biomolecules

Review
Calixarenes: Generalities and Their Role in
Improving the Solubility, Biocompatibility, Stability,
Bioavailability, Detection, and Transport
of Biomolecules
Edilma Sanabria Español 1, * and Mauricio Maldonado Villamil 2, *
1 Grupo GICRIM, Programa de Investigación Criminal, Universidad Manuela Beltrán, Avenida Circunvalar
No. 60-00, Bogotá 111321, Colombia
2 Departamento de Química, Facultad de Ciencias, Universidad Nacional de Colombia, Sede Bogotá,
Cr. 30 No. 45-03, Bogotá 111071, Colombia
* Correspondence: edilma.sanabria@docentes.umb.edu.co (E.S.E.); mmaldonadov@unal.edu.co (M.M.V.);
Tel.: +57-1-5460600 (ext. 1153) (E.S.E.); +57-1-3165000 (ext. 14414) (M.M.V.)




Received: 25 January 2019; Accepted: 28 February 2019; Published: 5 March 2019


Abstract: The properties and characteristics of calix[n]arenes are described, as well as their capacity to
form amphiphilic assemblies by means of the design of synthetic macrocycles with a hydrophilic head
and a hydrophobic tail. Their interaction with various substances of interest in pharmacy, engineering,
and medicine is also described. In particular, the role of the calix[n]arenes in the detection of
dopamine, the design of vesicles and liposomes employed in the manufacture of systems of controlled
release drugs used in the treatment of cancer, and their role in improving the solubility of testosterone
and anthelmintic drugs and the biocompatibility of biomaterials useful for the manufacture of
synthetic organs is emphasized. The versatility of these macrocycles, able to vary in size, shape,
functional groups, and hydrophobicity and to recognize various biomolecules and molecules with
biological activity without causing cytotoxicity is highlighted.

Keywords: calixarenes; complexion; amphiphilic assemblies; micelles; vesicles; liposomes; nanoparticles;

encapsulation; transport of drugs; bioavailability; solubility of the drugs

1. Introduction
In the fields of pharmacy, medicine, and nutrition, there are several macrocycle molecules that
have been used for improving the solubility of the active ingredient of drugs and their stability and
bioavailability for the determination of proteins and nucleic acids. They have even exhibited biological
activity or value in sensitization therapies, as seen for example with cucurbit[n]urils [1], macrocyclic
tetrapyrazolics [2], resorcinarenes [3], crown ethers [4], cyclodextrins [5], phthalocyanines [6],
and calixarenes [7], among others. All of this has been possible due to the fact that these compounds are
able to form a complex with molecules of biological interest such as antibiotics [1], proteins and nucleic
acids [4], food nutraceuticals [5], and cholesterol [7], among many other compounds. In particular,
the calixarenes are able to form a guest–host inclusion complex with molecules such as anthelmintics [8],
testosterone [9], steroid hormones [10], antibacterial drugs [11], sugars [12], alditols [13], lectin [14],
protein [15], and pesticides [16,17]. In addition, they have exhibited antiviral [18], antithrombotic [19],
antibactericidal [20], antituberculosis [21], and anticancer activity [22].
In this article, the characteristics and properties that enable the calixarenes to self-assemble in
order to form amphiphilic assemblies, micelles of cylindrical or ellipsoidal geometry, vesicles, bilayers,
liposomes, and nanoparticles such as nanocapsules and nanospheres that allow the encapsulation,

Biomolecules 2019, 9, 90; doi:10.3390/biom9030090 www.mdpi.com/journal/biomolecules

Biomolecules 2019, 9, 90 2 of 15

transport, and9,controlled
Biomolecules 2019, release of drugs or that can play an important role in the stability,
x FOR PEER REVIEW 2 of 17
bioavailability, solubility, or even the biological activity of the drugs are reviewed. Also reviewed are
stability,
the types ofbioavailability,
molecules that solubility, or even thebybiological
can be recognized activity
calixarenes, theirof the drugs are
interactions, andreviewed. Also
the techniques
reviewed are the types of molecules that can be recognized by calixarenes, their interactions,
that allow their determination and the evaluation of the stoichiometry of the complex formed and thein
techniques that allow their determination and the evaluation of the stoichiometry of the complex
solution and solid state. In particular, calixarene toxicity, their characteristics, the variation in the size
offormed in solution and solid state. In particular, calixarene toxicity, their characteristics, the variation
the cavity and in their functional groups, and their applications, as well as the advantages of these
in the size of the cavity and in their functional groups, and their applications, as well as the
macrocycles, are described.
advantages of these macrocycles, are described.
2. Calix[n]arenes
2. Calix[n]arenes
2.1. Summary Description of Its Structure
2.1. Summary Description of Its Structure
Polyhydroxylated platforms such as calixarenes, resorcinarenes, or pyrogalloarenes (Figure 1)
Polyhydroxylated
are a very platforms
interesting class such as calixarenes,
of compounds. They haveresorcinarenes, or pyrogalloarenes
a remarkable ability (Figurefor
to act as receptors 1)a
are a very
variety interesting
of guest species,class of compounds.
depending on theirThey have properties
structural a remarkable abilitywhich
[23–27], to actcanas receptors
be modifiedfor by
a
variety of guest species, depending on their structural properties [23–27], which can be
changing the size of the substituents or by adding functional groups as a part of the scaffold [28–32].modified by
changing
Among thethedescribed
size of themacrocyclic
substituentscompounds,
or by adding calixarenes
functional groups as a partthe
are probably of the scaffold
most [28–32].
promising for
Among the described macrocyclic compounds, calixarenes are probably the most promising
application in the area of host–guest recognition of toxicological molecules. Their synthetic availability, for
application
low toxicity, inandthepresence
area ofofhost–guest recognition
reactive sides of toxicological
are characteristics molecules.
that make them Their synthetic
relevant within
availability, low toxicity,
supramolecular chemistry. and presence of reactive sides are characteristics that make them relevant
within supramolecular chemistry.
OH R HO OH R HO
R R HO OH

HO OH HO OH
OH HO
R R R R
OH HO
HO OH HO OH

R R HO OH
OH R HO OH R HO

a b c
Figure 1. Polyhydroxylated plataforms. (a) Calix[4]arene; (b) resorcinarene; (c) pyrogalloarene.
Figure 1. Polyhydroxylated plataforms. (a) Calix[4]arene; (b) resorcinarene; (c) pyrogalloarene.
The calixarenes are a family of macrocyclic compounds with a variable number of phenol units
The calixarenes are a family of macrocyclic compounds with a variable number of phenol units
linked by methylene bridges in ortho position [33]. The number of units of aromatics can be between
linked by methylene bridges in ortho position [33]. The number of units of aromatics can be between
4 and 20, although the calixarenes of 4, 5, 6, 7, and 8 are the most common [34,35]. As shown in
4 and 20, although the calixarenes of 4, 5, 6, 7, and 8 are the most common [34,35]. As shown in Figure
Figure 2, the cyclic structure of calix[n]arenes is similar to other polyhydroxylated macrocyles [36,37],
2, the cyclic structure of calix[n]arenes is similar to other polyhydroxylated macrocyles [36,37], and
and the macrocycle cavity will depend on the number of aromatic units in the system. These
the macrocycle cavity will depend on the number of aromatic units in the system. These compounds
compounds have a three-dimensional cavity that can accommodate host molecules during a process
have a three-dimensional cavity that can accommodate host molecules during a process called host–
called host–guest complexation. These systems have an advantage as a synthetic receptor, owing to
guest complexation. These systems have an advantage as a synthetic receptor, owing to different
different conformational isomeric forms, which allow different uses and applications. For instance,
conformational isomeric forms, which allow different uses and applications. For instance, the
the calix[4]arenes can adopt several different conformers, including the cone, partial cone, 1,2-alternate,
calix[4]arenes can adopt several different conformers, including the cone, partial cone, 1,2-alternate,
and
and1,3-alternate.
1,3-alternate. In
In the
the rigid
rigid cone
cone conformation, all the
conformation, all the phenolic
phenolic –OH
–OHgroups
groupsform
formstrong
stronghydrogen
hydrogen
bonds that stabilize the structure (Figure
bonds that stabilize the structure (Figure 2).2).
Biomolecules 2019, 9, 90 3 of 15
Biomolecules 2019, 9, x FOR PEER REVIEW 3 of 17

R R R R

HO
OH HO OH HO
OH HO
OH HO
OH

R R
R R

R
I. Calix[4]arene
R II. Calix[6]arene

R R

HO
OH HO

R OH HO R

OH HO
OH

R R

R
III. Calix[8]arene

R R R R R R R R R R R
OH OH
OH
OH
OH
OH OH OH HO OH OH HO OH OH OH OH

R R
R
R
R

IV. Cone V. Partial Cone VI. 1,2-alternate VII. 1,3-alternate

Figure 2. Structures of calix[4]arenes.

Figure 2. Structures of calix[4]arenes.

In the cavity of calix[4]arene in the cone conformation, it is possible to distinguish two edges: the
lower rim, Inwhere the of
the cavity methylene bridges
calix[4]arene in theare,
coneand the upper rim
conformation, on the opposite
it is possible side. two
to distinguish In the center is
edges:
the annular
the lower system, where
rim, where the the aromatic
methylene ringsare,
bridges are;
andinthe
addition,
upper rimtheonupper rim is larger
the opposite side. Inthan the lower
the center
rim (Figure 3) [38].
is the annular The molecular
system, dimensions
where the aromatic ringsof the
are; incavities
addition,vary depending
the upper on the
rim is larger number
than of units
the lower
rim (Figure
of aromatics; 3) [38].
thus, the The molecular
diameter dimensions
of the upper rim of has
the cavities vary depending
been estimated onÅ
to be 3.8 the number
for of units and
calix[4]arenes
Biomolecules
thus,2019, 9, x FOR PEER REVIEW 4 of 17
5.0 Åoffor
aromatics;
calix[6]arenes the diameter
[39]. of the upper rim has been estimated to be 3.8 Å for calix[4]arenes and
5.0 Å for calix[6]arenes [39].

Upper rim

HOH
O
OH HO
OH
OH HO
OH Lower rim

Figure 3. Upper rim and the lower rim on the cavity of the calix[4]arenes.
Figure 3. Upper rim and the lower rim on the cavity of the calix[4]arenes .

The calix[4]arenes can be functionalized on the upper rim or on the lower rim with several
functional groups such as amides, imines, sulfur, azo, semicarbazone, and alkyl groups, among
others, producing a wide variety of macrocycle compounds with different properties of recognition,
selectivity, solubility, and degree of hydrophobicity [40]. This last aspect is very important, because
with the introduction of polar groups to the calixarenes it is possible to design amphiphilic
Biomolecules 2019, 9, 90 4 of 15

The calix[4]arenes can be functionalized on the upper rim or on the lower rim with several
functional groups such as amides, imines, sulfur, azo, semicarbazone, and alkyl groups, among others,
producing a wide variety of macrocycle compounds with different properties of recognition, selectivity,
solubility, and degree of hydrophobicity [40]. This last aspect is very important, because with the
introduction of polar groups to the calixarenes it is possible to design amphiphilic macrocycles with a
hydrophilic head and a hydrophobic tail, which may self-assemble into micelles, vesicles, liposomes,
and other aggregates useful in the transport of drugs [19]. Functionalization of calixarenes can be
done from the starting materials; varying the nature of the substituent group on the phenol facilitates
the modification on the upper rim of the macrocyclic system. On the lower rim, the obvious site for
chemical modification is that of the hydroxyl groups. The functionalization of calixarenes with polar
groups can lead to various structures, as shown in Figure 4. The reactivity of the calixarenes is mainly
located at two points: on the hydroxyl groups (lower rim) or on the position of the hydroxyl group
(upper rim). Functioning substances such as carboxilates [41], phosphates [42], ammonium groups [43]
or sulfonates [44] can be introduced into the hydroxylated platform by means of easily accessible
reactions and
Biomolecules 2019,selectively, with
9, x FOR PEER good yields.
REVIEW 5 of 17

O R 4
O O

HO O OH

OH 4
OH 4

R R HO
ONa O
R R P
O S O
OH

OH 4 OH HO OH 4
OH
HO
Calix[n]arene

NH2
R

OH 4 HO
O 4
O
OH P
S O
OH
O

OH 4

Figure 4. Polar derivatives of calixarenes.

calix[n]arenes lies in the fact that they are able to recognize and
The importance of the calix[n]arenes
accommodate into
accommodate intotheir cavity
their guestguest
cavity molecules via non-covalent
molecules interactions.
via non-covalent Some of these
interactions. Someinteractions
of these
interactions are H-bonding, cation–π, π–π stacking, van der Waals interactions, and the so-called
anion–π, in cases where the aromatic system is electron-deficient [45,46]. The magnitude of the
interaction also depends on the conformation of the macrocycle [47].
In conclusion, the advantages of the calix[4]arenes are that they are easy to synthetize and that
they can be modified to obtain compounds according to the guest that one wants to complex. This
Biomolecules 2019, 9, 90 5 of 15

are H-bonding, cation–π, π–π stacking, van der Waals interactions, and the so-called anion–π, in cases
where the aromatic system is electron-deficient [45,46]. The magnitude of the interaction also depends
on the conformation of the macrocycle [47].
In conclusion, the advantages of the calix[4]arenes are that they are easy to synthetize and that they
can be modified to obtain compounds according to the guest that one wants to complex. This synthesis
has good yields, and the reagents are inexpensive [48]. In addition, the simple derivatives of calixarenes
have not exhibited toxicity or immunogenic properties [19]. All of this makes calixarenes highly valued
in supramolecular chemistry as complex agents for the transport of drugs and their controlled release,
among other applications.

2.2. Complexing Properties of Calixarenes

As indicated, the cavity of the calixarenes is of variable size and is sufficiently large to
accommodate anions, cations, or neutral molecules, including biologically important molecules. This,
together with their ability to trap guest molecules by means of noncovalent interactions, has resulted
in widespread interest in the calix[n]arenes within supramolecular chemistry, particularly with respect
toBiomolecules 2019, 9,phenomena
the host–guest x FOR PEER REVIEW
(Figure 5) [49,50]. 6 of 17

Figure 5. Three-dimensional representation of the complexation process of calix[4]arenes.

Figure 5. Three-dimensional representation of the complexation process of calix[4]arenes.
Furthermore, as mentioned above, the calix[n]arenes can be functionalized at both rims, R and
–OH (Figure 3) [51–53],
Furthermore, producing aabove,
as mentioned wide the
variety of compounds
calix[n]arenes that
can be can have different
functionalized at bothcomplexing
rims, R and
properties. In particular, calix[n]arenes functionalized with a sulfonic group in the para
–OH (Figure 3) [51–53], producing a wide variety of compounds that can have different complexing position have
been found to have several pharmaceutical applications, due to their potential for encapsulating
properties. In particular, calix[n]arenes functionalized with a sulfonic group in the para position have drugs,
increasing
been foundtheirtosolubility,
have severalbioavailability, oral absorption,
pharmaceutical and
applications, stability
due to theirunder heat, light,
potential and acidic
for encapsulating
conditions [54–56]. In addition, they possess good biocompatibility and innocuousness [56,57].
drugs, increasing their solubility, bioavailability, oral absorption, and stability under heat, light, Other
and
applications of the calix[n]arenes are as phase-transfer agents, sensors, antibacterials,
acidic conditions [54–56]. In addition, they possess good biocompatibility and innocuousness [56,57].ion-selective
electrodes, and use as of
Other applications catalysts and in separations
the calix[n]arenes are as science [58,59]. agents, sensors, antibacterials, ion-
phase-transfer
selective electrodes, and use as catalysts and in separations sciencesystems
Other advantages of the calix[n]arenes over other macrocyclic [58,59]. include a preorganized
cavity Other
of variable size and
advantages an electron-rich
of the calix[n]arenesoption of modification
over other macrocyclicand selective
systems binding
include with the
a preorganized
guest [58,59]. This increases their potential use as agents of specific recognition, for example
cavity of variable size and an electron-rich option of modification and selective binding with the of a guest
guest
of[58,59].
toxicological interest.
This increases their potential use as agents of specific recognition, for example of a guest of
toxicological interest.
2.3. Amphiphilic Assemblies Based on Calixarenes
2.3.As
Amphiphilic Assemblies
was discussed Basedcalixarenes
above, on Calixarenes
are substances of easy access and of easy chemical
modification, thus allowing one to obtain systems with the introduction of polar groups to the
As was discussed above, calixarenes are substances of easy access and of easy chemical
calixarenes, making it possible to design amphiphilic macrocycles with a hydrophilic head and a
modification, thus allowing one to obtain systems with the introduction of polar groups to the
hydrophobic tail, which may self-assemble into micelles, vesicles, liposomes, nanocapsules, and other
calixarenes, making it possible to design amphiphilic macrocycles with a hydrophilic head and a
aggregates. The development of self-organizing synthetic amphiphilic calixarenes with properties of
hydrophobic tail, which may self-assemble into micelles, vesicles, liposomes, nanocapsules, and other
aggregates. The development of self-organizing synthetic amphiphilic calixarenes with properties of
inclusion and encapsulation allows complex hydrophobic molecules to be transported in a
hydrophilic environment; this is especially useful in the transport of drugs (Figure 6).
Biomolecules 2019, 9, 90 6 of 15

inclusion and encapsulation allows complex hydrophobic molecules to be transported in a hydrophilic

environment;
Biomolecules 2019,this is especially
9, x FOR useful in the transport of drugs (Figure 6).
PEER REVIEW 7 of 17

Bilayer

Nanocapsule Micella

OH
HO O
HO O
O
P O OH P O
P
O
O OH P HO
OH O
HO

Inverted micella Vesicule

Figure 6. Possibilities of amphiphilic calixarenes in self-assembly.

2.4. Types of MoleculesFigure

that Can Be Recognized
6. Possibilities by Calixarenes
of amphiphilic and Their
calixarenes Effect
in self-assembly.
As mentioned above, the calixarenes have potential applications in diverse fields of medicine and
2.4.the
in Types of Molecules that
pharmaceutical and Can Be Recognized
biological contexts.byThey
Calixarenes
are veryand Their Effect
versatile molecules, because they have a
hydrophobic character, but when aqueous environments are needed,
As mentioned above, the calixarenes have potential applications in for example
diverseinfields
biological media,
of medicine
the
andcalixarenes can be modified
in the pharmaceutical and by the introduction
biological contexts.of hydrophilic
They are verygroups at the
versatile upper and
molecules, lowerthey
because rim
of the macrocycle, leading to water-soluble structures. In this section, we give a brief
have a hydrophobic character, but when aqueous environments are needed, for example in biological description of
both
media, hydrophilic and hydrophobic
the calixarenes molecules
can be modified by thethat can be recognized
introduction or transported
of hydrophilic groups at bythe
calixarenes.
upper and
lower rim of the macrocycle, leading to water-soluble structures. In this section, we give a brief
2.4.1. Recognition of Dopamine by Calixarenes
description of both hydrophilic and hydrophobic molecules that can be recognized or transported by
calixarenes.
Among the many substances that can be recognized by calixarenes is dopamine (Figure 7), which
is a neurotransmitter of vital importance for the normal function of the central nervous system. The lack
2.4.1.
of thisRecognition
substance isofone
Dopamine by Calixarenes
of the causes of Parkinson’s disease, an illness that affects more than 10 million
people worldwide [60]. In addition,
Among the many substances that alterations in the normal
can be recognized levels of dopamine
by calixarenes is dopamine are(Figure
associated with
7), which
attention-deficit hyperactivity
is a neurotransmitter disorder, which
of vital importance for the affects
normalbetween
function2ofand
the 7% of children,
central nervous adolescents,
system. The
and adults in the world [61].
lack of this substance is one of the causes of Parkinson’s disease, an illness that affects more than 10
million people worldwide [60]. In addition, alterations in the normal levels of dopamine are
associated with attention-deficit hyperactivity disorder, which affects between 2 and 7% of children,
adolescents, and adults in the world [61].
Biomolecules 2019, 9, 90 7 of 15
Biomolecules2019,
Biomolecules 2019,9,9,xxFOR
FORPEER
PEERREVIEW
REVIEW 88ofof17
17

HHO
O NH22
NH

HHO
O
Figure 7. Chemical structure of dopamine.
Figure7.7.Chemical
Figure Chemicalstructure
structureofofdopamine.
dopamine.
Dopamine can be recognized by 4-tert-butylcalix[6]arene when it is co-spread with cellulose
acetate Dopamine
Dopamine
to form acan can berecognized
be
Langmuir recognized
film andby by 4-tert-butylcalix[6]arene
4-tert-butylcalix[6]arene
transferred to a gold electrode. when
when Theititcellulose
isisco-spread
co-spread with
with
acetate cellulose
cellulose
maintaining
acetate
acetate
the totoform
calixareneformremains
aaLangmuir
Langmuir film
in thefilm andtransferred
and
vertical transferred
cone totoaagold
conformation gold electrode.
inelectrode.
the Theinterface,
The
air-water cellulosewhile
cellulose acetate
acetatethe maintaining
maintaining
presence of
the
the calixarene
calixarene remains
remains in
in the
the vertical
vertical cone
cone conformation
conformation inin the
the air-water
air-water
calixarene in the Langmuir film provides selectivity to a sensor. The lineal range where the dopamine interface,
interface, while
while the
the presence
presence
ofof calixarene
can calixarene
be detected inin the
isthe Langmuir
Langmuir
between 5 and film
film provides
provides
100, selectivity
and 100selectivity
and 7500 to to aaand
nm, sensor.
sensor.
the The The lineal
limit lineal
of range where
range
detection iswhere the
2.54 the
nM.
dopamine
dopamine can
can be
be detected
detected isis between
between 55 and
and 100,
100, and
and 100
100 and
and 7500
7500 nm,
nm,
The ratio of calixarene/cellulose acetate has been optimized to show that when the percent of calixarene and
and the
the limit
limit of
of detection
detection isis
2.54
2.54 nM.
nM. The
The ratio
ratio of
of calixarene/cellulose
calixarene/cellulose
is 30 wt %, the reuptake of the dopamine is greatest [62]. acetate
acetate has
has been
been optimized
optimized toto show
show that
that when
when the
the percent
percent
ofofcalixarene
calixareneisis30 30wt wt%,%,the
thereuptake
reuptakeofofthe thedopamine
dopamineisisgreatest
greatest[62].
[62].
2.4.2. Improved Solubility of Testosterone with Calixarene
2.4.2.Improved
2.4.2. ImprovedSolubility
SolubilityofofTestosterone
TestosteronewithwithCalixarene
Calixarene
The interaction of calixarenes with steroids has been studied by several authors. In particular,
a study Theof
The interaction
interaction ofcalixarenes
of
the complexation calixarenes withsteroids
with
of testosteronesteroids
with has
has beenstudied
a been studiedby
water-soluble by severalauthors.
several
calixarene authors.
has been Inpublished
In particular,by
particular, aa
study
study of
of the
the complexation
complexation of
of testosterone
testosterone with
with aa water-soluble
water-soluble calixarene
calixarene
Millership [63]. Testosterone (Figure 8) is a steroid hormone involved in the male sexual response. has
has been
been published
published by
by
Millership
Millership
It is recognized[63].Testosterone
[63]. Testosterone (Figure8)8)isisaacan
that erectile(Figure
dysfunction steroid
steroid
result hormone
hormone
from low involved
involved
levels in inthe
of themale
malesexual
testosterone,sexual
and response.
response. ItIt
therefore
isrecognized
istherecognized
assessment that
that erectiletestosterone
oferectile
serum dysfunctioncan
dysfunction canresult
levelsresult
before from
from lowlevels
low levelsaof
establishing ofpossible
testosterone,
testosterone, andtherefore
and
treatment therefore
is important the
the
assessment
assessment
(Figure ofofserum
2) [64]. serumtestosterone
testosteronelevels
levelsbefore
beforeestablishing
establishingaapossible
possibletreatment
treatmentisisimportant
important(Figure
(Figure2)2)
[64].
[64].
OH
OH

HH

OO
Figure 8. Chemical structure of testosterone.
Figure8.8.Chemical
Figure Chemicalstructure
structureofoftestosterone.
testosterone.
Because testosterone is not soluble in water, a method that allows its solubilization is desirable.
Becausetestosterone
Because testosteroneisisnot
notsoluble
solublein inwater,
water,aamethod
methodthat thatallows
allowsitsitssolubilization
solubilizationisisdesirable.
desirable.
In this way, Millership improved the solubility of this steroid in water by means of the complexation
Inthis
In thisway,
way,Millership
Millershipimproved
improvedthe thesolubility
solubilityofofthis
thissteroid
steroidininwater
waterby bymeans
meansof ofthe
thecomplexation
complexation
of the testosterone with 4-sulphonic calix[n]arenes. The author measured the solubility in the presence
of the
of the testosterone
testosterone with
with 4-sulphonic
4-sulphonic calix[n]arenes.
calix[n]arenes. The The author
author measured
measured the the solubility
solubility inin the
the
and absence of several calixarenes, such as 4-sulphonic calix[4]arenes, 4-sulphonic calix[6]arenes,
presence and
presence and absence
absence ofof several
several calixarenes,
calixarenes, suchsuch asas 4-sulphonic
4-sulphonic calix[4]arenes,
calix[4]arenes, 4-sulphonic
4-sulphonic
and 4-sulphonic calix[8]arenes below the critical micelle concentration, and in all cases found the
calix[6]arenes,and
calix[6]arenes, and4-sulphonic
4-sulphoniccalix[8]arenes
calix[8]arenesbelowbelowthe thecritical
criticalmicelle
micelleconcentration,
concentration,and andininall
allcases
cases
formation of soluble complexes 1:1, with 4-sulphonic calix[8]arene being the most soluble. This was
foundthe
found theformation
formationof ofsoluble
solublecomplexes
complexes1:1, 1:1,with
with4-sulphonic
4-sulphoniccalix[8]arene
calix[8]arenebeing
beingthethemost
mostsoluble.
soluble.
explained by the fact that the steroid does not enter into the cavity of 4-sulphonic calix[4]arenes,
This was
This was explained
explained by by the
the fact
fact that
that the
the steroid
steroid does
does not
not enter
enter into
into the
the cavity
cavity of
of 4-sulphonic
4-sulphonic
due to their small size, while it can enter comfortably into the cavity of 4-sulphonic calix[6]arenes
calix[4]arenes,due
calix[4]arenes, duetototheir
theirsmall
smallsize,
size,while
whileititcan
canenter
entercomfortably
comfortablyinto intothe
thecavity
cavityofof4-sulphonic
4-sulphonic
and 4-sulphonic calix[8]arene. He also found that the pH of the solution influences the shape of
calix[6]arenesand
calix[6]arenes and4-sulphonic
4-sulphoniccalix[8]arene.
calix[8]arene.He Healso
alsofound
foundthat
thatthe
thepHpHofofthe
thesolution
solutioninfluences
influencesthe the
the calixarene because of the effect of the hydrogen bonding of the hydroxyl groups. Experiments
shape of
shape of the
the calixarene
calixarene because
because of of the
the effect
effect of
of the
the hydrogen
hydrogen bonding
bonding of of the
the hydroxyl
hydroxyl groups.
groups.
conducted in phosphate buffers at pH 5.8, 7.3, and 10 demonstrate that the greatest solubility is reached
Experimentsconducted
Experiments conductedin inphosphate
phosphatebuffersbuffersatatpHpH5.8,5.8,7.3,
7.3,and
and10 10demonstrate
demonstratethat thatthe
thegreatest
greatest
at pH 7.3, which suggests that the conformations adopted by calixarene at this pH favor the process of
solubilityisisreached
solubility reachedatatpHpH7.3,
7.3,which
whichsuggests
suggeststhat
thatthetheconformations
conformationsadopted
adoptedby bycalixarene
calixareneatatthis
this
complexation. The complex constants at pH 7.3 for complexes testosterone–4-sulphonic calix[4]arenes,
pHfavor
pH favorthe
theprocess
processofofcomplexation.
complexation.The Thecomplex
complexconstants
constantsatatpH
pH7.37.3for
forcomplexes
complexestestosterone–
testosterone–
testosterone–4-sulphonic calix[6]arenes, and testosterone–4-sulphonic calix[8]arenes were 26± 22,
4-sulphonic calix[4]arenes,
4-sulphonic calix[4]arenes, testosterone–4-sulphonic
testosterone–4-sulphonic calix[6]arenes,
calix[6]arenes, andand testosterone–4-sulphonic
testosterone–4-sulphonic
346 ± 39, and 156 ± 9, respectively [65].
calix[8]areneswere
calix[8]arenes were26±
26±22,
22,346
346±±39,
39,and
and156156±±9,9,respectively
respectively[65].
[65].

2.4.3.Biomaterial
2.4.3. BiomaterialModification
Modificationwith
withCalixarenes
CalixarenestotoAvoid
AvoidAllergy
AllergyororInfection
Infection
Thereare
There areseveral
severalimplantable
implantablemedical
medicaldevices
devicesuseful
usefulfor
forthetherepair
repairof
ofsoft
softand
andhard
hardtissue,
tissue,but
but
many of
many of these
these produce
produce acute
acute inflammation.
inflammation. Charnley
Charnley etet al.
al. [65]
[65] proposed
proposed coating
coating these
these medical
medical
Biomolecules 2019, 9, 90 8 of 15

2.4.3. Biomaterial Modification with Calixarenes to Avoid Allergy or Infection

There are several implantable medical devices useful for the repair of soft and hard tissue, but
many of these
Biomolecules 2019,produce acuteREVIEW
9, x FOR PEER inflammation. Charnley et al. [65] proposed coating these medical devices 9 of 17
with a natural anti-inflammatory that consists of a hormone produced by the organism that does
devices
not with allergy
generate a natural oranti-inflammatory
infection but that canthat easily
consists
beof a hormonein
synthesized produced by the organism
the laboratory. that
In this study,
does
the not generate
hormone allergy or infection but that
was α-melanocyte-stimulating can easily
(Figure 9), andbewas
synthesized in the
immobilized laboratory.
onto In this
medical device
study, the hormone was α-melanocyte-stimulating (Figure 9), and was immobilized onto
surfaces with C-tetra(octyl)calixresorcin[4]arene. The results obtained by the authors indicated that the medical
device surfaces
α-melanocyte with C-tetra(octyl)calixresorcin[4]arene.
hormone The results
retains its anti-inflammatory properties obtained by
and suggested thethis
that authors indicated
strategy could
that
be the α-melanocyte
useful in the manufacture hormone
of new retains its anti-inflammatory
materials [66]. properties and suggested that this
strategy could be useful in the manufacture of new materials [66].
O
OH H2N O
NH
NH O
HN
O OH O
NH OH O
NH O
N
O H2N O
NH
OH NH O O NH
S O
NH
NH O HN
N O O
NH
NH
NH
HN
NH2
N
H

Figure 9. Chemical
Figure 9. Chemical structure
structure of α-melanocyte hormone.
of α-melanocyte hormone.

Other applications similar to this one have been reported with proteins, which are important
Other applications similar to this one have been reported with proteins, which are important for
for the manufacture of synthetic organs, drug delivery systems, and biosensing, among other
the manufacture of synthetic organs, drug delivery systems, and biosensing, among other uses. The
uses. The aim is to achieve an efficient interaction between the proteins and the solid materials
aim is to achieve an efficient interaction between the proteins and the solid materials for the
for the production of nano- and biomaterials. In this context, the calixarenes have been shown
production of nano- and biomaterials. In this context, the calixarenes have been shown to be useful
to be useful as linking agents. For example, Keskinates et al. [66] used a calixarene tetraester
as linking agents. For example, Keskinates et al. [66] used a calixarene tetraester (5,11,17,23-tetra-tert-
(5,11,17,23-tetra-tert-butyl-25,26,27,28-tetramethoxycarbonylmethoxy-calix[4]arene) to achieve a good
butyl-25,26,27,28-tetramethoxycarbonylmethoxy-calix[4]arene) to achieve a good interaction
interaction between nanofibers of polyacrylonitrile (PAN) or poly(methyl methacrylate) (PMMA) and a
between nanofibers of polyacrylonitrile (PAN) or poly(methyl methacrylate) (PMMA) and a green
green fluorescent protein. The containment of calixarene on the fiber where the binding of protein was
fluorescent protein. The containment of calixarene on the fiber where the binding of protein was
highest was 50%. The usefulness of the modified fiber is as an adsorbent membrane for the removal
highest was 50%. The usefulness of the modified fiber is as an adsorbent membrane for the removal
of proteins in aqueous solution; therefore the authors suggest that PAN or PMMA fibers containing
of proteins in aqueous solution; therefore the authors suggest that PAN or PMMA fibers containing
calixarene are a promising new and inexpensive material for protein purification [67].
calixarene are a promising new and inexpensive material for protein purification [67].
2.4.4. Combination of Calixarenes and Cyclodextrins to Improve the Solubility of an
2.4.4. Combination
Anthelmintic Drugof Calixarenes and Cyclodextrins to Improve the Solubility of an Anthelmintic Drug
Niclosamide(Figure
Niclosamide (Figure10) 10) is
is an
an oral
oral anthelmintic
anthelmintic drug
drug used
used worldwide
worldwide to to treat
treat parasitic
parasitic infections.
infections.
It is active against beef and dog tapeworms. In addition, it is useful for
It is active against beef and dog tapeworms. In addition, it is useful for treating diseases treating diseases caused
caused by
by
these parasites,
these parasites, suchsuch as cancer,
as cancer, metabolic
metabolic diseases,diseases, artery constriction,
artery constriction, endometriosis,
endometriosis, and rheumatoid and
rheumatoid arthritis, among others [68]. In spite of its broad clinical application,
arthritis, among others [68]. In spite of its broad clinical application, the efficacy of niclosamide the efficacy of
niclosamide can be affected by its very low solubility in water (230 ng/mL).
can be affected by its very low solubility in water (230 ng/mL). Studies conducted by Yang et Studies conducted by
Yang
al. haveet shown
al. have shown
a great a great
increase increase
in the in the
aqueous aqueous
solubility solubility by
niclosamide niclosamide by means of
means of complexation
complexation with 4-sulphonate-calix[6]arene and hydroxypropyl-β-cyclodextrin.
with 4-sulphonate-calix[6]arene and hydroxypropyl-β-cyclodextrin. The advantage of The advantage
the use ofof
the use of water-soluble
water-soluble calixarenescalixarenes
is that they is that they
provide provide a hydrophobic
a hydrophobic environment environment
to include theto drug
include
andthe
a
drug and a hydrophilic head (sulphonate groups) that allow its solubilization.
hydrophilic head (sulphonate groups) that allow its solubilization. The combination of the two The combination of
the two macrocycle
macrocycle types provides
types provides both the both the properties
properties of cyclodextrins
of cyclodextrins andand those
those of of micelles.Another
micelles. Another
advantage is
advantage is that
that the
thep-sulphonate-calix[4,
p-sulphonate-calix[4,6,6,and and8]arenes
8]arenesdodo notnot
exhibit acute
exhibit toxication
acute whenwhen
toxication they
are injected into mice, and exhibit no activity in the Ames test. In addition, the inner cavity diameter
is variable, between 3.0, 7.6, and 11.7 Å for calix[4]arene, calix[6]arene, and calix[8]arene, respectively,
and between 5.7, 7.8, and 9.5 Å for α, β, and γ-cyclodextrins. Furthermore, calixarenes are highly
flexible molecules, while the cyclodextrins are quite rigid molecules. All of the above ensures good
versatility of the binding of the two macrocycles with the anthelmintic drug.
 Biomolecules 2019, 9, 90 9 of 15

they are injected into mice, and exhibit no activity in the Ames test. In addition, the inner cavity
diameter is variable, between 3.0, 7.6, and 11.7 Å for calix[4]arene, calix[6]arene, and calix[8]arene,
respectively, and between 5.7, 7.8, and 9.5 Å for α, β, and γ-cyclodextrins. Furthermore, calixarenes are
Biomolecules 2019, 9, x FOR PEER REVIEW 10 of 17
highly flexible molecules, while the cyclodextrins are quite rigid molecules. All of the above ensures
Biomolecules 2019, 9, x FOR
good versatility PEER
of the REVIEWof the two macrocycles with the anthelmintic drug.
binding 10 of 17
NO2
OH O
NO2
OH O
NH
NH Cl
Cl
Cl
Cl
Figure 10. Niclosamide structure, an anthelmintic drug used to treat parasitic infections.
Figure 10. Niclosamide structure, an anthelmintic drug used to treat parasitic infections.
Figure 10. Niclosamide structure, an anthelmintic drug used to treat parasitic infections.
The authors indicate that there is an increase of solubility of niclosamide with an increase in
The authors
equal molar indicateofthat
concentrations there isand
calixarene an cyclodextrin,
increase of solubility
but whenof theniclosamide
concentrations with an increase
reach 0.005 M,in
The molar
equal authors indicate thatofthere
concentrations is an increase
calixarene and of solubility
cyclodextrin, but of niclosamide
when the with an reach
concentrations increase 0.005in M,
the solubility decreases, indicating the precipitation of an insoluble complex at higher concentrations.
equal molar
theprocess concentrations
solubility of calixarene
decreases, indicating and cyclodextrin,
the precipitation but
of an when
insoluble the concentrations
complex reach 0.005 M,
The of complexation was explained by hydrogen bonding between theathydroxylic
higher concentrations.
groups of
theThe
solubility decreases, indicating the precipitation of an insoluble complex at higher concentrations.
calixarene and the oxygen atom and the nitrogen atom of niclosamide. Hydrophobic interactionsgroups
process of complexation was explained by hydrogen bonding between the hydroxylic also
Theofprocess of complexation
calixarene andbetween
the oxygen was explained bynitrogen
hydrogen bonding between the hydroxylic groups of
can be suggested the atom and the
hydrophobic cavity of atom of niclosamide.
calixarene Hydrophobic
and the hydrophobic interactions
molecule of
calixarene and
also can be On the oxygen
suggested atom
between and the nitrogen atom of niclosamide. Hydrophobic interactions also
niclosamide. the other hand, thethe hydrophobic cavity of calixarene
solubility of niclosamide and thewith
also increases hydrophobic molecule
the cyclodextrin,
canofbe suggested between
niclosamide. the hydrophobic
On theestablish
other hand, cavity of
the solubility of niclosamide
calixarene and alsothe hydrophobic
increases with molecule of
because these molecules hydrogen bonding with the hydroxyl groups on the the cyclodextrin,
exterior of the
niclosamide.
because theseOn the other hand, the solubility of niclosamide also increases with the cyclodextrin,
cyclodextrin. In molecules
general terms, establish hydrogen
the great bonding
increase with theofhydroxyl
of solubility niclosamide groups on the exterior
is attributed to theof
because these molecules establish hydrogen bonding with the hydroxyl groups on the exterior of the
the cyclodextrin.
additivity of solubilityIn general terms, the great increase
to 4-sulphonato-calix[6]arene andof2-hydroxypropyl-β-cyclodextrin
solubility of niclosamide is attributed [69]. to the
cyclodextrin. In general terms, the great increase of solubility of niclosamide is attributed to the
additivity of solubility to 4-sulphonato-calix[6]arene and 2-hydroxypropyl-β-cyclodextrin [69].
additivity of solubility to 4-sulphonato-calix[6]arene and 2-hydroxypropyl-β-cyclodextrin [69].
2.4.5. Drug Delivery Systems Based on Calixarenes
2.4.5. Drug Delivery Systems Based on Calixarenes
2.4.5. Calixarenes
Drug Deliverycan Systems
also be Based on Calixarenes
useful as nanocarriers in the form of inclusion complexes, micelles,
Calixarenes can also be useful as nanocarriers in the form of inclusion complexes, micelles,
hydrogels, vesicles, liposomes, and nanoparticles. For example, calixarenes have attracted attention
Calixarenes
hydrogels, can also
vesicles, be useful
liposomes, andasnanoparticles.
nanocarriers in Forthe form ofcalixarenes
example, inclusion have complexes,
attractedmicelles,
attention
in medicine for the treatment of cancer, because they can respond to multiple stimuli, are stable, can
hydrogels,
in medicinevesicles, liposomes,
for the treatment and ofnanoparticles.
cancer, because Forthey
example, calixarenes
can respond have attracted
to multiple stimuli,attention
are stable,
avoid nonspecific cell uptake, and are able to reach the target on tumor sites in order to effect the
in medicine
can avoidfor the treatment
nonspecific of cancer,
cell uptake, andbecause
are ablethey can respond
to reach the target toon multiple
tumor stimuli, are stable,
sites in order canthe
to effect
treatment. Calixarenes are ideal for applications in delivery systems, because they have shown good
avoid
treatment. Calixarenes are ideal for applications in delivery systems, because they have shownthe
nonspecific cell uptake, and are able to reach the target on tumor sites in order to effect good
biocompatibility and non-cytotoxicity.
treatment. Calixarenes are ideal
biocompatibility and non-cytotoxicity. for applications in delivery systems, because they have shown good
Cisplatin (Figure 11) is an anticancer agent well known for more than 40 years. There are several
biocompatibility
Cisplatin and non-cytotoxicity.
(Figure 11) is an anticancer agent well known for more than 40 years. There are
examples of formulations of oxaliplatin that use liposomes and micelles as a drug delivery vehicle.
Cisplatin
several (Figureof11)
examples is an anticancer
formulations agent wellthat
of oxaliplatin knownuse for more than
liposomes and40micelles
years. There are several
as a drug delivery
Abbott et al. published a study of the complexation of a dinuclear platinum complex with p-
examples of formulations of oxaliplatin that use liposomes and micelles
vehicle. Abbott et al. published a study of the complexation of a dinuclear platinum complex with as a drug delivery vehicle.
sulphonatocalix[4]arene. The host–guest ratio was 1:1, and the binding constant was 6.8 · 104 M−14. −1
Abbott et al. published a study
p-sulphonatocalix[4]arene. of the complexation
The host–guest ratio was 1:1, of and
a dinuclear
the binding platinum
constant complex
was 6.8 with
· 10 M p- .
sulphonatocalix[4]arene. The host–guest ratio was 1:1, and the binding constant was 6.8 · 10 M . 4 −1

H3 N 2+
Cl
H3N Pt 2+
N Cl
Pt NH3
N
NH NH
3
H3 N NH
N
H3N Pt N
Cl N H
Pt NH3N
Cl H
Figure 11. Chemical structureNH 3
of dinuclear platinum complex trans-[[PtCl(NH3 )2 ]2 µ-dpzm]2+ .

Figure 11. Chemical structure of dinuclear platinum complex trans-[[PtCl(NH3)2]2µ-dpzm]2+.

Figure 11. Chemical structure of dinuclear platinum complex trans-[[PtCl(NH3)2]2µ-dpzm]2+.

The host–guest complex displays no cytotoxicity and is formed by interactions of hydrogen
bonding between the NH3 groups of the metal complex with sulphate groups of the calix[4]arene.
The host–guest complex displays no cytotoxicity and is formed by interactions of hydrogen
 Biomolecules 2019, 9, 90 10 of 15

The host–guest complex displays no cytotoxicity and is formed by interactions of hydrogen

bonding between the NH3 groups of the metal complex with sulphate groups of the calix[4]arene.
Biomolecules 2019, 9, x FOR PEER REVIEW 11 of 17
2.4.6. Controlled Release of Doxorubicin by Vesicles Based on Calixarenes
Because
Because calixarenes
calixarenesare able
are to self-assemble,
able their use
to self-assemble, for the
their useconstruction of vesicles of
for the construction is possible.
vesicles is
Wang et al.Wang
possible. report thereport
et al. construction of binary
the construction supramolecular
of binary supramolecular vesicles driven
vesicles drivenby by
host–guest
host–guest
complexation
complexation between
between a water-soluble
a water-soluble calixarene andand
calixarene viologen. The vesicle
viologen. obtained
The vesicle can be
obtained used
can as
be used
a system of controlled release of doxorubicin hydrochlodire (DOX) (Figure 12),
as a system of controlled release of doxorubicin hydrochlodire (DOX) (Figure 12), a fluorescent dye a fluorescent dye that
is used inused
that is the treatment of cancer.
in the treatment Cell experiments
of cancer. show that
Cell experiments showthethat
release
the of DOXof
release byDOX
the vesicle
by thedoes
vesicle
notdoes
affectnot
theaffect
therapeutic effect of the drug against the cancer cell and by contrast
the therapeutic effect of the drug against the cancer cell and by contrast reduces damage to
reduces
normal
damage cells. The vesicles
to normal are vesicles
cells. The stable for are100 h (approximately
stable 4 days) at4 days)
for 100 h (approximately room at temperature. The
room temperature.
formation of vesicles was followed by ultraviolet–visible (UV–Vis). In the absence
The formation of vesicles was followed by ultraviolet–visible (UV–Vis). In the absence of calixarene, of calixarene, the
maximum absorption of the viologen is 260 nm, and the aggregate is not formed,
the maximum absorption of the viologen is 260 nm, and the aggregate is not formed, but in the presence but in the presence
of of
thethe
macrocycle,
macrocycle, aggregation
aggregation takes place
takes through
place through thethe
formation
formation of of
a complex,
a complex, and andthethe
maximum
maximum
absorption changes to 450 nm. The stability of the vesicle is reinforced by the electrostatic
absorption changes to 450 nm. The stability of the vesicle is reinforced by the electrostatic interaction interaction
of the negative
of the negative groups
groups of of
sulfonate
sulfonate groups
groups of of
resorcinarene
resorcinarene and thethe
and positive
positive groups
groups ofofviologen.
viologen.

O OH O
OH
OH

OH
O O OH O O

OH
NH2
Figure 12. Doxorubicin structure.
Figure 12. Doxorubicin structure.

The obtained vesicles can respond to external stimuli such as temperature, host–guest inclusion,
The obtained vesicles can respond to external stimuli such as temperature, host–guest inclusion,
and redox. In the first case, at temperatures between 5 and 70 ◦ C the processes of assembly and
and redox. In the first case, at temperatures between 5 and 70 °C the processes of assembly and
disassemblyare produced, respectively. The increasing temperature produced the gradual disassembly
disassemblyare produced, respectively. The increasing temperature produced the gradual
of the vesicle. In the second case, taking advantage of the fact that the cyclodextrins can form complexes
disassembly of the vesicle. In the second case, taking advantage of the fact that the cyclodextrins can
with viologen, the gradual addition of α-cyclodextrins was carried out, producing the disruption of the
form complexes with viologen, the gradual addition of α-cyclodextrins was carried out, producing
vesicle. In the last case, it is known that viologens can be transformed from neutral molecules into the
the disruption of the vesicle. In the last case, it is known that viologens can be transformed from
corresponding radical cations by chemical or electrochemical means. The chemical reduction is carried
neutral molecules into the corresponding radical cations by chemical or electrochemical means. The
out with hydrazine, where the solution color changes to purple; however, this does not produce the
chemical reduction is carried out with hydrazine, where the solution color changes to purple;
disruption of the vesicle, but the average diameter decreases from 308 to 153 nm. A similar effect is
however, this does not produce the disruption of the vesicle, but the average diameter decreases from
achieved with electrical redox, but when a reduction potential of −1.6 V vs. Ag/AgCl is applied for
308 to 153 nm. A similar effect is achieved with electrical redox, but when a reduction potential of
30 min, disassembly of the supramolecular vesicle occurs [70].
−1.6 V vs. Ag/AgCl is applied for 30 min, disassembly of the supramolecular vesicle occurs [70].
The above discussion shows that the calixarenes are able to recognize a great diversity of chemical
The above discussion shows that the calixarenes are able to recognize a great diversity of
species, depending on the cavity size and the shape and functional groups of the macrocycle. This
chemical species, depending on the cavity size and the shape and functional groups of the
property is of great importance, because it allows improved bioavailability, solubility, or even activity
macrocycle. This property is of great importance, because it allows improved bioavailability,
of molecules of biological interest. In addition, the calixarenes are able to self-assemble to form micelles,
solubility, or even activity of molecules of biological interest. In addition, the calixarenes are able to
vesicles, liposomes, and nanoparticles that allow the encapsulation, transport, and controlled release
self-assemble to form micelles, vesicles, liposomes, and nanoparticles that allow the encapsulation,
of drugs. Table 1 shows an overview of the aforementioned applications, where the complexation
transport, and controlled release of drugs. Table 1 shows an overview of the aforementioned
with calixarene improved the properties of the biomolecule. Also included are methods, conditions,
applications, where the complexation with calixarene improved the properties of the biomolecule.
and bibliographic references.
Also included are methods, conditions, and bibliographic references.
Biomolecules 2019, 9, 90 11 of 15

Table 1. Calix[n]arenes used for the complexation molecules of biological interest.

Molecule of
Calixarenes Used Method Conditions Ref.
Biological Interest

• The Langmuir film was formed by cellulose acetate and

4-tert-butylcalix[6]arene Electrochemical sensing using 4-tert-butylcalix[6]arene and was transferred to a gold electrode
Dopamine • The detection range was 5–100 and 100–7500 nm and the limit of [62]
Langmuir–Blodgett films
detection was 2.54 nM

4-sulphonic calix[4]arenes,
The concentration of testosterone was determined by
Testosterone 4-sulphonic calix[6]arenes, The solubility of samples were measured at 25 ◦ C at pH 5.8, 7.3, and 10 [63]
HPLC method
4-sulphonic calix[8]arenes.
α-Melanocyte C-tetra(octyl)calixresorcin The surface was characterized by XPS and The peptide was attached to the calixarene with PEG-350 and then coated
[65]
hormone [4]arene MALDI-ToF MS onto glass

• The bindings studied were done at room temperature

5,11,17,23-tetra-tert-butyl-25,26,27,28- • The protein content was analysed via UV–Vis at 476 nm
Green fluorescent The fiber modified was characterized by FTIR, TGA
tetramethoxycarbonylmethoxy- • The synthesis of calix[4]arene tetra ester derivative was carried out on [67]
protein analysis, UV–Vis, fluorescence microscopy and SEM
calix[4]arene diphenyl ether with formaldehyde and a basic medium

The complexation between calixarene-cyclodextrin

Niclosamide Solubility studies were done in a pH 7.0 buffer at 30 ◦ C and ionic strength
4-sulphonatocalix[6]arene and niclosamide was followed by thermal analysis. [67]
(anthelmintic drug) of 0.5 mol/L
The niclosamide content was determined by HPLC

The complex was examined using 1 H nuclear • The complex was formed by an equimolar mixture of dinuclear
Dinuclear platinum complex with p-sulphonatocalix[4]arene
p-sulphonatocalix[4]arene magnetic resonance and electrospray ionization mass [71]
platinum complex • Its water solubility is around 4.5 mM
spectrometry, among others

The nanosupramolecular binary vesicles was studied
by UV–Vis, fluorescence spectroscopy, dynamic laser
Doxorubicin p-sulphonatocalix[4]arene
scattering, transmission electron microscopy, scanning
electron microscopy, and surface tension
• The binary vesicles are formed in the presence of
asymmetric viologen
[70]
• The maximum absorption of the vesicle was at 450 nm

FTIR: Fourier-transform infrared; HPLC: High-performance liquid chromatography; MALDI-ToF MS: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry;
SEM: Scanning electron microscopy; TGA: Thermogravimetric analisys; UV–Vis: Ultraviolet—visible; XSP: X-ray photoelectron spectroscopy.
Biomolecules 2019, 9, 90 12 of 15

3. Conclusions
A wide range of substances of biological interest can be recognized by calix[n]arenes.
The recognition process depends on the size, shape, polarity, type of functional groups present,
aggregation state of the macrocycle, and the formation of non-covalent interactions with the guest.
The design of calix[n]arenes allows for obtaining amphiphilic molecules able to form higher-order
structures such as micelles, vesicles, liposomes, and nanoparticles, which have been shown to be
suitable for controlled release drug delivery systems. In addition, calix[4]aneres are useful for
improving the solubility, biocompatibility, stability, and bioavailability of biomolecules and molecules
with biological activity, and due to the fact that they are not cytotoxic, they can be used in the
manufacture of biomaterials. All the above shows the great potential of these macrocycles in
pharmacology, biomaterials engineering, and medicine, among other fields.

Author Contributions: E.S.E. and M.M.V., designed the research; E.S.E., carried out the bibliographic revision;
M.M.V., performed the collected of the information; E.S.E. and M.M.V., analyzed and organized the results;
E.S.E. and M.M.V., corrected and edited the manuscript; E.S.E. and M.M.V., contributed to the preparation of
the manuscript.
Funding: This research received no external funding.
Acknowledgments: The authors acknowledge to the programs of chemistry of the Universidad Nacional de
Colombia and criminal research of the Universidad Manuela Beltran for their support of this research.
Conflicts of Interest: The authors declare no conflict of interest.

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