin have
already received market approval from several regulatory authorities and have
successfully reached the market.
The patent situation has evolved, in favour of more uses of HPBCD. This new
context makes this excipient an easily accessible formulation aid for the
development scientist.
Roquette provides a range of KLEPTOSE HPBCD derivatives suitable for the
cosmetics and the pharmaceutical industries, including a PYROGEN FREE grade
for parenteral applications. KLEPTOSE Crysmeb is a product under development
showing interesting inclusion properties.
Roquette is in a privileged position to discuss the specific needs of the
pharmaceutical and cosmetics industry for information on scientific assistance,
regulatory and patent issues.

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References (Roquettes case studies)

Determination of the Thermodynamic Solubility and the Affinity (Binding) Constants of Carbamazepine,
Danazol and Albendazole in Hydroxypropyl Beta Cyclodextrin (KLEPTOSEHPB) Solutions; Carmen Popescu,
Hassan Almoazen, Wenli Lu, Anthony Samsa , Leon Zhou, Ashish Joshi, James Johnson, AAPS 2011, Washington
Complexation of Furosemide with SBE--CD Studied by Phase Solubility and Solid Dispersion Methods; Rui
Zhu, Carmen Popescu, Jean-Yves Pierquin, Xia Zhao, Michael Clarke, Hassan Almoazen, Ed Brunson, James R
Johnson AAPS 2011, Washington
Effect of Preparation Method on Physical-Chemical
Properties and Dissolution Profile of Furosemide-Sufobutylether--cyclodextrin (SBE--CD) Solid Dispersions;
Rui Zhu, Carmen Popescu, Jean-Yves Pierquin, Xia Zhao, Michael Clarke, Hassan Almoazen, Ed Brunson, James R
Johnson, AAPS 2011,
Washington
The Influence of -Cyclodextrin Side Chain Substitutions on the Complexation Efficiency of a Model BCS
Class II Compound; C. Popescu, C. Wiley, l. Zhou, P. Lefevre, and L. Felton, AAPS 2011,Washington
Physicochemical Properties that Influence Cyclodextrin Complexation; C. Popescu , C. Wiley , P. Lefevre , A. J.
Hopfinger, L. Zhou1 , E.X. Esposito , and L. Felton ;AAPS 2012 Chicago
Lorazepam Complexation with Hydroxypropyl--cyclodextrin (HP--CD) and Sufobutylether--cyclodextrin
(SBE--CD): Phase Solubility Parameters Evaluation; Carmen Popescu , Wenli Lu, Leon Zhou , Hassan Almoazen ,
James Johns; AAPS 2012 Chicago
Influence of Cyclodextrin Complexation Method on the Physical Properties and Dissolution Profile of a BCS
Class II Model Drug; C. Popescu , C. Wiley , S. Potharaju , P. Lefevre , L. Zhou, D. Peterson, and L Felton; AAPS
2012 Chicago
Are Cyclodextrins a Viable Tool for Zotepine Solubilization? Prashanth Manda, Carmen Popescu, Abhishek
Juluri, Leon Zhou, Michael A. Repka, S. Narasimha Murthy,; AAPS 2013, San Antonio
Preparation And Charecterization Of Zotepine Solid Dispersions By Cyclodextrin Complexation; Carmen
Popescu, Prashanth Manda, Abhishek Juluri, Leon Zhou, Michael A. Repka, S. Narasimha Murthy; AAPS 2013, San
Antonio
Zaleplon (BCS Class II Model Drug) Solubility Enhancement By Native And Modified -Cyclodextrins; Carmen
Popescu, Prashanth Manda, Abhishek Juluri ,Leon Zhou , Michael A. Repka ,
S. Narasimha Murthy; AAPS 2013, SanAntonio
Preparation And Characterization Of Zotepine Solid Dispersions By Cyclodextrin Complexation; Carmen
Popescu ,Prashanth Manda, Abhishek Juluri, Leon Zhou, Michael A .Repka, S. Narasimha Murthy; AAPS 2013, San
Antonio
Solid Dispersion Of Zaleplon Cyclodextrin Complexes And Evaluation Of Their Efficiency; Prashanth Manda,
Carmen Popescu, Abhishek Juluri, Leon Zhou, Michael A. Repka, S. Narasimha Murthy; AAPS 2013, San Antonio
Celecoxib (BCS Class II Model Drug) Solubility Enhancement By Cyclodextrin Complexation; Abhishek Juluri,
Carmen Popescu, Prashanth Manda, Leon Zhou,Michael A. Repka , S. Narasimha
Murthy; AAPS 2013,San Antonio
Preparation And Charecterization Of Zaleplon Solid Dispersions By Cyclodextrin Complexation; Carmen
Popescu ,Prashanth Manda, Abhishek Juluri , Leon Zhou, Michael A .Repka , S. Narasimha
Murthy; AAPS 2013, San Antonio
Ampelopsin (Amp) Solubilization By Inclusion Complexes; Carmen Popescu, Abhishek Juluri, Craig Buske, Leon
Zhou, Philippe Lefevre, S. Narasimha Murthy; AAPS 2014, San Diego

Pharma.business.unit@roquette.com

www.roquettepharma.com

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