U.S. patent application number 17/839851 was filed with the patent office on 2022-09-29 for drug delivery composition comprising a self-assembled gelator.
The applicant listed for this patent is The Brigham and Women`s Hospital, Inc., The City University of New York, represented by The Research Foundation of the City University of NY. Invention is credited to Greg Cruikshank, George John, Jeffrey M. Karp, Praveen Kumar Vemula.
Application Number | 20220305042 17/839851 |
Document ID | / |
Family ID | 1000006402806 |
Filed Date | 2022-09-29 |
United States Patent
Application |
20220305042 |
Kind Code |
A1 |
Karp; Jeffrey M. ; et
al. |
September 29, 2022 |
Drug Delivery Composition Comprising a Self-Assembled Gelator
Abstract
This invention discloses drug-delivery compositions, methods of
making prodrugs, and methods of drug delivery using a
self-assembled gelator. The backbone of the gelator can contain a
drug or prodrug, such as acetaminophen or salicin. Additional drugs
or agents can be encapsulated in the gelator. Enzymatic or
hydrolytic cleavage can be used to release the drugs.
Inventors: |
Karp; Jeffrey M.;
(Brookline, MA) ; Vemula; Praveen Kumar;
(Cambridge, MA) ; John; George; (Edison, NJ)
; Cruikshank; Greg; (Sunnyside, NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
The Brigham and Women`s Hospital, Inc.
The City University of New York, represented by The Research
Foundation of the City University of NY |
Boston
New York |
MA
NY |
US
US |
|
|
Family ID: |
1000006402806 |
Appl. No.: |
17/839851 |
Filed: |
June 14, 2022 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15213055 |
Jul 18, 2016 |
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17839851 |
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13119304 |
Jun 6, 2011 |
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PCT/US2009/057349 |
Sep 17, 2009 |
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15213055 |
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61097565 |
Sep 17, 2008 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/405 20130101;
A61K 9/06 20130101; A61K 31/43 20130101; A61K 45/06 20130101; A61K
47/542 20170801; A61K 31/12 20130101; A61K 31/7034 20130101; A61K
31/22 20130101; A61K 47/55 20170801; A61K 47/54 20170801; A61K
31/555 20130101; A61K 47/555 20170801; A61K 31/167 20130101 |
International
Class: |
A61K 31/7034 20060101
A61K031/7034; A61K 9/06 20060101 A61K009/06; A61K 31/167 20060101
A61K031/167; A61K 45/06 20060101 A61K045/06; A61K 31/12 20060101
A61K031/12; A61K 31/555 20060101 A61K031/555; A61K 31/22 20060101
A61K031/22; A61K 31/405 20060101 A61K031/405; A61K 31/43 20060101
A61K031/43; A61K 47/55 20060101 A61K047/55; A61K 47/54 20060101
A61K047/54 |
Claims
1. A drug-delivery composition, comprising a drug-derived gelator
capable of self assembly, wherein the backbone of the gelator
contains a drug a derivative thereof and which drug can be released
upon a stimuli.
2. The composition of claim 1, wherein the stimuli is
enzyme-mediated degradation or hydrolytic cleavage.
3. The composition of claim 1, wherein the stimuli is of biological
origin.
4. The composition of claim 1, wherein the biological origin is
inflammation.
5. The composition of claim 1, wherein the composition is a
hydrogel.
6. The composition of claim 1, wherein the composition is
self-assembling.
7. The composition of claim 1, wherein the drug-derived gelator is
a self-assembled gelator.
8. The composition of claim 7, wherein the gelator is a hydrogel or
an organogelator.
9. The composition of claim 1, wherein the backbone of the
drug-derived gelator contains the drug or a derivative thereof, a
hydrophobic moiety, and a degradable link between the drug and the
hydrophobic moiety.
10. The composition of claim 1, wherein the drug is a hydrophobic
drug or hydrophilic drug.
11. The composition of claim 1, wherein backbone of the
drug-derived gelator comprises at a hydrophobic drug and a
hydrophilic moiety.
12. The composition of claim 1, wherein the drug-derived gelator
comprises a hydrophilic drug and a hydrophobic moiety.
13. The composition of claim 9, wherein the drug-derived gelator is
a drug derived, self-assembled gelator.
14. The composition of claim 9, wherein the hydrophobic moiety is a
polymethylene chain or an aromatic group.
15. The composition of claim 9, wherein the degradable link is an
ester, amide, anhydride, or carbamate linkage that is cleavable
upon contact with an enzyme and/or through hydrolysis.
16. The composition of claim 1, wherein the stimuli is temperature,
pH, ultrasound, metal ions, light, electrical stimuli,
electromagnetic stimuli, or combinations thereof.
17. The composition of claim 1, wherein the gelator is the
esterification reaction product of the drug and a C.sub.1-22
carboxylic acid.
18. The composition of claim 17, wherein the carboxylic acid
contains saturated or unsaturated hydrocarbon chains.
19. The composition of claim 1, wherein the drug contains a phenol
ring and a terminal hydroxyl group, and wherein the drug is capable
of undergoing self assembly upon esterification with a fatty acid
or dicarboxylic acid.
20. The composition of claim 1, wherein the drug contains one or
more functional groups that promote self-assembly through pi-pi
stacking, hydrogen-bonding and/or van der Waals interactions.
21.-166. (canceled)
Description
RELATED APPLICATIONS
[0001] This application is a continuation of prior U.S. patent
application Ser. No. 15/213,055, filed Jul. 18, 2016, which in turn
is a continuation of prior U.S. patent application Ser. No.
13/119,304, filed Jun. 6, 2011, which in turn is a National Stage
of International Patent Application Serial No. PCT/US2009/057349,
filed Sep. 17, 2009, which in turn claims the benefit of priority
to U.S. Provisional Patent Application Ser. No. 61/097,565, filed
Sep. 17, 2008, the contents of which are incorporated by reference
herein in their entirety.
FIELD OF THE INVENTION
[0002] This invention relates to low-molecular-weight amphiphilic
gelators which self-assemble into various nano- and
micro-structures in a wide range of organic and aqueous solvents.
The gelators have the capability to deliver drugs.
BACKGROUND
[0003] A hydro- or organo gel consists of self-assembled
macromolecular networks with a liquid filling the interstitial
space of the network. The network holds the liquid in place through
its interaction forces and so gives the gel solidity and coherence,
but the gel is also wet and soft and capable of undergoing some
extent of deformation. The gel state is neither solid nor liquid
but has some features of both. Self-assembly has been used to
develop molecularly defined and functional materials, including
hydrogels.
[0004] Self-assembled hydrogels can be formulated in a variety of
physical forms, including microparticles, nanoparticles, coatings
and films. As a result, hydrogels are commonly used in clinical
practice and experimental medicine for a wide range of
applications, including tissue engineering and regenerative
medicine, diagnostics, cellular immobilization, separation of
biomolecules or cells and barrier materials to regulate biological
adhesions. Hydrogels are appealing for biological applications
because of their high water content and biocompatibility. In the
last few decades hydrogels have gained considerable attention, and
significant progress has been made in designing, synthesizing and
using these materials for several biomedical applications including
drug delivery. Recent developments include the design and synthesis
of novel hydrogels and their use in tissue engineering, drug
delivery and bionanotechnology.
[0005] Design and synthesis of low-molecular-weight hydrogelators
has received considerable attention in soft materials research in
terms of its potential use in cosmetics, toiletries and
pharmaceutical formulations. Literature study reveals that there
are only limited reports on easily achievable and efficient
low-molecular-weight gelators that are able to gel water or even
water mixtures with other solvents. These hydrogelators are often
achieved by multi-step chemical syntheses. See Menger, F. M. &
Caran, K. L. (2000) J. Am. Chem. Soc. 122, 11679-11691;
Sreenivasachary, N. & Lehn, J.-M. (2005) Proc. Natl. Acad. Sci.
USA 102, 5938-5943; Makarevi , J., Joki , M., Perci , B., Tomis i ,
V., Kroji -Prodi , B. & Z ini , M. (2001) Chem. Eur. J. 7,
3328-3341; Oda, R., Huc, I. & Candau, S. J. (1998) Angew. Chem.
Int. Ed. 37, 2689-2691; Estroff, L. A. & Hamilton, A. D. (2000)
Angew. Chem. Int. Ed. 39, 3447-3450; Kobayashi, H., Friggeri, A.,
Koumoto, K., Amaike, M., Shinkai, S. & Reinhoudt, D. N. (2002)
Org. Lett. 4, 1423-1426; Luboradzki, R., Gronwald, O., Ikeda, M.,
Shinkai, S. & Reinhoudt, D. N. (2000) Tetrahedron 56,
9595-9599; Jung, J. H., John, G., Masuda, M., Yoshida, K., Shinkai,
S. & Shimizu, T. (2001) Langmuir 17, 7229-7232; and Wang, G.
& Hamilton, A. D. (2003) Chem. Commun. 310-311.
[0006] Low molecular weight gelators are attractive alternatives to
their polymeric counterparts as such molecules can be
systematically altered and explain various factors which are
responsible for the gelation at microscopic level. Because of their
importance as scaffolds for various biomaterials design, it is
useful to develop low-molecular-weight gelators that ensure strong
gelation of water under ambient conditions.
[0007] Synthesizing hydrogelators in high quantities is often
limited with multiple-step synthesis and poor yields, which make
industrial scale production difficult. To develop hydrogelators in
large scales, one needs to minimize the number of steps in the
synthesis and reduce the side products. Many reported hydrogelators
require multiple steps and an elaborated purification process, and
many others require costly starting materials and expensive
reagents.
[0008] Thus, there exists a need in the art for an improved gelator
that is easily prepared and can be effectively used for drug
delivery. This invention answers that need.
[0009] An additional problem in drug delivery is striking a balance
between toxicity and therapeutic effect. Hydrogels have been widely
applied as carriers in controlled drug delivery systems. See
Hydrogels in Medicine and Pharmacy (Ed: Peppas, N.) CRC, Boca
Raton, Fla. 1987; Peppas, N. A.; Bures, P.; Leobandung, W.;
Ichikawa, H. Eur. J. Pharm. Biopharm. 2000, 50, 27-46; Peppas, N.
Curr. Opin. Colloid Interface Sci. 1997, 2, 531-537; Gupta, P.;
Vermani, K.; Garg, S. Drug Discovery Today 2002, 7, 569-579;
Miyata, T.; Uragami, T.; Nakamae, K. Adv. Drug Deliv. Rev. 2002,
54, 79-98; Qiu, Y.; Park, K. Adv. Drug Deliv. Rev. 2001, 53,
321-339; and Kamath, K.; Park, K. Adv. Drug Deliv. Rev. 1993, 11,
59-84. However, many of the systems are derived from polymeric
hydrogels as opposed to hydrogels formed through self-assembly of
low-molecular-weight gelators (LMWGs). This could be due to the
relatively complex synthesis of LMWGs and concerns about possible
toxicity of in situ generated fragments from the degradation of
LMWGs. Additionally, self-assembled hydrogel-based drug delivery
has been hindered by the unknown fate of the host gelator after the
gel degradation.
[0010] Potential problems can arise relating to the quantity of
drug that can be encapsulated within hydrogels, and respective
encapsulation efficiency may limit the potential dose or release
kinetics that can be achieved. Additionally, polymer-based drug
delivery devices that utilize entrapped drug have been known to
exhibit an undesired burst release. See Huang, X.; Brazel, C. S. J.
Controlled Release 2001, 73, 121-136.
[0011] In an attempt to address these problems, small molecule
drugs have been incorporated into the backbone of polymers. See
Bryers, J. D.; Jarvis, R. A.; Lebo, J.; Prudencio, A.; Kyriakides,
T. R.; Uhrich, K. Biomaterials 2006, 27, 5039-5048; Erdmann, L.;
Macedo, B.; Uhrich, K. E. Biomaterials 2000, 21, 2507-2512; and
Harten, R. D.; Svach, D. J.; Schmeltzer, R.; Uhrich, K. E. J.
Biomed. Mater. Res-A 2005, 72A, 354-362. However, polymer
degradation can lead to polymer fragments with heterogeneous chain
lengths that may generate potential toxicity, unwanted side
effects, or uncontrollable drug bioavailability.
[0012] While progress in this field has been made (see Vemula, P.
K.; Li, J.; John, G. J. Am. Chem. Soc. 2006, 125, 8932-8938), there
is an ongoing need to develop degradable prodrug-based
self-assembled gelators that can overcome these problems. This
invention also answers that need.
SUMMARY OF THE INVENTION
[0013] One embodiment of this invention relates to drug-delivery
compositions. The drug may be delivered in various different ways.
The drug-delivery composition may be a drug-derived gelator capable
of self assembly, where the drug to be delivered is in the backbone
of the gelator and released upon degradation of the gelator. One or
more agents can be encapsulated in the self-assembled gels.
Degradation of the gelator can release both the encapsulated drugs
and the drug in the backbone of the gelator. In this aspect, the
drug-delivery composition is a multi-drug delivery composition. The
composition may also be a self-assembled gelator that is not drug
derived but still contains one or more drugs encapsulated in the
gelator that can be released upon degradation of the gelator.
Prodrug-derived compounds, such as prodrug-derived self-assembled
gelators may also be used.
[0014] Another embodiment of this invention relates to a
single-step method of making a low molecular weight, amphiphilic
prodrug or gelator. Prodrugs may be prepared by esterifying a
compound having groups capable of promoting self assembly, such as
a phenol ring and/or a terminal hydroxyl group, with a dicarboxylic
acid to form a prodrug having an ester functionality. Other
prodrugs may be prepared by esterifying a compound having a
saccharide group, phenol ring, and a primary hydroxyl group with a
vinyl ester of a carboxylic acid. Gelators may be prepared by
reacting a tris-hydroxyl methyl amine, such as tris, with a fatty
acid or methyl ester to form a gelator having an amide
functionality.
[0015] Another embodiment of this invention relates to methods of
drug delivery. A drug may be delivered to a patient by
administering the patient a drug-derived or prodrug-derived gelator
capable of self assembly, the gelator containing the drug or
prodrug in its backbone, a hydrophobic or hydrophilic moiety, and a
degradable link between the drug and the moiety. When contacted
with an enzyme under physiological conditions, the degradable link
cleaves, releasing the drug in the patient. Hydrolytic cleavage
under basic conditions can also be used. If the gelator contains an
agent, such as another drug, that drug can also be released upon
cleavage of the link. A drug may also be administered by
encapsulating the drug in a self-assembled gelator, where the
gelator may contain an amide functionality and hydrophobic moiety.
When contacted with an enzyme under physiological conditions, the
amide bond is cleaved, releasing the encapsulated drug.
BRIEF DESCRIPTION OF THE DRAWINGS
[0016] FIG. 1 is a schematic representation of preparing degradable
prodrug-based hydrogelators, encapsulation of hydrophobic drug in
the gel, and subsequent enzyme triggered single (path-1) and
multiple (path-2) drug delivery.
[0017] FIG. 2 is a schematic representation of synthesis of
amphiphiles and bolaamphiphiles from acetaminophen.
[0018] FIGS. 3A-3D depict SEM images of hydrogels made from
Apn-7-COOH (3A) and Apn-8-Apn (3B). Unstained TEM images of
hydrogels made from Apn-7-COOH (3C) and Apn-8-Apn (3D). FIG. 3E
shows the real image of the hydrogel.
[0019] FIG. 4A depicts optimized geometries of hydrogelators. FIGS.
4B-4D depict proposed mode of self-assembly into fiber based
hydrogels made of Apn-7-COOH (4B) and Apn-8-Apn (4C and 4D).
[0020] FIGS. 5A-5D depict SEM images of curcumin encapsulated
hydrogels made from Apn-7-COOH (5A) and Apn-8-Apn (5B). Unstained
TEM images of curcumin encapsulated hydrogels made from Apn-7-COOH
(5C) and Apn-8-Apn (5D). FIG. 5E shows the real image of the
hydrogel.
[0021] FIG. 6A depicts a schematic representation of
enzyme-mediated single and two drugs delivery from Apn-based
prodrug hydrogels. FIG. 6B shows absorbance spectra for methanolic
solution of hydrogelators and curcumin. FIG. 6C shows absorption
spectra of curcumin released from degradation of Apn-7-COOH
hydrogel at different time points, i) 4, ii) 8, iii) 12, iv) 16, v)
20 and vi) 24 hrs (after 24 hrs absorbance did not increase). FIG.
6D is chemical structure of curcumin, and FIG. 6E is absorbance
maxima of curcumin vs time.
[0022] FIGS. 7A-7D depict bar graph showing the effect of Apn-based
prodrugs on MSCs cellular properties such as viability (7A and 7B),
adhesion (7C), and proliferation (7D). Cell viability was examined
after 48 hours (7A and 7B), adhesion was measured after 12 hours
(7C). Concentration of Apn and other amphiphiles used in FIGS. 7A,
7C, and 7D are 250 .mu.M.
[0023] FIGS. 8A-8E depict results of adipogenesis after 23 days of
induction as observed by Oil Red O (ORO) staining. Controls cells
treated with phosphate buffer saline (PBS) (8A) and 250 .mu.M Apn
(8B) showed positive staining. Cells treated with 250 .mu.M of
Apn-7-COOH (8C) and Apn-8-Apn (8D) were also equally stained for
ORO. FIG. 8E shows the magnified region of fat droplets. In all
images scale bar=100 .mu.m.
[0024] FIGS. 9A-9E depict results of osteogenesis after 23 days as
observed by alkaline phosphatase (ALP) staining. Controls cells
treated with phosphate buffer saline (PBS) (9A) and 250 .mu.M Apn
(9B) were showed positive staining. Cells treated with 250 .mu.M of
Apn-7-COOH (9C) and Apn-8-Apn (9D) were also equally stained for
ALP. FIG. 9E shows the magnified region of differentiated cells,
and black spots are mineralized foci localized to ALP stained
cells. In all images scale bar=100 .mu.m.
[0025] FIGS. 10A-10C illustrate the self-assembly of
acetaminophen-derived gelators (depicted in FIG. 10A) encapsulating
a second drug (depicted in FIG. 10B).
[0026] FIGS. 11A-11B depict SEM images of (11A) organo and (11B)
hydrogels formed by self-assembly of polyol-based amphiphiles.
[0027] FIG. 12 is a schematic representation of self-assembly of
amphiphiles to form gels and encapsulation of drugs in their
assembled structures.
[0028] FIGS. 13A-13B depict bar graphs showing normalized
fluorescence intensity of encapsulated dye within the fibers (13A)
in the absence and (13B) in the presence in the presence of
synovial fluid that was collected from inflamed human arthritis
joints.
[0029] FIGS. 14A-14F depict depicts optical microscope images of
arthritis bearing mice joints (row-1 knees (14A-14C), row-2 ankles
(14D-14F)) with DiD encapsulated self-assembled nanofibers.
[0030] FIG. 15 is a schematic representation of Apn showing atom
numbering used to report selected geometric parameters.
DETAILED DESCRIPTION
Drug-Delivery Compositions
[0031] This invention relates to a drug-delivery composition
comprising a drug-derived gelator capable of self assembly. After
the drug-derived gelator has undergone self assembly, the gelator
becomes a self-assembled gelator. The drug to be delivered is part
of the backbone of the gelator and can be released upon
enzyme-mediated degradation or through hydrolytic cleavage of the
gelator. Alternatively, the drug used in the compound may be a
prodrug.
[0032] Thus, an embodiment of the invention relates to
prodrug-delivery composition comprising a prodrug-derived gelator
capable of self assembly where the prodrug to be delivered is part
of the backbone of the gelator and can be released under the
appropriate conditions, e.g., enzyme-mediated degradation or
through hydrolytic cleavage of the gelator. Like the drug-derived
gelator, the prodrug-derived gelator becomes a self-assembled
gelator upon self assembly. Any drug or prodrug known to those of
skill in the art may be used with this invention, provided that the
drug or prodrug, and/or gelator is biocompatible with the host that
the drug-delivery composition is being administered to.
[0033] As used herein, the term "drug" means a molecule, group of
molecules, complex or substance administered to an organism for
diagnostic, therapeutic, medical or veterinary purposes. Drugs
include externally and internally administered topical, localized
and systemic human and animal pharmaceuticals, treatments,
remedies, nutraceuticals, cosmeceuticals, biologicals, devices,
diagnostics and contraceptives, including preparations useful in
clinical and veterinary screening, prevention, prophylaxis,
healing, wellness, detection, imaging, diagnosis, therapy, surgery,
monitoring, cosmetics, prosthetics and forensics. In certain
embodiments, the drug is hydrophilic. In certain other embodiments,
the drug hydrophobic.
[0034] As used herein the term "prodrug" means a drug, drug
precursor of modified drug that is not fully active or available
until converted in vivo or in situ to its therapeutically active or
available form.
[0035] In addition to containing the drug or prodrug, or a
derivative thereof, the backbone of the drug-derived or
prodrug-derived gelator may also contain a hydrophobic or
hydrophilic moiety, and a degradable link between the drug or
prodrug and the moiety. For example, if the drug or pro-drug is
hydrophilic then the moiety is hydrophobic and if the drug or
pro-drug is hydrophobic then the moiety is hydrophilic. Without
wishing to be bound by theory, the having both a hydrophilic
component and a hydrophobic component allows the gelator to
self-assemble under appropriate conditions. Any degradable link
that is cleavable under the required conditions, i.e. a stimuli,
can be used. Preferably, the degradable link is cleaved at least 10
times or more, preferably at least 100 times faster under the
required conditions or under a first reference condition (which
can, e.g., be selected to mimic or represent intracellular
conditions) than in the blood of a subject, or under a second
reference condition (which can, e.g., be selected to mimic or
represent conditions found in the blood or serum).
[0036] The stimuli can be of biological origin. As used herein, the
term "biological origin" refers to conditions that are present in
biological systems. In certain embodiments, the degradable link is
cleavable under biological conditions, e.g., conditions present in
the blood or serum, or conditions present inside or out side the
cell, tissue or organ. The degradable link can be cleavable only
under conditions present in a disease state of a cell, tissue or
organ, e.g. inflammation. Without wishing to be bound by theory,
the link is cleaved when appropriate conditions are present for the
cleavage, e.g. pH or concentration. Thus allowing for release of
drug at targeted tissue and/or organ. This can allow use of lower
dosage because the drug is only released at the required site.
Another benefit is lowering of toxicity to other organs and
tissues.
[0037] Without wishing to be bound by theory, cleavage of the link
under biological conditions allows self-titering drug delivery. As
used herein, the term "self-titering" refers to release of drug
based on the severity of disease or inflammation. By way of
explanation, when the severity of disease is severe, stimuli for
cleaving the degradable link is high and more drug is released and
when severity of disease is low, stimuli is low and less drug is
released. Thus, for example, in a strong inflammatory response,
there would be more enzymes present and this would lead to higher
release of the drug, and when the inflammation subsides, i.e.
remission, the drug would not be released.
[0038] In certain embodiments of the invention, any degradable link
that is cleavable upon contact with an enzyme and/or through
hydrolysis can be used. Examples of degradable links include ester,
amide, anhydride, and carbamate linkages. Suitable hydrophobic
moieties include phenol rings, hydroxyl groups, polymethylene
chains, amides, carbonyls, and carboxylic acids.
[0039] In certain embodiments, the degradable link can be phosphate
based, e.g. phosphodiester. Phosphate-based linking groups are
cleaved by agents that degrade or hydrolyze the phosphate group. An
example of an agent that cleaves phosphate groups is
phosphatases.
[0040] In some embodiments, the degradable link can be linking
group that is cleaved under acidic conditions. In preferred
embodiments acid cleavable linking groups are cleaved in an acidic
environment with a pH of about 6.5 or lower (e.g., about 6.0, 5.5,
5.0, 4.5, 4.0 or lower), or by agents such as enzymes that can act
as a general acid. In a cell, specific low pH organelles, such as
endosomes and lysosomes can provide a cleaving environment for acid
cleavable linking groups. Examples of acid cleavable linking groups
include but are not limited to hydrazones, esters, and esters of
amino acids. Acid cleavable groups can have the general formula
--C.dbd.NN--, --C(O)O--, or --OC(O)--. Another degradable link that
is also susceptible to low pH is disulfide (--S--S--).
[0041] A degradable link can be susceptible to temperature, for
example cleavable at high temperature, e.g., cleavable in the
temperature range of 37-10, 40-100, 45-100, 50-100, 60-100,
70-100.degree. C. For example, the linkage can be cleaved by an
increase in temperature.
[0042] Other degradable links are redox cleavable and are cleaved
upon reduction or oxidation. One reductively degradable link is a
disulphide linking group (--S--S--).
[0043] In certain embodiments, stimuli can be ultrasound,
temperature, pH, metal ions, light, electrical stimuli,
electromagnetic stimuli, and combinations thereof
[0044] In certain embodiments, the drug-derived or prodrug-derived
gelator comprises a hydrophilic drug and a hydrophobic moiety. In
other embodiments, the drug-derived or prodrug-derived gelator
comprises a hydrophobic drug and a hydrophilic moiety.
[0045] The gelator can either be a hydrogelator or organogelator.
Hydrogels, as known to those of skill in the art, are 3-D networks
of molecules typically covalently or ionically linked together
where water is the major component (usually greater than 80%). Gels
can be formed via self-assembly of gelators or via chemical
crosslinking of gelators. Water-based gelators can be used to form
hydrogels, whereas organogelators are gelators that form gels
(organogels) in organic solvents. In one embodiment, the gelator is
the esterification reaction product of the drug or prodrug and a
C.sub.1-22 carboxylic acid. Preferably, the carboxylic acid is a
dicarboxylic acid. The carboxylic acid may contain saturated or
unsaturated hydrocarbon chains.
[0046] Any drug or prodrug that is capable of undergoing self
assembly may be used as the drug or prodrug in the drug-derived or
prodrug-derived gelator. In one embodiment, the drug or prodrug is
capable of undergoing self assembly upon esterification with a
fatty acid or dicarboxylic acid. The self-assembled gelator may be
a single-component gelator that does not require any other
complementary molecules or base salts to induce self assembly.
[0047] Preferably, the drug or prodrug used in the gelator is
easily functionalizable without compromising the base structure of
the drug. Functionalizable groups include hydroxyls, amines,
carboxylic acids, thiols, anhydrides, and others known to one of
skill in the art. Either the drug or the group that is conjugated
to the drug may have particular moieties that can be used to induce
self-assembly. In general, phenol rings, hydroxyl groups,
polymethylene chains, amides, carbonyls, or carboxylic acids may be
used to aid in the self-assembly process. For instance, the drug
used in the gelator may contain a phenol ring and a terminal
hydroxyl group. The drug may also contain other functional groups
that promote self assembly, in particular self assembly that occurs
upon esterification with a fatty acid or dicarboxylic acid.
[0048] There are different ways to promote self assembly. The
functional groups of the gelator and/or drugs can promote self
assembly by facilitating the ability of the gelator to interact
non-covalently with other gelator molecules, for instance by
facilitating hydrogen bonding, intermolecular interactions through
pi-pi stacking, and/or molecular association through van der Waals
interactions.
[0049] The other significant parameter that promotes self-assembly
is the solubility of the gelator, where the solubility properties
of gelators can play a significant role in determining gelation
efficiency. As known to those of skill in the art, self-assembly is
typically driven by a stimulus that alters the solubility of the
gelator. A change in solubility can be induced by a variety of
techniques known to those of skill in the art, including changing
the temperature of the solvent/gelator mixture. The solubility
properties of many different drugs can thus be modulated by
appropriate functionalization with various moieties to tailor the
solubility and self-assembling properties.
[0050] Various methods and techniques can be used to achieve
dissolution of gelators/prodrug/amphiphile in aqueous solution or
organic solvents, including altering the temperature, sonication,
grinding, co-solvent method, mixed solvent method, and combinations
thereof. For example, a drug that is soluble in water may be
resistant to induce self-assembly in water. A hydrophobic tail, for
instance a polymethylene chain, could be added to the drug. Most
hydrophobic tails will not be soluble in water at room temperature,
yet will solubilize when the water is heated. When the composition
cools, the gelator precipitates in a highly organized
self-assembled fashion. These techniques can be used for
hydrogelator and organogelators alike, particularly in instances
where it is desirable that the gelator initially not be soluble in
a particular solvent.
[0051] Others methods may also be used to modify solubility. For
example, certain prodrugs may dissolve in a solvent like DMSO but
not water. In this instance, the prodrug could first be dissolved
in DMSO, or any other water-miscible solvent, then placed into
water (a non-solvent for the prodrug). Because DMSO is miscible
with the water, the prodrug will become supersaturated and
precipitate into a highly organized self-assembled state. In these
type of scenarios, the alterations to the drug would not be
temperature dependent, as discussed above, but may be used just as
effectively to increase solubility in one solvent and not in
another.
[0052] Using these various techniques and methods to achieve
self-assembly enables self assembly to occur in vitro or in situ.
For instance, injecting the gelator into warm solution and as it
cools (typically to 37.degree. C.) would promote self assembly.
Alternatively, the gelator could be injected with a suitable
water-miscible solvent and as this solvent mixes with water in the
tissue, the gelator would self assemble. Thus, injectable or
implantable methods and techniques can also be used to promote self
assembly of the gelator.
[0053] The drug-based gelators and prodrug-based gelators of this
invention can be based on various drugs or prodrugs known to those
of skill in the art. The drug used in the gelator preferably
contains a phenol group and a terminal hydroxyl group. More
preferably, the drug is acetaminophen or a derivative of
acetaminophen, and the gelator is an acetaminophen-derived gelator.
Most preferably, the acetaminophen-derived gelator is selected from
formula (I) or formula (II):
##STR00001##
wherein n in each formula represents an integer from 1-22.
Preferably n is 1-12 in both formula (I) and formula (II), with
6-12 being the most preferred range of n.
[0054] The prodrug, while similar to the drug, preferably contains
a phenol group, a terminal hydroxyl group, such as hydroxymethyl
group, and a saccharide or a saccharide derivative such as
poly-saccharide. In a preferred embodiment, the prodrug is salicin
or a derivative of salicin and the gelator is a salicin-derived
gelator. Most preferably, the prodrug-derived gelator is of formula
(III):
##STR00002##
where R is a saturated or unsaturated C.sub.1-21 alkyl group.
[0055] The drug-delivery composition may also contain an agent
encapsulated in the self-assembled gels that can be released with
the drug or prodrug upon enzyme-mediated degradation or hydrolytic
cleavage of the gelator. Suitable agents include organic dyes,
vitamins, derivatives of vitamins, peptides, proteins, enzymes,
RNAi, chemotherapeutics, anti-inflammatory drugs, and combinations
thereof. Preferably, the agent is another drug, an enzyme, or both
an enzyme and another drug.
[0056] When one or more drugs are encapsulated in the gelator, the
drug-delivery composition is capable of delivery two or more drugs
through the composition. Thus, an embodiment of the invention
relates to a multi-drug delivery composition that contains a
self-assembled gelator containing a first drug or prodrug and a
second drug. The first drug or prodrug, as described above, can be
part of the backbone of the gelator and the second drug can be
encapsulated in the gel of the gelator. Upon exposure to a suitable
enzyme or upon hydrolytic cleavage of the gelator, the first drug
and the second drug can be simultaneously or nearly simultaneously
released. It is also possible for the composition to further
contain one or more additional drugs in the backbone of the gelator
and/or one or more additional drugs encapsulated in the gel of the
gelator.
[0057] In a preferred embodiment, the first drug is an analgesic or
antipyretic drug and the second drug is an anti-inflammatory agent,
or vice versa. The two drugs may be complementary to one another.
For instance, both drugs may be designed to treat the same ailment
or both drugs may be routinely prescribed by a physician to treat
different aspects of the same ailment. Both drugs may be delivered
in a sustained release. When the drug is a hydrophobic drug, it can
be encapsulated in the hydrophobic pockets of the self-assembled
gelator, making for particularly effective encapsulation.
[0058] Drugs may also be delivered through a self-assembled gelator
when the composition is not derived from a drug or prodrug. This
embodiment of the invention relates to a drug-delivery composition
that contains a drug and a self-assembled gelator having an ester,
amide, or carbamate functionality and a hydrophobic moiety in the
backbone of the gelator. The drug is encapsulated in the gel and
can be released upon enzyme-mediated degradation of the gelator or
hydrolytic cleavage of the gelator.
[0059] In a preferred embodiment, the gelator is a tris-based
hydrogelator having an amide functionality. Exemplary gelators are
those of formula (IV):
##STR00003##
wherein R is a saturated or unsaturated C.sub.1-22 alkyl group, and
R.sup.1, R.sup.2, and R.sup.3 each independently represent a
hydroxyalkyl group, such as a hydroxy C.sub.1-6 alkyl. Preferably,
R is a saturated or unsaturated C.sub.9-21 alkyl group (more
preferably C.sub.12-16), and R.sup.1, R.sup.2, and R.sup.3 each
represent a hydroxymethyl group. More preferably, the gelator is
the reaction product of 2-amino-2-hydroxymethylpropane-1,3-diol
(tris) and a fatty acid or methyl ester of a fatty acid. Tris, as
known to those of skill in the art, is widely used as a component
of buffer solutions. In particular, it is useful to act as a buffer
to maintain tissue microenvironment pH, typically at a pH between
7-9.
[0060] Certain enzymes have been found to be particularly useful
for tris-based hydrogelators. For instance, enzymes in the amidase
or metalloprotease families of enzymes have been found to be useful
at cleaving amide bonds.
[0061] Another embodiment of this invention relates to a
single-component drug-based gelator composition, where the gelator
contains functional groups that exist within the drug and/or within
a modified moiety that is conjugated to the drug. The functional
groups can interact via pi-pi stacking, hydrophobic interactions,
and/or hydrogen bonding. As discussed above, the drug can be
released enzymatically and/or hydrolytically.
[0062] Compositions of the invention can have a variety of
different shapes and structures. For example, the compositions of
the invention can be formed into capsules, tablets, films,
microspheres or the like. In certain embodiments, the compositions
of the invention are particles e.g. nanoparticles or
microparticles.
[0063] In certain embodiments, the composition is a particle having
one dimension in the range of 10,000-100,000 nm. In other
embodiments, the composition is a particle having one dimension in
the range of 1000-10,000 nm. In certain embodiments, the particles
have one dimension in the range of 1-3000 nm.
[0064] In some embodiments, the particles have one dimension in the
range of 1-1000 nm. Preferably the particles have one dimension in
the range of 20-750 nm. More preferably the particles have one
dimension in the range of 20-500 nm. And most preferably the
particles have one dimension in the range of 50-500 nm.
[0065] In certain embodiments, the nano- or micro-particles are a
hydrogel comprising prodrug-derived gelator. In certain further
embodiments of this and other aspects of the invention, components
of the hydrogel are non covalently bound with each other.
[0066] As discussed above, drugs can also be encapsulated in the
compositions of the invention. In certain embodiments, the drug is
indomethacin or ethambutol. In some embodiments, indomethacin or
ethambutol is connected to a peptide, fatty acid or sugar
moiety.
[0067] In some embodiment, the indomethacin is of formula (V):
##STR00004##
wherein R is peptide comprising 1-40 amino acids or R is saturated
or unsaturated C.sub.1-C.sub.21 alkyl group. In some embodiments, R
is C.sub.1-C.sub.6 alkyl. In some other embodiments, R is
C.sub.6-C.sub.12 alkyl.
[0068] In some embodiments, the ethambutol is of formula (VI):
##STR00005##
wherein R is a peptide comprising 1-40 amino acids or R is
saturated or unsaturated C.sub.1-21 alkyl group. In some
embodiments, R is C.sub.1-C.sub.6 alkyl. In some other embodiments,
R is C.sub.6-C.sub.12 alkyl.
[0069] In certain embodiments, the drug promotes specific cellular
function.
[0070] In certain embodiments, the encapsulated drug is a small
molecule that can promote differentiation, de-differentiation,
trans-differentiation, or reprogramming of cells.
[0071] The compositions of the invention can comprise two or more
different gelators. For example, the two or more different gelators
can differ from each other by drug or prodrug, hydrophobic or
hydrophilic moieties present in the backbone.
[0072] In certain embodiments, the composition comprises two
different gelators.
[0073] In certain embodiments, hydrogel can by lyophilized. Without
wishing to be bound by theory, lyophilized gels can be formulated
for aerosol based drug delivery.
[0074] It is to be understood that upon release from the
composition, drug is released in its therapeutically active form.
This applies to both the drugs or prodrugs that are part of the
gelator backbones and the drugs that are encapsulated within the
gel and are not part of the backbone.
[0075] As one of skill in the art can appreciate, the precise
amount of drug that is available for release is pre-defined by the
composition of the drug or prodrug used in the gelator. Without
wishing to be bound by theory, the amount available for release can
be increased or decreased by changing the composition of the
gelator.
[0076] The compositions of the invention are particularly useful,
e.g., for improving safety, targeting efficiency, compliance and
efficacy for indications benefiting from single-dose, prolonged
action or tissue-specific formulations. Exemplary indications
include, but are not limited to, allergy (e.g. contact dermatitis),
arthritis, asthma, cancer, cardiovascular disease, diabetic ulcers,
eczema, infections, inflammation, muscuscitis, periodontal disease,
psoriasis, respiratory pathway diseases (e.g., tuberculosis),
vascular occlusion, pain.
[0077] The compositions of the invention are also useful for oral
drug delivery, inner ear drug delivery, transdermal drug delivery
(e.g., permeabilization), or an anti-bacterial. Furthermore, the
compositions can be used to deliver drugs for treatment of pain and
graft versus host diseases.
Method of Making Prodrug and Gelator
[0078] An embodiment of this invention relates to methods of making
prodrugs and gelators. One embodiment relates to a single-step
method of making a low molecular weight, amphiphilic prodrug,
comprising the step of esterifying a compound having a phenol ring
and terminal hydroxyl group with a dicarboxylic acid, for instance
a C.sub.1-22 dicarboxylic acid, to form an amphiphilic prodrug
containing an ester functionality. The esterification reaction can
take place without the use of a catalyst.
[0079] The prodrug may then be used as a gelator, as discussed
above. For instance, the amphiphilic prodrug can be contacted with
a solvent to induce self assembly to form a prodrug-based gelator.
Suitable solvents include water, water containing one or more
salts, organic solvents, and oils. Preferred organic solvents
include hexane, heptane, benzene, toluene, mesitylene,
ethylacetate, chloroform, ethanol, propanol, butanol, tertiary
butanol, tetrahydrofuran, acetonitrile, and combinations thereof.
Preferred solvent oils include petrol, diesel, gasoline, vegetable
oil, cooking oil, polyunsaturated oils, and combinations
thereof.
[0080] The functional groups of the gelator promote self assembly
by facilitating hydrogen bonding, intermolecular interactions
through pi-pi stacking, and molecular association through van der
Waals interactions, as discussed above. Self assembly may take
place in the presence of an encapsulating one or more agents so
that the agent or agents are encapsulated in the gel.
[0081] Preferably, the compound and the agents are both drugs or
prodrugs. More preferably, the compound is acetaminophen, and most
preferably, the amphiphilic prodrug is a compound selected from
formula (I) or formula (II), discussed above. The reaction
typically takes place at about 37-40.degree. C. for about 24-72
hours under continuous agitation.
[0082] In certain embodiments, the drug is indomethacin or
ethambutol. In some embodiments, indomethacin or ethambutol is
connected to a peptide, fatty acid or sugar moiety.
[0083] In some other embodiment, the indomethacin is of formula
(V).
[0084] In yet still other embodiments, the ethambutol is of formula
(VI).
[0085] Another embodiment relates to a single-step method of making
a low molecular weight, amphiphilic gelator, comprising the step of
reacting a tris-hydroxyl methyl amine, such as tris, with a fatty
acid or methyl ester of a fatty acid to form an amphiphilic gelator
having an amide functionality. Suitable fatty acids include fatty
acids having a saturated or unsaturated C.sub.1-21 alkyl group,
such as lauric, caprylic, myristic, palmitic, and oleic acid. The
reaction can take place without the use of a catalyst. Preferred
reaction conditions include having the reaction takes place in
dimethylsulfoxide and K.sub.2CO.sub.3 at about 20.degree. C. for
about 24-48 hours.
[0086] As discussed above, the gelator may be contacted with a
solvent to induce self assembly. The amphiphilic gelator having an
amide functionality is preferably of formula (IV), above.
[0087] Another embodiment relates to a single-step method of making
a low molecular weight, amphiphilic prodrug, comprising the step of
esterifying a compound having a saccharide group, phenol ring and
primary hydroxyl group with a vinyl ester of carboxylic acid to
form an amphiphilic prodrug containing an ester functionality.
[0088] The esterification reaction preferably takes place in
presence of a Lipase enzyme, such as lipolase. The reaction
typically takes place at about 37-40.degree. C. for about 24-72
hours under continuous agitation. The amphiphilic prodrug is
preferably salicin or a salicin derivative, such as that of formula
(III), above.
[0089] Salicin has generally been considered a prodrug of aspirin.
It contains a polysaccharide ring, an aromatic group and hydroxyl
group. To prepare the preferred gelator of this embodiment, a
hydrophobic polymethylene chain can be reacted with the primary
hydroxyl group of the glucose group in the salicin to make the
amphiphilic prodrug-based gelator.
Method of Drug Delivery
[0090] Another embodiment of this invention relates to a method of
drug delivery. The method contains a first step of administering to
a patient in need thereof a drug-derived or prodrug-derived gelator
capable of self assembly. Upon self assembly, the gelator becomes a
self-assembled gelator. The backbone of the gelator contains the
drug or prodrug, a hydrophobic moiety, and a degradable link
between the drug or prodrug and hydrophobic moiety. The second step
involves contacting the compound or gelator with an enzyme (in an
enzymatic cleavage) or water (in a hydrolytic cleavage) to cleave
the degradable link and release the drug or prodrug in the
patient.
[0091] Preferably, the gelator is an acetaminophen-derived gelator
or a salicin-derived gelator, as discussed above. Preferred
acetaminophen-derived gelators include gelators selected from
formula (I) or formula (II), and preferred salicin-derived gelators
include gelators having formula (III), above. The gelator may
include one or more drugs encapsulated in the gel of the
gelator.
[0092] In certain embodiments, the gelator is indomethacin or
ethambutol derived hydrogelator. In some embodiments, indomethacin
or ethambutol is connected to a peptide, fatty acid or sugar
moiety.
[0093] In some other embodiment, the indomethacin is of formula
(V).
[0094] In yet still embodiments, the ethambutol is of formula
(VI).
[0095] The degradable bond between the drug or prodrug and
hydrophobic moiety may be any bond known in the art that is
degradable enzymatically or hydrolytically. Bonds capable of being
cleaved include ester, amide, anhydride, and carbamate bonds.
[0096] When the gelator is contacted with an enzyme under
conditions sufficient to cleave the degradable bond, an enzymatic
cleavage occurs. Any enzyme capable of cleaving the bond may be
used. Suitable enzymes include Lipase enzymes, such as lipolase.
The enzyme cleaves the degradable bond in the compound to release
the derived drug or prodrug and the fatty acid.
[0097] In instances when the gelator contains one or more drugs
encapsulated in the self-assembled gel, the enzyme cleaves the
degradable bond in the gelator to release the encapsulated drugs,
the derived drug or prodrug, and a fatty acid. In this instance,
the enzyme cleavage releases the encapsulated drug and derived drug
or prodrug simultaneously or nearly simultaneously. Preferably, the
enzymatic cleavage takes place under physiological conditions.
[0098] As discussed above, the self-assembled gelator may
encapsulate one or more drug as well as one or more agents, such as
an enzyme, in the gel of the gelator. An embodiment of this
invention thus relates to a self-assembled gelator that contains an
enzyme, where the encapsulated enzyme cleaves the gelator and
releases the drugs. The encapsulated enzyme may be present within
the hydrogel during the gelation process, and then be released so
that it can cleave the gelator and subsequently release the drug or
prodrug. The enzyme may also be an external enzyme, or an enzyme
that is otherwise not encapsulated in the gelator.
[0099] When the gelator is contacted with water under conditions
sufficient to cleave the degradable bond, a hydrolytic cleavage
occurs. Similar to the enzymatic cleavage, the hydrolytic cleavage
can release the derived drug or prodrug and the fatty acid, or,
when the gelator contains one or more encapsulated drugs, the
cleavage can release the encapsulated drugs, the derived drug or
prodrug, and a fatty acid. Hydrolytic cleavage often takes place at
elevated or basic pH conditions, for instance, at a pH ranging from
about 6-12.
[0100] The drug is capable of being delivered in a sustained
release. Sustained release, as appreciated by those of skill in the
art, can delay the release of the drug or prodrug for a desired
period of time. Depending on the parameters, the release can be
delayed from minutes to days to months or even years, especially
when administered under physiological conditions (a pH of about 7.4
and a temperature of about 37.degree. C.). The sustained release
can be controlled by the concentration of the enzyme and, if
applicable, the body temperature of the patient. For instance,
sustained release can be accelerated via high enzyme concentration.
In a preferred embodiment, the sustained release is delivered
without a burst release, or with only a minimal burst release.
[0101] Another embodiment of this invention relates to a method of
drug delivery, comprising a first step of administering to a
patient in need thereof a drug or prodrug encapsulated in a
self-assembled gel. The backbone of the gelator contains an amide
functionality and a hydrophobic moiety. The second step involves
contacting the gelator with a suitable enzyme to cleave the amide
bond in the gelator and release the drug or prodrug in the
patient.
[0102] Other degradable links besides the amide bond may also be
used. In this embodiment, the method of drug delivery contains a
first step of administering to a patient in need thereof a drug or
prodrug encapsulated in a self-assembled gelator. The backbone of
the gelator contains a hydrophobic moiety and a degradable link
between the drug or prodrug and the hydrocarbon chain. The second
step involves contacting the gelator with a suitable enzyme to
cleave the degradable link in the gelator and release the drug or
prodrug in the patient. The cleavage takes place under
physiological conditions.
[0103] Preferably, the gelator is a tris-based amphiphile, for
instance the reaction product of
2-amino-2-hydroxymethyl-propane-1,3-diol and a fatty acid or methyl
ester. Preferred gelators are of formula (IV), discussed above.
[0104] The drugs and prodrugs, whether encapsulated and/or part of
the backbone of the gelator, can be administered to a patient that
is suffering from an ailment or otherwise in need of such drugs.
Typical ailments include as cancer, arthritis, cardiovascular
disease, and the like. Materials made from, for instance, salicin
or salicin derivatives and acetaminophen or acetaminophen
derivatives can be used as anti-inflammatory materials. These drugs
and prodrugs can be combined other anti-inflammatory drugs or their
prodrugs to treat, for example, various arthritis ailments.
[0105] It is possible to administer the drugs or prodrugs through
various known delivery techniques, including injection and
implantation. Injection and implantation are particularly feasible
in view of the ability of the gelator to form in situ (i.e. in situ
self-assembly). Injecting or implanting the drug or prodrug
composition into a joint together with the sustained-release
properties enables the drug to provide a long-term release over a
period of time. This is particularly suitable in instances where
enzymes that are present in a joint are naturally released upon
inflammation of the joint. When the joint becomes inflamed and
releases the enzyme, the enzyme, in turn, cleaves the drug or
prodrug composition, which releases the anti-inflammatory drugs.
After the anti-inflammatory drug is released, the enzyme
concentration decreases. The drug or prodrug composition that is
not cleaved remains stable until another inflammatory stimulus.
This phenomenon can be referred to as "on-demand release," where
the level of inflammation regulates the amount and timing of
drug/prodrug release.
Compounds
[0106] It is believed the compounds discussed above as gelators are
novel in their own right. Therefore, an additional embodiment of
this invention is directed to the following individual compounds,
separate from their use as gelators.
[0107] One embodiment thus relates to a compound of formula
(I):
##STR00006##
where n represents an integer from 1-22, preferably ranging from
1-12, and more preferably from 6-12.
[0108] Another embodiment relates to a compound of formula
(II):
##STR00007##
where n represents an integer from 1-22, preferably ranging from
1-12, and more preferably from 6-12.
[0109] Another embodiment relates to a compound of formula
(III):
##STR00008##
where R is a saturated or unsaturated C.sub.1-21 alkyl group.
[0110] Another embodiment relates to a compound of formula V:
##STR00009##
wherein R is peptide comprising 1-40 amino acids or R is saturated
or unsaturated C.sub.1-C.sub.21 alkyl group. In some embodiments, R
is C.sub.1-C.sub.6 alkyl. In some other embodiments, R is
C.sub.6-C.sub.12 alkyl.
[0111] Another embodiment relates to a compound of formula VI:
##STR00010##
wherein R is a peptide comprising 1-40 amino acids or R is
saturated or unsaturated C.sub.1-21 alkyl group. In some
embodiments, R is C.sub.1-C.sub.6 alkyl. In some other embodiments,
R is C.sub.6-C.sub.12 alkyl.
EXAMPLES
[0112] Although preferred embodiments have been depicted and
described in detail herein, it will be apparent to those skilled in
the relevant art that various modifications, additions,
substitutions, and the like can be made without departing from the
spirit of the invention and these are therefore considered to be
within the scope of the invention as defined in the claims which
follow.
Example 1: Self-Assembled Acetaminophen Prodrugs
[0113] Utilization of enzyme catalysis as a tool to disassemble
self-assembled hydrogels to control the release encapsulated drug
provides an opportunity to employ a wide range of enzyme-specific
low-molecular-weight hydrogelators.
[0114] In this example, low-molecular-weight amphiphilic prodrugs
are synthesized as hydrogelators from biocompatible fatty acids and
a well-known drug, acetaminophen (Apn), (which belongs to a class
of drugs called analgesics (pain relievers) and antipyretics (fever
reducers)). This reaction was performed in a single-step
esterification. The example shows the prodrug's ability to
self-assemble into nanoscale structures in aqueous solutions to
form hydrogels that may subsequently encapsulate a second drug such
as curcumin, which is a known chemopreventive and anti-inflammatory
hydrophobic drug. Upon enzyme triggered degradation, the hydrogel
can release single or multiple drugs at physiologically simulated
conditions in vitro.
[0115] Given that the degradation products consist of the drug and
a fatty acid, this approach has an advantage over polymer-based
prodrugs that generate polymer fragments with heterogeneous chain
lengths upon degradation that may present complex toxicity
profiles. Additionally, drug release can occur without burst
release by an enzyme triggered hydrogel degradation mechanism.
Spectrophotometric experiments supported the drug release, and the
rate was controlled by modulation of temperature and enzyme
concentration. To test the cytocompatibility of amphiphilic
prodrugs, and their effect on cell characteristics such as
viability, proliferation, adhesion and cell phenotype, a series of
experiments was performed with mesenchymal stem cells (MSCs). After
treating the MSCs with amphiphilic prodrugs, it was found that the
MSCs retain their stem cell properties, including the capacity of
multi-lineage differentiation, and maintain their adhesive and
proliferation capacities with high viability. Therefore, it is
believed that this approach has broad applications as drug delivery
vehicles from a wide range of prodrug-based gelators.
[0116] The existing ambiguity can be substantially decreased by
designing prodrugs-based LMWGs from existing drugs, for instance
existing drugs whose metabolic pathways are well studied. Such
prodrugs may also encompass functional groups which can promote
self-assembly in aqueous solutions, and pose ability to encapsulate
drugs within the pockets of hydrogel.
[0117] In this example, delivery vehicles are developed for single
and multiple drugs. Upon degradation, drug-encapsulated
prodrug-based hydrogels can release drug-1 (which generates from
cleavage of prodrug) and drug-2 (the encapsulated drug(s)). See the
FIG. 1 for schematic representation of single and multiple drug
delivery from prodrug based hydrogels.
[0118] Drug delivery can occur from hydrogels through various
mechanisms such as swelling and dissolution so on. Biodegradability
or dissolution may be designed into hydrogels via enzymatic,
hydrolytic, or environmental (e.g. pH, temperature, or electric
field) pathways. Enzymes, which can permit the release of
encapsulated therapeutics in desired locations, can be employed as
stimuli for drug release, for instance, in tumors as a result of
the enzymatic action of tumor-associated proteases such as plasmin,
or in selected sites of the gastro intestinal tract under the
influence of digestive enzymes. Enzymes have also been used in
biocatalysis as a tool to make the gelators from biomass and as
stimuli to degradation of the gel which triggered drug
delivery.
[0119] In this example, biocompatible amphiphilic prodrugs as LMWGs
have been developed to encapsulate hydrophobic drugs and
subsequently release single and multiple drugs upon enzyme mediated
gel degradation (FIG. 1). The well-known drug
N-(4-hydroxyphenyl)acetamide, which is also known as acetaminophen
(Apn), was chosen, as its drug activity has been well studied. Apn
is a common analgesic and antipyretic drug that is used for the
relief of fever, headaches, and other minor aches and pains. It is
remarkably safe in recommended doses, and its route of metabolism
in human has extensively been investigated and found to be
safe.
[0120] A series of amphiphiles and bolaamphiphiles were synthesized
in a single step from Apn as shown in FIG. 2. The designed Apn
based prodrugs encompass functional groups which can promote
intermolecular molecular interactions through hydrogen bonding,
pi-pi stacking and van der Waals interactions.
[0121] Design and Synthesis of Prodrug-Based Gelators. Design of
amphiphilic gelators from known drugs is a challenging task. During
this process the following parameters were used: i) the drug
molecule had a functional group which can be easily functionalized
under mild conditions; ii) the functionalized drug (prodrug) was
able to undergo defunctionalization (reversible) under `triggered`
conditions to release the parent drug; iii) the drug has the
ability to promote self-assembly, or introducing appropriate
additional groups. While small molecular drugs have previously been
combined with polymers (see Huang, X.; Brazel, C. S. J. Controlled
Release 2001, 73, 121-136). or polymeric micelles (see Hans, M.;
Shimoni, K.; Danino, D.; Siegel, S. J.; Lowman, A.
Biomacromolecules 2005, 6, 2708-2717; and Kim, S. C.; Kim, D. W.;
Shim, Y. H.; Bang, J. S.; Oh, H. S.; Wan-Kim, S.; Seo, M. H. J.
Controlled Release 2001, 72, 191-202), it is believed that there
are no reports on developing single and multiple drug delivery
vehicles from self-assembled prodrugs. Hence, Apn was chosen for
generating prodrug based hydrogelators.
[0122] Apn encompass acetamide group which can promote hydrogen
bonding while phenyl ring can facilitate intermolecular
associations through pi-pi stacking. However it is also water
soluble. Thus, the addition of a biocompatible fatty acid through a
labile ester linker (Apn-11-CH.sub.3) will insert the ability to
form van der Waals interactions into amphiphile. To improve the
polarity for dispersion into water upon heating, bolaamphiphiles
were designed. See Scheme in FIG. 2. Introducing a terminal
carboxyl group (Apn-7-COOH and Apn-13-COOH) renders multiple
advantages such as increasing the polarity, additional hydrogen
bond forming ability through carboxylic groups and the possibility
to better control the self-assembly through pH variation.
Symmetrical bolaamphiphiles were synthesized connecting two Apn
molecules through a dicarboxylic acid linker (Apn-8-Apn and
Apn-14-Apn). In addition, to probe the mechanistic properties of
enzyme triggered gel degradation and drug delivery, a hydrocarbon
chain was coupled through a non-hydrolyzable ether linker
(Apn-5-ether-COOH, Scheme in FIG. 2), which also has a terminal
carboxylic group. Hence it can retain its self-assembling ability.
Apn based amphiphiles and bolaamphiphiles were synthesized using
synthetic techniques in a single step, as shown in FIG. 2 and the
Experimental section, below. It is believed that this synthetic
route can be used to develop these gelators in industrial scales
for various applications.
[0123] Gelation Studies with Prodrug-Based Gelators. Gelation
abilities of Apn based amphiphilic prodrugs have been investigated
thoroughly in a wide range of solvents. For detailed gelation
results, see Table 1, below). Typically, the required amount of
gelator was placed in a glass vial and with an appropriate solvent.
Subsequent heating resulted in the dissolution of the gelator. The
resulting solution was slowly cooled to room temperature and
gelation was visually observed. Gelation was confirmed when no
gravitational flow was observed in an inverted vial. All Apn based
amphiphiles were observed to be good thermoreversible gelators.
Amphiphile Apn-11-CH.sub.3 did not gelate in water while gelating
various organic solvents. Despite of prolonged heating
Apn-11-CH.sub.3 was not soluble in water. In contrast, Apn-7-COOH
forms excellent gels in water and other aqueous buffer solutions
even in presence of salt. In addition, Apn-7-COOH is soluble in
basic solutions (pH is more than 8). This shows that terminal
carboxylic acid groups play a role in self-assembly, possibly
through formation of hydrogen bonding network. However, due to
longer hydrophobic chain Apn-13-COOH was sparingly soluble in water
and thus additional co-solvent (5-15 v/v % of alcohol such as
methanol, ethanol or isopropanol) was used to make the hydrogel.
Symmetrical bolaamphiphile Apn-8-Apn formed stable hydrogels while
Apn-14-Apn was insoluble in water.
[0124] These results show that a balance of solvophobic and
solvophilic is desirable while other functional groups promote
intermolecular interactions such as hydrogen bonding, pi-pi
stacking and van der Waals interactions. All gelators showed lower
minimum gelation concentrations (between 0.4-1.5 w/v % and
millimolar concentrations, Table S1). Gel to solution transition
temperature (T.sub.gel) was determined by typical
inversion-tube-method (see, for example Vemula, P. K.; John, G.
Chem. Commun. 2006, 2218-2220, and F. M.; Caran, K. L. J. Am. Chem.
Soc. 2000, 122, 11679-11691) and they exhibited values between
60-82.degree. C. for 1 wt % of gels depending on the solvent used.
All gels were opaque in nature and stable for several weeks. These
results indicate that prodrug based hydrogels exhibit high thermal
and temporal stabilities. Detailed gelation results are shown in
Table 1.
[0125] Hydrogel Morphology. The morphologies of self-assembled
hydrogels were examined under scanning and transmission electron
microscopy (SEM and TEM, respectively). Investigation of the
hydrogels of Apn-7-COOH and Apn-8-Apn with SEM and TEM showed that
prodrug gels form branched or entangled fibrous/sheet-like gel
networks with fiber thickness of 50-400 nm, and fiber lengths of
several microns (FIGS. 3A-3E). The high aspect ratios of the gel
fibers indicate that the intermolecular interactions between the
gelator molecules are highly anisotropic. In addition, lower
gelation concentrations and high temporal stability of the gels
suggest that the intermolecular interactions are more prevalent and
thus most likely the result of the synergistic effect of both pi-pi
interactions and hydrogen bonding.
[0126] Self-Assembly in Hydrogels. Optimized geometries were
obtained and the lengths of gelators were calculated using ab
initio calculations. By combining the data from X-ray diffraction
(XRD) and Fourier transform infrared spectroscopy (FT-IR) a model
for self-assembly of amphiphilic prodrugs in aqueous solution was
proposed. All the ab initio Hartree-Fock calculations reported were
performed using the Gaussian 03 suite program (Frisch, M. J. et al.
Gaussian 03, Revision B.03; Gaussian, In.: Pittsburgh, Pa., 2003).
The geometries of Apn-7-COOH and Apn-8-Apn have been located and
optimized at the level of restricted Hartree-Fock (RHF) using the
6-31 G* basis set (Hehre, W. J.; Radom, L.; Schleyer, P. V. R. Ab
Initio Molecular Orbital Theory; Wiley & Sons: New York 1986,
pages 63-91. All structures were completely optimized without any
symmetry restrictions. For both molecules studied, vibrational
frequency calculations were carried out to confirm that they
converge to true minima by diagonalization of their Hessian (force
constant) matrixes at the same level and making sure that all
frequencies are real. To check the reliability of the level of
theory, Apn was optimized using RHF/6-31 G*, thus geometrical
parameters were compared with the reported crystal structure of Apn
(FIG. 15, Table 2). Fully optimized geometries of both amphiphilic
prodrugs are shown in FIG. 4A, and calculated lengths are 1.97 and
2.67 nm for Apn-7-COOH and Apn-8-Apn, respectively.
[0127] The hydrogels obtained from prodrugs Apn-7-COOH and
Apn-8-Apn displayed well-resolved X-ray diffraction (XRD) patterns
that were characteristic of the long-range ordering of the
gelators. The long spacing (d) was calculated to postulate the
possible mode of self-assembly in the gel state. Long distance
spacing values suggest that possibly lamellar structures were
formed by these gelators in the gels. In XRD measurements, the
hydrogel of Apn-7-COOH a d of 3.53 nm, which is higher than the
molecular length of Apn-7-COOH (1.97 nm from the optimized geometry
calculations, FIG. 4A) and slightly lower than double the molecular
length. Therefore, it can be predicted that amphiphiles are tilting
with respect to normal to the layer plane (FIG. 4B).
[0128] The hydrogel of Apn-8-Apn showed two major d values of 2.72
and 4.9 nm. In the case of d=2.72 which is close to the molecular
length of Apn-8-Apn, it is believed that a monolayer-like structure
exists which could be stabilized by strong intermolecular
interactions such as hydrogen-bonding, pi-pi interactions, as shown
FIG. 4C. Additionally, acetaminophen groups of two adjacent layers
could form strong hydrogen-bonding as shown in FIG. 4D which
supports the observed d value 4.9 nm. In addition, FT-IR
experiments were performed to gain insight into the
hydrogen-bonding environment of the amide carbonyl (C.dbd.O) group
in Apn-8-Apn under solution and gel conditions. In the methanolic
solution of Apn-8-Apn, an amide I band appeared at 1663 cm.sup.-1
whereas in gel state it shifted to 1631 cm.sup.-1. Such a
significant shift of 32 cm.sup.-1 could be attributed to the strong
hydrogen-bonding interactions of the carbonyl groups of acetamides
in the gel state which suggests that hydrogen-bonding enhances the
self-assembly of these amphiphilic prodrugs to form hydrogels.
Thus, based on the information obtained from the theoretical
calculations, FT-IR and XRD experimental results, a model for the
self-assembly of these amphiphiles has been proposed, as shown in
FIGS. 4A-4D.
[0129] Enzyme-Mediated Single and Multiple Drug Release. Many
potent therapeutic agents possess a high degree of hydrophobicity
which can impede their solubilization in aqueous media and thus
hamper their oral or parenteral administration. In order to
circumvent this limitation, suitable biocompatible drug delivery
systems can be developed which can provide increased surface area
for it to dissolve by providing many hydrophobic drug molecules
throughout the assembled gel. In this example, the encapsulation of
hydrophobic drug in a prodrug-based hydrogel is demonstrated, which
upon enzyme-mediated gel degradation releases single and multiple
drugs at physiological conditions in a controlled manner.
[0130] To investigate single and multiple drug delivery from
Apn-based prodrug hydrogels, the well-known chemopreventive drug
curcumin was chosen as a model hydrophobic drug for encapsulating
into hydrogels. Curcumin (1,7-bis(4-hydroxy-3
methoxyphenyl)-1,6-heptadiene-3,5-dione) was originally extracted
from the root of Curcuma longa, and it is the main constituent of
the Indian spice turmeric, and give curry sauces their
characteristic yellow color. This molecule shows a wide range of
pharmaceutical activity, including potent antioxidant,
anti-inflammatory, antiseptic and anti-carcinogenic properties. It
also exhibits promising therapeutic activity by inhibiting various
purified human immunodeficiency virus (HIV) types.
[0131] However, in spite of promising drug activity, curcumin has
an extremely poor bioavailability due to particularly its low
solubility in aqueous solutions (solubility of curcumin in water is
3.times.10.sup.-8 M). Previously, Tang et al. demonstrated
increased solubility of curcumin in water by forming supramolecular
host-guest complex with various cyclodextrins (Tang, B.; Ma, L.;
Wang, H.-Y.; Zhang, G.-Y. J. Agric. Food Chem. 2002, 50,
1355-1365). Hence, it would be possible to increase curcumin
concentration in water by encapsulating into hydrogels where
abundant hydrophobic regions are available. Hydrogels of Apn-7-COOH
or Apn-8-Apn could encapsulate high concentrations of curcumin (up
to 0.5 mmol); the observed enhanced concentration is believed to be
due to localization of hydrophobic curcumin within the hydrophobic
pockets of hydrogels three-dimensional network.
[0132] To examine the effect of external drug encapsulation on the
inherent morphology of the self-assembled gel,
curcumin-encapsulated hydrogels were examined under SEM and TEM
(FIGS. 5A-5E). Encapsulation of curcumin did not affect the
morphology of the hydrogels. It is believed that self-assembly
phenomenon could be identical or nearly identical in both types of
hydrogels (with and without curcumin), as there was no significant
difference in the morphology of the native hydrogels and curcumin
encapsulated hydrogels (see FIGS. 3A-3E and FIGS. 5A-5E,
respectively).
[0133] Encapsulation and release of curcumin was previously studied
by the Ultra-Violet absorbance spectroscopy (.lamda.max at 424 nm).
Absorbance spectrum of modified Apn-7-COOH was similar to that of
Apn (FIG. 6B). In this instance, enzyme-triggered single and
multiple drug delivery has been carried out using two sets of
hydrogel samples, i.e., hydrogels of Apn-7-COOH and Apn-8-Apn with
and without curcumin. In the first set, lipase (Lipolase 100 L,
Type EX, lipase units 100 KLU/g) was added to the preformed
hydrogel of Apn-7-COOH (2 wt %) and kept it at 37.degree. C. which
is lower than gel melting temperature. It was anticipated that
lipase (esterase) would hydrolyze the ester bond of prodrug-based
gelator (Apn-7-COOH) to release acetaminophen and dicarboxylic acid
as the hydrogel degrades. As expected, after 48 hours the gel
completely degraded and became a clear solution. UV-absorption
experiments could not study the release of original drug
acetaminophen (Apn), as absorbance peaks of Apn and lipase overlap
significantly. Hence, thin-layer chromatography was performed on
resulted clear solution, which indicated that indeed acetaminophen
and .alpha.,.omega.-dicarboxylicacid were generated in the
solution. This suggests that the gel degradation is occurring
through the cleavage of ester bonds within the acetaminophen-based
prodrugs by lipolase enzyme.
[0134] To test this further, and examine multiple (two) drugs
delivery (FIGS. 6A-6E), curcumin was encapsulated in the hydrogel
of Apn-7-COOH (2 wt %) and to that lipase was added and kept it at
37.degree. C. Absorbance spectra were recorded on aliquots which
were collected (after the addition of enzyme to the hydrogel) at
regular intervals (FIG. 6C). Plotting absorbance maxima vs time
(FIG. 6E) reveals absence of burst release which is otherwise a
routine hurdle in hydrogel-based drug release. Initial aliquots did
not show any absorbance peak, but aliquots collected after 4 hrs
showed absorption maxima at 425 nm, which corresponds to the
absorption peak of curcumin. The concentration of the enzyme used
in these in vitro experiments is higher than physiological
concentration of esterases (for example, physiological
concentration of cholesterol esterase is 7 U/ml). Hence it was
anticipated that under in vivo conditions the present system would
show prolonged sustained release.
[0135] To determine the role of the enzyme on hydrogel degradation,
similar experiments were performed by adding only buffered solution
without an enzyme. As expected, curcumin encapsulated gel was still
intact over a month, there was no visual change in the gel volume
and added solution, and no absorbance peaks corresponding to the
curcumin were observed, suggesting that the enzyme is effective in
the gel degradation. In addition, it also suggests that the
curcumin release which was observed in the previous experiment is
due to degradation of hydrogel. There was no loosely absorbed
curcumin on hydrogels to diffuse into the water with time. Hence,
it is believed that the encapsulated curcumin intercalates within
in the hydrophobic pockets of self-assembled fibers.
[0136] The yellow color cleared solution which was obtained after
enzyme mediated gel degration was also tested by thin-layer
chromatography, and found that Apn and curcumin are present; thus,
it confirms the synergistic release of Apn and curcumin which
indicate that this approach could be used to develop delivery
vehicles to release two drugs (possible complimentary drugs)
simultaneously or nearly simultaneously. Increasing either
concentration of enzyme or incubation temperature to 45.degree. C.
doubles the drug release rate, which was correlating with previous
findings.
[0137] To analyze the gel degradation mechanism, another prodrug
Apn-5-ether-COOH was synthesized (FIG. 2), where hydrocarbon chain
(which is bearing terminal carboxylic acid) was connected to the
acetaminophen through an ether bond (C--O--C) to prevent the
esterolytic cleavage by lipase. Apn-5-ether-COOH exhibited gelation
behavior akin to Apn-7-COOH; curcumin has been encapsulated into
Apn-5-ether-COOH hydrogel, and to that lipase was added. After
incubating at 37.degree. C. for several days, the hydrogel was
still intact and prevented curcumin release into the solution
(confirmed by absorption spectroscopy) which suggests that the
lipase cleaves the prodrug gelator at the ester bond to release Apn
and encapsulated curcumin. Hence, having a cleavage labile ester
group is desirable when preparing the prodrug based
hydrogelators.
[0138] Cytocompatibility. The cytocompatibility of prodrugs
generated from small modification of known drugs was investigated.
This was done to demonstrate that chemical modification on Apn did
not introduce cytotoxicity into the molecule. To test the
cytocompatibility of prodrug-based amphiphiles and bolaamphiphiles,
and their effect on cell characteristics such as viability,
proliferation, adhesion and cell phenotype, a series of experiments
was performed with mesenchymal stem cells (MSCs). To gain more
insight into the effect of modification, unmodified drug Apn was
used as a control. The viability of control cells and cells treated
with Apn, Apn-7-COOH and Apn-8-Apn was examined with trypan blue
assay. Use of Mesenchymal stem cells is appropriate given that
these systems may be used for localized release likely into
connective tissues.
[0139] The results (FIG. 7A) suggest that there was no significant
decrease in the viability of the MSCs after 48 hours incubation
with prodrug-based amphiphiles. Specifically, 95% of untreated
cells were viable after 48 hours, whereas 91%, 87% and 82% of
viable cells were observed for cells incubated with 250 .mu.M of
Apn, Apn-7-COOH and Apn-8-Apn, respectively (FIG. 7A).
[0140] In another set of experiments, it was determined that
viability rates were not significantly affected by a wide range of
concentration of Apn-7-COOH (up to 500 .mu.M, FIG. 7B). These
results show that Apn-derived prodrug amphiphiles did not induce
cell toxicity. To test how prodrug hydrogelators will affect the
extracellular environment of MSCs, cell adhesion characteristics
were examined (after 12 hrs), and it was found that in presence of
Apn-based amphiphiles there was no considerable change in adhesion
properties of MSCs (FIG. 7C).
[0141] The proliferation rate of MSCs was also examined. In all
cases (cells treated with buffer and cells treated with Apn,
Apn-7-COOH and Apn-8-Apn) cells proliferated to attain a confluent
layer, and were found to have similar proliferation rates (FIG.
7D). This indicates that the presence of prodrug-amphiphiles did
not alter the proliferation of MSCs.
[0142] Ethanol was used to dissolve amphiphiles, with the total
volume of the ethanol being 1 v/v % in the experiments. To exclude
the possibility that observed small changes in viability, adhesion
and proliferation results was caused by ethanol, in every
experiment, the control was used where cells were treated with 1
v/v % of ethanol. The results (not shown here) demonstrate that
ethanol did not significantly affect the viability, adhesion and
proliferation.
[0143] Unlike typical fibroblast cell lines used for toxicity
screening, cultured mesenchymal stem cells are highly sensitive to
the presence of soluble factors and can often loose their capacity
for multi-lineage differentiation or differentiate into non-desired
lineages depending on the culture conditions. To examine the impact
of the acetaminophen amphiphilic prodrugs on multi-lineage
differentiation of MSCs, cells were treated with adipogenic and
osteogenic induction media in presence of amphiphiles followed by
respective colorimetric histological staining. Even in presence of
Apn, Apn-7-COOH and Apn-8-Apn, the MSCs showed a robust capacity
differentiated into adipogenic (FIGS. 8A-8E) and osteogenic (FIGS.
9A-9E) lineages as shown by oil red O (ORO) staining and alkaline
phosphatase (ALP) activity, respectively. No significant difference
in ORO or ALP staining was observed for MSCs treated with and
without Apn-based prodrug amphiphiles. Thus, the modified prodrugs
did not impair the potential for multi-lineage differentiation.
Hence, the Apn-based prodrugs are cytocompatible and do not amend
the inherent cellular properties of MSCs.
[0144] Example 1 demonstrates that amphiphilic low-molecular-weight
prodrug hydrogelators have been synthesized from simple
modification of known-drug acetaminophen by combining a
biocompatible fatty acid. The acetaminophen-based prodrugs
self-assembled to form hydrogels. A known chemopreventive
hydrophobic drug curcumin has been encapsulated into the hydrogels,
subsequently; enzyme-triggered single and multiple drug (one and
two drugs) delivery had been demonstrated at physiological
conditions in vitro. See FIGS. 10A-10C.
[0145] These prodrugs exhibited excellent self-assembling
properties to form hydrogels at low concentrations. Mode of
self-assembly in hydrogels was proposed based on the XRD, FTIR data
and theoretical calculations. The encapsulation of chemopreventive
curcumin in the hydrogels which were prepared from Apn-based
prodrugs was shown, and enzyme-triggered gel degradation was
performed to achieve single and multiple drug delivery in vitro at
physiological conditions. In addition, it was found that the
modified Apn-based prodrugs are excellent cytocompatible
amphiphiles, and do not alter mesenchymal stem cell properties such
as viability, adhesion, proliferation and ability to differentiate
into multiple lineages including adipogenic and osteogenic. These
results suggest that self-assembled hydrogelators from existing
drugs which have known metabolic pathway can be used in the field
of low-molecular-weight hydrogelators based drug delivery.
Experimental Section
[0146] General Information. Acetaminophen and curcumin were
purchased from Acros Chemicals (Fisher Scientific Company, Suwane,
Ga.). The Novozyme 435 [lipase B from Candida antarctica, (CALB)]
and Lipolase 100 L was obtained from Novozymes through Brenntag
North America. Human MSCs were obtained from the Center for Gene
Therapy at Tulane University. .alpha.-MEM, Fetal Bovine Serum,
L-Glutamine and Penn-Strep were purchased from Invitrogen. All
other chemicals and reagents were purchased from Sigma Aldrich (St.
Louis, Mo.) and were used without further purification unless
specified.
[0147] Synthesis of Hydrogelators. For detailed synthesis
procedures and characterization of compounds see Supporting
Information, below.
[0148] Preparation of Gels. Typically, gelator (0.1-5 mg) and
required solvent (0.1-1 mL) were placed into a 2 mL scintillation
vial, which was then sealed with a screw cap. The vial was heated
with shaking until the solid was completely dissolved. Gelation was
considered to be occurred when no gravitational flow was observed
in the inverted tube, when vial reached to the room
temperature.
[0149] Gel Melting Temperatures. Gel to Sol transition temperature
(T.sub.gel) was determined typical `inversion tube method` (see, P.
K.; John, G. Chem. Commun. 2006, 2218-2220, and Menger, F. M.;
Caran, K. L. J. Am. Chem. Soc. 2000, 122, 11679-11691) where gel
was prepared in a 2 mL glass vial by dissolving 1 wt % gelator in
required amount of solvent and closed with tight screw cap. The
vial was immersed in the water `up side down` and slowly heated,
where the viscous gel has melted dropped down that temperature was
considered as T.sub.gel.
[0150] Scanning Electron Microscopy. To record SEM, the xerogel
samples were prepared by freezing-and-pumping method from their gel
phases below the sol-gel transition temperature. The SEM images of
xerogels and the following drying under ambient condition show
similar morphologies. Therefore, morphology with the gels was
studied dried under ambient conditions, (called xerogels).
[0151] Transmission Electron Microscopy. TEM was recorded by using
Zeiss EM 902 transmission electron microscope (80 kV). A small
portion of gel was drop cast on a Cu-grid. After drying the grid at
ambient temperature, it was directly imaged under TEM.
[0152] UV-vis Spectroscopy. UV-visible spectra of the amphiphiles
and curcumin were were recorded using CARY100BIO spectrophotometer.
In all experiments, solutions were taken in quartz cuvette of 1-cm
path length.
[0153] X-ray Diffraction (XRD). XRD measurements were conducted
using a Bruker AXS D-8 Discover with GADDS diffractometer using
graded d-space elliptical side-by-side multilayer optics,
monochromated Cu-K.alpha. radiation (40 kV, 40 mA), and imaging
plate.
[0154] Mesenchymal Stem Cell Culture. Primary human mesenchymal
stem cells were maintained in expansion media that consisted of 15%
Fetal Bovine Serum (selected for its ability to expand MSCs), 1%
(v/v) L-Glutamine, 1% (v/v) Penn-Strep, and .alpha.-MEM. All
experiments were performed using MSCs at passage number 4-6.
[0155] Viability, Adhesion and Proliferation Characteristics. The
viability of the cells was examined using trypan blue exclusion.
Cells were plated into 12 well plates, to that 250 .mu.M of Apn or
Apn-based prodrugs were added, and incubated for 48 hours at
37.degree. C. and 5% CO.sub.2. The media was then aspirated and the
cells were detached with 200 .mu.L of cell dissociated solution
(similar quantities of floating cells were observed between the
groups, typically 5-10 per well). 300 .mu.L of media was added and
the total 500 .mu.L of the cell dispersion was collected in an
eppendorf tubes. From this 10 .mu.L of cell dispersion was diluted
to 1:1 with 4% trypan blue solution and cells were counted in a
hemocytometer to determine the number viable (non-blue) and
nonviable (blue) cells. A control for this experiment included
cells treated with only phosphate buffer saline (PBS) without an
amphiphile. Cell adhesion was quantified by measuring the number of
adherent cells on tissue culture surfaces in 96 well plates after
incubating with amphiphiles for 12 hours. For proliferation
studies, 5000 cells were added to each well of a 96 well plate in
200 .mu.L of MSC cell expansion media, and appropriate amphiphiles
were then added and incubated at 37.degree. C. and 5% CO.sub.2.
Proliferation was quantified by manually tabulating the number of
cells within the flask at multiple time points (1, 2, 3, 4, 6 and 8
days) with light microscopy at 10.times..
[0156] Osteogenic Differentiation. MSCs were seeded in the wells of
24 well plates, and were culture in MSC expansion media until they
reach 90% confluence (note: in control samples there was no
amphiphile present, whereas in experimental samples 250 .mu.M of
Apn-based amphiphile was present). Osteogenic differentiation was
induced by culturing the cells for 23 days in osteogenic induction
media (from Lonza-MSCs Osteogenic Single Quote kit) containing
dexamethasone, 0-glycerophosphate, L-ascorbic acid-2-phosphate, and
.alpha.-MEM. The media in both groups was changed every 3 days
(note: each time while changing the media, 250 .mu.M of Apn-based
amphiphiles were added to maintain constant concentration).
Osteogenesis was evaluated by cell membrane associated alkaline
phosphatase activity. Alkaline phosphatase activity was examined
after 23 days by carefully aspirating the media and washing the
cells with PBS. The cells were fixed with 3.7% formaldehyde
solution for 15 mn at room temperature followed by rinsing with
distilled water. 0.06% Red Violet LB salt solution in Tris HCl was
added with distilled water containing DMF and Naphthol AS
MX-PO.sub.4. The plates were incubated for 45 minutes at room
temperature and then the wells were rinsed 3 times with distilled
water and imaged under with inverted phase contrast microscope.
[0157] Adipogenic Differentiation. MSCs were seeded in the wells of
24 well plates, and were culture in MSC expansion media until they
reach 90% confluence (note: in control samples there was no
amphiphile present, whereas in experimental samples 250 .mu.M of
Apn-based amphiphile was present). Adipogenic differentiation was
induced by culturing the cells for 23 days in adipogenic induction
media (from Lonza-MSCs Adipogenic Single Quote kit containing
h-Insulin (recombinant), L-Glutamine, Dexamethasone, Indomethacin,
IBMX (3-isobuty-1-methyl-xanthine), Pen/Strep) and adipogenic
maintenance media (from Lonza-MSCs Adipogenic Single Quote kit
containing h-Insulin (recombinant), L-Glutamine, Pen/Strep). The
media in both groups were changed every 3 days in a periodic
exposure of induction and maintenance media as suggested by the
supplier (Lonza), (note: each time while changing the media,
another 250 .mu.M of Apn-based amphiphiles were added to maintain
constant concentration). Adipogenesis was evaluated by Oil Red O
staining. Following aspiration of the media, cells were washed once
with PBS, and fixed in PBS with 3.7% formaldehyde for 30 min at
room temperature. Following fixation cells were rinsed with
distilled water for a few minutes, and incubated with isopropanol
containing an Oil Red O working solution (prepared by diluting 30
mL of 0.5% isopropanol/Oil Red O solution with 20 mL of distilled
water). After 45 minutes culture well were rinsed twice with
distilled water and 1 mL of hematoxylin was added to each well for
1 minute followed by aspiration and washing with distilled water.
The wells were viewed using an inverted phase contrast
microscope.
TABLE-US-00001 TABLE 1 Gelation ability of acetaminophen-based
prodrug hydrogelators..sup.a Apn-11- Apn-7- Apn-13- Apn-8- Apn-14-
solvent CH.sub.3 COOH COOH Apn Apn water I G (0.4) G (0.6) G (1.1)
G (0.9) methanol S S S S S ethanol G (1.5) S S S S isopropanol G
(0.8) I S S S t-butanol G (0.6) I S S S ethyl acetate P I I I I
chloroform S S S S S .sup.aValues in the parenthesis are minimum
gelation concentration (wt/v %). G = gel, P = precipitate, S =
soluble, I = insoluble.
TABLE-US-00002 TABLE 2 Selected geometric parameters (bond lengths,
.ANG.) of Apn obtained from ab initio calculation and compared with
reported crystal structure. Inset optimized geometry of Apn using
restricted Hartree Fock level theory and 6-31G* basis set. Atom
numbering of Apn is shown in FIG. 15. crystal structure.sup.a
RHF/6-31G* O1--C1 1.375 1.355 C1--C2 1.396 1.382 C2--C3 1.385 1.385
C3--C4 1.396 1.386 C4--C5 1.396 1.393 C5--C6 1.388 1.381 C6--C1
1.389 1.385 C4--N1 1.426 1.413 N1--C7 1.346 1.361 C7--O2 1.239
1.197 C7--C8 1.509 1.516 O1--H1 0.92 0.94 N1--H2 0.92 0.99
.sup.aNaumov, D. Y.; Vasilchenko, M. A.; Howard, J. A. K. Acta
Cryst. 1998, C54, 653-655.
TABLE-US-00003 TABLE 3 Selected geometric parameters (bond angles,
degree) of Apn obtained from ab initio calculation and compared
with reported crystal structure. Inset optimized geometry of Apn
using restricted Hartree Fock level theory and 6-31G* basis set.
Atom numbering of Apn is shown in FIG. 15. crystal structure.sup.a
RHF/6-31G* H1--O1--C1 111.0 110.65 O1--C1--C2 122.13 122.94
C1--C2--C3 119.9 119.94 C2--C3--C4 120.89 120.19 C3--C4--C5 118.98
118.61 C4--C5--C6 120.09 120.08 C5--C6--C1 120.72 120.96 C6--C1--O1
118.47 117.85 C6--C1--C2 119.40 119.19 C4--N1--C7 128.41 129.18
C4--N1--H2 116.0 114.76 H2--N1--C7 115.4 116.05 N1--C7--O2 123.2
124.36 N1--C7--C8 115.11 113.64 O2--C7--C8 121.72 121.99
.sup.aNaumov, D. Y.; Vasilchenko, M. A.; Howard, J. A. K. Acta
Cryst. 1998, C54, 653-655.
Synthesis and Characterization of Acetaminophen Based
Amphiphiles.
[0158] General procedure for synthesis of Apn-7-COOH and
Apn-13-COOH. In a round bottom flask, .alpha.,.omega.-dicarboxylic
acid (12 mmol) was dissolved in dry THF, to that 1.2 equivalent of
DCC and catalytic amounts of DMAP was added at room temperature.
After stirring for 1 hr, 4-Hydroxyacetanilide (1.36 g, 10 mmol) was
added in one lot and stirring continued for 24 hrs. After
completion of the reaction, mixture was filtered through cindered
flask to remove DCU, washed with THF. After removal of THF,
slightly acidic water was added, and thrice extracted with
chloroform; organic layer was dried over anhydrous
Na.sub.2SO.sub.4. After evaporating the solvent, the obtained crude
products were purified by silica gel column chromatography using
methanol:chloroform (1:9) as eluent, afforded pure products as
white solid. Yields of the reactions were .about.50%.
[0159] Apn-7-COOH. .sup.1H-NMR, (CDCl.sub.3, 300 MHz) .delta. 7.48
(d, 2H), 7.01 (d, 2H), 2.54 (broad s, 1H), 2.35 (m, 4H), 2.15 (s,
3H), 1.70 (m, 4H), 1.5-1.2 (m, 4H).
[0160] Apn-13-COOH. .sup.1H-NMR, (CDCl.sub.3, 300 MHz) .delta. 7.50
(d, 2H), 7.05 (d, 2H), 2.57 (broad s, 1H), 2.38 (m, 4H), 2.2 (s,
3H), 1.77 (m, 4H), 1.5-1.2 (m, 16H).
[0161] General procedure for synthesis of Apn-8-Apn and Apn-14-Apn.
In a round bottom flask, .alpha.,.omega.-dicarboxylic acid (5 mmol)
was dissolved in dry THF, to that 2.5 equivalent of DCC and
catalytic amounts of DMAP was added at room temperature. After
stirring for 1 hr, 4-Hydroxyacetanilide (1.36 g, 10 mmol) was added
in one lot and stirring continued for 72 hrs. After completion of
the reaction, mixture was filtered through cindered flask to remove
DCU, washed with THF. After removal of THF, slightly acidic water
was added, and thrice extracted with chloroform; organic layer was
dried over anhydrous Na.sub.2SO.sub.4. After evaporating the
solvent, the obtained crude products were purified by silica gel
column chromatography using methanol:chloroform (0.5:9) as eluent,
afforded pure products as white solid. Yields of the reactions were
between 60-70%.
[0162] Apn-8-Apn. .sup.1H-NMR, (CDCl.sub.3, 300 MHz) .delta. 7.6
(d, 4H), 7.2 (d, 4H), 2.31 (broad s, 2H), 2.39 (m, 4H), 2.2 (s,
6H), 1.75 (m, 4H), 1.5-1.1 (m, 4H).
[0163] Apn-14-Apn. .sup.1H-NMR, (CDCl.sub.3, 300 MHz) .delta. 7.55
(d, 4H), 7.02 (d, 4H), 2.55 (broad s, 2H), 2.41 (m, 4H), 2.18 (s,
6H), 1.75 (m, 4H), 1.6-1.1 (m, 16H). Theoretical calculation
methods.
[0164] The geometry of acetaminophen and its derivatives was fully
optimized via ab initio calculations by using the restricted
Hartree-Fock theory with 6-31 G* basis set using GAUSSIAN 03
(Gaussian 03, Revision B.03, Frisch, M. J.; Trucks, G. W.;
Schlegel, H. B.; Scuseria, G. E.; Robb, M. A.; Cheeseman, J. R.;
Montgomery, Jr., J. A.; Vreven, T.; Kudin, K. N.; Burant, J. C.;
Millam, J. M.; Iyengar, S. S.; Tomasi, J.; Barone, V.; Mennucci,
B.; Cossi, M.; Scalmani, G.; Rega, N.; Petersson, G. A.; Nakatsuji,
H.; Hada, M.; Ehara, M.; Toyota, K.; Fukuda, R.; Hasegawa, J.;
Ishida, M.; Nakajima, T.; Honda, Y.; Kitao, O.; Nakai, H.; Klene,
M.; Li, X.; Knox, J. E.; Hratchian, H. P.; Cross, J. B.; Adamo, C.;
Jaramillo, J.; Gomperts, R.; Stratmann, R. E.; Yazyev, O.; Austin,
A. J.; Cammi, R.; Pomelli, C.; Ochterski, J. W.; Ayala, P. Y.;
Morokuma, K.; Voth, G. A.; Salvador, P.; Dannenberg, J. J.;
Zakrzewski, V. G.; Dapprich, S.; Daniels, A. D.; Strain, M. C.;
Farkas, O.; Malick, D. K.; Rabuck, A. D.; Raghavachari, K.;
Foresman, J. B.; Ortiz, J. V.; Cui, Q.; Baboul, A. G.; Clifford,
S.; Cioslowski, J.; Stefanov, B. B.; Liu, G.; Liashenko, A.;
Piskorz, P.; Komaromi, I.; Martin, R. L.; Fox, D. J.; Keith, T.;
Al-Laham, M. A.; Peng, C. Y.; Nanayakkara, A.; Challacombe, M.;
Gill, P. M. W.; Johnson, B.; Chen, W.; Wong, M. W.; Gonzalez, C.;
Pople, J. A. Gaussian, Inc., Pittsburgh Pa., 2003). Calculated
Cartesian coordinates are shown in Tables 4-6.
TABLE-US-00004 TABLE 4 Calculated Cartesian coordinates for Apn 6
2.605874 -0.218794 -0.000932 8 2.520379 -1.413547 -0.000769 7
1.539481 0.628244 -0.001902 1 1.753139 1.599461 -0.001818 6
3.945796 0.490784 0.002427 1 4.042112 1.138871 -0.862985 1 4.054076
1.100442 0.894075 1 4.729462 -0.251314 -0.018479 6 0.158994
0.325834 -0.001042 6 -0.729155 1.389986 -0.000759 6 -0.352592
-0.970403 -0.000591 6 -2.098728 1.181397 -0.000004 1 -0.357860
2.400944 -0.001129 6 -1.718751 -1.175710 0.000181 1 0.312612
-1.806716 -0.000836 6 -2.600567 -0.106508 0.000481 1 -2.765416
2.027266 0.000195 1 -2.114536 -2.174314 0.000542 8 -3.928022
-0.380401 0.001207 1 -4.434041 0.419708 0.001681
TABLE-US-00005 TABLE 5 Calculated Cartesian coordinates for
Apn-7-COOH 6 -7.766671 0.354033 0.000322 8 -7.658978 1.546282
0.000817 7 -6.713009 -0.512227 -0.001252 1 -6.942284 -1.480026
-0.001270 6 -9.117537 -0.333146 0.004351 1 -9.224976 -0.978287
-0.861896 1 -9.234274 -0.942362 0.895179 1 -9.889043 0.421603
-0.014532 6 -5.331831 -0.232714 -0.001190 6 -4.461451 -1.313119
-0.001768 6 -4.800070 1.053887 -0.000678 6 -3.085683 -1.147585
-0.001801 1 -4.854356 -2.315708 -0.002200 6 -3.429357 1.222264
-0.000705 1 -5.445675 1.905055 -0.000280 6 -2.571513 0.136034
-0.001241 1 -2.445310 -2.001611 -0.002217 1 -3.010634 2.211370
-0.000322 8 -1.233662 0.482398 -0.001340 6 -0.193431 -0.350064
-0.000546 8 -0.266735 -1.533968 0.000412 6 1.093206 0.440619
-0.000856 6 2.340339 -0.438076 -0.000039 1 1.072511 1.096034
0.865353 1 1.072817 1.094872 -0.867947 6 3.628351 0.386763
-0.000357 1 2.319015 -1.089047 0.868326 1 2.319334 -1.090209
-0.867542 6 4.888628 -0.479899 0.000448 1 3.642525 1.039355
-0.871461 1 3.642211 1.040515 0.869882 1 4.874503 -1.132519
0.871581 1 4.874817 -1.133678 -0.869821 6 6.176617 0.344724
0.000132 6 7.424223 -0.533270 0.000937 1 6.198357 0.996922 0.867519
1 6.198667 0.995771 -0.868112 1 7.446415 -1.187939 0.867940 1
7.446724 -1.189085 -0.865192 6 8.708587 0.254808 0.000646 8
8.803620 1.438826 -0.000143 8 9.775844 -0.542603 0.001347 1
10.554562 0.005512 0.001108
TABLE-US-00006 TABLE 6 Calculated Cartesian coordinates for
Apn-8-Apn 6 12.028399 0.305826 -0.008303 8 11.930570 1.498896
-0.020573 7 10.967686 -0.551783 0.002707 1 11.189210 -1.521350
0.011993 6 13.373520 -0.392437 -0.005656 1 13.477879 -1.024954
0.870242 1 13.482961 -1.016303 -0.887200 1 14.151294 0.356071
-0.000175 6 9.588754 -0.261421 0.000849 6 8.709964 -1.334786
0.012493 6 9.066943 1.029337 -0.011756 6 7.335402 -1.158540
0.011870 1 9.094885 -2.340422 0.022377 6 7.697685 1.208360
-0.012425 1 9.719161 1.875377 -0.020853 6 6.831254 0.128855
-0.000776 1 6.688591 -2.007651 0.020999 1 7.286632 2.200635
-0.022106 8 5.496198 0.485778 -0.003096 6 4.449457 -0.338640
0.004787 8 4.513806 -1.523015 0.015080 6 3.169013 0.461927
-0.001539 6 1.915004 -0.406909 0.006740 1 3.194192 1.108982
-0.873900 1 3.195118 1.124153 0.859337 6 0.633556 0.428063 0.000175
1 1.930537 -1.066143 -0.855485 1 1.931505 -1.051002 0.880316 6
-0.633604 -0.428466 0.008329 1 0.625260 1.088878 0.865162 1
0.624296 1.073720 -0.876173 1 -0.625325 -1.089247 -0.856682 1
-0.624327 -1.074156 0.884654 6 -1.915052 0.406506 0.001823 6
-3.169062 -0.462328 0.010038 1 -1.931566 1.050662 -0.871702 1
-1.930571 1.065680 0.864099 1 -3.195179 -1.124476 -0.850889 1
-3.194230 -1.109465 0.882348 6 -4.449506 0.338238 0.003820 8
-4.513879 1.522610 -0.006489 8 -5.496255 -0.486176 0.011252 6
-6.831262 -0.129096 0.008212 6 -7.335253 1.158291 0.026149 6
-7.697801 -1.208391 -0.011600 6 -8.709778 1.334770 0.023164 1
-6.688336 2.007243 0.041309 6 -9.067029 -1.029144 -0.014482 1
-7.286865 -2.200674 -0.024788 6 -9.588684 0.261621 0.003015 1
-9.094579 2.340404 0.036970 1 -9.719343 -1.875016 -0.030021 7
-10.967568 0.552203 0.001537 1 -11.188979 1.521742 0.015385 6
-12.028379 -0.305197 -0.014991 8 -11.930692 -1.498216 -0.032336 6
-13.373417 0.393196 -0.007972 1 -13.486330 0.998779 0.885788 1
-13.474197 1.043658 -0.871067 1 -14.151234 -0.354896 -0.032138
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Example 2: Self-Assembling Amphiphiles
[0208] A series of amphiphiles were designed for efficient
gelation. The amphiphile was synthesized in a single step that
avoids the harsh conditions and is beneficial when synthesizing on
an industrial scale. The use of expensive reagents was also
eliminated. Compound purification was relatively easy. After the
reaction, a simple filtration yielded a pure compound. Collectively
these features make the amphiphile attractive for use in various
biomedical applications.
[0209] This example focuses on a novel class of gelling agents or
thickeners that are based on readily available and economically
attractive starting materials. Gelling agents or thickeners capable
of gelling or thickening a wide variety of solvents were used,
making the gelling agents or thickeners suitable for employment in
various applications.
[0210] Exemplary compounds include those having the general
formula, RCONHC(CH.sub.2OH).sub.3, in which R is a saturated alkyl
chain having 9-15 C atoms or an unsaturated alkyl chain having 9-17
C atoms. N-tris(hydroxymethyl)methylamides of fatty acids, such as
lauric, caprylic, myristic, palmitic, and oleic acids, can be
prepared by reacting the fatty acids or their corresponding methyl
esters with tris(hydroxymethyl)methylamine in anhydrous
dimethylsulfoxide in the presence of potassium carbonate.
[0211] Results: These compounds have been found to produce a gel in
numerous solvents. Examples of suitable solvents include water,
salt water (such as a NaCl solution), different foreign bodies such
as TRISZMA buffers, and alkaline and acidic solutions. In addition
to water-based solutions, the compounds can gel various aromatic
and non-aromatic organic solvents, such as benzene, toluene,
hexane, and acetonitrile. See Table 7. All the gels obtained in
Table 7 were thermally reversible. Above their gelation
temperature, the gels dissolved in solvent, but could be returned
to their original gel state upon cooling.
[0212] Scanning electron microscopic studies. Molecular
self-aggregation features can be observed on an electron
microscope, since the initial stage of physical gelation is the
self-assembly of gelator monomers. FIGS. 11A-11B show the scanning
electron microscope (SEM) images of the organogels formed by the
amphiphiles in cyclohexane and water (FIGS. 11A and 11B,
respectively). Analysis of these aggregates showed that the
individual fibers are approximately 100-200 nm in wide and up to
several micrometers in length. These nanofibers were entangled and
formed a dense fibrous network, resulting in the immobilization of
the solvent.
[0213] The examples demonstrate the encapsulation of hydrophobic
and hydrophilic drugs in self-assembled hydrogels. Efficient
encapsulation of various drugs enables the utilization of these
gels for developing a wide range of therapeutics. A representative
self-assembly of amphiphiles to form gels, and encapsulation of
drugs in their nano- or micro structures is shown in FIG. 12.
TABLE-US-00007 TABLE 7 The gelation ability of polyol-12 and
polyol-16 in water and various organic solvents..sup.a solvent
po1yol-12 polyol-16 water G G NaCl (40 mM) G G acidic (pH <3) G
G basic (pH >12) G G TRIS (50 mM) G G cyclohexane G G benzene G
G toluene G G CCl.sub.4 G G CHCl.sub.3 P Sol CH.sub.3CN G G acetone
Sol Sol 1,4-dioxane Sol Sol DMSO Sol Sol .sup.aG = stable gel at
room temperature, P = precipitate, Sol = soluble.
References Considered for Example 2
[0214] 1. Whitesides, G. M. & Boncheva, M. (2002) Proc. Natl.
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van Esch, J. (2004) J. Controlled Release 97, 241-248. [0218] 5.
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128, 3038-3043. [0219] 6. van Bommel, K. J. C., Stuart, M. C. A.,
Feringa, B. L. & van Esch, J. (2005) Org. Biomol. Chem. 3,
2917-2920. [0220] 7. Miyata, T., Uragami, T. & Nakamae, K.
(2002) Adv. Drug Delivery Rev. 54, 79-98. [0221] 8. Menger, F. M.
& Caran, K. L. (2000) J. Am. Chem. Soc. 122, 11679-11691.
[0222] 9. Sreenivasachary, N. & Lehn, J.-M. (2005) Proc. Natl.
Acad. Sci. USA 102, 5938-5943. [0223] 10. Makarevic, J., Jokid, M.,
Percid, B., Tomisid, V., Krojid-Prodid, B. & Zinid, M. (2001)
Chem. Eur. J. 7, 3328-3341. [0224] 11. Oda, R., Huc, I. &
Candau, S. J. (1998) Angew. Chem. Int. Ed. 37, 2689-2691. [0225]
12. Estroff, L. A. & Hamilton, A. D. (2000) Angew. Chem. Int.
Ed. 39, 3447-3450. [0226] 13. Kobayashi, H., Friggeri, A., Koumoto,
K., Amaike, M., Shinkai, S. & Reinhoudt, D. N. (2002) Org.
Lett. 4, 1423-1426. [0227] 14. Luboradzki, R., Gronwald, O., Ikeda,
M., Shinkai, S. & Reinhoudt, D. N. (2000) Tetrahedron 56,
9595-9599. [0228] 15. Jung, J. H., John, G., Masuda, M., Yoshida,
K., Shinkai, S. & Shimizu, T. (2001) Langmuir 17, 7229-7232.
[0229] 16. Wang, G. & Hamilton, A. D. (2003) Chem. Commun.
310-311.
Example 3: Dye Encapsulated Self-Assembled Nanofibers: In Vitro and
In Vivo Analysis
[0230] In vitro: A model dye DiD was encapsulated within the
self-assembled nanofibers prepared from salicin prodrug of formula
(III). Salicin-deconate (4 wt/v %) was taken in glass vial and to
that dye and PBS were added. Homogenous mixture was formed upon
heating, subsequent cooling generated the gel. Fibers were isolated
by repetitive centrifugation and washing steps. Fibers were
incubated with synovial fluid at 37.degree. C. that was extracted
from human arthritic joints (inflamed). In the absence of synovial
fluid, fibers were stable and preserved the dye within assembled
fibers (FIG. 13A). The enzymes that were present in inflamed joints
degraded the fibers to release the encapsulated dye. Slow
degradation of fibers by enzymes over a period of 15 days was
observed (FIG. 13B). The data demonstrates that in the absence of
synovial fluid the fibers were intact and dye was confined to the
fibers, whereas in the presence of synovial fluid, the fibers
disassembled as observed by reduced number of fibers and by a
drastic drop in the dye intensity. An accumulation of background
fluorescence as the dye encapsulated fibers disassembled was also
observed. In the absence of synovial fluid the nanofibers were
stable (in PBS) for months.
[0231] In vivo: DiD encapsulated nanofiber gels (salicin-deconate
nanofibers, prepared as above), were directly injected into the
arthritis bearing mouse joints (ankles and knees) to examine the
retention of nanofibers within the joints. After 8 days, ankles and
knees were sectioned and imaged under fluorescence microscopy to
examine the presence of nanofibers. Results shown in FIG. 14A-14F
indicate that dye encapsulated nanofibers were localized within the
joints and a portion of the gel fibers originally injected can
remain stable for long periods of time.
[0232] In vivo: DiD encapsulated nanofiber gels were directly
injected into the arthritis bearing mouse joints (ankles and knees)
to examine the retention of nanofibers within the joints. After 8
days, ankles and knees were sectioned and imaged under fluorescence
microscopy to examine the presence of nanofibers. Results shown in
FIGS. 14A-14F indicate that dye encapsulated nanofibers were
localized within the joints and a portion of the gel fibers
originally injected can remain stable for long periods of time.
[0233] The present invention may be defined in any of the following
numbered paragraphs: [0234] 1. A drug-delivery composition,
comprising a drug-derived gelator capable of self assembly, wherein
the backbone of the gelator contains a drug and which drug can be
released upon a stimuli. [0235] 2. The composition of paragraph 1,
wherein the stimuli is enzyme-mediated degradation or hydrolytic
cleavage. [0236] 3. The composition of paragraph 1, wherein the
stimuli is of biological origin. [0237] 4. The composition of
paragraph 1, wherein the biological origin is inflammation. [0238]
5. The composition of paragraph 1, wherein the composition is a
hydrogel. [0239] 6. The composition of paragraph 1, wherein the
composition is self-assembling. [0240] 7. The composition of
paragraph 1, wherein the drug-derived gelator is a self-assembled
gelator. [0241] 8. The composition of paragraph 7, wherein the
gelator is a hydrogel or an organogelator. [0242] 9. The
composition of paragraph 1, wherein the backbone of the
drug-derived gelator contains the drug or a derivative thereof, a
hydrophobic moiety, and a degradable link between the drug and the
hydrophobic moiety. [0243] 10. The composition of paragraph 1,
wherein the drug is a hydrophobic drug or hydrophilic drug. [0244]
11. The composition of paragraph 1, wherein backbone of the
drug-derived gelator comprises at a hydrophobic drug and a
hydrophilic moiety. [0245] 12. The composition of paragraph 1,
wherein the drug-derived gelator comprises a hydrophilic drug and a
hydrophobic moiety. [0246] 13. The composition of paragraph 9,
wherein the drug-derived gelator is a drug derived, self-assembled
gelator. [0247] 14. The composition of paragraph 9, wherein the
hydrophobic moiety is a polymethylene chain or an aromatic group.
[0248] 15. The composition of paragraph 9, wherein the degradable
link is an ester, amide, anhydride, or carbamate linkage that is
cleavable upon contact with an enzyme and/or through hydrolysis.
[0249] 16. The composition of paragraph 1, wherein the stimuli is
temperature, pH, ultrasound, metal ions, light, electrical stimuli,
electromagnetic stimuli, or combinations thereof. [0250] 17. The
composition of paragraph 1, wherein the gelator is the
esterification reaction product of the drug and a C.sub.1-22
carboxylic acid. [0251] 18. The composition of paragraph 17,
wherein the carboxylic acid contains saturated or unsaturated
hydrocarbon chains. [0252] 19. The composition of paragraph 1,
wherein the drug contains a phenol ring and a terminal hydroxyl
group, and wherein the drug is capable of undergoing self assembly
upon esterification with a fatty acid or dicarboxylic acid. [0253]
20. The composition of paragraph 1, wherein the drug contains one
or more functional groups that promote self-assembly through pi-pi
stacking, hydrogen-bonding and/or van der Waals interactions.
[0254] 21. The composition of paragraph 13, wherein the moiety
conjugated to the drug contains one or more functional groups that
promote self-assembly through pi-pi stacking, hydrogen-bonding
and/or van der Waals interactions. [0255] 22. The composition of
paragraph 13, wherein both the drug and the moiety conjugated to
the drug contains one or more functional groups that promote
self-assembly through pi-pi stacking, hydrogen-bonding and van der
Waals interactions. [0256] 23. The composition of paragraph 20,
wherein the one or more functional groups that promote
self-assembly are selected from the group consisting of an aromatic
ring, amide, carbonyl group, urea group, polymethylene chain, and
combinations thereof. [0257] 24. The composition of paragraph 1,
wherein the drug contains one or more functional groups that
promote self-assembly upon esterification with a fatty acid or
dicarboxylic acid. [0258] 25. The composition of paragraph 7,
wherein the self-assembled gelator is a single-component gelator
that does not require any other complementary molecules or base
salts to induce self-assembly. [0259] 26. The composition of
paragraph 1, wherein the drug and/or gelator is biocompatible.
[0260] 27. The composition of paragraph 1, wherein the drug is
acetaminophen and the gelator is an acetaminophen-derived gelator.
[0261] 28. The composition of paragraph 27, wherein said
acetaminophen-derived gelator is selected from formula (I) or
formula (II):
##STR00011##
[0261] wherein n in each formula represents an integer from 1-22.
[0262] 29. The composition of paragraph 28, wherein the gelator is
represented by formula (I) and n is 1-12. [0263] 30. The
composition of paragraph 28, wherein the gelator is represented by
formula (II) and n is 1-12. [0264] 31. The composition of paragraph
1, wherein the drug is delivered in a sustained release. [0265] 32.
The composition of paragraph 7, wherein the composition further
comprises an agent encapsulated in the self-assembled gelator that
can be released with the drug upon enzyme-mediated degradation or
through hydrolytic cleavage of the gelator. [0266] 33. The
composition of paragraph 32, wherein the agent is a hydrophobic
drug encapsulated in the hydrophobic pockets of the self-assembled
gelator. [0267] 34. The composition of paragraph 32, wherein the
agent is a hydrophilic drug encapsulated in the self-assembled
gelator. [0268] 35. The composition of paragraph 32, where in the
agent is selected from the group consisting of organic dyes,
vitamins, derivatives of vitamins, peptides, proteins, enzymes,
RNAi, chemotherapeutics, anti-inflammatory drugs, and combinations
thereof. [0269] 36. A multi-drug delivery composition, comprising a
self-assembled gelator comprising a first drug and a second drug,
[0270] wherein the first drug is part of the backbone of the
gelator and the second drug is encapsulated in the gel of the
gelator, and [0271] wherein the first drug and the second drug can
be simultaneously released upon exposure to an enzyme or upon
hydrolytic cleavage of the gelator. [0272] 37. The composition of
paragraph 36, wherein the composition further comprises one or more
additional drugs in the backbone of the gelator and/or one or more
additional drugs encapsulated in the gel of the gelator. [0273] 38.
The composition of paragraph 36, wherein the first drug is an
analgesic or antipyretic drug and the second drug is an
anti-inflammatory agent. [0274] 39. The composition of paragraph
36, wherein the first drug is complementary to the second drug.
[0275] 40. The composition of paragraph 36, wherein both drugs are
delivered in a sustained release. [0276] 41. A drug-delivery
composition, comprising a drug and a self-assembled gelator having
an ester, amide, anhydride, or carbamate functionality and a
hydrophobic moiety in the backbone of the gelator, wherein the drug
is encapsulated in the gel of the gelator and can be released upon
enzyme-mediated degradation of the gelator using a suitable enzyme.
[0277] 42. The composition of paragraph 41, wherein the enzyme is
an amidase or metalloprotease enzyme. [0278] 43. The composition of
paragraph 41, wherein the gelator is a tris-based hydrogelator
having an amide functionality. [0279] 44. The composition of
paragraph m 41, wherein the gelator is the reaction product of
2-amino-2-hydroxymethylpropane-1,3-diol and a fatty acid or methyl
ester. [0280] 45. The composition of paragraph 41, wherein the
gelator is of formula (IV):
[0280] ##STR00012## [0281] wherein R is a saturated or unsaturated
C.sub.1-22 alkyl group, and R.sup.1, R.sup.2, and R.sup.3 each
independently represent a hydroxyalkyl group. [0282] 46. The
composition of paragraph 45, wherein R is a saturated C.sub.9-21
alkyl group or an unsaturated C.sub.9-21 alkyl group, and R.sup.1,
R.sup.2, and R.sup.3 each represent a hydroxymethyl group. [0283]
47. The composition of paragraph 46, wherein R is a saturated
C.sub.12-16 alkyl group [0284] 48. The composition of paragraph 41,
wherein the gelator is a single-component gelator that does not
require any other complementary molecules or base salts to induce
self-assembly. [0285] 49. The composition of paragraph 41, wherein
the drug is indomethacin. [0286] 50. The composition of paragraph
49, wherein indomethacin is connected to a peptide, fatty acid or
sugar moiety. [0287] 51. The composition of paragraph 49, wherein
the indomethacin is of formula (V):
[0287] ##STR00013## [0288] wherein R is peptide comprising 1-40
amino acids or R is saturated or unsaturated C.sub.1-C.sub.21 alkyl
group. [0289] 52. The composition of paragraph 41, wherein the drug
is ethambutol. [0290] 53. The composition of paragraph 52, wherein
ethambutol is connected to a peptide, fatty acid or sugar moiety.
[0291] 54. The composition of paragraph 52, wherein the ethambutol
is of formula (VI):
[0291] ##STR00014## [0292] wherein R is a peptide comprising 1-40
amino acids or R is saturated or unsaturated C.sub.1-21 alkyl
group. [0293] 55. A single-component drug-based gelator
composition, wherein the gelator comprises functional groups that
exist within the drug and/or within a modified moiety that is
conjugated to the drug, wherein the functional groups can interact
via pi-pi stacking, hydrophobic interactions, and/or hydrogen
bonding. [0294] 56. The composition of paragraph 55 wherein the
drug is released enzymatically and/or hydrolytically. [0295] 57. A
drug-delivery composition, comprising a prodrug-derived gelator
capable of self assembly, wherein the backbone of the gelator
contains a prodrug and which prodrug can be released upon
enzyme-mediated degradation or through hydrolytic cleavage of the
gelator. [0296] 58. The composition of paragraph 57, wherein the
prodrug-derived gelator is a self-assembled gelator. [0297] 59. The
composition of paragraph 58, wherein gelator is a hydrogelator or
an organogelator. [0298] 60. The composition of paragraph 58,
wherein the backbone of the prodrug-derived gelator contains the
prodrug or a derivative thereof, a hydrophobic moiety, and a
degradable link between the prodrug and the hydrophobic moiety.
[0299] 61. The composition of paragraph 58, wherein prodrug-derived
gelator contains the prodrug or a derivative thereof, a hydrophilic
moiety, and a degradable link between the prodrug and the
hydrophilic moiety. [0300] 62. The composition of paragraph 60 or
61, wherein the hydrophobic moiety is a polymethylene chain or an
aromatic ring. [0301] 63. The composition of paragraph 60 or 61,
wherein the degradable link is an ester, amide or carbamate linkage
that is cleavable upon contact with an enzyme and/or through
hydrolysis. [0302] 64. The composition of paragraph 58, wherein the
gelator is the esterification reaction product of the prodrug and a
C.sub.1-18 carboxylic acid. [0303] 65. The composition of paragraph
64, wherein the carboxylic acid consists of saturated or
unsaturated hydrocarbon chains. [0304] 66. The composition of
paragraph 58, wherein the prodrug contains polyhydroxyl groups, and
wherein the prodrug is capable of undergoing self assembly upon
esterification with a carboxylic acid. [0305] 67. The composition
of paragraph 58, wherein the prodrug is salicin and the gelator is
a salicin-derived gelator. [0306] 68. The composition of paragraph
67, wherein said prodrug-derived gelator is of formula
[0306] ##STR00015## [0307] wherein R is a saturated or unsaturated
C.sub.1-21 alkyl group. [0308] 69. The composition of paragraph 58,
wherein the composition further comprises an agent encapsulated in
the self-assembled gelator that can be released with the prodrug
upon enzyme-mediated degradation or through hydrolytic cleavage of
the gelator. [0309] 70. The composition of paragraph 1, wherein the
drug-derived gelator is a prodrug. [0310] 71. The composition of
paragraph 70, wherein the prodrugs are not covalently linked to
each other. [0311] 72. The composition of paragraph 1, wherein the
composition comprises a first drug-derived gelator and a second
drug derived gelator, wherein the first and second drug derived
gelators are different from each other. [0312] 73. The composition
of paragraph 72, wherein the first drug-derived gelator
non-covalently binds the second drug-derived gelator. [0313] 74.
The composition of any one of paragraph 1, wherein the composition
is a nano-particle or micro-particle. [0314] 75. The composition of
any one of paragraph 1-74, wherein the composition is lyophilized.
[0315] 76. The composition of any one of paragraphs 1-74, wherein
the drug from the drug-derived gelator is in biologically active
form upon release. [0316] 77. The composition of any one of
paragraphs 1-74, wherein amount of drug available for release is
defined by the composition of prodrug. [0317] 78. An injectable
self-titering drug delivery hydrogel composition comprising a
composition of any one of paragraphs 1-74, wherein amount of drug
released is proportional to amount of stimuli present. [0318] 79.
An injectable self-titering drug delivery hydrogel composition
comprising a composition of any one of paragraphs 1-74, wherein
rate of drug release is proportional to amount of stimuli present.
[0319] 80. A single-step method of making a low molecular weight,
amphiphilic prodrug, comprising the step of esterifying a compound
having a phenol ring and terminal hydroxyl group with a
dicarboxylic acid to form an amphiphilic prodrug containing an
ester functionality. [0320] 81. The method of paragraph 80, wherein
the esterification reaction takes place without the use of a
catalyst. [0321] 82. The method of paragraph 80, further comprising
the step of contacting the amphiphilic prodrug with a solvent to
induce self assembly and form a prodrug-based gelator. [0322] 83.
The method of paragraph 82, wherein the functional groups of the
gelator promote self assembly by facilitating hydrogen bonding,
intermolecular interactions through pi-pi stacking, and molecular
association through van der Waals interactions. [0323] 84. The
method of paragraph 82, wherein the solvent is water, an organic
solvent, or an oil. [0324] 85. The method of paragraph 82, wherein
the solvent is a water containing one or more salts. [0325] 86. The
method of paragraph 82, wherein the self assembly takes place in
the presence of one or more agents so that the agent or agents are
encapsulated in the gel of the gelator. [0326] 87. The method of
paragraph 86, wherein the agent is an enzyme. [0327] 88. The method
of paragraph 80, wherein the compound and the agent are both drugs.
[0328] 89. The method of paragraph 80, wherein the compound is
acetaminophen. [0329] 90. The method of paragraph 89, wherein the
amphiphilic prodrug is selected from formula (I) or formula
(II):
[0329] ##STR00016## [0330] wherein n in each formula represents an
integer from 1-22. [0331] 91. The method of paragraph 90, wherein
the gelator is represented by formula (I) and n is 1-12. [0332] 92.
The method of paragraph 90, wherein the gelator is represented by
formula (II) and n is 1-12. [0333] 93. The method of paragraph 80,
wherein the compound is indomethacin. [0334] 94. The method of
paragraph 93, wherein the indomethacin is of formula (V):
[0334] ##STR00017## [0335] wherein R is a peptide comprising 1-40
amino acids or R is saturated or unsaturated C.sub.1-21 alkyl
group. [0336] 95. The method of paragraph 1, wherein the compound
is ethambutol. [0337] 96. The method of paragraph 95, wherein the
ethambutol is of formula (VI):
[0337] ##STR00018## [0338] wherein R is a peptide comprising 1-40
amino acids or R is saturated or unsaturated C.sub.1-21 alkyl
group. [0339] 97. A single-step method of making a low molecular
weight, amphiphilic gelator, comprising the step of reacting a
tris-hydroxyl methyl amine with a fatty acid or methyl ester to
form an amphiphilic gelator having an amide functionality. [0340]
98. The method of paragraph 97, wherein the reaction takes place
without the use of a catalyst. [0341] 99. The method of paragraph
97, wherein the reaction takes place in dimethylsulfoxide and
K.sub.2C.sub.03 at about 20.degree. C. for about 24-48 hours.
[0342] 100. The method of paragraph 97, further comprising the step
of contacting the amphiphilic gelator with a solvent to induce self
assembly. [0343] 101. The method of paragraph 100, wherein the
solvent is water, an organic solvent, or an oil. [0344] 102. The
method of paragraph 100, wherein the solvent is water containing
one or more salts. [0345] 103. The method of paragraph 101, wherein
the organic solvent is selected from the group consisting of
hexane, heptane, benzene, toluene, mesitylene, ethylacetate,
chloroform, ethanol, propanol, butanol, tertiary butanol,
tetrahydrofuran, acetonitrile, and combinations thereof. [0346]
104. The method of paragraph 101, wherein the solvent oil is
selected from the group consisting of petrol, diesel, gasoline,
vegetable oil, cooking oil, polyunsaturated oils, and combinations
thereof. [0347] 105. The method of paragraph 97, wherein the self
assembly takes place in the presence of an agent so that the agent
is encapsulated in the gel of the gelator. [0348] 106. The method
of paragraph 97, wherein the amphiphilic gelator having an amide
functionality is of formula (IV):
[0348] ##STR00019## [0349] wherein R is a saturated or unsaturated
C.sub.1-22 alkyl group, and R.sup.1, R.sup.2, and R.sup.3 each
independently represent a hydroxyalkyl group. [0350] 107. The
method of paragraph 106, wherein R is a saturated C.sub.9-21 alkyl
group or an unsaturated C.sub.9-21 alkyl group, and R.sup.1,
R.sup.2, and R.sup.3 each represent a hydroxymethyl group. [0351]
108. A single-step method of making a low molecular weight,
amphiphilic prodrug, comprising the step of esterifying a compound
having a saccharide group, phenol ring and primary hydroxyl group
with a vinyl ester of carboxylic acid to form an amphiphilic
prodrug containing an ester functionality. [0352] 109. The method
of paragraph 108, wherein the esterification reaction takes place
in presence of a Lipase enzyme [0353] 110. The method of paragraph
109, wherein the Lipase enzyme is lipolase. [0354] 111. The method
of paragraph 108, wherein the reaction takes place at about
37-40.degree. C. for about 24-72 hours. [0355] 112. The method of
paragraph 111, wherein the reaction takes place under continuous
agitation. [0356] 113. The method of paragraph 108, wherein said
amphiphilic prodrug is of formula (III):
[0356] ##STR00020## [0357] wherein R is a saturated or unsaturated
C.sub.1-21 alkyl group. [0358] 114. A method of drug delivery,
comprising: [0359] administering to a patient in need thereof a
drug-derived or prodrug-derived gelator capable of self assembly,
wherein the backbone of the gelator contains a drug or prodrug, a
hydrophobic moiety, and a degradable link between the drug or
prodrug and hydrophobic moiety, and [0360] contacting the gelator
with an enzyme or water to cleave the degradable link and release
the drug or prodrug in the patient. [0361] 115. The method of
paragraph 114, wherein the drug-derived or prodrug-derived gelator
is a self-assembled gelator. [0362] 116. The method of paragraph
114, wherein the gelator is a hydrogelator or an organogelator.
[0363] 117. The method of paragraph 114, wherein the gelator is
contacted with a suitable enzyme, and the enzymatic cleavage takes
place under physiological conditions. [0364] 118. The method of
paragraph 114, wherein the gelator is contacted with water, and the
hydrolytic cleavage takes place under basic pH conditions. [0365]
119. The method of paragraph 114, wherein the drug or prodrug is
delivered in a sustained release without burst release. [0366] 120.
The method of paragraph 119, wherein the sustained release is
controlled by the concentration of the enzyme. [0367] 121. The
method of paragraph 114, wherein the degradable link is an ester,
amide, or carbamate bond. [0368] 122. The method of paragraph 114,
wherein the gelator is an acetaminophen-derived hydrogelator.
[0369] 123. The method of paragraph 122, wherein said
acetaminophen-derived gelator is selected from formula (I) or
formula (II):
[0369] ##STR00021## [0370] wherein n in each formula represents an
integer from 1-22. [0371] 124. The method of paragraph 114, wherein
the gelator is an indomethacin-derived hydrogelator. [0372] 125.
The method of paragraph 114, wherein the gelator is an
ethambutol-derived hydrogelator. [0373] 126. The method of
paragraph 114, wherein the gelator is a hydrogelator. [0374] 127.
The method of paragraph 123, wherein the gelator is represented by
formula (I) and n is 1-12. [0375] 128. The method of paragraph 123,
wherein the gelator is represented by formula (II) and n is 1-12.
[0376] 129. The method of paragraph 114, wherein the contacting
step releases the drug or prodrug and a fatty acid. [0377] 130. The
method of paragraph 115, wherein the gelator comprises a second
drug encapsulated in the self-assembled gelator. [0378] 131. The
method of paragraph 130, wherein the enzyme or water cleaves the
degradable bond in the gelator to release the encapsulated drug,
the drug or prodrug from the backbone, and a fatty acid. [0379]
132. The method of paragraph 130, wherein the encapsulated drug and
the drug or prodrug from the backbone are released simultaneously.
[0380] 133. The method of paragraph 114, wherein the gelator is a
salicin-derived gelator. [0381] 134. The method of paragraph 133,
wherein the gelator is of formula (III):
[0381] ##STR00022## [0382] wherein R is a saturated or unsaturated
C.sub.1-21 alkyl group. [0383] 135. A method of drug delivery,
comprising: [0384] administering to a patient in need thereof a
drug or prodrug encapsulated in a self-assembled gelator, wherein
the backbone of the gelator contains an amide functionality and a
hydrophobic moiety, and [0385] contacting the gelator with a
suitable enzyme or water to cleave the amide bond in the gelator
and release the drug or prodrug in the patient. [0386] 136. The
method of paragraph 135, wherein the gelator is contacted with a
suitable enzyme, and the enzymatic cleavage takes place under
physiological conditions [0387] 137. The method of paragraph 135,
wherein the gelator is contacted with water, and the hydrolytic
cleavage takes place under basic pH conditions. [0388] 138. The
method of paragraph 135, wherein the gelator is a tris-based
amphiphile. [0389] 139. The method of paragraph 138, wherein the
gelator is the reaction product of
2-amino-2-hydroxymethyl-propane-1,3-diol and a fatty acid or methyl
ester. [0390] 140. The method of paragraph 138, wherein the gelator
is of formula (IV):
[0390] ##STR00023## [0391] wherein R is a saturated or unsaturated
C.sub.1-22 alkyl group, and R.sup.1, R.sup.2, and R.sup.3 each
independently represent a hydroxyalkyl group. [0392] 141. The
method of paragraph 140, wherein R is a saturated C.sub.9-21 alkyl
group or an unsaturated C.sub.9-21 alkyl group, and R.sup.1,
R.sup.2, and R.sup.3 each represent a hydroxymethyl group. [0393]
142. The method of paragraph 135, wherein R is a saturated
C.sub.12-16 alkyl group [0394] 143. The method of paragraph 135,
wherein the contacting step releases the encapsulated drug or
prodrug and a fatty acid. [0395] 144. The method of paragraph 135,
wherein the gelator comprises a second drug encapsulated in the
self-assembled gelator. [0396] 145. The method of paragraph 144,
wherein the contacting step releases the encapsulated drug, the
derived drug or prodrug, and a fatty acid. [0397] 146. The method
of paragraph 144, wherein the encapsulated drug or prodrug and
derived drug are released simultaneously. [0398] 147. The method of
paragraph 136, wherein the enzyme is an external enzyme. [0399]
148. The method of paragraph 136, wherein the enzyme is
encapsulated in the gelator. [0400] 149. A method of drug delivery,
comprising: [0401] administering to a patient in need thereof a
drug or prodrug encapsulated in a self-assembled gelator, wherein
the backbone of the gelator contains a hydrophobic moiety, and a
degradable link between the drug or prodrug and hydrophobic moiety,
and [0402] contacting the gelator with a suitable enzyme cleave the
degradable link in the gelator and release the drug or prodrug in
the patient, wherein the cleavage takes place under physiological
conditions. [0403] 150. The method of paragraph 114, wherein the
drug is delivered via injection or implantation. [0404] 151. The
method of paragraph 150, wherein the gelator self assembles in
situ. [0405] 152. The method of paragraph 114, wherein the patient
is suffering from an arthritic ailment and at least one of the
drugs in the composition is an anti-inflammatory drug. [0406] 153.
The method of paragraph 119, wherein the sustained release is an
on-demand release. [0407] 154. A compound of formula (I):
[0407] ##STR00024## [0408] wherein n represents an integer from
1-22. [0409] 155. The compound of paragraph 154, wherein n ranges
from 1-12. [0410] 156. The compound of paragraph 155, wherein n
ranges from 6-12. [0411] 157. The compound of formula (II):
[0411] ##STR00025## [0412] wherein n represents an integer from
1-22. [0413] 158. The compound of paragraph 157, wherein n ranges
from 1-12. [0414] 159. The compound of paragraph 158, wherein n
ranges from 6-12. [0415] 160. The compound of formula (III):
[0415] ##STR00026## [0416] wherein R is a saturated or unsaturated
C.sub.1-21 alkyl group. [0417] 161. The compound of formula
(V):
[0417] ##STR00027## [0418] wherein R is a peptide comprising 1-40
amino acids or R is saturated or unsaturated C.sub.1-21 alkyl
group. [0419] 162. The compound of paragraph 161, wherein n ranges
from 1-12. [0420] 163. The compound of paragraph 161, wherein n
ranges from 6-12. [0421] 164. The compound of formula (VI):
[0421] ##STR00028## [0422] wherein R is a peptide comprising 1-40
amino acids or R is saturated or unsaturated C.sub.1-21 alkyl
group. [0423] 165. The compound of paragraph 164, wherein n ranges
from 1-12. [0424] 166. The compound of paragraph 165, wherein n
ranges from 6-12.
[0425] All patents and publications cited herein are hereby
incorporated by reference.
* * * * *