Hydrophobic Ion Pairing (HIP)

Encapsulating small molecules, peptides, and proteins

into nanocarriers

Silvia Argelia Peraza Ku

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Definition and nomenclature
Hydrophobic ion pairing (HIP) / Ionic complexation is the process of forming ionic interactions between a
charged hydrophilic molecule with an oppositely-charged counterion.

Hydrophobic counterion /
Ion pair / Ion pairing agent

Hydrophobic
complex / HIP
complex

Active pharmaceutical ingredient

(API) / Therapeuthic

The hydrophobic groups of the

The molecule’s charge is masked,
counter ion help to coat the original
mitigating solubility
molecule’s Surface area
Why use HIP?
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Challenge

However
Strategies for
HIP allows

Then
Low drug loadings, hydrophobic
encapsulation by
poor encapsulation compounds: emulsions,
increasing the
efficiencies, lack of nanoprecipitations,
hydrophobicity od
scalability. solid-lipid
therapeutics.
nanoparticles, etc.

Objective
To modify a drug’s solubility profile in order to make it more hydrophobic for encapsulation.
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HIP basics: solubility
The solubility of a species is determined by:
Entropy
State of disorder in a system. It is determined by the statistical numbers of configurations a system can attain.

or
or

Solvent Therapeutic

• For small molecules, that entropy is determined by the concentration of the solute in the solvent.
• Entropy always favors dissolution, that means, the degree of freedom in the system is favored.
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HIP basics: solubility
The solubility of a species is determined by:
Enthalpy
The sum of the internal energy (heat content) of a system.

We know that the entropy

∆H1 change for dissolution of a
solid is positive, so the
solubility depends on the
enthalpy change for the
overall process
Solvent

The assumption is made

that the more endothermic
(or less exothermic)
∆H2 the enthalpy of solution is,
Solution the less soluble the
compound.

Solute
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HIP basics: solubility
The solubility of a species is determined by:
The dielectric constant
Determines the strength of electrostatic interactions between Elementary charges.

As the dielectric constant

increases, therefore, the
𝒌 𝒒𝟏 𝒒𝟐
𝑭= 𝟐
force holding ions 𝜺𝒓
together decreases.
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Eligibility for HIP
Is my molecule suitable for HIP ?
For a therapeutic molecule to be eligible
for HIP:
• It must contain at least one charged
group.
• The molecule charge density (charge
per molecular weight) must be taken
Hydrophilic molecules Zwitterionic into consideration.

Commonly used counter ions

• Counter ions for hydrophobic ion
SO3
-
pairing should contain at least one
Phenyl SOO3- charged groups and at least one
(CH2)xCH3) PO(OH)O- hydrophobic domain.
COO-
• Ionic surfactants and sulfates are
+
R3N Phenyl common and popular.
+
H3N (CH2)xCH3)
• The most common cations are
quaternary amines and alkylamines.
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Key parameters to consider for HIP (choosing counterions)
1. Hidrophobicity

Log P: the logarithm of the octanol-water partition coefficient

Log P is the logarithm of the partition coefficient of a The most hydrophobic counterion is
solute between octanol and water and serves as a not always the best to use. Extremely
measure of the preference of the compound to hydrophobic counterions are difficult
dissolve in either water or an organic solvent. to dissolve in water for ion pairing.

1. pKa and pH

• Operating at a pH near one species’ pKa value is not advised.

• Curves denoting net charge vs. pH are available for many proteins in
the literature.
• Pinkerton et al. note that pKa values of the two charged species
should be different by at least two pH units for an ion to pair reliably
form.

Kakati et al., 2012

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Key parameters to consider for HIP
1. Complexation: pre-formed vs. in situ.
Pre-formed In-situ

Have a know stoichiometry and are already paired

together, meaning electrostatic interactions between the Less common
drug and the other delivery vehicle components are less Avoids micellization
likely to occur during encapsulation.

1. Charge ratio

At the higher charge ratios,

complex formation is not less
efficient than at the 1 : 1
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Encapsulation techniques
Considerations:
Encapsulation Efficiency (EE)

% 𝑬𝑬=𝟏𝟎𝟎 𝒙 𝟏 − (
𝒎𝒂𝒔𝒔 𝒐𝒇 𝒖𝒏𝒆𝒏𝒄𝒂𝒑𝒔𝒖𝒍𝒂𝒕𝒆𝒅 𝑨𝑷𝑰
𝒕𝒐𝒕𝒂𝒍 𝒎𝒂𝒔𝒔 𝒐𝒇 𝑨𝑷𝑰 )
Drug loading (DL)

% 𝑫𝑳=𝟏𝟎𝟎 𝒙 𝟏 − ( 𝒎𝒂𝒔𝒔 𝒐𝒇 𝒖𝒏𝒆𝒏𝒄𝒂𝒑𝒔𝒖𝒍𝒂𝒕𝒆𝒅 𝑨𝑷𝑰

𝒕𝒐𝒕𝒂𝒍 𝒎𝒂𝒔𝒔 𝒐𝒇 𝒗𝒆𝒉𝒊𝒄𝒍𝒆𝒔 )

Encapsulation techniques
Emulsions
There are two combinations
of emulsions:
• water/oil emulsions or
oil/water emulsions
• water/oil/water double
emulsions.
Precipitation
Based on precipitation mechanism.
Polymer precipitation occurs after
the addition of a non-solvent to a
polymer solution in four steps
mechanism: supersaturation,
nucleation, growth by condensation,
and growth by coagulation.
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Lipid nanoparticles
Lipid nanoparticles have emerged as an
alternative to polymer-based systems.
These are dispersions of submicron-
sized particles composed of a solid lipid
matrix that is solid at body and room
temperature, stabilized by a surfactant.
Others
Other formulation strategies can be viewed
through a similar lens to the one we have used
above.
• Techniques that treat a preformed complex
such as a typical hydrophobic molecule are
valid.
• Systems that require pH modulation are not
wholly ineligible for use with HIP.
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Ion paired drug release from a delivery vehicle

De-complexation is driven by one of two
main mechanisms:

• Counterion competition by salts

• pH-driven charge negation.

The former occurs when salts in the

surrounding medium can access the
complex and outcompete the hydrophobic
ion pair, leading to dissociation.
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Conclusion
Encapsulating hydrophilic therapeutics via hydrophobic ion pairing is a
useful technique and offers several attractive possibilities:
1. Co-encapsulating hydrophilic and hydrophobic drugs,

2. Forming a HIP complex from two therapeutic species,

3. Decreasing API crystallinity,

4. Tuning drug release rates by simply altering the counterion used in HIP.
Thank you!